Science.gov

Sample records for agrobacterium-mediated stable transformation

  1. Development of an Agrobacterium-Mediated Stable Transformation Method for the Sensitive Plant Mimosa pudica

    PubMed Central

    Mano, Hiroaki; Fujii, Tomomi; Sumikawa, Naomi; Hiwatashi, Yuji; Hasebe, Mitsuyasu

    2014-01-01

    The sensitive plant Mimosa pudica has long attracted the interest of researchers due to its spectacular leaf movements in response to touch or other external stimuli. Although various aspects of this seismonastic movement have been elucidated by histological, physiological, biochemical, and behavioral approaches, the lack of reverse genetic tools has hampered the investigation of molecular mechanisms involved in these processes. To overcome this obstacle, we developed an efficient genetic transformation method for M. pudica mediated by Agrobacterium tumefaciens (Agrobacterium). We found that the cotyledonary node explant is suitable for Agrobacterium-mediated transformation because of its high frequency of shoot formation, which was most efficiently induced on medium containing 0.5 µg/ml of a synthetic cytokinin, 6-benzylaminopurine (BAP). Transformation efficiency of cotyledonary node cells was improved from almost 0 to 30.8 positive signals arising from the intron-sGFP reporter gene by using Agrobacterium carrying a super-binary vector pSB111 and stabilizing the pH of the co-cultivation medium with 2-(N-morpholino)ethanesulfonic acid (MES) buffer. Furthermore, treatment of the explants with the detergent Silwet L-77 prior to co-cultivation led to a two-fold increase in the number of transformed shoot buds. Rooting of the regenerated shoots was efficiently induced by cultivation on irrigated vermiculite. The entire procedure for generating transgenic plants achieved a transformation frequency of 18.8%, which is comparable to frequencies obtained for other recalcitrant legumes, such as soybean (Glycine max) and pea (Pisum sativum). The transgene was stably integrated into the host genome and was inherited across generations, without affecting the seismonastic or nyctinastic movements of the plants. This transformation method thus provides an effective genetic tool for studying genes involved in M. pudica movements. PMID:24533121

  2. Development of an Agrobacterium-mediated stable transformation method for the sensitive plant Mimosa pudica.

    PubMed

    Mano, Hiroaki; Fujii, Tomomi; Sumikawa, Naomi; Hiwatashi, Yuji; Hasebe, Mitsuyasu

    2014-01-01

    The sensitive plant Mimosa pudica has long attracted the interest of researchers due to its spectacular leaf movements in response to touch or other external stimuli. Although various aspects of this seismonastic movement have been elucidated by histological, physiological, biochemical, and behavioral approaches, the lack of reverse genetic tools has hampered the investigation of molecular mechanisms involved in these processes. To overcome this obstacle, we developed an efficient genetic transformation method for M. pudica mediated by Agrobacterium tumefaciens (Agrobacterium). We found that the cotyledonary node explant is suitable for Agrobacterium-mediated transformation because of its high frequency of shoot formation, which was most efficiently induced on medium containing 0.5 µg/ml of a synthetic cytokinin, 6-benzylaminopurine (BAP). Transformation efficiency of cotyledonary node cells was improved from almost 0 to 30.8 positive signals arising from the intron-sGFP reporter gene by using Agrobacterium carrying a super-binary vector pSB111 and stabilizing the pH of the co-cultivation medium with 2-(N-morpholino)ethanesulfonic acid (MES) buffer. Furthermore, treatment of the explants with the detergent Silwet L-77 prior to co-cultivation led to a two-fold increase in the number of transformed shoot buds. Rooting of the regenerated shoots was efficiently induced by cultivation on irrigated vermiculite. The entire procedure for generating transgenic plants achieved a transformation frequency of 18.8%, which is comparable to frequencies obtained for other recalcitrant legumes, such as soybean (Glycine max) and pea (Pisum sativum). The transgene was stably integrated into the host genome and was inherited across generations, without affecting the seismonastic or nyctinastic movements of the plants. This transformation method thus provides an effective genetic tool for studying genes involved in M. pudica movements. PMID:24533121

  3. Agrobacterium-Mediated Stable Genetic Transformation of Populus angustifolia and Populus balsamifera.

    PubMed

    Maheshwari, Priti; Kovalchuk, Igor

    2016-01-01

    The present study demonstrates Agrobacterium tumefaciens-mediated stable genetic transformation of two species of poplar - Populus angustifolia and Populus balsamifera. The binary vector pCAMBIA-Npro-long-Luc containing the luciferase reporter gene was used to transform stem internode and axillary bud explants. Putative transformants were regenerated on selection-free medium using our previously established in vitro regeneration method. Explant type, genotype, effect of pre-culture, Agrobacterium concentration, a time period of infection and varying periods of co-culture with bacteria were tested for the transformation frequency. The highest frequency of transformation was obtained with stem internode explants pre-cultured for 2 days, infected with Agrobacterium culture at the concentration of OD600 = 0.5 for 10 min and co-cultivated with Agrobacterium for 48 h. Out of the two genotypes tested, P. balsamifera exhibited a higher transformation rate in comparison to P. angustifolia. The primary transformants that exhibited luciferase activity in a bioluminescence assay under the CCD camera when subjected to polymerase chain reaction and Southern blot analysis revealed a stable single-copy integration of luc in their genomes. The reported protocol is highly reproducible and can be applied to other species of poplar; it will also be useful for future genetic engineering of one of the most important families of woody plants for sustainable development. PMID:27014319

  4. Agrobacterium-Mediated Stable Genetic Transformation of Populus angustifolia and Populus balsamifera

    PubMed Central

    Maheshwari, Priti; Kovalchuk, Igor

    2016-01-01

    The present study demonstrates Agrobacterium tumefaciens-mediated stable genetic transformation of two species of poplar – Populus angustifolia and Populus balsamifera. The binary vector pCAMBIA-Npro-long-Luc containing the luciferase reporter gene was used to transform stem internode and axillary bud explants. Putative transformants were regenerated on selection-free medium using our previously established in vitro regeneration method. Explant type, genotype, effect of pre-culture, Agrobacterium concentration, a time period of infection and varying periods of co-culture with bacteria were tested for the transformation frequency. The highest frequency of transformation was obtained with stem internode explants pre-cultured for 2 days, infected with Agrobacterium culture at the concentration of OD600 = 0.5 for 10 min and co-cultivated with Agrobacterium for 48 h. Out of the two genotypes tested, P. balsamifera exhibited a higher transformation rate in comparison to P. angustifolia. The primary transformants that exhibited luciferase activity in a bioluminescence assay under the CCD camera when subjected to polymerase chain reaction and Southern blot analysis revealed a stable single-copy integration of luc in their genomes. The reported protocol is highly reproducible and can be applied to other species of poplar; it will also be useful for future genetic engineering of one of the most important families of woody plants for sustainable development. PMID:27014319

  5. Agrobacterium-mediated transformation of Fusarium proliferatum.

    PubMed

    Bernardi-Wenzel, J; Quecine, M C; Azevedo, J L; Pamphile, J A

    2016-01-01

    Fusarium proliferatum is an important pathogen that is associated with plant diseases and primarily affects aerial plant parts by producing different mycotoxins, which are toxic to humans and animals. Within the last decade, this fungus has also been described as one of the causes of red root rot or sudden death syndrome in soybean, which causes extensive damage to this crop. This study describes the Agrobacterium tumefaciens-mediated transformation of F. proliferatum as a tool for the disruption of pathogenicity genes. The genetic transformation was performed using two binary vectors (pCAMDsRed and pFAT-GFP) containing the hph (hygromycin B resistance) gene as a selection marker and red and green fluorescence, respectively. The presence of acetosyringone and the use of filter paper or nitrocellulose membrane were evaluated for their effect on the transformation efficiency. A mean processing rate of 94% was obtained with 96 h of co-cultivation only in the presence of acetosyringone and the use of filter paper or nitrocellulose membrane did not affect the transformation process. Hygromycin B resistance and the presence of the hph gene were confirmed by PCR, and fluorescence due to the expression of GFP and DsRed protein was monitored in the transformants. A high rate of mitotic stability (95%) was observed. The efficiency of Agrobacterium-mediated transformation of F. proliferatum allows the technique to be used for random insertional mutagenesis studies and to analyze fungal genes involved in the infection process. PMID:27323127

  6. Agrobacterium-mediated transformation of three freshwater microalgal strains.

    PubMed

    Sanitha, Mary; Radha, Sudhakar; Fatima, Anwar Aliya; Devi, Selvaraju Gayathri; Ramya, Mohandass

    2014-01-01

    Microalgal transformation has gained interest in recent years. Agrobacterium-mediated transformation remains as the most efficient method for the development of transgenic plants and microalgae due to its wide host range, inexpensive procedure and transfer of large segments of DNA. In the present study, three different microalgal species were isolated from freshwater environment and identified based on the morphological characteristics and ITS-2 region amplification. Agrobacterium-mediated transformation was successful for the isolates Chlorella sp., Ankistrodesmus sp and Scenedesmus bajacalifornicus. Gene integration and expression was confirmed by PCR amplification of hptII and GUS histochemical assay. A. tumifaciens contamination was checked by amplification of npt II gene (kanamycin resistant) which lies outside the T-border. Based on GUS assay, transformation efficiencies were found to be 12.25% for Chlorella sp. 2.96% for Scenedesmus bajacalifornicus and 3.5% for Ankistrodesmus sp. PMID:25804057

  7. Agrobacterium-mediated transformation of maize (Zea mays) immature embryos.

    PubMed

    Lee, Hyeyoung; Zhang, Zhanyuan J

    2014-01-01

    Agrobacterium tumefaciens-mediated transformation is one of the most efficient and simple gene delivery systems for genetic improvement and biology studies in maize. This system has become more widely used by both public and private laboratories. However, transformation efficiencies vary greatly from laboratory to laboratory for the same genotype. Here, we illustrate our advanced Agrobacterium-mediated transformation method in Hi-II maize using simple binary vectors. The protocol utilizes immature embryos as starting explants and the bar gene as a selectable marker coupled with bialaphos as a selective agent. The protocol offers efficient transformation results with high reproducibility, provided that some experimental conditions are well controlled. This transformation method, with minor modifications, can be also employed to transform certain maize inbreds. PMID:24243211

  8. Is VIP1 important for Agrobacterium-mediated transformation?

    PubMed

    Shi, Yong; Lee, Lan-Ying; Gelvin, Stanton B

    2014-09-01

    Agrobacterium genetically transforms plants by transferring and integrating T-(transferred) DNA into the host genome. This process requires both Agrobacterium and host proteins. VirE2 interacting protein 1 (VIP1), an Arabidopsis bZIP protein, has been suggested to mediate transformation through interaction with and targeting of VirE2 to nuclei. We examined the susceptibility of Arabidopsis vip1 mutant and VIP1 overexpressing plants to transformation by numerous Agrobacterium strains. In no instance could we detect altered transformation susceptibility. We also used confocal microscopy to examine the subcellular localization of Venus-tagged VirE2 or Venus-tagged VIP1, in the presence or absence of the other untagged protein, in different plant cell systems. We found that VIP1-Venus localized in both the cytoplasm and the nucleus of Arabidopsis roots, agroinfiltrated Nicotiana benthamiana leaves, Arabidopsis mesophyll protoplasts and tobacco BY-2 protoplasts, regardless of whether VirE2 was co-expressed. VirE2 localized exclusively to the cytoplasm of tobacco and Arabidopsis protoplasts, whether in the absence or presence of VIP1 overexpression. In transgenic Arabidopsis plants and agroinfiltrated N. benthamina leaves we could occasionally detect small aggregates of the Venus signal in nuclei, but these were likely to be imagining artifacts. The vast majority of VirE2 remained in the cytoplasm. We conclude that VIP1 is not important for Agrobacterium-mediated transformation or VirE2 subcellular localization. PMID:24953893

  9. Aboveground insect infestation attenuates belowground Agrobacterium-mediated genetic transformation.

    PubMed

    Song, Geun Cheol; Lee, Soohyun; Hong, Jaehwa; Choi, Hye Kyung; Hong, Gun Hyong; Bae, Dong-Won; Mysore, Kirankumar S; Park, Yong-Soon; Ryu, Choong-Min

    2015-07-01

    Agrobacterium tumefaciens causes crown gall disease. Although Agrobacterium can be popularly used for genetic engineering, the influence of aboveground insect infestation on Agrobacterium induced gall formation has not been investigated. Nicotiana benthamiana leaves were exposed to a sucking insect (whitefly) infestation and benzothiadiazole (BTH) for 7 d, and these exposed plants were inoculated with a tumorigenic Agrobacterium strain. We evaluated, both in planta and in vitro, how whitefly infestation affects crown gall disease. Whitefly-infested plants exhibited at least a two-fold reduction in gall formation on both stem and crown root. Silencing of isochorismate synthase 1 (ICS1), required for salicylic acid (SA) synthesis, compromised gall formation indicating an involvement of SA in whitefly-derived plant defence against Agrobacterium. Endogenous SA content was augmented in whitefly-infested plants upon Agrobacterium inoculation. In addition, SA concentration was three times higher in root exudates from whitefly-infested plants. As a consequence, Agrobacterium-mediated transformation of roots of whitefly-infested plants was clearly inhibited when compared to control plants. These results suggest that aboveground whitefly infestation elicits systemic defence responses throughout the plant. Our findings provide new insights into insect-mediated leaf-root intra-communication and a framework to understand interactions between three organisms: whitefly, N. benthamiana and Agrobacterium. PMID:25676198

  10. Highly efficient Agrobacterium-mediated transformation of banana cv. Rasthali (AAB) via sonication and vacuum infiltration.

    PubMed

    Subramanyam, Kondeti; Subramanyam, Koona; Sailaja, K V; Srinivasulu, M; Lakshmidevi, K

    2011-03-01

    A reproducible and efficient transformation method was developed for the banana cv. Rasthali (AAB) via Agrobacterium-mediated genetic transformation of suckers. Three-month-old banana suckers were used as explant and three Agrobacterium tumefaciens strains (EHA105, EHA101, and LBA4404) harboring the binary vector pCAMBIA1301 were used in the co-cultivation. The banana suckers were sonicated and vacuum infiltered with each of the three A. tumefaciens strains and co-cultivated in the medium containing different concentrations of acetosyringone for 3 days. The transformed shoots were selected in 30 mg/l hygromycin-containing selection medium and rooted in rooting medium containing 1 mg/l IBA and 30 mg/l hygromycin. The presence and integration of the hpt II and gus genes into the banana genome were confirmed by GUS histochemical assay, polymerase chain reaction, and southern hybridization. Among the different combinations tested, high transformation efficiency (39.4 ± 0.5% GUS positive shoots) was obtained when suckers were sonicated and vacuum infiltered for 6 min with A. tumefaciens EHA105 in presence of 50 μM acetosyringone followed by co-cultivation in 50 μM acetosyringone-containing medium for 3 days. These results suggest that an efficient Agrobacterium-mediated transformation protocol for stable integration of foreign genes into banana has been developed and that this transformation system could be useful for future studies on transferring economically important genes into banana. PMID:21212957

  11. Agrobacterium-mediated transformation for the investigation of somatic recombination in the fungal pathogen Armillaria

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The honey fungus Armillaria mellea is a destructive soil-borne pathogen that affects over 300 plant species, and is of increasing interest due to its ability to decompose lignin. Here we report the transformation of this fungus. A range of techniques was evaluated, and Agrobacterium-mediated trans...

  12. A Fruiting Body Tissue Method for Efficient Agrobacterium-Mediated Transformation of Agaricus bisporus

    PubMed Central

    Chen, Xi; Stone, Michelle; Schlagnhaufer, Carl; Romaine, C. Peter

    2000-01-01

    We describe a modified Agrobacterium-mediated method for the efficient transformation of Agaricus bisporus. Salient features of this procedure include cocultivation of Agrobacterium and fruiting body gill tissue and use of a vector with a homologous promoter. This method offers new prospects for the genetic manipulation of this commercially important mushroom species. PMID:11010906

  13. Multiple host-cell recombination pathways act in Agrobacterium-mediated transformation of plant cells.

    PubMed

    Mestiri, Imen; Norre, Frédéric; Gallego, Maria E; White, Charles I

    2014-02-01

    Using floral-dip, tumorigenesis and root callus transformation assays of both germline and somatic cells, we present here results implicating the four major non-homologous and homologous recombination pathways in Agrobacterium-mediated transformation of Arabidopsis thaliana. All four single mutant lines showed similar mild reductions in transformability, but knocking out three of four pathways severely compromised Agrobacterium-mediated transformation. Although integration of T-DNA into the plant genome is severely compromised in the absence of known DNA double-strand break repair pathways, it does still occur, suggesting the existence of other pathways involved in T-DNA integration. Our results highlight the functional redundancy of the four major plant recombination pathways in transformation, and provide an explanation for the lack of strong effects observed in previous studies on the roles of plant recombination functions in transformation. PMID:24299074

  14. Highly efficient Agrobacterium-mediated transformation of Volvariella volvacea.

    PubMed

    Wang, Jie; Guo, Liqiong; Zhang, Kai; Wu, Qi; Lin, Junfang

    2008-11-01

    Agrobacterium tumefaciens-mediated transformation (ATMT) was successfully applied to the edible straw mushroom, Volvariella volvacea. Mycelium pellets were transformed to cold stress resistance using the afp gene as both a selective marker and a reporter gene, under the control of a heterologous Lentinula edodes gpd promoter. The efficiency of transformation is over 100 times higher than that previously reported in V. volvacea. Stable integration of the afp gene with 1-4 copy numbers was confirmed in all 10 randomly selected transgenic events by Southern blot analysis. The mitotic stability of the transformants was demonstrated after five successive transfers on PDA medium without selection pressure and the PCR analysis of basidiospores harvested from transformants. PMID:18434137

  15. A rapid, highly efficient and economical method of Agrobacterium-mediated in planta transient transformation in living onion epidermis.

    PubMed

    Xu, Kedong; Huang, Xiaohui; Wu, Manman; Wang, Yan; Chang, Yunxia; Liu, Kun; Zhang, Ju; Zhang, Yi; Zhang, Fuli; Yi, Liming; Li, Tingting; Wang, Ruiyue; Tan, Guangxuan; Li, Chengwei

    2014-01-01

    Transient transformation is simpler, more efficient and economical in analyzing protein subcellular localization than stable transformation. Fluorescent fusion proteins were often used in transient transformation to follow the in vivo behavior of proteins. Onion epidermis, which has large, living and transparent cells in a monolayer, is suitable to visualize fluorescent fusion proteins. The often used transient transformation methods included particle bombardment, protoplast transfection and Agrobacterium-mediated transformation. Particle bombardment in onion epidermis was successfully established, however, it was expensive, biolistic equipment dependent and with low transformation efficiency. We developed a highly efficient in planta transient transformation method in onion epidermis by using a special agroinfiltration method, which could be fulfilled within 5 days from the pretreatment of onion bulb to the best time-point for analyzing gene expression. The transformation conditions were optimized to achieve 43.87% transformation efficiency in living onion epidermis. The developed method has advantages in cost, time-consuming, equipment dependency and transformation efficiency in contrast with those methods of particle bombardment in onion epidermal cells, protoplast transfection and Agrobacterium-mediated transient transformation in leaf epidermal cells of other plants. It will facilitate the analysis of protein subcellular localization on a large scale. PMID:24416168

  16. Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation.

    PubMed

    Shcherbak, N; Kishchenko, O; Sakhno, L; Komarnytsky, I; Kuchuk, M

    2013-01-01

    Lox sites of the Cre/lox recombination system from bacteriophage P1 were analyzed for their ability to affect on transgene expression when inserted upstream from a gene coding sequence adjacent to the right border (RB) of T-DNA. Wild and mutated types of lox sites were tested for their effect upon bar gene expression in plants obtained via Agrobacterium-mediated and biolistic transformation methods. Lox-mediated expression of bar gene, recognized by resistance of transgenic plants to PPT, occurred only in plants obtained via Agrobacterium-mediated transformation. RT-PCR analysis confirms that PPT-resistant phenotype of transgenic plants obtained via Agrobacterium-mediated transformation was caused by activation of bar gene. The plasmid with promoterless gus gene together with the lox site adjacent to the RB was constructed and transferred to Nicotiana tabacum as well. Transgenic plants exhibited GUS activity and expression of gus gene was detected in plant leaves. Expression of bar gene from the vectors containing lox site near RB allowed recovery of numerous PPT-resistant transformants of such important crops as Beta vulgaris, Brassica napus, Lactuca sativa and Solanum tuberosum. Our results demonstrate that the lox site sequence adjacent to the RB can be used to control bar gene expression in transgenic plants. PMID:23821951

  17. Strategies to improve low copy transgenic events in Agrobacterium-mediated transformation of maize.

    PubMed

    Sivamani, Elumalai; Li, Xianggan; Nalapalli, Samson; Barron, Yoshimi; Prairie, Anna; Bradley, David; Doyle, Michele; Que, Qiudeng

    2015-12-01

    Transgenic plants containing low copy transgene insertion free of vector backbone are highly desired for many biotechnological applications. We have investigated two different strategies for increasing the percentage of low copy events in Agrobacterium-mediated transformation experiments in maize. One of the strategies is to use a binary vector with two separate T-DNAs, one T-DNA containing an intact E.coli manA gene encoding phosphomannose isomerase (PMI) as selectable marker gene cassette and another T-DNA containing an RNAi cassette of PMI sequences. By using this strategy, low copy transgenic events containing the transgenes were increased from 43 to 60 % in maize. An alternate strategy is using selectable marker gene cassettes containing regulatory or coding sequences derived from essential plant genes such as 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) or MADS box transcription factor. In this paper we demonstrate that higher percentage of low copy transgenic events can be obtained in Agrobacterium-mediated maize transformation experiments using both strategies. We propose that the above two strategies can be used independently or in combination to increase transgenic events that contain low copy transgene insertion in Agrobacterium-mediated transformation experiments. PMID:26338266

  18. A Novel Phenolic Compound, Chloroxynil, Improves Agrobacterium-Mediated Transient Transformation in Lotus japonicus

    PubMed Central

    Kimura, Mitsuhiro; Cutler, Sean; Isobe, Sachiko

    2015-01-01

    Agrobacterium-mediated transformation is a commonly used method for plant genetic engineering. However, the limitations of Agrobacterium host-plant interactions and the complexity of plant tissue culture often make the production of transgenic plants difficult. Transformation efficiency in many legume species, including soybean and the common bean, has been reported to be quite low. To improve the transformation procedure in legumes, we screened for chemicals that increase the transformation efficiency of Lotus japonicus, a model legume species. A Chemical library was screened and chemicals that increase in transient transformation efficiency of L. japonicus accession, Miyakojima MG-20 were identified. The transient transformation efficiency was quantified by reporter activity in which an intron-containing reporter gene produces the GUS protein only when the T-DNA is expressed in the plant nuclei. We identified a phenolic compound, chloroxynil, which increased the genetic transformation of L. japonicus by Agrobacterium tumefaciens strain EHA105. Characterization of the mode of chloroxynil action indicated that it enhanced Agrobacterium-mediated transformation through the activation of the Agrobacterium vir gene expression, similar to acetosyringone, a phenolic compound known to improve Agrobacterium-mediated transformation efficiency. Transient transformation efficiency of L. japonicus with 5 μM chloroxynil was 60- and 6- fold higher than that of the control and acetosyringone treatment, respectively. In addition, transgenic L. japonicus lines were successfully generated by 5 μM chloroxynil treatment.Furthermore, we show that chloroxynil improves L. japonicus transformation by Agrobacterium strain GV3101 and rice transformation. Our results demonstrate that chloroxynil significantly improves Agrobacterium tumefaciens-mediated transformation efficiency of various agriculturally important crops. PMID:26176780

  19. An embryogenic suspension cell culture system for Agrobacterium-mediated transformation of citrus.

    PubMed

    Dutt, M; Grosser, J W

    2010-11-01

    A method for the genetic transformation of several citrus cultivars is described, including cultivars observed to be recalcitrant to conventional epicotyl-mediated transformation. Embryogenic cell suspension cultures, established from unfertilized ovules were used as target tissues for Agrobacterium-mediated transformation. Several modifications were made to the culture environment to investigate factors required for efficient transfer of the T-DNA and the subsequent regeneration of transgenic citrus plants. It was determined that co-cultivation of citrus cells and Agrobacterium in EME medium supplemented with maltose (EME-M) and 100 μM acetosyringone for 5 days at 25°C was optimum for transformation of each of the citrus cultivars. Efficient selection was obtained and escapes were prevented when the antibiotic hygromycin B was used as a selection antibiotic following transformation with an Agrobacterium strain containing hptII in the T-DNA region. Transgenic embryo regeneration and development was enhanced in medium that contained a liquid overlay consisting of a 1:2 mixture of 0.6 M BH3 and 0.15 M EME-M media. PCR and Southern blot analyses confirmed the presence of the T-DNA and the stable integration into the genome of regenerated plants, while RT-PCR demonstrated variable amounts of RNA being transcribed in different transgenic lines. This protocol can create an avenue for insertion of useful traits into any polyembryonic citrus cultivar that can be established as embryogenic cell suspension cultures, including popular specialty mandarins and seedless cultivars. PMID:20711728

  20. Agrobacterium-mediated genetic transformation of Prunus salicina

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report Agrobacterium tumefaciens-mediated transformation from hypocotyls slices of two Prunus salicina varieties, 'Angeleno' and 'Larry Anne', using a modification of the technique previously described for P. domestica. Regeneration rates on thidiazuron (TDZ) and indole-3-butyric acid (IBA) supp...

  1. Genetic transformation of wheat via Agrobacterium-mediated DNA delivery.

    PubMed

    Sparks, Caroline A; Doherty, Angela; Jones, Huw D

    2014-01-01

    The method described involves an initial incubation of wheat immature embryos in a liquid culture of Agrobacterium tumefaciens. The Agrobacterium strain is engineered to contain a binary vector with a gene of interest and a selectable marker gene placed between the T-DNA borders; the T-DNA is the region transferred to the plant cells, thus harnessing the bacterium's natural ability to deliver specific DNA into host cells. Following the initial inoculation with the Agrobacterium, the embryos are co-cultivated for several days after which the Agrobacterium is selectively destroyed using an antibiotic. Tissue culture of the embryos on plant media with a correct balance of hormones allows embryogenic callus formation followed by regeneration of plantlets, and in the later stages of tissue culture a selectable marker (herbicide) is included to minimize the incidence of non-transformed plants. This protocol has been used successfully to generate transformed plants of a wide range of wheat varieties, both spring and winter bread wheats (T. aestivum L.) and durum wheats (T. turgidum L.). PMID:24243208

  2. Agrobacterium-mediated transformation of tomato with the ICE1 transcription factor gene.

    PubMed

    Juan, J X; Yu, X H; Jiang, X M; Gao, Z; Zhang, Y; Li, W; Duan, Y D; Yang, G

    2015-01-01

    ICE1 genes play a very important role in plants in cold conditions. To improve the cold resistance of tomato, the ICE1 gene of Arabidopsis thaliana was used to construct the plant expression vector p3301-ICE1, and was overexpressed in tomato through Agrobacterium-mediated transformation. Five strains of resistant plants were obtained. PCR and half-quantitative results showed that the ICE1 gene was transferred to tomato; three strains tested positive. After low-temperature stress treatment, praline content and peroxide and catalase activities in the transgenic tomato plants were higher compared with non-transgenic controls, while malondialdehyde content was clearly lower. PMID:25729995

  3. Agrobacterium-mediated transformation of friable embryogenic calli and regeneration of transgenic cassava.

    PubMed

    Bull, S E; Owiti, J A; Niklaus, M; Beeching, J R; Gruissem, W; Vanderschuren, H

    2009-01-01

    Agrobacterium-mediated transformation of friable embryogenic calli (FEC) is the most widely used method to generate transgenic cassava plants. However, this approach has proven to be time-consuming and can lead to changes in the morphology and quality of FEC, influencing regeneration capacity and plant health. Here we present a comprehensive, reliable and improved protocol, taking approximately 6 months, that optimizes Agrobacterium-mediated transformation of FEC from cassava model cultivar TMS60444. We cocultivate the FEC with Agrobacterium directly on the propagation medium and adopt the extensive use of plastic mesh for easy and frequent transfer of material to new media. This minimizes stress to the FEC cultures and permits a finely balanced control of nutrients, hormones and antibiotics. A stepwise increase in antibiotic concentration for selection is also used after cocultivation with Agrobacterium to mature the transformed FEC before regeneration. The detailed information given here for each step should enable successful implementation of this technology in other laboratories, including those being established in developing countries where cassava is a staple crop. PMID:20010938

  4. Agrobacterium-Mediated Transformation of the Recalcitrant Vanda Kasem's Delight Orchid with Higher Efficiency

    PubMed Central

    Gnasekaran, Pavallekoodi; James Antony, Jessica Jeyanthi; Uddain, Jasim; Subramaniam, Sreeramanan

    2014-01-01

    The presented study established Agrobacterium-mediated genetic transformation using protocorm-like bodies (PLBs) for the production of transgenic Vanda Kasem's Delight Tom Boykin (VKD) orchid. Several parameters such as PLB size, immersion period, level of wounding, Agrobacterium density, cocultivation period, and concentration of acetosyringone were tested and quantified using gusA gene expression to optimize the efficiency of Agrobacterium-mediated genetic transformation of VKD's PLBs. Based on the results, 3-4 mm PLBs wounded by scalpel and immersed for 30 minutes in Agrobacterium suspension of 0.8 unit at A600nm produced the highest GUS expression. Furthermore, cocultivating infected PLBs for 4 days in the dark on Vacin and Went cocultivation medium containing 200 𝜇M acetosyringone enhanced the GUS expression. PCR analysis of the putative transformants selected in the presence of 250 mg/L cefotaxime and 30 mg/L geneticin proved the presence of wheatwin1, wheatwin2, and nptII genes. PMID:24977213

  5. Agrobacterium-mediated transformation of two Serbian potato cultivars (Solanum tuberosum L. cv. Dragacevka and cv. Jelica)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An efficient protocol for Agrobacterium-mediated transformation of Serbian potato cultivars Dragacevka and Jelica, enabling the introduction of oryzacystatin genes OCI and OCII, was established. Starting with leaf explants a two-stage transformation protocol combining procedures of Webb and Wenzler...

  6. Mature seed-derived callus of the model indica rice variety Kasalath is highly competent in Agrobacterium-mediated transformation.

    PubMed

    Saika, Hiroaki; Toki, Seiichi

    2010-12-01

    We previously established an efficient Agrobacterium-mediated transformation system using primary calli derived from mature seeds of the model japonica rice variety Nipponbare. We expected that the shortened tissue culture period would reduce callus browning--a common problem with the indica transformation system during prolonged tissue culture in the undifferentiated state. In this study, we successfully applied our efficient transformation system to Kasalath--a model variety of indica rice. The Luc reporter system is sensitive enough to allow quantitative analysis of the competency of rice callus for Agrobacterium-mediated transformation. We unexpectedly discovered that primary callus of Kasalath exhibits a remarkably high competency for Agrobacterium-mediated transformation compared to Nipponbare. Southern blot analysis and Luc luminescence showed that independent transformation events in primary callus of Kasalath occurred successfully at ca. tenfold higher frequency than in Nipponbare, and single copy T-DNA integration was observed in ~40% of these events. We also compared the competency of secondary callus of Nipponbare and Kasalath and again found superior competency in Kasalath, although the identification and subsequent observation of independent transformation events in secondary callus is difficult due to the vigorous growth of both transformed and non-transformed cells. An efficient transformation system in Kasalath could facilitate the identification of QTL genes, since many QTL genes are analyzed in a Nipponbare × Kasalath genetic background. The higher transformation competency of Kasalath could be a useful trait in the establishment of highly efficient systems involving new transformation technologies such as gene targeting. PMID:20853107

  7. Agrobacterium-mediated transformation of Vitis Cv. Monastrell suspension-cultured cells: Determination of critical parameters.

    PubMed

    Chu, Mingyu; Quiñonero, Carmen; Akdemir, Hülya; Alburquerque, Nuria; Pedreño, María Ángeles; Burgos, Lorenzo

    2016-05-01

    Although some works have explored the transformation of differentiated, embryogenic suspension-cultured cells (SCC) to produce transgenic grapevine plants, to our knowledge this is one of the first reports on the efficient transformation of dedifferentiated Vitis vinifera cv Monastrell SCC. This protocol has been developed using the sonication-assisted Agrobacterium-mediated transformation (SAAT) method. A construct harboring the selectable nptII and the eyfp/IV2 marker genes was used in the study and transformation efficiencies reached over 50 independent transformed SCC per gram of infected cells. Best results were obtained when cells were infected at the exponential phase. A high density plating (500 mg/dish) gave significantly better results. As selective agent, kanamycin was inefficient for the selection of Monastrell transformed SCC since wild type cells were almost insensitive to this antibiotic whereas application of paromomycin resulted in very effective selection. Selected eyfp-expressing microcalli were grown until enough tissue was available to scale up a new transgenic SCC. These transgenic SCC lines were evaluated molecularly and phenotypically demonstrating the presence and integration of both transgenes, the absence of Agrobacterium contamination and the ability of the transformed SCC to grow in highly selective liquid medium. The methodology described here opens the possibility of improving the production of valuable metabolites. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:725-734, 2016. PMID:26871543

  8. Enhanced Agrobacterium-mediated transformation efficiencies in monocot cells is associated with attenuated defense responses.

    PubMed

    Zhang, Wan-Jun; Dewey, Ralph E; Boss, Wendy; Phillippy, Brian Q; Qu, Rongda

    2013-02-01

    Plant defense responses can lead to altered metabolism and even cell death at the sites of Agrobacterium infection, and thus lower transformation frequencies. In this report, we demonstrate that the utilization of culture conditions associated with an attenuation of defense responses in monocot plant cells led to highly improved Agrobacterium-mediated transformation efficiencies in perennial ryegrass (Lolium perenne L.). The removal of myo-inositol from the callus culture media in combination with a cold shock pretreatment and the addition of L-Gln prior to and during Agrobacterium-infection resulted in about 84 % of the treated calluses being stably transformed. The omission of myo-inositol from the callus culture media was associated with the failure of certain pathogenesis related genes to be induced after Agrobacterium infection. The addition of a cold shock and supplemental Gln appeared to have synergistic effects on infection and transformation efficiencies. Nearly 60 % of the stably transformed calluses regenerated into green plantlets. Calluses cultured on media lacking myo-inositol also displayed profound physiological and biochemical changes compared to ones cultured on standard growth media, such as reduced lignin within the cell walls, increased starch and inositol hexaphosphate accumulation, enhanced Agrobacterium binding to the cell surface, and less H(2)O(2) production after Agrobacterium infection. Furthermore, the cold treatment greatly reduced callus browning after infection. The simple modifications described in this report may have broad application for improving genetic transformation of recalcitrant monocot species. PMID:23242917

  9. AGROBACTERIUM-MEDIATED TRANSFORMATION IN THE GREEN ALGA HAEMATOCOCCUS PLUVIALIS (CHLOROPHYCEAE, VOLVOCALES)(1).

    PubMed

    Kathiresan, S; Chandrashekar, A; Ravishankar, G A; Sarada, R

    2009-06-01

    The first successful Agrobacterium-mediated transformation of the green alga Haematococcus pluvialis Flot. using the binary vectors hosting the genes coding for GUS (β-glucuronidase), GFP (green fluorescent protein), and hpt (hygromycin phosphotransferase) is reported here. Colonies resistant to hygromycin at 10 mg · L(-1) expressed β-glucuronidase. The greenish yellow fluorescence of GFP was observed when the hygromycin-resistant cells were viewed with a fluorescent microscope. PCR was used to successfully amplify fragments of the hpt (407 bp) and GUS (515 bp) genes from transformed cells, while Southern blots indicated the integration of the hygromycin gene into the genome of H. pluvialis. SEM indicated that the cell wall of H. pluvialis was altered on infection with Agrobacterium. The transformation achieved here by Agrobacterium does not need treatment with acetosyringone or the wounding of cells. A robust transformation method for this alga would pave the way for manipulation of many important pathways relevant to the food, pharmaceutical, and nutraceutical industries. PMID:27034041

  10. Origin of somatic embryos from repetitively embryogenic cultures of walnut (Juglans regia L.): Implications forAgrobacterium-mediated transformation.

    PubMed

    Polito, V S; McGranahan, G; Pinney, K; Leslie, C

    1989-04-01

    Early stages of somatic embryo development from embryogenic cultures ofJuglans regia (Persian or English walnut) are described. Histological examination reveals that secondary somatic embryos arise from cotyledons and hypocotyls of primary embryos cultured in the dark. The embryos originate by transverse to oblique divisions of surface cells. Single-cell origin of the secondary embryos confirms the potential of the repetitive embryogenesis system forAgrobacterium-mediated transformation and regeneration of non-chimeric, transgenic walnut plants. PMID:24233141

  11. Generation of transgenic plants of a potential oilseed crop Camelina sativa by Agrobacterium-mediated transformation.

    PubMed

    Lu, Chaofu; Kang, Jinling

    2008-02-01

    Camelina sativa is an alternative oilseed crop that can be used as a potential low-cost biofuel crop or a source of health promoting omega-3 fatty acids. Currently, the fatty acid composition of camelina does not uniquely fit any particular uses, thus limit its commercial value and large-scale production. In order to improve oil quality and other agronomic characters, we have developed an efficient and simple in planta method to generate transgenic camelina plants. The method included Agrobacterium-mediated inoculation of plants at early flowering stage along with a vacuum infiltration procedure. We used a fluorescent protein (DsRed) as a visual selection marker, which allowed us to conveniently screen mature transgenic seeds from a large number of untransformed seeds. Using this method, over 1% of transgenic seeds can be obtained. Genetic analysis revealed that most of transgenic plants contain a single copy of transgene. In addition, we also demonstrated that transgenic camelina seeds produced novel hydroxy fatty acids by transforming a castor fatty acid hydroxylase. In conclusion, our results provide a rapid means to genetically improve agronomic characters of camelina, including fatty acid profiles of its seed oils. Camelina may serve as a potential industrial crop to produce novel biotechnology products. PMID:17899095

  12. Agrobacterium-mediated transformation of durum wheat (Triticum turgidum L. var. durum cv Stewart) with improved efficiency

    PubMed Central

    He, Y.; Jones, H. D.; Chen, S.; Chen, X. M.; Wang, D. W.; Li, K. X.; Wang, D. S.; Xia, L. Q.

    2010-01-01

    An efficient Agrobacterium-mediated durum wheat transformation system has been developed for the production of 121 independent transgenic lines. This improved system used Agrobacterium strain AGL1 containing the superbinary pGreen/pSoup vector system and durum wheat cv Stewart as the recipient plant. Acetosyringone at 400 μM was added to both the inoculation and cultivation medium, and picloram at 10 mg l−1 and 2 mg l−1 was used in the cultivation and induction medium, respectively. Compared with 200 μM in the inoculation and cultivation media, the increased acetosyringone concentration led to significantly higher GUS (β-glucuronidase) transient expression and T-DNA delivery efficiency. However, no evident effects of acetosyringone concentration on regeneration frequency were observed. The higher acetosyringone concentration led to an improvement in average final transformation efficiency from 4.7% to 6.3%. Furthermore, the concentration of picloram in the co-cultivation medium had significant effects on callus induction and regeneration. Compared with 2 mg l−1 picloram in the co-cultivation medium, increasing the concentration to 10 mg l−1 picloram resulted in improved final transformation frequency from 2.8% to 6.3%, with the highest frequency of 12.3% reached in one particular experiment, although statistical analysis showed that this difference in final transformation efficiency had a low level of significance. Stable integration of foreign genes, their expression, and inheritance were confirmed by Southern blot analyses, GUS assay, and genetic analysis. Analysis of T1 progeny showed that, of the 31 transgenic lines randomly selected, nearly one-third had a segregation ratio of 3:1, while the remainder had ratios typical of two or three independently segregating loci. PMID:20202997

  13. High-efficiency Agrobacterium-mediated transformation of Norway spruce (Picea abies) and loblolly pine (Pinus taeda)

    NASA Technical Reports Server (NTRS)

    Wenck, A. R.; Quinn, M.; Whetten, R. W.; Pullman, G.; Sederoff, R.; Brown, C. S. (Principal Investigator)

    1999-01-01

    Agrobacterium-mediated gene transfer is the method of choice for many plant biotechnology laboratories; however, large-scale use of this organism in conifer transformation has been limited by difficult propagation of explant material, selection efficiencies and low transformation frequency. We have analyzed co-cultivation conditions and different disarmed strains of Agrobacterium to improve transformation. Additional copies of virulence genes were added to three common disarmed strains. These extra virulence genes included either a constitutively active virG or extra copies of virG and virB, both from pTiBo542. In experiments with Norway spruce, we increased transformation efficiencies 1000-fold from initial experiments where little or no transient expression was detected. Over 100 transformed lines expressing the marker gene beta-glucuronidase (GUS) were generated from rapidly dividing embryogenic suspension-cultured cells co-cultivated with Agrobacterium. GUS activity was used to monitor transient expression and to further test lines selected on kanamycin-containing medium. In loblolly pine, transient expression increased 10-fold utilizing modified Agrobacterium strains. Agrobacterium-mediated gene transfer is a useful technique for large-scale generation of transgenic Norway spruce and may prove useful for other conifer species.

  14. Agrobacterium-mediated genetic transformation using cotyledons in Japanese pear (Pyrus pyrifolia)

    PubMed Central

    Nakajima, Ikuko; Sato, Yoshihiko; Saito, Toshihiro; Moriguchi, Takaya; Yamamoto, Toshiya

    2013-01-01

    Genetic transformation was successfully established producing both transformed adventitious shoots and calli in Japanese pear (Pyrus pyrifolia Nakai) by using cotyledons as explants. Cotyledons of five cultivars were co-cultivated with Agrobacterium tumefaciens strain LBA4404 carrying the pBIN19-sgfp, which contained a green fluorescent protein gene and the neomycin phosphotransferase gene. In order to increase transformation efficiency, sonication and ethylenedioxybis (ethylamine)-N,N,N′,N′-tetraacetic acid (EGTA) treatments were applied, which could produce physical wounds across the tissue and prevent plant defense reaction, respectively. Green fluorescent protein (GFP) fluorescence was evaluated two weeks and five months after Agrobacterium inoculation as measures of transient and stable transformations, respectively. As a result, sonication significantly increased both transient and stable expression of GFP fluorescence, whereas EGTA treatment did not show a positive effect on either. Out of 18 regenerated plantlets obtained, one plant regenerated from ‘Agenosho Shinanashi’ showed stable GFP fluorescence. This plant was confirmed as a transformant by PCR and genomic Southern blotting. Three other transformed regenerated shoots by myb gene showed red color, which were derived from ‘Imamuraaki’ by the same transformation method. Transformation system in this study was shown to be reproducible since plural transformants were obtained. PMID:24273422

  15. Improved Agrobacterium-mediated transformation and high efficiency of root formation from hypocotyl meristem of spring Brassica napus 'Precocity' cultivar.

    PubMed

    Liu, X X; Lang, S R; Su, L Q; Liu, X; Wang, X F

    2015-01-01

    Rape seed (Brassica napus L.) is one of the most important oil seed crops in the world. Genetic manipulation of rapeseed requires a suitable tissue culture system and an efficient method for plant regeneration, as well as an efficient transformation procedure. However, development of transgenic B. napus has been problematic, and current studies are limited to cultivated varieties. In this study, we report a protocol for regeneration of transgenic rape after Agrobacterium-mediated transformation of hypocotyls from the spring B. napus 'Precocity' cultivar. We analyzed the effects of plant growth regulators in the medium on regeneration. Additionally, factors affecting the transformation efficiency, including seedling age, Agrobacterium concentration, infection time, and co-cultivation time, were assessed by monitoring GUS expression. Results from these experiments revealed that transformation was optimized when the meristematic parts of the hypocotyls were taken from 8 day-old seedlings, cultured on Murashinge and Skoog basal media containing 0.1 mg/L 1-naphthaleneacetic acid and 2.5 mg/L 6-benzylaminopurine, and incubated in Agrobacterium suspension (OD600 = 0.5) for 3 to 5 min, followed by 2 days of co-cultivation. Integration of T-DNA into the plant genome was confirmed by polymerase chain reaction (PCR), b-glucuronidase histochemical staining, and quantitative real-time PCR. The protocols developed for regeneration, transformation, and rooting described in this study could help to accelerate the development of transgenic spring rape varieties with novel features. PMID:26681030

  16. Development of an Efficient Agrobacterium-Mediated Transformation System and Production of Herbicide-Resistant Transgenic Plants in Garlic (Allium sativum L.)

    PubMed Central

    Ahn, Yul-Kyun; Yoon, Moo-Kyoung; Jeon, Jong-Seong

    2013-01-01

    The genetic improvement of garlic plants (Allium sativum L.) with agronomical beneficial traits is rarely achieved due to the lack of an applicable transformation system. Here, we developed an efficient Agrobacterium-mediated transformation procedure with Danyang, an elite Korean garlic cultivar. Examination of sGFP (synthetic green fluorescence protein) expression revealed that treatment with 2-(N-morpholino) ethanesulfonic acid (MES), L-cysteine and/or dithiothreitol (DTT) gives the highest efficiency in transient gene transfer during Agrobacterium co-cultivation with calli derived from the roots of in vitro plantlets. To increase stable transformation efficiency, a two-step selection was employed on the basis of hygromycin resistance and sGFP expression. Of the hygromycin-resistant calli initially produced, only sGFP-expressing calli were subcultured for selection of transgenic calli. Transgenic plantlets produced from these calli were grown to maturity. The transformation efficiency increased up to 10.6% via our optimized procedure. DNA and RNA gel-blot analysis indicated that transgenic garlic plants stably integrated and expressed the phosphinothricin acetyltransferase (PAT) gene. A herbicide spraying assay demonstrated that transgenic plants of garlic conferred herbicide resistance, whilst non-transgenic plants and weeds died. These results indicate that our transformation system can be efficiently utilized to produce transgenic garlic plants with agronomic benefits. PMID:23832764

  17. Development of an efficient Agrobacterium-mediated transformation system and production of herbicide-resistant transgenic plants in garlic (Allium sativum L.).

    PubMed

    Ahn, Yul-Kyun; Yoon, Moo-Kyoung; Jeon, Jong-Seong

    2013-08-01

    The genetic improvement of garlic plants (Allium sativum L.) with agronomical beneficial traits is rarely achieved due to the lack of an applicable transformation system. Here, we developed an efficient Agrobacterium-mediated transformation procedure with Danyang, an elite Korean garlic cultivar. Examination of sGFP (synthetic green fluorescence protein) expression revealed that treatment with 2-(N-morpholino) ethanesulfonic acid (MES), L-cysteine and/or dithiothreitol (DTT) gives the highest efficiency in transient gene transfer during Agrobacterium co-cultivation with calli derived from the roots of in vitro plantlets. To increase stable transformation efficiency, a two-step selection was employed on the basis of hygromycin resistance and sGFP expression. Of the hygromycin-resistant calli initially produced, only sGFP-expressing calli were subcultured for selection of transgenic calli. Transgenic plantlets produced from these calli were grown to maturity. The transformation efficiency increased up to 10.6% via our optimized procedure. DNA and RNA gel-blot analysis indicated that transgenic garlic plants stably integrated and expressed the phosphinothricin acetyltransferase (PAT) gene. A herbicide spraying assay demonstrated that transgenic plants of garlic conferred herbicide resistance, whilst nontransgenic plants and weeds died. These results indicate that our transformation system can be efficiently utilized to produce transgenic garlic plants with agronomic benefits. PMID:23832764

  18. Cadophora finlandia and Phialocephala fortinii: Agrobacterium-mediated transformation and functional GFP expression.

    PubMed

    Gorfer, Markus; Klaubauf, Sylvia; Bandian, Dragana; Strauss, Joseph

    2007-07-01

    Hygromycin B resistance was transferred to the sterile mycelia of Cadophora finlandia and Phialocephala fortinii by co-cultivation with Agrobacterium tumefaciens. Constitutively expressed green fluorescent protein (GFP) was also introduced using the same vector. Confocal laser scanning microscopy (CLSM) revealed strong fluorescence of transformants. Both traits were mitotically stable during one year of subculturing on non-selective growth medium. Southern blot analysis showed that the majority of the transformants contained single-copy integrations at random sites in the genome. PMID:17662587

  19. Female Reproductive Tissues Are the Primary Target of Agrobacterium-Mediated Transformation by the Arabidopsis Floral-Dip Method1

    PubMed Central

    Desfeux, Christine; Clough, Steven J.; Bent, Andrew F.

    2000-01-01

    The floral-dip method for Agrobacterium-mediated transformation of Arabidopsis allows efficient plant transformation without need for tissue culture. To facilitate use with other plant species, we investigated the mechanisms that underlie this method. In manual outcrossing experiments, application of Agrobacterium tumefaciens to pollen donor plants did not produce any transformed progeny, whereas application of Agrobacterium to pollen recipient plants yielded transformants at a rate of 0.48%. Agrobacterium strains with T-DNA carrying gusA (encoding β-glucuronidase [GUS]) under the control of 35S, LAT52, or ACT11 promoters revealed delivery of GUS activity to developing ovules, whereas no GUS staining of pollen or pollen tubes was observed. Transformants derived from the same seed pod contained independent T-DNA integration events. In Arabidopsis flowers, the gynoecium develops as an open, vase-like structure that fuses to form closed locules roughly 3 d prior to anthesis. In correlation with this fact, we found that the timing of Agrobacterium infection was critical. Transformants were obtained and GUS staining of ovules and embryo sacs was observed only if the Agrobacterium were applied 5 d or more prior to anthesis. A 6-fold higher rate of transformation was obtained with a CRABS-CLAW mutant that maintains an open gynoecium. Our results suggest that ovules are the site of productive transformation in the floral-dip method, and further suggest that Agrobacterium must be delivered to the interior of the developing gynoecium prior to locule closure if efficient transformation is to be achieved. PMID:10889238

  20. Development of Agrobacterium-mediated transformation of highly valued hill banana cultivar Virupakshi (AAB) for resistance to BBTV disease.

    PubMed

    Elayabalan, Sivalingam; Kalaiponmani, Kalaimughilan; Subramaniam, Sreeramanan; Selvarajan, Ramasamy; Panchanathan, Radha; Muthuvelayoutham, Ramlatha; Kumar, Krish K; Balasubramanian, Ponnuswami

    2013-04-01

    One of the most severe viral diseases of hill banana is caused by banana bunchy top virus (BBTV), a nanovirus transmitted by the aphid Pentalonia nigronervosa. In this study, we reported the Agrobacterium-mediated transformation on a highly valued hill banana cultivar Virupakshi (AAB) for resistance to BBTV disease. The target of the RNA interference (RNAi) is the rep gene, encoded by the BBTV-DNA1. In order to develop RNAi construct targeting the BBTV rep gene, the full-length rep gene of 870 bp was polymerase chain reaction amplified from BBTV infected hill banana sample DNA, cloned and confirmed by DNA sequencing. The partial rep gene fragment was cloned in sense and anti sense orientation in the RNAi intermediate vector, pSTARLING-A. After cloning in pSTARLING-A, the cloned RNAi gene cassette was released by NotI enzyme digestion and cloned into the NotI site of binary vector, pART27. Two different explants, embryogenic cells and embryogenic cell suspension derived microcalli were used for co-cultivation. Selection was done in presence of 100 mg/L kanamycin. In total, 143 putative transgenic hill banana lines were generated and established in green house condition. The presence of the transgenes was confirmed in the selected putative transgenic hill banana lines by PCR and reverse transcription PCR analyses. Transgenic hill banana plants expressing RNAi-BBTV rep were obtained and shown to resist infection by BBTV. The transformed plants are symptomless, and the replication of challenge BBTV almost completely suppressed. Hence, the RNAi mediating resistances were shown to be effective management of BBTV in hill banana. PMID:23184576

  1. Agrobacterium-mediated transformation of Eucalyptus globulus using explants with shoot apex with introduction of bacterial choline oxidase gene to enhance salt tolerance.

    PubMed

    Matsunaga, Etsuko; Nanto, Kazuya; Oishi, Masatoshi; Ebinuma, Hiroyasu; Morishita, Yoshihiko; Sakurai, Nozomu; Suzuki, Hideyuki; Shibata, Daisuke; Shimada, Teruhisa

    2012-01-01

    Eucalyptus globulus is one of the most economically important plantation hardwoods for paper making. However, its low transformation frequency has prevented genetic engineering of this species with useful genes. We found the hypocotyl section with a shoot apex has the highest regeneration ability among another hypocotyl sections, and have developed an efficient Agrobacterium-mediated transformation method using these materials. We then introduced a salt tolerance gene, namely a bacterial choline oxidase gene (codA) with a GUS reporter gene, into E. globulus. The highest frequency of transgenic shoot regeneration from hypocotyls with shoot apex was 7.4% and the average frequency in four experiments was 4.0%, 12-fold higher than that from hypocotyls without shoot apex. Using about 10,000 explants, over 250 regenerated buds were confirmed as transformants by GUS analysis. Southern blot analysis of 100 elongated shoots confirmed successful generation of stable transformants. Accumulation of glycinebetaine was investigated in 44 selected transgenic lines, which showed 1- to 12-fold higher glycinebetaine levels than non-transgenic controls. Rooting of 16 transgenic lines was successful using a photoautotrophic method under enrichment with 1,000 ppm CO(2). The transgenic whole plantlets were transplanted into potting soil and grown normally in a growth room. They showed salt tolerance to 300 mM NaCl. The points of our system are using explants with shoot apex as materials, inhibiting the elongation of the apex on the selection medium, and regenerating transgenic buds from the side opposite to the apex. This approach may also solve transformation problems in other important plants. PMID:22009051

  2. Genetic transformation of Metroxylon sagu (Rottb.) cultures via Agrobacterium-mediated and particle bombardment.

    PubMed

    Ibrahim, Evra Raunie; Hossain, Md Anowar; Roslan, Hairul Azman

    2014-01-01

    Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance. PMID:25295258

  3. Genetic Transformation of Metroxylon sagu (Rottb.) Cultures via Agrobacterium-Mediated and Particle Bombardment

    PubMed Central

    Ibrahim, Evra Raunie

    2014-01-01

    Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance. PMID:25295258

  4. Agrobacterium-mediated genetic transformation and plant regeneration of the hardwood tree species Fraxinus profunda.

    PubMed

    Stevens, Micah E; Pijut, Paula M

    2014-06-01

    This transformation and regeneration protocol provides an integral framework for the genetic improvement of Fraxinus profunda (pumpkin ash) for future development of plants resistant to the emerald ash borer. Using mature hypocotyls as the initial explants, an Agrobacterium tumefaciens-mediated genetic transformation system was successfully developed for pumpkin ash (Fraxinus profunda). This transformation protocol is an invaluable tool to combat the highly aggressive, non-native emerald ash borer (EAB), which has the potential to eliminate native Fraxinus spp. from the natural landscape. Hypocotyls were successfully transformed with Agrobacterium strain EHA105 harboring the pq35GR vector, containing an enhanced green fluorescent protein (EGFP) as well as a fusion gene between neomycin phosphotransferase (nptII) and gusA. Hypocotyls were cultured for 7 days on Murashige and Skoog (MS) medium with 22.2 μM 6-benzyladenine (BA), 4.5 μM thidiazuron (TDZ), 50 mg L(-1) adenine hemisulfate (AS), and 10 % coconut water (CW) prior to transformation. Hypocotyls were transformed using 90 s sonication plus 10 min vacuum infiltration after Agrobacterium was exposed to 100 μM acetosyringone for 1 h. Adventitious shoots were regenerated on MS medium with 22.2 μM BA, 4.5 μM TDZ, 50 mg L(-1) AS, 10 % CW, 400 mg L(-1) timentin, and 20 mg L(-1) kanamycin. Timentin at 400 and 20 mg L(-1) kanamycin were most effective at controlling Agrobacterium growth and selecting for transformed cells, respectively. The presence of nptII, GUS (β-glucuronidase), and EGFP in transformed plants was confirmed using polymerase chain reaction (PCR), while the expression of EGFP was also confirmed through fluorescent microscopy and reverse transcription-PCR. This transformation protocol provides an integral foundation for future genetic modifications of F. profunda to provide resistance to EAB. PMID:24493252

  5. Agrobacterium-mediated genetic transformation of Pogostemon cablin (Blanco) Benth. Using leaf explants: bactericidal effect of leaf extracts and counteracting strategies.

    PubMed

    Paul, Anamika; Bakshi, Souvika; Sahoo, Debee Prasad; Kalita, Mohan Chandra; Sahoo, Lingaraj

    2012-04-01

    An optimized protocol for Agrobacterium tumefaciens-mediated transformation of patchouli using leaf disk explants is reported. In vitro antibacterial activity of leaf extracts of the plants revealed Agrobacterium sensitivity to the extracts. Fluorometric assay of bacterial cell viability indicated dose-dependent cytotoxic activity of callus extract against Agrobacterium cells. Addition of 0.1% Tween 20 and 2 g/l L-glutamine to Agrobacterium infection medium counteracted the bactericidal effect and significantly increased the T-DNA delivery to explants. A short preculture of explants for 2 days followed by infection with Agrobacterium in medium containing 150 μM of acetosyringone were found essential for efficient T-DNA delivery. Cocultivation for 3 days at 22 °C in conjunction with other optimized factors resulted in maximum T-DNA delivery. The Agrobacterium-mediated transformation of leaf disk explants were found significantly related to physiological age of the explants, age and origin of the of the donor plant. Leaf explants from second node of the 3-month-old in vivo plants showed highest transformation efficiency (94.3%) revealed by transient GUS expression assay. Plants selected on medium containing 20 mg/l kanamycin showed stable GUS expression in leaves and stem. The elongated shoots readily developed roots on kanamycin-free rooting medium and on transfer to soil, plants were successfully established. Polymerase chain reaction (PCR) and reverse-transcriptase PCR analysis in putative plants confirmed their transgenic nature. The established transformation method should provide new opportunities for the genetic improvement of patchouli for desirable trait. PMID:22434351

  6. Review of methodologies and a protocol for the Agrobacterium-mediated transformation of wheat

    PubMed Central

    Jones, Huw D; Doherty, Angela; Wu, Huixia

    2005-01-01

    Since the first report of wheat transformation by Agrobacterium tumefaciens in 1997, various factors that influence T-DNA delivery and regeneration in tissue culture have been further investigated and modified. This paper reviews the current methodology literature describing Agrobacterium transformation of wheat and provides a complete protocol that we have developed and used to produce over one hundred transgenic lines in both spring and winter wheat varieties. PMID:16270934

  7. Review of methodologies and a protocol for the Agrobacterium-mediated transformation of wheat.

    PubMed

    Jones, Huw D; Doherty, Angela; Wu, Huixia

    2005-09-01

    Since the first report of wheat transformation by Agrobacterium tumefaciens in 1997, various factors that influence T-DNA delivery and regeneration in tissue culture have been further investigated and modified. This paper reviews the current methodology literature describing Agrobacterium transformation of wheat and provides a complete protocol that we have developed and used to produce over one hundred transgenic lines in both spring and winter wheat varieties. PMID:16270934

  8. Successful Agrobacterium mediated transformation of Thielaviopsis basicola by optimizing multiple conditions.

    PubMed

    Tzima, Aliki K; Paplomatas, Epaminondas J; Schoina, Charikleia; Domazakis, Emmanouil; Kang, Seogchan; Goodwin, Paul H

    2014-08-01

    Thielaviopsis basicola is a hemibiotrophic root pathogen causing black root rot in a wide range of economically important crops. Our initial attempts to transform T. basicola using standard Agrobacterium tumefaciens-mediated transformation (ATMT) protocols were unsuccessful. Successful transformation required the addition of V8 juice (to induce germination of T. basicola chlamydospores) and higher concentrations of acetosyringone in the co-cultivation medium, and of chlamydospores/endoconidia, A. tumefaciens cells during co-cultivation. With these modifications, two T. basicola strains were successfully transformed with the green (egfp) or red (AsRed) fluorescent protein genes. Chlamydospores/endoconidia transformed with the egfp gene exhibited strong green fluorescence, but their fluorescence became weaker as the germ tubes emerged. Transformants harbouring the AsRed gene displayed strong red fluorescence in both chlamydospores/endoconidia and germ tubes. Fluorescent microscopic observations of an AsRed-labelled strain colonizing roots of transgenic Nicotiana benthamiana plants, which express the actin filaments labelled with EGFP, at 24 hours post inoculation showed varying levels of fungal germination and penetration. At this stage, the infection appeared to be biotrophic with the EGFP-labelled host actin filaments not being visibly degraded, even in host root cells in close contact with the hyphae. This is the first report of ATMT of T. basicola, and the use of an AsRed-labelled strain to directly observe the root infection process. PMID:25110130

  9. Agrobacterium-mediated transformation of promising oil-bearing marine algae Parachlorella kessleri.

    PubMed

    Rathod, Jayant Pralhad; Prakash, Gunjan; Pandit, Reena; Lali, Arvind M

    2013-11-01

    Parachlorella kessleri is a unicellular alga which grows in fresh as well as marine water and is commercially important as biomass/lipid feedstock and in bioremediation. The present study describes the successful transformation of marine P. kessleri with the help of Agrobacterium tumefaciens. Transformed marine P. kessleri was able to tolerate more than 10 mg l(-1) hygromycin concentration. Co-cultivation conditions were modulated to allow the simultaneous growth of both marine P. kessleri and A. tumefaciens. For co-cultivation, P. kessleri was shifted from Walne's to tris acetate phosphate medium to reduce the antibiotic requirement during selection. In the present study, the transfer of T-DNA was successful without using acetosyringone. Biochemical and genetic analyses were performed for expression of transgenes by GUS assay and PCR in transformants. Establishment of this protocol would be useful in further genetic modification of oil-bearing Parachlorella species. PMID:24097049

  10. Yeast Actin-Related Protein ARP6 Negatively Regulates Agrobacterium-Mediated Transformation of Yeast Cell.

    PubMed

    Luo, Yumei; Chen, Zikai; Zhu, Detu; Tu, Haitao; Pan, Shen Quan

    2015-01-01

    The yeasts, including Saccharomyces cerevisiae and Pichia pastoris, are single-cell eukaryotic organisms that can serve as models for human genetic diseases and hosts for large scale production of recombinant proteins in current biopharmaceutical industry. Thus, efficient genetic engineering tools for yeasts are of great research and economic values. Agrobacterium tumefaciens-mediated transformation (AMT) can transfer T-DNA into yeast cells as a method for genetic engineering. However, how the T-DNA is transferred into the yeast cells is not well established yet. Here our genetic screening of yeast knockout mutants identified a yeast actin-related protein ARP6 as a negative regulator of AMT. ARP6 is a critical member of the SWR1 chromatin remodeling complex (SWR-C); knocking out some other components of the complex also increased the transformation efficiency, suggesting that ARP6 might regulate AMT via SWR-C. Moreover, knockout of ARP6 led to disruption of microtubule integrity, higher uptake and degradation of virulence proteins, and increased DNA stability inside the cells, all of which resulted in enhanced transformation efficiency. Our findings have identified molecular and cellular mechanisms regulating AMT and a potential target for enhancing the transformation efficiency in yeast cells. PMID:26425545

  11. Yeast Actin-Related Protein ARP6 Negatively Regulates Agrobacterium-Mediated Transformation of Yeast Cell

    PubMed Central

    Luo, Yumei; Chen, Zikai; Zhu, Detu; Tu, Haitao; Pan, Shen Quan

    2015-01-01

    The yeasts, including Saccharomyces cerevisiae and Pichia pastoris, are single-cell eukaryotic organisms that can serve as models for human genetic diseases and hosts for large scale production of recombinant proteins in current biopharmaceutical industry. Thus, efficient genetic engineering tools for yeasts are of great research and economic values. Agrobacterium tumefaciens-mediated transformation (AMT) can transfer T-DNA into yeast cells as a method for genetic engineering. However, how the T-DNA is transferred into the yeast cells is not well established yet. Here our genetic screening of yeast knockout mutants identified a yeast actin-related protein ARP6 as a negative regulator of AMT. ARP6 is a critical member of the SWR1 chromatin remodeling complex (SWR-C); knocking out some other components of the complex also increased the transformation efficiency, suggesting that ARP6 might regulate AMT via SWR-C. Moreover, knockout of ARP6 led to disruption of microtubule integrity, higher uptake and degradation of virulence proteins, and increased DNA stability inside the cells, all of which resulted in enhanced transformation efficiency. Our findings have identified molecular and cellular mechanisms regulating AMT and a potential target for enhancing the transformation efficiency in yeast cells. PMID:26425545

  12. Hypocotyl-based Agrobacterium-mediated transformation of soybean (Glycine max) and application for RNA interference.

    PubMed

    Wang, Geliang; Xu, Yinong

    2008-07-01

    An efficient system of gene transformation is necessary for soybean [Glycine max (L.) Merrill] functional genomics and gene modification by using RNA interference (RNAi) technology. To establish such system, we improved the conditions of tissue culture and transformation for increasing the frequency of adventitious shoots and decreasing the browning and necrosis of hypocotyls. Adding N(6)-benzylaminopurine (BAP) and silver nitrate in culture medium enhanced the shoot formation on hypocotyls. BAP increased the frequency of the hypocotyls containing adventitious shoots, while silver nitrate increased the number of shoots on the hypocotyls. As a result, the number of adventitious shoots on hypocotyls cultured in medium containing both BAP and silver nitrate was 5-fold higher than the controls. Adding antioxidants in co-cultivation medium resulted in a significant decrease in occurrence of browning and necrosis of hypocotyls and increase in levels of beta-Glucuronidase (GUS) gene expression. Histochemical assays showed that the apical meristem of hypocotyls was the "target tissue" for Agrobacterium tumefaciens transformation of soybean. Gene silencing of functional gene by using RNAi technology was carried out under above conditions. A silencing construct containing an inverted-repeat fragment of the GmFAD2 gene was introduced into soybean by using the A. tumefaciens-mediated transformation. Several lines with high oleic acid were obtained, in which mean oleic acid content ranged from 71.5 to 81.9%. Our study demonstrates that this transgenic approach could be efficiently used to improve soybean quality and productivity through functional genomics. PMID:18347801

  13. Enhanced Agrobacterium-mediated transformation of embryogenic calli of upland cotton.

    PubMed

    Zhang, Tianzhen; Wu, Shen-Jie

    2012-01-01

    Agrobacterium tumefaciens-mediated transformation of cotton embryogenic calli (EC) was enhanced by choosing appropriate EC and improving efficiency of coculture, selection cultivation, and plant regeneration. The binary vector pBI121 (containing a neomycin phosphotransferase II gene npt-II as a selection marker and a uidA gene as a reporter gene) was used to research transformation efficiency. After 48 h cocultivation, the number of β-glucuronidase (GUS)-positive calli characterized by yellow, loose, and fine-grained EC was twofold greater than that of gray, brown, and coarse granule EC. It indicated that the efficiency of transient transformation was affected by EC morphology. Transient transformation efficiency also was improved by cocultivation on the medium by adding 50 mg/L acetosyringone at 19°C for 48 h. Subculturing EC on the selection medium with low cell density increased the production of kanamycin-resistant (Km-R) calli lines. From an original 0.3 g EC, an average of 20 Km-R calli lines were obtained from a selection dish, and the GUS-positive rate of Km-R clones was 81.97%. A large number of normal plants were rapidly regenerated on the differentiation medium with dehydration treatments, and the GUS-positive rate of regeneration plants was about 72.6%. Polymerase chain reaction analysis of GUS-positive plantlets revealed a 100% positive detection rate for neomycin phosphotransferase II gene and gus gene. Southern blot of transgenic plants regenerated from different Km-R calli lines demonstrated that the target gene, mostly with the low copy number, was integrated into the cotton genome. PMID:22351014

  14. Expression and genomic integration of transgenes after Agrobacterium-mediated transformation of mature barley embryos.

    PubMed

    Uçarlı, C; Tufan, F; Gürel, F

    2015-01-01

    Mature embryos in tissue cultures are advantageous because of their abundance and rapid germination, which reduces genomic instability problems. In this study, 2-day-old isolated mature barley embryos were infected with 2 Agrobacterium hypervirulent strains (AGL1 and EHA105), followed by a 3-day period of co-cultivation in the presence of L-cystein amino acid. Chimeric expression of the b-glucuronidase gene (gusA) directed by a viral promoter of strawberry vein banding virus was observed in coleoptile epidermal cells and seminal roots in 5-day-old germinated seedlings. In addition to varying infectivity patterns in different strains, there was a higher ratio of transient b-glucuronidase expression in developing coleoptiles than in embryonic roots, indicating the high competency of shoot apical meristem cells in the mature embryo. A total of 548 explants were transformed and 156 plants developed to maturity on G418 media after 18-25 days. We detected transgenes in 74% of the screened plant leaves by polymerase chain reaction, and 49% of these expressed neomycin phosphotransferase II gene following AGL1 transformation. Ten randomly selected T0 transformants were analyzed using thermal asymmetric interlaced polymerase chain reaction and 24 fragments ranged between 200-600 base pairs were sequenced. Three of the sequences flanked with transferred-DNA showed high similarity to coding regions of the barley genome, including alpha tubulin5, homeobox 1, and mitochondrial 16S genes. We observed 70-200-base pair filler sequences only in the coding regions of barley in this study. PMID:25730049

  15. Agrobacterium-mediated genetic transformation of commercially elite rice restorer line using nptII gene as a plant selection marker.

    PubMed

    Chakraborty, M; Sairam Reddy, P; Laxmi Narasu, M; Krishna, Gaurav; Rana, Debashis

    2016-01-01

    Transformation of commercially important indica cultivars remains challenging for the scientific community even though Agrobacterium-mediated transformation protocols for a few indica rice lines have been well established. We report successful transformation of a commercially important restorer line JK1044R of indica rice hybrid JKRH 401. While following existing protocol, we optimized several parameters for callusing, regeneration and genetic transformation of JK1044R. Calli generated from the rice scutellum tissue were used for transformation by Agrobacterium harboring pCAMBIA2201. A novel two tire selection scheme comprising of Geneticin (G418) and Paramomycin were deployed for selection of transgenic calli as well as regenerated plantlets that expressed neomycin phosphotransferase-II gene encoded by the vector. One specific combination of G418 (30 mg l(-1)) and Paramomycin (70 mg l(-1)) was very effective for calli selection. Transformed and selected calli were detected by monitoring the expression of the reporter gene uidA (GUS). Regenerated plantlets were confirmed through PCR analysis of nptII and gus genes specific primers as well as dot blot using gus gene specific as probe. PMID:27186018

  16. Transient down-regulation of the RNA silencing machinery increases efficiency of Agrobacterium-mediated transformation of Arabidopsis.

    PubMed

    Bilichak, Andriy; Yao, Youli; Kovalchuk, Igor

    2014-06-01

    Agrobacterium tumefaciens is a plant pathogen that is widely used in plant transformation. As the process of transgenesis includes the delivery of single-stranded T-DNA molecule, we hypothesized that transformation rate may negatively correlate with the efficiency of the RNA-silencing machinery. Using mutants compromised in either the transcriptional or post-transcriptional gene-silencing pathways, two inhibitors of stable transformation were revealed-AGO2 and NRPD1a. Furthermore, an immunoprecipitation experiment has shown that NRPD1, a subunit of Pol IV, directly interacts with Agrobacterium T-DNA in planta. Using the Tobacco rattle virus (TRV)--based virus-induced gene silencing (VIGS) technique, we demonstrated that the transient down-regulation of the expression of either AGO2 or NRPD1a genes in reproductive organs of Arabidopsis, leads to an increase in transformation rate. We observed a 6.0- and 3.5-fold increase in transformation rate upon transient downregulation of either AGO2 or NRPD1a genes, respectively. This is the first report demonstrating the increase in the plant transformation rate via VIGS-mediated transient down-regulation of the components of epigenetic machinery in reproductive tissue. PMID:24472037

  17. Sonication, Vacuum Infiltration and Thiol Compounds Enhance the Agrobacterium-Mediated Transformation Frequency of Withania somnifera (L.) Dunal

    PubMed Central

    Sivanandhan, Ganeshan; Kapil Dev, Gnajothi; Theboral, Jeevaraj; Selvaraj, Natesan; Ganapathi, Andy; Manickavasagam, Markandan

    2015-01-01

    In the present study, we have established a stable transformation protocol via Agrobacterium tumafacines for the pharmaceutically important Withania somnifera. Six day-old nodal explants were used for 3 day co-cultivation with Agrobacterium tumefaciens strain LBA4404 harbouring the vector pCAMIBA2301. Among the different injury treatments, sonication, vacuum infiltration and their combination treatments tested, a vacuum infiltration for 10 min followed by sonication for 10 sec with A. tumefaciens led to a higher transient GUS expression (84% explants expressing GUS at regenerating sites). In order to improve gene integration, thiol compounds were added to co-cultivation medium. A combined treatment of L-Cys at 100 mg/l, STS at 125 mg/l, DTT at 75 mg/l resulted in a higher GUS expression (90%) in the nodal explants. After 3 days of co-cultivation, the explants were subjected to three selection cycles with increasing concentrations of kanamycin [100 to 115 mg/l]. The integration and expression of gusA gene in T0 and T1 transgenic plants were confirmed by polymerase chain reaction (PCR), and Southern blott analysis. These transformed plants (T0 and T1) were fertile and morphologically normal. From the present investigation, we have achieved a higher transformation efficiency of (10%). Withanolides (withanolide A, withanolide B, withanone and withaferin A) contents of transformed plants (T0 and T1) were marginally higher than control plants. PMID:25927703

  18. Improved cotyledonary node method using an alternative explant derived from mature seed for efficient Agrobacterium-mediated soybean transformation.

    PubMed

    Paz, Margie M; Martinez, Juan Carlos; Kalvig, Andrea B; Fonger, Tina M; Wang, Kan

    2006-03-01

    The utility of transformation for soybean improvement requires an efficient system for production of stable transgenic lines. We describe here an improved cotyledonary node method using an alternative explant for Agrobacterium tumefaciens-mediated soybean transformation. We use the term "half-seed" to refer to this alternative cotyledonary explant that is derived from mature seed of soybean following an overnight imbibition and to distinguish it from cotyledonary node derived from 5-7-day-old seedlings. Transformation efficiencies using half-seed explants ranged between 1.4 and 8.7% with an overall efficiency of 3.8% based on the number of transformed events that have been confirmed in the T1 generation by phenotypic assay using the herbicide Liberty (active ingredient glufosinate) and by Southern analysis. This efficiency is 1.5-fold higher than the cotyledonary node method used in our laboratory. Significantly, the half-seed system is simple and does not require deliberate wounding of explants, which is a critical and technically demanding step in the cotyledonary node method. PMID:16249869

  19. A simple shoot multiplication procedure using internode explants, and its application for particle bombardment and Agrobacterium-mediated transformation in watercress.

    PubMed

    Ogita, Shinjiro; Usui, Miki; Shibutani, Nanae; Kato, Yasuo

    2009-07-01

    A shoot multiplication system derived from internode explants was investigated with the aim of improving genetic characteristics of watercress (Nasturtium officinale R. Br.). Internodes of ca. 1 cm excised from in vitro stock shoot culture were placed on half-strength Murashige and Skoog (MS) medium supplemented with 3 muM 2,4-dichlorophenoxyacetic acid as a pre-treatment. Laser scanning microscopy indicated clearly that the first sign of meristematic cell division could be seen after 1-2 days of pre-culture, and meristematic tissues multiplied along the vascular cambium of the internode segment during 7 days of culture. Multiple shoots could be obtained from more than 90% of the pre-treated explants when they were subsequently transferred to MS medium supplemented with 1 muM thidiazuron for 3 weeks. These findings indicate that pre-treatment of the internodes for 7 days promoted their capacity for organogenesis. Using this pre-treatment, frequent generation of transgenic watercress plants was achieved by adapting particle bombardment and Agrobacterium-mediated transformation techniques with a construct expressing a synthetic green florescent protein gene. PMID:19308313

  20. Highly efficient Agrobacterium-mediated transformation of embryogenic cell suspensions of Musa acuminata cv. Mas (AA) via a liquid co-cultivation system.

    PubMed

    Huang, Xia; Huang, Xue-Lin; Xiao, Wang; Zhao, Jie-Tang; Dai, Xue-Mei; Chen, Yun-Feng; Li, Xiao-Ju

    2007-10-01

    A high efficient protocol of Agrobacterium-mediated transformation of Musa acuminata cv. Mas (AA), a major banana variety of the South East Asia region, was developed in this study. Male-flower-derived embryogenic cell suspensions (ECS) were co-cultivated in liquid medium with Agrobacterium strain EHA105 harboring a binary vector pCAMBIA2301 carrying nptII and gusA gene in the T-DNA. Depending upon conditions and duration of co-cultivation in liquid medium, 0-490 transgenic plants per 0.5 ml packed cell volume (PCV) of ECS were obtained. The optimum duration of inoculation was 2 h, and the highest transformation frequency was achieved when infected ECS were co-cultivated in liquid medium first for 12 h at 40 rpm and then for 156 h at 100 rpm on a rotary shaker. Co-cultivation for a shorter duration (72 h) or shaking constantly at 100 rpm at the same duration gave 1.6 and 1.8 folds lower transformation efficiency, respectively. No transgenic plants were obtained in parallel experiments carried on semi-solid media. Histochemical GUS assay and molecular analysis in several tissues of the transgenic plants demonstrated that foreign genes were stably integrated into the banana genome. Compared to semi-solid co-cultivation transformation in other banana species, it is remarkable that liquid co-cultivation was much more efficient for transformation of the Mas cultivar, and was at least 1 month faster for regenerating transgenic plants. PMID:17551731

  1. Identifying a Carotenoid Cleavage Dioxygenase 4a Gene and Its Efficient Agrobacterium-Mediated Genetic Transformation in Bixa orellana L.

    PubMed

    Sankari, Mohan; Hemachandran, Hridya; Anantharaman, Amirtha; Babu, Subramanian; Madrid, Renata Rivera; C, George Priya Doss; Fulzele, Devanand P; Siva, Ramamoorthy

    2016-07-01

    Carotenoids are metabolized to apocarotenoids through the pathway catalysed by carotenoid cleavage oxygenases (CCOs). The apocarotenoids are economically important as it is known to have therapeutic as well as industrial applications. For instance, bixin from Bixa orellana and crocin from Crocus sativus are commercially used as a food colourant and cosmetics since prehistoric time. In our present study, CCD4a gene has been identified and isolated from leaves of B. orellana for the first time and named as BoCCD4a; phylogenetic analysis was carried out using CLUSTAL W. From sequence analysis, BoCCD4a contains two exons and one intron, which was compared with the selected AtCCD4, RdCCD4, GmCCD4 and CmCCD4a gene. Further, the BoCCD4a gene was cloned into pCAMBIA 1301, transformed into Agrobacterium tumefaciens EHA105 strain and subsequently transferred into hypocotyledons and callus of B. orellana by agro-infection. Selection of stable transformation was screened on the basis of PCR detection by using GUS and hptII specific primer, which was followed by histochemical characterization. The percent transient GUS expression in hypocotyledons and callus was 84.4 and 80 %, respectively. The expression of BoCCD4a gene in B. orellana was confirmed through RT-PCR analysis. From our results, the sequence analysis of BoCCD4a gene of B. orellana was closely related to the CsCCD4 gene of C. sativus, which suggests this gene may have a role in various processes such as fragrance, insect attractant and pollination. PMID:26922728

  2. Factors influencing somatic embryogenesis, regeneration, and Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) cultivar TME14.

    PubMed

    Nyaboga, Evans N; Njiru, Joshua M; Tripathi, Leena

    2015-01-01

    Routine production of large numbers of transgenic plants is required to fully exploit advances in cassava biotechnology and support development of improved germplasm for deployment to farmers. This article describes an improved, high-efficiency transformation protocol for recalcitrant cassava cultivar TME14 preferred in Africa. Factors that favor production of friable embryogenic calli (FEC) were found to be use of DKW medium, crushing of organized embryogenic structures (OES) through 1-2 mm sized metal wire mesh, washing of crushed OES tissues and short exposure of tyrosine to somatic embryos; and transformation efficiency was enhanced by use of low Agrobacterium density during co-cultivation, co-centrifugation of FEC with Agrobacterium, germination of paramomycin resistant somatic embryos on medium containing BAP with gradual increase in concentration and variations of the frequency of subculture of cotyledonary-stage embryos on shoot elongation medium. By applying the optimized parameters, FEC were produced for cassava cultivar TME14 and transformed using Agrobacterium strain LBA4404 harboring the binary vector pCAMBIA2301. About 70-80 independent transgenic lines per ml settled cell volume (SCV) of FEC were regenerated on selective medium. Histochemical GUS assays confirmed the expression of gusA gene in transformed calli, somatic embryos and transgenic plants. The presence and integration of the gusA gene were confirmed by PCR and Southern blot analysis, respectively. RT-PCR analysis of transgenic plants confirmed the expression of gusA gene. This protocol demonstrates significantly enhanced transformation efficiency over existing cassava transformation protocols and could become a powerful tool for functional genomics and transferring new traits into cassava. PMID:26113851

  3. Factors influencing somatic embryogenesis, regeneration, and Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) cultivar TME14

    PubMed Central

    Nyaboga, Evans N.; Njiru, Joshua M.; Tripathi, Leena

    2015-01-01

    Routine production of large numbers of transgenic plants is required to fully exploit advances in cassava biotechnology and support development of improved germplasm for deployment to farmers. This article describes an improved, high-efficiency transformation protocol for recalcitrant cassava cultivar TME14 preferred in Africa. Factors that favor production of friable embryogenic calli (FEC) were found to be use of DKW medium, crushing of organized embryogenic structures (OES) through 1–2 mm sized metal wire mesh, washing of crushed OES tissues and short exposure of tyrosine to somatic embryos; and transformation efficiency was enhanced by use of low Agrobacterium density during co-cultivation, co-centrifugation of FEC with Agrobacterium, germination of paramomycin resistant somatic embryos on medium containing BAP with gradual increase in concentration and variations of the frequency of subculture of cotyledonary-stage embryos on shoot elongation medium. By applying the optimized parameters, FEC were produced for cassava cultivar TME14 and transformed using Agrobacterium strain LBA4404 harboring the binary vector pCAMBIA2301. About 70–80 independent transgenic lines per ml settled cell volume (SCV) of FEC were regenerated on selective medium. Histochemical GUS assays confirmed the expression of gusA gene in transformed calli, somatic embryos and transgenic plants. The presence and integration of the gusA gene were confirmed by PCR and Southern blot analysis, respectively. RT-PCR analysis of transgenic plants confirmed the expression of gusA gene. This protocol demonstrates significantly enhanced transformation efficiency over existing cassava transformation protocols and could become a powerful tool for functional genomics and transferring new traits into cassava. PMID:26113851

  4. Development of an Agrobacterium-mediated transformation system for the cold-adapted fungi Pseudogymnoascus destructans and P. pannorum.

    PubMed

    Zhang, Tao; Ren, Ping; Chaturvedi, Vishnu; Chaturvedi, Sudha

    2015-08-01

    The mechanisms of cold adaptation by fungi remain unknown. This topic is of high interest due to the emergence of white-nose syndrome (WNS), a skin infection of hibernating bats caused by Pseudogymnoascus destructans (Pd). Recent studies indicated that apart from Pd, there is an abundance of other Pseudogymnoascus species in the hibernacula soil. We developed an Agrobacterium tumefaciens-mediated transformation (ATMT) system for Pd and a related fungus Pseudogymnoascus pannorum (Pp) to advance experimental studies. URE1 gene encoding the enzyme urease was used as an easy to screen marker to facilitate molecular genetic analyses. A Uracil-Specific Excision Reagent (USER) Friendly pRF-HU2 vector containing Pd or Pp ure1::hygromycin (HYG) disruption cassette was introduced into A. tumefaciens AGL-1 cells by electroporation and the resulting strains were co-cultivated with conidia of Pd or Pp for various durations and temperatures to optimize the ATMT system. Overall, 680 Pd (0.006%) and 1800 Pp (0.018%) transformants were obtained from plating of 10(7) conidia; their recoveries were strongly correlated with the length of the incubation period (96h for Pd; 72h for Pp) and with temperature (15-18°C for Pd; 25°C for Pp). The homologous recombination in transformants was 3.1% for Pd and 16.7% for Pp. The availability of a standardized ATMT system would allow future molecular genetic analyses of Pd and related cold-adapted fungi. PMID:26051491

  5. Increased Agrobacterium-mediated transformation and rooting efficiencies in canola (Brassica napus L.) from hypocotyl segment explants

    NASA Technical Reports Server (NTRS)

    Cardoza, V.; Stewart, C. N.

    2003-01-01

    An efficient protocol for the production of transgenic Brassica napus cv. Westar plants was developed by optimizing two important parameters: preconditioning time and co-cultivation time. Agrobacterium tumefaciens-mediated transformation was performed using hypocotyls as explant tissue. Two variants of a green fluorescent protein (GFP)-encoding gene--mGFP5-ER and eGFP--both under the constitutive expression of the cauliflower mosaic virus 35S promoter, were used for the experiments. Optimizing the preconditioning time to 72 h and co-cultivation time with Agrobacterium to 48 h provided the increase in the transformation efficiency from a baseline of 4% to 25%. With mGFP5-ER, the transformation rate was 17% and with eGFP it was 25%. Transgenic shoots were selected on 200 mg/l kanamycin. Rooting efficiency was 100% on half-strength Murashige and Skoog medium with 10 g/l sucrose and 0.5 mg/l indole butyric acid in the presence of kanamycin.

  6. Bacterial Transposons Are Co-Transferred with T-DNA to Rice Chromosomes during Agrobacterium-Mediated Transformation

    PubMed Central

    Kim, Sung-Ryul; An, Gynheung

    2012-01-01

    Agrobacterium tumefaciens is widely utilized for delivering a foreign gene into a plant’s genome. We found the bacterial transposon Tn5393 in transgenic rice plants. Analysis of the flanking sequences of the transferred-DNA (T-DNA) identified that a portion of the Tn5393 sequence was present immediately next to the end of the T-DNA. Because this transposon was present in A. tumefaciens strain LBA4404, but not in EHA105 and GV3101, our findings indicated that Tn5393 was transferred from LBA4404 into the rice genome during the transformation process. We also noted that another bacterial transposon, Tn5563, is present in transgenic plants. Analyses of 331 transgenic lines revealed that 26.0% carried Tn5393 and 2.1% contained Tn5563. In most of the lines, an intact transposon was integrated into the T-DNA and transferred to the rice chromosome. More than one copy of T-DNA was introduced into the plants, often at a single locus. This resulted in T-DNA repeats of normal and transposon-carrying T-DNA that generated deletions of a portion of the T-DNA, joining the T-DNA end to the bacterial transposon. Based on these data, we suggest that one should carefully select the appropriate Agrobacterium strain to avoid undesirable transformation of such sequences. PMID:22570148

  7. Agrobacterium-mediated transformation of the β-subunit gene in 7S globulin protein in soybean using RNAi technology.

    PubMed

    Qu, J; Liu, S Y; Wang, P W; Guan, S Y; Fan, Y G; Yao, D; Zhang, L; Dai, J L

    2016-01-01

    The objective of this study was to use RNA interference (RNAi) to improve protein quality and decrease anti-nutritional effects in soybean. Agrobacterium tumefaciens-mediated transformation was conducted using RNAi and an expression vector containing the 7S globulin β-subunit gene. The BAR gene was used as the selective marker and cotyledonary nodes of soybean genotype Jinong 27 were chosen as explant material. Regenerated plants were detected by molecular biology techniques. Transformation of the β-subunit gene in the 7S protein was detected by PCR, Southern blot, and q-PCR. Positive plants (10 T0, and 6 T1, and 13 T2) were tested by PCR. Hybridization bands were detected by Southern blot analysis in two of the T1 transgenic plants. RNAi expression vectors containing the soybean 7S protein β-subunit gene were successfully integrated into the genome of transgenic plants. qRT-PCR analysis in soybean seeds showed a clear decrease in expression of the soybean β-subunit gene. The level of 7S protein β-subunit expression in transgenic plants decreased by 77.5% as compared to that of the wild-type plants. This study has established a basis for the application of RNAi to improve the anti-nutritional effects of soybean. PMID:27173254

  8. In vitro regeneration and optimization of factors affecting Agrobacterium mediated transformation in Artemisia Pallens, an important medicinal plant.

    PubMed

    Alok, Anshu; Shukla, Vishnu; Pala, Zarna; Kumar, Jitesh; Kudale, Subhash; Desai, Neetin

    2016-04-01

    Artemisia pallens is an important medicinal plant. In-vitro regeneration and multiplication of A. pallens have been established using attached cotyledons. Different growth regulators were considered for regeneration of multiple shoots. An average of 36 shoots per explants were obtained by culturing attached cotyledons on Murashige and Skoog's medium containing 2 mg/L BAP and 0.1 mg/L NAA, after 45 days. The shoots were rooted best on half Murashige and Skoog's medium with respect to media containing 1 mg/L IBA or 1 mg/L NAA. Different parameters such as type of bacterial strains, OD600 of bacterial culture, co-cultivation duration, concentration of acetosyringone and explants type were optimized for transient expression of the reporter gene. Agrobacterium tumefaciens harbouring pCambia1301 plasmid carrying β-glucuronidase as a reporter gene and hygromycin phosphotransferase as plant selectable marker genes were used for genetic transformation of A. pallens. Hygromycin lethality test showed concentration of 15 mg/L were sufficient to inhibit the growth of attached cotyledons and multiple shoot buds of nontransgenics in selection media. Up to 83 % transient transformation was found when attached cotyledons were co-cultivated with Agrobacterium strain AGL1 for 2 days at 22 °C on shoot induction medium. The bacterial growth was eliminated by addition of cefotaxime (200 mg/L) in selection media. T0 transgenic plants were confirmed by GUS histochemical assay and further by polymerase chain reaction (PCR) using uidA and hpt gene specific primers. The study is useful in establishing technological improvement in A. pallens by genetic engineering. PMID:27436917

  9. Agrobacterium-mediated transformation of potato using PLRV-rep and PVY CP genes and assessment of replicase mediated resistance against natural infection of PLRV

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Replicase-and coat protein gene-mediated resistances against potato leafroll virus (PLRV) and potato virus Y (PVY), respectively, demonstrated to be an effective way of protecting potato against two major virus problems (PLRV & PVY) world-wide. Potato cultivar Desiree was transformed using Agrobacte...

  10. Agrobacterium-mediated transformation of tomato with rolB gene results in enhancement of fruit quality and foliar resistance against fungal pathogens.

    PubMed

    Arshad, Waheed; Haq, Ihsan-ul-; Waheed, Mohammad Tahir; Mysore, Kirankumar S; Mirza, Bushra

    2014-01-01

    Tomato (Solanum lycopersicum L.) is the second most important cultivated crop next to potato, worldwide. Tomato serves as an important source of antioxidants in human diet. Alternaria solani and Fusarium oxysporum cause early blight and vascular wilt of tomato, respectively, resulting in severe crop losses. The foremost objective of the present study was to generate transgenic tomato plants with rolB gene and evaluate its effect on plant morphology, nutritional contents, yield and resistance against fungal infection. Tomato cv. Rio Grande was transformed via Agrobacterium tumefaciens harbouring rolB gene of Agrobacterium rhizogenes. rolB. Biochemical analyses showed considerable improvement in nutritional quality of transgenic tomato fruits as indicated by 62% increase in lycopene content, 225% in ascorbic acid content, 58% in total phenolics and 26% in free radical scavenging activity. Furthermore, rolB gene significantly improved the defence response of leaves of transgenic plants against two pathogenic fungal strains A. solani and F. oxysporum. Contrarily, transformed plants exhibited altered morphology and reduced fruit yield. In conclusion, rolB gene from A. rhizogenes can be used to generate transgenic tomato with increased nutritional contents of fruits as well as improved foliar tolerance against fungal pathogens. PMID:24817272

  11. Agrobacterium-Mediated Transformation of Tomato with rolB Gene Results in Enhancement of Fruit Quality and Foliar Resistance against Fungal Pathogens

    PubMed Central

    Arshad, Waheed; Haq, Ihsan-ul-; Waheed, Mohammad Tahir; Mysore, Kirankumar S.; Mirza, Bushra

    2014-01-01

    Tomato (Solanum lycopersicum L.) is the second most important cultivated crop next to potato, worldwide. Tomato serves as an important source of antioxidants in human diet. Alternaria solani and Fusarium oxysporum cause early blight and vascular wilt of tomato, respectively, resulting in severe crop losses. The foremost objective of the present study was to generate transgenic tomato plants with rolB gene and evaluate its effect on plant morphology, nutritional contents, yield and resistance against fungal infection. Tomato cv. Rio Grande was transformed via Agrobacterium tumefaciens harbouring rolB gene of Agrobacterium rhizogenes. rolB. Biochemical analyses showed considerable improvement in nutritional quality of transgenic tomato fruits as indicated by 62% increase in lycopene content, 225% in ascorbic acid content, 58% in total phenolics and 26% in free radical scavenging activity. Furthermore, rolB gene significantly improved the defence response of leaves of transgenic plants against two pathogenic fungal strains A. solani and F. oxysporum. Contrarily, transformed plants exhibited altered morphology and reduced fruit yield. In conclusion, rolB gene from A. rhizogenes can be used to generate transgenic tomato with increased nutritional contents of fruits as well as improved foliar tolerance against fungal pathogens. PMID:24817272

  12. Comparison between Agrobacterium-mediated and direct gene transfer using the gene gun.

    PubMed

    Gao, Caixia; Nielsen, Klaus K

    2013-01-01

    Agrobacterium-mediated transformation and direct gene transfer using the gene gun (microparticle -bombardment) are the two most widely used methods for plant genetic modification. The Agrobacterium method has been successfully practiced in dicots for many years, but only recently have efficient protocols been developed for grasses. Microparticle bombardment has evolved as a method delivering exogenous nucleic acids into plant genome and is a commonly employed technique in plant science. Here these two systems are compared for transformation efficiency, transgene integration, and transgene expression when used to transform tall fescue (Festuca arundinacea Schreb.). The tall fescue transformation protocols lead to the production of large numbers of fertile, independent transgenic lines. PMID:23104329

  13. An efficient Agrobacterium-mediated transformation and regeneration system for leaf explants of two elite aspen hybrid clones Populus alba × P. berolinensis and Populus davidiana × P. bolleana.

    PubMed

    Wang, Haihai; Wang, Cuiting; Liu, Hua; Tang, Renjie; Zhang, Hongxia

    2011-11-01

    Transgenic technology has been successfully used for gene function analyses and trait improvement in cereal plants. However, its usage is limited in woody plants, especially in the difficult-to-transform but commercially viable hybrid poplar. In this work, an efficient regeneration and transformation system was established for the production of two hybrid aspen clones: Populus alba × P. berolinensis and Populus davidiana × P. bolleana. A plant transformation vector designed to express the reporter gene uidA, encoding β-glucuronidase (GUS), driven by the cauliflower mosaic virus 35S promoter, was used to detect transformation event at early stages of plant regeneration, and to optimize the parameters that may affect poplar transformation efficiency. Bacterium strain and age of leaf explant are two major factors that affect transformation efficiency. Addition of thidiazuron (TDZ) improved both regeneration and transformation efficiency. The transformation efficiency is approximately 9.3% for P. alba × P. berolinensis and 16.4% for P. davidiana × P. bolleana. Using this system, transgenic plants were usually produced in less than 1 month after co-cultivation. The growth characteristics and morphology of transgenic plants were identical to the untransformed wild type plants, and the transgenes could be inherited by vegetative propagation, as confirmed by PCR, Southern blotting, RT-PCR and β-glucuronidase staining analyses. The establishment of this system will help to facilitate the studies of gene functions in tree growth and development at a genome level, and as well as the introduction of some valuable traits in aspen breeding. PMID:21717184

  14. Competence of Immature Maize Embryos for Agrobacterium-Mediated Gene Transfer.

    PubMed Central

    Schlappi, M; Hohn, B

    1992-01-01

    Agrobacterium-mediated transfer of viral sequences to plant cells (agroinfection) was applied to study the susceptibility of immature maize embryos to the pathogen. The shoot apical meristem of immature embryos 10 to 20 days after pollination from four different maize genotypes was investigated for competence for agroinfection. There was a direct correlation between different morphological stages of the unwounded immature embryos and their competence for agroinfection. Agroinfection frequency was highest in the embryogenic line A188. All developmental stages tested showed Agrobacterium virulence gene-inducing activity, whereas bacteriocidal substances were produced at stages of the immature embryos competent for agroinfection. The results suggested that Agrobacterium may require differentiated tissue in the maize shoot apical meristem before wounding for successful T-DNA transfer. This requirement for the young maize embryo has implications for the possible use of Agrobacterium for maize transformation. PMID:12297627

  15. Agrobacterium-mediated transient MaFT expression in mulberry (Morus alba L.) leaves.

    PubMed

    Wu, Su-Li; Yang, Xiao-Bing; Liu, Li-Qun; Jiang, Tao; Wu, Hai; Su, Chao; Qian, Yong-Hua; Jiao, Feng

    2015-01-01

    To optimize Agrobacterium-mediated transient transformation assay in mulberry (Morus alba L.), various infiltration methods, Agrobacterium tumefaciens (A. tumefaciens) strains, and bacterial concentrations were tested in mulberry seedlings. Compared with LBA4404, GV3101 harboring pBE2133 plasmids presented stronger GUS signals at 3 days post infiltration using syringe. Recombinant plasmids pBE2133:GFP and pBE2133:GFP:MaFT were successfully constructed. Transient expression of MaFT:GFP protein was found in leaves, petiole (cross section), and shoot apical meristem (SAM) of mulberry according to the GFP signal. Moreover, MaFT:GFP mRNA was also detected in leaves and SAM via RT-PCR and qRT-PCR. An efficient transient transformation system could be achieved in mulberry seedlings by syringe using A. tumefaciens GV3101 at the OD600 of 0.5. The movement of MaFT expression from leaves to SAM might trigger the precocious flowering of mulberry. PMID:26024368

  16. Micro-shock Wave Assisted Plant Transformation

    NASA Astrophysics Data System (ADS)

    Gnanadhas, Divya Prakash; Datey, Akshay; Chakravortty, Dipshikha; Gopalan, Jagadeesh

    Genetically modified (GM) crops are developed by transforming the desired DNA to plant. There are various methods employed to achieve the required transformation in plants. Agrobacterium mediated transformation and Biolistics or particle bombardment method are the most commonly used methods.

  17. Agrobacterium-mediated gene transfer and enhanced green fluorescent protein visualization in the mycorrhizal ascomycete Tuber borchii: a first step towards truffle genetics.

    PubMed

    Grimaldi, Benedetto; de Raaf, Michiel A; Filetici, Patrizia; Ottonello, Simone; Ballario, Paola

    2005-07-01

    Mycorrhizal ascomycetes are ecologically and commercially important fungi that have proved impervious to genetic transformation so far. We report here on the successful transient transformation of Tuber borchii, an ectomycorrhizal ascomycete that colonizes a variety of trees and produces highly prized hypogeous fruitbodies known as "truffles". A hypervirulent Agrobacterium tumefaciens strain bearing the binary plasmid pBGgHg was used for transformation. The genes for hygromycin resistance and the enhanced green fluorescent protein (EGFP), both under the control of vector-borne promoters, were employed as selection markers. Patches of dark and fluorescent hyphae were observed upon fluorescence microscopic examination of hygromycin-resistant mycelia. The presence of EGFP was confirmed by both confocal microscopy and PCR analysis. The lack in the transformed mycelia of the DNA coding for kanamicin resistance (a trait encoded by a vector-borne gene located outside of the T-DNA region) indicates that Agrobacterium-mediated gene transfer correctly occurred in T. borchii. PMID:15868150

  18. Exploration of new perspectives and limitations in Agrobacterium mediated gene transfer technology. Progress report, [June 1, 1992-- May 31, 1994

    SciTech Connect

    Marton, L.

    1994-12-31

    This report describes progress aimed at constructing gene-transfer technology for Nicotiana plumbaginifolia. Most actual effort as described herein has so far been directed at exploring new perspectives and limitations in Agrobacterium mediated gene transfer. Accomplishments are described using a core homologous gene targeting vector.

  19. Development of Efficient Plant Regeneration and Transformation System for Impatiens Using Agrobacterium tumefaciens and Multiple Bud Cultures as Explants

    PubMed Central

    2010-01-01

    Background Impatiens (Impatiens walleriana) is a top selling floriculture crop. The potential for genetic transformation of Impatiens to introduce novel flower colors or virus resistance has been limited by its general recalcitrance to tissue culture and transformation manipulations. We have established a regeneration and transformation system for Impatiens that provides new alternatives to genetic improvement of this crop. Results In a first step towards the development of transgenic INSV-resistant Impatiens, we developed an efficient plant regeneration system using hypocotyl segments containing cotyledonary nodes as explants. With this regeneration system, 80% of explants produced an average of 32.3 elongated shoots per initial explant plated, with up to 167 elongated shoots produced per explant. Rooting efficiency was high, and 100% of shoots produced roots within 12 days under optimal conditions, allowing plant regeneration within approximately 8 weeks. Using this regeneration system, we developed an efficient Agrobacterium-mediated Impatiens transformation method using in vitro multiple bud cultures as explants and a binary plasmid (pHB2892) bearing gfp and nptII genes. Transgenic Impatiens plants, with a frequency up to 58.9%, were obtained within 12 to 16 weeks from inoculation to transfer of transgenic plants to soil. Transgenic plants were confirmed by Southern blot, phenotypic assays and T1 segregation analysis. Transgene expression was observed in leaves, stems, roots, flowers, and fruit. The transgenic plants were fertile and phenotypically normal. Conclusion We report the development of a simple and efficient Agrobacterium-mediated transformation system for Impatiens. To the best of our knowledge, there have been no reports of Agrobacterium-mediated transformation of Impatiens with experimental evidence of stable integration of T-DNA and of Agrobacterium-mediated transformation method for plants using in vitro maintained multiple bud cultures as

  20. Establishment of an Agrobacterium-mediated Inoculation System for Cucumber Green Mottle Mosaic Virus

    PubMed Central

    Kang, Minji; Seo, Jang-Kyun; Song, Dami; Choi, Hong-Soo; Kim, Kook-Hyung

    2015-01-01

    The infectious full-length cDNA clones of Cucumber green mottle mosaic virus (CGMMV) isolates KW and KOM, which were isolated from watermelon and oriental melon, respectively, were constructed under the control of the cauliflower mosaic virus 35S promoter. We successfully inoculated Nicotiana benthamiana with the cloned CGMMV isolates KW and KOM by Agrobacterium-mediated infiltration. Virulence and symptomatic characteristics of the cloned CGMMV isolates KW and KOM were tested on several indicator plants. No obvious differences between two cloned isolates in disease development were observed on the tested indicator plants. We also determined full genome sequences of the cloned CGMMV isolates KW and KOM. Sequence comparison revealed that only four amino acids (at positions 228, 699, 1212, and 1238 of the replicase protein region) differ between the cloned isolates KW and KOM. A previous study reported that the isolate KOM could not infect Chenopodium amaranticolor, but the cloned KOM induced chlorotic spots on the inoculated leaves. When compared with the previously reported sequence of the original KOM isolate, the cloned KOM contained one amino acid mutation (Ala to Thr) at position 228 of the replicase protein, suggesting that this mutation might be responsible for induction of chlorotic spots on the inoculated leaves of C. amaranticolor. PMID:26674677

  1. OPTIMIZING EFFICIENCY AND FLEXIBILITY OF AGROBACTERIUM-MEDIATED TRANSFORMATION OF RED CLOVER (TRIFOLIUM PRATENSE)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genetic modification of plants by the insertion of transgenes can be a powerful experimental approach to answer basic questions about gene product function. This technology can also be used to make improved crop varieties for use in the field. To apply this powerful tool to red clover (Trifolium pra...

  2. High-efficiency Agrobacterium-mediated transformation of Brachypodium distachyon inbred line Bd 21-3

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Brachypodium distachyon (Brachypodium) is a small grass with biological attributes (rapid generation time, small genome, diploid accessions, small stature and simple growth requirements) that make it suitable for use as a model system. In addition, a growing list of genomic resources have been devel...

  3. Spike-dip transformation of Setaria viridis.

    PubMed

    Saha, Prasenjit; Blumwald, Eduardo

    2016-04-01

    Traditional method of Agrobacterium-mediated transformation through the generation of tissue culture had limited success for Setaria viridis, an emerging C4 monocot model. Here we present an efficient in planta method for Agrobacterium-mediated genetic transformation of S. viridis using spike dip. Pre-anthesis developing spikes were dipped into a solution of Agrobacterium tumefaciens strain AGL1 harboring the β-glucuronidase (GUS) reporter gene driven by the cauliflower mosaic virus 35S (CaMV35S) promoter to standardize and optimize conditions for transient as well as stable transformations. A transformation efficiency of 0.8 ± 0.1% was obtained after dipping of 5-day-old S3 spikes for 20 min in Agrobacterium cultures containing S. viridis spike-dip medium supplemented with 0.025% Silwet L-77 and 200 μm acetosyringone. Reproducibility of this method was demonstrated by generating stable transgenic lines expressing β-glucuronidase plus (GUSplus), green fluorescent protein (GFP) and Discosoma sp. red fluorescent protein (DsRed) reporter genes driven by either CaMV35S or intron-interrupted maize ubiquitin (Ubi) promoters from three S. viridis genotypes. Expression of these reporter genes in transient assays as well as in T1 stable transformed plants was monitored using histochemical, fluorometric GUS activity and fluorescence microscopy. Molecular analysis of transgenic lines revealed stable integration of transgenes into the genome, and inherited transgenes expressed in the subsequent generations. This approach provides opportunities for the high-throughput transformation and potentially facilitates translational research in a monocot model plant. PMID:26932666

  4. Morphogenetic and chemical stability of long-term maintained Agrobacterium-mediated transgenic Catharanthus roseus plants.

    PubMed

    Verma, Priyanka; Sharma, Abhishek; Khan, Shamshad Ahmad; Mathur, Ajay Kumar; Shanker, Karuna

    2015-01-01

    Transgenic Catharanthus roseus plants (transgenic Dhawal [DT] and transgenic Nirmal [NT]) obtained from the Agrobacterium tumefaciens and Agrobacterium rhizognenes-mediated transformations, respectively, have been maintained in vitro for 5 years. Plants were studied at regular intervals for various parameters such as plant height, leaf size, multiplication rate, alkaloid profile and presence of marker genes. DT plant gradually lost the GUS gene expression and it was not detected in the fifth year while NT plant demonstrated the presence of genes rolA, rolB and rolC even in the fifth year, indicating the more stable nature of Ri transgene. Vindoline content in the DT was two times more than in non-transformed control plants. Alkaloid and tryptophan profiles were almost constant during the 5 years. The cluster analysis revealed that the DT plant is more close to the control Nirmal plant followed by NT plant. PMID:25102992

  5. Environment Control to Improve Recombinant Protein Yields in Plants Based on Agrobacterium-Mediated Transient Gene Expression.

    PubMed

    Fujiuchi, Naomichi; Matoba, Nobuyuki; Matsuda, Ryo

    2016-01-01

    Agrobacterium-mediated transient expression systems enable plants to produce a wide range of recombinant proteins on a rapid timescale. To achieve economically feasible upstream production and downstream processing, two yield parameters should be considered: (1) recombinant protein content per unit biomass and (2) recombinant protein productivity per unit area-time at the end of the upstream production. Because environmental factors in the upstream production have impacts on these parameters, environment control is important to maximize the recombinant protein yield. In this review, we summarize the effects of pre- and postinoculation environmental factors in the upstream production on the yield parameters and discuss the basic concept of environment control for plant-based transient expression systems. Preinoculation environmental factors associated with planting density, light quality, and nutrient supply affect plant characteristics, such as biomass and morphology, which in turn affect recombinant protein content and productivity. Accordingly, environment control for such plant characteristics has significant implications to achieve a high yield. On the other hand, postinoculation environmental factors, such as temperature, light intensity, and humidity, have been shown to affect recombinant protein content. Considering that recombinant protein production in Agrobacterium-mediated transient expression systems is a result of a series of complex biological events starting from T-DNA transfer from Agrobacterium tumefaciens to protein biosynthesis and accumulation in leaf tissue, we propose that dynamic environment control during the postinoculation process, i.e., changing environmental conditions at an appropriate timing for each event, may be a promising approach to obtain a high yield. Detailed descriptions of plant growth conditions and careful examination of environmental effects will significantly contribute to our knowledge to stably obtain high recombinant

  6. Environment Control to Improve Recombinant Protein Yields in Plants Based on Agrobacterium-Mediated Transient Gene Expression

    PubMed Central

    Fujiuchi, Naomichi; Matoba, Nobuyuki; Matsuda, Ryo

    2016-01-01

    Agrobacterium-mediated transient expression systems enable plants to produce a wide range of recombinant proteins on a rapid timescale. To achieve economically feasible upstream production and downstream processing, two yield parameters should be considered: (1) recombinant protein content per unit biomass and (2) recombinant protein productivity per unit area–time at the end of the upstream production. Because environmental factors in the upstream production have impacts on these parameters, environment control is important to maximize the recombinant protein yield. In this review, we summarize the effects of pre- and postinoculation environmental factors in the upstream production on the yield parameters and discuss the basic concept of environment control for plant-based transient expression systems. Preinoculation environmental factors associated with planting density, light quality, and nutrient supply affect plant characteristics, such as biomass and morphology, which in turn affect recombinant protein content and productivity. Accordingly, environment control for such plant characteristics has significant implications to achieve a high yield. On the other hand, postinoculation environmental factors, such as temperature, light intensity, and humidity, have been shown to affect recombinant protein content. Considering that recombinant protein production in Agrobacterium-mediated transient expression systems is a result of a series of complex biological events starting from T-DNA transfer from Agrobacterium tumefaciens to protein biosynthesis and accumulation in leaf tissue, we propose that dynamic environment control during the postinoculation process, i.e., changing environmental conditions at an appropriate timing for each event, may be a promising approach to obtain a high yield. Detailed descriptions of plant growth conditions and careful examination of environmental effects will significantly contribute to our knowledge to stably obtain high recombinant

  7. Stable Nuclear Transformation System for the Coccolithophorid Alga Pleurochrysis carterae

    PubMed Central

    Endo, Hirotoshi; Yoshida, Megumi; Uji, Toshiki; Saga, Naotsune; Inoue, Koji; Nagasawa, Hiromichi

    2016-01-01

    Of the three dominant marine microalgal groups, dinoflagellates and diatoms can undergo genetic transformation; however, no transformation method has been established for haptophytes to date. Here, we report the first stable genetic transformation of a coccolithophore, Pleurochrysis carterae, by means of polyethylene glycol (PEG)-mediated transfer of a bacterial hygromycin B-resistance gene. Together with the novel transient green fluorescent protein (GFP) expression system, this approach should facilitate further molecular-based research in this phylum. PMID:26947136

  8. In vitro plant regeneration and genetic transformation of Dichanthium annulatum.

    PubMed

    Kumar, Jitendra; Shukla, Sharad M; Bhat, Vishnu; Gupta, Sanjay; Gupta, M G

    2005-11-01

    Optimization of in vitro plant regeneration and genetic transformation of apomictic species such as Dichanthium annulatum would enable transfer of desirable genes. Seven genotypes of this grass species were screened through mature seed explant for embryogenic callus induction, callus growth and quality (color and texture), and shoot induction. Genotype IG-1999, which produced highly embryogenic, rapidly growing good-quality callus capable of regenerating at a high frequency, was selected for transformation experiments. Using a binary vector (pCAMBIA1305), frequency of GUS expression was compared between two methods of transformation. Bombardment of embryogenic calli with gold particles coated with pCAMBIA1305 at a distance of 11 cm, pressure of 4 bars, and vacuum of 27 Hg passing through 100 muM mesh produced maximum GUS expression (23%). Agrobacterium infection was maximum at an optical density of 2.0 when cocultured under vacuum for 15 min and cocultivated for 3 days at 28 degrees C in constant dark on MS medium of pH 5.8 with 3 mg/l 2,4-D, and 400 muM acetosyringone. Among two binary vectors used for Agrobacterium-mediated transformation, pCAMBIA1301 showed higher frequency of GUS expression while pCAMBIA1305 recorded more of the GUS spots per callus. Supplementation of acetosyringone in the cocultivation medium was found indispensable for Agrobacterium-mediated transformation. Injuring the calli through gold particle bombardment before their cocultivation with Agrobacterium improved the transformation efficiency. Several transgenic plants were developed using the PIG method, while stable GUS-expressing calli were multiplied during selection on MS medium containing 250 mg/l cefotaxime and 50 mg/l hygromycin, incubated in constant dark. A highly significant difference was observed between two methods of transformation for both frequency of GUS expression and GUS spots per callus. PIG-mediated transformation resulted in higher GUS expression compared to the

  9. Wheat (Triticum aestivum L.) transformation using mature embryos.

    PubMed

    Medvecká, Eva; Harwood, Wendy A

    2015-01-01

    In most protocols for the Agrobacterium-mediated transformation of wheat, the preferred target tissues are immature embryos. However, transformation methods relying on immature embryos require the growth of plants under controlled conditions to provide a continuous supply of good-quality target tissue. The use of mature embryos as a target tissue has the advantage of only requiring good-quality seed as the starting material. Here we describe a transformation method based on the Agrobacterium-mediated transformation of callus cultures derived from mature wheat embryos of the genotype Bobwhite S56. PMID:25300842

  10. Mapping the subgenomic RNA promoter of the Citrus leaf blotch virus coat protein gene by Agrobacterium-mediated inoculation.

    PubMed

    Renovell, Agueda; Gago, Selma; Ruiz-Ruiz, Susana; Velázquez, Karelia; Navarro, Luis; Moreno, Pedro; Vives, Mari Carmen; Guerri, José

    2010-10-25

    Citrus leaf blotch virus has a single-stranded positive-sense genomic RNA (gRNA) of 8747 nt organized in three open reading frames (ORFs). The ORF1, encoding a polyprotein involved in replication, is translated directly from the gRNA, whereas ORFs encoding the movement (MP) and coat (CP) proteins are expressed via 3' coterminal subgenomic RNAs (sgRNAs). We characterized the minimal promoter region critical for the CP-sgRNA expression in infected cells by deletion analyses using Agrobacterium-mediated infection of Nicotiana benthamiana plants. The minimal CP-sgRNA promoter was mapped between nucleotides -67 and +50 nt around the transcription start site. Surprisingly, larger deletions in the region between the CP-sgRNA transcription start site and the CP translation initiation codon resulted in increased CP-sgRNA accumulation, suggesting that this sequence could modulate the CP-sgRNA transcription. Site-specific mutational analysis of the transcription start site revealed that the +1 guanylate and the +2 adenylate are important for CP-sgRNA synthesis. PMID:20708769

  11. Agrobacterium-mediated transformation of Mexican lime (Citrus aurantifolia Swingle) using optimized systems for epicotyls and cotelydons

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Epicotyl and internodal stem segments provide the predominantly used explants for regeneration of transgenic citrus plants following co-cultivation with Agrobacterium. Previous reports using epicotyls segments from Mexican lime have shown low affinity for Agrobacterium tumefaciens infection which re...

  12. Agrobacterium tumefaciens-mediated transformation of poinsettia, Euphorbia pulcherrima, with virus-derived hairpin RNA constructs confers resistance to Poinsettia mosaic virus.

    PubMed

    Clarke, Jihong Liu; Spetz, Carl; Haugslien, Sissel; Xing, Shaochen; Dees, Merete W; Moe, Roar; Blystad, Dag-Ragnar

    2008-06-01

    Agrobacterium-mediated transformation for poinsettia (Euphorbia pulcherrima Willd. Ex Klotzsch) is reported here for the first time. Internode stem explants of poinsettia cv. Millenium were transformed by Agrobacterium tumefaciens, strain LBA 4404, harbouring virus-derived hairpin (hp) RNA gene constructs to induce RNA silencing-mediated resistance to Poinsettia mosaic virus (PnMV). Prior to transformation, an efficient somatic embryogenesis system was developed for poinsettia cv. Millenium in which about 75% of the explants produced somatic embryos. In 5 experiments utilizing 868 explants, 18 independent transgenic lines were generated. An average transformation frequency of 2.1% (range 1.2-3.5%) was revealed. Stable integration of transgenes into the poinsettia nuclear genome was confirmed by PCR and Southern blot analysis. Both single- and multiple-copy transgene integration into the poinsettia genome were found among transformants. Transgenic poinsettia plants showing resistance to mechanical inoculation of PnMV were detected by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Northern blot analysis of low molecular weight RNA revealed that transgene-derived small interfering (si) RNA molecules were detected among the poinsettia transformants prior to inoculation. The Agrobacterium-mediated transformation methodology developed in the current study should facilitate improvement of this ornamental plant with enhanced disease resistance, quality improvement and desirable colour alteration. Because poinsettia is a non-food, non-feed plant and is not propagated through sexual reproduction, this is likely to be more acceptable even in areas where genetically modified crops are currently not cultivated. PMID:18327592

  13. Agrobacterium tumefaciens-mediated transformation of poinsettia, Euphorbia pulcherrima, with virus-derived hairpin RNA constructs confers resistance to Poinsettia mosaic virus

    PubMed Central

    Spetz, Carl; Haugslien, Sissel; Xing, Shaochen; Dees, Merete W.; Moe, Roar; Blystad, Dag-Ragnar

    2008-01-01

    Agrobacterium-mediated transformation for poinsettia (Euphorbia pulcherrima Willd. Ex Klotzsch) is reported here for the first time. Internode stem explants of poinsettia cv. Millenium were transformed by Agrobacterium tumefaciens, strain LBA 4404, harbouring virus-derived hairpin (hp) RNA gene constructs to induce RNA silencing-mediated resistance to Poinsettia mosaic virus (PnMV). Prior to transformation, an efficient somatic embryogenesis system was developed for poinsettia cv. Millenium in which about 75% of the explants produced somatic embryos. In 5 experiments utilizing 868 explants, 18 independent transgenic lines were generated. An average transformation frequency of 2.1% (range 1.2–3.5%) was revealed. Stable integration of transgenes into the poinsettia nuclear genome was confirmed by PCR and Southern blot analysis. Both single- and multiple-copy transgene integration into the poinsettia genome were found among transformants. Transgenic poinsettia plants showing resistance to mechanical inoculation of PnMV were detected by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Northern blot analysis of low molecular weight RNA revealed that transgene-derived small interfering (si) RNA molecules were detected among the poinsettia transformants prior to inoculation. The Agrobacterium-mediated transformation methodology developed in the current study should facilitate improvement of this ornamental plant with enhanced disease resistance, quality improvement and desirable colour alteration. Because poinsettia is a non-food, non-feed plant and is not propagated through sexual reproduction, this is likely to be more acceptable even in areas where genetically modified crops are currently not cultivated. PMID:18327592

  14. High transformation efficiency in plum (Prunus domestica L.): a new tool for functional genomics studies in Prunus spp.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An improved Agrobacterium-mediated transformation protocol in plum (Prunus domestica L.) cv 'Bluebyrd' using hypocotyl slices as source of explants is described. The addition of 2, 4-D to the regeneration media during co-culture allowed us to increase transformation efficiency up to 10 times over p...

  15. Gateway®-compatible plant transformation vectors.

    PubMed

    Smedley, Mark A; Harwood, Wendy A

    2015-01-01

    Studies in functional genomics and crop improvement programs often rely on the introduction and expression of transgenes in plants. There are two essential components required for in planta transgene expression, a plasmid vector on which the transgene sequence is carried and a delivery system capable of transferring the vector to the target cells. Agrobacterium-mediated plant transformation and the binary plasmid vector system is the preferred method of transgene delivery. The cloning technologies used for DNA manipulation underpin many of these studies. Increased demand for efficient high-throughput transformation systems is driving forward improvements in gene cloning techniques. This chapter gives an overview of Gateway(®)-compatible binary vectors for use in Agrobacterium-mediated transformation systems. It describes a fast, efficient, and robust cloning protocol for the production of an over-expression binary vector using Gateway(®) recombinational cloning. PMID:25300827

  16. Transformation of somatic embryos of Prunus incisa ‘February Pink’ with a visible reporter gene

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An Agrobacterium-mediated transformation system was developed for the ornamental cherry species Prunus incisa. This system uses both an antibiotic resistance gene (NPTII) and a visible selectable marker, the green fluorescent protein (GFP), to select plants. Cells from leaf and root explants were tr...

  17. Production of herbicide-resistant medicinal plant Salvia miltiorrhiza transformed with the bar gene.

    PubMed

    Liu, Yu; Yang, Shi Xin; Cheng, Yan; Liu, Dong Qing; Zhang, Yong; Deng, Ke Jun; Zheng, Xue Lian

    2015-12-01

    In this study, we successfully performed Agrobacterium-mediated genetic transformation of Salvia miltiorrhiza and produced herbicide-resistant transformants. Leaf discs of S. miltiorrhiza were infected with Agrobacterium tumefaciens EHA105 harboring pCAMBIA 3301. The pCAMBIA 3301 includes an intron-containing gus reporter and a bar selection marker. To increase stable transformation efficiency, a two-step selection was employed which consists of herbicide resistance and gus expression. Here, we put more attention to the screening step of herbicide resistance. The current study provides an efficient screening system for the transformed plant of S. miltiorrhiza harboring bar gene. To determine the most suitable phosphinothricin concentration for plant selection, non-transformed leaf discs were grown on selection media containing six different phosphinothricin concentrations (0, 0.2, 0.4, 0.6, 0.8, and 1.0 mg/l). Based on the above results of non-transformed calluses, the sensitivity of phosphinothricin (0, 0.4, 0.8, 1.2, 1.6 mg/l) was tested in the screening of transgenic S. miltiorrhiza. We identified that 0.6 mg/l phosphinothricin should be suitable for selecting putatively transformed callus because non-transformed callus growth was effectively inhibited under this concentrations. When sprayed with Basta, the transgenic S. miltiorrhiza plants were tolerant to the herbicide. Hence, we report successful transformation of the bar gene conferring herbicide resistance to S. miltiorrhiza. PMID:26364310

  18. Stable transformation and expression of GhEXPA8 fiber expansin gene to improve fiber length and micronaire value in cotton

    PubMed Central

    Bajwa, Kamran S.; Shahid, Ahmad A.; Rao, Abdul Q.; Bashir, Aftab; Aftab, Asia; Husnain, Tayyab

    2015-01-01

    Cotton fiber is multigenic trait controlled by number of genes. Previous studies suggest that one of these genes may be responsible for switching cotton fiber growth on and off to influence the fiber quality produced from a cotton seed. In the present study, the Gossypium hirsutum GhEXPA8 fiber expansin gene was introduced into local cotton variety NIAB 846 by using an Agrobacterium-mediated gene transformation. The neomycin phosphotransferase (NPTII) gene was used as a selection marker for screening of putative transgenic cotton plants. Integration and expression of the fiber expansin gene in cotton plants was confirmed with molecular techniques including Southern blot analyses, real-time PCR. Cellulose assay was used for measurement of cellulose contents of transgenic cotton fiber. The data collected from 3 years of field performance of the transgenic cotton plants expressing GhEXPA8 showed that significant improvement has been made in fiber lengths and micronaire values as compared to control G. hirsutum variety NIAB 846 cotton plants. Statistical techniques were also used for analysis of fiber and agronomic characteristics. The results of this study support improvement of cotton fiber through genetic modification. PMID:26583018

  19. Long-term stability of attacin E expression in transformed apple after 12 years in the field and effect of the expression of this gene on the fruit characteristics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Transgenic trees currently are being produced by Agrobacterium-mediated transformation and biolistics. The future use of transformed trees on a commercial basis depends upon thorough evaluation of the potential environmental and public health impact of the modified plants, transgene stability over ...

  20. Genetic transformation of Diaporthe phaseolorum, an endophytic fungus found in mangrove forests, mediated by Agrobacterium tumefaciens.

    PubMed

    Sebastianes, Fernanda L S; Lacava, Paulo T; Fávaro, Léia C L; Rodrigues, Maria B C; Araújo, Welington L; Azevedo, João L; Pizzirani-Kleiner, Aline A

    2012-02-01

    We describe the genetic transformation of the mycelial tissue of Diaporthe phaseolorum, an endophytic fungus isolated from the mangrove species Laguncularia racemosa, using Agrobacterium tumefaciens-mediated transformation (ATMT). ATMT uses both the hygromycin B resistant (hph) gene and green fluorescent protein as the selection agents. The T-DNA integration into the fungal genome was assessed by both PCR and Southern blotting. All transformants examined were mitotically stable. An analysis of the T-DNA flanking sequences by thermal asymmetric interlaced PCR (TAIL-PCR) demonstrated that the disrupted genes in the transformants had similarities with conserved domains in proteins involved in antibiotic biosynthesis pathways. A library of 520 transformants was generated, and 31 of these transformants had no antibiotic activity against Staphylococcus aureus, an important human pathogen. The protocol described here, using ATMT in D. phaseolorum, will be useful for the identification and analysis of fungal genes controlling pathogenicity and antibiotic pathways. Moreover, this protocol may be used as a reference for other species in the Diaporthe genus. This is the first report to describe Agrobacterium-mediated transformation of D. phaseolorum as a tool for insertional mutagenesis. PMID:22210192

  1. Agrobacterium mediated transfer of a mutant Arabidopsis acetolactate synthase gene confers resistance to chlorsulfuron in chicory (Cichorium intybus L.).

    PubMed

    Vermeulen, A; Vaucheret, H; Pautot, V; Chupeau, Y

    1992-06-01

    Leaf discs of C. intybus were inoculated with an Agrobacterium tumefaciens strain harboring a neomycin phosphotransferase (neo) gene for kanamycin resistance and a mutant acetolactate synthase gene (csr1-1) from Arabidopsis thaliana conferring resistance to sulfonylurea herbicides. A regeneration medium was optimized which permitted an efficient shoot regeneration from leaf discs. Transgenic shoots were selected on rooting medium containing 100 mg/l kanamycin sulfate. Integration of the csr1-1 gene into genomic DNA of kanamycin resistant chicory plants was confirmed by Southern blot hybridizations. Analysis of the selfed progenies (S1 and S2) of two independent transformed clones showed that kanamycin and chlorsulfuron resistances were inherited as dominant Mendelian traits. The method described here for producing transformed plants will allow new opportunities for chicory breeding. PMID:24203132

  2. Effective elimination of chimeric tissue in transgenics for the stable genetic transformation of lesquerella fendleri

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In order to improve the potential of Lesquerella fendleri as a valuable industrial oilseed crop, a stable genetic transformation system was developed. Genetic transformation was performed by inoculating leaf segments with an Agrobacterium tumefaciens strain AGL1 carrying binary vector pCAMBIA 1301.1...

  3. Cloning and functional characterization of the promoter of PsSEOF1 gene from Pisum sativum under different stress conditions using Agrobacterium-mediated transient assay

    PubMed Central

    Srivastava, Vineet Kumar; Raikwar, Shailendra; Tuteja, Narendra

    2014-01-01

    PsSEOF1, a SEO (sieve element occlusion) gene family protein (forisome) is calcium powered motor protein and is located close to plasma membrane of sieve element. In sieve element (SE) it senses the calcium ion levels and undergoes ATP-independent conformational shifts. Forisome, meaning gate-bodies (Latin foris: wing of a gate; Greek soma: body). Recent reports show that SEO gene family protein can prevent the loss of nutrient rich photoassimilate upon wound injury. The regulation of SEO protein forisome under abiotic/ biotic stress is still unknown. The analysis of cis-regulatory element present in the upstream region is not well understood. Tissue specific promoters guarantee correct expression when it perceives particular stimuli. Here we report isolation of tissue specific promoter of PsSEOF1 was isolated by gene walking PCR from P. sativum (pea) genomic DNA library constructed by BD genome walker kit. In silico analysis revealed several putative cis element within this promoter sequence like wound response, cold, dehydration. Putative elements which might be required for its vascular tissue specificity has also been identified. The GUS activities of PsSEOF1 promoter-GUS chimeric construct in the agroinfiltrated leaves under different environmental stress abiotic and biotic like wound, cold, salt and phytohormones has shown high level of GUS activity. To identify the activity of PsSEOF1 promoter under different stress condition an Agrobacterium-mediated transient expression of tobacco plants were subjected to histochemical GUS staining. Stress-inducible nature of PsSEOF1 promoter opens possibility for the study of the PsSEOF1 gene regulation under stress condition. The isolated promoter sequence could serve as an important candidate for tissue specific promoter in genetic engineering of plant under stress conditions. PMID:25763698

  4. Development of three full-length infectious cDNA clones of distinct brassica yellows virus genotypes for agrobacterium-mediated inoculation.

    PubMed

    Zhang, Xiao-Yan; Dong, Shu-Wei; Xiang, Hai-Ying; Chen, Xiang-Ru; Li, Da-Wei; Yu, Jia-Lin; Han, Cheng-Gui

    2015-02-01

    Brassica yellows virus is a newly identified species in the genus of Polerovirus within the family Luteoviridae. Brassica yellows virus (BrYV) is prevalently distributed throughout Mainland China and South Korea, is an important virus infecting cruciferous crops. Based on six BrYV genomic sequences of isolates from oilseed rape, rutabaga, radish, and cabbage, three genotypes, BrYV-A, BrYV-B, and BrYV-C, exist, which mainly differ in the 5' terminal half of the genome. BrYV is an aphid-transmitted and phloem-limited virus. The use of infectious cDNA clones is an alternative means of infecting plants that allows reverse genetic studies to be performed. In this study, full-length cDNA clones of BrYV-A, recombinant BrYV5B3A, and BrYV-C were constructed under control of the cauliflower mosaic virus 35S promoter. An agrobacterium-mediated inoculation system of Nicotiana benthamiana was developed using these cDNA clones. Three days after infiltration with full-length BrYV cDNA clones, necrotic symptoms were observed in the inoculated leaves of N. benthamiana; however, no obvious symptoms appeared in the upper leaves. Reverse transcription-PCR (RT-PCR) and western blot detection of samples from the upper leaves showed that the maximum infection efficiency of BrYVs could reach 100%. The infectivity of the BrYV-A, BrYV-5B3A, and BrYV-C cDNA clones was further confirmed by northern hybridization. The system developed here will be useful for further studies of BrYV, such as host range, pathogenicity, viral gene functions, and plant-virus-vector interactions, and especially for discerning the differences among the three genotypes. PMID:25499296

  5. Vapor-deposited glasses of methyl-m-toluate: How uniform is stable glass transformation?

    NASA Astrophysics Data System (ADS)

    Tylinski, M.; Sepúlveda, A.; Walters, Diane M.; Chua, Y. Z.; Schick, C.; Ediger, M. D.

    2015-12-01

    AC chip nanocalorimetry is used to characterize vapor-deposited glasses of methyl-m-toluate (MMT). Physical vapor deposition can prepare MMT glasses that have lower heat capacity and significantly higher kinetic stability compared to liquid-cooled glasses. When heated, highly stable MMT glasses transform into the supercooled liquid via propagating fronts. We present the first quantitative analysis of the temporal and spatial uniformities of these transformation fronts. The front velocity varies by less than 4% over the duration of the transformation. For films 280 nm thick, the transformation rates at different spatial positions in the film differ by about 25%; this quantity may be related to spatially heterogeneous dynamics in the stable glass. Our characterization of the kinetic stability of MMT stable glasses extends previous dielectric experiments and is in excellent agreement with these results.

  6. Vapor-deposited glasses of methyl-m-toluate: How uniform is stable glass transformation?

    PubMed

    Tylinski, M; Sepúlveda, A; Walters, Diane M; Chua, Y Z; Schick, C; Ediger, M D

    2015-12-28

    AC chip nanocalorimetry is used to characterize vapor-deposited glasses of methyl-m-toluate (MMT). Physical vapor deposition can prepare MMT glasses that have lower heat capacity and significantly higher kinetic stability compared to liquid-cooled glasses. When heated, highly stable MMT glasses transform into the supercooled liquid via propagating fronts. We present the first quantitative analysis of the temporal and spatial uniformities of these transformation fronts. The front velocity varies by less than 4% over the duration of the transformation. For films 280 nm thick, the transformation rates at different spatial positions in the film differ by about 25%; this quantity may be related to spatially heterogeneous dynamics in the stable glass. Our characterization of the kinetic stability of MMT stable glasses extends previous dielectric experiments and is in excellent agreement with these results. PMID:26723694

  7. Stable Glass Transformation to Supercooled Liquid via Surface-Initiated Growth Front

    SciTech Connect

    Swallen, Stephen F.; Traynor, Katherine; McMahon, Robert J.; Ediger, M. D.; Mates, Thomas E.

    2009-02-13

    Highly stable glasses of tris-naphthylbenzene transform into a liquid when annealed above the glass transition temperature T{sub g}. In contrast to the predictions of standard models, the observed transformation is spatially inhomogeneous. Secondary ion mass spectrometry experiments on isotopically labeled multilayer films show that the liquid grows into the stable glass with sharp growth fronts initiated at the free surface and at the interface with the substrate. For the free surface, the growth velocity is constant in time and has the same temperature dependence as self-diffusion in the equilibrium supercooled liquid. These stable glasses are packed so efficiently that surfaces and interfaces are required to initiate the transformation to the liquid even well above T{sub g}.

  8. Stable nuclear transformation of Chlamydomonas reinhardtii by using a C. reinhardtii gene as the selectable marker.

    PubMed Central

    Mayfield, S P; Kindle, K L

    1990-01-01

    We have developed a stable nuclear transformation system for the unicellular green alga Chlamydomonas reinhardtii. Transformation was accomplished by introducing the cloned C. reinhardtii oxygen-evolving enhancer protein 1 (OEE1) gene into C. reinhardtii cells by bombardment with DNA-coated tungsten particles. The recipient strain was an OEE1-deficient, nonphotosynthetic, acetate-requiring mutant, which recovered photosynthetic competence after transformation, and was therefore able to grow in the absence of acetate. Analysis of several transformants indicates that transformation has proceeded via second-site integration of the cloned gene, leaving the endogenous mutant gene intact. In genetic crosses of transformants with wild type, both mutant and wild-type phenotypes were recovered, showing that the photosynthetic competence of transformants was due not to reversion of the original locus but rather to expression of the introduced gene. We suggest that the success of the present system is largely due to using a homologous C. reinhardtii gene, leading to stable maintenance and expression of the gene. Transformation with heterologous genes may be problematic because of poor expression due to an unusual codon bias in C. reinhardtii. Images PMID:2179948

  9. Phase Transformation Behavior at Low Temperature in Hydrothermal Treatment of Stable and Unstable Titania Sol

    PubMed

    So; Park; Kim; Moon

    1997-07-15

    Nanosize titania sol was prepared from titanium tetraisopropoxide (TTIP) and conditions for the formation of stable sol were identified. As the H+/TTIP mole ratio decreased and H2O/TTIP mole ratio increased, stable sol was likely to be formed. The size and crystallinity remained unchanged after hydrothermal treatment of the stable sol at between 160 and 240°C. However, hydrothermal treatment of unstable sol produced rod-like particles and crystallinity of particles was changed from anatase to rutile. This difference in phase transformation at low hydrothermal treatment temperature was likely to be caused by the fact that stable sol remained to be stable even after hydrothermal treatment, while unstable sol had a tendency to be aggregated. PMID:9268523

  10. T-DNA transfer and T-DNA integration efficiencies upon Arabidopsis thaliana root explant cocultivation and floral dip transformation.

    PubMed

    Ghedira, Rim; De Buck, Sylvie; Van Ex, Frédéric; Angenon, Geert; Depicker, Ann

    2013-12-01

    T-DNA transfer and integration frequencies during Agrobacterium-mediated root explant cocultivation and floral dip transformations of Arabidopsis thaliana were analyzed with and without selection for transformation-competent cells. Based on the presence or absence of CRE recombinase activity without or with the CRE T-DNA being integrated, transient expression versus stable transformation was differentiated. During root explant cocultivation, continuous light enhanced the number of plant cells competent for interaction with Agrobacterium and thus the number of transient gene expression events. However, in transformation competent plant cells, continuous light did not further enhance cotransfer or cointegration frequencies. Upon selection for root transformants expressing a first T-DNA, 43-69 % of these transformants showed cotransfer of another non-selected T-DNA in two different light regimes. However, integration of the non-selected cotransferred T-DNA occurred only in 19-46 % of these transformants, indicating that T-DNA integration in regenerating root cells limits the transformation frequencies. After floral dip transformation, transient T-DNA expression without integration could not be detected, while stable T-DNA transformation occurred in 0.5-1.3 % of the T1 seedlings. Upon selection for floral dip transformants with a first T-DNA, 8-34 % of the transformants showed cotransfer of the other non-selected T-DNA and in 93-100 % of them, the T-DNA was also integrated. Therefore, a productive interaction between the agrobacteria and the female gametophyte, rather than the T-DNA integration process, restricts the floral dip transformation frequencies. PMID:23975012

  11. A dark incubation period is important for Agrobacterium-mediated transformation of mature internode explants of sweet orange, grapefruit, citron, and a citrange rootstock

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Citrus has an extended juvenile phase and trees can take 2-20 years to transition to the adult reproductive phase and produce fruit. For citrus variety development this substantially prolongs the time before adult traits, such as fruit yield and quality, can be evaluated. Methods to...

  12. Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation.

    PubMed

    Qi, Jing; Dong, Zhen; Zhang, Yu-Xing

    2015-12-01

    The aim of the present study was to genetically modify plantlets of the Chinese yali pear to reduce their expression of ripening-associated 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) and therefore increase the shelf-life of the fruit. Primers were designed with selectivity for the conserved regions of published ACO gene sequences, and yali complementary DNA (cDNA) cloning was performed by reverse transcription quantitative polymerase chain reaction (PCR). The obtained cDNA fragment contained 831 base pairs, encoding 276 amino acid residues, and shared no less than 94% nucleotide sequence identity with other published ACO genes. The cDNA fragment was inversely inserted into a pBI121 expression vector, between the cauliflower mosaic virus 35S promoter and the nopaline synthase terminator, in order to construct the anti‑sense expression vector of the ACO gene; it was transfected into cultured yali plants using Agrobacterium LBA4404. Four independent transgenic lines of pear plantlets were obtained and validated by PCR analysis. A Southern blot assay revealed that there were three transgenic lines containing a single copy of exogenous gene and one line with double copies. The present study provided germplasm resources for the cultivation of novel storage varieties of pears, therefore providing a reference for further applications of anti‑sense RNA technology in the genetic improvement of pears and other fruit. PMID:26460204

  13. Analysis of sequential reactive transformations using stable isotope analysis: a theoretical assessment

    NASA Astrophysics Data System (ADS)

    Thullner, Martin; Jin, Shuang; Stadler, Susanne

    2016-04-01

    Stable isotope methods have been establish as powerful tools for the analysis of reactive transformation in the subsurface with applications ranging from the field of contaminant hydrology to biogeochemical cycling. While the link between single transformations based on stable isotope signatures and their changes is commonly well understood and allows for qualitative and - in case certain requirements are met - also quantitative analysis, the interpretation of sequential reactive transformations, e.g. decay chains, is more complicated. In the latter case isotope signature changes of individual reactive compounds are affected by more than one transformation which challenges the interpretation of these changes. In recent years, some methods have been proposed (e.g. isotope mass balance approaches) which allow at least for a qualitative or semi-quantitative estimation of sequential reactive transformation processes. However, a systematic assessment of the validity of these estimation methods is missing so far and the accuracy of these methods - in general and for specific conditions encountered in the field - is not fully validated. In this presentation we use a combination of mathematical analyses and numerical modeling to test the validity of the proposed estimation methods and to determine the limits of their applicability. Results recommend modifications of existing theoretical estimation approaches and identify sorption processes as potential limitation of their applicability.

  14. Medicago truncatula transformation using cotyledonary explants.

    PubMed

    Wright, Elane; Wang, Zeng-Yu

    2015-01-01

    Medicago truncatula has been developed into a model species for legumes. The M. truncatula genotype Jemalong A17 has been used for EST and genome sequencing. However, this genotype is difficult to regenerate from callus cultures. By using cotyledons as explants for Agrobacterium infection and direct shoot formation, this protocol allows for rapid production of transgenic plants from Jemalong A17 and other genotypes. Transgenic plants can be regenerated and established in the greenhouse in only 3-4 months after Agrobacterium-mediated transformation. Transformation frequency was in the range of 5-12 %. PMID:25300829

  15. Recent advances in plant transformation.

    PubMed

    Barampuram, Shyamkumar; Zhang, Zhanyuan J

    2011-01-01

    Plant genetic engineering has become one of the most important molecular tools in the modern molecular breeding of crops. Over the last decade, significant progress has been made in the development of new and efficient transformation methods in plants. Despite a variety of available DNA delivery methods, Agrobacterium- and biolistic-mediated transformation remain the two predominantly employed approaches. In particular, progress in Agrobacterium-mediated transformation of cereals and other recalcitrant dicot species has been quite remarkable. In the meantime, other transgenic-enabling technologies have emerged, including generation of marker-free transgenics, gene targeting, and chromosomal engineering. Although transformation of some plant species or elite germplasm remains a challenge, further advancement in transformation technology is expected because the mechanisms of governing the regeneration and transformation processes are now better understood and are being creatively applied to designing improved transformation methods or to developing new enabling technologies. PMID:21181522

  16. Transformation of radish (Raphanus sativus L.) via sonication and vacuum infiltration of germinated seeds with Agrobacterium harboring a group 3 LEA gene from B. napus.

    PubMed

    Park, Byong-Jin; Liu, Zaochang; Kanno, Akira; Kameya, Toshiaki

    2005-10-01

    A protocol for producing transgenic radish (Raphanus sativus) was obtained by using both ultrasonic and vacuum infiltration assisted, Agrobacterium-mediated transformation. The Agrobacterium strain LBA4404 contained the binary vector pBI121-LEA (late embyogenesis abundant), which carried a Group 3 LEA gene, from Brassica napus. Among six combinations, Agrobacterium-mediated transformation assisted by a combination of 5-min sonication with 5-min vacuum infiltration resulted in the highest transformation frequency. The existence, integration and expression of transferred LEA gene in transgenic T(1) plants were confirmed by PCR, genomic Southern and Western blot analysis. Transgenic radish demonstrated better growth performance than non-transformed control plants under osmotic and salt stress conditions. Accumulation of Group 3 LEA protein in the vegetative tissue of transgenic radish conferred increased tolerance to water deficit and salt stress. PMID:15843933

  17. Development of a transgenic hairy root system in jute (Corchorus capsularis L.) with gusA reporter gene through Agrobacterium rhizogenes mediated co-transformation.

    PubMed

    Chattopadhyay, Tirthartha; Roy, Sheuli; Mitra, Adinpunya; Maiti, Mrinal K

    2011-04-01

    Transgenic hairy root system is important in several recalcitrant plants, where Agrobacterium tumefaciens-mediated plant transformation and generation of transgenic plants are problematic. Jute (Corchorus spp.), the major fibre crop in Indian subcontinent, is one of those recalcitrant plants where in vitro tissue culture has provided a little success, and hence, Agrobacterium-mediated genetic transformation remains to be a challenging proposition in this crop. In the present work, a system of transgenic hairy roots in Corchorus capsularis L. has been developed through genetic transformation by Agrobacterium rhizogenes harbouring two plasmids, i.e. the natural Ri plasmid and a recombinant binary vector derived from the disarmed Ti plasmid of A. tumefaciens. Our findings indicate that the system is relatively easy to establish and reproducible. Molecular analysis of the independent lines of transgenic hairy roots revealed the transfer of relevant transgenes from both the T-DNA parts into the plant genome, indicating the co-transformation nature of the event. High level expression and activity of the gusA reporter gene advocate that the transgenic hairy root system, thus developed, could be applicable as gene expression system in general and for root functional genomics in particular. Furthermore, these transgenic hairy roots can be used in future as explants for plantlet regeneration to obtain stable transgenic jute plants. PMID:21153028

  18. Stable isotope fractionation to investigate natural transformation mechanisms of organic contaminants: principles, prospects and limitations.

    PubMed

    Elsner, Martin

    2010-11-01

    Gas chromatography-isotope ratio mass spectrometry (GC-IRMS) has made it possible to analyze natural stable isotope ratios (e.g., (13)C/(12)C, (15)N/(14)N, (2)H/(1)H) of individual organic contaminants in environmental samples. They may be used as fingerprints to infer contamination sources, and may demonstrate, and even quantify, the occurrence of natural contaminant transformation by the enrichment of heavy isotopes that arises from degradation-induced isotope fractionation. This review highlights an additional powerful feature of stable isotope fractionation: the study of environmental transformation mechanisms. Isotope effects reflect the energy difference of isotopologues (i.e., molecules carrying a light versus a heavy isotope in a particular molecular position) when moving from reactant to transition state. Measuring isotope fractionation, therefore, essentially allows a glimpse at transition states! It is shown how such position-specific isotope effects are "diluted out" in the compound average measured by GC-IRMS, and how a careful evaluation in mechanistic scenarios and by dual isotope plots can recover the underlying mechanistic information. The mathematical framework for multistep isotope fractionation in environmental transformations is reviewed. Case studies demonstrate how isotope fractionation changes in the presence of mass transfer, enzymatic commitment to catalysis, multiple chemical reaction steps or limited bioavailability, and how this gives information about the individual process steps. Finally, it is discussed how isotope ratios of individual products evolve in sequential or parallel transformations, and what mechanistic insight they contain. A concluding session gives an outlook on current developments, future research directions and the potential for bridging the gap between laboratory and real world systems. PMID:21038038

  19. Stable transformation of the yellow fever mosquito, Aedes aegypti, with the Hermes element from the housefly

    PubMed Central

    Jasinskiene, Nijole; Coates, Craig J.; Benedict, Mark Q.; Cornel, Anthony J.; Rafferty, Cristina Salazar; James, Anthony A.; Collins, Frank H.

    1998-01-01

    The mosquito Aedes aegypti is the world’s most important vector of yellow fever and dengue viruses. Work is currently in progress to control the transmission of these viruses by genetically altering the capacity of wild Ae. aegypti populations to support virus replication. The germ-line transformation system reported here constitutes a major advance toward the implementation of this control strategy. A modified Hermes transposon carrying a 4.7-kb fragment of genomic DNA that includes a wild-type allele of the Drosophila melanogaster cinnabar (cn) gene was used to transform a white-eyed recipient strain of Ae. aegypti. Microinjection of preblastoderm mosquito embryos with this construct resulted in 50% of the emergent G0 adults showing some color in their eyes. Three transformed families were recovered, each resulting from an independent insertion event of the cn+-carrying transposon. The cn+ gene functioned as a semidominant transgene and segregated in Mendelian ratios. Hermes shows great promise as a vector for efficient, heritable, and stable transformation of this important mosquito vector species. PMID:9520437

  20. Stable transformation of the yellow fever mosquito, Aedes aegypti, with the Hermes element from the housefly.

    PubMed

    Jasinskiene, N; Coates, C J; Benedict, M Q; Cornel, A J; Rafferty, C S; James, A A; Collins, F H

    1998-03-31

    The mosquito Aedes aegypti is the world's most important vector of yellow fever and dengue viruses. Work is currently in progress to control the transmission of these viruses by genetically altering the capacity of wild Ae. aegypti populations to support virus replication. The germ-line transformation system reported here constitutes a major advance toward the implementation of this control strategy. A modified Hermes transposon carrying a 4.7-kb fragment of genomic DNA that includes a wild-type allele of the Drosophila melanogaster cinnabar (cn) gene was used to transform a white-eyed recipient strain of Ae. aegypti. Microinjection of preblastoderm mosquito embryos with this construct resulted in 50% of the emergent G0 adults showing some color in their eyes. Three transformed families were recovered, each resulting from an independent insertion event of the cn+-carrying transposon. The cn+ gene functioned as a semidominant transgene and segregated in Mendelian ratios. Hermes shows great promise as a vector for efficient, heritable, and stable transformation of this important mosquito vector species. PMID:9520437

  1. Stable transformation of an episomal protein-tagging shuttle vector in the piscine diplomonad Spironucleus vortens

    PubMed Central

    Dawson, Scott C; Pham, Jonathan K; House, Susan A; Slawson, Elizabeth E; Cronembold, Daniela; Cande, W Zacheus

    2008-01-01

    Background Diplomonads are common free-living inhabitants of anoxic aquatic environments and are also found as intestinal commensals or parasites of a wide variety of animals. Spironucleus vortens is a putatively commensal diplomonad of angelfish that grows to high cell densities in axenic culture. Genomic sequencing of S. vortens is in progress, yet little information is available regarding molecular and cellular aspects of S. vortens biology beyond descriptive ultrastructural studies. To facilitate the development of S. vortens as an additional diplomonad experimental model, we have constructed and stably transformed an episomal plasmid containing an enhanced green fluorescent protein (GFP) tag, an AU1 epitope tag, and a tandem affinity purification (TAP) tag. This construct also contains selectable antibiotic resistance markers for both S. vortens and E. coli. Results Stable transformants of S. vortens grew relatively rapidly (within 7 days) after electroporation and were maintained under puromycin selection for over 6 months. We expressed the enhanced GFP variant, eGFP, under transcriptional control of the S. vortens histone H3 promoter, and visually confirmed diffuse GFP expression in over 50% of transformants. Next, we generated a histone H3::GFP fusion using the S. vortens conventional histone H3 gene and its native promoter. This construct was also highly expressed in the majority of S. vortens transformants, in which the H3::GFP fusion localized to the chromatin in both nuclei. Finally, we used fluorescence in situ hybridization (FISH) of the episomal plasmid to show that the transformed plasmid localized to only one nucleus/cell and was present at roughly 10–20 copies per nucleus. Because S. vortens grows to high densities in laboratory culture, it is a feasible diplomonad from which to purify native protein complexes. Thus, we also included a TAP tag in the plasmid constructs to permit future tagging and subsequent purification of protein complexes by

  2. Muon-Substituted Malonaldehyde: Transforming a Transition State into a Stable Structure by Isotope Substitution.

    PubMed

    Goli, Mohammad; Shahbazian, Shant

    2016-02-12

    Isotope substitutions are usually conceived to play a marginal role on the structure and bonding pattern of molecules. However, a recent study [Angew. Chem. Int. Ed. 2014, 53, 13706-13709; Angew. Chem. 2014, 126, 13925-13929] further demonstrates that upon replacing a proton with a positively charged muon, as the lightest radioisotope of hydrogen, radical changes in the nature of the structure and bonding of certain species may take place. The present report is a primary attempt to introduce another example of structural transformation on the basis of the malonaldehyde system. Accordingly, upon replacing the proton between the two oxygen atoms of malonaldehyde with the positively charged muon a serious structural transformation is observed. By using the ab initio nuclear-electronic orbital non-Born-Oppenheimer procedure, the nuclear configuration of the muon-substituted species is derived. The resulting nuclear configuration is much more similar to the transition state of the proton transfer in malonaldehyde rather than to the stable configuration of malonaldehyde. The comparison of the "atoms in molecules" (AIM) structure of the muon-substituted malonaldehyde and the AIM structure of the stable and the transition-state configurations of malonaldehyde also unequivocally demonstrates substantial similarities of the muon-substituted malonaldehyde to the transition state. PMID:26749489

  3. Identifying stable phase coupling associated with cerebral autoregulation using the synchrosqueezed cross-wavelet transform and low oscillation morlet wavelets.

    PubMed

    Addison, Paul S

    2015-08-01

    A novel method of identifying stable phase coupling behavior of two signals within the wavelet transform time-frequency plane is presented. The technique employs the cross-wavelet transform to provide a map of phase coupling followed by synchrosqueezing to collect the stable phase regime information. The resulting synchrosqueezed cross-wavelet transform method (Synchro-CrWT) is illustrated using a synthetic signal and then applied to the analysis of the relationship between biosignals used in the analysis of cerebral autoregulation function. PMID:26737649

  4. A stable isotope approach and its application for identifying nitrate source and transformation process in water.

    PubMed

    Xu, Shiguo; Kang, Pingping; Sun, Ya

    2016-01-01

    Nitrate contamination of water is a worldwide environmental problem. Recent studies have demonstrated that the nitrogen (N) and oxygen (O) isotopes of nitrate (NO3(-)) can be used to trace nitrogen dynamics including identifying nitrate sources and nitrogen transformation processes. This paper analyzes the current state of identifying nitrate sources and nitrogen transformation processes using N and O isotopes of nitrate. With regard to nitrate sources, δ(15)N-NO3(-) and δ(18)O-NO3(-) values typically vary between sources, allowing the sources to be isotopically fingerprinted. δ(15)N-NO3(-) is often effective at tracing NO(-)3 sources from areas with different land use. δ(18)O-NO3(-) is more useful to identify NO3(-) from atmospheric sources. Isotopic data can be combined with statistical mixing models to quantify the relative contributions of NO3(-) from multiple delineated sources. With regard to N transformation processes, N and O isotopes of nitrate can be used to decipher the degree of nitrogen transformation by such processes as nitrification, assimilation, and denitrification. In some cases, however, isotopic fractionation may alter the isotopic fingerprint associated with the delineated NO3(-) source(s). This problem may be addressed by combining the N and O isotopic data with other types of, including the concentration of selected conservative elements, e.g., chloride (Cl(-)), boron isotope (δ(11)B), and sulfur isotope (δ(35)S) data. Future studies should focus on improving stable isotope mixing models and furthering our understanding of isotopic fractionation by conducting laboratory and field experiments in different environments. PMID:26541149

  5. Efficient transformation and artificial miRNA gene silencing in Lemna minor.

    PubMed

    Cantó-Pastor, A; Mollá-Morales, A; Ernst, E; Dahl, W; Zhai, J; Yan, Y; Meyers, B C; Shanklin, J; Martienssen, R

    2015-01-01

    Despite rapid doubling time, simple architecture and ease of metabolic labelling, a lack of genetic tools in the Lemnaceae (duckweed) has impeded the full implementation of this organism as a model for biological research. Here, we present technologies to facilitate high-throughput genetic studies in duckweed. We developed a fast and efficient method for producing Lemna minor stable transgenic fronds via Agrobacterium-mediated transformation and regeneration from tissue culture. Additionally, we engineered an artificial microRNA (amiRNA) gene silencing system. We identified a Lemna gibba endogenous miR166 precursor and used it as a backbone to produce amiRNAs. As a proof of concept we induced the silencing of CH42, a magnesium chelatase subunit, using our amiRNA platform. Expression of CH42 in transgenic L. minor fronds was significantly reduced, which resulted in reduction of chlorophyll pigmentation. The techniques presented here will enable tackling future challenges in the biology and biotechnology of Lemnaceae. PMID:24989135

  6. Unconditionally Stable Bilinear Transformation FDTD Algorithm for Modeling Double-Negative Meta-material Electromagnetic Problems

    NASA Astrophysics Data System (ADS)

    Ramadan, Omar Salameh

    2010-03-01

    Accurate and unconditionally stable finite difference time domain (FDTD) algorithm is presented for modeling electromagnetic wave propagation in double-negative (DNG) meta-material domains. The proposed algorithm is based on incorporating the Bilinear transformation technique into the FDTD implementations of Maxwell’s equations. The stability of the proposed approach is studied by combining the von Neumann method with the Routh-Huwitz criterion and it has been observed that the proposed algorithm is free from the Courant-Friedrichs-Lewy (CFL) stability limit of the conventional FDTD scheme. Furthermore, the proposed algorithm is incorporated with the split-step FDTD scheme to model two-dimensional problems. Numerical examples carried out in one and two dimensional domains are included to show the validity of the proposed algorithm.

  7. Stable reduced-order models of generalized dynamical systems using coordinate-transformed Arnoldi algorithms

    SciTech Connect

    Silveira, L.M.; Kamon, M.; Elfadel, I.; White, J.

    1996-12-31

    Model order reduction based on Krylov subspace iterative methods has recently emerged as a major tool for compressing the number of states in linear models used for simulating very large physical systems (VLSI circuits, electromagnetic interactions). There are currently two main methods for accomplishing such a compression: one is based on the nonsymmetric look-ahead Lanczos algorithm that gives a numerically stable procedure for finding Pade approximations, while the other is based on a less well characterized Arnoldi algorithm. In this paper, we show that for certain classes of generalized state-space systems, the reduced-order models produced by a coordinate-transformed Arnoldi algorithm inherit the stability of the original system. Complete Proofs of our results will be given in the final paper.

  8. Stable recombinase-mediated cassette exchange in Arabidopsis using Agrobacterium tumefaciens.

    PubMed

    Louwerse, Jeanine D; van Lier, Miranda C M; van der Steen, Dirk M; de Vlaam, Clementine M T; Hooykaas, Paul J J; Vergunst, Annette C

    2007-12-01

    Site-specific integration is an attractive method for the improvement of current transformation technologies aimed at the production of stable transgenic plants. Here, we present a Cre-based targeting strategy in Arabidopsis (Arabidopsis thaliana) using recombinase-mediated cassette exchange (RMCE) of transferred DNA (T-DNA) delivered by Agrobacterium tumefaciens. The rationale for effective RMCE is the precise exchange of a genomic and a replacement cassette both flanked by two heterospecific lox sites that are incompatible with each other to prevent unwanted cassette deletion. We designed a strategy in which the coding region of a loxP/lox5171-flanked bialaphos resistance (bar) gene is exchanged for a loxP/lox5171-flanked T-DNA replacement cassette containing the neomycin phosphotransferase (nptII) coding region via loxP/loxP and lox5171/lox5171 directed recombination. The bar gene is driven by the strong 35S promoter, which is located outside the target cassette. This placement ensures preferential selection of RMCE events and not random integration events by expression of nptII from this same promoter. Using root transformation, during which Cre was provided on a cotransformed T-DNA, 50 kanamycin-resistant calli were selected. Forty-four percent contained a correctly exchanged cassette based on PCR analysis, indicating the stringency of the selection system. This was confirmed for the offspring of five analyzed events by Southern-blot analysis. In four of the five analyzed RMCE events, there were no additional T-DNA insertions or they easily segregated, resulting in high-efficiency single-copy RMCE events. Our approach enables simple and efficient selection of targeting events using the advantages of Agrobacterium-mediated transformation. PMID:17921337

  9. Stable Recombinase-Mediated Cassette Exchange in Arabidopsis Using Agrobacterium tumefaciens1

    PubMed Central

    Louwerse, Jeanine D.; van Lier, Miranda C.M.; van der Steen, Dirk M.; de Vlaam, Clementine M.T.; Hooykaas, Paul J.J.; Vergunst, Annette C.

    2007-01-01

    Site-specific integration is an attractive method for the improvement of current transformation technologies aimed at the production of stable transgenic plants. Here, we present a Cre-based targeting strategy in Arabidopsis (Arabidopsis thaliana) using recombinase-mediated cassette exchange (RMCE) of transferred DNA (T-DNA) delivered by Agrobacterium tumefaciens. The rationale for effective RMCE is the precise exchange of a genomic and a replacement cassette both flanked by two heterospecific lox sites that are incompatible with each other to prevent unwanted cassette deletion. We designed a strategy in which the coding region of a loxP/lox5171-flanked bialaphos resistance (bar) gene is exchanged for a loxP/lox5171-flanked T-DNA replacement cassette containing the neomycin phosphotransferase (nptII) coding region via loxP/loxP and lox5171/lox5171 directed recombination. The bar gene is driven by the strong 35S promoter, which is located outside the target cassette. This placement ensures preferential selection of RMCE events and not random integration events by expression of nptII from this same promoter. Using root transformation, during which Cre was provided on a cotransformed T-DNA, 50 kanamycin-resistant calli were selected. Forty-four percent contained a correctly exchanged cassette based on PCR analysis, indicating the stringency of the selection system. This was confirmed for the offspring of five analyzed events by Southern-blot analysis. In four of the five analyzed RMCE events, there were no additional T-DNA insertions or they easily segregated, resulting in high-efficiency single-copy RMCE events. Our approach enables simple and efficient selection of targeting events using the advantages of Agrobacterium-mediated transformation. PMID:17921337

  10. Experimental research on stable fretting wear of stainless steel wires in transformable component

    NASA Astrophysics Data System (ADS)

    Dong, Xiu-ping; Liu, Guo-quan; Zhang, Li; Bai, Hong-bai; Yang, Jian-chun

    2009-07-01

    Cool-drawn 1Cr18Ni9 stainless steel wires of φ 0.1~0.5 mm can be woven and punched to prepare transformable component which has loose, reticulate structures. When it is uploaded with vibrating force, the displacement will cause intense frictions between wires' surfaces which will dissipate abundant energy and thus it can serve as dampers like natural rubbers. Since such new type of material has double characteristics of both rubbers and metals, it is commonly called "Metal Rubber". There is certain amount of contact point/surface on wires in the transformable component and the displacements between wires are at micron levels. Experiments showed that wear course of 'fretting cell' could be plotted as four phases: polish, adherence, forming of the third bed and stabilization. The stabilization phase, in which the friction coefficients are comparatively stable, dominates the whole course. Based on data of Metal Rubber vibration fatigue experiment, φ 0.3 mm cool-drawn 1Cr18Ni9 stainless steel wires' dry fretting experiments at 10 N load are made on SRV high temperature wear tester, friction coefficients are collected and fret traces are studied by laser scanning confocal microscope (LSCM). Results indicate that wire's stabilization wear phase is the circulation process of grindings' forming, concentrating to blocks of φ 20 μm, busting and discharging. Deformation induced martensite transit in wire's cool drawing has significant effects on grinding blocks' bursting performances.

  11. Compound specific stable isotope analysis (CSIA) to characterize transformation mechanisms of α-hexachlorocyclohexane.

    PubMed

    Zhang, Ning; Bashir, Safdar; Qin, Jinyi; Schindelka, Janine; Fischer, Anko; Nijenhuis, Ivonne; Herrmann, Hartmut; Wick, Lukas Y; Richnow, Hans H

    2014-09-15

    A systematic investigation of environmentally relevant transformation processes of alpha-hexachlorocyclohexane (α-HCH) was performed in order to explore the potential of compound specific stable isotope analysis (CSIA) to characterize reaction mechanisms. The carbon isotope enrichment factors (ɛC) for the chemical transformations of α-HCH via direct photolysis, indirect photolysis (UV/H2O2), hydrolysis, electro-reduction or reduction by Fe(0) were quantified and compared to those previously published for biodegradation. Hydrogen abstraction by hydroxyl radicals generated by UV/H2O2 led to ɛC of -1.9 ± 0.2 ‰ with an apparent kinetic carbon isotope effect (AKIEC) of 1.012 ± 0.001. Dehydrochlorination by alkaline hydrolysis yielded ɛC of -7.6 ± 0.4 ‰ with AKIEC of 1.048 ± 0.003. Dechlorination either by homolytic bond cleavage in direct photolysis (ɛC=-2.8 ± 0.2 ‰) or single-electron transfer in electro-reduction (ɛC=-3.8 ± 0.4 ‰) corresponded to AKIEC of 1.017 ± 0.001 and 1.023 ± 0.003, respectively. Dichloroelimination catalyzed by Fe(0) via two-electron transfers resulted in ɛC of -4.9 ± 0.1 ‰. AKIEC values assuming either a concerted or a stepwise mechanism were 1.030 ± 0.0006 and 1.015 ± 0.0003, respectively. Contrary to biodegradation, no enantioselectivity of α-HCH was observed in chemical reactions, which might be used to discriminate chemical and biological in situ transformations. PMID:25238192

  12. Stable Transformation of Ferns Using Spores as Targets: Pteris vittata and Ceratopteris thalictroides1[W][OPEN

    PubMed Central

    Muthukumar, Balasubramaniam; Joyce, Blake L.; Elless, Mark P.; Stewart, C. Neal

    2013-01-01

    Ferns (Pteridophyta) are very important members of the plant kingdom that lag behind other taxa with regards to our understanding of their genetics, genomics, and molecular biology. We report here, to our knowledge, the first instance of stable transformation of fern with recovery of transgenic sporophytes. Spores of the arsenic hyperaccumulating fern Pteris vittata and tetraploid ‘C-fern Express’ (Ceratopteris thalictroides) were stably transformed by Agrobacterium tumefaciens with constructs containing the P. vittata actin promoter driving a GUSPlus reporter gene. Reporter gene expression assays were performed on multiple tissues and growth stages of gametophytes and sporophytes. Southern-blot analysis confirmed stable transgene integration in recovered sporophytes and also confirmed that no plasmid from A. tumefaciens was present in the sporophyte tissues. We recovered seven independent transformants of P. vittata and four independent C. thalictroides transgenics. Inheritance analyses using β-glucuronidase (GUS) histochemical staining revealed that the GUS transgene was stably expressed in second generation C. thalictroides sporophytic tissues. In an independent experiment, the gusA gene that was driven by the 2× Cauliflower mosaic virus 35S promoter was bombarded into P. vittata spores using biolistics, in which putatively stable transgenic gametophytes were recovered. Transformation procedures required no tissue culture or selectable marker genes. However, we did attempt to use hygromycin selection, which was ineffective for recovering transgenic ferns. This simple stable transformation method should help facilitate functional genomics studies in ferns. PMID:23933990

  13. Nitrogen stable isotopes in streams: effects of agricultural sources and transformations.

    PubMed

    Diebel, Matthew W; Vander Zanden, M Jake

    2009-07-01

    The nitrogen stable isotope ratio of biological tissue has been proposed as an indicator of anthropogenic N inputs to aquatic ecosystems, but overlap in the isotopic signatures of various N sources and transformations make definitive attribution of processes difficult. We collected primary consumer invertebrates from streams in agricultural settings in Wisconsin, U.S.A., to evaluate the relative influence of animal manure, inorganic fertilizer, and denitrification on biotic delta15N. Variance in biotic delta15N was explained by inorganic fertilizer inputs and the percentage of wetland land cover in the watershed, but not by animal manure inputs. These results suggest that denitrification of inorganic fertilizer is the primary driver of delta15N variability among the study sites. Comparison with previously collected stream water NO3-N concentrations at the same sites supports the role of denitrification; for a given N application rate, streams with high biotic delta15N had low NO3-N concentrations. The lack of a manure signal in biotic delta15N may be due its high ammonia content, which can be dispersed outside the range of its application by volatilization. Based on our findings and on agricultural census data for the entire United States, inorganic fertilizer is more likely than manure to drive variability in biotic delta15N and to cause excessive nitrogen concentrations in streams. PMID:19688921

  14. Evaluation of Selectable Markers for Obtaining Stable Transformants in the Gramineae 1

    PubMed Central

    Hauptmann, R. M.; Vasil, V.; Ozias-Akins, P.; Tabaeizadeh, Z.; Rogers, S. G.; Fraley, R. T.; Horsch, Robert B.; Vasil, Indra K.

    1988-01-01

    Cell suspension cultures of Triticum monococcum, Panicum maximum, Saccharum officinarum, Pennisetum americanum, and a double cross trispecific hybrid between Pennisetum americanum, P. purpureum, and P. squamulatum were tested for resistance to kanamycin, hygromycin, and methotrexate for use in transformation studies. All cultures showed high natural levels of resistance to kanamycin, in excess of 800 milligrams per liter, and variable levels of resistance to hygromycin. Methotrexate was a potent growth inhibitor at low concentrations with all species. Kanamycin and hygromycin were growth inhibitory only if added early (within 5 days after protoplast isolation and culture). Protoplasts of T. monococcum, P. maximum, S. officinarum, and the tri-specific hybrid were electroporated with plasmid DNA containing hygromycin (pMON410), kanamycin (pMON273), or methotrexate (pMON806) resistance genes. Resistant colonies were obtained at low frequencies (1 × 10−5 to 2 × 10−6) when selected under conditions which were growth inhibitory to protoplasts electroporated without DNA. Southern blot hybridization confirmed stable integration of plasmid DNA into T. monococcum using hygromycin vectors and P. maximum using the methotrexate vector with 1 to 10 copies integrated per haploid genome. Images Fig. 2 Fig. 3 PMID:16665953

  15. Agrobacterium-mediated disruption of a nonribosomal peptide synthetase gene in the invertebrate pathogen Metarhizium anisopliae reveals a peptide spore factor

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Numerous secondary metabolites have been isolated from the insect pathogenic fungus Metarhizium anisopliae, but the roles of these compounds as virulence factors in disease development are poorly understood. We targeted for disruption by Agrobacterium tumefaciens-mediated transformation a putative n...

  16. Evidence for stable transformation of wheat by floraldip in Agrobacterium tumefaciens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hexaploid wheat is one of the world’s most important staple crops but genetic transformation is still challenging. We have developed a floral transformation protocol that does not utilize tissue culture. Three T-DNA wheat transformants have been produced in the germplasm line, Crocus, using this p...

  17. Optimization of in vitro regeneration and Agrobacterium tumefaciens-mediated transformation with heat-resistant cDNA in Brassica oleracea subsp. italica cv. Green Marvel.

    PubMed

    Ravanfar, Seyed Ali; Aziz, Maheran Abdul; Saud, Halimi Mohd; Abdullah, Janna Ong

    2015-11-01

    An efficient system for shoot regeneration and Agrobacterium tumefaciens-mediated transformation of Brassica oleracea cv. Green Marvel cultivar is described. This study focuses on developing shoot regeneration from hypocotyl explants of broccoli cv. Green Marvel using thidiazuron (TDZ), zeatin, and kinetin, the optimization of factors affecting Agrobacterium-mediated transformation of the hypocotyl explants with heat-resistant cDNA, followed by the confirmation of transgenicity of the regenerants. High shoot regeneration was observed in 0.05-0.1 mg dm(-3) TDZ. TDZ at 0.1 mg dm(-3) produced among the highest percentage of shoot regeneration (96.67 %) and mean number of shoot formation (6.17). The highest percentage (13.33 %) and mean number (0.17) of putative transformant production were on hypocotyl explants subjected to preculture on shoot regeneration medium (SRM) with 200 µM acetosyringone. On optimization of bacterial density and inoculation time, the highest percentage and mean number of putative transformant production were on hypocotyl explants inoculated with a bacterial dilution of 1:5 for 30 min. Polymerase chain reaction (PCR) assay indicated a transformation efficiency of 8.33 %. The luciferase assay showed stable integration of the Arabidopsis thaliana HSP101 (AtHSP101) cDNA in the transgenic broccoli regenerants. Three out of five transgenic lines confirmed through PCR showed positive hybridization bands of the AtHSP101 cDNA through Southern blot analysis. The presence of AtHSP101 transcripts in the three transgenic broccoli lines indicated by reverse transcription-PCR (RT-PCR) confirmed the expression of the gene. In conclusion, an improved regeneration system has been established from hypocotyl explants of broccoli followed by successful transformation with AtHSP101 for resistance to high temperature. PMID:25986972

  18. Arsenicals produce stable progressive changes in DNA methylation patterns that are linked to malignant transformation of immortalized urothelial cells

    SciTech Connect

    Jensen, Taylor J.; Novak, Petr; Wnek, Shawn M.; Gandolfi, A. Jay; Futscher, Bernard W.

    2009-12-01

    Aberrant DNA methylation participates in carcinogenesis and is a molecular hallmark of a tumor cell. Tumor cells generally exhibit a redistribution of DNA methylation resulting in global hypomethylation with regional hypermethylation; however, the speed in which these changes emerge has not been fully elucidated and may depend on the temporal location of the cell in the path from normal, finite lifespan to malignant transformation. We used a model of arsenical-induced malignant transformation of immortalized human urothelial cells and DNA methylation microarrays to examine the extent and temporal nature of changes in DNA methylation that occur during the transition from immortal to malignantly transformed. Our data presented herein suggest that during arsenical-induced malignant transformation, aberrant DNA methylation occurs non-randomly, progresses gradually at hundreds of gene promoters, and alters expression of the associated gene, and these changes are coincident with the acquisition of malignant properties, such as anchorage independent growth and tumor formation in immunocompromised mice. The DNA methylation changes appear stable, since malignantly transformed cells removed from the transforming arsenical exhibited no reversion in DNA methylation levels, associated gene expression, or malignant phenotype. These data suggest that arsenicals act as epimutagens and directly link their ability to induce malignant transformation to their actions on the epigenome.

  19. Agrobacterium-Mediated Disruption of a Nonribosomal Peptide Synthetase Gene in the Invertebrate Pathogen Metarhizium anisopliae Reveals a Peptide Spore Factor▿ †

    PubMed Central

    Moon, Yong-Sun; Donzelli, Bruno G. G.; Krasnoff, Stuart B.; McLane, Heather; Griggs, Mike H.; Cooke, Peter; Vandenberg, John D.; Gibson, Donna M.; Churchill, Alice C. L.

    2008-01-01

    Numerous secondary metabolites have been isolated from the insect pathogenic fungus Metarhizium anisopliae, but the roles of these compounds as virulence factors in disease development are poorly understood. We targeted for disruption by Agrobacterium tumefaciens-mediated transformation a putative nonribosomal peptide synthetase (NPS) gene, MaNPS1. Four of six gene disruption mutants identified were examined further. Chemical analyses showed the presence of serinocyclins, cyclic heptapeptides, in the extracts of conidia of control strains, whereas the compounds were undetectable in ΔManps1 mutants treated identically or in other developmental stages, suggesting that MaNPS1 encodes a serinocyclin synthetase. Production of the cyclic depsipeptide destruxins, M. anisopliae metabolites also predicted to be synthesized by an NPS, was similar in ΔManps1 mutant and control strains, indicating that MaNPS1 does not contribute to destruxin biosynthesis. Surprisingly, a MaNPS1 fragment detected DNA polymorphisms that correlated with relative destruxin levels produced in vitro, and MaNPS1 was expressed concurrently with in vitro destruxin production. ΔManps1 mutants exhibited in vitro development and responses to external stresses comparable to control strains. No detectable differences in pathogenicity of the ΔManps1 mutants were observed in bioassays against beet armyworm and Colorado potato beetle in comparison to control strains. This is the first report of targeted disruption of a secondary metabolite gene in M. anisopliae, which revealed a novel cyclic peptide spore factor. PMID:18502925

  20. Advances in Agrobacterium tumefaciens-mediated genetic transformation of graminaceous crops.

    PubMed

    Singh, Roshan Kumar; Prasad, Manoj

    2016-05-01

    Steady increase in global population poses several challenges to plant science research, including demand for increased crop productivity, grain yield, nutritional quality and improved tolerance to different environmental factors. Transgene-based approaches are promising to address these challenges by transferring potential candidate genes to host organisms through different strategies. Agrobacterium-mediated gene transfer is one such strategy which is well known for enabling efficient gene transfer in both monocot and dicots. Due to its versatility, this technique underwent several advancements including development of improved in vitro plant regeneration system, co-cultivation and selection methods, and use of hyper-virulent strains of Agrobacterium tumefaciens harbouring super-binary vectors. The efficiency of this method has also been enhanced by the use of acetosyringone to induce the activity of vir genes, silver nitrate to reduce the Agrobacterium-induced necrosis and cysteine to avoid callus browning during co-cultivation. In the last two decades, extensive efforts have been invested towards achieving efficient Agrobacterium-mediated transformation in cereals. Though high-efficiency transformation systems have been developed for rice and maize, comparatively lesser progress has been reported in other graminaceous crops. In this context, the present review discusses the progress made in Agrobacterium-mediated transformation system in rice, maize, wheat, barley, sorghum, sugarcane, Brachypodium, millets, bioenergy and forage and turf grasses. In addition, it also provides an overview of the genes that have been recently transferred to these graminaceous crops using Agrobacterium, bottlenecks in this technique and future possibilities for crop improvement. PMID:26660352

  1. Stable transformation of Mesembryanthemum crystallinum (L.) with Agrobacterium rhizogenes harboring the green fluorescent protein targeted to the endoplasmic reticulum.

    PubMed

    Konieczny, Robert; Obert, Bohuš; Bleho, Juraj; Novák, Ondřej; Heym, Claudia; Tuleja, Monika; Müller, Jens; Strnad, Miroslav; Menzel, Diedrik; Samaj, Jozef

    2011-05-01

    Stable transformation of Mesembryanthemum crystallinum L. (common ice plant) with a green fluorescent protein (GFP) construct targeted to the endoplasmic reticulum was obtained. Seven and fourteen days after germination seedlings were infected with Agrobacterium rhizogenes strain ARqua1 either by direct coating of the cut radicles with bacteria growing on solid medium or by immersion of the cut surface in bacterial suspension at different optical densities. Both methods of infection resulted in production of GFP-positive roots with a frequency ranging from 6 to 20% according to the age of the explants and the application procedure. The green fluorescing roots displayed the typical hairy root phenotype and were easily maintained in liquid medium without growth regulators for over 2 years. Stable expression of the transgene in the roots was confirmed by polymerase chain reaction (PCR), immunoblotting and the capacity of roots to grow and produce callus on kanamycin-enriched medium. Nineteen endogenous cytokinins were determined in transgenic and non-transformed roots. The results revealed significantly lower levels of the free bases of isopentenyladenine, dihydrozeatin, cis- and trans-zeatin, as well as a conspicuous decline in concentrations of the corresponding nucleosides and most nucleotides in transgenic roots compared to the wild type. Comparison of the cytokinin profiles in transgenic and non-transformed roots suggested that transformation by A. rhizogenes disturbed cytokinin metabolism during the early steps of biosynthesis. Calli obtained from transformed roots were GFP-positive and remained non-regenerative or displayed high rhizogenic potential depending on the auxin/cytokinin ratio in the medium. Calli and callus-derived roots showed a strong GFP signal for over 2 years. PMID:21195506

  2. Stable transformation of Populus and incorporation of pest resistance by electric discharge particle acceleration.

    PubMed

    McCown, B H; McCabe, D E; Russell, D R; Robison, D J; Barton, K A; Raffa, K F

    1991-02-01

    Three different target tissues (protoplast-derived cells, nodules, and stems) and two unrelated hybrid genotypes of Populus (P. alba x P. grandidentata 'Crandon' and P. nigra 'Betulifolia' x P. trichocarpa) have been stably transformed by electric discharge particle acceleration using a 18.7 kb plasmid containing NOS-NPT, CaMV 35S-GUS, and CaMV 35S-BT. Four transformed plants of one hybrid genotype, NC5339, containing all 3 genes were recovered and analyzed. Two expressed GUS and one was highly resistant to feeding by 2 lepidopteran pests (the forest tent caterpillar, Malacosoma disstria, and the gypsy moth, Lymantria dispar.) Pretreatment of the target tissues, fine-tuning of the bombardment parameters, and the use of a selection technique employing flooding of the target tissues were important for reliable recovery of transformed plants. PMID:24220719

  3. Stable transformation of the cotton plastid genome and maternal inheritance of transgenes

    PubMed Central

    Kumar, Shashi; Dhingra, Amit; Daniell, Henry

    2012-01-01

    Chloroplast genetic engineering overcomes concerns of gene containment, low levels of transgene expression, gene silencing, positional and pleiotropic effects or presence of vector sequences in transformed genomes. Several therapeutic proteins and agronomic traits have been highly expressed via the tobacco chloroplast genome but extending this concept to important crops has been a major challenge; lack of 100% homologous species-specific chloroplast transformation vectors containing suitable selectable markers, ability to regulate transgene expression in developing plastids and inadequate tissue culture systems via somatic embryogenesis are major challenges. We employed a ‘Double Gene/Single Selection (DGSS)’ plastid transformation vector that harbors two selectable marker genes (aphA-6 and nptII) to detoxify the same antibiotic by two enzymes, irrespective of the type of tissues or plastids; by combining this with an efficient regeneration system via somatic embryogenesis, cotton plastid transformation was achieved for the first time. The DGSS transformation vector is at least 8-fold (1 event/2.4 bombarded plates) more efficient than ‘Single Gene/Single Selection (SGSS)’ vector (aphA-6; 1 event per 20 bombarded plates). Chloroplast transgenic lines were fertile, flowered and set seeds similar to untransformed plants. Transgenes stably integrated into the cotton chloroplast genome were maternally inherited and were not transmitted via pollen when out-crossed with untransformed female plants. Cotton is one of the most important genetically modified crops ($ 120 billion US annual economy). Successful transformation of the chloroplast genome should address concerns about transgene escape, insects developing resistance, inadequate insect control and promote public acceptance of genetically modified cotton. PMID:15604738

  4. Teaching Stable Two-Mirror Resonators through the Fractional Fourier Transform

    ERIC Educational Resources Information Center

    Moreno, Ignacio; Garcia-Martinez, Pascuala; Ferreira, Carlos

    2010-01-01

    We analyse two-mirror resonators in terms of their fractional Fourier transform (FRFT) properties. We use the basic ABCD ray transfer matrix method to show how the resonator can be regarded as the cascade of two propagation-lens-propagation FRFT systems. Then, we present a connection between the geometric properties of the resonator (the g…

  5. Genetic transformation of Vitis vinifera via organogenesis

    PubMed Central

    Mezzetti, Bruno; Pandolfini, Tiziana; Navacchi, Oriano; Landi, Lucia

    2002-01-01

    Background Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. The current methods for the production of transgenic grape plants are based on Agrobacterium-mediated transformation followed by regeneration from embryogenic callus. However, grape embryogenic calli are laborious to establish and the phenotype of the regenerated plants can be altered. Results Transgenic grape plants (V. vinifera, table-grape cultivars Silcora and Thompson Seedless) were produced using a method based on regeneration via organogenesis. In vitro proliferating shoots were cultured in the presence of increasing concentrations of N6-benzyl adenine. The apical dome of the shoot was removed at each transplantation which, after three months, produced meristematic bulk tissue characterized by a strong capacity to differentiate adventitious shoots. Slices prepared from the meristematic bulk were used for Agrobacterium-mediated transformation of grape plants with the gene DefH9-iaaM. After rooting on kanamycin containing media and greenhouse acclimatization, transgenic plants were transferred to the field. At the end of the first year of field cultivation, DefH9-iaaM grape plants were phenotypically homogeneous and did not show any morphological alterations in vegetative growth. The expression of DefH9-iaaM gene was detected in transgenic flower buds of both cultivars. Conclusions The phenotypic homogeneity of the regenerated plants highlights the validity of this method for both propagation and genetic transformation of table grape cultivars. Expression of the DefH9-iaaM gene takes place in young flower buds of transgenic plants from both grape cultivars. PMID:12354328

  6. Stable yeast transformants that secrete functional. cap alpha. -amylase encoded by cloned mouse pancreatic cDNA

    SciTech Connect

    Filho, S.A.; Galembeck, E.V.; Faria, J.B.; Frascino, A.C.S.

    1986-04-01

    Mouse pancreatic ..cap alpha..-amylase complementary DNA was inserted into a yeast shuttle vector after the Saccharomyces cerevisiae MF..cap alpha..1 promoter and secretion signals coding sequences. When transformed with the recombinant plasmid, S. cerevisiae cells were able to synthesize and secrete functional ..cap alpha..-amylase, efficiently hydrolyzing starch present in the culture medium. Stable amylolytic cells were obtained from different yeast strains. This work represents a significant step towards producing yeast that can convert starchy materials directly to ethanol.

  7. Development of efficient catharanthus roseus regeneration and transformation system using agrobacterium tumefaciens and hypocotyls as explants

    PubMed Central

    2012-01-01

    overexpression of DAT increased the yield of vindoline in transgenic plants. Conclusions In the present study, we report an efficient Agrobacterium-mediated transformation system for C. roseus plants with 11% of transformation frequency. To our knowledge, this is the first report on the establishment of A. tumefaciens mediated transformation and regeneration of C. roseus. More importantly, the C. roseus transformation system developed in this work was confirmed in the successful transformation of C. roseus using a key gene DAT involved in TIAs biosynthetic pathway resulting in the higher accumulation of vindoline in transgenic plants. PMID:22748182

  8. Stress-induced phase transformation and optical coupling of silver nanoparticle superlattices into mechanically stable nanowires.

    PubMed

    Li, Binsong; Wen, Xiaodong; Li, Ruipeng; Wang, Zhongwu; Clem, Paul G; Fan, Hongyou

    2014-01-01

    One-dimensional silver materials display unique optical and electrical properties with promise as functional blocks for a new generation of nanoelectronics. To date, synthetic approaches and property engineering of silver nanowires have primarily focused on chemical methods. Here we report a simple physical method of metal nanowire synthesis, based on stress-induced phase transformation and sintering of spherical Ag nanoparticle superlattices. Two phase transformations of nanoparticles under stress have been observed at distinct length scales. First, the lattice dimensions of silver nanoparticle superlattices may be reversibly manipulated between 0-8 GPa compressive stresses to enable systematic and reversible changes in mesoscale optical coupling between silver nanoparticles. Second, stresses greater than 8 GPa induced an atomic lattice phase transformation, which induced sintering of silver nanoparticles into micron-length scale nanowires. The nanowire synthesis mechanism displays a dependence on both nanoparticle crystal surface orientation and presence of particular grain boundaries to enable nanoparticle consolidation into nanowires. PMID:24957078

  9. High efficiency transformation of banana [Musa acuminata L. cv. Matti (AA)] for enhanced tolerance to salt and drought stress through overexpression of a peanut salinity-induced pathogenesis-related class 10 protein.

    PubMed

    Rustagi, Anjana; Jain, Shalu; Kumar, Deepak; Shekhar, Shashi; Jain, Mukesh; Bhat, Vishnu; Sarin, Neera Bhalla

    2015-01-01

    Bananas and plantains (Musa spp. L.) are important subsistence crops and premium export commodity in several countries, and susceptible to a wide range of environmental and biotic stress conditions. Here, we report efficient, rapid, and reproducible Agrobacterium-mediated transformation and regeneration of an Indian niche cultivar of banana [M. acuminata cv. Matti (AA)]. Apical meristem-derived highly proliferative multiple shoot clump (MSC) explants were transformed with the Agrobacterium strain EHA105 harboring a binary vector pCAMBIA-1301 carrying hptII and uidA. Sequential agro-infiltration (10 min, 400 mmHg), infection (additional 35 min, Agrobacterium density A 600 = 0.8) and co-cultivation (18 h) regimen in 100 µM acetosyringone containing liquid medium were critical factors yielding high transformation efficiency (~81 %) corroborated by transient GUS expression assay. Stable transgenic events were recovered following two cycles of meristem initiation and selection on hygromycin containing medium. Histochemical GUS assay in several tissues of transgenic plants and molecular analyses confirmed stable integration and expression of transgene. The protocol described here allowed recovery of well-established putative transgenic plantlets in as little as 5 months. The transgenic banana plants could be readily acclimatized under greenhouse conditions, and were phenotypically similar to the wild-type untransformed control plants (WT). Transgenic plants overexpressing Salinity-Induced Pathogenesis-Related class 10 protein gene from Arachis hypogaea (AhSIPR10) in banana cv. Matti (AA) showed better photosynthetic efficiency and less membrane damage (P < 0.05) in the presence of NaCl and mannitol in comparison to WT plants suggesting the role of AhSIPR10 in better tolerance of salt stress and drought conditions. PMID:25173686

  10. Stable Plastid Transformation for High-Level Recombinant Protein Expression: Promises and Challenges

    PubMed Central

    Gao, Meili; Li, Yongfei; Xue, Xiaochang; Wang, Xianfeng; Long, Jiangang

    2012-01-01

    Plants are a promising expression system for the production of recombinant proteins. However, low protein productivity remains a major obstacle that limits extensive commercialization of whole plant and plant cell bioproduction platform. Plastid genetic engineering offers several advantages, including high levels of transgenic expression, transgenic containment via maternal inheritance, and multigene expression in a single transformation event. In recent years, the development of optimized expression strategies has given a huge boost to the exploitation of plastids in molecular farming. The driving forces behind the high expression level of plastid bioreactors include codon optimization, promoters and UTRs, genotypic modifications, endogenous enhancer and regulatory elements, posttranslational modification, and proteolysis. Exciting progress of the high expression level has been made with the plastid-based production of two particularly important classes of pharmaceuticals: vaccine antigens, therapeutic proteins, and antibiotics and enzymes. Approaches to overcome and solve the associated challenges of this culture system that include low transformation frequencies, the formation of inclusion bodies, and purification of recombinant proteins will also be discussed. PMID:23093835

  11. Stable isotope (C, O) and monovalent cation fractionation upon synthesis of carbonate-bearing hydroxyl apatite (CHAP) via calcite transformation

    NASA Astrophysics Data System (ADS)

    Böttcher, Michael E.; Schmiedinger, Iris; Wacker, Ulrike; Conrad, Anika C.; Grathoff, Georg; Schmidt, Burkhard; Bahlo, Rainer; Gehlken, Peer-L.; Fiebig, Jens

    2016-04-01

    Carbonate-bearing hydroxyl-apatite (CHAP) is of fundamental and applied interest to the (bio)geochemical, paleontological, medical and material science communities, since it forms the basic mineral phase in human and animal teeth and bones. In addition, it is found in non-biogenic phosphate deposits. The stable isotope and foreign element composition of biogenic CHAP is widely used to estimate the formation conditions. This requires careful experimental calibration under well-defined boundary conditions. Within the DFG project EXCALIBOR, synthesis of carbonate-bearing hydroxyapatite was conducted via the transformation of synthetic calcite powder in aqueous solution as a function of time, pH, and temperature using batch-type experiments. The aqueous solution was analyzed for the carbon isotope composition of dissolved inorganic carbonate (gas irmMS), the oxygen isotope composition of water (LCRDS), and the cationic composition. The solid was characterized by powder X-ray diffraction, micro Raman and FTIR spectroscopy, SEM-EDX, elemental analysis (EA, ICP-OES) and gas irmMS. Temperature was found to significantly impact the transformation rate of calcite to CHAP. Upon complete transformation, CHAP was found to contain up to 5% dwt carbonate, depending on the solution composition (e.g., pH), both incorporated on the A and B type position of the crystal lattice. The oxygen isotope fractionation between water and CHAP decreased with increasing temperature with a tentative slope shallower than those reported in the literature for apatite, calcite or aragonite. In addition, the presence of dissolved NH4+, K+ or Na+ in aqueous solution led to partial incorporation into the CHAP lattice. How these distortions of the crystal lattice may impact stable isotope discrimination is subject of future investigations.

  12. Transformation of human cathelicidin LL-37 into selective, stable, and potent antimicrobial compounds.

    PubMed

    Wang, Guangshun; Hanke, Mark L; Mishra, Biswajit; Lushnikova, Tamara; Heim, Cortney E; Chittezham Thomas, Vinai; Bayles, Kenneth W; Kielian, Tammy

    2014-09-19

    This Letter reports a family of novel antimicrobial compounds obtained by combining peptide library screening with structure-based design. Library screening led to the identification of a human LL-37 peptide resistant to chymotrypsin. This d-amino-acid-containing peptide template was active against Escherichia coli but not methicillin-resistant Staphylococcus aureus (MRSA). It possesses a unique nonclassic amphipathic structure with hydrophobic defects. By repairing the hydrophobic defects, the peptide (17BIPHE2) gained activity against the ESKAPE pathogens, including Enterococcus faecium, S. aureus, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa, and Enterobacter species. In vitro, 17BIPHE2 could disrupt bacterial membranes and bind to DNA. In vivo, the peptide prevented staphylococcal biofilm formation in a mouse model of catheter-associated infection. Meanwhile, it boosted the innate immune response to further combat the infection. Because these peptides are potent, cell-selective, and stable to several proteases, they may be utilized to combat one or more ESKAPE pathogens. PMID:25061850

  13. Stable silver isotope fractionation in the natural transformation process of silver nanoparticles

    NASA Astrophysics Data System (ADS)

    Lu, Dawei; Liu, Qian; Zhang, Tuoya; Cai, Yong; Yin, Yongguang; Jiang, Guibin

    2016-08-01

    Nanoparticles in the environment can form by natural processes or be released due to human activities. Owing to limited analytical methods, the behaviour of nanoparticles in the natural environment is poorly understood and until now they have only been described by the variations in the nanoparticle size or the concentration of the element of interest. Here we show that by using inductively coupled plasma mass spectrometry to measure silver (Ag) isotope ratios it is possible to understand the transformation processes of silver nanoparticles (AgNPs) in the environment. We found that the formation and dissolution of AgNPs under natural conditions caused significant variations in the ratio of natural Ag isotopes (107Ag and 109Ag) with an isotopic enrichment factor (ε) up to 0.86‰. Furthermore, we show that engineered AgNPs have distinctly different isotope fractionation effects to their naturally formed counterparts. Further studies will be needed to understand whether isotope analysis can be used to reveal the sources of AgNPs in the environment.

  14. Construction and Validation of a Dual-Transgene Vector System for Stable Transformation in Plants.

    PubMed

    He, Zhimin; Liu, Bin; Wang, Xu; Bian, Mingdi; He, Reqing; Yan, Jindong; Zhong, Ming; Zhao, Xiaoying; Liu, Xuanming

    2016-04-20

    In this study, we constructed dual-transgene vectors (pDT1, pDT7, and pDT7G) that simultaneously co-expressed two genes in plants. ACTIN2 and UBQ10 promoters were used to control the expression of these two genes. The 4×Myc, 3×HA, and 3×Flag reporter genes allowed for the convenient identification of a tunable co-expression system in plants, whereas the dexamethasone (Dex) inducible reporter gene C-terminus of the glucocorticoid receptor (cGR) provided Dex-dependent translocation of the fusion gene between the nucleus and cytoplasm. The function of pDT vectors was validated using four pairwise genes in Nicotiana benthamiana or Arabidopsis thaliana. The co-expression efficiency of two genes from the pDT1 and pDT7G vectors was 35% and 42%, respectively, which ensured the generation of sufficient transgenic materials. These pDT vectors are simple, reliable, efficient, and time-saving tools for the co-expression of two genes through a single transformation event and can be used in the study of protein-protein interactions or multi-component complexes. PMID:27157807

  15. Recent advances in soybean transformation and their application to molecular breeding and genomic analysis

    PubMed Central

    Yamada, Tetsuya; Takagi, Kyoko; Ishimoto, Masao

    2012-01-01

    Herbicide-resistant transgenic soybean plants hold a leading market share in the USA and other countries, but soybean has been regarded as recalcitrant to transformation for many years. The cumulative and, at times, exponential advances in genetic manipulation have made possible further choices for soybean transformation. The most widely and routinely used transformation systems are cotyledonary node–Agrobacterium-mediated transformation and somatic embryo–particle-bombardment-mediated transformation. These ready systems enable us to improve seed qualities and agronomic characteristics by transgenic approaches. In addition, with the accumulation of soybean genomic resources, convenient or promising approaches will be requisite for the determination and use of gene function in soybean. In this article, we describe recent advances in and problems of soybean transformation, and survey the current transgenic approaches for applied and basic research in Japan. PMID:23136488

  16. Single Step Stone-Wales Transformation Linking Two Thermodynamically Stable Sc2O@C78 Isomers.

    PubMed

    Zhao, Pei; Li, Meng-Yang; Guo, Yi-Jun; Zhao, Rui-Sheng; Zhao, Xiang

    2016-03-01

    Among the very recently reported dimetallic oxide fullerenes Sc2O@C2n (n = 35-47), a representative Sc2O@C78 still lacks of further characterizations. Herein, a systematical investigation on Sc2O@C78 has been performed by density functional theory combined with statistical thermodynamic studies. Two isolated pentagon rule (IPR) satisfying isomers, Sc2O@D3h(24109)-C78 and Sc2O@C2v(24107)-C78, are disclosed to possess prominent thermodynamic stabilities at the temperature region of fullerene formation. Significantly, these two structures are related by a single Stone-Wales transformation. Moreover, bonding critical points, bond orders, and delocalization indices have been analyzed to uncover covalent interactions in both isomers. In addition, (13)C NMR spectra and UV-vis-NIR adsorptions of the two stable structures are introduced to assist experimental identification and characterization in the future. PMID:26909549

  17. Analysis on DOC transformation in a forested catchment using stable carbon isotope values

    NASA Astrophysics Data System (ADS)

    Ohte, N.; Takahashi, Y.; Itoh, M.; Katsuyama, M.; Fujimoto, M.; Matsuo, N.; Tani, M.

    2009-12-01

    In order to elucidate the production, consumption and transformation mechanisms of dissolved organic carbon (DOC) in temperate forested catchment , time and spatial variations in several parameters which characterize the DOC quality were surveyed in various hydrological processes in a headwater catchment in central Japan. Rain, through fall, soil waters in various depths, groundwater and streamwater were sampled since June 2006 to November 2008. Concentration, δ13C, fluorescence spectrum and UV absorbance of DOC were measured. We measured the δ13C-DOC values using an IRMS with a CO2 purifying system connected to a TOC analyzer. Combined discussions on the profiles of concentration, δ13C and fluorescence characteristics provided following findings: 1) Microbial decomposition of DOC was commonly predominant for consumption in relatively shallower soil horizons (0 to 30 cm in depth) regardless of the soil moisture condition among the soil profiles at the different parts in hillslope, while adsorption was significant in the relatively dry soil profile. 2) During this process, persistent portions remained preferentially in the soil solution. 3) In the groundwater body, two different processes caused; protein-like dissolved organic carbon was added in the relatively oxygen rich part and the δ13C value decreased with the anoxic DOC decomposition in the oxygen poor part, suggesting the methanogenic activity related DOC production (Figure 1). Stream DOC characteristics were determined by relative contributions of subsurface water and groundwater. Then, it received fresh DOC with high fulvic fluorescence peak from bank side or riparian zones again. These indicated that the stream DOC characteristics were influenced not only by in stream and streamside organic supply, but also terrestrial DOC formation through the groundwater discharge. Figure 1. Relationship between UV/DOC and δ13C-DOC of soil water at three different soil profiles. G1, G34 and G27 were located in the

  18. Effects of plant biomass on nitrogen transformation in subsurface-batch constructed wetlands: a stable isotope and mass balance assessment.

    PubMed

    Chen, Yi; Wen, Yue; Zhou, Qi; Vymazal, Jan

    2014-10-15

    Nitrate is commonly found in the influent of subsurface-batch constructed wetlands (SSB CWs) used for tertiary wastewater treatments. To understand the effects of plants and the litter on nitrate removal, as well as on nitrogen transformation in SSB CWs, six laboratory-scale SSB CW microcosms were set up in duplicate and were operated as batch systems with hydraulic residence time (HRT) of 5d. The presence of Typha latifolia enhanced nitrate removal in SSB CWs, and the N removed by plant uptake was mainly stored in aboveground biomass. Typha litter addition greatly improved nitrate removal in SSB CWs through continuous input of labile organic carbon, and calculated enrichment factors (ε) were between -12.1‰--13.9‰ from the nitrogen stable isotope analysis, suggesting that denitrification plays a dominant role in the N removal. Most significantly, simultaneous sulfur-based autotrophic and heterotrophic denitrification was observed in CWs. Finally, mass balance showed that denitrification, sedimentation burial and plant uptake respectively contributed 54%-94%, 1%-46% and 7.5%-14.3% to the N removal in CWs. PMID:25000198

  19. Efficient Agrobacterium tumefaciens-mediated transformation and regeneration of garlic (Allium sativum) immature leaf tissue.

    PubMed

    Kenel, Fernand; Eady, Colin; Brinch, Sheree

    2010-03-01

    Transgenic garlic (Allium sativum) plants have been recovered directly from immature leaf material by selective culture following Agrobacterium-mediated transformation. This method involved the use of a binary vector containing the mgfp-ER reporter gene and hpt selectable marker, and followed a similar protocol developed previously for the transformation of immature onion embryos. The choice of tissue and post-transformation selection procedure resulted in a large increase in recovery of transgenic plants compared with previously confirmed allium transformation protocols. The presence of transgenes in the genome of the plants was confirmed using Southern analysis. This improvement in frequency and the use of clonal commercial "Printanor" germplasm now makes possible the integration of useful agronomic and quality traits into this crop. PMID:20099065

  20. In planta transformation method for T-DNA transfer in orchids

    NASA Astrophysics Data System (ADS)

    Semiarti, Endang; Purwantoro, Aziz; Mercuriani, Ixora S.; Anggriasari, Anida M.; Jang, Seonghoe; Suhandono, Sony; Machida, Yasunori; Machida, Chiyoko

    2014-03-01

    Transgenic plant technology is an efficient tool to study the function of gene(s) in plant. The most popular and widely used technique is Agrobacterium-mediated transformation in which cocultivation was done by immersing the plant tissues/organ in overnight bacterial cultured for about 30 minutes to one hour under in vitro condition. In this experiment, we developed more easier technique that omitted the in vitro step during cocultivation with Agrobacterium, namely in planta transformation method. Pollinaria (compact pollen mass of orchid) of Phalaenopsis amabilis and Spathoglottis plicata orchids were used as target explants that were immersed into bacterial culture for 30 minutes, then dried up the pollinaria, the transformed pollinaria was used to pollinate orchid flowers. The T-DNA used for this experiments were Ubipro∷PaFT/A. tumefaciens GV3101 for P. amabilis and MeEF1α2 pro∷GUS/ A. tumefaciens LBA 4404 for S.plicata. Seeds that were produced from pollinated flowers were grown onto 10 mg/l hygromicin containing NP (New Phalaenopsis) medium. The existance of transgene in putative transformant protocorm (developing orchid embryo) genome was confirmed using PCR with specific primers of either PaFT or GUS genes. Histochemical GUS assay was also performed to the putative transformants. The result showed that transformation frequencies were 2.1 % in P. amabilis, and 0,53% in S. plicata. These results indicates that in planta transformation method could be used for Agrobacterium-mediated genetic transformation, with advantage easier and more secure work from contaminants than that of the in vitro method.

  1. TRANSFORMATION

    SciTech Connect

    LACKS,S.A.

    2003-10-09

    Transformation, which alters the genetic makeup of an individual, is a concept that intrigues the human imagination. In Streptococcus pneumoniae such transformation was first demonstrated. Perhaps our fascination with genetics derived from our ancestors observing their own progeny, with its retention and assortment of parental traits, but such interest must have been accelerated after the dawn of agriculture. It was in pea plants that Gregor Mendel in the late 1800s examined inherited traits and found them to be determined by physical elements, or genes, passed from parents to progeny. In our day, the material basis of these genetic determinants was revealed to be DNA by the lowly bacteria, in particular, the pneumococcus. For this species, transformation by free DNA is a sexual process that enables cells to sport new combinations of genes and traits. Genetic transformation of the type found in S. pneumoniae occurs naturally in many species of bacteria (70), but, initially only a few other transformable species were found, namely, Haemophilus influenzae, Neisseria meningitides, Neisseria gonorrheae, and Bacillus subtilis (96). Natural transformation, which requires a set of genes evolved for the purpose, contrasts with artificial transformation, which is accomplished by shocking cells either electrically, as in electroporation, or by ionic and temperature shifts. Although such artificial treatments can introduce very small amounts of DNA into virtually any type of cell, the amounts introduced by natural transformation are a million-fold greater, and S. pneumoniae can take up as much as 10% of its cellular DNA content (40).

  2. TRANSFORMER

    DOEpatents

    Baker, W.R.

    1959-08-25

    Transformers of a type adapted for use with extreme high power vacuum tubes where current requirements may be of the order of 2,000 to 200,000 amperes are described. The transformer casing has the form of a re-entrant section being extended through an opening in one end of the cylinder to form a coaxial terminal arrangement. A toroidal multi-turn primary winding is disposed within the casing in coaxial relationship therein. In a second embodiment, means are provided for forming the casing as a multi-turn secondary. The transformer is characterized by minimized resistance heating, minimized external magnetic flux, and an economical construction.

  3. Reliable transformation system for Microbotryum lychnidis-dioicae informed by genome and transcriptome project.

    PubMed

    Toh, Su San; Treves, David S; Barati, Michelle T; Perlin, Michael H

    2016-10-01

    Microbotryum lychnidis-dioicae is a member of a species complex infecting host plants in the Caryophyllaceae. It is used as a model system in many areas of research, but attempts to make this organism tractable for reverse genetic approaches have not been fruitful. Here, we exploited the recently obtained genome sequence and transcriptome analysis to inform our design of constructs for use in Agrobacterium-mediated transformation techniques currently available for other fungi. Reproducible transformation was demonstrated at the genomic, transcriptional and functional levels. Moreover, these initial proof-of-principle experiments provide evidence that supports the findings from initial global transcriptome analysis regarding expression from the respective promoters under different growth conditions of the fungus. The technique thus provides for the first time the ability to stably introduce transgenes and over-express target M. lychnidis-dioicae genes. PMID:27215216

  4. Comparison of Soybean Transformation Efficiency and Plant Factors Affecting Transformation during the Agrobacterium Infection Process

    PubMed Central

    Jia, Yuying; Yao, Xingdong; Zhao, Mingzhe; Zhao, Qiang; Du, Yanli; Yu, Cuimei; Xie, Futi

    2015-01-01

    The susceptibility of soybean genotype to Agrobacterium infection is a key factor for the high level of genetic transformation efficiency. The objective of this study is to evaluate the plant factors related to transformation in cotyledonary nodes during the Agrobacterium infection process. This study selected three genotypes (Williams 82, Shennong 9 and Bert) with high transformation efficiency, which presented better susceptibility to Agrobacterium infection, and three low transformation efficiency genotypes (General, Liaodou 16 and Kottman), which showed a relatively weak susceptibility. Gibberellin (GA) levels and soybean GA20ox2 and CYP707A2 transcripts of high-efficiency genotypes increased and were higher than those of low-efficiency genotypes; however, the opposite performance was shown in abscisic acid (ABA). Higher zeatin riboside (ZR) content and DNA quantity, and relatively higher expression of soybean IPT5, CYCD3 and CYCA3 were obtained in high-efficiency genotypes. High-efficiency genotypes had low methyl jasmonate (MeJA) content, polyphenol oxidase (PPO) and peroxidase (POD) activity, and relatively lower expression of soybean OPR3, PPO1 and PRX71. GA and ZR were positive plant factors for Agrobacterium-mediated soybean transformation by facilitating germination and growth, and increasing the number of cells in DNA synthesis cycle, respectively; MeJA, PPO, POD and ABA were negative plant factors by inducing defence reactions and repressing germination and growth, respectively. PMID:26262617

  5. Stable transformation of the gram-positive phytopathogenic bacterium Clavibacter michiganensis subsp. sepedonicus with several cloning vectors.

    PubMed Central

    Laine, M J; Nakhei, H; Dreier, J; Lehtilä, K; Meletzus, D; Eichenlaub, R; Metzler, M C

    1996-01-01

    In this paper we describe transformation of Clavibacter michiganensis subsp. sepedonicus, the potato ring rot bacterium, with plasmid vectors. Three of the plasmids used, pDM100, pDM302, and pDM306, contain the origin of replication from pCM1, a native plasmid of C. michiganensis subsp. michiganensis. We constructed two new cloning vectors, pHN205 and pHN216, by using the origin of replication of pCM2, another native plasmid of C. michiganensis subsp. michiganensis. Plasmids pDM302, pHN205, and pHN216 were stably maintained without antibiotic selection in various strains of C. michiganensis subsp. sepedonicus. We observed that for a single plasmid, different strains of C. michiganensis subsp. sepedonicus showed significantly different transformation efficiencies. We also found unexplained strain-to-strain differences in stability with various plasmid constructions containing different arrangements of antibiotic resistance genes and origins of replication. We examined the effect of a number of factors on transformation efficiency. The best transformation efficiencies were obtained when C. michiganensis subsp. sepedonicus cells were grown on DM agar plates, harvested during the early exponential growth phase, and used fresh (without freezing) for electroporation. The maximal transformation efficiency obtained was 4.6 x 10(4) CFU/microgram of pHN216 plasmid DNA. To demonstrate the utility of this transformation system, we cloned a beta-1,4-endoglucanase-encoding gene from C. michiganensis subsp. sepedonicus into pHN216. When this construction, pHN216:C8, was electroporated into competent cells of a cellulase-deficient mutant, it restored cellulase production to almost wild-type levels. PMID:8633849

  6. Maize transformation technology development for commercial event generation

    PubMed Central

    Que, Qiudeng; Elumalai, Sivamani; Li, Xianggan; Zhong, Heng; Nalapalli, Samson; Schweiner, Michael; Fei, Xiaoyin; Nuccio, Michael; Kelliher, Timothy; Gu, Weining; Chen, Zhongying; Chilton, Mary-Dell M.

    2014-01-01

    Maize is an important food and feed crop in many countries. It is also one of the most important target crops for the application of biotechnology. Currently, there are more biotech traits available on the market in maize than in any other crop. Generation of transgenic events is a crucial step in the development of biotech traits. For commercial applications, a high throughput transformation system producing a large number of high quality events in an elite genetic background is highly desirable. There has been tremendous progress in Agrobacterium-mediated maize transformation since the publication of the Ishida et al. (1996) paper and the technology has been widely adopted for transgenic event production by many labs around the world. We will review general efforts in establishing efficient maize transformation technologies useful for transgenic event production in trait research and development. The review will also discuss transformation systems used for generating commercial maize trait events currently on the market. As the number of traits is increasing steadily and two or more modes of action are used to control key pests, new tools are needed to efficiently transform vectors containing multiple trait genes. We will review general guidelines for assembling binary vectors for commercial transformation. Approaches to increase transformation efficiency and gene expression of large gene stack vectors will be discussed. Finally, recent studies of targeted genome modification and transgene insertion using different site-directed nuclease technologies will be reviewed. PMID:25140170

  7. Stable and Metastable InGaAs/GaAs Island Shapes and Surfactant-like Suppression of the Wetting Transformation

    NASA Technical Reports Server (NTRS)

    Leon, R.; Lobo, C.; Zou, J.; Romeo, T.; Cockayne, D. J. H.

    1998-01-01

    Diverging behaviors are observed in the InGaAs/GaAs Stranski-Krastanow (S-K) island formation during vapor phase epitaxy: varying group V partial pressures gives different critical thicknesses for the onset of the S-K transformation, island surface coverages, ratios between coherent and incoherent islands, and dissimilar morphologies upon annealing.

  8. Coupled transformation of inorganic stable carbon-13 and nitrogen-15 isotopes into higher trophic levels in a eutrophic shallow lake

    SciTech Connect

    Not Available

    1985-07-01

    Enclosure and bag experiments were done in a eutrophic shallow lake with simultaneous use of inorganic /sup 13/C and /sup 15/N isotopes. It was demonstrated that coupled transformation of inorganic carbon and nitrogen can occur into herbivorous zooplankton through phytoplankton. Direct evidence is provided that there is an apparent coupling between photosynthesis and organic nitrogen uptake by phytoplankton during daytime under natural conditions and that the coupling occurs at a constant ratio.

  9. [Genetic transformation of flax (Linum usitatissimum L.) with chimeric GFP-TUA6 gene for visualisation of microtubules].

    PubMed

    Shisha, E N; Korkhovoĭ, V I; Baer, G Ia; Guzenko, E V; Lemesh, V A; Kartel', N A; Emets, A I; Blium, Ia B

    2013-01-01

    The data of Agrobacterium-mediated transformation of some Linum usitatissimum cultivars zoned on the territories of Belarus and Ukraine with the plasmid carrying chimeric GFP-TUA6 gene and nptII gene as selectable marker conferring resistance to kanamycin are presented in this study. Transformation was affected by a number of factors including optical density (OD600), time of inoculation of explants with Agrobacterium and co-culture conditions. Transgenic nature of obtained lines was confirmed by PCR analysis. Expression of GFP-TUA6 gene was detected with confocal laser scanning microscopy. The obtained transgenic lines can be used for further functional studies the role of microtubules in the processes of building the flax fibres and resistance to wind. PMID:23745358

  10. Sources, migration and transformation of antimony contamination in the water environment of Xikuangshan, China: Evidence from geochemical and stable isotope (S, Sr) signatures.

    PubMed

    Wen, Bing; Zhou, Jianwei; Zhou, Aiguo; Liu, Cunfu; Xie, Lina

    2016-11-01

    The Xikuangshan (XKS) mine in central China is the largest antimony (Sb) mine in the world. The mining activity has seriously contaminated the waters in the area. To determine the sources, migration and transformation of Sb contamination, 32 samples from groundwater (aquifer water), surface water and mine water were collected for water chemistry, trace element and SSO4 and Sr stable isotope analyses. The results showed that the groundwater and surface water were in an oxidized environment. The SSO4 and Sr isotope compositions in the water indicated that dissolved Sb and SO4(2) originated from sulfide mineral (Sb2S3) oxidation, whereas radiogenic Sr may have been sourced from silicified limestone and stibnite in the Shetianqiao aquifer. Furthermore, a positive correlation between δ(34)SSO4 and δ(87)Sr values revealed that the Sr, S and Sb in the waters had a common contamination source, i.e., silicified limestone and stibnite, whereas the Sr, S and Sb in rock and ore were sourced from Proterozoic basement clastics. The analysis also indicated that the isotope composition of dissolved SO4(2-) had been influenced by slight bacterial SO4 reduction in the Magunao aquifer. Mining or rock collapse may have caused Shetianqiao aquifer water to contaminate the Magunao aquifer water via mixing. This study has demonstrated that the stable isotopes of (34)SSO4 and (87)Sr, combined with hydrochemical methods, are effective in tracking the sources, migration and transformation of Sb contamination. PMID:27341112

  11. Multi-frequency weak signal detection based on wavelet transform and parameter compensation band-pass multi-stable stochastic resonance

    NASA Astrophysics Data System (ADS)

    Han, Dongying; li, Pei; An, Shujun; Shi, Peiming

    2016-03-01

    In actual fault diagnosis, useful information is often submerged in heavy noise, and the feature information is difficult to extract. A novel weak signal detection method aimed at the problem of detecting multi-frequency signals buried under heavy background noise is proposed based on wavelet transform and parameter compensation band-pass multi-stable stochastic resonance (SR). First, the noisy signal is processed by parameter compensation, with the noise and system parameters expanded 10 times to counteract the effect of the damping term. The processed signal is decomposed into multiple signals of different scale frequencies by wavelet transform. Following this, we adjust the size of the scaled signals' amplitudes and reconstruct the signals; the weak signal frequency components are then enhanced by multi-stable stochastic resonance. The enhanced components of the signal are processed through a band-pass filter, leaving the enhanced sections of the signal. The processed signal is analyzed by FFT to achieve detection of the multi-frequency weak signals. The simulation and experimental results show that the proposed method can enhance the signal amplitude, can effectively detect multi-frequency weak signals buried under heavy noise and is valuable and usable for bearing fault signal analysis.

  12. Stable isotope fractionation in photosynthesis: Analysis of autotrophic competence following transformation of the chloroplast genome of Chlamydomonas

    SciTech Connect

    Boynton, J.E.; Gillham, N.W.; Osmond, C.B.

    1991-06-15

    Isotopic techniques needed to assess the interactions between photosynthesis and respiration in Chlamydomonas have been devised for {sup 13}C, using plate and liquid cultures. The effectiveness of various transformation strategies for the chloroplast psbA gene has been evaluated with respect to their utility in constructing and characterizing strains homoplasmic for site-directed mutations in an otherwise isogenic background. Our analysis of the first site-directed change in the D-1 protein of Chlamydomonas indicates that a second site mutation (arg{sub 238} > lys) in the loop between transmembrane helices IV -- V can partially compensate for the reduced photosynthetic performance that accompanies the atrazine resistant mutation (ser{sub 264} > ala/gly) in this alga and in higher plants grown under high light intensities. 31 refs., 2 figs.

  13. Creation and validation of a widely applicable multiple gene transfer vector system for stable transformation in plant.

    PubMed

    Sun, Quanxi; Liu, Jiang; Li, Yaxiao; Zhang, Qin; Shan, Shihua; Li, Xinzheng; Qi, Baoxiu

    2013-11-01

    Multiple gene transfer (MGT) technology has become a powerful tool for basic and applied plant biology research in recent years. Despite some notable successes in obtaining plant lines harbouring multiple transgenes, these methods are still generally unwieldy and costly. We report here a straightforward and cost effective strategy, utilizing commonly available restriction enzymes for the transfer of multiple genes into plants, hence greatly widening the accessibility of MGT. This methodology exploits the specific 'nested' arrangement of a pair of isocaudomer restriction enzymes (for example XbaI-AvrII-XbaI) so that through the alternate use of these two enzymes in a reiterative fashion multiple genes/constructs (up to five in this study) could be 'stacked' together with ease. In a proof-of-concept experiment, we constructed a plant transformation vector containing three reporter gene expression cassettes flanked by two matrix attachment region sequences. The expression of all three genes was confirmed in transgenic Arabidopsis thaliana. The usefulness of this technology was further validated by the construction of a plant transformation vector containing five transgenes for the production of eicosapentaenoic acid (EPA, C20∆⁵,⁸,¹¹,¹⁴,¹⁷), a polyunsaturated essential fatty acid found in fish oils that is beneficial for health. In addition, we constructed four more vectors, incorporating one seed specific and three promoters conferring constitutive expression. These expression cassettes are flanked by a different isocaudomer pair (AvrII-SpeI-AvrII) and four other unique restriction sites, allowing the exchange of promoters and terminators of choice. PMID:23839253

  14. Stable and near Fourier-transform-limit 30fs pulse compression with a tiled grating compressor scheme.

    PubMed

    Li, Zhaoyang; Li, Shuai; Wang, Cheng; Xu, Yi; Wu, Fengxiang; Li, Yanyan; Leng, Yuxin

    2015-12-28

    The fabrication of meter-sized-gratings limits the rapid development of worldwide femtosecond ultra-intense lasers with 10PW or higher peak power, and the grating tiling method is an alternative to this problem. But the unreliability and complexity of the traditional grating tiling, which has already been widely used in picosecond Petawatt lasers, challenges its application in femtosecond ultra-intense lasers for wider bandwidth and more sensitive grating attitude. In this paper, we demonstrate an experiment to compress an 800nm centered nanosecond deep-chirped-pulse to around 30fs by an object-image-grating self-tiling compressor, to the best of our knowledge, which is the shortest pulse compressed by a tiled grating compressor. Both the compression pulse and the focal spot are stable and close to the theoretical limits. We believe it is a feasible solution to the grating-size-limited problem for recent femtosecond 10PW Ti:sapphire or OPCPA laser projects. PMID:26832003

  15. Transformation of a fragment of beta-structural bacteriophage T4 adhesin to stable alpha-helical trimer.

    PubMed

    Miroshnikov, K A; Sernova, N V; Shneider, M M; Mesyanzhinov, V V

    2000-12-01

    Gene product 12 of bacteriophage T4, adhesin, serves to adhere the virus to host cells. Adhesin is a fibrous homotrimer, and a novel tertiary structure element, a beta-helix, is supposed to be a major structural feature of this protein. We have constructed two truncated gp12 mutants, 12N1 and 12N2, containing 221 and 135 N-terminal residues, respectively. When expressed in E. coli cells, these gp12 fragments formed labile beta-structural trimers. Another hybrid protein, 12FN, containing 179 N-terminal amino acid residues of gp12 fused to the C-terminal domain (31 amino acids) of T4 fibritin, was shown to have a trimeric proteolytically resistant alpha-helical structure. This structure is probably similar to that of fibritin, which has a triple alpha-helical coiled-coil structure. Hence, we have demonstrated the possibility of global transformation of fibrous protein structure using fusion with a C-terminal domain that initiates trimerization. PMID:11173503

  16. Real-time field measurements of stable isotopes in water and CO2 by Fourier transform infrared spectrometry.

    PubMed

    Griffith, David W T; Jamie, Ian; Esler, Michael; Wilson, Stephen R; Parkes, Stephen D; Waring, Christopher; Bryant, Glenn W

    2006-03-01

    Continuous records of isotope behaviour in the environment are invaluable to understanding mass and energy fluxes. Although techniques such as isotope ratio mass spectrometry provide high precision data, they are not well suited to the analysis of a large number of samples and are currently restricted to use in the laboratory. Fourier transform infrared spectrometers are relatively cheap and sufficiently portable and robust to be taken into the field to collect continuous records of gas-phase isotope behaviour. Several examples of the application of this technique will be presented. One data set provides half-hourly determinations of vertical profiles of D/H in water vapour above agricultural fields over a 3-week period; the same infrared spectra can also be used to determine 13C/12C in CO2. The technique has also been applied to the study of CO2 in ambient air and in a limestone cave system. Some of the features and complications associated with the method will also be considered. PMID:16500751

  17. Transformation of Silicon in a Coastal Sandy Beach Ecosystem: Insights from Stable Silicon Isotopes in Ground- and Porewaters

    NASA Astrophysics Data System (ADS)

    Ehlert, C.; Paffrath, R.; Reckhardt, A.; Pahnke, K.

    2014-12-01

    Wave and tide dominated sandy beach ecosystems have a major effect on coastal carbon, nutrient, and trace element cycling. The coarse-grained sediments are characterized by a high permeability, which permits advective porewater transport. Due to the fast porewater flow in the sandy sediment, organic matter remineralization products (i.e. nutrients and trace metals) are only slightly enriched in the porewater and rapidly discharged to the coastal zone where they can potentially stimulate primary production. Here we present stable silicon isotope (δ30Si) data from groundwaters and porewaters from Spiekeroog Island, a barrier island in the southern North Sea, Germany. Groundwater was sampled in the freshwater lens down to 70 m depth in the center of the island. The beach porewaters were taken along a transect from the dune belt to the low water line at depths between 0.5 and 5.2 m. This transect represents the mixing zone of fresh and saline groundwater with seawater, whereby the terrestrial driven groundwater flow provides a potential source of terrestrially derived Si isotope compositions resulting in a modification of the marine-dominated porewater composition. The groundwater is characterized by high Si concentrations of up to 750 µM and rather low δ30Si signatures between +0.6‰ and +1.2‰, which decrease with depth (and age) within the freshwater lens. The porewaters have much lower Si concentrations around 50-60 µM and higher and more variable δ30Si between +1.0‰ and +2.5‰. The lower δ30Si values are found closer to the dunes and at greater depths in the beach sediments. These signatures possibly reflect the outflow of the fresh groundwater to the coastal zone. The higher values represent the influence of seawater, which delivers water with a high δ30Si signature previously enriched by phytoplankton growth.

  18. Aquatic photochemistry, abiotic and aerobic biodegradability of thalidomide: identification of stable transformation products by LC-UV-MS(n).

    PubMed

    Mahmoud, Waleed M M; Trautwein, Christoph; Leder, Christoph; Kümmerer, Klaus

    2013-10-01

    Thalidomide (TD), besides being notorious for its teratogenicity, was shown to have immunomodulating and anti-inflammatory activities. This is why recently TD became a promising drug for the treatment of different cancers and inflammatory diseases. Yet nothing is known about the environmental fate of TD, which therefore was assessed experimentally and by in silico prediction programs (quantitative structure activity relationship (QSAR) models) within this study. Photolytic degradation was tested with two different light sources (medium-pressure mercury lamp; xenon lamp) and aerobic biodegradability was investigated with two OECD tests (Closed Bottle test (CBT), Manometric Respirometry test (MRT)). An additional CBT was performed for TD samples after 16 min of UV-photolysis. The primary elimination of TD was monitored and the structures of its photo-, abiotic and biodegradation products were elucidated by HPLC-UV-Fluorescence-MS(n). Furthermore, elimination of dissolved organic carbon was monitored in the photolysis experiment. LC-MS revealed that new photolytic transformation products (TPs) were identified, among them two isomers of TD with the same molecular mass. These TPs were different to the products formed by biodegradation. The experimental findings were compared with the results obtained from the in silico prediction programs where e.g. a good correlation for TD biodegradation in the CBT was confirmed. Moreover, some of the identified TPs were also structurally predicted by the MetaPC software. These results demonstrate that TD and its TPs are not readily biodegradable and not fully mineralized by photochemical treatment. They may therefore pose a risk to the aquatic environment due to the pharmacological activity of TD and unknown properties of its TPs. The applied techniques within this study emphasize the importance of QSAR models as a tool for estimating environmental risk assessments. PMID:23792256

  19. Pathways and transformations of dissolved methane and dissolved inorganic carbon in Arctic tundra soils: Evidence from analysis of stable isotopes

    NASA Astrophysics Data System (ADS)

    Throckmorton, H.; Perkins, G.; Muss, J. D.; Smith, L. J.; Conrad, M. E.; Torn, M. S.; Heikoop, J. M.; Newman, B. D.; Wilson, C. J.; Wullschleger, S. D.

    2014-12-01

    Arctic soils contain a large pool of terrestrial C and are of great interest because of their potential for releasing significant amounts of carbon dioxide (CO2) and methane (CH4) to the atmosphere. Few attempts have been made, however, to derive quantitative budgets of CO2 and CH4 budgets for high-latitude ecosystems. Therefore, this study used naturally occurring geochemical and isotopic tracers to estimate production pathways and transformations of dissolved inorganic carbon (DIC = Σ (total) dissolved CO2) and dissolved CH4 in soil pore waters from 17 locations (drainages) in Barrow, Alaska (USA) in July and September, 2013; and to approximate a complete balance of belowground C cycling at our sampling locations. Results suggest that CH4 was primarily derived from biogenic acetate fermentation, with a shift at 4 locations from July to September towards CO2 reduction as the dominant methanogenic pathway. A large majority of CH4 produced at the frost table methane was transferred directly to the atmosphere via plant roots and ebullition (94.0 ± 1.4% and 96.6 ± 5.0% in July and September). A considerable fraction of the remaining CH4 was oxidized to CO2 during upward diffusion in July and September, respectively. Methane oxidization produced <1% of CO2 relative to alternative production mechanisms in deep subsurface pore waters. The majority of subsurface CO2 was produced from anaerobic respiration, likely due to reduction of Fe oxides and humics (52 ± 6 to 100 ± 13%, on average) while CO2 produced from methanogenesis accounted for the remainder (0 ± 13% to 47 ± 6%, on average) for July and September, respectively. Dissolved CH4 and dissolved CO2 concentrations correlated with thaw depth, suggesting that Arctic ecosystems will likely produce and release a greater amount of greenhouse gasses under projected warming and deepening of active layer thaw depth under future climate change scenarios.

  20. A new approach to understand methylmercury (CH3Hg) sources and transformation pathways: Compound-specific carbon stable isotope analysis by GC-C-IRMS

    NASA Astrophysics Data System (ADS)

    Baya, P. A.; Point, D.; Amouroux, D. P.; Lebreton, B.; Guillou, G.

    2015-12-01

    Methylmercury (CH3Hg) is a potent neurotoxin which is readily assimilated by organisms and bio-accumulates in aquatic food webs. In humans, consumption of CH3Hg contaminated marine fish is the major route of mercury exposure. However, our understanding of CH3Hg transformation pathways is still incomplete. To close this knowledge gap, we propose to explore the stable carbon isotopic composition (δ13C) of the methyl group of CH3Hg for a better understanding of its sources and transformation mechanisms. The method developed for the determination of the δ13C value of CH3Hg in biological samples involves (i) CH3Hg selective extraction, (ii) derivatization, and (iii) separation by gas chromatography (GC) prior to analysis by combustion isotope ratio mass spectrometry (C-IRMS). We present the figures of merit of this novel method and the first δ13C signatures for certified materials (ERM-CE464, BCR414) and biological samples at different marine trophic levels (i.e., tuna fish, zooplankton). The implications of this new approach to trace the pathways associated with Hg methylation and the mechanisms involved will be discussed.

  1. Stable transformation and reverse genetic analysis of Penium margaritaceum: a platform for studies of charophyte green algae, the immediate ancestors of land plants.

    PubMed

    Sørensen, Iben; Fei, Zhangjun; Andreas, Amanda; Willats, William G T; Domozych, David S; Rose, Jocelyn K C

    2014-02-01

    The charophyte green algae (CGA, Streptophyta, Viridiplantae) occupy a key phylogenetic position as the immediate ancestors of land plants but, paradoxically, are less well-studied than the other major plant lineages. This is particularly true in the context of functional genomic studies, where the lack of an efficient protocol for their stable genetic transformation has been a major obstacle. Observations of extant CGA species suggest the existence of some of the evolutionary adaptations that had to occur for land colonization; however, to date, there has been no robust experimental platform to address this genetically. We present a protocol for high-throughput Agrobacterium tumefaciens-mediated transformation of Penium margaritaceum, a unicellular CGA species. The versatility of Penium as a model for studying various aspects of plant cell biology and development was illustrated through non-invasive visualization of protein localization and dynamics in living cells. In addition, the utility of RNA interference (RNAi) for reverse genetic studies was demonstrated by targeting genes associated with cell wall modification (pectin methylesterase) and biosynthesis (cellulose synthase). This provided evidence supporting current models of cell wall assembly and inter-polymer interactions that were based on studies of land plants, but in this case using direct observation in vivo. This new functional genomics platform has broad potential applications, including studies of plant organismal biology and the evolutionary innovations required for transition from aquatic to terrestrial habitats. PMID:24308430

  2. Floral-dip transformation of flax (Linum usitatissimum) to generate transgenic progenies with a high transformation rate.

    PubMed

    Bastaki, Nasmah K; Cullis, Christopher A

    2014-01-01

    Agrobacterium-mediated plant transformation via floral-dip is a widely used technique in the field of plant transformation and has been reported to be successful for many plant species. However, flax (Linum usitatissimum) transformation by floral-dip has not been reported. The goal of this protocol is to establish that Agrobacterium and the floral-dip method can be used to generate transgenic flax. We show that this technique is simple, inexpensive, efficient, and more importantly, gives a higher transformation rate than the current available methods of flax transformation. In summary, inflorescences of flax were dipped in a solution of Agrobacterium carrying a binary vector plasmid (T-DNA fragment plus the Linum Insertion Sequence, LIS-1) for 1 - 2 min. The plants were laid flat on their side for 24 hr. Then, plants were maintained under normal growth conditions until the next treatment. The process of dipping was repeated 2 - 3 times, with approximately 10 - 14 day intervals between dipping. The T1 seeds were collected and germinated on soil. After approximately two weeks, treated progenies were tested by direct PCR; 2 - 3 leaves were used per plant plus the appropriate T-DNA primers. Positive transformants were selected and grown to maturity. The transformation rate was unexpectedly high, with 50 - 60% of the seeds from treated plants being positive transformants. This is a higher transformation rate than those reported for Arabidopsis thaliana and other plant species, using floral-dip transformation. It is also the highest, which has been reported so far, for flax transformation using other methods for transformation. PMID:25549243

  3. How Stable Is Stable?

    ERIC Educational Resources Information Center

    Baehr, Marie

    1994-01-01

    Provides a problem where students are asked to find the point at which a soda can floating in some liquid changes its equilibrium between stable and unstable as the soda is removed from the can. Requires use of Newton's first law, center of mass, Archimedes' principle, stable and unstable equilibrium, and buoyant force position. (MVL)

  4. Efficient sweet pepper transformation mediated by the BABY BOOM transcription factor.

    PubMed

    Heidmann, Iris; de Lange, Brenda; Lambalk, Joep; Angenent, Gerco C; Boutilier, Kim

    2011-06-01

    Pepper (Capsicum L.) is a nutritionally and economically important crop that is cultivated throughout the world as a vegetable, condiment, and food additive. Genetic transformation using Agrobacterium tumefaciens (agrobacterium) is a powerful biotechnology tool that could be used in pepper to develop community-based functional genomics resources and to introduce important agronomic traits. However, pepper is considered to be highly recalcitrant for agrobacterium-mediated transformation, and current transformation protocols are either inefficient, cumbersome or highly genotype dependent. The main bottleneck in pepper transformation is the inability to generate cells that are competent for both regeneration and transformation. Here, we report that ectopic expression of the Brassica napus BABY BOOM AP2/ERF transcription factor overcomes this bottleneck and can be used to efficiently regenerate transgenic plants from otherwise recalcitrant sweet pepper (C. annuum) varieties. Transient activation of BABY BOOM in the progeny plants induced prolific cell regeneration and was used to produce a large number of somatic embryos that could be converted readily to seedlings. The data highlight the utility of combining biotechnology and classical plant tissue culture approaches to develop an efficient transformation and regeneration system for a highly recalcitrant vegetable crop. PMID:21305301

  5. Efficient Transformation of Oil Palm Protoplasts by PEG-Mediated Transfection and DNA Microinjection

    PubMed Central

    Masani, Mat Yunus Abdul; Noll, Gundula A.; Parveez, Ghulam Kadir Ahmad; Sambanthamurthi, Ravigadevi; Prüfer, Dirk

    2014-01-01

    Background Genetic engineering remains a major challenge in oil palm (Elaeis guineensis) because particle bombardment and Agrobacterium-mediated transformation are laborious and/or inefficient in this species, often producing chimeric plants and escapes. Protoplasts are beneficial as a starting material for genetic engineering because they are totipotent, and chimeras are avoided by regenerating transgenic plants from single cells. Novel approaches for the transformation of oil palm protoplasts could therefore offer a new and efficient strategy for the development of transgenic oil palm plants. Methodology/Principal Findings We recently achieved the regeneration of healthy and fertile oil palms from protoplasts. Therefore, we focused on the development of a reliable PEG-mediated transformation protocol for oil palm protoplasts by establishing and validating optimal heat shock conditions, concentrations of DNA, PEG and magnesium chloride, and the transfection procedure. We also investigated the transformation of oil palm protoplasts by DNA microinjection and successfully regenerated transgenic microcalli expressing green fluorescent protein as a visible marker to determine the efficiency of transformation. Conclusions/Significance We have established the first successful protocols for the transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection. These novel protocols allow the rapid and efficient generation of non-chimeric transgenic callus and represent a significant milestone in the use of protoplasts as a starting material for the development of genetically-engineered oil palm plants. PMID:24821306

  6. THE K-REGION DIHYDRODIOL OF BENZO[A]PYRENE INDUCES DNA DAMAGE AND MORPHOLOGICAL CELL TRANSFORMATION IN C3H10T1/2CL8 MOUSE EMBRYO CELLS WITHOUT THE FORMATION OF DETECTABLE STABLE COVALENT DNA ADDUCTS

    EPA Science Inventory

    The K -region dihydrodiol ofbenzo[ a ]pyrene induces DNA damage and morphological cell transformation in C3HlOTY2CL8 mouse embryo cells without the formation of detectable stable covalent DNA adducts

    Benzo[ a ]pyrene (B[ a ]P) is the most thoroughly studied polycyclic aro...

  7. Progress of cereal transformation technology mediated by Agrobacterium tumefaciens

    PubMed Central

    Hiei, Yukoh; Ishida, Yuji; Komari, Toshihiko

    2014-01-01

    Monocotyledonous plants were believed to be not transformable by the soil bacterium Agrobacterium tumefaciens until two decades ago, although convenient protocols for infection of leaf disks and subsequent regeneration of transgenic plants had been well established in a number of dicotyledonous species by then. This belief was reinforced by the fact that monocotyledons are mostly outside the host range of crown gall disease caused by the bacterium and by the failures in trials in monocotyledons to mimic the transformation protocols for dicotyledons. However, a key reason for the failure could have been the lack of active cell divisions at the wound sites in monocotyledons. The complexity and narrow optimal windows of critical factors, such as genotypes of plants, conditions of the plants from which explants are prepared, tissue culture methods and culture media, pre-treatments of explants, strains of A. tumefaciens, inducers of virulence genes, transformation vectors, selection marker genes and selective agents, kept technical hurdles high. Eventually it was demonstrated that rice and maize could be transformed by co-cultivating cells of callus cultures or immature embryos, which are actively dividing or about to divide, with A. tumefaciens. Subsequently, these initial difficulties were resolved one by one by many research groups, and the major cereals are now transformed quite efficiently. As many as 15 independent transgenic events may be regenerated from a single piece of immature embryo of rice. Maize transformation protocols are well established, and almost all transgenic events deregulated for commercialization after 2003 were generated by Agrobacterium-mediated transformation. Wheat, barley, and sorghum are also among those plants that can be efficiently transformed by A. tumefaciens. PMID:25426132

  8. Drought-tolerant rice germplasm developed from an Oryza officinalis transformation-competent artificial chromosome clone.

    PubMed

    Liu, R; Zhang, H H; Chen, Z X; Shahid, M Q; Fu, X L; Liu, X D

    2015-01-01

    Oryza officinalis has proven to be a natural gene reservoir for the improvement of domesticated rice as it carries many desirable traits; however, the transfer of elite genes to cultivated rice by conventional hybridization has been a challenge for rice breeders. In this study, the conserved sequence of plant stress-related NAC transcription factors was selected as a probe to screen the O. officinalis genomic transformation-competent artificial chromosome library by Southern blot; 11 positive transformation-competent artificial chromosome clones were subsequently detected. By Agrobacterium-mediated transformation, an indica rice variety, Huajingxian 74 (HJX74), was transformed with a TAC clone harboring a NAC gene-positive genomic fragment from O. officinalis. Molecular analysis revealed that the O. officinalis genomic fragment was integrated into the genome of HJX74. The transgenic lines exhibited high tolerance to drought stress. Our results demonstrate that the introduction of stress-related transformation-competent artificial chromosome clones, coupled with a transgenic validation approach, is an effective method of transferring agronomically important genes from O. officinalis to cultivated rice. PMID:26535682

  9. Co-transformation of canola by chimeric chitinase and tlp genes towards improving resistance to Sclerotinia sclerotiorum.

    PubMed

    Aghazadeh, Rustam; Zamani, Mohammadreza; Motallebi, Mostafa; Moradyar, Mehdi; Moghadassi Jahromi, Zahra

    2016-09-01

    Canola (Brassica napus) plants were co-transformed with two pathogenesis-related protein genes expressing a Trichoderma atroviride chitinase with a chitin-binding domain (chimeric chitinase) and a thaumatin-like protein (tlp) from Oryza sativa conferring resistance to phytopatogenic fungi by Agrobacterium-mediated transformation. The putative transgenic plants were confirmed by PCR. After measuring the specific activity of the chimeric chitinase and glucanase activity for tlp genes, transgenic plants with high specific activity were selected for southern blot analysis to confirm the copy number of the genes. In vitro assays, the antifungal activity of crude extracted protein against Sclerotinia sclerotiorum showed that the inhibition percentage in double transgenic plants was between 55 and 62, whereas the inhibition percentage in single-gene transformants (chimeric chitinase) ranged from 35 to 45 percent. Importantly, in greenhouse conditions, the double transgenic plants showed significant resistance than the single-gene transformant and wild type plants. The results in T2 generation using the intact leaf inoculation method showed that the average lesion diameters were 10, 14.7 and 29 mm for the double transformant, single-gene transformant and non-transgenic plants, respectively. Combined expression of chimeric chitinase and tlp in transgenic plants showed significantly enhanced resistance against S. sclerotiorum than the one that express single-gene transformant plants. These results suggest that the co-expression of chimeric chitinase and tlp can confer enhanced disease resistance in canola plant. PMID:27430511

  10. Fibroblast growth factor receptor splice variants are stable markers of oncogenic transforming growth factor β1 signaling in metastatic breast cancers

    PubMed Central

    2014-01-01

    Introduction Epithelial–mesenchymal transition (EMT) and mesenchymal–epithelial transition (MET) facilitate breast cancer (BC) metastasis; however, stable molecular changes that result as a consequence of these processes remain poorly defined. Therefore, with the hope of targeting unique aspects of metastatic tumor outgrowth, we sought to identify molecular markers that could identify tumor cells that had completed the EMT:MET cycle. Methods An in vivo reporter system for epithelial cadherin (E-cad) expression was used to quantify its regulation in metastatic BC cells during primary and metastatic tumor growth. Exogenous addition of transforming growth factor β1 (TGF-β1) was used to induce EMT in an in situ model of BC. Microarray analysis was employed to examine gene expression changes in cells chronically treated with and withdrawn from TGF-β1, thus completing one full EMT:MET cycle. Changes in fibroblast growth factor receptor type 1 (FGFR1) isoform expression were validated using PCR analyses of patient-derived tumor tissues versus matched normal tissues. FGFR1 gene expression was manipulated using short hairpin RNA depletion and cDNA rescue. Preclinical pharmacological inhibition of FGFR kinase was employed using the orally available compound BGJ-398. Results Metastatic BC cells undergo spontaneous downregulation of E-cad during primary tumor growth, and its expression subsequently returns following initiation of metastatic outgrowth. Exogenous exposure to TGF-β1 was sufficient to drive the metastasis of an otherwise in situ model of BC and was similarly associated with a depletion and return of E-cad expression during metastatic progression. BC cells treated and withdrawn from TGF-β stably upregulate a truncated FGFR1-β splice variant that lacks the outermost extracellular immunoglobulin domain. Identification of this FGFR1 splice variant was verified in metastatic human BC cell lines and patient-derived tumor samples. Expression of FGFR1-β was

  11. Towards the development of better crops by genetic transformation using engineered plant chromosomes.

    PubMed

    Dhar, Manoj K; Kaul, Sanjana; Kour, Jasmeet

    2011-05-01

    Plant Biotechnology involves manipulation of genetic material to develop better crops. Keeping in view the challenges being faced by humanity in terms of shortage of food and other resources, we need to continuously upgrade the genomic technologies and fine tune the existing methods. For efficient genetic transformation, Agrobacterium-mediated as well as direct delivery methods have been used successfully. However, these methods suffer from many disadvantages especially in terms of transfer of large genes, gene complexes and gene silencing. To overcome these problems, recently, some efforts have been made to develop genetic transformation systems based on engineered plant chromosomes called minichromosomes or plant artificial chromosomes. Two approaches namely, "top-down" or "bottom-up" have been used for minichromosomes. The former involves engineering of the existing chromosomes within a cell and the latter de novo assembling of chromosomes from the basic constituents. While some success has been achieved using these chromosomes as vectors for genetic transformation in maize, however, more studies are needed to extend this technology to crop plants. The present review attempts to trace the genesis of minichromosomes and discusses their potential of development into plant artificial chromosome vectors. The use of these vectors in genetic transformation will greatly ameliorate the food problem and help to achieve the UN Millennium development goals. PMID:21249368

  12. Use of Agrobacterium rhizogenes strain 18r12v and paromomycin selection for transformation of Brachypodium distachyon and Brachypodium sylvaticum

    DOE PAGESBeta

    Collier, Ray; Bragg, Jennifer; Hernandez, Bryan T.; Vogel, John P.; Thilmony, Roger

    2016-05-24

    In this study, the genetic transformation of monocot grasses is a resource intensive process, the quality and efficiency of which is dependent in part upon the method of DNA introduction, as well as the ability to effectively separate transformed from wildtype tissue. Agrobacterium-mediated transformation of Brachypodium has relied mainly on Agrobacterium tumefaciens strain AGL1. Currently the antibiotic hygromycin B has been the selective agent of choice for robust identification of transgenic calli in Brachypodium distachyon and Brachypodium sylvaticum but few other chemicals have been shown to work as well for selection of transgenic Brachypodium cells in tissue culture. This studymore » demonstrates that Agrobacterium rhizogenes strain 18r12v and paromomycin selection can be successfully used for the efficient generation of transgenic B. distachyon and B. sylvaticurn. Additionally we observed that the transformation rates were similar to or higher than those obtained with A. turnefaciens strain AGL1 and hygromycin selection. The A. rhizogenes strain 18r12v harboring the pARS1 binary vector and paromomycin selection is an effective means of generating transgenic Brachypodium plants. This novel approach will facilitate the transgenic complementation of T-DNA knockout mutants of B. distachyon which were created using hygromycin selection, as well as aid the implementation of more complex genome manipulation strategies which require multiple rounds of transformation.« less

  13. Use of Agrobacterium rhizogenes Strain 18r12v and Paromomycin Selection for Transformation of Brachypodium distachyon and Brachypodium sylvaticum

    PubMed Central

    Collier, Ray; Bragg, Jennifer; Hernandez, Bryan T.; Vogel, John P.; Thilmony, Roger

    2016-01-01

    The genetic transformation of monocot grasses is a resource intensive process, the quality and efficiency of which is dependent in part upon the method of DNA introduction, as well as the ability to effectively separate transformed from wildtype tissue. Agrobacterium-mediated transformation of Brachypodium has relied mainly on Agrobacterium tumefaciens strain AGL1. Currently the antibiotic hygromycin B has been the selective agent of choice for robust identification of transgenic calli in Brachypodium distachyon and Brachypodium sylvaticum but few other chemicals have been shown to work as well for selection of transgenic Brachypodium cells in tissue culture. This study demonstrates that Agrobacterium rhizogenes strain 18r12v and paromomycin selection can be successfully used for the efficient generation of transgenic B. distachyon and B. sylvaticum. Additionally we observed that the transformation rates were similar to or higher than those obtained with A. tumefaciens strain AGL1 and hygromycin selection. The A. rhizogenes strain 18r12v harboring the pARS1 binary vector and paromomycin selection is an effective means of generating transgenic Brachypodium plants. This novel approach will facilitate the transgenic complementation of T-DNA knockout mutants of B. distachyon which were created using hygromycin selection, as well as aid the implementation of more complex genome manipulation strategies which require multiple rounds of transformation. PMID:27252729

  14. Regulation of astaxanthin and its intermediates through cloning and genetic transformation of β-carotene ketolase in Haematococcus pluvialis.

    PubMed

    Kathiresan, S; Chandrashekar, Arun; Ravishankar, G A; Sarada, R

    2015-02-20

    Astaxanthin, a high-value ketocarotenoid used in the pharmaceutical and nutraceutical industries is mainly produced from green alga, Haematococcus pluvialis. It is biosynthesized by the action of key enzyme, β-carotene ketolase (BKT) on β-carotene through intermediates echinenone and canthaxanthin. In this study, the β-carotene ketolase (bkt) gene was isolated from H. pluvialis and cloned in a vector pRT100 and further mobilized to a binary vector pCAMBIA 1304. The T-DNA of pCAMBIA 1304, which consists of cloned bkt, was successfully transformed to H. pluvialis through Agrobacterium mediation. The cloning and transformation of bkt in H. pluvialis was confirmed by Southern blotting and also by PCR analysis. Total carotenoids and astaxanthin content in the transformed cells were found to be 2-3-fold higher, while the intermediates like echinenone and canthaxanthin were found to be 8-10-fold higher than in the control cells. The expression level of carotenogenic genes like phytoene synthase (psy), phytoene desaturase (pds), lycopene cyclase (lcy), bkt, and β-carotene hydroxylase (bkh) were found to be higher in transformed cells compared to the non-transformed (NT) H. pluvialis. PMID:25612872

  15. Morpho-anatomical characterization of mature embryo-derived callus of rice (Oryza sativa L.) suitable for transformation.

    PubMed

    Bevitori, R; Popielarska-Konieczna, M; dos Santos, E M; Grossi-de-Sá, M F; Petrofeza, S

    2014-05-01

    The objective of this study was to morpho-anatomically characterize embryogenic rice calli during early induction of somatic embryogenesis of three Brazilian rice cultivars. Herein, we explored embryogenic units (EUs) from 2-week-old cut proliferated calli to verify whether they were suitable for Agrobacterium tumefasciens-mediated transformation. Histological analysis and scanning electron microscopy (SEM) were used to analyze these types of calli during early rice callogenesis in the cultivars BRS Primavera, BRS Bonança, and BRS Caiapó. The characteristics of the embryogenic cells were preserved in the EUs, which showed a globular, compact structure that contained tightly packed cells and thus rendered the cells suitable for transformation. The EUs of BRS Caiapó also maintained the characteristics of the non-embryogenic callus, such as an elongated morphology and a lack of cellular organization. In general, the observations of the histological sections corresponded with those of the SEM images. The histological analysis suggested that all cultivars used in these experiments have morphogenic potential. The EUs from proliferated 2-week-old cut calli maintained their embryogenic features. The EUs were subjected to Agrobacterium-mediated transformation, which exhibited a regeneration frequency of 58 % for transformed hygromycin-resistant cell lines. These results show that EUs from proliferated 2-week-old cut calli are suitable for plant transformation. PMID:24085343

  16. Comparison of Fourier Transform Infrared Spectroscopy (FTIR) and Tunable Diode Laser Absorption Spectroscopy (TDLAS) Methods for Determining Stable Isotope Ratios of Atmospheric CO2

    NASA Astrophysics Data System (ADS)

    Ubierna Lopez, N.; Cambaliza, M. L.; Griffith, D. W.; Mount, G. H.; Cousins, A. B.

    2011-12-01

    Worldwide, biosphere-atmosphere carbon exchange and net ecosystem exchange (NEE) are determined using eddy-covariance methods. Information from isotopic CO2 measurements provides valuable constraints to partition NEE into its component fluxes. Stable isotope measurements have traditionally been constrained in frequency by the need to collect and analyze field samples in a laboratory using isotope ratio mass spectrometry (MS). New techniques based on absorption spectroscopy allow for high temporal sampling resolution in the field, but with concerns about precision and accuracy of the isotope-ratios. We tested two absorption spectroscopy systems, a Fourier transformed infrared analyzer (FTIR, Vector 22, Bruker Optics, Ettlingen, Germany) and a tunable diode laser absorption spectrometer (TDLAS, model TGA 100, Campbell Scientific, Inc. Logan, UT, USA), by comparing them with continuous-flow MS (Delta plus XP IRMS, ThermoFinnigan, Bremen, Germany). We conducted a laboratory comparison of gases mixed with various CO2 concentrations and isotopic signatures as well as field-collected samples. The mixed tanks were balanced in ultra-zero air with CO2 concentrations ranging from 353 to 553 ppm, and isotopic compositions (δ13C) between -11.7% to -39.3%. The field samples were collected at four different locations (forest, wheat field, dairy farm, and paper mill) by pumping ambient air into 44- L tanks. Gas from each sample tank was simultaneously delivered to the FTIR and TDLAS systems and subsequently analyzed with continuous-flow MS. The [CO2] determined with the TDLAS or FTIR differed by <1 ppm for CO2-tanks and <2.4 ppm for ambient air samples. The δ13C offset of the CO2 tanks between the MS and the TDLAS and FTIR were on average 0.1% and 0.3%, respectively. However, the offset in TDLAS δ13C values increased for ambient air samples to values of 0.4%, with a maximum of 0.9% for the dairy farm and paper mill samples. Ambient air samples analyzed with the FTIR were on

  17. Cloning of TPS gene from eelgrass species Zostera marina and its functional identification by genetic transformation in rice.

    PubMed

    Zhao, Feng; Li, Qiuying; Weng, Manli; Wang, Xiuliang; Guo, Baotai; Wang, Li; Wang, Wei; Duan, Delin; Wang, Bin

    2013-12-01

    The full-length cDNA sequence (2613 bp) of the trehalose-6-phosphate synthase (TPS) gene of eelgrass Zostera marina (ZmTPS) was identified and cloned. Z. marina is a kind of seed-plant growing in sea water during its whole life history. The open reading frame (ORF) region of ZmTPS gene encodes a protein of 870 amino acid residues and a stop codon. The corresponding genomic DNA sequence is 3770 bp in length, which contains 3 exons and 2 introns. The ZmTPS gene was transformed into rice variety ZH11 via Agrobacterium-mediated transformation method. After antibiotic screening, molecular characterization, salt-tolerance and trehalose content determinations, two transgenic lines resistant to 150 mM NaCL solutions were screened. Our study results indicated that the ZmTPS gene was integrated into the genomic DNA of the two transgenic rice lines and could be expressed well. Moreover, the detection of the transformed ZmTPS gene in the progenies of the two transgenic lines was performed from T1 to T4 generations; and results suggested that the transformed ZmTPS gene can be transmitted from parent to the progeny in transgenic rice. PMID:24035935

  18. AgarTrap-mediated genetic transformation using intact gemmae/gemmalings of the liverwort Marchantia polymorpha L.

    PubMed

    Tsuboyama-Tanaka, Shoko; Kodama, Yutaka

    2015-03-01

    The dioecious liverwort, Marchantia polymorpha L., is an emerging model plant. Various molecular biological techniques have been optimized for M. polymorpha for the past several years, and recently we reported a simplified Agrobacterium-mediated transformation method using sporelings (immature thalli from spores) of M. polymorpha. This method, termed AgarTrap (Agar-utilized Transformation with Pouring Solutions), completed by exchanging appropriate solutions on a single Petri dish to produce a sufficient number of independent transgenic sporelings. However, because spores are produced by crosses between males and females, the genetic backgrounds of resulting transgenic sporelings are not uniform. To easily produce transgenic liverworts with a uniform genetic background using AgarTrap, we developed an AgarTrap-mediated transformation method using intact gemmae/gemmalings produced by asexual reproduction. Using AgarTrap with male and female gemmae/gemmalings produced a sufficient number of independent transgenic gemmalings with uniform genetic backgrounds. The optimized transformation efficiencies were approximately 30 and 50 % in males and females, respectively. As with AgarTrap using sporelings, AgarTrap using intact gemmae/gemmalings will be useful in promoting studies of the molecular biology of M. polymorpha. PMID:25663453

  19. A novel gateway-compatible binary vector series (PC-GW) for flexible cloning of multiple genes for genetic transformation of plants.

    PubMed

    Dalal, Jyoti; Yalamanchili, Roopa; La Hovary, Christophe; Ji, Mikyoung; Rodriguez-Welsh, Maria; Aslett, Denise; Ganapathy, Sowmya; Grunden, Amy; Sederoff, Heike; Qu, Rongda

    2015-09-01

    The rapidly advancing field of plant synthetic biology requires transforming plants with multiple genes. This has sparked a growing interest in flexible plant transformation vectors, which can be used for multi-gene transformations. We have developed a novel binary vector series, named the PC-GW series (GenBank: KP826769-KP826773), for Agrobacterium-mediated plant transformation. The PC-GW vectors use the pCAMBIA vector backbone, and contain NPTII, hpt, bar, mCherry or egfp genes as selectable markers for plant transformation. In a modified multiple cloning site (MCS) of the T-DNA region, we have placed the attR1, attR2 and ccdB sequences for rapid cloning of one to four genes by Gateway™-assisted recombination. In addition, we have introduced four meganuclease sites, and other restriction sites for multi-gene vector construction. Finally, we have placed a CaMV 35S promoter and a 35S terminator on the 5' and 3' ends of the MCS. The CaMV 35S promoter is flanked by PstI restriction sites that can be used to replace it with another promoter sequence if needed. The PC-GW vectors provide choices for selectable markers, cloning methods, and can accommodate up to eight gene constructs in a single T-DNA, thereby significantly reducing the number of transformations or crosses needed to generate multi-transgene expressing plants. PMID:26188330

  20. Stable silylenes.

    PubMed

    Haaf, M; Schmedake, T A; West, R

    2000-10-01

    The field of stable silylene research has grown dramatically since the first isolation of a stable silylene in 1994. Prior to 1994, silylenes existed only as reactive intermediates, isolable only in low-temperature matrixes. Since then, several stable silylenes have been synthesized, some in fact showing remarkable thermal stability. This Account highlights the developments in stable silylene chemistry, including theoretical and experimental studies attempting to explain the remarkable stability of the silylenes as well as the rapidly expanding reaction chemistry of the stable silylenes. PMID:11041835

  1. Carbon transformation and the sources of dissolved inorganic carbonate in sediments of a temperate coastal sea, the Baltic Sea: A stable isotope and modelling approach

    NASA Astrophysics Data System (ADS)

    Lipka, Marko; Liu, Bo; Wegwerth, Antje; Dellwig, Olaf; Winde, Vera; Al-Raei, Abdul M.; Böttcher, Michael E.

    2015-04-01

    Organic matter is mineralized in brackish-marine sediments by microbial activity using predominantly and sulfate as electron acceptors. Under anoxic bottom water conditions, sulfate reduction dominates. Pore water profiles reflect net biogeochemical processes, transformation rates and fluxes of dissolved species across the sediment-water interface. Element fluxes across the sediment-water interface are controlled by different boundary conditions. We present the results of a detailed biogeochemical investigation of interstitial waters from different sediments of the Baltic Sea covering the range of sedimentological and bottom water redox conditions. It was the aim to study the biogeochemical transformation processes and associated element fluxes at the sediment-water-interface and the role of organic matter or methane as potential substrates for microbial activity. Short sediment cores were collected during several research cruises with multicoring devices. Pore waters were analyzed for nutrients, major and trace element concentrations to allow a modelling of net volumetric transformation rates and diffusive element fluxes. Gross sulfate reduction rates were measured in selected cores using incubations with radiotracer. As a tracer for the source of dissolved inorganic carbonate (DIC) the carbon isotope composition was measured. A quantitative interpretation of vertical concentration profiles in the pore waters was performed using different modelling approaches. Element fluxes across the sediment-water interface show a dependence from bottom water redox conditions, sediment compositions, and sedimentation conditions. It is shown that the carbon isotope composition of DIC is a valuable and sensitive parameter in a model-based estimate of the impact of biological and physical mixing of surface sediments. Research is supported by German BMBF within the KÜNO-SECOS project and Leibniz IOW

  2. Mathematical modeling of transformation process of structurally unstable magnetic configurations into structurally stable ones in two-dimensional and three-dimensional geometry

    NASA Astrophysics Data System (ADS)

    Inovenkov, Igor; Echkina, Eugenia; Ponomarenko, Loubov

    Magnetic reconnection is a fundamental process in astrophysical, space and laboratory plasma. In essence, it represents a change of topology of the magnetic field caused by readjustment of the structure of the magnetic field lines. This change leads to release of energy accumulated in the field. We consider transformation process of structurally unstable magnetic configurations into the structurally steady ones from the point of view of the сatastrophe theory. Special attention is paid to modeling of evolution of the structurally unstable three-dimensional magnetic fields.

  3. Stable Expression of mtlD Gene Imparts Multiple Stress Tolerance in Finger Millet

    PubMed Central

    Hema, Ramanna; Vemanna, Ramu S.; Sreeramulu, Shivakumar; Reddy, Chandrasekhara P.; Senthil-Kumar, Muthappa; Udayakumar, Makarla

    2014-01-01

    Finger millet is susceptible to abiotic stresses, especially drought and salinity stress, in the field during seed germination and early stages of seedling development. Therefore developing stress tolerant finger millet plants combating drought, salinity and associated oxidative stress in these two growth stages is important. Cellular protection through osmotic adjustment and efficient free radical scavenging ability during abiotic stress are important components of stress tolerance mechanisms in plants. Mannitol, an osmolyte, is known to scavenge hydroxyl radicals generated during various abiotic stresses and thereby minimize stress damage in several plant species. In this study transgenic finger millet plants expressing the mannitol biosynthetic pathway gene from bacteria, mannitol-1-phosphate dehydrogenase (mtlD), were developed through Agrobacterium tumefaciens-mediated genetic transformation. mtlD gene integration in the putative transgenic plants was confirmed by Southern blot. Further, performance of transgenic finger millet under drought, salinity and oxidative stress was studied at plant level in T1 generation and in T1 and T2 generation seedlings. Results from these experiments showed that transgenic finger millet had better growth under drought and salinity stress compared to wild-type. At plant level, transgenic plants showed better osmotic adjustment and chlorophyll retention under drought stress compared to the wild-type. However, the overall increase in stress tolerance of transgenics for the three stresses, especially for oxidative stress, was only marginal compared to other mtlD gene expressing plant species reported in the literature. Moreover, the Agrobacterium-mediated genetic transformation protocol developed for finger millet in this study can be used to introduce diverse traits of agronomic importance in finger millet. PMID:24922513

  4. Stable transformation of Toxoplasma gondii based on a pyrimethamine resistant trifunctional dihydrofolate reductase-cytosine deaminase-thymidylate synthase gene that confers sensitivity to 5-fluorocytosine.

    PubMed

    Fox, B A; Belperron, A A; Bzik, D J

    1999-01-01

    To improve genetic models available for the analysis of apicomplexan protozoan parasites, bacterial sequences encoding the 427 amino acid cytosine deaminase (CD) gene were fused, in-frame, to an engineered linker domain of the high level pyrimethamine resistant form of the parasite bifunctional dihydrofolate reductase-thymidylate synthase (DHFR-TS) gene. Toxoplasma gondii was transformed with the plasmid containing the fused pyrimethamine resistant dihydrofolate reductase-cytosine deaminase-thymidylate synthase (DHFRm2m3-CD-TS) gene and parasites were selected in a high level of pyrimethamine. Transfected parasites that acquired resistance to pyrimethamine were cloned and evaluated for expression of the CD genetic marker. CD transgenic parasites acquired a high sensitivity to 5-fluorocytosine due to the intraparasitic conversion of this non-toxic prodrug to the cytotoxic compound 5-fluorouracil. Exogenously supplied cytosine or uracil rescued the growth of CD transgenic T. gondii parasites that were cultured in the presence of cytotoxic concentrations of 5-fluorouracil or 5-fluorocytosine. Bacterial CD fused to the pyrimethamine resistant DHFR-TS marker provides a novel genetic tool for new positive and negative genetic selection strategies in several protozoan parasites. An advantage of the CD genetic marker is that it is derived from a bacterial gene and can therefore be used in nearly any parasite genetic background for negative selection. This novel system should facilitate new approaches for the development of improved model genetic systems for the biological investigation of apicomplexan parasites. PMID:10029312

  5. Stable Carbon Isotope Fractionation During 1,2-Dichloropropane-to-Propene Transformation by an Enrichment Culture Containing Dehalogenimonas Strains and a dcpA Gene.

    PubMed

    Martín-González, L; Mortan, S Hatijah; Rosell, M; Parladé, E; Martínez-Alonso, M; Gaju, N; Caminal, G; Adrian, L; Marco-Urrea, E

    2015-07-21

    A stable enrichment culture derived from Besòs river estuary sediments stoichiometrically dechlorinated 1,2-dichloropropane (1,2-DCP) to propene. Sequential transfers in defined anaerobic medium with the inhibitor bromoethanesulfonate produced a sediment-free culture dechlorinating 1,2-DCP in the absence of methanogenesis. Application of previously published genus-specific primers targeting 16S rRNA gene sequences revealed the presence of a Dehalogenimonas strain, and no amplification was obtained with Dehalococcoides-specific primers. The partial sequence of the 16S rRNA amplicon was 100% identical with Dehalogenimonas alkenigignens strain IP3-3. Also, dcpA, a gene described to encode a corrinoid-containing 1,2-DCP reductive dehalogenase was detected. Resistance of the dehalogenating activity to vancomycin, exclusive conversion of vicinally chlorinated alkanes, and tolerance to short-term oxygen exposure is consistent with the hypothesis that a Dehalogenimonas strain is responsible for 1,2-DCP conversion in the culture. Quantitative PCR showed a positive correlation between the number of Dehalogenimonas 16S rRNA genes copies in the culture and consumption of 1,2-DCP. Compound specific isotope analysis revealed that the Dehalogenimonas-catalyzed carbon isotopic fractionation (εC(bulk)) of the 1,2-DCP-to-propene reaction was -15.0 ± 0.7‰ under both methanogenic and nonmethanogenic conditions. This study demonstrates that carbon isotope fractionation is a valuable approach for monitoring in situ 1,2-DCP reductive dechlorination by Dehalogenimonas strains. PMID:26111261

  6. A High-Throughput Regeneration and Transformation Platform for Production of Genetically Modified Banana

    PubMed Central

    Tripathi, Jaindra N.; Oduor, Richard O.; Tripathi, Leena

    2015-01-01

    Banana (Musa spp.) is an important staple food as well as cash crop in tropical and subtropical countries. Various bacterial, fungal, and viral diseases and pests such as nematodes are major constraints in its production and are currently destabilizing the banana production in sub-Saharan Africa. Genetic engineering is a complementary option used for incorporating useful traits in banana to bypass the long generation time, polyploidy, and sterility of most of the cultivated varieties. A robust transformation protocol for farmer preferred varieties is crucial for banana genomics and improvement. A robust and reproducible system for genetic transformation of banana using embryogenic cell suspensions (ECS) has been developed in this study. Two different types of explants (immature male flowers and multiple buds) were tested for their ability to develop ECS in several varieties of banana locally grown in Africa. ECS of banana varieties “Cavendish Williams” and “Gros Michel” were developed using multiple buds, whereas ECS of “Sukali Ndiizi” was developed using immature male flowers. Regeneration efficiency of ECS was about 20,000–50,000 plantlets per ml of settled cell volume (SCV) depending on variety. ECS of three different varieties were transformed through Agrobacterium-mediated transformation using gusA reporter gene and 20–70 independent transgenic events per ml SCV of ECS were regenerated on selective medium. The presence and integration of gusA gene in transgenic plants was confirmed by PCR, dot blot, and Southern blot analysis and expression by histochemical GUS assays. The robust transformation platform was successfully used to generate hundreds of transgenic lines with disease resistance. Such a platform will facilitate the transfer of technologies to national agricultural research systems (NARS) in Africa. PMID:26635849

  7. Agrobacterium tumefaciens-mediated genetic transformation of haptophytes (Isochrysis species).

    PubMed

    Prasad, Binod; Vadakedath, Nithya; Jeong, Hyun-Jeong; General, Thiyam; Cho, Man-Gi; Lein, Wolfgang

    2014-10-01

    Isochrysis galbana and Isochrysis sp. are economically important microalgae from the division of haptophytes. Here, we report Agrobacterium-mediated stable DNA transfer into their nuclear genomes. Initial studies were performed to standardize co-cultivation media and determine the sensitivity of the microalgae to selective agents. Up to 1 mg/ml of the antibiotic hygromycin did not inhibit growth, whereas both the haptophytes bleached in artificial seawater (ASW) medium containing micromolar concentrations of the herbicide norflurazon. Co-cultivation of Isochrysis sp. and I. galbana with Agrobacterium tumefaciens strain LBA 4404 harboring the binary vector pCAMBIA 1380-pds-L504R yielded norflurazon-resistant (NR) colonies visible on selective plates after 20-30 days. pCAMBIA 1380-pds-L540R was constructed by cloning a mutated genomic phytoene desaturase (pds) gene from Haematococcus pluvialis as a selectable marker gene into the binary vector system pCAMBIA 1380. Co-cultivation of Isochrysis sp. with A. tumefaciens in ASW medium containing 200 μM of acetosyringone for 72 h produced the highest number of NR cells. For I. galbana, 100 μM of acetosyringone, ASW medium, and 48 h co-cultivation period appeared to be optimum co-cultivation parameters. The NR colonies kept their resistance phenotype for at least 24 months, even in the absence of selective pressure. The transfer of the pds gene in NR cells was shown by PCR amplification of the T-DNA sequences from the genomic DNA of NR cells and Southern blot analysis using T-DNA sequences as probes. The genetic manipulation described here will allow metabolic engineering and a better understanding of several biochemical pathways in the future. PMID:24993358

  8. Co-transforming bar and CsLEA enhanced tolerance to drought and salt stress in transgenic alfalfa (Medicago sativa L.).

    PubMed

    Zhang, Jiyu; Duan, Zhen; Zhang, Daiyu; Zhang, Jianquan; Di, Hongyan; Wu, Fan; Wang, Yanrong

    2016-03-25

    Drought and high salinity are two major abiotic factors that restrict alfalfa productivity. A dehydrin protein, CsLEA, from the desert grass Cleistogenes songorica was transformed into alfalfa (Medicago sativa L.) via Agrobacterium-mediated transformation using the bar gene as a selectable marker, and the drought and salt stress tolerances of the transgenic plants were assessed. Thirty-nine of 119 transformants were positive, as screened by Basta, and further molecularly authenticated using PCR and RT-PCR. Phenotype observations revealed that the transgenic plants grew better than the wild-type (WT) plants after 15d of drought stress and 10d of salt stress: the leaves of WT alfalfa turned yellow, whereas the transgenic alfalfa leaves only wilted; after rewatering, the transgenic plants returned to a normal state, though the WT plants could not be restored. Evaluation of physiologic and biochemical indices during drought and salt stresses showed a relatively lower Na(+) content in the leaves of the transgenic plants, which would reduce toxic ion effects. In addition, the transgenic plants were able to maintain a higher relative water content (RWC), higher shoot biomass, fewer photosystem changes, decreased membrane injury, and a lower level of osmotic stress injury. These results demonstrate that overexpression of the CsLEA gene can enhance the drought and salt tolerance of transgenic alfalfa; in addition, carrying the bar gene in the genome may increase herbicide resistance. PMID:26906624

  9. Stable compactifications

    SciTech Connect

    Accetta, F.S.; Gleiser, M.; Holman, R.; Kolb, E.W.

    1986-03-01

    We show that compactifications of theories with extra dimensions are unstable if due to monopole configurations of an antisymmetric tensor field balanced against one-loop Casimir corrections. In the case of ten dimensional supergravity, it is possible, at least for a portion of the phase space, to achieve a stable compactification without fine-tuning by including the contribution of fermionic condensates to the monopole configurations. 23 refs., 2 figs.

  10. Unlocking the potential of tropical root crop biotechnology in east Africa by establishing a genetic transformation platform for local farmer-preferred cassava cultivars

    PubMed Central

    Nyaboga, Evans; Njiru, Joshua; Nguu, Edward; Gruissem, Wilhelm; Vanderschuren, Herve; Tripathi, Leena

    2013-01-01

    Cassava genetic transformation capacity is still mostly restricted to advanced laboratories in the USA, Europe and China; and its implementation and maintenance in African laboratories has remained scarce. The impact of transgenic technologies for genetic improvement of cassava will depend largely on the transfer of such capabilities to researchers in Africa, where cassava has an important socioeconomic niche. A major constraint to the development of genetic transformation technologies for cassava improvement has been the lack of an efficient and robust transformation and regeneration system. Despite the success achieved in genetic modification of few cassava cultivars, including the model cultivar 60444, transgenic cassava production remains difficult for farmer-preferred cultivars. In this study, a protocol for cultivar 60444 developed at ETH Zurich was successfully implemented and optimized to establish transformation of farmer-preferred cassava cultivars popular in east Africa. The conditions for production and proliferation of friable embryogenic calli (FEC) and Agrobacterium-mediated transformation were optimized for three east African farmer-preferred cultivars (Ebwanatereka, Kibandameno and Serere). Our results demonstrated transformation efficiencies of about 14–22 independent transgenic lines per 100 mg of FEC for farmer-preferred cultivars in comparison to 28 lines per 100 mg of the model cultivar 60444. The presence, integration and expression of the transgenes were confirmed by PCR, Southern blot analysis and histochemical GUS assay. This study reports the establishment of a cassava transformation platform at International Institute of Tropical Agriculture (IITA) hosted by Biosciences eastern and central Africa (BecA) hub in Kenya and provides the basis for transferring important traits such as virus resistance and prolonged shelf-life to farmer-preferred cultivars in east Africa. We anticipate that such platform will also be instrumental to transfer

  11. Development of an efficient transformation method by Agrobacterium tumefaciens and high throughput spray assay to identify transgenic plants for woodland strawberry (Fragaria vesca) using NPTII selection.

    PubMed

    Pantazis, Christopher J; Fisk, Sarah; Mills, Kerri; Flinn, Barry S; Shulaev, Vladimir; Veilleux, Richard E; Dan, Yinghui

    2013-03-01

    KEY MESSAGE : We developed an efficient Agrobacterium -mediated transformation method using an Ac/Ds transposon tagging construct for F. vesca and high throughput paromomycin spray assay to identify its transformants for strawberry functional genomics. Genomic resources for Rosaceae species are now readily available, including the Fragaria vesca genome, EST sequences, markers, linkage maps, and physical maps. The Rosaceae Genomic Executive Committee has promoted strawberry as a translational genomics model due to its unique biological features and transformability for fruit trait improvement. Our overall research goal is to use functional genomic and metabolic approaches to pursue high throughput gene discovery in the diploid woodland strawberry. F. vesca offers several advantages of a fleshy fruit typical of most fruit crops, short life cycle (seed to seed in 12-16 weeks), small genome size (206 Mbb/C), small plant size, self-compatibility, and many seeds per plant. We have developed an efficient Agrobacterium tumefaciens-mediated strawberry transformation method using kanamycin selection, and high throughput paromomycin spray assay to efficiently identify transgenic strawberry plants. Using our kanamycin transformation method, we were able to produce up to 98 independent kanamycin resistant insertional mutant lines using a T-DNA construct carrying an Ac/Ds transposon Launchpad system from a single transformation experiment involving inoculation of 22 leaf explants of F. vesca accession 551572 within approx. 11 weeks (from inoculation to soil). Transgenic plants with 1-2 copies of a transgene were confirmed by Southern blot analysis. Using our paromomycin spray assay, transgenic F. vesca plants were rapidly identified within 10 days after spraying. PMID:23160638

  12. High efficiency plum (Prunus domestica L.) transformation: a functional genomics tool for Rosaceae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have developed an improved Agrobacterium-mediated protocol in plum (Prunus domestica L.) through the addition of 2,4-D to the regeneration media. This method has increased the regeneration efficiency of independent transgenic plants up to 10-fold over previous reports. DNA blot analysis of puta...

  13. In planta Transformed Cumin (Cuminum cyminum L.) Plants, Overexpressing the SbNHX1 Gene Showed Enhanced Salt Endurance.

    PubMed

    Pandey, Sonika; Patel, Manish Kumar; Mishra, Avinash; Jha, Bhavanath

    2016-01-01

    Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas. PMID:27411057

  14. In planta Transformed Cumin (Cuminum cyminum L.) Plants, Overexpressing the SbNHX1 Gene Showed Enhanced Salt Endurance

    PubMed Central

    Pandey, Sonika; Patel, Manish Kumar; Jha, Bhavanath

    2016-01-01

    Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas. PMID:27411057

  15. pSiM24 is a novel versatile gene expression vector for transient assays as well as stable expression of foreign genes in plants.

    PubMed

    Sahoo, Dipak Kumar; Dey, Nrisingha; Maiti, Indu Bhushan

    2014-01-01

    We have constructed a small and highly efficient binary Ti vector pSiM24 for plant transformation with maximum efficacy. In the pSiM24 vector, the size of the backbone of the early binary vector pKYLXM24 (GenBank Accession No. HM036220; a derivative of pKYLX71) was reduced from 12.8 kb to 7.1 kb. The binary vector pSiM24 is composed of the following genetic elements: left and right T-DNA borders, a modified full-length transcript promoter (M24) of Mirabilis mosaic virus with duplicated enhancer domains, three multiple cloning sites, a 3'rbcsE9 terminator, replication functions for Escherichia coli (ColE1) and Agrobacterium tumefaciens (pRK2-OriV) and the replicase trfA gene, selectable marker genes for kanamycin resistance (nptII) and ampicillin resistance (bla). The pSiM24 plasmid offers a wide selection of cloning sites, high copy numbers in E. coli and a high cloning capacity for easily manipulating different genetic elements. It has been fully tested in transferring transgenes such as green fluorescent protein (GFP) and β-glucuronidase (GUS) both transiently (agro-infiltration, protoplast electroporation and biolistic) and stably in plant systems (Arabidopsis and tobacco) using both agrobacterium-mediated transformation and biolistic procedures. Not only reporter genes, several other introduced genes were also effectively expressed using pSiM24 expression vector. Hence, the pSiM24 vector would be useful for various plant biotechnological applications. In addition, the pSiM24 plasmid can act as a platform for other applications, such as gene expression studies and different promoter expressional analyses. PMID:24897541

  16. pSiM24 Is a Novel Versatile Gene Expression Vector for Transient Assays As Well As Stable Expression of Foreign Genes in Plants

    PubMed Central

    Sahoo, Dipak Kumar; Dey, Nrisingha; Maiti, Indu Bhushan

    2014-01-01

    We have constructed a small and highly efficient binary Ti vector pSiM24 for plant transformation with maximum efficacy. In the pSiM24 vector, the size of the backbone of the early binary vector pKYLXM24 (GenBank Accession No. HM036220; a derivative of pKYLX71) was reduced from 12.8 kb to 7.1 kb. The binary vector pSiM24 is composed of the following genetic elements: left and right T-DNA borders, a modified full-length transcript promoter (M24) of Mirabilis mosaic virus with duplicated enhancer domains, three multiple cloning sites, a 3′rbcsE9 terminator, replication functions for Escherichia coli (ColE1) and Agrobacterium tumefaciens (pRK2-OriV) and the replicase trfA gene, selectable marker genes for kanamycin resistance (nptII) and ampicillin resistance (bla). The pSiM24 plasmid offers a wide selection of cloning sites, high copy numbers in E. coli and a high cloning capacity for easily manipulating different genetic elements. It has been fully tested in transferring transgenes such as green fluorescent protein (GFP) and β-glucuronidase (GUS) both transiently (agro-infiltration, protoplast electroporation and biolistic) and stably in plant systems (Arabidopsis and tobacco) using both agrobacterium-mediated transformation and biolistic procedures. Not only reporter genes, several other introduced genes were also effectively expressed using pSiM24 expression vector. Hence, the pSiM24 vector would be useful for various plant biotechnological applications. In addition, the pSiM24 plasmid can act as a platform for other applications, such as gene expression studies and different promoter expressional analyses. PMID:24897541

  17. Transformation pathways of liposomes.

    PubMed

    Hotani, H

    1984-09-01

    Liposomes undergoing transformation were observed by dark-field light microscopy in order to study the role of lipid in morphogenesis of biological vesicular structures. Liposomes were found to transform sequentially in a well-defined manner through one of several transformation pathways. A circular biconcave form was an initial shape in all the pathways and it transformed into a stable thin flexible filament or small spheres via a variety of regularly shaped vesicles which possessed geometrical symmetry. The transformation was reversible up to a certain point in each pathway. Osmotic pressure was found to be the driving force for the transformations. Biological membrane vesicles such as trypsinized red cell ghosts also transformed by similar pathways. PMID:6548263

  18. Stable isotope ratios of nonexchangeable hydrogen in organic matter of soils and plants along a 2100-km climosequence in Argentina: New insights into soil organic matter sources and transformations?

    NASA Astrophysics Data System (ADS)

    Ruppenthal, Marc; Oelmann, Yvonne; del Valle, Héctor Francisco; Wilcke, Wolfgang

    2015-03-01

    The quantitative contribution of shoot and root organic matter (OM) to the soil carbon (C) stock is still unknown, mainly because of methodological restrictions. The novel measurement of the nonexchangeable hydrogen (H) stable isotope ratio (δ2Hn) in bulk OM provides new opportunities to investigate the sources of soil C and its climate-dependent transformations. Our objectives were to test whether (I) there are systematic differences between δ2Hn values of aboveground and belowground plant OM and (II) δ2Hn values of litter and soil OM relate to climate and plant OM source δ2Hn signals. We determined δ2Hn values of bulk shoot, root, litter and demineralized soil OM from 20 sampling sites along a 2100-km climosequence from the Argentinean Pampas to the Patagonian steppe. The δ2Hn values of shoot and litter OM correlated closely with the aridity index (r = -0.83, p < 0.001 and r = -0.78, p < 0.001, respectively) because of H isotope fractionation during aridity-controlled transpiration in shoots. In contrast, δ2Hn values of root and soil OM showed a close correlation with modeled mean annual δ2H values of local precipitation (r = 0.91, p < 0.001 and r = 0.97, p < 0.001, respectively, root mean square error of 8.2‰ and 7.2‰, respectively) and thus mean annual temperature (r = 0.80, p < 0.001 and r = 0.88, p < 0.001, respectively). δ2Hn values of shoot and root OM differed markedly (no linear correlation) most likely because of biosynthetic exchange of C-bound H with ambient water in the roots. δ2Hn values of root and demineralized soil OM, however, were closely correlated (r = 0.91, p < 0.001) with a constant offset irrespective of climatic conditions, suggesting that root OM was a more important source of soil OM than shoot OM. A possible contribution of shoot OM to soil OM could only be explained if shoot OM underwent biosynthetic exchange of C-bound H with ambient water in soil during microbial and fungal decomposition. This mechanism is known for

  19. Agrobacterium-mediated inoculation of chrysanthemum (Chrysanthemum morifolium) plants with chrysanthemum stunt viroid.

    PubMed

    Nabeshima, Tomoyuki; Doi, Motoaki; Hosokawa, Munetaka

    2016-08-01

    Agroinfiltration was tested as a method of inoculation of chrysanthemum plants with chrysanthemum stunt viroid (CSVd). Binary vectors harboring dimeric CSVd sequences in sense and antisense orientations were constructed, and Agrobacterium transfected with these binary vectors was infiltrated into chrysanthemum leaves. Northern blotting and reverse transcription polymerase chain reaction analysis showed that local infection was established within 7 days and systemic infection within 20 days. CSVd polarities showed no difference in infectivity. This study showed that agroinfiltration of chrysanthemum plants is an easy, rapid, and cost-effective method for CSVd inoculation. PMID:27155239

  20. Potassium chloride and rare earth elements improve plant growth and increase the frequency of the Agrobacterium tumefaciens-mediated plant transformation.

    PubMed

    Boyko, Alex; Matsuoka, Aki; Kovalchuk, Igor

    2011-04-01

    Plant transformation efficiency depends on the ability of the transgene to successfully interact with plant host factors. Our previous work and the work of others showed that manipulation of the activity of host factors allows for increased frequency of transformation. Recently we reported that exposure of tobacco plants to increased concentrations of ammonium nitrate increases the frequency of both homologous recombination and plant transgenesis. Here we tested the influence of KCl and salts of rare earth elements, Ce and La on the efficiency of Agrobacterium-mediated plant transformation. We found that exposure to KCl, CeCl(3) and LaCl(3) leads to an increase in recombination frequency in Arabidopsis and tobacco. Plants grown in the presence of CeCl(3) and LaCl(3) had higher biomass, longer roots and greater root number. Analysis of transformation efficiency showed that exposure of tobacco plants to 50 mM KCl resulted in ~6.0-fold increase in the number of regenerated calli and transgenic plants as compared to control plants. Exposure to various concentrations of CeCl(3) showed a maximum increase of ~3.0-fold in both the number of calli and transgenic plants. Segregation analysis showed that exposure to KCl and cerium (III) chloride leads to more frequent integrations of the transgene at a single locus. Analysis of transgene intactness showed better preservation of right T-DNA border during transgene integration. Our data suggest that KCl and CeCl(3) can be effectively used to improve quantity and quality of transgene integrations. PMID:21132499

  1. Transformation and Evaluation of Cry1Ac+Cry2A and GTGene in Gossypium hirsutum L.

    PubMed Central

    Puspito, Agung N.; Rao, Abdul Q.; Hafeez, Muhammad N.; Iqbal, Muhammad S.; Bajwa, Kamran S.; Ali, Qurban; Rashid, Bushra; Abbas, Muhammad A.; Latif, Ayesha; Shahid, Ahmad A.; Nasir, Idrees A.; Husnain, Tayyab

    2015-01-01

    More than 50 countries around the globe cultivate cotton on a large scale. It is a major cash crop of Pakistan and is considered “white gold” because it is highly important to the economy of Pakistan. In addition to its importance, cotton cultivation faces several problems, such as insect pests, weeds, and viruses. In the past, insects have been controlled by insecticides, but this method caused a severe loss to the economy. However, conventional breeding methods have provided considerable breakthroughs in the improvement of cotton, but it also has several limitations. In comparison with conventional methods, biotechnology has the potential to create genetically modified plants that are environmentally safe and economically viable. In this study, a local cotton variety VH 289 was transformed with two Bt genes (Cry1Ac and Cry2A) and a herbicide resistant gene (cp4 EPSPS) using the Agrobacterium mediated transformation method. The constitutive CaMV 35S promoter was attached to the genes taken from Bacillus thuringiensis (Bt) and to an herbicide resistant gene during cloning, and this promoter was used for the expression of the genes in cotton plants. This construct was used to develop the Glyphosate Tolerance Gene (GTGene) for herbicide tolerance and insecticidal gene (Cry1Ac and Cry2A) for insect tolerance in the cotton variety VH 289. The transgenic cotton variety performed 85% better compared with the non-transgenic variety. The study results suggest that farmers should use the transgenic cotton variety for general cultivation to improve the production of cotton. PMID:26617613

  2. A high-throughput transformation system in plum (Prunus domestica L.) provides a powerful tool for functional genomics in Rosaceae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An improved Agrobacterium-mediated protocol in plum (Prunus domestica L.) is described, whereby, the addition of 2,4-D to the regeneration media increased the regeneration efficiency of independent transgenic plants up to 10 fold over previous reports. DNA blot analysis of putative transgenic shoot...

  3. A high-throughput transformation system in plum (Prunus domestica L.) for functional genomics research in Rosaceae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have developed an improved Agrobacterium-mediated protocol in plum (Prunus domestica L.) through the addition of 2, 4-D to the regeneration media. This method has increased the regeneration efficiency of independent transgenic plants up to 10 fold over previous reports. DNA blot analysis of put...

  4. A high-throughput transformation system in plum (Prunus domestica L.) useful for functional genomics in Rosaceae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have developed an improved Agrobacterium-mediated protocol in plum (Prunus domestica L.) through the addition of 2,4-D to the regeneration media. This method has increased the regeneration efficiency of independent transgenic plants up to 10 fold over previous reports. DNA blot analysis of puta...

  5. Co-transforming bar and CsALDH Genes Enhanced Resistance to Herbicide and Drought and Salt Stress in Transgenic Alfalfa (Medicago sativa L.)

    PubMed Central

    Duan, Zhen; Zhang, Daiyu; Zhang, Jianquan; Di, Hongyan; Wu, Fan; Hu, Xiaowen; Meng, Xuanchen; Luo, Kai; Zhang, Jiyu; Wang, Yanrong

    2015-01-01

    Drought and high salinity are two major abiotic factors that restrict the productivity of alfalfa. By application of the Agrobacterium-mediated transformation method, an oxidative responsive gene, CsALDH12A1, from the desert grass Cleistogenes songorica together with the bar gene associated with herbicide resistance, were co-transformed into alfalfa (Medicago sativa L.). From the all 90 transformants, 16 were positive as screened by spraying 1 mL L-1 10% Basta solution and molecularly diagnosis using PCR. Real-time PCR analysis indicated that drought and salt stress induced high CsALDH expression in the leaves of the transgenic plants. The CsALDH expression levels under drought (15 d) and salt stress (200 mM NaCl) were 6.11 and 6.87 times higher than in the control plants, respectively. In comparison to the WT plants, no abnormal phenotypes were observed among the transgenic plants, which showed significant enhancement of tolerance to 15 d of drought and 10 d of salinity treatment. Evaluation of the physiological and biochemical indices during drought and salt stress of the transgenic plants revealed relatively lower Na+ content and higher K+ content in the leaves relative to the WT plants, a reduction of toxic on effects and maintenance of osmotic adjustment. In addition, the transgenic plants could maintain a higher relative water content level, higher shoot biomass, fewer changes in the photosystem, decreased membrane injury, and a lower level of osmotic stress. These results indicate that the co-expression of the introduced bar and CsALDH genes enhanced the herbicide, drought and salt tolerance of alfalfa and therefore can potentially be used as a novel genetic resource for the future breeding programs to develop new cultivars. PMID:26734025

  6. Clovers (Trifolium spp.).

    PubMed

    Rahimi-Ashtiani, Samira; Sahab, Sareena; Panter, Stephen; Mason, John; Spangenberg, German

    2015-01-01

    Clovers (Trifolium spp.) constitute one of the major forage legumes widely grown for its rich protein content and its major role in maintaining environmental sustainability by improving the soil fertility. Gene technology can assist plant improvement efforts in clovers (Trifolium spp.), aiming to improve forage quality, yield, and adaptation to biotic and abiotic stresses. An efficient and reproducible protocol for Agrobacterium-mediated transformation of a range of Trifolium species, using cotyledonary explants and different selectable marker genes, is described. The protocol is robust and allows for genotype and Agrobacterium strain-independent transformation of clovers. Stable meiotic transmission of transgenes has been demonstrated for selected transgenic clovers carrying single T-DNA inserts recovered from Agrobacterium-mediated transformation. This methodology can also be successfully used for "isogenic transformation" in clovers: the generation of otherwise identical plants with and without the transgene from the two cotyledons of a single seed. Stable transgenes may be used in further functional genomics, develop new traits and profile gene expression using reporters, and facilitate purification of tissue or single cells. PMID:25300844

  7. Genetic transformation and expression of transgenic lines of Populus x euramericana with insect-resistance and salt-tolerance genes.

    PubMed

    Yang, R L; Wang, A X; Zhang, J; Dong, Y; Yang, M S; Wang, J M

    2016-01-01

    We characterized new transgenic varieties of poplar with multiple insect-resistant and salt stress tolerant genes. Two insect-resistant Bacillus thuringiensis (Bt) genes, Cry1Ac and Cry3A, and a salt-tolerant gene, Betaine aldehyde dehydrogenase (BADH) were inserted into a vector, p209-Cry1Ac-Cry3A-BADH. The clone of Populus x euramericana was transformed by the vector using the Agrobacterium-mediated method. Three transgenic lines were assessed using genetic detection and resistance expression analysis. PCR revealed that exogenous genes Cry1Ac, Cry3A, BADH and selective marker gene NPTII were present in three transgenic lines. Quantitative real-time PCR (qPCR) showed significant differences in the transcriptional abundance of three exogenous genes in different lines. Results of assays for Bt toxic proteins showed that the Cry1Ac and Cry3A toxic protein content of each line was 12.83-26.32 and 2108.91-2724.79 ng/g, respectively. The Cry1Ac toxic protein content of different lines was significantly different; the Cry3A toxic protein content was about 100 times higher than that of the Cry1Ac toxic protein. The insect-resistance test revealed the mortality rate of transgenic lines to Hyphantria cunea L1 larvae varied by 42.2-66.7%, which was significantly higher than non-transgenic lines. The mortality rate of L1 and L2 Plagiodera versicolora larvae was 100%. The insecticidal effect of transgenic lines to P. versicolora larvae was higher than that to H. cunea larvae. NaCl stress tolerance of three transgenic lines under 3-6% NaCl concentration was significantly higher than that of non-transgenic lines. PMID:27173305

  8. Stable Fly Research

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Adult stable flies feed on the blood of humans, pets and livestock, inflicting painful bites. Stable flies need one and sometimes two bloodmeals each day to develop their eggs. Unlike mosquitoes where only the females bloodfeed, both male and female stable flies require blood to reproduce. Stable fl...

  9. Cereal transformation through particle bombardment

    NASA Technical Reports Server (NTRS)

    Casas, A. M.; Kononowicz, A. K.; Bressan, R. A.; Hasegawa, P. M.; Mitchell, C. A. (Principal Investigator)

    1995-01-01

    The review focuses on experiments that lead to stable transformation in cereals using microprojectile bombardment. The discussion of biological factors that affect transformation examines target tissues and vector systems for gene transfer. The vector systems include reporter genes, selectable markers, genes of agronomic interest, and vector constructions. Other topics include physical parameters that affect DNA delivery, selection of stably transformed cells and plant regeneration, and analysis of gene expression and transmission to the progeny.

  10. Magnetotelluric Data, Stable Distributions and Stable Regression

    NASA Astrophysics Data System (ADS)

    Chave, A. D.

    2013-12-01

    The author has noted for many years that the residuals from robust or bounded influence estimates of the magnetotelluric response function are systematically long tailed compared to a Gaussian or Rayleigh distribution. Consequently, the standard statistical model of a Gaussian core contaminated by a fraction of outlying data is not really valid. However, the typical result is an improvement on ordinary least squares, and has become standard in the electromagnetic induction community. A recent re-evaluation of the statistics of magnetotelluric response function estimation has shown that, in almost all cases, the residuals are alpha stable rather than Gaussian. Alpha stable distributions are characterized by four parameters: a shape parameter lying on (0, 2], a skewness parameter, a scale parameter and a location parameter, and cannot be expressed in closed form except for a few special cases. When the shape parameter is 2, the result is Gaussian, but when it is smaller the resulting distribution has infinite variance. Typical magnetotelluric residuals are alpha stable with a shape parameter lying between 1 and 2. This suggests that robust methods improve response function estimates by eliminating data corresponding to the largest stable residuals while leaving the bulk of the population alone. A better statistical approach is based on stable regression that directly accommodates the actual residual distribution without eliminating the most extreme ones. This paper will introduce such an algorithm, and illustrate its functionality with a variety of magnetotelluric data. Further work remains to produce a robust stable regression algorithm that will eliminate real outliers such as lightning strikes or instrument problems without affecting the bulk stable population. Stable distributions are intimately associated with fractional derivative physical processes. Since the Maxwell equations and the constitutive relations pertaining to the earth do not contain any fractional

  11. Crystal transformation synthesis of a highly stable fluorescent 3D indium-tetranuclear {In4(μ2-OH)3} building block based metal organic framework through a dinuclear complex.

    PubMed

    Wang, Xin-Ming; Fan, Rui-Qing; Qiang, Liang-Sheng; Wang, Ping; Yang, Yu-Lin; Wang, Yu-Lei

    2014-11-21

    A rare 3D tetranuclear {In4(μ2-OH)3} building block based MOF {[In4/3(μ2-OH)(2,6-pydc)(1,4-bda)0.5(H2O)]·2H2O}n (2) was obtained through a crystal transformation from a dimeric complex In3(2,6-pydc)3(1,4-bda)1.5(H2O)6 (1). With a 2D + 3D → 3D compact structure, 2 retains crystallinity in boiling water and organic solvents, exhibiting exceptional fluorescence quenching behaviour for the DMSO molecule. PMID:25135576

  12. Staff detection with stable paths.

    PubMed

    Dos Santos Cardoso, Jaime; Capela, Artur; Rebelo, Ana; Guedes, Carlos; Pinto da Costa, Joaquim

    2009-06-01

    The preservation of musical works produced in the past requires their digitalization and transformation into a machine-readable format. The processing of handwritten musical scores by computers remains far from ideal. One of the fundamental stages to carry out this task is the staff line detection. We investigate a general-purpose, knowledge-free method for the automatic detection of music staff lines based on a stable path approach. Lines affected by curvature, discontinuities, and inclination are robustly detected. Experimental results show that the proposed technique consistently outperforms well-established algorithms. PMID:19372615

  13. Angina Pectoris (Stable Angina)

    MedlinePlus

    ... Pressure High Blood Pressure Tools & Resources Stroke More Angina Pectoris (Stable Angina) Updated:Aug 24,2016 You may have heard the term “angina pectoris” or “stable angina” in your doctor’s office, but ...

  14. Reversible transformation of a stable monomeric silicon(II) compound into a stable disilene by phase transfer: experimental and theoretical studies of the system {[(Me3Si)2N](Me5C5)Si}n with n = 1,2.

    PubMed

    Jutzi, Peter; Mix, Andreas; Neumann, Beate; Rummel, Britta; Schoeller, Wolfgang W; Stammler, Hans-Georg; Rozhenko, A B

    2009-09-01

    The salt (eta(5)-pentamethylcyclopentadienyl)silicon(II) tetrakis(pentafluorophenyl)borate (5) reacts at -78 degrees C with lithium bis(trimethylsilyl)amide in dimethoxyethane (DME) as solvent to give quantitatively the compound [bis(trimethylsilyl)amino][pentamethylcyclopentadienyl]silicon(II) 6A in the form of a colorless viscous oil. The reaction performed at -40 degrees C leads to the silicon(IV) compound 7, the formal oxidative addition product of 6A with DME. Cycloaddition is observed in the reaction of 6A with 2,3-dimethylbutadiene to give the silicon(IV) compound 8. Upon attempts to crystallize 6A from organic solvents such as hexane, THF, or toluene, the deep yellow compound trans-1,2-bis[bis(trimethylsilyl)amino]-1,2-bis(pentamethylcyclopentadienyl)disilene (6B), the formal dimer of 6A, crystallizes from the colorless solution, but only after several days or even weeks. Upon attempts to dissolve the disilene 6B in the described organic solvents, a colorless solution is obtained after prolonged vigorous shaking or ultrasound treatment. From this solution, pure 6A can be recovered after solvent evaporation. This transformation process can be repeated several times. In a mass spectroscopic investigation of 6B, Si=Si bond cleavage is observed to give the molecular ion with the composition of 6A as the fragment with the highest mass. The X-ray crystal structure analysis of the disilene 6B supports a molecule with a short Si=Si bond (2.168 A) with efficiently packed, rigid sigma-bonded cyclopentadienyl substituents and silylamino groups. The conformation of the latter does not allow electron donation to the central silicon atom. Theoretical calculations at the density functional level (RI-BP86 and B3LYP, TZVP basis set) confirm the structure of 6B and reveal for silylene 6A the presence of an eta(2)-bonded cyclopentadienyl ligand and of a silylamino group in a conformation that prevents electron back-donation. Further theoretical calculations for the silicon

  15. Reading Transformation

    ERIC Educational Resources Information Center

    Reeves, Melinda

    2006-01-01

    The parents of students who attend Decatur High School thought that there was little hope of their kids going on to college. After a year or so in Decatur's reading program, their sons and daughters were both transformed and college bound. In this article, the author describes how Decatur was able to successfully transform their students. Seven…

  16. Apricot (Prunus armeniaca L.).

    PubMed

    Petri, César; Alburquerque, Nuria; Burgos, Lorenzo

    2015-01-01

    A protocol for Agrobacterium-mediated stable transformation of whole leaf explants of the apricot (Prunus armeniaca) cultivars 'Helena' and 'Canino' is described. Regenerated buds were selected using a two-step selection strategy with paromomycin sulfate and transferred to bud multiplication medium 1 week after they were detected for optimal survival. After buds were transferred to bud multiplication medium, antibiotic was changed to kanamycin and concentration increased gradually at each transfer to fresh medium in order to eliminate possible escapes and chimeras. Transformation efficiency, based on PCR analysis of individual putative transformed shoots from independent lines, was 5.6%. Green and healthy buds, surviving high kanamycin concentration, were transferred to shoot multiplication medium where they elongated in shoots and proliferated. Elongated transgenic shoots were rooted in a medium containing 70 μM kanamycin. Rooted plants were acclimatized following standard procedures. This constitutes the only transformation protocol described for apricot clonal tissues and one of the few of Prunus. PMID:25416253

  17. Electronically-Scanned Fourier-Transform Spectrometer

    NASA Technical Reports Server (NTRS)

    Breckinridge, J. B.; Ocallaghan, F. G.

    1984-01-01

    Instrument efficient, lightweight, and stable. Fourier-transform spectrometer configuration uses electronic, instead of mechanical, scanning. Configuration insensitive to vibration-induced sampling errors introduced into mechanically scanned systems.

  18. Lightweight transformer

    SciTech Connect

    Swallom, D.W.; Enos, G.

    1990-05-01

    The technical effort described in this report relates to the program that was performed to design, fabricate, and test a lightweight transformer for Strategic Defense Initiative Organization (SDIO) mission requirements. The objectives of this program were two-fold: (1) design and fabricate a lightweight transformer using liquid hydrogen as the coolant; and (2) test the completed transformer assembly with a low voltage, dc power source. Although the full power testing with liquid helium was not completed, the program demonstrated the viability of the design approach. The lightweight transformer was designed and fabricated, and low and moderate power testing was completed. The transformer is a liquid hydrogen cooled air core transformer that uses thin copper for its primary and secondary windings. The winding mass was approximately 12 kg, or 0.03 kg/kW. Further refinements of the design to a partial air core transformer could potentially reduce the winding mass to as low as 4 or 5 kg, or 0.0125 kg/kW. No attempt was made on this program to reduce the mass of the related structural components or cryogenic container. 8 refs., 39 figs., 2 tabs.

  19. Triple transformation

    NASA Astrophysics Data System (ADS)

    Khan, Farrukh I.; Schinn, Dustin S.

    2013-08-01

    A new business plan that enables policy transformation and resource mobilization at the national and international level, while improving access to resources, will allow the Green Climate Fund to integrate development goals and action on climate change.

  20. Covariant Transform

    NASA Astrophysics Data System (ADS)

    Kisil, Vladimir V.

    2011-03-01

    Dedicated to the memory of Cora Sadosky The paper develops theory of covariant transform, which is inspired by the wavelet construction. It was observed that many interesting types of wavelets (or coherent states) arise from group representations which are not square integrable or vacuum vectors which are not admissible. Covariant transform extends an applicability of the popular wavelets construction to classic examples like the Hardy space H2, Banach spaces, covariant functional calculus and many others.

  1. Stable growth transformation of human T lymphocytes by herpesvirus saimiri.

    PubMed Central

    Biesinger, B; Müller-Fleckenstein, I; Simmer, B; Lang, G; Wittmann, S; Platzer, E; Desrosiers, R C; Fleckenstein, B

    1992-01-01

    Herpesvirus saimiri induces T-cell lymphomas in various species of New World monkeys and in rabbits, and it is able to immortalize monkey T lymphocytes in vitro. Sequences responsible for these effects have been localized to a region of the genome that varies significantly among the virus subgroups A, B, and C. We now report that infection of human blood lymphocytes and thymocytes with strains of subgroup C, in contrast to viruses of the other subgroups, yields continuously proliferating T-cell lines with the phenotype of mature CD4- or CD8-positive cells. Infection with strains of Herpes-virus saimiri subgroup C can thus be used to generate human T-cell lines for a variety of immunological and developmental studies. Images PMID:1313581

  2. FRACTIONATION OF STABLE CARBON ISOTOPES DURING ABIOTIC TRANSFORMATION OF TCE

    EPA Science Inventory

    At a Superfund Site in Minnesota, ground water is contaminated with trichloroethylene (TCE) with the contaminant plume stretching over five miles long. The ground water is iron and manganese reducing, and the complete absence of dichloroethylene, vinyl chloride, and ethene in th...

  3. Deformed Palmprint Matching Based on Stable Regions.

    PubMed

    Wu, Xiangqian; Zhao, Qiushi

    2015-12-01

    Palmprint recognition (PR) is an effective technology for personal recognition. A main problem, which deteriorates the performance of PR, is the deformations of palmprint images. This problem becomes more severe on contactless occasions, in which images are acquired without any guiding mechanisms, and hence critically limits the applications of PR. To solve the deformation problems, in this paper, a model for non-linearly deformed palmprint matching is derived by approximating non-linear deformed palmprint images with piecewise-linear deformed stable regions. Based on this model, a novel approach for deformed palmprint matching, named key point-based block growing (KPBG), is proposed. In KPBG, an iterative M-estimator sample consensus algorithm based on scale invariant feature transform features is devised to compute piecewise-linear transformations to approximate the non-linear deformations of palmprints, and then, the stable regions complying with the linear transformations are decided using a block growing algorithm. Palmprint feature extraction and matching are performed over these stable regions to compute matching scores for decision. Experiments on several public palmprint databases show that the proposed models and the KPBG approach can effectively solve the deformation problem in palmprint verification and outperform the state-of-the-art methods. PMID:26390453

  4. Cowpea [Vigna unguiculata (L.) Walp].

    PubMed

    Behura, Ratikanta; Kumar, Sanjeev; Saha, Bedabrata; Panda, Manasa Kumar; Dey, Mohitosh; Sadhukhan, Ayan; Mishra, Sagarika; Alam, Shamsher; Sahoo, Debee Prasad; Sugla, Twinkle; Sahoo, Lingaraj

    2015-01-01

    Agrobacterium tumefaciens-mediated transformation is an efficient method for incorporating genes and recovering stable transgenic plants in cowpea because this method offers several advantages such as the defined integration of transgenes, potentially low copy number, and preferential integration into transcriptional active regions of the chromosome. Cotyledonary node explants of cowpea present an attractive target for T-DNA delivery followed by regeneration of shoots via axillary proliferation without involvement of a de novo regeneration pathway. In this chapter, we describe a detailed protocol for Agrobacterium-mediated transformation of the cowpea variety Pusa Komal. The seedling cotyledonary node explants are used for cocultivation with an Agrobacterium strain EHA105 harboring standard binary vector, pCAMBIA2301 or pNOV2819, and putative transformed plants are selected using aminoglycoside antibiotic or mannose as sole carbon source, respectively. The entire process includes explant infection to transgenic seed generation in greenhouse. PMID:25300846

  5. Stable coherent states

    NASA Astrophysics Data System (ADS)

    Zipfel, Antonia; Thiemann, Thomas

    2016-04-01

    We analyze the stability under time evolution of complexifier coherent states (CCS) in one-dimensional mechanical systems. A system of coherent states is called stable if it evolves into another coherent state. It turns out that a system can only possess stable CCS if the classical evolution of the variable z =e-i Lχ Cq for a given complexifier C depends only on z itself and not on its complex conjugate. This condition is very restrictive in general so that only a few systems exist that obey this condition. However, it is possible to access a wider class of models that in principle may allow for stable coherent states associated with certain regions in the phase space by introducing action-angle coordinates.

  6. SHARP transformation

    NASA Astrophysics Data System (ADS)

    Wyatt, Stephan

    2004-08-01

    The U.S. Navy"s SHAred Reconnaissance Pod (SHARP) employs the Recon/Optical, Inc. (ROI) CA-279 dual spectral band (visible/IR) digital cameras operating from an F-18E/F aircraft to perform low-to-high altitude reconnaissance missions. SHARP has proven itself combat worthy, with a rapid transition from development to operational deployment culminating in a highly reliable and effective reconnaissance capability for joint forces operating in Operation Iraqi Freedom (OIF). The U.S. Navy"s intelligence, surveillance and reconnaissance (ISR) roadmap transforms the SHARP system from being solely an independent reconnaissance sensor to a node in the growing Joint ISR network. ROI and the U.S. Navy have combined their resources to ensure the system"s transformation continues to follow the ISR road map. Pre-planned product improvements (P3I) for the CA-270 camera systems will lead the way in that transformation.

  7. Functional Characterization of a Bidirectional Plant Promoter from Cotton Leaf Curl Burewala Virus Using an Agrobacterium-Mediated Transient Assay

    PubMed Central

    Ashraf, Muhammad Aleem; Shahid, Ahmad Ali; Rao, Abdul Qayyum; Bajwa, Kamran Shehzad; Husnain, Tayyab

    2014-01-01

    The C1 promoter expressing the AC1 gene, and V1 promoter expressing the AV1 gene are located in opposite orientations in the large intergenic region of the Cotton leaf curl Burewala virus (CLCuBuV) genome. Agro-infiltration was used to transiently express putative promoter constructs in Nicotiana tabacum and Gossypium hirsutum leaves, which was monitored by a GUS reporter gene, and revealed that the bidirectional promoter of CLCuBuV transcriptionally regulates both the AC1 and AV1 genes. The CLCuBuV C1 gene promoter showed a strong, consistent transient expression of the reporter gene (GUS) in N. tabacum and G. hirsutum leaves and exhibited GUS activity two- to three-fold higher than the CaMV 35S promoter. The CLCuBuV bidirectional genepromoter is a nearly constitutive promoter that contains basic conserved elements. Many cis-regulatory elements (CREs) were also analyzed within the bidirectional plant promoters of CLCuBuV and closely related geminiviruses, which may be helpful in understanding the transcriptional regulation of both the virus and host plant. PMID:24424501

  8. Genetic transformation of cotton with a harpin-encoding gene hpaXoo confers an enhanced defense response against different pathogens through a priming mechanism

    PubMed Central

    2010-01-01

    Background The soil-borne fungal pathogen Verticillium dahliae Kleb causes Verticillium wilt in a wide range of crops including cotton (Gossypium hirsutum). To date, most upland cotton varieties are susceptible to V. dahliae and the breeding for cotton varieties with the resistance to Verticillium wilt has not been successful. Results Hpa1Xoo is a harpin protein from Xanthomonas oryzae pv. oryzae which induces the hypersensitive cell death in plants. When hpa1Xoo was transformed into the susceptible cotton line Z35 through Agrobacterium-mediated transformation, the transgenic cotton line (T-34) with an improved resistance to Verticillium dahliae was obtained. Cells of the transgenic T-34, when mixed with the conidia suspension of V. dahliae, had a higher tolerance to V. dahliae compared to cells of untransformed Z35. Cells of T-34 were more viable 12 h after mixing with V. dahliae conidia suspension. Immunocytological analysis showed that Hpa1Xoo, expressed in T-34, accumulated as clustered particles along the cell walls of T-34. In response to the infection caused by V. dahliae, the microscopic cell death and the generation of reactive oxygen intermediates were observed in leaves of T-34 and these responses were absent in leaves of Z35 inoculated with V. dahliae. Quantitative RT-PCR analysis indicated that five defense-related genes, ghAOX1, hin1, npr1, ghdhg-OMT, and hsr203J, were up-regulated in T-34 inoculated with V. dahliae. The up-regulations of these defense-relate genes were not observed or in a less extent in leaves of Z-35 after the inoculation. Conclusions Hpa1Xoo accumulates along the cell walls of the transgenic T-34, where it triggers the generation of H2O2 as an endogenous elicitor. T-34 is thus in a primed state, ready to protect the host from the pathogen. The results of this study suggest that the transformation of cotton with hpa1Xoo could be an effective approach for the development of cotton varieties with the improved resistance against soil

  9. The Stable Pairing Problem

    ERIC Educational Resources Information Center

    Greenwell, Raymond N.; Seabold, Daniel E.

    2014-01-01

    The Gale-Shapley stable marriage theorem is a fascinating piece of twentieth-century mathematics that has many practical applications--from labor markets to school admissions--yet is accessible to secondary school mathematics students. David Gale and Lloyd Shapley were both mathematicians and economists who published their work on the Stable…

  10. Stable isotope studies

    SciTech Connect

    Ishida, T.

    1992-01-01

    The research has been in four general areas: (1) correlation of isotope effects with molecular forces and molecular structures, (2) correlation of zero-point energy and its isotope effects with molecular structure and molecular forces, (3) vapor pressure isotope effects, and (4) fractionation of stable isotopes. 73 refs, 38 figs, 29 tabs.

  11. Transformation Time

    ERIC Educational Resources Information Center

    Berry, John N., III

    2007-01-01

    The program for the march by librarians on America's capital for the American Library Association (ALA) conference is predictably loaded with lobbying, legislation, and DC tours. It also abounds with professional opportunity and reflects the impact of Leslie Burger, one of the most activist ALA presidents in recent history. Her "Transformation"…

  12. Transformation & Metamorphosis

    ERIC Educational Resources Information Center

    Lott, Debra

    2009-01-01

    The sculptures of Canadian artist Brian Jungen are a great inspiration for a lesson on creating new forms. Jungen transforms found objects into unique creations without fully concealing their original form or purpose. Frank Stella's sculpture series, including "K.132,2007" made of stainless steel and spray paint, is another great example of…

  13. Transforming Schools.

    ERIC Educational Resources Information Center

    Cookson, Peter W., Jr., Ed.; Schneider, Barbara, Ed.

    The authors in this book address the issues that relate to the crisis in American education and review some of the proposed solutions. To transform education, schools must be examined as social systems that are interrelated with families, communities, and the world of work. Following the introduction, section 1, "Conditions for Educational…

  14. Stable isotopes in mineralogy

    USGS Publications Warehouse

    O'Neil, J.R.

    1977-01-01

    Stable isotope fractionations between minerals are functions of the fundamental vibrational frequencies of the minerals and therefore bear on several topics of mineralogical interest. Isotopic compositions of the elements H, C, O, Si, and S can now be determined routinely in almost any mineral. A summary has been made of both published and new results of laboratory investigations, analyses of natural materials, and theoretical considerations which bear on the importance of temperature, pressure, chemical composition and crystal structure to the isotopic properties of minerals. It is shown that stable isotope studies can sometimes provide evidence for elucidating details of crystal structure and can be a powerful tool for use in tracing the reaction paths of mineralogical reactions. ?? 1977 Springer-Verlag.

  15. Stable local oscillator module.

    SciTech Connect

    Brocato, Robert Wesley

    2007-11-01

    This report gives a description of the development of a Stable Local Oscillator (StaLO) multi-chip module (MCM). It is a follow-on report to SAND2006-6414, Stable Local Oscillator Microcircuit. The StaLO accepts a 100MHz input signal and produces output signals at 1.2, 3.3, and 3.6 GHz. The circuit is built as a multi-chip module (MCM), since it makes use of integrated circuit technologies in silicon and lithium niobate as well as discrete passive components. This report describes the development of an MCM-based version of the complete StaLO, fabricated on an alumina thick film hybrid substrate.

  16. Handbook of stable strontium

    SciTech Connect

    Skoryna, S.C.

    1981-01-01

    This book presents information on the following topics: chemistry of strontium; biogeochemistry of strontium; uptake of stable strontium by plants and effects on plant growth; divalent cation-dependent deposits in paramecium; effects of strontium ion on the hydrolysis of ATP; stronium ions and membranes - screening versus binding at charged surfaces; mitochondrial granules in the liver of rats kept on stable strontium supplementation; divalent cations and regulation of cyclic nucleotides in nervous systems; strontium as the substitute for calcium in the excitation-contraction coupling of crayfish muscle fibers; hemodynamic effects of strontium in the dog; some mechanical characteristics of strontium-mediated contractions in heart muscle; effects of calcium, magnesium, and strontium on drug-receptor interactions; strontium and histamine secretion; and effects of strontium in human dental enamel.

  17. Stable Charged Cosmic Strings

    SciTech Connect

    Weigel, H.; Quandt, M.; Graham, N.

    2011-03-11

    We study the quantum stabilization of a cosmic string by a heavy fermion doublet in a reduced version of the standard model. We show that charged strings, obtained by populating fermionic bound state levels, become stable if the electroweak bosons are coupled to a fermion that is less than twice as heavy as the top quark. This result suggests that extraordinarily large fermion masses or unrealistic couplings are not required to bind a cosmic string in the standard model. Numerically we find the most favorable string profile to be a simple trough in the Higgs vacuum expectation value of radius {approx_equal}10{sup -18} m. The vacuum remains stable in our model, because neutral strings are not energetically favored.

  18. Stable charged cosmic strings.

    PubMed

    Weigel, H; Quandt, M; Graham, N

    2011-03-11

    We study the quantum stabilization of a cosmic string by a heavy fermion doublet in a reduced version of the standard model. We show that charged strings, obtained by populating fermionic bound state levels, become stable if the electroweak bosons are coupled to a fermion that is less than twice as heavy as the top quark. This result suggests that extraordinarily large fermion masses or unrealistic couplings are not required to bind a cosmic string in the standard model. Numerically we find the most favorable string profile to be a simple trough in the Higgs vacuum expectation value of radius ≈10(-18)  m. The vacuum remains stable in our model, because neutral strings are not energetically favored. PMID:21469786

  19. RF transformer

    DOEpatents

    Smith, James L.; Helenberg, Harold W.; Kilsdonk, Dennis J.

    1979-01-01

    There is provided an improved RF transformer having a single-turn secondary of cylindrical shape and a coiled encapsulated primary contained within the secondary. The coil is tapered so that the narrowest separation between the primary and the secondary is at one end of the coil. The encapsulated primary is removable from the secondary so that a variety of different capacity primaries can be utilized with one secondary.

  20. Thermally stable laminating resins

    NASA Technical Reports Server (NTRS)

    Jones, R. J.; Vaughan, R. W.; Burns, E. A.

    1972-01-01

    Improved thermally stable laminating resins were developed based on the addition-type pyrolytic polymerization. Detailed monomer and polymer synthesis and characterization studies identified formulations which facilitate press molding processing and autoclave fabrication of glass and graphite fiber reinforced composites. A specific resin formulation, termed P10P was utilized to prepare a Courtaulds HMS reinforced simulated airfoil demonstration part by an autoclave molding process.

  1. Forensic Stable Isotope Biogeochemistry

    NASA Astrophysics Data System (ADS)

    Cerling, Thure E.; Barnette, Janet E.; Bowen, Gabriel J.; Chesson, Lesley A.; Ehleringer, James R.; Remien, Christopher H.; Shea, Patrick; Tipple, Brett J.; West, Jason B.

    2016-06-01

    Stable isotopes are being used for forensic science studies, with applications to both natural and manufactured products. In this review we discuss how scientific evidence can be used in the legal context and where the scientific progress of hypothesis revisions can be in tension with the legal expectations of widely used methods for measurements. Although this review is written in the context of US law, many of the considerations of scientific reproducibility and acceptance of relevant scientific data span other legal systems that might apply different legal principles and therefore reach different conclusions. Stable isotopes are used in legal situations for comparing samples for authenticity or evidentiary considerations, in understanding trade patterns of illegal materials, and in understanding the origins of unknown decedents. Isotope evidence is particularly useful when considered in the broad framework of physiochemical processes and in recognizing regional to global patterns found in many materials, including foods and food products, drugs, and humans. Stable isotopes considered in the larger spatial context add an important dimension to forensic science.

  2. Hamlet's Transformation.

    NASA Astrophysics Data System (ADS)

    Usher, P. D.

    1997-12-01

    William Shakespeare's Hamlet has much evidence to suggest that the Bard was aware of the cosmological models of his time, specifically the geocentric bounded Ptolemaic and Tychonic models, and the infinite Diggesian. Moreover, Shakespeare describes how the Ptolemaic model is to be transformed to the Diggesian. Hamlet's "transformation" is the reason that Claudius, who personifies the Ptolemaic model, summons Rosencrantz and Guildenstern, who personify the Tychonic. Pantometria, written by Leonard Digges and his son Thomas in 1571, contains the first technical use of the word "transformation." At age thirty, Thomas Digges went on to propose his Perfit Description, as alluded to in Act Five where Hamlet's age is given as thirty. In Act Five as well, the words "bore" and "arms" refer to Thomas' vocation as muster-master and his scientific interest in ballistics. England's leading astronomer was also the father of the poet whose encomium introduced the First Folio of 1623. His oldest child Dudley became a member of the Virginia Company and facilitated the writing of The Tempest. Taken as a whole, such manifold connections to Thomas Digges support Hotson's contention that Shakespeare knew the Digges family. Rosencrantz and Guildenstern in Hamlet bear Danish names because they personify the Danish model, while the king's name is latinized like that of Claudius Ptolemaeus. The reason Shakespeare anglicized "Amleth" to "Hamlet" was because he saw a parallel between Book Three of Saxo Grammaticus and the eventual triumph of the Diggesian model. But Shakespeare eschewed Book Four, creating this particular ending from an infinity of other possibilities because it "suited his purpose," viz. to celebrate the concept of a boundless universe of stars like the Sun.

  3. TRANSFORMER APPARATUS

    DOEpatents

    Wolfgang, F.; Nicol, J.

    1962-11-01

    Transformer apparatus is designed for measuring the amount of a paramagnetic substance dissolved or suspended in a diamagnetic liquid. The apparatus consists of a cluster of tubes, some of which are closed and have sealed within the diamagnetic substance without any of the paramagnetic material. The remaining tubes are open to flow of the mix- ture. Primary and secondary conductors are wrapped around the tubes in such a way as to cancel noise components and also to produce a differential signal on the secondaries based upon variations of the content of the paramagnetic material. (AEC)

  4. Rotary Transformer

    NASA Technical Reports Server (NTRS)

    McLyman, Colonel Wm. T.

    1996-01-01

    None given. From first Par: Many spacecraft (S/C) and surface rovers require the transfer of signals and power across rotating interfaces. Science instruments, antennas and solar arrays are elements needing rotary power transfer for certain (S/C) configurations. Delivery of signal and power has mainly been done by using the simplest means, the slip ring approach. This approach, although simple, leaves debris generating noise over a period of time...The rotary transformer is a good alternative to slip rings for signal and power transfer.

  5. Corn transformed

    SciTech Connect

    Moffat, A.S.

    1990-08-10

    Researchers have produced fertile corn transformed with a foreign gene that makes the plants resistant to the herbicide bialaphos. This achievement, is the first report of fertile transgenic corn in the reviewed literature, and it is the capstone of almost a decade's efforts to genetically engineer this country's most important crop. The only other major crop to be so manipulated is rice. The ability produce transgenic corn gives biologists a valuable tool to probe the whys and hows of gene expression and regulation. It may also give plant breeders a way to develop new corn varieties with a speed and predictability that would be impossible with classical breeding techniques.

  6. Transformational leadership.

    PubMed

    Marlow, D L

    1996-01-01

    In these uncertain times in the healthcare industry, administrators are asked to do more with less time and resources. Because of the extended roles they are playing in today's organizations, radiology administrators are looked upon as agents of change. What leadership skills do they need in this turbulent and uncertain healthcare environment? What are the trait's of tomorrow's leaders? The transformational leader is the one who will guide us through this changing healthcare environment. Several behavioral patterns emerge as important traits for tomorrow's leaders to have-individual consideration, intellectual stimulation and charisma. Tomorrow's leader must view each person as an individual, showing genuine concern and belief in each person's ability to perform. Transformational leaders stimulate others by encouraging them to be curious and try new ideas. The final characteristic, charisma, is the ability to inspire others. Luckily, leaders are made, not born: today's leaders can learn to be responsive, to draw out new ideas from employees, and to communicate self-esteem, energy and enthusiasm. PMID:10163135

  7. Stable predictive control horizons

    NASA Astrophysics Data System (ADS)

    Estrada, Raúl; Favela, Antonio; Raimondi, Angelo; Nevado, Antonio; Requena, Ricardo; Beltrán-Carbajal, Francisco

    2012-04-01

    The stability theory of predictive and adaptive predictive control for processes of linear and stable nature is based on the hypothesis of a physically realisable driving desired trajectory (DDT). The formal theoretical verification of this hypothesis is trivial for processes with a stable inverse, but it is not for processes with an unstable inverse. The extended strategy of predictive control was developed with the purpose of overcoming methodologically this stability problem and it has delivered excellent performance and stability in its industrial applications given a suitable choice of the prediction horizon. From a theoretical point of view, the existence of a prediction horizon capable of ensuring stability for processes with an unstable inverse was proven in the literature. However, no analytical solution has been found for the determination of the prediction horizon values which guarantee stability, in spite of the theoretical and practical interest of this matter. This article presents a new method able to determine the set of prediction horizon values which ensure stability under the extended predictive control strategy formulation and a particular performance criterion for the design of the DDT generically used in many industrial applications. The practical application of this method is illustrated by means of simulation examples.

  8. A piezoelectric transformer

    NASA Technical Reports Server (NTRS)

    Won, C. C.

    1993-01-01

    This work describes a modeling and design method whereby a piezoelectric system is formulated by two sets of second-order equations, one for the mechanical system, and the other for the electrical system, coupled through the piezoelectric effect. The solution to this electromechanical coupled system gives a physical interpretation of the piezoelectric effect as a piezoelectric transformer that is a part of the piezoelectric system, which transfers the applied mechanical force into a force-controlled current source, and short circuit mechanical compliance into capacitance. It also transfers the voltage source into a voltage-controlled relative velocity input, and free motional capacitance into mechanical compliance. The formulation and interpretation simplify the modeling of smart structures and lead to physical insight that aids the designer. Due to its physical realization, the smart structural system can be unconditional stable and effectively control responses. This new concept has been demonstrated in three numerical examples for a simple piezoelectric system.

  9. Marginally Stable Nuclear Burning

    NASA Technical Reports Server (NTRS)

    Strohmayer, Tod E.; Altamirano, D.

    2012-01-01

    Thermonuclear X-ray bursts result from unstable nuclear burning of the material accreted on neutron stars in some low mass X-ray binaries (LMXBs). Theory predicts that close to the boundary of stability oscillatory burning can occur. This marginally stable regime has so far been identified in only a small number of sources. We present Rossi X-ray Timing Explorer (RXTE) observations of the bursting, high- inclination LMXB 4U 1323-619 that reveal for the first time in this source the signature of marginally stable burning. The source was observed during two successive RXTE orbits for approximately 5 ksec beginning at 10:14:01 UTC on March 28, 2011. Significant mHz quasi- periodic oscillations (QPO) at a frequency of 8.1 mHz are detected for approximately 1600 s from the beginning of the observation until the occurrence of a thermonuclear X-ray burst at 10:42:22 UTC. The mHz oscillations are not detected following the X-ray burst. The average fractional rms amplitude of the mHz QPOs is 6.4% (3 - 20 keV), and the amplitude increases to about 8% below 10 keV.This phenomenology is strikingly similar to that seen in the LMXB 4U 1636-53. Indeed, the frequency of the mHz QPOs in 4U 1323-619 prior to the X-ray burst is very similar to the transition frequency between mHz QPO and bursts found in 4U 1636-53 by Altamirano et al. (2008). These results strongly suggest that the observed QPOs in 4U 1323-619 are, like those in 4U 1636-53, due to marginally stable nuclear burning. We also explore the dependence of the energy spectrum on the oscillation phase, and we place the present observations within the context of the spectral evolution of the accretion-powered flux from the source.

  10. Herbicide Transformation

    PubMed Central

    Lanzilotta, R. P.; Pramer, David

    1970-01-01

    A strain of Fusarium solani isolated from soil by enrichment techniques used propanil (3′, 4′-dichloropropionanilide) as a sole source of organic carbon and energy for growth in pure culture. The primary product of the transformation of propanil by F. solani was isolated and identified as 3,4-dichloroaniline (DCA). This compound accumulated in the medium to a level (80 μg/ml) which stopped further herbicide utilization. Herbicide utilization by F. solani was influenced by various environmental and nutritional factors. It was more sensitive to acid than alkaline pH. Added glucose and yeast extract increased the rate of propanil decomposition, and the reduced aeration retarded growth of the fungus and herbicide utilization. The growth of F. solani on propionate was inhibited by added DCA. Images PMID:5437305

  11. Stable local oscillator microcircuit.

    SciTech Connect

    Brocato, Robert Wesley

    2006-10-01

    This report gives a description of the development of a Stable Local Oscillator (StaLO) Microcircuit. The StaLO accepts a 100MHz input signal and produces output signals at 1.2, 3.3, and 3.6 GHz. The circuit is built as a multi-chip module (MCM), since it makes use of integrated circuit technologies in silicon and lithium niobate as well as discrete passive components. The StaLO uses a comb generator followed by surface acoustic wave (SAW) filters. The comb generator creates a set of harmonic components of the 100MHz input signal. The SAW filters are narrow bandpass filters that are used to select the desired component and reject all others. The resulting circuit has very low sideband power levels and low phase noise (both less than -40dBc) that is limited primarily by the phase noise level of the input signal.

  12. Stable magnetic universes revisited

    NASA Astrophysics Data System (ADS)

    Tahamtan, T.; Halilsoy, M.

    2013-01-01

    Recent discovery of magnetars ( B˜1015 G) motivates us to consider magnetic universes in general relativity a new. A regular class of static, cylindrically symmetric pure magnetic field metrics is rederived in a different metric ansatz in all dimensions. Radial, time dependent perturbations show that for dimensions d>3 such spacetimes are stable at both near r≈0 and large radius r→∞. For d=3, however, simultaneous stability requirement at both, near and far radial distances can not be reconciled for time-dependent perturbations. We argue that this distinct property may be the cause for the absence of pure magnetic black holes in d=3. Restricted, numerical geodesics for neutral particles reveal a gravitational confinement around the center in the polar plane. Charged, time-like geodesics for d=4 are shown numerically to remain confined as well.

  13. Economic Impact of Stable Flies

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A dynamic model was created to estimate the economic impact of stable flies on livestock production. Based upon a nationwide average of 10 stable flies per animal for 3 months per year, the model estimates the impact of stable flies to be $543 million to the dairy industry, $1.34 billion to pasture ...

  14. Bi-stable optical actuator

    DOEpatents

    Holdener, Fred R.; Boyd, Robert D.

    2000-01-01

    The present invention is a bi-stable optical actuator device that is depowered in both stable positions. A bearing is used to transfer motion and smoothly transition from one state to another. The optical actuator device may be maintained in a stable position either by gravity or a restraining device.

  15. Stable umbral chromospheric structures

    NASA Astrophysics Data System (ADS)

    Henriques, V. M. J.; Scullion, E.; Mathioudakis, M.; Kiselman, D.; Gallagher, P. T.; Keenan, F. P.

    2015-02-01

    Aims: We seek to understand the morphology of the chromosphere in sunspot umbra. We investigate if the horizontal structures observed in the spectral core of the Ca II H line are ephemeral visuals caused by the shock dynamics of more stable structures, and examine their relationship with observables in the H-alpha line. Methods: Filtergrams in the core of the Ca II H and H-alpha lines as observed with the Swedish 1-m Solar Telescope are employed. We utilise a technique that creates composite images and tracks the flash propagation horizontally. Results: We find 0.̋15 wide horizontal structures, in all of the three target sunspots, for every flash where the seeing is moderate to good. Discrete dark structures are identified that are stable for at least two umbral flashes, as well as systems of structures that live for up to 24 min. We find cases of extremely extended structures with similar stability, with one such structure showing an extent of 5''. Some of these structures have a correspondence in H-alpha, but we were unable to find a one-to-one correspondence for every occurrence. If the dark streaks are formed at the same heights as umbral flashes, there are systems of structures with strong departures from the vertical for all three analysed sunspots. Conclusions: Long-lived Ca II H filamentary horizontal structures are a common and likely ever-present feature in the umbra of sunspots. If the magnetic field in the chromosphere of the umbra is indeed aligned with the structures, then the present theoretical understanding of the typical umbra needs to be revisited. Movies associated to Figs. 3 and 4 are available in electronic form at http://www.aanda.org

  16. Stable superstring relics

    SciTech Connect

    Chang, S.; Coriano, C.; Faraggi, A.E. |

    1996-05-15

    The authors investigate the cosmological constraints on exotic stable matter states which arise in realistic free fermionic superstring models. These states appear in the superstring models due to a ``Wilson-line`` breaking of the unifying non-Abelian gauge symmetry. In the models that they consider the unifying SO(10) gauge symmetry is broken at the string level to SO(6) x SO(4), SU(5) x U(1) or SU(3) x SU(2) x U(1). The exotic matter states are classified according to the patterns of the SO(10) symmetry breaking. In SO(6) x XO(4) and SU(5) x U(1) type models one obtains fractionally charged states with Q{sub e.m.} = {+-}1/2. In SU(3) x SU(2) x U(1) type models one also obtains states with the regular charges under the Standard Model gauge group but with ``fractional`` charges under the U(1){sub z{prime}} symmetry. These states include down-like color triplets and electroweak doublets, as well as states which are Standard Model singlets. By analyzing the renormalizable and nonrenormalizable terms of the superpotential in a specific superstring model, the authors show that these exotic states can be stable. They investigate the cosmological constraints on the masses and relic density of the exotic states. They propose that, while the abundance and the masses of the fractionally charged states are highly constrained, the Standard Model-like states, and in particular the Standard Model singlet, are good dark matter candidates.

  17. Stable face representations

    PubMed Central

    Jenkins, Rob; Burton, A. Mike

    2011-01-01

    Photographs are often used to establish the identity of an individual or to verify that they are who they claim to be. Yet, recent research shows that it is surprisingly difficult to match a photo to a face. Neither humans nor machines can perform this task reliably. Although human perceivers are good at matching familiar faces, performance with unfamiliar faces is strikingly poor. The situation is no better for automatic face recognition systems. In practical settings, automatic systems have been consistently disappointing. In this review, we suggest that failure to distinguish between familiar and unfamiliar face processing has led to unrealistic expectations about face identification in applied settings. We also argue that a photograph is not necessarily a reliable indicator of facial appearance, and develop our proposal that summary statistics can provide more stable face representations. In particular, we show that image averaging stabilizes facial appearance by diluting aspects of the image that vary between snapshots of the same person. We review evidence that the resulting images can outperform photographs in both behavioural experiments and computer simulations, and outline promising directions for future research. PMID:21536553

  18. Unusually stable liquid foams.

    PubMed

    Rio, Emmanuelle; Drenckhan, Wiebke; Salonen, Anniina; Langevin, Dominique

    2014-03-01

    Obtaining stable liquid foams is an important issue in view of their numerous applications. In some of these, the liquid foam in itself is of interest, in others, the liquid foam acts as a precursor for the generation of solid foam. In this short review, we will make a survey of the existing results in the area. This will include foams stabilised by surfactants, proteins and particles. The origin of the stability is related to the slowing down of coarsening, drainage or coalescence, and eventually to their arrest. The three effects are frequently coupled and in many cases, they act simultaneously and enhance one another. Drainage can be arrested if the liquid of the foam either gels or solidifies. Coalescence is slowed down by gelified foam films, and it can be arrested if the films become very thick and/or rigid. These mechanisms are thus qualitatively easy to identify, but they are less easy to model in order to obtain quantitative predictions. The slowing down of coarsening requests either very thick or small films, and its arrest was observed in cases where the surface compression modulus was large. The detail of the mechanisms at play remains unclear. PMID:24342735

  19. Construction of the first phage library of chickpea blight pathogen Ascochyta rabiei.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During studies on pathogenic determinants of Ascochyta rabiei, the causal agent of chickpea blight, we developed a library of insertional mutants of A. rabiei using Agrobacterium - mediated transformation. Non-pathogenic mutants were identified after screening more than 1000 transformants using path...

  20. CRISPR/Cas-Mediated Site-Specific Mutagenesis in Arabidopsis thaliana Using Cas9 Nucleases and Paired Nickases.

    PubMed

    Schiml, Simon; Fauser, Friedrich; Puchta, Holger

    2016-01-01

    The CRISPR/Cas system has recently become the most important tool for genome engineering due to its simple architecture that allows for rapidly changing the target sequence and its applicability to organisms throughout all kingdoms of life. The need for an easy-to-use and reliable nuclease is especially high in plant research, as precise genome modifications are almost impossible to achieve by Agrobacterium-mediated transformation and the regeneration of plants from protoplast cultures is very labor intensive. Here, we describe the application of the Cas9 nuclease to Arabidopsis thaliana for the induction of heritable targeted mutations, which may also be used for other plant species. To cover the concern for off-target activity, we also describe the generation of stable mutants using paired Cas9 nickases. PMID:27557689

  1. Stable isotope laser spectroscopy

    NASA Technical Reports Server (NTRS)

    Becker, J. F.; Yaldaei, Ramil; Mckay, Christopher P.

    1989-01-01

    Recent advances in semiconductor laser technology have produced a reliable lightweight device ideally suited for a spacecraft high resolution molecular spectrometer. Lead-salt tunable diode lasers (TDL) emit in several spectral modes, each with a very narrow linewidth of -0.0003/cm. This spectral resolution is much narrower than typical Doppler broadened molecular linewidths in the mid-IR range. Thus it is possible to detect individual rotational lines within the vibrational band and measure their intensity, which can be used to determine gas concentration. The narrow spectral lines of any impurity gas tend to lie between the narrow lines of the gas of interest. This represents a major advantage over the accepted gas chromatograph mass spectrometer (GCMS) technique for measuring gas concentrations and isotope ratios. The careful and extensive gas purification procedures required to remove impurities for reliable GCMS measurements will not be required for an IR laser gas analysis. The infrared laser gas analysis technique is being developed to measure stable isotopic ratios of gases such as CO2, CH4, N2O, and NH3. This will eventually lead to development of instruments capable of in situ istopic measurements on planets such as Mars. The carbon (C-12, C-13) isotope ratio is indicative of the type of carbon fixation mechanisms (e.g., photosynthesis, respiration) in operation on a planet, while the nitrogen (N-14, N-15) isotope ratio can probably be used to date nitrogen-bearing Martian samples. The absorbance ratio of two adjacent lines of CO2 in the 2300/cm (4.3 micron) region of the spectrum was measured. The precision of the measurement is presently better than 1 percent and significant improvement is anticipated as rapid sweep-integration techniques and computer controlled data acquistion capabilities are incorporated.

  2. Stable isotope laser spectroscopy

    NASA Astrophysics Data System (ADS)

    Becker, J. F.; Yaldaei, Ramil; McKay, Christopher P.

    1989-03-01

    Recent advances in semiconductor laser technology have produced a reliable lightweight device ideally suited for a spacecraft high resolution molecular spectrometer. Lead-salt tunable diode lasers (TDL) emit in several spectral modes, each with a very narrow linewidth of -0.0003/cm. This spectral resolution is much narrower than typical Doppler broadened molecular linewidths in the mid-IR range. Thus it is possible to detect individual rotational lines within the vibrational band and measure their intensity, which can be used to determine gas concentration. The narrow spectral lines of any impurity gas tend to lie between the narrow lines of the gas of interest. This represents a major advantage over the accepted gas chromatograph mass spectrometer (GCMS) technique for measuring gas concentrations and isotope ratios. The careful and extensive gas purification procedures required to remove impurities for reliable GCMS measurements will not be required for an IR laser gas analysis. The infrared laser gas analysis technique is being developed to measure stable isotopic ratios of gases such as CO2, CH4, N2O, and NH3. This will eventually lead to development of instruments capable of in situ istopic measurements on planets such as Mars. The carbon (C-12, C-13) isotope ratio is indicative of the type of carbon fixation mechanisms (e.g., photosynthesis, respiration) in operation on a planet, while the nitrogen (N-14, N-15) isotope ratio can probably be used to date nitrogen-bearing Martian samples. The absorbance ratio of two adjacent lines of CO2 in the 2300/cm (4.3 micron) region of the spectrum was measured. The precision of the measurement is presently better than 1 percent and significant improvement is anticipated as rapid sweep-integration techniques and computer controlled data acquistion capabilities are incorporated.

  3. Stable electroosmotically driven actuators

    NASA Astrophysics Data System (ADS)

    Sritharan, Deepa; Motsebo, Mylene; Tumbic, Julia; Smela, Elisabeth

    2013-04-01

    We have previously presented "nastic" actuators based on electroosmotic (EO) pumping of fluid in microchannels using high electric fields for potential application in soft robotics. In this work we address two challenges facing this technology: applying EO to meso-scale devices and the stability of the pumping fluid. The hydraulic pressure achieved by EO increases with as 1/d2, where d is the depth of the microchannel, but the flow rate (which determines the stroke and the speed) is proportional to nd, where n is the number of channels. Therefore to get high force and high stroke the device requires a large number of narrow channels, which is not readily achievable using standard microfabrication techniques. Furthermore, for soft robotics the structure must be soft. In this work we present a method of fabricating a three-dimensional porous elastomer to serve as the array of channels based on a sacrificial sugar scaffold. We demonstrate the concept by fabricating small pumps. The flexible devices were made from polydimethylsiloxane (PDMS) and comprise the 3D porous elastomer flanked on either side by reservoirs containing electrodes. The second issue addressed here involves the pumping fluid. Typically, water is used for EO, but water undergoes electrolysis even at low voltages. Since EO takes place at kV, these systems must be open to release the gases. We have recently reported that propylene carbonate (PC) is pumped at a comparable rate as water and is also stable for over 30 min at 8 kV. Here we show that PC is, however, degraded by moisture, so future EO systems must prevent water from reaching the PC.

  4. Optimal conditions for transformation of Azotobacter vinelandii.

    PubMed

    Page, W J; von Tigerstrom, M

    1979-09-01

    Optimal transformation of Azotobacter vinelandii OP required a 20-min incubation of the competent cells with deoxyribonucleic acid at 30 degrees C in buffer (pH 6.0 to 8.0) containing 8 mM magnesium sulfate. Nitrogen-fixing transformants of nitrogen fixation-deficient recipients could be plated immediately on selective medium, but transformants acquiring rifampin and streptomycin resistance required preincubation in nonselective medium. The three phenotypes achieved an approximately equal and stable frequency after 17 h (six generations) of growth in nonselective medium. PMID:479104

  5. Discrete radon transform with shift of coordinate

    SciTech Connect

    Wei, X.; Wu, L.

    1994-12-31

    This paper describes the Radon transform with shift of coordinates. The appropriate shift value of spatial coordinate gives less alias in the data reconstruction and the appropriate shift value of frequency coordinate makes the reconstruction stable. The method is suitable to signal processing of seismogram, an example of wave field separation to practical VSP data is shown in the paper.

  6. Nuclear transformation of Volvox carteri.

    PubMed Central

    Schiedlmeier, B; Schmitt, R; Müller, W; Kirk, M M; Gruber, H; Mages, W; Kirk, D L

    1994-01-01

    Stable nuclear transformation of Volvox carteri was achieved using the cloned V. carteri nitA+ gene (which encodes nitrate reductase) to complement a nitA- mutation. Following bombardment of mutant cells with plasmid-coated gold particles, putative transformants able to utilize nitrate as a nitrogen source were recovered with an efficiency of approximately 2.5 x 10(5). DNA analysis indicated that the plasmid integrated into the genome, often in multiple copies, at sites other than the nitA locus. Cotransformants were recovered with a frequency of 40-80% when cells were cobombarded with a selected and an unselected marker. Thus, V. carteri becomes one of the simplest multicellular organisms that is accessible to detailed molecular studies of genes regulating cellular differentiation and morphogenesis. Images PMID:8197189

  7. Stable isotope investigations of chlorinated aliphatic hydrocarbons.

    SciTech Connect

    Abrajano, T.; Heraty, L. J.; Holt, B. D.; Huang, L.; Sturchio, N. C.

    1999-06-01

    Stable isotope ratio measurements for carbon (C) and chlorine (Cl) can be used to elucidate the processes affecting transformation and transportation of chlorinated aliphatic hydrocarbons (CAHs) in the environment. Methods recently developed in our laboratory for isotopic analysis of CAHs have been applied to laboratory measurements of the kinetic isotope effects associated with aerobic degradation of dichloromethane (DCM) and with both anaerobic and aerobic cometabolic degradation of trichlomethene (TCE) in batch and column microbial cultures. These experimental determinations of fractionation factors are crucial for understanding the behavior of CAHs in complex natural systems, where the extent of biotransformation can be masked by dispersion and volatilization. We have also performed laboratory investigations of kinetic isotope effects accompanying evaporation of CAHs, as well as field investigations of natural attenuation and in situ remediation of CAHs in a number of contaminated shallow aquifers at sites operated by the federal government and the private sector.

  8. Stable Multibubble Sonoluminescence Bubble Patterns

    SciTech Connect

    Posakony, Gerald J.; Greenwood, Lawrence R.; Ahmed, Salahuddin

    2006-06-30

    Multibubble standing wave patterns can be generated from a flat piezoceramic transducer element propagating into water. By adding a second transducer positioned at 90 degrees from the transducer generating the standing wave, a 3-dimensional volume of stable single bubbles can be established. Further, the addition of the second transducer stabilizes the bubble pattern so that individual bubbles may be studied. The size of the bubbles and the separation of the standing waves depend on the frequency of operation. Two transducers, operating at frequencies above 500 kHz, provided the most graphic results for the configuration used in this study. At these frequencies stable bubbles exhibit a bright sonoluminescence pattern. Whereas stable SBSL is well-known, stable MBSL has not been previously reported. This paper includes discussions of the acoustic responses, standing wave patterns, and pictorial results of the separation of individual bubble of sonoluminescence in a multibubble sonoluminescence environment.

  9. Earthquakes in Stable Continental Crust.

    ERIC Educational Resources Information Center

    Johnston, Arch C.; Kanter, Lisa R.

    1990-01-01

    Discussed are some of the reasons for earthquakes which occur in stable crust away from familiar zones at the ends of tectonic plates. Crust stability and the reactivation of old faults are described using examples from India and Australia. (CW)

  10. Shelf-Stable Food Safety

    MedlinePlus

    ... process of packing meat and poultry in glass bottles, corking them, and submerging them in boiling water. ... fsis.usda.gov. [ Top of Page ] Are any egg products shelf stable? Pasteurized, dried egg products can ...

  11. Hadamard Factorization of Stable Polynomials

    NASA Astrophysics Data System (ADS)

    Loredo-Villalobos, Carlos Arturo; Aguirre-Hernández, Baltazar

    2011-11-01

    The stable (Hurwitz) polynomials are important in the study of differential equations systems and control theory (see [7] and [19]). A property of these polynomials is related to Hadamard product. Consider two polynomials p,q ∈ R[x]:p(x) = anxn+an-1xn-1+...+a1x+a0q(x) = bmx m+bm-1xm-1+...+b1x+b0the Hadamard product (p × q) is defined as (p×q)(x) = akbkxk+ak-1bk-1xk-1+...+a1b1x+a0b0where k = min(m,n). Some results (see [16]) shows that if p,q ∈R[x] are stable polynomials then (p×q) is stable, also, i.e. the Hadamard product is closed; however, the reciprocal is not always true, that is, not all stable polynomial has a factorization into two stable polynomials the same degree n, if n> 4 (see [15]).In this work we will give some conditions to Hadamard factorization existence for stable polynomials.

  12. Quantitative microbial ecology through stable isotope probing.

    PubMed

    Hungate, Bruce A; Mau, Rebecca L; Schwartz, Egbert; Caporaso, J Gregory; Dijkstra, Paul; van Gestel, Natasja; Koch, Benjamin J; Liu, Cindy M; McHugh, Theresa A; Marks, Jane C; Morrissey, Ember M; Price, Lance B

    2015-11-01

    Bacteria grow and transform elements at different rates, and as yet, quantifying this variation in the environment is difficult. Determining isotope enrichment with fine taxonomic resolution after exposure to isotope tracers could help, but there are few suitable techniques. We propose a modification to stable isotope probing (SIP) that enables the isotopic composition of DNA from individual bacterial taxa after exposure to isotope tracers to be determined. In our modification, after isopycnic centrifugation, DNA is collected in multiple density fractions, and each fraction is sequenced separately. Taxon-specific density curves are produced for labeled and nonlabeled treatments, from which the shift in density for each individual taxon in response to isotope labeling is calculated. Expressing each taxon's density shift relative to that taxon's density measured without isotope enrichment accounts for the influence of nucleic acid composition on density and isolates the influence of isotope tracer assimilation. The shift in density translates quantitatively to isotopic enrichment. Because this revision to SIP allows quantitative measurements of isotope enrichment, we propose to call it quantitative stable isotope probing (qSIP). We demonstrated qSIP using soil incubations, in which soil bacteria exhibited strong taxonomic variations in (18)O and (13)C composition after exposure to [(18)O]water or [(13)C]glucose. The addition of glucose increased the assimilation of (18)O into DNA from [(18)O]water. However, the increase in (18)O assimilation was greater than expected based on utilization of glucose-derived carbon alone, because the addition of glucose indirectly stimulated bacteria to utilize other substrates for growth. This example illustrates the benefit of a quantitative approach to stable isotope probing. PMID:26296731

  13. Virus induced gene silencing in Lolium temulentum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lolium temulentum L. is valuable as a model species for studying abiotic stress in closely related forage and turf grasses, many of which are polyploid outcrossing species. As with most monocot species, Agrobacterium-mediated transformation of L. temulentum is still challenging, time consuming and n...

  14. Transgene expression in pear (Pyrus communis L.) driven by a phloem-specific promoter

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A gene expression cassette carrying ß-glucuronidase (uidA) reporter gene under the control of the promoter of the Arabidopsis sucrose-H+ symporter gene (AtSUC2) was introduced to pear plants via an Agrobacterium-mediated leaf-explant transformation procedure. Transgenic shoots were regenerated from...

  15. Mixture Screening and Mixture-Amount Designs to Determine Plant Growth Regulator Effects on Shoot Regeneration from Grapefruit (Citrus paradisi Macf.) Epicotyls

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to improve shoot regeneration from grapefruit epicotyl explants since some important in vitro applications in citrus, such as Agrobacterium-mediated transformation, commonly use epicotyl segments from in vitro seedlings; adequate adventitious shoot production is thus...

  16. Metal stable isotope signatures as tracers in environmental geochemistry.

    PubMed

    Wiederhold, Jan G

    2015-03-01

    The biogeochemical cycling of metals in natural systems is often accompanied by stable isotope fractionation which can now be measured due to recent analytical advances. In consequence, a new research field has emerged over the last two decades, complementing the traditional stable isotope systems (H, C, O, N, S) with many more elements across the periodic table (Li, B, Mg, Si, Cl, Ca, Ti, V, Cr, Fe, Ni, Cu, Zn, Ge, Se, Br, Sr, Mo, Ag, Cd, Sn, Sb, Te, Ba, W, Pt, Hg, Tl, U) which are being explored and potentially applicable as novel geochemical tracers. This review presents the application of metal stable isotopes as source and process tracers in environmental studies, in particular by using mixing and Rayleigh model approaches. The most important concepts of mass-dependent and mass-independent metal stable isotope fractionation are introduced, and the extent of natural isotopic variations for different elements is compared. A particular focus lies on a discussion of processes (redox transformations, complexation, sorption, precipitation, dissolution, evaporation, diffusion, biological cycling) which are able to induce metal stable isotope fractionation in environmental systems. Additionally, the usefulness and limitations of metal stable isotope signatures as tracers in environmental geochemistry are discussed and future perspectives presented. PMID:25640608

  17. Transforming the Way We Teach Function Transformations

    ERIC Educational Resources Information Center

    Faulkenberry, Eileen Durand; Faulkenberry, Thomas J.

    2010-01-01

    In this article, the authors discuss "function," a well-defined rule that relates inputs to outputs. They have found that by using the input-output definition of "function," they can examine transformations of functions simply by looking at changes to input or output and the respective changes to the graph. Applying transformations to the input…

  18. Symplectic wavelet transformation.

    PubMed

    Fan, Hong-Yi; Lu, Hai-Liang

    2006-12-01

    Usually a wavelet transform is based on dilated-translated wavelets. We propose a symplectic-transformed-translated wavelet family psi(*)(r,s)(z-kappa) (r,s are the symplectic transform parameters, |s|(2)-|r|(2)=1, kappa is a translation parameter) generated from the mother wavelet psi and the corresponding wavelet transformation W(psi)f(r,s;kappa)=integral(infinity)(-infinity)(d(2)z/pi)f(z)psi(*)(r,s)(z-kappa). This new transform possesses well-behaved properties and is related to the optical Fresnel transform in quantum mechanical version. PMID:17099740

  19. Biolistic transformation of Scoparia dulcis L.

    PubMed

    Srinivas, Kota; Muralikrishna, Narra; Kumar, Kalva Bharath; Raghu, Ellendula; Mahender, Aileni; Kiranmayee, Kasula; Yashodahara, Velivela; Sadanandam, Abbagani

    2016-01-01

    Here, we report for the first time, the optimized conditions for microprojectile bombardment-mediated genetic transformation in Vassourinha (Scoparia dulcis L.), a Plantaginaceae medicinal plant species. Transformation was achieved by bombardment of axenic leaf segments with Binary vector pBI121 harbouring β-glucuronidase gene (GUS) as a reporter and neomycin phosphotransferase II gene (npt II) as a selectable marker. The influence of physical parameters viz., acceleration pressure, flight distance, gap width & macroprojectile travel distance of particle gun on frequency of transient GUS and stable (survival of putative transformants) expressions have been investigated. Biolistic delivery of the pBI121 yielded the best (80.0 %) transient expression of GUS gene bombarded at a flight distance of 6 cm and rupture disc pressure/acceleration pressure of 650 psi. Highest stable expression of 52.0 % was noticed in putative transformants on RMBI-K medium. Integration of GUS and npt II genes in the nuclear genome was confirmed through primer specific PCR. DNA blot analysis showed more than one transgene copy in the transformed plantlet genomes. The present study may be used for metabolic engineering and production of biopharmaceuticals by transplastomic technology in this valuable medicinal plant. PMID:27186019

  20. 8,9-DIHYDROXY-8,9-DIHYDRODIBENZO[A,L]PYRENE IS A POTENT MORPHOLOGICAL CELL-TRANSFORMING AGENT IN C3H10T1/2C18 MOUSE EMBRYO FIBROBLASTS IN THE ABSENCE OF DETECTABLE STABLE COVALENT DNA ADDUCTS

    EPA Science Inventory

    The comparative genotoxic effects of racemic trans-8,9dihydroxy-8,9-dihydrodibenzo[a,l]pyrene (trans- DB[a,l]P8,9-diol), the metabolic K-region dihydrodiol of dibenzo[a,l] pyrene (DB[a,l]P) (dibenzo[def,p]chrysene) and DB[a,l]P in transformable mouse embryo C3HIOT1/2C18 (C3HIOT1/...

  1. Multi-stable cylindrical lattices

    NASA Astrophysics Data System (ADS)

    Pirrera, Alberto; Lachenal, Xavier; Daynes, Stephen; Weaver, Paul M.; Chenchiah, Isaac V.

    2013-11-01

    We present a cylindrical lattice structure that mimics the behaviour of the virus bacteriophage T4 in having two (or more) stable states which differ in their radii and length. While the virus achieves bistability through molecular mechanisms we use composite materials to exploit the interplay between pre-stress, material properties and structural geometry. We demonstrate (computationally) that multi-stability is a robust phenomenon. We also show (analytically) that it is possible to choose the design variables so that the energy is independent of the radius, thus resulting in every state of the structure being stable.

  2. Trans-mediastinal gunshot wounds: are "stable" patients really stable?

    PubMed

    Nagy, Kimberly K; Roberts, Roxanne R; Smith, Robert F; Joseph, Kimberly T; An, Gary C; Bokhari, Faran; Barrett, John

    2002-10-01

    Gunshot wounds that traverse the mediastinum frequently cause serious injury to the cardiac, vascular, pulmonary, and digestive structures contained within. Most patients present with unstable vital signs signifying the need for emergency operation. An occasional patient will present with stable vital signs. Work-ups for such a patient may range from surgical exploration to radiographic and endoscopic testing to mere observation. We report our experience with diagnostic work-up of the stable patient with a transmediastinal gunshot wound. All stable patients who present to our urban level I trauma center following a transmediastinal gunshot wound undergo diagnostic work-up consisting of chest radiograph, cardiac ultrasound, angiography, esophagoscopy, barium swallow, and bronchoscopy. The work-up is dependent on the trajectory of the missile. Information on these patients is kept in a prospective database maintained by the trauma attending physicians. This database was analyzed and comparisons were made using Student's t-test and the Fisher exact c2 as appropriate. Over a 68-month period, 50 stable patients were admitted following a transmediastinal gunshot wound. All of these patients had a chest radiograph followed by one or more of the above tests. 8 patients (16%) were found to have a mediastinal injury (4 cardiac, 3 vascular, and 1 tracheo-esophageal) requiring urgent operation (group 1). The remaining 42 patients (84%) did not have a mediastinal injury (group 2). There was no difference between groups with respect to blood pressure, pulse, respiratory rate, pH, base deficit, or initial chest tube output. There was one death in each group, and three complications in group 2. Patients may appear stable following a transmediastinal gunshot wound, even when they have life-threatening injuries. There is no difference in vital signs, blood gas, or hemothorax to indicate which patients have serious injuries. We advocate continued aggressive work-up of these patients

  3. 28-Channel rotary transformer

    NASA Technical Reports Server (NTRS)

    Mclyman, W. T.

    1981-01-01

    Transformer transmits power and digital data across rotating interface. Array has many parallel data channels, each with potential l megabaud data rate. Ferrite-cored transformers are spaced along rotor; airgap between them reduces crosstalk.

  4. Equations For Rotary Transformers

    NASA Technical Reports Server (NTRS)

    Salomon, Phil M.; Wiktor, Peter J.; Marchetto, Carl A.

    1988-01-01

    Equations derived for input impedance, input power, and ratio of secondary current to primary current of rotary transformer. Used for quick analysis of transformer designs. Circuit model commonly used in textbooks on theory of ac circuits.

  5. Chemical Transformation Simulator

    EPA Science Inventory

    The Chemical Transformation Simulator (CTS) is a web-based, high-throughput screening tool that automates the calculation and collection of physicochemical properties for an organic chemical of interest and its predicted products resulting from transformations in environmental sy...

  6. Transformation of Pasteurella novicida

    PubMed Central

    Tyeryar, Franklin J.; Lawton, William D.

    1969-01-01

    Deoxyribonucleic acid from a streptomycin-resistant mutant of Pasteurella novicida transformed portions of P. novicida streptomycin-sensitive populations to streptomycin-resistant. Similarly, mutants auxotrophic for tryptophan or purine biosynthesis were also transformed to nutritional independence. PMID:5359612

  7. Shiftable multiscale transforms

    NASA Technical Reports Server (NTRS)

    Simoncelli, Eero P.; Freeman, William T.; Adelson, Edward H.; Heeger, David J.

    1992-01-01

    A type of translational invariance, referred to as shiftability, is defined for wavelet transforms. The property of shiftability is first discussed with respect to individual parameters: spatial position, orientation, and scale. The discussion then focuses on transformations that are simultaneously shiftable with respect to subsets of these parameters. It is shown that the critical sampling condition on the wavelet transforms must be relaxed to achieve shiftability. Two example transforms are implemented and applied to several signal and image processing problems.

  8. Synthesis of thermally stable polymers

    NASA Technical Reports Server (NTRS)

    Butler, G. B.

    1978-01-01

    The reaction of bis triazo linediones with divinyl esters and substituted styrenes was investigated. Twenty new polymers were derived via reaction of two previously synthesized bis triazol linediones and four new bis atriazol linediones with eight styrenes. The structure and polymer properties of these thermally stable polymers was examined. The reaction of triazo linediones with enol esters was also considered.

  9. Bayesian stable isotope mixing models

    EPA Science Inventory

    In this paper we review recent advances in Stable Isotope Mixing Models (SIMMs) and place them into an over-arching Bayesian statistical framework which allows for several useful extensions. SIMMs are used to quantify the proportional contributions of various sources to a mixtur...

  10. Stable interactions via proper deformations

    NASA Astrophysics Data System (ADS)

    Kaparulin, D. S.; Lyakhovich, S. L.; Sharapov, A. A.

    2016-04-01

    A new method is proposed for switching on interactions that are compatible with global symmetries and conservation laws of the original free theory. The method is applied to the control of stability in Lagrangian and non-Lagrangian theories with higher derivatives. By way of illustration, a wide class of stable interactions is constructed for the Pais-Uhlenbeck oscillator.

  11. Stable Black Families. Final Report.

    ERIC Educational Resources Information Center

    Gary, Lawrence E.; And Others

    This document is the final report of a study conducted to determine what factors contribute to strong Black family life and how these strong families solve problems, in order to add to the knowledge base on stable families so as to enhance practical intervention with families in need, and to identify models of self-help strategies used by stable…

  12. Achieving Perspective Transformation.

    ERIC Educational Resources Information Center

    Nowak, Jens

    Perspective transformation is a consciously achieved state in which the individual's perspective on life is transformed. The new perspective serves as a vantage point for life's actions and interactions, affecting the way life is lived. Three conditions are basic to achieving perspective transformation: (1) "feeling" experience, i.e., getting in…

  13. Note: Tesla transformer damping

    NASA Astrophysics Data System (ADS)

    Reed, J. L.

    2012-07-01

    Unexpected heavy damping in the two winding Tesla pulse transformer is shown to be due to small primary inductances. A small primary inductance is a necessary condition of operability, but is also a refractory inefficiency. A 30% performance loss is demonstrated using a typical "spiral strip" transformer. The loss is investigated by examining damping terms added to the transformer's governing equations. A significant alteration of the transformer's architecture is suggested to mitigate these losses. Experimental and simulated data comparing the 2 and 3 winding transformers are cited to support the suggestion.

  14. Stable continuous-time autoregressive process driven by stable subordinator

    NASA Astrophysics Data System (ADS)

    Wyłomańska, Agnieszka; Gajda, Janusz

    2016-02-01

    In this paper we examine the continuous-time autoregressive moving average process driven by α-stable Lévy motion delayed by inverse stable subordinator. This process can be applied to high-frequency data with visible jumps and so-called "trapping-events". Those properties are often visible in financial time series but also in amorphous semiconductors, technical data describing the rotational speed of a machine working under various load regimes or data related to indoor air quality. We concentrate on the main characteristics of the examined subordinated process expressed in the language of the measures of dependence which are main tools used in statistical investigation of real data. However, because the analyzed system is based on the α-stable distribution therefore we cannot consider here the correlation (or covariance) as a main measure which indicates at the dependence inside the process. In the paper we examine the codifference, the more general measure of dependence defined for wide class of processes. Moreover we present the simulation procedure of the considered system and indicate how to estimate its parameters. The theoretical results we illustrate by the simulated data analysis.

  15. Cassava (Manihot esculenta Crantz).

    PubMed

    Msikita, Weston; Ihemere, Uzoma; Siritunga, Dimuth; Sayre, Richard T

    2006-01-01

    During the last three years the generation of stably transformed cassava plants having value-added traits has become a reality. Currently, two Agrobacterium-mediated transformation systems are routinely used to engineer cassava. These systems use either somatic embryos or friable embryogenic calli. This paper presents detailed protocols for the transformation of cassava using primary somatic embryos. The effects of explant types, tissue culture conditions, and bacterial and plasmid related factors on transformation efficiency are discussed. PMID:17033047

  16. Phase stable RF transport system

    DOEpatents

    Curtin, Michael T.; Natter, Eckard F.; Denney, Peter M.

    1992-01-01

    An RF transport system delivers a phase-stable RF signal to a load, such as an RF cavity of a charged particle accelerator. A circuit generates a calibration signal at an odd multiple frequency of the RF signal where the calibration signal is superimposed with the RF signal on a common cable that connects the RF signal with the load. Signal isolating diplexers are located at both the RF signal source end and load end of the common cable to enable the calibration to be inserted and extracted from the cable signals without any affect on the RF signal. Any phase shift in the calibration signal during traverse of the common cable is then functionally related to the phase shift in the RF signal. The calibration phase shift is used to control a phase shifter for the RF signal to maintain a stable RF signal at the load.

  17. Stable maps and Quot schemes

    NASA Astrophysics Data System (ADS)

    Popa, Mihnea; Roth, Mike

    2003-06-01

    In this paper we study the relationship between two different compactifications of the space of vector bundle quotients of an arbitrary vector bundle on a curve. One is Grothendieck's Quot scheme, while the other is a moduli space of stable maps to the relative Grassmannian. We establish an essentially optimal upper bound on the dimension of the two compactifications. Based on that, we prove that for an arbitrary vector bundle, the Quot schemes of quotients of large degree are irreducible and generically smooth. We precisely describe all the vector bundles for which the same thing holds in the case of the moduli spaces of stable maps. We show that there are in general no natural morphisms between the two compactifications. Finally, as an application, we obtain new cases of a conjecture on effective base point freeness for pluritheta linear series on moduli spaces of vector bundles.

  18. Treatment of stable angina pectoris.

    PubMed

    Palaniswamy, Chandrasekar; Aronow, Wilbert S

    2011-09-01

    Management of stable angina pectoris includes antianginal medications, medications to prevent progression of atherosclerosis, and aggressive treatment of causative risk factors. Antianginal medications commonly used include nitrates, beta-blockers, calcium channel blockers, and ranolazine. Antiplatelet agents, statins, and angiotensin-converting enzyme inhibitors are used in patients with these problems to prevent progression of atherosclerosis and/or premature cardiovascular death. Aggressive risk factor control with diet; exercise; treatment of diabetes, hypertension, and dyslipidemia; and strategies to stop smoking and reduce weight should be a part of treatment strategy in all patients. Patients with stable angina who have symptoms refractory to medical treatment usually require coronary angiography, followed by either percutaneous or surgical revascularization. Recent mechanical techniques for the treatment of refractory angina include transmyocardial laser revascularization, enhanced external counterpulsation, and spinal cord stimulation. PMID:20861717

  19. Advanced Thermally Stable Jet Fuels

    SciTech Connect

    A. Boehman; C. Song; H. H. Schobert; M. M. Coleman; P. G. Hatcher; S. Eser

    1998-01-01

    The Penn State program in advanced thermally stable jet fuels has five components: 1) development of mechanisms of degradation and solids formation; 2) quantitative measurement of growth of sub-micrometer and micrometer-sized particles during thermal stressing; 3) characterization of carbonaceous deposits by various instrumental and microscopic methods; 4) elucidation of the role of additives in retarding the formation of carbonaceous solids; and 5) assessment of the potential of producing high yields of cycloalkanes and hydroaromatics from coal.

  20. LP based approach to optimal stable matchings

    SciTech Connect

    Teo, Chung-Piaw; Sethuraman, J.

    1997-06-01

    We study the classical stable marriage and stable roommates problems using a polyhedral approach. We propose a new LP formulation for the stable roommates problem. This formulation is non-empty if and only if the underlying roommates problem has a stable matching. Furthermore, for certain special weight functions on the edges, we construct a 2-approximation algorithm for the optimal stable roommates problem. Our technique uses a crucial geometry of the fractional solutions in this formulation. For the stable marriage problem, we show that a related geometry allows us to express any fractional solution in the stable marriage polytope as convex combination of stable marriage solutions. This leads to a genuinely simple proof of the integrality of the stable marriage polytope. Based on these ideas, we devise a heuristic to solve the optimal stable roommates problem. The heuristic combines the power of rounding and cutting-plane methods. We present some computational results based on preliminary implementations of this heuristic.

  1. Modeling stable isotope and organic carbon in hillslope stormflow

    NASA Astrophysics Data System (ADS)

    Dusek, Jaromir; Vogel, Tomas; Dohnal, Michal; Marx, Anne; Jankovec, Jakub; Sanda, Martin; Votrubova, Jana; Barth, Johannes A. C.; Cislerova, Milena

    2016-04-01

    Reliable prediction of water movement and fluxes of dissolved substances (such as stable isotopes and organic carbon) at both the hillslope and the catchment scales remains a challenge due to complex boundary conditions and soil spatial heterogeneity. In addition, microbially mediated transformations of dissolved organic carbon (DOC) are known to affect balance of DOC in soils, hence the transformations need to be included in a conceptual model of a DOC transport. So far, only few studies utilized stable isotope information in modeling and even fewer linked dissolved carbon fluxes to mixing and/or transport models. In this study, stormflow dynamics of oxygen-18 isotope and dissolved organic carbon was analyzed using a physically based modeling approach. One-dimensional dual-continuum vertical flow and transport model, based on Richards and advection-dispersion equations, was used to simulate the subsurface transport processes in a forest soil during several observed rainfall-runoff episodes. The transport of heat in the soil profile was described by conduction-advection equation. Water flow and transport of solutes and heat were assumed to take place in two mutually communicating porous domains, the soil matrix and the network of preferential pathways. The rate of microbial transformations of DOC was assumed to depend on soil water content and soil temperature. Oxygen-18 and dissolved organic carbon concentrations were observed in soil pore water, hillslope stormflow (collected in the experimental hillslope trench), and stream discharge (at the catchment outlet). The modeling was used to analyze the transformation of input solute signals into output hillslope signals observed in the trench stormflow. Signatures of oxygen-18 isotope in hillslope stormflow as well as isotope concentration in soil pore water were predicted reasonably well. Due to complex nature of microbial transformations, prediction of DOC rate and transport was associated with a high uncertainty.

  2. How Stable Is Stable? Function versus Community Composition

    PubMed Central

    Fernández, Ana; Huang, Suiying; Seston, Sherry; Xing, Jian; Hickey, Robert; Criddle, Craig; Tiedje, James

    1999-01-01

    The microbial community dynamics of a functionally stable, well-mixed, methanogenic reactor fed with glucose were analyzed over a 605-day period. The reactor maintained constant pH and chemical oxygen demand removal during this period. Thirty-six rrn clones from each of seven sampling events were analyzed by amplified ribosomal DNA restriction analysis (ARDRA) for the Bacteria and Archaea domains and by sequence analysis of dominant members of the community. Operational taxonomic units (OTUs), distinguished as unique ARDRA patterns, showed reproducible distribution for three sample replicates. The highest diversity was observed in the Bacteria domain. The 16S ribosomal DNA Bacteria clone library contained 75 OTUs, with the dominant OTU accounting for 13% of the total clones, but just 21 Archaea OTUs were found, and the most prominent OTU represented 50% of the clones from the respective library. Succession in methanogenic populations was observed, and two periods were distinguished: in the first, Methanobacterium formicicum was dominant, and in the second, Methanosarcina mazei and a Methanobacterium bryantii-related organism were dominant. Higher variability in Bacteria populations was detected, and the temporal OTU distribution suggested a chaotic pattern. Although dominant OTUs were constantly replaced from one sampling point to the next, phylogenetic analysis indicated that inferred physiologic changes in the community were not as dramatic as were genetic changes. Seven of eight dominant OTUs during the first period clustered with the spirochete group, although a cyclic pattern of substitution occurred among members within this order. A more flexible community structure characterized the second period, since a sequential replacement of a Eubacterium-related organism by an unrelated deep-branched organism and finally by a Propionibacterium-like species was observed. Metabolic differences among the dominant fermenters detected suggest that changes in carbon and

  3. Foraging and farming as niche construction: stable and unstable adaptations

    PubMed Central

    Rowley-Conwy, Peter; Layton, Robert

    2011-01-01

    All forager (or hunter–gatherer) societies construct niches, many of them actively by the concentration of wild plants into useful stands, small-scale cultivation, burning of natural vegetation to encourage useful species, and various forms of hunting, collectively termed ‘low-level food production’. Many such niches are stable and can continue indefinitely, because forager populations are usually stable. Some are unstable, but these usually transform into other foraging niches, not geographically expansive farming niches. The Epipalaeolithic (final hunter–gatherer) niche in the Near East was complex but stable, with a relatively high population density, until destabilized by an abrupt climatic change. The niche was unintentionally transformed into an agricultural one, due to chance genetic and behavioural attributes of some wild plant and animal species. The agricultural niche could be exported with modifications over much of the Old World. This was driven by massive population increase and had huge impacts on local people, animals and plants wherever the farming niche was carried. Farming niches in some areas may temporarily come close to stability, but the history of the last 11 000 years does not suggest that agriculture is an effective strategy for achieving demographic and political stability in the world's farming populations. PMID:21320899

  4. Construction of energy-stable Galerkin reduced order models.

    SciTech Connect

    Kalashnikova, Irina; Barone, Matthew Franklin; Arunajatesan, Srinivasan; van Bloemen Waanders, Bart Gustaaf

    2013-05-01

    This report aims to unify several approaches for building stable projection-based reduced order models (ROMs). Attention is focused on linear time-invariant (LTI) systems. The model reduction procedure consists of two steps: the computation of a reduced basis, and the projection of the governing partial differential equations (PDEs) onto this reduced basis. Two kinds of reduced bases are considered: the proper orthogonal decomposition (POD) basis and the balanced truncation basis. The projection step of the model reduction can be done in two ways: via continuous projection or via discrete projection. First, an approach for building energy-stable Galerkin ROMs for linear hyperbolic or incompletely parabolic systems of PDEs using continuous projection is proposed. The idea is to apply to the set of PDEs a transformation induced by the Lyapunov function for the system, and to build the ROM in the transformed variables. The resulting ROM will be energy-stable for any choice of reduced basis. It is shown that, for many PDE systems, the desired transformation is induced by a special weighted L2 inner product, termed the %E2%80%9Csymmetry inner product%E2%80%9D. Attention is then turned to building energy-stable ROMs via discrete projection. A discrete counterpart of the continuous symmetry inner product, a weighted L2 inner product termed the %E2%80%9CLyapunov inner product%E2%80%9D, is derived. The weighting matrix that defines the Lyapunov inner product can be computed in a black-box fashion for a stable LTI system arising from the discretization of a system of PDEs in space. It is shown that a ROM constructed via discrete projection using the Lyapunov inner product will be energy-stable for any choice of reduced basis. Connections between the Lyapunov inner product and the inner product induced by the balanced truncation algorithm are made. Comparisons are also made between the symmetry inner product and the Lyapunov inner product. The performance of ROMs constructed

  5. Will it form a stable glass? How the stability of vapor deposited glasses depends on molecular structure

    NASA Astrophysics Data System (ADS)

    Tylinski, Michael; Beasley, Madeleine; Chua, Yeong Zen; Schick, Christoph; Ediger, Mark

    Over the past nine years physical vapor deposition has been used to prepare molecular glasses with exceptional properties. When heated, transformation of these highly stable glasses takes orders of magnitude longer than the transformation of liquid-cooled glasses. Until recently, it appeared that most organic molecules could form stable glasses when vapor deposited. We test the generality of stable glass formation by vapor-depositing a wide range of small organic molecules, including hydroxyl, carbonyl, phosphate, aromatic, and aliphatic functional groups. When prepared under conditions expected to yield highly stable glasses, we observe glasses with a wide range of kinetic stabilities, depending on the functional groups in the molecule. In general, alcohols and molecules with long aliphatic chains do not form highly stable glasses while aromatic molecules do. We also test the hypothesis that the surface mobility during deposition determines if a molecule is able to create highly stable glasses.

  6. Perception of shape and space across rigid transformations.

    PubMed

    Schmidt, Filipp; Spröte, Patrick; Fleming, Roland W

    2016-09-01

    Objects in our environment are subject to manifold transformations, either of the physical objects themselves or of the object images on the retina. Despite drastic effects on the objects' physical appearances, we are often able to identify stable objects across transformations and have strong subjective impressions of the transformations themselves. This suggests the brain is equipped with sophisticated mechanisms for inferring both object constancy, and objects' causal history. We employed a dot-matching task to study in geometrical detail the effects of rigid transformations on representations of shape and space. We presented an untransformed 'base shape' on the left side of the screen and its transformed counterpart on the right (rotated, scaled, or both). On each trial, a dot was superimposed at a given location on the contour (Experiment 1) or within and around the shape (Experiment 2). The participant's task was to place a dot at the corresponding location on the right side of the screen. By analyzing correspondence between responses and physical transformations, we tested for object constancy, causal history, and transformation of space. We find that shape representations are remarkably robust against rotation and scaling. Performance is modulated by the type and amount of transformation, as well as by contour saliency. We also find that the representation of space within and around a shape is transformed in line with the shape transformation, as if shape features establish an object-centered reference frame. These findings suggest robust mechanisms for the inference of shape, space and correspondence across transformations. PMID:25937375

  7. Catalytic coherence transformations

    NASA Astrophysics Data System (ADS)

    Bu, Kaifeng; Singh, Uttam; Wu, Junde

    2016-04-01

    Catalytic coherence transformations allow the otherwise impossible state transformations using only incoherent operations with the aid of an auxiliary system with finite coherence that is not being consumed in any way. Here we find the necessary and sufficient conditions for the deterministic and stochastic catalytic coherence transformations between a pair of pure quantum states. In particular, we show that the simultaneous decrease of a family of Rényi entropies of the diagonal parts of the states under consideration is a necessary and sufficient condition for the deterministic catalytic coherence transformations. Similarly, for stochastic catalytic coherence transformations we find the necessary and sufficient conditions for achieving a higher optimal probability of conversion. We thus completely characterize the coherence transformations among pure quantum states under incoherent operations. We give numerous examples to elaborate our results. We also explore the possibility of the same system acting as a catalyst for itself and find that indeed self-catalysis is possible. Further, for the cases where no catalytic coherence transformation is possible we provide entanglement-assisted coherence transformations and find the necessary and sufficient conditions for such transformations.

  8. Magnetically Controlled Variable Transformer

    NASA Technical Reports Server (NTRS)

    Kleiner, Charles T.

    1994-01-01

    Improved variable-transformer circuit, output voltage and current of which controlled by use of relatively small current supplied at relatively low power to control windings on its magnetic cores. Transformer circuits of this type called "magnetic amplifiers" because ratio between controlled output power and power driving control current of such circuit large. This ratio - power gain - can be as large as 100 in present circuit. Variable-transformer circuit offers advantages of efficiency, safety, and controllability over some prior variable-transformer circuits.

  9. Biomolecular surface construction by PDE transform.

    PubMed

    Zheng, Qiong; Yang, Siyang; Wei, Guo-Wei

    2012-03-01

    virus surface capsid. Virus surface morphologies of different resolutions are attained by adjusting the propagation time. Therefore, the present PDE transform provides a multiresolution analysis in the surface visualization. Extensive numerical experiment and comparison with an established surface model indicate that the present PDE transform is a robust, stable, and efficient approach for biomolecular surface generation in Cartesian meshes. PMID:22582140

  10. Stable Hemiaminals: 2-Aminopyrimidine Derivatives.

    PubMed

    Kwiecień, Anna; Ciunik, Zbigniew

    2015-01-01

    Stable hemiaminals can be obtained in the one-pot reaction between 2-aminopyrimidine and nitrobenzaldehyde derivatives. Ten new hemiaminals have been obtained, six of them in crystal state. The molecular stability of these intermediates results from the presence of both electron-withdrawing nitro groups as substituents on the phenyl ring and pyrimidine ring, so no further stabilisation by intramolecular interaction is required. Hemiaminal molecules possess a tetrahedral carbon atom constituting a stereogenic centre. As the result of crystallisation in centrosymmetric space groups both enantiomers are present in the crystal structure. PMID:26258772

  11. Field performance of transgenic sugarcane produced using Agrobacterium and biolistics methods.

    PubMed

    Joyce, Priya; Hermann, Scott; O'Connell, Anthony; Dinh, Quang; Shumbe, Leonard; Lakshmanan, Prakash

    2014-05-01

    Future genetic improvement of sugarcane depends, in part, on the ability to produce high-yielding transgenic cultivars with improved traits such as herbicide and insect resistance. Here, transgenic sugarcane plants generated by different transformation methods were assessed for field performance over 3 years. Agrobacterium-mediated (Agro) transgenic events (35) were produced using four different Agrobacterium tumefaciens strains, while biolistic (Biol) transgenic events (48) were produced using either minimal linearized DNA (LDNA) transgene cassettes with 5', 3' or blunt ends or whole circular plasmid (PDNA) vectors containing the same transgenes. A combined analysis showed a reduction in growth and cane yield in Biol, Agro as well as untransformed tissue culture (TC) events, compared with the parent clone (PC) Q117 (no transformation or tissue culture) in the plant, first ratoon and second ratoon crops. However, when individual events were analysed separately, yields of some transgenic events from both Agro and Biol were comparable to PC, suggesting that either transformation method can produce commercially suitable clones. Interestingly, a greater percentage of Biol transformants were similar to PC for growth and yield than Agro clones. Crop ratoonability and sugar yield components (Brix%, Pol%, and commercial cane sugar (CCS)) were unaffected by transformation or tissue culture. Transgene expression remained stable over different crop cycles and increased with plant maturity. Transgene copy number did not influence transgene expression, and both transformation methods produced low transgene copy number events. No consistent pattern of genetic changes was detected in the test population using three DNA fingerprinting techniques. PMID:24330327

  12. How Physicists Made Stable Lévy Processes Physically Plausible

    NASA Astrophysics Data System (ADS)

    Schinckus, Christophe

    2013-08-01

    Stable Lévy processes have very interesting properties for describing the complex behaviour of non-equilibrium dissipative systems such as turbulence, anomalous diffusion or financial markets. However, although these processes better fit the empirical data, some of their statistical properties can raise several theoretical problems in empirical applications because they generate infinite variables. Econophysicists have developed statistical solutions to make these processes physically plausible. This paper presents a review of these analytical solutions (truncations) for stable Lévy processes and how econophysicists transformed them into data-driven processes. The evolution of these analytical solutions is presented as a progressive research programme provided by (econo)physicists for theoretical problems encountered in financial economics in the 1960s and the 1970s.

  13. Dielectric Relaxation of Materials that Form Ultra-Stable Glasses

    NASA Astrophysics Data System (ADS)

    Richert, Ranko

    2015-03-01

    Physical vapor deposition of glass forming materials onto substrates at temperatures around 0.8 Tg produces glasses of high density and low enthalpy. Using interdigitated electrode cells as substrates, such stable glasses can be studied by dielectric spectroscopy in situ. This technique is applied to monitor the dynamics of stable films upon their conversion to the ordinary supercooled liquid state. The dielectric loss during transformation indicates that the softening proceeds by a growth front mechanism and generates the ordinary liquid state without forming intermediates. The same technique is also used to assess the residual dynamics of the stable glassy state. We observe that processes such as the Johari-Goldstein beta relaxation are strongly suppressed in this stable state, consistent with the relatively low fictive temperature of these glassy states. coauthors: Hai-Bin Yu, Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85278; Michael Tylinski, and Mark D. Ediger, Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706.

  14. Persistence Length of Stable Microtubules

    NASA Astrophysics Data System (ADS)

    Hawkins, Taviare; Mirigian, Matthew; Yasar, M. Selcuk; Ross, Jennifer

    2011-03-01

    Microtubules are a vital component of the cytoskeleton. As the most rigid of the cytoskeleton filaments, they give shape and support to the cell. They are also essential for intracellular traffic by providing the roadways onto which organelles are transported, and they are required to reorganize during cellular division. To perform its function in the cell, the microtubule must be rigid yet dynamic. We are interested in how the mechanical properties of stable microtubules change over time. Some ``stable'' microtubules of the cell are recycled after days, such as in the axons of neurons or the cilia and flagella. We measured the persistence length of freely fluctuating taxol-stabilized microtubules over the span of a week and analyzed them via Fourier decomposition. As measured on a daily basis, the persistence length is independent of the contour length. Although measured over the span of the week, the accuracy of the measurement and the persistence length varies. We also studied how fluorescently-labeling the microtubule affects the persistence length and observed that a higher labeling ratio corresponded to greater flexibility. National Science Foundation Grant No: 0928540 to JLR.

  15. Stable Imaging for Astronomy (SIA)

    NASA Astrophysics Data System (ADS)

    Beaulieu, Mathilde; Ottogalli, Sebastien; Preis, Olivier; Bresson, Yves; Rivet, Jean-Pierre; Abe, Lyu; Vakili, Farrokh

    2014-07-01

    One of the most challenging fields of astronomical instrumentation is probably high-contrast imaging since it ultimately combines ultra-high sensitivity at low flux and the ability to cope with photon flux contrasts of several hundreds of millions or even more. These two aspects implicitly require that high-contrast instruments should be highly stable in the sense of the reproducibility of their measurements at different times, but also, continuously stable over time. In most high contrast instruments or experiments, their sensitivity is broken after at most tens of minutes of operation due to uncontrolled and unknown behaviour of the whole experiment regarding the environmental conditions. In this paper, we introduce a general approach of an exhaustive stability study for high-contrast imaging that has been initiated at Lagrange Laboratory, Observatoire de la Côte d'Azur (OCA). On a practical ground, one of the fundamental issues of this study is the metrology, which is the basis of all reproducible measurements. We describe a small experiment designed to understand the behaviour of one of our ultra-precise metrology tools (a commercial sub-nanometric 3-way interferometer) and derive the conditions under which its operation delivers reliable results. The approach will apply to the high-contrast imaging test-bench SPEED, under development at OCA.

  16. Transformer design tradeoffs

    NASA Technical Reports Server (NTRS)

    Mclyman, W. T.

    1977-01-01

    Technical memorandum includes transformer area product numbers, which are used to summarize dimensional and electrical properties of C-cores, pot cores, lamination, powder cores, and tape-wound cores. To aid in core selection, comparison of five common core materials is presented to indicate their influence on overall transformer efficiency and weight.

  17. Biochemical transformation of coals

    DOEpatents

    Lin, M.S.; Premuzic, E.T.

    1999-03-23

    A method of biochemically transforming macromolecular compounds found in solid carbonaceous materials, such as coal is provided. The preparation of new microorganisms, metabolically weaned through challenge growth processes to biochemically transform solid carbonaceous materials at extreme temperatures, pressures, pH, salt and toxic metal concentrations is also disclosed. 7 figs.

  18. Fourier Transform Spectrometer

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Understanding the global atmospheric changes is difficult with today's current technology. However, with high resolution and nearly continuous observations from a satellite, it's possible to transform our understanding of the atmosphere. To enable the next generation of atmospheric science, a new class of orbiting atmospheric sensors is being developed. The foundation of this advanced concept is the Fourier Transform Spectrometer, or FTS.

  19. Deployment & Market Transformation (Brochure)

    SciTech Connect

    Not Available

    2012-04-01

    NREL's deployment and market transformation (D and MT) activities encompass the laboratory's full range of technologies, which span the energy efficiency and renewable energy spectrum. NREL staff educates partners on how they can advance sustainable energy applications and also provides clients with best practices for reducing barriers to innovation and market transformation.

  20. Genetic Transformation of Bacteria.

    ERIC Educational Resources Information Center

    Moss, Robert.

    1991-01-01

    An activity in which students transform an ampicillin-sensitive strain of E. coli with a plasmid containing a gene for ampicillin resistance is described. The procedure for the preparation of competent cells and the transformation of competent E. coli is provided. (KR)

  1. Support Principals, Transform Schools

    ERIC Educational Resources Information Center

    Aguilar, Elena; Goldwasser, Davina; Tank-Crestetto, Kristina

    2011-01-01

    The Transformational Coaching Team in Oakland Unified School District provides differentiated, sustained, job-embedded support to the district's school leaders. In this article, members of the team describe how they work with principals to transform the culture of schools. Student achievement data show above-average improvement in schools in which…

  2. Two Different Squeeze Transformations

    NASA Technical Reports Server (NTRS)

    Han, D. (Editor); Kim, Y. S.

    1996-01-01

    Lorentz boosts are squeeze transformations. While these transformations are similar to those in squeezed states of light, they are fundamentally different from both physical and mathematical points of view. The difference is illustrated in terms of two coupled harmonic oscillators, and in terms of the covariant harmonic oscillator formalism.

  3. A Transformation Called "Twist"

    ERIC Educational Resources Information Center

    Hwang, Daniel

    2010-01-01

    The transformations found in secondary mathematics curriculum are typically limited to stretches and translations (e.g., ACARA, 2010). Advanced students may find the transformation, twist, to be of further interest. As most available resources are written for professional-level readers, this article is intended to be an introduction accessible to…

  4. Disc piezoelectric ceramic transformers.

    PubMed

    Erhart, Jirií; Půlpán, Petr; Doleček, Roman; Psota, Pavel; Lédl, Vít

    2013-08-01

    In this contribution, we present our study on disc-shaped and homogeneously poled piezoelectric ceramic transformers working in planar-extensional vibration modes. Transformers are designed with electrodes divided into wedge, axisymmetrical ring-dot, moonie, smile, or yin-yang segments. Transformation ratio, efficiency, and input and output impedances were measured for low-power signals. Transformer efficiency and transformation ratio were measured as a function of frequency and impedance load in the secondary circuit. Optimum impedance for the maximum efficiency has been found. Maximum efficiency and no-load transformation ratio can reach almost 100% and 52 for the fundamental resonance of ring-dot transformers and 98% and 67 for the second resonance of 2-segment wedge transformers. Maximum efficiency was reached at optimum impedance, which is in the range from 500 Ω to 10 kΩ, depending on the electrode pattern and size. Fundamental vibration mode and its overtones were further studied using frequency-modulated digital holographic interferometry and by the finite element method. Complementary information has been obtained by the infrared camera visualization of surface temperature profiles at higher driving power. PMID:25004532

  5. Transformative Learning and Identity

    ERIC Educational Resources Information Center

    Illeris, Knud

    2014-01-01

    Transformative learning has usually been defined as transformations of meaning perspectives, frames of reference, and habits of mind--as proposed initially by Jack Mezirow. However, several authors have found this definition too narrow and too cognitively oriented, and Mezirow has later emphasized that emotional and social conditions are also…

  6. Adaptive Wavelet Transforms

    SciTech Connect

    Szu, H.; Hsu, C.

    1996-12-31

    Human sensors systems (HSS) may be approximately described as an adaptive or self-learning version of the Wavelet Transforms (WT) that are capable to learn from several input-output associative pairs of suitable transform mother wavelets. Such an Adaptive WT (AWT) is a redundant combination of mother wavelets to either represent or classify inputs.

  7. Direct current transformer

    NASA Technical Reports Server (NTRS)

    Khanna, S. M.; Urban, E. W. (Inventor)

    1979-01-01

    A direct current transformer in which the primary consists of an elongated strip of superconductive material, across the ends of which is direct current potential is described. Parallel and closely spaced to the primary is positioned a transformer secondary consisting of a thin strip of magnetoresistive material.

  8. Stable microbial community composition on the Greenland Ice Sheet.

    PubMed

    Musilova, Michaela; Tranter, Martyn; Bennett, Sarah A; Wadham, Jemma; Anesio, Alexandre M

    2015-01-01

    The first molecular-based studies of microbes in snow and on glaciers have only recently been performed on the vast Greenland Ice Sheet (GrIS). Aeolian microbial seeding is hypothesized to impact on glacier surface community compositions. Localized melting of glacier debris (cryoconite) into the surface ice forms cryoconite holes, which are considered 'hot spots' for microbial activity on glaciers. To date, few studies have attempted to assess the origin and evolution of cryoconite and cryoconite hole communities throughout a melt season. In this study, a range of experimental approaches was used for the first time to study the inputs, temporal and structural transformations of GrIS microbial communities over the course of a whole ablation season. Small amounts of aeolian (wind and snow) microbes were potentially seeding the stable communities that were already present on the glacier (composed mainly of Proteobacteria, Cyanobacteria, and Actinobacteria). However, the dominant bacterial taxa in the aeolian samples (Firmicutes) did not establish themselves in local glacier surface communities. Cryoconite and cryoconite hole community composition remained stable throughout the ablation season following the fast community turnover, which accompanied the initial snow melt. The presence of stable communities in cryoconite and cryoconite holes on the GrIS will allow future studies to assess glacier surface microbial diversity at individual study sites from sampling intervals of short duration only. Aeolian inputs also had significantly different organic δ(13)C values (-28.0 to -27.0‰) from the glacier surface values (-25.7 to -23.6‰), indicating that in situ microbial processes are important in fixing new organic matter and transforming aeolian organic carbon. The continuous productivity of stable communities over one melt season makes them important contributors to biogeochemical nutrient cycling on glaciers. PMID:25852658

  9. The stable and metastable Ti-Nb phase diagrams

    NASA Astrophysics Data System (ADS)

    Moffat, D. L.; Kattner, U. R.

    1988-10-01

    The phase transformations which occur in the Ti-Nb binary alloy system have been discussed in two recent papers. The phase relationships were investigated by varying alloy composition and thermal history. In this paper, these results are summarized in complete and thermodynamically consistent calculations of the stable and metastable phase diagrams. The calculations of the metastable equilibria are relevant to the Ti-V and Ti-Mo systems, as well as to several other titanium and zirconium-based transition metal alloy systems.

  10. Formation of stable chlorinated hydrocarbons in weathering plant material

    SciTech Connect

    Myneni, Satish C. B.

    2001-10-01

    Though several chlorinated organic compounds produced by humans are carcinogenic and toxic, some are also produced by the biotic and abiotic processes in the environment. In situ x-ray spectroscopy data indicate that natural organic matter in soils, sediments, and natural waters contain stable, less volatile organic compounds with chlorinated phenolic and aliphatic groups as the principal Cl forms. These compounds are formed at rapid rates from the transformation of inorganic Cl during humification of plant material and, thus, play a critical role in the cycling of Cl and of several major and trace elements in the environment and may influence human health. [References: 19

  11. Stable Algorithm For Estimating Airdata From Flush Surface Pressure Measurements

    NASA Technical Reports Server (NTRS)

    Whitmore, Stephen, A. (Inventor); Cobleigh, Brent R. (Inventor); Haering, Edward A., Jr. (Inventor)

    2001-01-01

    An airdata estimation and evaluation system and method, including a stable algorithm for estimating airdata from nonintrusive surface pressure measurements. The airdata estimation and evaluation system is preferably implemented in a flush airdata sensing (FADS) system. The system and method of the present invention take a flow model equation and transform it into a triples formulation equation. The triples formulation equation eliminates the pressure related states from the flow model equation by strategically taking the differences of three surface pressures, known as triples. This triples formulation equation is then used to accurately estimate and compute vital airdata from nonintrusive surface pressure measurements.

  12. Mixture of Skewed α-Stable Distributions

    NASA Astrophysics Data System (ADS)

    Shojaei, S. R. Hosseini; Nassiri, V.; Mohammadian, Gh. R.; Mohammadpour, A.

    2011-03-01

    Expectation maximization (EM) algorithm and the Bayesian techniques are two approaches for statistical inference of mixture models [3, 4]. By noting the advantages of the Bayesian methods, practitioners prefer them. However, implementing Markov chain Monte Carlo algorithms can be very complicated for stable distributions, due to the non-analytic density or distribution function formulas. In this paper, we introduce a new class of mixture of heavy-tailed distributions, called mixture of skewed stable distributions. Skewed stable distributions belongs to the exponential family and they have analytic density representation. It is shown that skewed stable distributions dominate skew stable distribution functions and they can be used to model heavy-tailed data. The class of skewed stable distributions has an analytic representation for its density function and the Bayesian inference can be done similar to the exponential family of distributions. Finally, mixture of skewed stable distributions are compared to the mixture of stable distributions through a simulations study.

  13. Stable density stratification solar pond

    NASA Technical Reports Server (NTRS)

    Lansing, F. L. (Inventor)

    1985-01-01

    A stable density-stratification solar pond for use in the collection and storage of solar thermal energy including a container having a first section characterized by an internal wall of a substantially cylindrical configuration and a second section having an internal wall of a substantially truncated conical configuration surmounting the first section in coaxial alignment therewith, the second section of said container being characterized by a base of a diameter substantially equal to the diameter of the first section and a truncated apex defining a solar energy acceptance opening is discussed. A body of immiscible liquids is disposed within the container and comprises a lower portion substantially filling the first section of the container and an upper portion substantially filling the second section of the container, said lower portion being an aqueous based liquid of a darker color than the upper portion and of a greater density. A protective cover plate is removably provided for covering the acceptance opening.

  14. Stable massive particles at colliders

    SciTech Connect

    Fairbairn, M.; Kraan, A.C.; Milstead, D.A.; Sjostrand, T.; Skands, P.; Sloan, T.; /Lancaster U.

    2006-11-01

    We review the theoretical motivations and experimental status of searches for stable massive particles (SMPs) which could be sufficiently long-lived as to be directly detected at collider experiments. The discovery of such particles would address a number of important questions in modern physics including the origin and composition of dark matter in the universe and the unification of the fundamental forces. This review describes the techniques used in SMP-searches at collider experiments and the limits so far obtained on the production of SMPs which possess various colour, electric and magnetic charge quantum numbers. We also describe theoretical scenarios which predict SMPs, the phenomenology needed to model their production at colliders and interactions with matter. In addition, the interplay between collider searches and open questions in cosmology such as dark matter composition are addressed.

  15. Dimensionally stable metallic hydride composition

    DOEpatents

    Heung, Leung K.

    1994-01-01

    A stable, metallic hydride composition and a process for making such a composition. The composition comprises a uniformly blended mixture of a metal hydride, kieselguhr, and a ballast metal, all in the form of particles. The composition is made by subjecting a metal hydride to one or more hydrogen absorption/desorption cycles to disintegrate the hydride particles to less than approximately 100 microns in size. The particles are partly oxidized, then blended with the ballast metal and the kieselguhr to form a uniform mixture. The mixture is compressed into pellets and calcined. Preferably, the mixture includes approximately 10 vol. % or more kieselguhr and approximately 50 vol. % or more ballast. Metal hydrides that can be used in the composition include Zr, Ti, V, Nb, Pd, as well as binary, tertiary, and more complex alloys of La, Al, Cu, Ti, Co, Ni, Fe, Zr, Mg, Ca, Mn, and mixtures and other combinations thereof. Ballast metals include Al, Cu and Ni.

  16. Novel vortex-transform for high frequency modulated patterns.

    PubMed

    Sierra-Sosa, Daniel; Angel-Toro, Luciano; Bolognini, Nestor; Tebaldi, Myrian

    2013-10-01

    A novel vortex-transform is proposed. This transform allows for generating complex-valued functions from modulated intensity patterns, including high frequency components from modulation, without the generation of unstable phase singularities. From these complex-valued functions it is possible to obtain intensity and pseudo-phase maps to analyze the intensity recordings without the necessity of phase retrieval techniques. The intensity and pseudo-phase maps obtained by using this transform preserve the modulation structure onto the intensity and phase modulo 2π maps, including stable phase singularities. PMID:24104283

  17. Sugar feeding in adult stable flies

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Adult stable flies, (Stomoxys calcitrans (L.)), are known to feed readily on sugars in the laboratory. However, little is known concerning the extent of stable fly sugar feeding in wild populations. We examined the frequency of sugar feeding in stable flies in rural and urban environments. In additi...

  18. Uses of stable isotopes in fish ecology

    EPA Science Inventory

    Analyses of fish tissues (other than otoliths) for stable isotope ratios can provide substantial information on fish ecology, including physiological ecology. Stable isotopes of nitrogen and carbon frequently are used to determine the mix of diet sources for consumers. Stable i...

  19. Stable line defects in silicene

    NASA Astrophysics Data System (ADS)

    Ghosh, Dibyajyoti; Parida, Prakash; Pati, Swapan K.

    2015-11-01

    Line defects in two-dimensional (2D) materials greatly modulate various properties of their pristine form. Using ab initio molecular dynamics (AIMD) simulations, we investigate the structural reconstructions of different kinds of grain boundaries in the silicene sheets. It is evident that depending upon the presence of silicon adatoms and edge shape of grain boundaries (i.e., armchair or zigzag), stable extended line defects (ELDs) can be introduced in a controlled way. Further studies show the stability of these line-defects in silicene, grown on Ag(111) surface at room-temperature. Importantly, unlike most of the 2D sheet materials such as graphene and hexagonal boron nitride, 5-5-8 line defects modify the nonmagnetic semimetallic pristine silicene sheet to spin-polarized metal. As ferromagnetically ordered magnetic moments remain strongly localized at the line defect, a one-dimensional spin channel gets created in silicene. Interestingly, these spin channels are quite stable because, unlike the edge of nanoribbons, structural reconstruction or contamination cannot destroy the ordering of magnetic moments here. Zigzag silicene nanoribbons with a 5-5-8 line defect also exhibit various interesting electronic and magnetic properties depending upon their width as well as the nature of the magnetic coupling between edge and defect spin states. Upon incorporation of other ELDs, such as 4-4-4 and 4-8 defects, 2D sheets and nanoribbons of silicene show a nonmagnetic metallic or semiconducting ground state. Highlighting the controlled formation of ELDs and consequent emergence of technologically important properties in silicene, we propose new routes to realize silicene-based nanoelectronic and spintronic devices.

  20. Transformer design tradeoffs

    NASA Technical Reports Server (NTRS)

    Mclyman, W. T.

    1976-01-01

    Material was presented to assist transformer designers in the transition from long-used English units to the less familiar metric equivalents. A coordination between the area product numbers ap (product of window and core cross-section areas) and current density J was developed for a given regulation and temperature rise. Straight-line relationships for Ap and Volume, Ap and surface area At and, Ap and weight were developed. These relationships can now be used as new tools to simplify and standardize the process of transformer design. They also made it possible to design transformers of small bulk and volume or to optimize efficiency.

  1. Optical sine transformation.

    PubMed

    Yang, G; Zhang, J; Gong, J; Chen, J; Ho, Y

    1987-10-15

    The phase mask distribution of optical sine transformation (OST) has been calculated according to the optical general transformation theory. To avoid the diffraction loss of the phase mask, the optical waveguide method is used. Computation shows that the optical sine transformation is possible with only one phase mask, i.e., one-half of a cylindrical lens in the 1-D case and one-quarter of a spherical lens in the 2-D case. Experimental results agree with the theoretical prediction. Image compression by OST is also given. PMID:20523380

  2. Advances in the Natural transform

    NASA Astrophysics Data System (ADS)

    Belgacem, F. B. M.; Silambarasan, R.

    2012-11-01

    The literature review of the Natural transform and the existing definitions and connections to the Laplace and Sumudu transforms are discussed in this communication. Along with the complex inverse Natural transform and Heaviside's expansion formula, the relation of Bessel's function to Natural transform (and hence Laplace and Sumudu transforms) are defined.

  3. A discrete fractional random transform

    NASA Astrophysics Data System (ADS)

    Liu, Zhengjun; Zhao, Haifa; Liu, Shutian

    2005-11-01

    We propose a discrete fractional random transform based on a generalization of the discrete fractional Fourier transform with an intrinsic randomness. Such discrete fractional random transform inheres excellent mathematical properties of the fractional Fourier transform along with some fantastic features of its own. As a primary application, the discrete fractional random transform has been used for image encryption and decryption.

  4. A Classical Science Transformed.

    ERIC Educational Resources Information Center

    Kovalevsky, Jean

    1979-01-01

    Describes how satellites and other tools of space technology have transformed classical geodesy into the science of space geodynamics. The establishment and the activities of the French Center for Geodynamic and Astronomical Research Studies (CERGA) are also included. (HM)

  5. Transformer design tradeoffs

    NASA Technical Reports Server (NTRS)

    Mclyman, W. T.

    1977-01-01

    In space, power system transformer components are frequently the heaviest and bulkiest items in the power conversion circuit. They also have a significant effect upon the overall performance and efficiency of the system. Accordingly, the design of such transformers has an important effect on overall system weight, power-inversion efficiency, and cost. Relationships were between the parameters used by transformer designers that can be used as new tools to standardize and simplify transformer design. They can be used to optimize the design either for small size and weight or efficiency. The metric system of units, rather than the familiar English units, is used; however, material is presented to assist the reader in the transition from one system to the other.

  6. Imaging Fourier transform spectrometer

    SciTech Connect

    Bennett, C.L.

    1993-09-13

    This invention is comprised of an imaging Fourier transform spectrometer having a Fourier transform infrared spectrometer providing a series of images to a focal plane array camera. The focal plane array camera is clocked to a multiple of zero crossing occurrences as caused by a moving mirror of the Fourier transform infrared spectrometer and as detected by a laser detector such that the frame capture rate of the focal plane array camera corresponds to a multiple of the zero crossing rate of the Fourier transform infrared spectrometer. The images are transmitted to a computer for processing such that representations of the images as viewed in the light of an arbitrary spectral ``fingerprint`` pattern can be displayed on a monitor or otherwise stored and manipulated by the computer.

  7. Fourier Transform Mass Spectrometry

    PubMed Central

    Scigelova, Michaela; Hornshaw, Martin; Giannakopulos, Anastassios; Makarov, Alexander

    2011-01-01

    This article provides an introduction to Fourier transform-based mass spectrometry. The key performance characteristics of Fourier transform-based mass spectrometry, mass accuracy and resolution, are presented in the view of how they impact the interpretation of measurements in proteomic applications. The theory and principles of operation of two types of mass analyzer, Fourier transform ion cyclotron resonance and Orbitrap, are described. Major benefits as well as limitations of Fourier transform-based mass spectrometry technology are discussed in the context of practical sample analysis, and illustrated with examples included as figures in this text and in the accompanying slide set. Comparisons highlighting the performance differences between the two mass analyzers are made where deemed useful in assisting the user with choosing the most appropriate technology for an application. Recent developments of these high-performing mass spectrometers are mentioned to provide a future outlook. PMID:21742802

  8. Fractals and Transformations.

    ERIC Educational Resources Information Center

    Bannon, Thomas J.

    1991-01-01

    Discussed are several different transformations based on the generation of fractals including self-similar designs, the chaos game, the koch curve, and the Sierpinski Triangle. Three computer programs which illustrate these concepts are provided. (CW)

  9. Proof in Transformation Geometry

    ERIC Educational Resources Information Center

    Bell, A. W.

    1971-01-01

    The first of three articles showing how inductively-obtained results in transformation geometry may be organized into a deductive system. This article discusses two approaches to enlargement (dilatation), one using coordinates and the other using synthetic methods. (MM)

  10. Nontensorial Transformation Optics

    NASA Astrophysics Data System (ADS)

    García-Meca, C.; Barceló, C.

    2016-06-01

    We present an alternative version of transformation optics that allows us to mold the flow of light without rotating or scaling the electromagnetic fields. The resulting media experience unusual force densities, are nonreciprocal, and exhibit loss or gain. Because of these singular features, a variety of effects and devices unreachable by standard transformation optics can be achieved, including reflectionless light compression, optical modes with arbitrary in-plane polarization, and special isolators.

  11. Series Transmission Line Transformer

    DOEpatents

    Buckles, Robert A.; Booth, Rex; Yen, Boris T.

    2004-06-29

    A series transmission line transformer is set forth which includes two or more of impedance matched sets of at least two transmissions lines such as shielded cables, connected in parallel at one end ans series at the other in a cascading fashion. The cables are wound about a magnetic core. The series transmission line transformer (STLT) which can provide for higher impedance ratios and bandwidths, which is scalable, and which is of simpler design and construction.

  12. 25. VIEW, LOOKING SOUTHWEST INSIDE TRANSFORMER ROOM, SHOWING TRANSFORMERS AND ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    25. VIEW, LOOKING SOUTHWEST INSIDE TRANSFORMER ROOM, SHOWING TRANSFORMERS AND KNIFE SWITCHES - Sacramento River Bridge, Spanning Sacramento River at California State Highway 275, Sacramento, Sacramento County, CA

  13. Floral Transformation of Wheat

    NASA Astrophysics Data System (ADS)

    Agarwal, Sujata; Loar, Star; Steber, Camille; Zale, Janice

    A method is described for the floral transformation of wheat using a protocol similar to the floral dip of Arabidopsis. This method does not employ tissue culture of dissected embryos, but instead pre-anthesis spikes with clipped florets at the early, mid to late uninucleate microspore stage are dipped in Agrobacterium infiltration media harboring a vector carrying anthocyanin reporters and the NPTII selectable marker. T1 seeds are examined for color changes induced in the embryo by the anthocyanin reporters. Putatively transformed seeds are germinated and the seedlings are screened for the presence of the NPTII gene based on resistance to paromomycin spray and assayed with NPTII ELISAs. Genomic DNA of putative transformants is digested and analyzed on Southern blots for copy number to determine whether the T-DNA has integrated into the nucleus and to show the number of insertions. The non-optimized transformation efficiencies range from 0.3 to 0.6% (number of transformants/number of florets dipped) but the efficiencies are higher in terms of the number of transformants produced/number of seeds set ranging from 0.9 to 10%. Research is underway to maximize seed set and optimize the protocol by testing different Agrobacterium strains, visual reporters, vectors, and surfactants.

  14. Stable Crack Growth During Thermal Actuation of Shape Memory Alloys

    NASA Astrophysics Data System (ADS)

    Jape, S.; Baxevanis, T.; Lagoudas, D. C.

    2016-03-01

    A finite element analysis of crack growth is carried out in shape memory alloys subjected to thermal variations under plane strain, mode I, constant applied loading. The crack is assumed to propagate at a critical level of the crack-tip energy release rate which is modeled using the virtual crack closure technique. The load level, applied at a high temperature at which the austenite phase is stable, is assumed sufficiently low so that the resulting crack-tip energy release rate is smaller than the critical value but sufficiently high so that the critical value is reached during cooling, initiating crack growth (Baxevanis and Lagoudas in Int J Fract 191:191-213, 2015). Stable crack growth is observed, mainly associated with the shielding effect of the transformed material left in the wake of the advancing crack. Results pertaining to the near-tip mechanical fields and fracture toughness are presented and their sensitivity to phase transformation metrics and bias load levels is investigated.

  15. Stable zymomonas mobilis xylose and arabinose fermenting strains

    DOEpatents

    Zhang, Min; Chou, Yat-Chen

    2008-04-08

    The present invention briefly includes a transposon for stable insertion of foreign genes into a bacterial genome, comprising at least one operon having structural genes encoding enzymes selected from the group consisting of xylAxylB, araBAD and tal/tkt, and at least one promoter for expression of the structural genes in the bacterium, a pair of inverted insertion sequences, the operons contained inside the insertion sequences, and a transposase gene located outside of the insertion sequences. A plasmid shuttle vector for transformation of foreign genes into a bacterial genome, comprising at least one operon having structural genes encoding enzymes selected from the group consisting of xylAxylB, araBAD and tal/tkt, at least one promoter for expression of the structural genes in the bacterium, and at least two DNA fragments having homology with a gene in the bacterial genome to be transformed, is also provided.The transposon and shuttle vectors are useful in constructing significantly different Zymomonas mobilis strains, according to the present invention, which are useful in the conversion of the cellulose derived pentose sugars into fuels and chemicals, using traditional fermentation technology, because they are stable for expression in a non-selection medium.

  16. Glass transition and stable glass formation of tetrachloromethane.

    PubMed

    Chua, Y Z; Tylinski, M; Tatsumi, S; Ediger, M D; Schick, C

    2016-06-28

    Physical vapor deposition (PVD) has been used to prepare organic glasses with very high kinetic stability and it has been suggested that molecular anisotropy is a prerequisite for stable glass formation. Here we use PVD to prepare glasses of tetrachloromethane, a simple organic molecule with a nearly isotropic molecular structure. In situ AC nanocalorimetry was used to characterize the vapor-deposited glasses. Glasses of high kinetic stability were produced by deposition near 0.8 Tg. The isothermal transformation of the vapor-deposited glasses into the supercooled liquid state gave further evidence that tetrachloromethane forms glasses with high kinetic stability, with the transformation time exceeding the structural relaxation time of the supercooled liquid by a factor of 10(3). The glass transition temperature of liquid-cooled tetrachloromethane is determined as Tg ≈ 78 K, which is different from previously reported values. The frequency dependence of the glass transition was also determined and the fragility was estimated as m ≈ 118. The successful formation of PVD glasses of tetrachloromethane which have high kinetic stability argues that molecular asymmetry is not a prerequisite for stable glass formation. PMID:27369523

  17. Glass transition and stable glass formation of tetrachloromethane

    NASA Astrophysics Data System (ADS)

    Chua, Y. Z.; Tylinski, M.; Tatsumi, S.; Ediger, M. D.; Schick, C.

    2016-06-01

    Physical vapor deposition (PVD) has been used to prepare organic glasses with very high kinetic stability and it has been suggested that molecular anisotropy is a prerequisite for stable glass formation. Here we use PVD to prepare glasses of tetrachloromethane, a simple organic molecule with a nearly isotropic molecular structure. In situ AC nanocalorimetry was used to characterize the vapor-deposited glasses. Glasses of high kinetic stability were produced by deposition near 0.8 Tg. The isothermal transformation of the vapor-deposited glasses into the supercooled liquid state gave further evidence that tetrachloromethane forms glasses with high kinetic stability, with the transformation time exceeding the structural relaxation time of the supercooled liquid by a factor of 103. The glass transition temperature of liquid-cooled tetrachloromethane is determined as Tg ≈ 78 K, which is different from previously reported values. The frequency dependence of the glass transition was also determined and the fragility was estimated as m ≈ 118. The successful formation of PVD glasses of tetrachloromethane which have high kinetic stability argues that molecular asymmetry is not a prerequisite for stable glass formation.

  18. Relativistic Transformation of Solid Angle.

    ERIC Educational Resources Information Center

    McKinley, John M.

    1980-01-01

    Rederives the relativistic transformations of light intensity from compact sources (stars) to show where and how the transformation of a solid angle contributes. Discusses astrophysical and other applications of the transformations. (Author/CS)

  19. High Frequency Stable Oscillate boiling

    NASA Astrophysics Data System (ADS)

    Li, Fenfang; Gonzalez-Avila, Silvestre Roberto; Ohl, Claus Dieter

    2015-11-01

    We present an unexpected regime of resonant bubble oscillations on a thin metal film submerged in water, which is continuously heated with a focused CW laser. The oscillatory bubble dynamics reveals a remarkably stable frequency of several 100 kHz and is resolved from the side using video recordings at 1 million frames per second. The emitted sound is measured simultaneously and shows higher harmonics. Once the laser is switched on the water in contact with the metal layer is superheated and an explosively expanding cavitation bubble is generated. However, after the collapse a microbubble is nucleated from the bubble remains which displays long lasting oscillations. Generally, pinch-off from of the upper part of the microbubble is observed generating a continuous stream of small gas bubbles rising upwards. The cavitation expansion, collapse, and the jetting of gas bubbles are detected by the hydrophone and are correlated to the high speed video. We find the bubble oscillation frequency is dependent on the bubble size and surface tension. A preliminary model based on Marangoni flow and heat transfer can explain the high flow velocities observed, yet the origin of bubble oscillation is currently not well understood.

  20. Evolutionary routes to stable ownership.

    PubMed

    Hare, D; Reeve, H K; Blossey, B

    2016-06-01

    Ownership can evolve in potentially any species. Drawing on insights from across disciplines, we distinguish between possession and ownership and present species-neutral criteria for ownership, defined as respect for possession. We use a variant of the tug-of-war evolutionary game to demonstrate how ownership can evolve in the form of a new, biologically realistic strategy, Restraint With Retaliation (RWR). In our game, resource holding potential (RHP) is assumed to be equal between interactants, and resource holding asymmetry determines whether ownership is adaptive. RWR will be evolutionarily stable when the ratio of resource holdings between interactants is relatively low, but not when this ratio is sufficiently high. We offer RWR as one evolutionary route to ownership among many, and discuss how ownership unites previously described behavioural phenomena across taxa. We propose that some but not all mechanisms of territory formation and maintenance can be considered ownership, and show that territories are not the only resources that can be owned. We argue that ownership can be a powerful cooperative solution to tragedies of the commons and problems of collective action throughout the biological world. We advance recent scholarship that has begun to investigate the biological importance of ownership, and we call for a comprehensive account of its evolutionary logic and taxonomic distribution. We propose that ownership should be considered a fundamental, unifying biological phenomenon. PMID:26991035

  1. Photonic plasmid stability of transformed Salmonella typhimurium: A comparison of three unique plasmids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Acquiring a highly stable photonic plasmid in transformed Salmonella typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella typhimurium (S. typh-lux) u...

  2. Photonic Plasmid Stability of Transformed Salmonella Typhimurium: A Comparison of Three Unique Plasmids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Acquiring a highly stable photonic plasmid in transformed Salmonella Typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella Typhimurium (S....

  3. Advanced thermally stable jet fuels

    SciTech Connect

    Schobert, H.H.

    1999-01-31

    The Pennsylvania State University program in advanced thermally stable coal-based jet fuels has five broad objectives: (1) Development of mechanisms of degradation and solids formation; (2) Quantitative measurement of growth of sub-micrometer and micrometer-sized particles suspended in fuels during thermal stressing; (3) Characterization of carbonaceous deposits by various instrumental and microscopic methods; (4) Elucidation of the role of additives in retarding the formation of carbonaceous solids; (5) Assessment of the potential of production of high yields of cycloalkanes by direct liquefaction of coal. Future high-Mach aircraft will place severe thermal demands on jet fuels, requiring the development of novel, hybrid fuel mixtures capable of withstanding temperatures in the range of 400--500 C. In the new aircraft, jet fuel will serve as both an energy source and a heat sink for cooling the airframe, engine, and system components. The ultimate development of such advanced fuels requires a thorough understanding of the thermal decomposition behavior of jet fuels under supercritical conditions. Considering that jet fuels consist of hundreds of compounds, this task must begin with a study of the thermal degradation behavior of select model compounds under supercritical conditions. The research performed by The Pennsylvania State University was focused on five major tasks that reflect the objectives stated above: Task 1: Investigation of the Quantitative Degradation of Fuels; Task 2: Investigation of Incipient Deposition; Task 3: Characterization of Solid Gums, Sediments, and Carbonaceous Deposits; Task 4: Coal-Based Fuel Stabilization Studies; and Task 5: Exploratory Studies on the Direct Conversion of Coal to High Quality Jet Fuels. The major findings of each of these tasks are presented in this executive summary. A description of the sub-tasks performed under each of these tasks and the findings of those studies are provided in the remainder of this volume

  4. Leucine metabolism in stable cirrhosis.

    PubMed

    Mullen, K D; Denne, S C; McCullough, A J; Savin, S M; Bruno, D; Tavill, A S; Kalhan, S C

    1986-01-01

    Alterations in protein and amino acid metabolism have been postulated to explain the frequent observations of muscle wasting and decreased plasma branched-chain amino acid concentrations in cirrhosis. In order to investigate the changes in protein metabolism, we have measured the rates of leucine turnover and oxidation in six stable, biopsy-proven cirrhotics and six age and sex-matched healthy control subjects after an overnight fast, using [1-13C]leucine tracer. Following a primed constant-rate infusion of [1-13C]leucine, the 13C enrichments of plasma leucine and expired CO2 were used to estimate leucine turnover and oxidation, respectively. Fat-free body mass was estimated from the measurements of total body water as quantified by H2[18O] tracer dilution. The rates of CO2 production and oxygen consumption were measured hourly during the study period, using open-circuit respiratory calorimetry. Urinary urea, ammonia and total nitrogen excretion rates were quantified from timed urine samples. Even though the plasma leucine levels were lower in cirrhotics as compared with controls (100.5 +/- 17.1 vs. 138.3 +/- 20.4 mumoles per liter, mean +/- S.D., p less than 0.001), the rates of leucine turnover were not significantly different in the two groups (89.4 +/- 19.0 vs. 87.8 +/- 19.0 mumoles per kg X hr). In contrast, the rates of leucine oxidation were significantly reduced in cirrhosis (8.1 +/- 2.5 vs. 12.7 +/- 3.1 mumoles per kg X hr, p less than 0.01). When all subjects were considered, the leucine oxidation rate was correlated with plasma leucine concentration (r = 0.62, p less than 0.03).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3089896

  5. Transformation of the bioherbicide Colletotrichum gloeosporioides f. sp. Aeschynomene By electroporation of germinated conidia.

    PubMed

    Robinson, M; Sharon, A

    1999-08-01

    A highly efficient and reproducible procedure for the transformation of the bioherbicide Colletotrichum gloeosporioides f. sp. aeschynomene by electroporation of germinated conidia is reported. Development of the procedure involved a detailed study of the germination process followed by extensive calibration of the electrical-pulse and selection conditions. Optimization of the transformation protocol was facilitated by the use of the green fluorescent protein that helped in the identification of stable transformants and in a fast assessment of transgene expression levels, colony homogeneity and genetic stability. Under optimal conditions, over 80 stable transformants/cuvette were obtained. Plasmid integration was predominantly homologous, but high transformation rates were obtained both with homologous and non-homologous vectors. The method described not only opens up opportunities for the genetic manipulation of C. gloeosporioides f. sp. aeschynomene, but also provides a framework for the development and optimization of transformation in other fungi. PMID:10447601

  6. Martensitic transformation in zirconia

    SciTech Connect

    Deville, Sylvain . E-mail: sylvain.deville@insa-lyon.fr; Guenin, Gerard; Chevalier, Jerome

    2004-11-08

    We investigate by atomic force microscopy (AFM) the surface relief resulting from martensitic tetragonal to monoclinic phase transformation induced by low temperature autoclave aging in ceria-stabilized zirconia. AFM appears as a very powerful tool to investigate martensite relief quantitatively and with a great precision. The crystallographic phenomenological theory is used to predict the expected relief induced by the transformation, for the particular case of lattice correspondence ABC1, where tetragonal c axis becomes the monoclinic c axis. A model for variants spatial arrangement for this lattice correspondence is proposed and validated by the experimental observations. An excellent agreement is found between the quantitative calculations outputs and the experimental measurements at nanometer scale yielded by AFM. All the observed features are explained fully quantitatively by the calculations, with discrepancies between calculations and quantitative experimental measurements within the measurements and calculations precision range. In particular, the crystallographic orientation of the transformed grains is determined from the local characteristics of transformation induced relief. It is finally demonstrated that the strain energy is the controlling factor of the surface transformation induced by low temperature autoclave treatments in this material.

  7. On Nonstable and Stable Population Momentum

    PubMed Central

    Olgiati, Analia S.; Levin, Simon A.

    2014-01-01

    This article decomposes total population momentum into two constituent and multiplicative parts: “nonstable” momentum and “stable” momentum. Nonstable momentum depends on deviations between a population’s current age distribution and its implied stable age distribution. Stable momentum is a function of deviations between a population’s implied stable and stationary age distributions. In general, the factorization of total momentum into the product of nonstable and stable momentum is a very good approximation. The factorization is exact, however, when the current age distribution is stable or when observed fertility is already at replacement. We provide numerical illustrations by calculating nonstable, stable, and total momentum for 176 countries, the world, and its major regions. In short, the article brings together disparate strands of the population momentum literature and shows how the various kinds of momentum fit together into a single unifying framework. PMID:21948106

  8. Genetic manipulation of gamma-linolenic acid (GLA) synthesis in a commercial variety of evening primrose (Oenothera sp.).

    PubMed

    de Gyves, Emilio Mendoza; Sparks, Caroline A; Sayanova, Olga; Lazzeri, Paul; Napier, Johnathan A; Jones, Huw D

    2004-07-01

    A robust Agrobacterium-mediated transformation procedure was developed for Rigel, a commercial cultivar of evening primrose, and used to deliver a cDNA encoding a Delta(6)-desaturase from borage under the control of a cauliflower mosaic virus (CaMV) 35S promoter. Analysis of the transformed plants demonstrated an altered profile of polyunsaturated fatty acids, with an increase in gamma-linolenic acid and octadecatetraenoic acid in leaf tissues when compared with control lines. PMID:17134396

  9. Pineapple [Ananas comosus (L.) Merr].

    PubMed

    Gangopadhyay, Gaurab; Mukherjee, Kalyan K

    2015-01-01

    The efficacy of Agrobacterium-mediated pineapple transformation technique has been improved (mean percentage of transgenic micro-shoots regenerated from initial callus explants up to 20.6%) using a novel encapsulation-based, antibiotic selection procedure. The detailed protocol using a standard plant transformation vector (pCAMBIA1304) as reported in an 'elite' Indian variety (Queen) of pineapple [Ananas comosus (L.) Merr] can be applied to other varieties of pineapple for introgression of target genes. PMID:25416266

  10. Potato (Solanum tuberosum L.).

    PubMed

    Chetty, Venkateswari J; Narváez-Vásquez, Javier; Orozco-Cárdenas, Martha L

    2015-01-01

    Agrobacterium-mediated transformation is the most common method for the incorporation of foreign genes into the genome of potato as well as many other species in the Solanaceae family. This chapter describes protocols for the genetic transformation of three species of potato: Solanum tuberosum subsp. tuberosum (Desiréé), S. tuberosum subsp. andigenum (Blue potato), and S. tuberosum subsp. andigena using internodal segments as explants. PMID:25416251

  11. On spinors transformations

    NASA Astrophysics Data System (ADS)

    Budinich, Marco

    2016-07-01

    We begin showing that for even dimensional vector spaces V all automorphisms of their Clifford algebras are inner. So all orthogonal transformations of V are restrictions to V of inner automorphisms of the algebra. Thus under orthogonal transformations P and T—space and time reversal—all algebra elements, including vectors v and spinors φ, transform as v → xvx-1 and φ → xφx-1 for some algebra element x. We show that while under combined PT spinor φ → xφx-1 remains in its spinor space, under P or T separately φ goes to a different spinor space and may have opposite chirality. We conclude with a preliminary characterization of inner automorphisms with respect to their property to change, or not, spinor spaces.

  12. Translating and Transforming Care

    PubMed Central

    Gillespie, Alex; Moore, Helen

    2015-01-01

    This article examines how the Disability Living Allowance claim form, used in the United Kingdom to allocate £13 billion of disability benefits, translates and transforms disability and care. Twenty-two people with acquired brain injury and their main informal caregivers (n = 44) were video-recorded filling in the disability claim form. Participants disagreed on 26% of the questions, revealing two types of problems. Translation problems arose as participants struggled to provide categorical responses to ambiguous questions and were unable to report contextual variability in care needs or divergences of perception. Transformation problems arose as participants resisted the way in which the form positioned them, forcing them to conceptualize their relationship in terms of dependency and burden. The disability claim form co-opts claimants to translate care and disability into bureaucratically predefined categories, and it transforms the care relationship that it purports to document. PMID:25792487

  13. Transformational plasmon optics.

    PubMed

    Liu, Yongmin; Zentgraf, Thomas; Bartal, Guy; Zhang, Xiang

    2010-06-01

    We propose and demonstrate efficiently molding surface plasmon polaritons (SPPs) based on transformation optics. SPPs are surface modes of electromagnetic waves tightly bound at metal-dielectric interfaces, which allow us to scale optics beyond the diffraction limit. Taking advantage of transformation optics, here we show that the propagation of SPPs can be manipulated in a prescribed manner by careful control of the dielectric material properties adjacent to a metal. Since the metal properties are completely unaltered, this methodology provides a practical way for routing light at very small scales. For instance, our approach enables SPPs to travel at uneven and curved surfaces over a broad wavelength range, where SPPs would normally suffer significant scattering losses. In addition, a plasmonic 180 degrees waveguide bend and a plasmonic Luneburg lens with simple designs are presented. The unique design flexibility of the transformational plasmon optics introduced here may open a new door to nano optics and downscaling of photonic circuits. PMID:20465268

  14. Transformation based endorsement systems

    NASA Technical Reports Server (NTRS)

    Sudkamp, Thomas

    1988-01-01

    Evidential reasoning techniques classically represent support for a hypothesis by a numeric value or an evidential interval. The combination of support is performed by an arithmetic rule which often requires restrictions to be placed on the set of possibilities. These assumptions usually require the hypotheses to be exhausitive and mutually exclusive. Endorsement based classification systems represent support for the alternatives symbolically rather than numerically. A framework for constructing endorsement systems is presented in which transformations are defined to generate and update the knowledge base. The interaction of the knowledge base and transformations produces a non-monotonic reasoning system. Two endorsement based reasoning systems are presented to demonstrate the flexibility of the transformational approach for reasoning with ambiguous and inconsistent information.

  15. Ultraviolet and thermally stable polymer compositions

    NASA Technical Reports Server (NTRS)

    Adamson, M. J.; Gloria, H. R.; Goldsberry, R. E.; Reinisch, R. F.

    1972-01-01

    Copolymers, produced from aromatic substituted aromatic azine-siloxane compositions, are thermally stable, solar ultraviolet light non-degradable by wavelengths shorter than those reaching earth surface.

  16. Stable molecular configuration in crystalline carboxylic acids

    NASA Astrophysics Data System (ADS)

    Hayashi, Soichi; Umemura, Junzo; Nakamura, Ryoko

    1980-12-01

    The stable (lower enthalpy) molecular configurations of propionic, butyric, Jeric and lauric acids in the crystalline state have been examined via their atom-atom potentials. It was found that the cis configuration is more stable than the trans configuration for propionic, butyric and valeric acids, and that the trans configuration is more stable than the cis configuration for lauric acid, in accord with a previous IR spectral analysis. The potential energy of benzoic acid was recalculated using the positions of atoms given by Speakman, and indicates that the A form is more stable than the B form, in agreement with the results of previous work.

  17. Germ-line transformation of the Queensland fruit fly, Bactrocera tryoni, using a piggyBac vector in the presence of endogenous piggyBac elements

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report the stable genetic transformation of the Queensland fruit fly Bactrocera tryoni using a piggyBac vector marked with either the fluorescent protein DsRed or EGFP.A transformation frequency of 5–10% was obtained.Inheritance of the transgenes has remained stable over eight generations despite...

  18. Plastid transformation in soybean.

    PubMed

    Dubald, Manuel; Tissot, Ghislaine; Pelissier, Bernard

    2014-01-01

    The biotechnological potential of plastid genetic engineering has been illustrated in a limited number of higher plant species. We have developed a reproducible method to generate plastid transformants in soybean (Glycine max), a crop of major agronomic importance. The transformation vectors are delivered to embryogenic cultures by the particle gun method and selection performed using the aadA antibiotic resistance gene. Homoplasmy is established rapidly in the selected events without the need for further selection or regeneration cycles, and genes of interest can be expressed at a high level in green tissues. This is a significant step toward the commercial application of this technology. PMID:24599865

  19. Successful Transformational Radiology Leaders.

    PubMed

    Douget, Karen

    2015-01-01

    Transformational radiology leaders elevate subordinates, expand self-awareness, develop lasting relationships, strive to exceed expectations, and uphold the vision and goals of the organization. In order for radiology leaders to become more transformational in their leadership style there are four fundamental elements they must learn: idealized influence, individualized consideration, inspirational motivation, and intellectual stimulation. Leaders can utilize personality and self-assessments to learn more about themselves, identify areas of strengths and weaknesses, and learn to be more effective when leading employees. PMID:26710553

  20. Development of Stable Solidification Method for Insoluble Ferrocyanides-13170

    SciTech Connect

    Ikarashi, Yuki; Masud, Rana Syed; Mimura, Hitoshi; Ishizaki, Eiji; Matsukura, Minoru

    2013-07-01

    The development of stable solidification method of insoluble ferrocyanides sludge is an important subject for the safety decontamination in Fukushima NPP-1. By using the excellent immobilizing properties of zeolites such as gas trapping ability and self-sintering properties, the stable solidification of insoluble ferrocyanides was accomplished. The immobilization ratio of Cs for K{sub 2}[CoFe(CN){sub 6}].nH{sub 2}O saturated with Cs{sup +} ions (Cs{sub 2}[CoFe(CN){sub 6}].nH{sub 2}O) was estimated to be less than 0.1% above 1,000 deg. C; the adsorbed Cs{sup +} ions are completely volatilized. In contrast, the novel stable solid form was produced by the press-sintering of the mixture of Cs{sub 2}[CoFe(CN){sub 6}].nH{sub 2}O and zeolites at higher temperature of 1,000 deg. C and 1,100 deg. C; Cs volatilization and cyanide release were completely depressed. The immobilization ratio of Cs, under the mixing conditions of Cs{sub 2}[CoFe(CN){sub 6}].nH{sub 2}O:CP= 1:1 and calcining temperature: 1,000 deg. C, was estimated to be nearly 100%. As for the kinds of zeolites, natural mordenite (NM), clinoptilolite (CP) and Chabazite tended to have higher immobilization ratio compared to zeolite A. This may be due to the difference in the phase transformation between natural zeolites and synthetic zeolite A. In the case of the composites (K{sub 2-X}Ni{sub X/2}[NiFe(CN){sub 6}].nH{sub 2}O loaded natural mordenite), relatively high immobilization ratio of Cs was also obtained. This method using zeolite matrices can be applied to the stable solidification of the solid wastes of insoluble ferrocyanides sludge. (authors)

  1. Finger millet [Eleusine coracana (L.) Gaertn].

    PubMed

    Ceasar, Stanislaus Antony; Ignacimuthu, Savarimuthu

    2015-01-01

    Millets are the primary food source for millions of people in tropical regions of the world supplying mineral nutrition and protein. In this chapter, we describe an optimized protocol for the Agrobacterium-mediated transformation of finger millet variety GPU 45. Agrobacterium strain LBA4404 harboring plasmid pCAMBIA1301 which contains hygromycin phosphotransferase (hph) as selectable marker gene and β-glucuronidase (GUS) as reporter gene has been used. This protocol utilizes the shoot apex explants for the somatic embryogenesis and regeneration of finger millet after the transformation by Agrobacterium. Desiccation of explants during cocultivation helps for the better recovery of transgenic plants. This protocol is very useful for the efficient production of transgenic plants in finger millet through Agrobacterium-mediated transformation. PMID:25300836

  2. Focus On: Transforming Relationships.

    ERIC Educational Resources Information Center

    Karal, Pearl; And Others

    1985-01-01

    Karal describes the psychodynamics by which children learn to fear and distrust long-term commitments. Vaines proposes a conceptualization of the home economist as transforming actor. LeBow addresses methods of treating obese youth. Fewster and Kuhonta offer insights on communicating with Third World rural women. (SK)

  3. Transforming Data into Knowledge

    ERIC Educational Resources Information Center

    Mills, Lane

    2006-01-01

    School systems can be data rich and information poor if they do not understand and manage their data effectively. The task for school leaders is to put existing data into a format that lends itself to answering questions and improving outcomes for the students. Common barriers to transforming data into knowledge in education settings often include…

  4. Transformative Mixed Methods Research

    ERIC Educational Resources Information Center

    Mertens, Donna M.

    2010-01-01

    Paradigms serve as metaphysical frameworks that guide researchers in the identification and clarification of their beliefs with regard to ethics, reality, knowledge, and methodology. The transformative paradigm is explained and illustrated as a framework for researchers who place a priority on social justice and the furtherance of human rights.…

  5. Fourier Transform Mass Spectrometry.

    ERIC Educational Resources Information Center

    Gross, Michael L.; Rempel, Don L.

    1984-01-01

    Discusses the nature of Fourier transform mass spectrometry and its unique combination of high mass resolution, high upper mass limit, and multichannel advantage. Examines its operation, capabilities and limitations, applications (ion storage, ion manipulation, ion chemistry), and future applications and developments. (JN)

  6. Transforming Primary Mathematics

    ERIC Educational Resources Information Center

    Askew, Mike

    2011-01-01

    What is good mathematics teaching? What is mathematics teaching good for? Who is mathematics teaching for? These are just some of the questions addressed in "Transforming Primary Mathematics", a highly timely new resource for teachers which accessibly sets out the key theories and latest research in primary maths today. Under-pinned by findings…

  7. Transforming Young Lives

    ERIC Educational Resources Information Center

    Larson, Scott

    2014-01-01

    Discussions of transformational change pervade the field of business but are rare in work with young people at risk--those most in need of deep change. Instead, the nation seems preoccupied with punishing or medicating problem behavior. Some propose the alternative of "rehabilitation," but that term means "to restore to former…

  8. Computerized toroidal transformer design

    NASA Technical Reports Server (NTRS)

    1970-01-01

    Computer program designs transformers which have one primary /center tap permissible/ and up to 20 untapped secondaries, and which can handle up to 500 V across any one winding. Computer determines total secondary power, core type, primary turns, secondary turns, and wire sizes for primary and secondary windings.

  9. Fixture for winding transformers

    NASA Technical Reports Server (NTRS)

    Mclyman, M. T.

    1980-01-01

    Bench-mounted fixture assists operator in winding toroid-shaped transformer cores. Toroid is rigidly held in place as wires are looped around. Arrangement frees both hands for rapid winding and untangling of wires that occurs when core is hand held.

  10. Transformer and Meter Tester

    NASA Technical Reports Server (NTRS)

    Stoms, R. M.

    1984-01-01

    Numerically-controlled 5-axis machine tool uses transformer and meter to determine and indicate whether tool is in home position, but lacks built-in test mode to check them. Tester makes possible test, and repair of components at machine rather then replace them when operation seems suspect.

  11. The fast Hartley transform

    NASA Astrophysics Data System (ADS)

    Mar, Mark H.

    1990-11-01

    The purpose of this paper is to report the results of testing the fast Hartley transform (FHT) and comparing it with the fast Fourier transform (FFT). All the definitions and equations in this paper are quoted and cited from the series of references. The author of this report developed a FORTRAN program which computes the Hartley transform. He tested the program with a generalized electromagnetic pulse waveform and verified the results with the known value. Fourier analysis is an essential tool to obtain frequency domain information from transient time domain signals. The FFT is a popular tool to process many of today's audio and electromagnetic signals. System frequency response, digital filtering of signals, and signal power spectrum are the most practical applications of the FFT. However, the Fourier integral transform of the FFT requires computer resources appropriate for the complex arithmetic operations. On the other hand, the FHT can accomplish the same results faster and requires fewer computer resources. The FHT is twice as fast as the FFT, uses only half the computer resources, and so could be more useful than the FFT in typical applications such as spectral analysis, signal processing, and convolution. This paper presents a FORTRAN computer program for the FHT algorithm along with a brief description and compares the results and performance of the FHT and the FFT algorithms.

  12. Teachers Transform Lives.

    ERIC Educational Resources Information Center

    Bradshaw, Delia

    2001-01-01

    Teachers transform lives, and the ripple effect goes on for years. Three pertinent questions are asked in this paper: Where does this power come from? What is its source? and What makes teachers so special? Two aspects of these questions are the multiplicity of identities that coexist within each teacher and the passion inside teachers that…

  13. Transformation Problem Solving Abilities.

    ERIC Educational Resources Information Center

    Harmel, Sarah Jane

    The relationship between transformation problem performance and Guilford Structure of Intellect (SI) abilities is explored. During two group sessions 42 females and 35 males, age 18-39, were administered 12 Guilford SI tests exemplifying all five symbolic content (numeric) operations, and three contents in the divergent production area. Logical…

  14. Leading System Transformation

    ERIC Educational Resources Information Center

    Harris, Alma

    2010-01-01

    This article outlines progress made towards education transformation in Wales. It explores the way in which tri-level reform is guiding system level change in Wales and looks at the implementation of the School Effectiveness Framework, which is at the heart of the reform process. It describes the way in which professional learning communities are…

  15. Dual Christoffel Transformations

    NASA Astrophysics Data System (ADS)

    Odake, S.; Sasaki, R.

    2011-07-01

    Crum's theorem and its modification à la Krein-Adler are formulated for the discrete quantum mechanics with real shifts, whose eigenfunctions consist of orthogonal polynomials of a discrete variable. The modification produces the associated polynomials with a finite number of degrees deleted. This in turn provides the well known Christoffel transformation for the dual orthogonal polynomials with the corresponding positions deleted.

  16. Global Transformations and Agriculture.

    ERIC Educational Resources Information Center

    Campbell, Rex R.

    1990-01-01

    Examines worldwide political, economic, and social transformations and their impact on agriculture, focusing on biotechnology. Discusses rise of international corporations and accompanying constraints on government power. Sees trend toward increasing agribusiness role in world food and agricultural sectors. Calls for broader views and research in…

  17. Winthrop College Transformed Curriculum.

    ERIC Educational Resources Information Center

    Hawisher, Margaret F.

    Dealing with the issue of a changing society and recognizing that teacher education has remained basically unchanged for 100 years, the faculty of the Winthrop College School of Education agreed to take the risk involved with transforming the teacher education curriculum. Three interdisciplinary teams have identified curriculum to be taught to…

  18. Transforming Education with Technology

    ERIC Educational Resources Information Center

    Scherer, Marge

    2011-01-01

    In this EL interview, Karen Cator, the director of the Office of Educational Technology at the U.S. Department of Education, talks about ways to realize the potential of technology to transform education. She discusses what students need: their own digital devices for classroom use, the ability to use the information they access, the skills to…

  19. Transformer Impedance Reflection Demonstration

    ERIC Educational Resources Information Center

    Layton, William

    2014-01-01

    Questions often arise as to how a device attached to a transformer can draw power from the electrical power grid since it seems that the primary and secondary are not connected to one another. However, a closer look at how the primary and secondary are linked together magnetically and a consideration of the role of Lenz's law in this linkage…

  20. Beyond Reform: Transformation

    ERIC Educational Resources Information Center

    Davidson, Jill

    2007-01-01

    The Coalition of Essential Schools (CES) is not a reform movement. To reform is to make a thing again; reformation implies a stasis that doesn't deliver enough for the educational future. This issue of Horace demonstrates that Essential schools and the districts and networks that support them are at various points in the journey of transformation,…

  1. Improved Transformation of Anthurium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methods to increase transformation efficiency and yields of transgenic Anthurium andraeanum Linden ex. André hybrids were sought while effecting gene transfer for resistance to the two most important pests, bacterial blight (Xanthomonas axonopodis pv. dieffenbachiae) and nematodes (Radopholus simili...

  2. Transformative Change Initiative

    ERIC Educational Resources Information Center

    Bragg, D. D.; Kirby, C.; Witt, M. A.; Richie, D.; Mix, S.; Feldbaum, M.; Liu, S.; Mason, M.

    2014-01-01

    The Transformative Change Initiative (TCI) is dedicated to assisting community colleges to scale up innovation in the form of guided pathways, programs of study, and evidence-based strategies to improve student outcomes and program, organization, and system performance. The impetus for TCI is the Trade Adjustment Assistance Community College and…

  3. Transforming American Education

    ERIC Educational Resources Information Center

    Horn, Michael B.; Mackey, Katherine

    2011-01-01

    In this article the authors accept as a given the National Education Technology Plan's vision of a transformed education system powered by technology such that learners receive personalized and engaging learning experiences, and where assessment, teaching, infrastructure, and productivity are redefined. The article analyzes this vision of a…

  4. Inflated Soft Actuators with Reversible Stable Deformations.

    PubMed

    Hines, Lindsey; Petersen, Kirstin; Sitti, Metin

    2016-05-01

    Most soft robotic systems are currently dependent on bulky compressors or pumps. A soft actuation method is presented combining hyperelastic membranes and dielectric elastomer actuators to switch between stable deformations of sealed chambers. This method is capable of large repeatable deformations, and has a number of stable states proportional to the number of actuatable membranes in the chamber. PMID:27008455

  5. Blood feeding behavior of the stable fly

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Stable fly is a fly that looks similar to a house fly but both sexes are blood feeders. Blood is required for successful fertilization and development of eggs. Bites are painful but there is usually no pain after the fly stops feeding. The stable fly is a persistent feeder and will continue trying t...

  6. Rainbow Fourier Transform

    NASA Technical Reports Server (NTRS)

    Alexandrov, Mikhail D.; Cairns, Brian; Mishchenko, Michael I.

    2012-01-01

    We present a novel technique for remote sensing of cloud droplet size distributions. Polarized reflectances in the scattering angle range between 135deg and 165deg exhibit a sharply defined rainbow structure, the shape of which is determined mostly by single scattering properties of cloud particles, and therefore, can be modeled using the Mie theory. Fitting the observed rainbow with such a model (computed for a parameterized family of particle size distributions) has been used for cloud droplet size retrievals. We discovered that the relationship between the rainbow structures and the corresponding particle size distributions is deeper than it had been commonly understood. In fact, the Mie theory-derived polarized reflectance as a function of reduced scattering angle (in the rainbow angular range) and the (monodisperse) particle radius appears to be a proxy to a kernel of an integral transform (similar to the sine Fourier transform on the positive semi-axis). This approach, called the rainbow Fourier transform (RFT), allows us to accurately retrieve the shape of the droplet size distribution by the application of the corresponding inverse transform to the observed polarized rainbow. While the basis functions of the proxy-transform are not exactly orthogonal in the finite angular range, this procedure needs to be complemented by a simple regression technique, which removes the retrieval artifacts. This non-parametric approach does not require any a priori knowledge of the droplet size distribution functional shape and is computationally fast (no look-up tables, no fitting, computations are the same as for the forward modeling).

  7. A stable bovine papillomavirus hybrid plasmid that expresses a dominant selective trait.

    PubMed Central

    Law, M F; Byrne, J C; Howley, P M

    1983-01-01

    We describe a bovine papillomavirus hybrid plasmid containing the neomycin resistance gene from Tn5 inserted into a mammalian cell transcriptional unit. This plasmid is maintained as a stable extrachromosomal element (20 to 100 copies per diploid genome) in mouse cells selected either for the transformed phenotype or for resistance to the aminoglycoside G418. Cells selected for G418 resistance initially display a flat, nontransformed phenotype before exhibiting the gross morphological characteristics of transformation. The delay in the appearance of the transformed phenotype indicated that some intracellular event or series of events subsequent to the establishment of transcriptionally active bovine papillomavirus 1 hybrid plasmid is required for the manifestation of the transformed phenotype. Images PMID:6318082

  8. Construction of energy-stable projection-based reduced order models

    SciTech Connect

    Kalashnikova, Irina; Barone, Matthew F.; Arunajatesan, Srinivasan; van Bloemen Waanders, Bart G.

    2014-12-15

    Our paper aims to unify and extend several approaches for building stable projection-based reduced order models (ROMs) using the energy method and the concept of “energy-stability”. Attention is focused on linear time-invariant (LTI) systems. First, an approach for building energy stable Galerkin ROMs for linear hyperbolic or incompletely parabolic systems of partial differential equations (PDEs) using continuous projection is proposed. The key idea is to apply to the system a transformation induced by the Lyapunov function for the system, and to build the ROM in the transformed variables. The result of this procedure will be a ROM that is energy-stable for any choice of reduced basis. It is shown that, for many PDE systems, the desired transformation is induced by a special inner product, termed the “symmetry inner product”. Next, attention is turned to building energy-stable ROMs via discrete projection. A discrete counterpart of the continuous symmetry inner product, termed the “Lyapunov inner product”, is derived. Moreover, it is shown that the Lyapunov inner product can be computed in a black-box fashion for a stable LTI system ari sing from the discretization of a system of PDEs in space. Projection in this inner product guarantees a ROM that is energy-stable, again for any choice of reduced basis. Connections between the Lyapunov inner product and the inner product induced by the balanced truncation algorithm are made. We also made comparisons between the symmetry inner product and the Lyapunov inner product. Performance of ROMs constructed using these inner products is evaluated on several benchmark test cases.

  9. Cotton (Gossypium hirsutum L.).

    PubMed

    Rathore, Keerti S; Campbell, LeAnne M; Sherwood, Shanna; Nunes, Eugenia

    2015-01-01

    Cotton continues to be a crop of great economic importance in many developing and some developed countries. Cotton plants expressing the Bt gene to deter some of the major pests have been enthusiastically and widely accepted by the farmers in three of the major producing countries, i.e., China, India, and the USA. Considering the constraints related to its production and the wide variety of products derived from the cotton plant, it offers several target traits that can be improved through genetic engineering. Thus, there is a great need to accelerate the application of biotechnological tools for cotton improvement. This requires a simple, yet robust gene delivery/transformant recovery system. Recently, a protocol, involving large-scale, mechanical isolation of embryonic axes from germinating cottonseeds followed by direct transformation of the meristematic cells has been developed by an industrial laboratory. However, complexity of the mechanical device and the patent restrictions are likely to keep this method out of reach of most academic laboratories. In this chapter, we describe the method developed in our laboratory that has undergone further refinements and involves Agrobacterium-mediated transformation of cotton cells, selection of stable transgenic callus lines, and recovery of plants via somatic embryogenesis. PMID:25416245

  10. Lead transformation to pyromorphite by fungi.

    PubMed

    Rhee, Young Joon; Hillier, Stephen; Gadd, Geoffrey Michael

    2012-02-01

    Lead (Pb) is a serious environmental pollutant in all its chemical forms [1]. Attempts have been made to immobilize lead in soil as the mineral pyromorphite using phosphate amendments (e.g., rock phosphate, phosphoric acid, and apatite [2-5]), although our work has demonstrated that soil fungi are able to transform pyromorphite into lead oxalate [6, 7]. Lead metal, an important structural and industrial material, is subject to weathering, and soil contamination also occurs through hunting and shooting [8, 9]. Although fungi are increasingly appreciated as geologic agents [10-12], there is a distinct lack of knowledge about their involvement in lead geochemistry. We examined the influence of fungal activity on lead metal and discovered that metallic lead can be transformed into chloropyromorphite, the most stable lead mineral that exists. This is of geochemical significance, not only regarding lead fate and cycling in the environment but also in relation to the phosphate cycle and linked with microbial transformations of inorganic and organic phosphorus. This paper provides the first report of mycogenic chloropyromorphite formation from metallic lead and highlights the significance of this phenomenon as a biotic component of lead biogeochemistry, with additional consequences for microbial survival in lead-contaminated environments and bioremedial treatments for Pb-contaminated land. PMID:22245002

  11. Reduction/Transformation Operators

    SciTech Connect

    Bartlett, Roscoe A.

    2006-09-01

    RTOp (reduction/transformation operators) is a collection of C++ software that provides the basic mechanism for implementinig vector operations in a flexible and efficient manner. This is the main interface utilized by Thyra to allow for the specification of specific vector reduction and/or transformation operations. The RTOp package contains three different types of software. (a) a small number of interoperability interfaces. (b) support software including code for the parallel SPMD mode based on only Teuchos::Comm(and not MPl directly(, and (c) a library of pre-implemented RTOp subclasses for everything from simple AXPYs and norms, to more specialized vector operations. RTOp allows an algorithm developer to implement their own RTOp subclasses in a way that is independent from any specific serial, parallel, out-of-core or other type of vector implementation. RTOp is a required package by Thyra and MOOCHO. (c)

  12. Transforming the optical landscape.

    PubMed

    Pendry, J B; Luo, Yu; Zhao, Rongkuo

    2015-05-01

    Electromagnetism provides us with some of the most powerful tools in science, encompassing lasers, optical microscopes, magnetic resonance imaging scanners, radar, and a host of other techniques. To understand and develop the technology requires more than a set of formal equations. Scientists and engineers have to form a vivid picture that fires their imaginations and enables intuition to play a full role in the process of invention. It is to this end that transformation optics has been developed, exploiting Faraday's picture of electric and magnetic fields as lines of force, which can be manipulated by the electrical permittivity and magnetic permeability of surrounding materials. Transformation optics says what has to be done to place the lines of force where we want them to be. PMID:25931549

  13. Dynamically tunable transformation thermodynamics

    NASA Astrophysics Data System (ADS)

    García-Meca, Carlos; Barceló, Carlos

    2016-04-01

    Recently, the introduction of transformation thermodynamics has provided a way to design thermal media that alter the flow of heat according to any spatial deformation, enabling the construction of novel devices such as thermal cloaks or concentrators. However, in its current version, this technique only allows static deformations of space. Here, we develop a space–time theory of transformation thermodynamics that incorporates the possibility of performing time-varying deformations. This extra freedom greatly widens the range of achievable effects, providing an additional degree of control for heat management applications. As an example, we design a reconfigurable thermal cloak that can be opened and closed dynamically, therefore being able to gradually adjust the temperature distribution of a given region.

  14. High voltage isolation transformer

    NASA Technical Reports Server (NTRS)

    Clatterbuck, C. H.; Ruitberg, A. P. (Inventor)

    1985-01-01

    A high voltage isolation transformer is provided with primary and secondary coils separated by discrete electrostatic shields from the surfaces of insulating spools on which the coils are wound. The electrostatic shields are formed by coatings of a compound with a low electrical conductivity which completely encase the coils and adhere to the surfaces of the insulating spools adjacent to the coils. Coatings of the compound also line axial bores of the spools, thereby forming electrostatic shields separating the spools from legs of a ferromagnetic core extending through the bores. The transformer is able to isolate a high constant potential applied to one of its coils, without the occurrence of sparking or corona, by coupling the coatings, lining the axial bores to the ferromagnetic core and by coupling one terminal of each coil to the respective coating encasing the coil.

  15. Transformation of Bacillus subtilis.

    PubMed

    Zhang, Xiao-Zhou; You, Chun; Zhang, Yi-Heng Percival

    2014-01-01

    Bacillus subtilis has tremendous applications in both academic research and industrial production. However, molecular cloning and transformation of B. subtilis are not as easy as those of Escherichia coli. Here we developed a simple protocol based on super-competent cells prepared from the recombinant B. subtilis strain SCK6 and multimeric plasmids generated by prolonged overlap extension-PCR. Super-competent B. subtilis SCK6 cells were prepared by overexpression of the competence master regulator ComK that was induced by adding xylose. This new protocol is simple (e.g., restriction enzyme, phosphatase, and ligase free), fast, and highly efficient (i.e., ~10(7) or ~10(4) transformants per μg of multimeric plasmid or ligated plasmid DNA, respectively). Shuttle vectors for E. coli-B. subtilis are not required. PMID:24838881

  16. A DC Transformer

    NASA Technical Reports Server (NTRS)

    Youngquist, Robert C.; Ihlefeld, Curtis M.; Starr, Stanley O.

    2013-01-01

    A component level dc transformer is described in which no alternating currents or voltages are present. It operates by combining features of a homopolar motor and a homopolar generator, both de devices, such that the output voltage of a de power supply can be stepped up (or down) with a corresponding step down (or up) in current. The basic theory for this device is developed, performance predictions are made, and the results from a small prototype are presented. Based on demonstrated technology in the literature, this de transformer should be scalable to low megawatt levels, but it is more suited to high current than high voltage applications. Significant development would be required before it could achieve the kilovolt levels needed for de power transmission.

  17. Transforming the optical landscape

    NASA Astrophysics Data System (ADS)

    Pendry, J. B.; Luo, Yu; Zhao, Rongkuo

    2015-05-01

    Electromagnetism provides us with some of the most powerful tools in science, encompassing lasers, optical microscopes, magnetic resonance imaging scanners, radar, and a host of other techniques. To understand and develop the technology requires more than a set of formal equations. Scientists and engineers have to form a vivid picture that fires their imaginations and enables intuition to play a full role in the process of invention. It is to this end that transformation optics has been developed, exploiting Faraday’s picture of electric and magnetic fields as lines of force, which can be manipulated by the electrical permittivity and magnetic permeability of surrounding materials. Transformation optics says what has to be done to place the lines of force where we want them to be.

  18. Reduction/Transformation Operators

    2006-09-01

    RTOp (reduction/transformation operators) is a collection of C++ software that provides the basic mechanism for implementinig vector operations in a flexible and efficient manner. This is the main interface utilized by Thyra to allow for the specification of specific vector reduction and/or transformation operations. The RTOp package contains three different types of software. (a) a small number of interoperability interfaces. (b) support software including code for the parallel SPMD mode based on only Teuchos::Comm(and notmore » MPl directly(, and (c) a library of pre-implemented RTOp subclasses for everything from simple AXPYs and norms, to more specialized vector operations. RTOp allows an algorithm developer to implement their own RTOp subclasses in a way that is independent from any specific serial, parallel, out-of-core or other type of vector implementation. RTOp is a required package by Thyra and MOOCHO. (c)« less

  19. Bacterial transformation of terpenoids

    NASA Astrophysics Data System (ADS)

    Grishko, V. V.; Nogovitsina, Y. M.; Ivshina, I. B.

    2014-04-01

    Data on the bacterial transformation of terpenoids published in the literature in the past decade are analyzed. Possible pathways for chemo-, regio- and stereoselective modifications of terpenoids are discussed. Considerable attention is given to new technological approaches to the synthesis of terpenoid derivatives suitable for the use in the perfume and food industry and promising as drugs and chiral intermediates for fine organic synthesis. The bibliography includes 246 references.

  20. Transformer Impedance Reflection Demonstration

    NASA Astrophysics Data System (ADS)

    Layton, William

    2014-10-01

    Questions often arise as to how a device attached to a transformer can draw power from the electrical power grid since it seems that the primary and secondary are not connected to one another. However, a closer look at how the primary and secondary are linked together magnetically and a consideration of the role of Lenz's law in this linkage may help to clear up this confusion.

  1. Transforming revenue management.

    PubMed

    Silveria, Richard; Alliegro, Debra; Nudd, Steven

    2008-11-01

    Healthcare organizations that want to undertake a patient administrative/revenue management transformation should: Define the vision with underlying business objectives and key performance measures. Strategically partner with key vendors for business process development and technology design. Create a program organization and governance infrastructure. Develop a corporate design model that defines the standards for operationalizing the vision. Execute the vision through technology deployment and corporate design model implementation. PMID:18990839

  2. Laminated piezoelectric transformer

    NASA Technical Reports Server (NTRS)

    Vazquez Carazo, Alfredo (Inventor)

    2006-01-01

    A laminated piezoelectric transformer is provided using the longitudinal vibration modes for step-up voltage conversion applications. The input portions are polarized to deform in a longitudinal plane and are bonded to an output portion. The deformation of the input portions is mechanically coupled to the output portion, which deforms in the same longitudinal direction relative to the input portion. The output portion is polarized in the thickness direction relative its electrodes, and piezoelectrically generates a stepped-up output voltage.

  3. Absence of a stable atomic structure in fluorinated graphene.

    PubMed

    Boukhvalov, Danil W

    2016-05-21

    Based on the results of first-principles calculations we demonstrate that significant distortion of graphene sheets caused by adsorption of fluorine atoms leads to the formation of metastable patterns for which the next step of fluorination is considerably less energetically favorable. Existence of these stable patterns oriented along the armchair direction makes possible the synthesis of various CFx structures. The combination of strong distortion of the nonfluorinated graphene sheet with the doping caused by the polar nature of C-F bonds reduces the energy cost of migration and the energy of migration barriers, making possible the migration of fluorine atoms on the graphene surface as well as transformation of the shapes of fluorinated areas. The decreasing energy cost of migration with increasing fluorine content also leads to increasing numbers of single fluorine adatoms, which could be the source of magnetic moments. PMID:27116897

  4. Function transformation without reinforcement.

    PubMed

    Tonneau, Franćois; Arreola, Fara; Martínez, Alma Gabriela

    2006-05-01

    In studies of function transformation, participants initially are taught to match stimuli in the presence of a contextual cue, X; the stimuli to be matched bear some formal relation to each other, for example, a relation of opposition or difference. In a second phase, the participants are taught to match arbitrary stimuli (say, A and B) in the presence of X. In a final test, A often displays behavioral functions that differ from those of B, and can be predicted from the nature of the relation associated with X in the initial training phase. Here we report function-transformation effects in the absence of selection responses and of their reinforcers. In three experiments with college students, exposure to relations of difference or identity modified the responses given to later stimuli. In Experiment 1, responses to a test stimulus A varied depending on preexposure to pairs of colors that were distinct from A but exemplified relations of difference or identity. In Experiment 2, a stimulus A acquired distinct functions, depending on its previous pairing with a contextual cue X that had itself been paired with identity or difference among colors. Experiment 3 confirmed the results of Experiment 2 with a modified design. Our data are consistent with the notion that relations of identity or difference can serve as stimuli for Pavlovian processes, and, in compound with other cues, produce apparent function-transformation effects. PMID:16776058

  5. Function Transformation without Reinforcement

    PubMed Central

    Tonneau, François; Arreola, Fara; Martínez, Alma Gabriela

    2006-01-01

    In studies of function transformation, participants initially are taught to match stimuli in the presence of a contextual cue, X; the stimuli to be matched bear some formal relation to each other, for example, a relation of opposition or difference. In a second phase, the participants are taught to match arbitrary stimuli (say, A and B) in the presence of X. In a final test, A often displays behavioral functions that differ from those of B, and can be predicted from the nature of the relation associated with X in the initial training phase. Here we report function-transformation effects in the absence of selection responses and of their reinforcers. In three experiments with college students, exposure to relations of difference or identity modified the responses given to later stimuli. In Experiment 1, responses to a test stimulus A varied depending on preexposure to pairs of colors that were distinct from A but exemplified relations of difference or identity. In Experiment 2, a stimulus A acquired distinct functions, depending on its previous pairing with a contextual cue X that had itself been paired with identity or difference among colors. Experiment 3 confirmed the results of Experiment 2 with a modified design. Our data are consistent with the notion that relations of identity or difference can serve as stimuli for Pavlovian processes, and, in compound with other cues, produce apparent function-transformation effects. PMID:16776058

  6. Transforming the dilemma

    PubMed Central

    Taylor, Christine; Nowak, Martin A.

    2008-01-01

    How does natural selection lead to cooperation between competing individuals? The Prisoner's Dilemma captures the essence of this problem. Two players can either cooperate or defect. The payoff for mutual cooperation, R, is greater than the payoff for mutual defection, P. But a defector versus a cooperator receives the highest payoff, T, while the cooperator obtains the lowest payoff, S. Hence, the Prisoner's Dilemma is defined by the payoff ranking T > R > P > S. In a well-mixed population, defectors always have a higher expected payoff than cooperators, and therefore natural selection favors defectors. The evolution of cooperation requires specific mechanisms. Here we discuss five mechanisms for the evolution of cooperation: direct reciprocity, indirect reciprocity, kin selection, group selection and network reciprocity (or graph selection). Each mechanism leads to a transformation of the Prisoner's Dilemma payoff matrix. From the transformed matrices, we derive the fundamental conditions for the evolution of cooperation. The transformed matrices can be used in standard frameworks of evolutionary dynamics such as the replicator equation or stochastic processes of game dynamics in finite populations. PMID:17711471

  7. Origin and Fate of Organic Compounds in Water: Characterization by Compound-Specific Stable Isotope Analysis

    NASA Astrophysics Data System (ADS)

    Schmidt, Torsten C.; Jochmann, Maik A.

    2012-07-01

    Within the past 15 years, compound-specific stable isotope analysis has continued to increase in popularity in the area of contaminant hydrology of organic molecules. In particular, in cases where concentration data alone are insufficient to elucidate environmental processes unequivocally, the isotope signature can provide additional unique information. Specifically, it can help answer questions about contaminant source apportionment, quantification of biotic and abiotic processes, and identification of transformation reactions on a mechanistic level. We review advances in laboratory and field investigations and exemplary applications in contaminant hydrology via stable isotope analysis. We also highlight future directions in the field.

  8. Bi-stable optical element actuator device

    DOEpatents

    Holdener, Fred R.; Boyd, Robert D.

    2002-01-01

    The present invention is a bistable optical element actuator device utilizing a powered means to move an actuation arm, to which an optical element is attached, between two stable positions. A non-powered means holds the actuation arm in either of the two stable positions. The optical element may be a electromagnetic (EM) radiation or particle source, an instrument, or EM radiation or particle transmissive reflective or absorptive elements. A bearing is used to transfer motion and smoothly transition the actuation arm between the two stable positions.

  9. 60. VIEW OF THE CURRENT TRANSFORMER VAULT. THIS CURRENT TRANSFORMER ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    60. VIEW OF THE CURRENT TRANSFORMER VAULT. THIS CURRENT TRANSFORMER WAS USED TO SENSE HIGH CURRENT BEING GENERATED ON GENERATOR NUMBER 3 AND REDUCE IT TO A LOWER, EXACT ANALOG VALUE THAT COULD BE SAFELY HANDLED AND MONITORED WITH THE CONTROL CIRCUITRY. THE CURRENT TRANSFORMER IS LOCATED IN THE CENTER OF THE PHOTOGRAPH. THE CONNECTING BUS ABOVE THE TRANSFORMER WAS REMOVED FOR SALVAGE. - New York, New Haven & Hartford Railroad, Cos Cob Power Plant, Sound Shore Drive, Greenwich, Fairfield County, CT

  10. Predicting unit performance by assessing transformational and transactional leadership.

    PubMed

    Bass, Bernard M; Avolio, Bruce J; Jung, Dong I; Berson, Yair

    2003-04-01

    How do leadership ratings collected from units operating under stable conditions predict subsequent performance of those units operating under high stress and uncertainty? To examine this question, the authors calculated the predictive relationships for the transformational and transactional leadership of 72 light infantry rifle platoon leaders for ratings of unit potency, cohesion, and performance for U.S. Army platoons participating in combat simulation exercises. Both transformational and transactional contingent reward leadership ratings of platoon leaders and sergeants positively predicted unit performance. The relationship of platoon leadership to performance was partially mediated through the unit's level of potency and cohesion. Implications, limitations, and future directions for leadership research are discussed. PMID:12731705

  11. 134. VIEW OF TRANSFORMER CABINETS ON NORTH SIDE OF TRANSFORMER ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    134. VIEW OF TRANSFORMER CABINETS ON NORTH SIDE OF TRANSFORMER ROOM (212), LSB (BLDG. 751), FACING SOUTH. POWER PANEL B AT EAST SIDE OF TRANSFORMER ROOM (212), FACING WEST, AT RIGHT IN PHOTOGRAPH. - Vandenberg Air Force Base, Space Launch Complex 3, Launch Pad 3 East, Napa & Alden Roads, Lompoc, Santa Barbara County, CA

  12. DNA transformations of Candida tropicalis with replicating and integrative vectors.

    PubMed

    Sanglard, D; Fiechter, A

    1992-12-01

    The alkane-assimilating yeast Candida tropicalis was used as a host for DNA transformations. A stable ade2 mutant (Ha900) obtained by UV-mutagenesis was used as a recipient for different vectors carrying selectable markers. A first vector, pMK16, that was developed for the transformation of C. albicans and carries an ADE2 gene marker and a Candida autonomously replicating sequence (CARS) element promoting autonomous replication, was compatible for transforming Ha900. Two transformant types were observed: (i) pink transformants which easily lose pMK16 under non-selective growth conditions; (ii) white transformants, in which the same plasmid exhibited a higher mitotic stability. In both cases pMK16 could be rescued from these cells in Escherichia coli. A second vector, pADE2, containing the isolated C. tropicalis ADE2, gene, was used to transform Ha900. This vector integrated in the yeast genome at homologous sites of the ade2 locus. Different integration types were observed at one or both ade2 alleles in single or in tandem repeats. PMID:1293885

  13. Alfalfa (Medicago sativa L.).

    PubMed

    Fu, Chunxiang; Hernandez, Timothy; Zhou, Chuanen; Wang, Zeng-Yu

    2015-01-01

    Alfalfa (Medicago sativa L.) is a high-quality forage crop widely grown throughout the world. This chapter describes an efficient protocol that allows for the generation of large number of transgenic alfalfa plants by sonication-assisted Agrobacterium-mediated transformation. Binary vectors carrying different selectable marker genes that confer resistance to phosphinothricin (bar), kanamycin (npt II), or hygromycin (hph) were used to generate transgenic alfalfa plants. Intact trifoliates collected from clonally propagated plants in the greenhouse were sterilized with bleach and then inoculated with Agrobacterium strain EHA105. More than 80 % of infected leaf pieces could produce rooted transgenic plants in 4-5 months after Agrobacterium-mediated transformation. PMID:25300843

  14. Applications of stable isotopes in clinical pharmacology

    PubMed Central

    Schellekens, Reinout C A; Stellaard, Frans; Woerdenbag, Herman J; Frijlink, Henderik W; Kosterink, Jos G W

    2011-01-01

    This review aims to present an overview of the application of stable isotope technology in clinical pharmacology. Three main categories of stable isotope technology can be distinguished in clinical pharmacology. Firstly, it is applied in the assessment of drug pharmacology to determine the pharmacokinetic profile or mode of action of a drug substance. Secondly, stable isotopes may be used for the assessment of drug products or drug delivery systems by determination of parameters such as the bioavailability or the release profile. Thirdly, patients may be assessed in relation to patient-specific drug treatment; this concept is often called personalized medicine. In this article, the application of stable isotope technology in the aforementioned three areas is reviewed, with emphasis on developments over the past 25 years. The applications are illustrated with examples from clinical studies in humans. PMID:21801197

  15. Stable Isotope Signatures for Microbial Forensics

    SciTech Connect

    Kreuzer, Helen W.

    2012-01-03

    The isotopic distribution of the atoms composing the molecules of microorganisms is a function of the substrates used by the organisms. The stable isotope content of an organism is fixed so long as no further substrate consumption and biosynthesis occurs, while the radioactive isotopic content decays over time. The distribution of stable isotopes of C, N, O and H in heterotrophic microorganisms is a direct function of the culture medium, and therefore the stable isotope composition can be used to associate samples with potential culture media and also with one another. The 14C content depends upon the 14C content, and therefore the age, of the organic components of the culture medium, as well as on the age of the culture itself. Stable isotope signatures can thus be used for sample matching, to associate cultures with specific growth media, and to predict characteristics of growth media.

  16. DNA modifications: Another stable base in DNA

    NASA Astrophysics Data System (ADS)

    Brazauskas, Pijus; Kriaucionis, Skirmantas

    2014-12-01

    Oxidation of 5-methylcytosine has been proposed to mediate active and passive DNA demethylation. Tracking the history of DNA modifications has now provided the first solid evidence that 5-hydroxymethylcytosine is a stable epigenetic modification.

  17. Thermally Stable Piezoelectric and Pyroelectric Polymers

    NASA Technical Reports Server (NTRS)

    Simpson, Joycelyn O.; St. Clair, Terry L.

    2006-01-01

    A class of thermally stable piezoelectric and pyroelectric polymers, and an improved method of making them, have been invented. These polymers can be used as substrates for a wide variety of electromechanical transducers, sensors, and actuators.

  18. Computing Stable Outcomes in Hedonic Games

    NASA Astrophysics Data System (ADS)

    Gairing, Martin; Savani, Rahul

    We study the computational complexity of finding stable outcomes in symmetric additively-separable hedonic games. These coalition formation games are specified by an undirected edge-weighted graph: nodes are players, an outcome of the game is a partition of the nodes into coalitions, and the utility of a node is the sum of incident edge weights in the same coalition. We consider several natural stability requirements defined in the economics literature. For all of them the existence of a stable outcome is guaranteed by a potential function argument, so local improvements will converge to a stable outcome and all these problems are in PLS. The different stability requirements correspond to different local search neighbourhoods. For different neighbourhood structures, our findings comprise positive results in the form of polynomial-time algorithms for finding stable outcomes, and negative (PLS-completeness) results.

  19. Evolutionary origin of asymptotically stable consensus.

    PubMed

    Tang, Chang-Bing; Wu, Bin; Wang, Jian-Bo; Li, Xiang

    2014-01-01

    Consensus is widely observed in nature as well as in society. Up to now, many works have focused on what kind of (and how) isolated single structures lead to consensus, while the dynamics of consensus in interdependent populations remains unclear, although interactive structures are everywhere. For such consensus in interdependent populations, we refer that the fraction of population adopting a specified strategy is the same across different interactive structures. A two-strategy game as a conflict is adopted to explore how natural selection affects the consensus in such interdependent populations. It is shown that when selection is absent, all the consensus states are stable, but none are evolutionarily stable. In other words, the final consensus state can go back and forth from one to another. When selection is present, there is only a small number of stable consensus state which are evolutionarily stable. Our study highlights the importance of evolution on stabilizing consensus in interdependent populations. PMID:24699444

  20. Polymeric foams stable at high temperatures

    NASA Technical Reports Server (NTRS)

    Riccitiello, S. R.; Harrison, E. S.; Delano, C. B.

    1976-01-01

    Crosslinked poly(N-arylenebenzimidazoles) are stable up to 370 C. Polymers are made by mixing appropriate stoichiometric amounts of tetramine and aromatic dicarboxylic acid anhydride with phenol or alkyl-substituted phenol.

  1. Suppression of the chemically transformed phenotype of BHK cells by a human cDNA

    SciTech Connect

    Eiden, M.V.; MacArthur, L.; Okayama, Hiroto )

    1991-10-01

    Transformation of baby hamster kidney cell line BHK SN-10 by chemical carcinogens such as nitrosylmethylurea (NMU) is mediated by the loss of a gene product critical for the suppression of malignant transformation. Somatic cell hybrids between chemically transformed BHK SN-10 cells and either normal hamster kidney or human fibroblast cells are nontransformed; therefore, a recessive mechanism underlies the malignant transformation of BHK SN-10 cells after chemical carcinogenesis. A human fibroblast cDNA library was constructed and introduced into NMU-transformed BHK SN-10 cells (NMU 34m) in order to identify a human cDNA capable of suppressing cellular transformation. NMU-transformed BHK cells were analyzed for reversion to an anchorage-dependent stable reversion of NMU 34m cells encodes the intermediate filament protein vimentin, which is apparently required for maintenance of the normal phenotype in BHK SN-10 cells.

  2. RECENT ADVANCES IN BARLEY TRANSFORMATION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley, an important member of the cereals, has been successfully transformed through various methods such as particle bombardment, Agrobacterium-tumefaciens, DNA uptake, and electroporation. Initially, the transformation in barley concentrated on developing protocols using marker genes such as gus,...

  3. The Transformation of Disabilities Organizations

    ERIC Educational Resources Information Center

    Schalock, Robert L.; Verdugo, Miguel-Angel

    2013-01-01

    This article summarizes the five major characteristics of the transformation era and describes how intellectual and closely related developmental disabilities organizations can apply specific transformation strategies associated with each characteristic. Collectively, the characteristics and strategies provide a framework for transformation…

  4. Transformational Leadership, Integrity, and Power

    ERIC Educational Resources Information Center

    Harrison, Laura M.

    2011-01-01

    Transformational leadership enjoys widespread appeal among student affairs professionals. National Association of Student Personnel Administrators (NASPA) and American College Personnel Association (ACPA) conferences frequently feature speakers who promote transformational leadership's two primary tenets: (1) change is the central purpose of…

  5. Stable vector bundles and string theory

    SciTech Connect

    Gomez, Tomas L.; Sols, Ignacio; Lukic, Sergio

    2009-05-06

    In [4], Braun, He, Ovrut and Pantev proposed a model of string theory (based on the Calabi-Yau 3-fold X) whose low energy limit predicts certain properties of the Standard Model of particle Physics. This model depends on two vector bundles that have to be stable. We calculate the ample cone of X, and prove that one of them is stable, and the other one is not.

  6. Stable single-layer honeycomblike structure of silica.

    PubMed

    Özçelik, V Ongun; Cahangirov, S; Ciraci, S

    2014-06-20

    Silica or SiO(2), the main constituent of Earth's rocks has several 3D complex crystalline and amorphous phases, but it does not have a graphitelike layered structure in 3D. Our theoretical analysis and numerical calculations from the first principles predict a single-layer honeycomblike allotrope, hα silica, which can be viewed to be derived from the oxidation of silicene and it has intriguing atomic structure with reentrant bond angles in hexagons. It is a wide band gap semiconductor, which attains remarkable electromechanical properties showing geometrical changes under an external electric field. In particular, it is an auxetic metamaterial with a negative Poisson's ratio and has a high piezoelectric coefficient. While it can form stable bilayer and multilayer structures, its nanoribbons can show metallic or semiconducting behavior depending on their chirality. Coverage of dangling Si orbitals by foreign adatoms can attribute new functionalities to hα silica. In particular, Si(2)O(5), where Si atoms are saturated by oxygen atoms from top and bottom sides alternatingly can undergo a structural transformation to make silicatene, another stable, single layer structure of silica. PMID:24996101

  7. Stable Isotope Resolved Metabolomics Studies in Ex Vivo TIssue Slices

    PubMed Central

    Fan, Teresa W-M.; Lane, Andrew N.; Higashi, Richard M.

    2016-01-01

    An important component of this methodology is to assess the role of the tumor microenvironment on tumor growth and survival. To tackle this problem, we have adapted the original approach of Warburg 1, by combining thin tissue slices with Stable Isotope Resolved Metabolomics (SIRM) to determine detailed metabolic activity of human tissues. SIRM enables the tracing of metabolic transformations of source molecules such as glucose or glutamine over defined time periods, and is a requirement for detailed pathway tracing and flux analysis. In our approach, we maintain freshly resected tissue slices (both cancerous and non- cancerous from the same organ of the same subject) in cell culture media, and treat with appropriate stable isotope-enriched nutrients, e.g. 13C6-glucose or 13C5, 15N2 -glutamine. These slices are viable for at least 24 h, and make it possible to eliminate systemic influence on the target tissue metabolism while maintaining the original 3D cellular architecture. It is therefore an excellent pre-clinical platform for assessing the effect of therapeutic agents on target tissue metabolism and their therapeutic efficacy on individual patients 2,3. PMID:27158639

  8. Stable Single-Layer Honeycomblike Structure of Silica

    NASA Astrophysics Data System (ADS)

    Özçelik, V. Ongun; Cahangirov, S.; Ciraci, S.

    2014-06-01

    Silica or SiO2, the main constituent of Earth's rocks has several 3D complex crystalline and amorphous phases, but it does not have a graphitelike layered structure in 3D. Our theoretical analysis and numerical calculations from the first principles predict a single-layer honeycomblike allotrope, hα silica, which can be viewed to be derived from the oxidation of silicene and it has intriguing atomic structure with reentrant bond angles in hexagons. It is a wide band gap semiconductor, which attains remarkable electromechanical properties showing geometrical changes under an external electric field. In particular, it is an auxetic metamaterial with a negative Poisson's ratio and has a high piezoelectric coefficient. While it can form stable bilayer and multilayer structures, its nanoribbons can show metallic or semiconducting behavior depending on their chirality. Coverage of dangling Si orbitals by foreign adatoms can attribute new functionalities to hα silica. In particular, Si2O5, where Si atoms are saturated by oxygen atoms from top and bottom sides alternatingly can undergo a structural transformation to make silicatene, another stable, single layer structure of silica.

  9. Thin-Film Power Transformers

    NASA Technical Reports Server (NTRS)

    Katti, Romney R.

    1995-01-01

    Transformer core made of thin layers of insulating material interspersed with thin layers of ferromagnetic material. Flux-linking conductors made of thinner nonferromagnetic-conductor/insulator multilayers wrapped around core. Transformers have geometric features finer than those of transformers made in customary way by machining and mechanical pressing. In addition, some thin-film materials exhibit magnetic-flux-carrying capabilities superior to those of customary bulk transformer materials. Suitable for low-cost, high-yield mass production.

  10. Nanowire growth by an electron beam induced massive phase transformation

    SciTech Connect

    Sood, Shantanu; Kisslinger, Kim; Gouma, Perena

    2014-11-15

    Tungsten trioxide nanowires of a high aspect ratio have been synthesized in-situ in a TEM under an electron beam of current density 14A/cm² due to a massive polymorphic reaction. Sol-gel processed pseudocubic phase nanocrystals of tungsten trioxide were seen to rapidly transform to one dimensional monoclinic phase configurations, and this reaction was independent of the substrate on which the material was deposited. The mechanism of the self-catalyzed polymorphic transition and accompanying radical shape change is a typical characteristic of metastable to stable phase transformations in nanostructured polymorphic metal oxides. A heuristic model is used to confirm the metastable to stable growth mechanism. The findings are important to the control electron beam deposition of nanowires for functional applications starting from colloidal precursors.

  11. [Stable morphologies and mosaicism in the macroevolution of birds (Neornithes)].

    PubMed

    Zelenkov, N V

    2015-01-01

    Modern orders of Neornithine birds are separated by distinct morphological gaps, hampering the development of particular models. of macroevolutionary transformations. However, recent decades have witnessed the discovery and extensive study of many fossil groups of birds, which shed light on the origin of modern higher taxa. These fossils further allow analyzing the whole process of the macroevolutionary change in this group of vertebrates during the Cenozoic. Here, a brief review is presented of the "transitional" taxa with special attention to representative of the clade Galloanseres. Almost all transitional groups of fossil birds do display mosaic morphology. It is noted that many stable morphologies (characters and character complexes) persist in different groups, occurring either as primitive states, or as advanced ones. The stable recurrence of these characters is caused by the persistence of stable ontogenetic pathways. On the other hand, independent evolution of various morphologies can be explained by the modular organization of the development. The modularity in the structure of the foot is used here as an example to show a possible way of the origin of the foot in extant perching birds (Passeriformes). The mosaic morphology of the transitional taxa is seen as the only possible way to overcome the evolutionary constraint of the parallel optimization of numerous organismal systems. It is noted that in course of macroevolutionary change, no complete filling of morphological gaps occur. Transitional forms are separated by additional gaps from their ancestors and descendants. Hence, stem groups are seen as the most probable ancestors of crown taxa. It is further noted that the evolutionary formation of some higher taxa of the living birds was initiated by the evolution of advanced morphology of the feeding apparatus and skull, while change in their. postcranial morphology was delayed. PMID:26353395

  12. Agrobacterium tumefaciens-mediated transformation of the causative agent of Valsa canker of apple tree Valsa mali var. mali.

    PubMed

    Hu, Yang; Dai, Qingqing; Liu, Yangyang; Yang, Zhe; Song, Na; Gao, Xiaoning; Voegele, Ralf Thomas; Kang, Zhensheng; Huang, Lili

    2014-06-01

    Valsa mali var. mali (Vmm), which is the causative agent of Valsa canker of apple tree, causes heavy damage to apple production in eastern Asia. In this article, we report Agrobacterium tumefaciens-mediated transformation (ATMT) of Vmm and expression of gfp (green fluorescent protein) in this fungus. The transformation system was optimized to a transformation efficiency of approximately 150 transformants/10(6) conidia, and a library containing over 4,000 transformants was generated. The tested transformants were mitotically stable. One hundred percent hph (hygromycin B phosphotransferase) integration into Vmm was identified by PCR and five single-copy integration of T-DNA was detected in the eighteen transformants by Southern blot. To our knowledge, this is the first report of ATMT of Vmm. Furthermore, this library has been used to identify genes involved in the virulence of the pathogen, and the transformation system may also be useful to the transformation of other species of the genus Valsa. PMID:24554343

  13. Canonical Transformations of Kepler Trajectories

    ERIC Educational Resources Information Center

    Mostowski, Jan

    2010-01-01

    In this paper, canonical transformations generated by constants of motion in the case of the Kepler problem are discussed. It is shown that canonical transformations generated by angular momentum are rotations of the trajectory. Particular attention is paid to canonical transformations generated by the Runge-Lenz vector. It is shown that these…

  14. Enhancing Understanding of Transformation Matrices

    ERIC Educational Resources Information Center

    Dick, Jonathan; Childrey, Maria

    2012-01-01

    With the Common Core State Standards' emphasis on transformations, teachers need a variety of approaches to increase student understanding. Teaching matrix transformations by focusing on row vectors gives students tools to create matrices to perform transformations. This empowerment opens many doors: Students are able to create the matrices for…

  15. Teacher Transformation: Transcending Hegemonic Roots

    ERIC Educational Resources Information Center

    Peters, Gregory

    2012-01-01

    In this dissertation study, I build from the research on transformative teaching (transformative for students), liberating theories (Liberating, Liberation and Liberating Theories) as well as literature about transformation, reflection and discourse to make the case that our historic and continuously inequitable results for students based on…

  16. Quality as Transformation: Educational Metamorphosis

    ERIC Educational Resources Information Center

    Cheng, Ming

    2014-01-01

    The notion of "quality as transformation" has been widely used in the higher education sector. However, both quality and transformation are elusive terms. There is little research exploring how quality could be equated to transformation in the learning process. This paper will provide an insight into the relationship between quality and…

  17. The transformation of disabilities organizations.

    PubMed

    Schalock, Robert L; Verdugo, Miguel-Angel

    2013-08-01

    This article summarizes the five major characteristics of the transformation era and describes how intellectual and closely related developmental disabilities organizations can apply specific transformation strategies associated with each characteristic. Collectively, the characteristics and strategies provide a framework for transformation thinking, learning, and acting. Specific application examples are given. PMID:23909588

  18. Novel and potential application of cryopreservation to plant genetic transformation.

    PubMed

    Wang, Biao; Zhang, Zhibo; Yin, Zhenfang; Feng, Chaohong; Wang, Qiaochun

    2012-01-01

    The world population now is 6.7 billion and is predicted to reach 9 billion by 2050. Such a rapid growing population has tremendously increased the challenge for food security. Obviously, it is impossible for traditional agriculture to ensure the food security, while plant biotechnology offers considerable potential to realize this goal. Over the last 15 years, great benefits have been brought to sustainable agriculture by commercial cultivation of genetically modified (GM) crops. Further development of new GM crops will with no doubt contribute to meeting the requirements for food by the increasing population. The present article provides updated comprehensive information on novel and potential application of cryopreservation to genetic transformation. The major progresses that have been achieved in this subject include (1), long-term storage of a large number of valuable plant genes, which offers a good potential for further development of novel cultivars by genetic transformation; (2), retention of regenerative capacity of embryogenic tissues and protoplasts, which ensures efficient plant regeneration system for genetic transformation; (3), improvement of transformation efficiency and plant regeneration of transformed cells; (4), long-term preservation of transgenic materials with stable expression of transgenes and productive ability of recombinant proteins, which allows transgenic materials to be stored in a safe manner before being analyzed and evaluated, and allows establishment of stable seed stocks for commercial production of homologous proteins. Data provided in this article clearly demonstrate that cryo-technique has an important role to play in the whole chain of genetic transformation. Further studies coupling cryotechnique and genetic transformation are expected to significantly improve development of new GM crops. PMID:22079800

  19. Banded transformer cores

    NASA Technical Reports Server (NTRS)

    Mclyman, C. W. T. (Inventor)

    1974-01-01

    A banded transformer core formed by positioning a pair of mated, similar core halves on a supporting pedestal. The core halves are encircled with a strap, selectively applying tension whereby a compressive force is applied to the core edge for reducing the innate air gap. A dc magnetic field is employed in supporting the core halves during initial phases of the banding operation, while an ac magnetic field subsequently is employed for detecting dimension changes occurring in the air gaps as tension is applied to the strap.

  20. Thermal energy transformer

    NASA Technical Reports Server (NTRS)

    Berdahl, C. M.; Thiele, C. L. (Inventor)

    1979-01-01

    For use in combination with a heat engine, a thermal energy transformer is presented. It is comprised of a flux receiver having a first wall defining therein a radiation absorption cavity for converting solar flux to thermal energy, and a second wall defining an energy transfer wall for the heat engine. There is a heat pipe chamber interposed between the first and second walls having a working fluid disposed within the chamber and a wick lining the chamber for conducting the working fluid from the second wall to the first wall. Thermal energy is transferred from the radiation absorption cavity to the heat engine.

  1. Graph Transformation with Dependencies for the Specification of Interactive Systems

    NASA Astrophysics Data System (ADS)

    Corradini, Andrea; Foss, Luciana; Ribeiro, Leila

    We propose a notion of Graph Transformation Systems ( gts) with dependency relation, more expressive than a previously proposed one, and suitable for the specification of interactions. We show how a specification using gts with dependencies can be implemented, at a lower level of abstraction, by a transactional gts, that is, a gts equipped with the notion of observable (stable) items in which computations that correspond to “complete” interactions are characterized as transactions.

  2. Computer simulation of martensitic transformations

    SciTech Connect

    Xu, Ping

    1993-11-01

    The characteristics of martensitic transformations in solids are largely determined by the elastic strain that develops as martensite particles grow and interact. To study the development of microstructure, a finite-element computer simulation model was constructed to mimic the transformation process. The transformation is athermal and simulated at each incremental step by transforming the cell which maximizes the decrease in the free energy. To determine the free energy change, the elastic energy developed during martensite growth is calculated from the theory of linear elasticity for elastically homogeneous media, and updated as the transformation proceeds.

  3. Transformational silicon electronics.

    PubMed

    Rojas, Jhonathan Prieto; Torres Sevilla, Galo Andres; Ghoneim, Mohamed Tarek; Inayat, Salman Bin; Ahmed, Sally M; Hussain, Aftab Mustansir; Hussain, Muhammad Mustafa

    2014-02-25

    In today's traditional electronics such as in computers or in mobile phones, billions of high-performance, ultra-low-power devices are neatly integrated in extremely compact areas on rigid and brittle but low-cost bulk monocrystalline silicon (100) wafers. Ninety percent of global electronics are made up of silicon. Therefore, we have developed a generic low-cost regenerative batch fabrication process to transform such wafers full of devices into thin (5 μm), mechanically flexible, optically semitransparent silicon fabric with devices, then recycling the remaining wafer to generate multiple silicon fabric with chips and devices, ensuring low-cost and optimal utilization of the whole substrate. We show monocrystalline, amorphous, and polycrystalline silicon and silicon dioxide fabric, all from low-cost bulk silicon (100) wafers with the semiconductor industry's most advanced high-κ/metal gate stack based high-performance, ultra-low-power capacitors, field effect transistors, energy harvesters, and storage to emphasize the effectiveness and versatility of this process to transform traditional electronics into flexible and semitransparent ones for multipurpose applications. PMID:24476361

  4. Image Transformations-Montserrat

    NASA Technical Reports Server (NTRS)

    2002-01-01

    A slightly oblique digital photograph of Montserrat taken from the International Space Station was posted to Earth Observatory in December 2001. An Earth Observatory reader used widely available software to correct the oblique perspective and adjust the color. The story of how he modified the image includes step-by-step instructions that can be applied to other photographs. Photographs of Earth taken by astronauts have shaped our view of the Earth and are part of our popular culture because NASA makes them easily accessible to the public. Read the Transformations Story for more information. The original image was digital photograph number ISS002-E-9309, taken on July 9, 2001, from the International Space Station and was provided by the Earth Sciences and Image Analysis Laboratory at Johnson Space Center. Additional images taken by astronauts and cosmonauts can be viewed at the NASA-JSC Gateway to Astronaut Photography of Earth. Bill Innanen provided the transformed image and the story of how he did it.

  5. Construction of energy-stable projection-based reduced order models

    DOE PAGESBeta

    Kalashnikova, Irina; Barone, Matthew F.; Arunajatesan, Srinivasan; van Bloemen Waanders, Bart G.

    2014-12-15

    Our paper aims to unify and extend several approaches for building stable projection-based reduced order models (ROMs) using the energy method and the concept of “energy-stability”. Attention is focused on linear time-invariant (LTI) systems. First, an approach for building energy stable Galerkin ROMs for linear hyperbolic or incompletely parabolic systems of partial differential equations (PDEs) using continuous projection is proposed. The key idea is to apply to the system a transformation induced by the Lyapunov function for the system, and to build the ROM in the transformed variables. The result of this procedure will be a ROM that is energy-stablemore » for any choice of reduced basis. It is shown that, for many PDE systems, the desired transformation is induced by a special inner product, termed the “symmetry inner product”. Next, attention is turned to building energy-stable ROMs via discrete projection. A discrete counterpart of the continuous symmetry inner product, termed the “Lyapunov inner product”, is derived. Moreover, it is shown that the Lyapunov inner product can be computed in a black-box fashion for a stable LTI system ari sing from the discretization of a system of PDEs in space. Projection in this inner product guarantees a ROM that is energy-stable, again for any choice of reduced basis. Connections between the Lyapunov inner product and the inner product induced by the balanced truncation algorithm are made. We also made comparisons between the symmetry inner product and the Lyapunov inner product. Performance of ROMs constructed using these inner products is evaluated on several benchmark test cases.« less

  6. High-Frequency Transformation of Undeveloped Plastids in Tobacco Suspension Cells

    PubMed Central

    Langbecker, Camri L.; Ye, Guang-Ning; Broyles, Debra L.; Duggan, Lisa L.; Xu, Charles W.; Hajdukiewicz, Peter T.J.; Armstrong, Charles L.; Staub, Jeffrey M.

    2004-01-01

    Although leaf chloroplast transformation technology was developed more than a decade ago, no reports exist of stable transformation of undeveloped plastids or other specialized plastid types, such as proplastids, etioplasts, or amyloplasts. In this work we report development of a dark-grown tobacco suspension cell model system to investigate the transformation potential of undeveloped plastids. Electron microscope analysis confirmed that the suspension cells carry plastids that are significantly smaller (approximately 50-fold less in volume) and have a very different subcellular localization and developmental state than leaf cell chloroplasts. Using antibiotic selection in the light, we demonstrated that both plastid and nuclear transformation of these cell suspensions is efficient and reproducible, with plastid transformation frequency at least equal to that of leaf chloroplast transformation. Homoplasmic plastid transformants are readily obtained in cell colonies, or in regenerated plants, providing a more consistent and versatile model than the leaf transformation system. Because of the uniformity of the cell suspension model, we could further show that growth rate, selection scheme, particle size, and DNA amount influence the frequency of transformation. Our results indicate that the rate-limiting steps for nuclear and plastid transformation are different, and each must be optimized separately. The suspension cell system will be useful as a model for understanding transformation in those plant species that utilize dark-grown embryogenic cultures and for characterizing the steps that lead to homoplasmic plastid transformation. PMID:15141065

  7. Transformations of emotional experience.

    PubMed

    de Cortiñas, Lia Pistiner

    2013-06-01

    In this paper the author approaches mental pain and the problems in a psychoanalytic treatment of patients with difficulties in the psychic transformation of their emotional experiences. The author is interested in the symbolic failure related to the obstruction of development of phantasies, dreams, dream-thoughts, etc. She differentiates symbolization disturbances related to hypertrophic projective identification from a detention of these primitive communications and emotional isolation. She puts forward the conjecture that one factor in the arrest of this development is the detention of projective identifications and that, when this primitive means of communication is re-established in a container-contained relationship of mutual benefit, this initiates the development of a symbolization process that can replace the pathological 'protection'. Another hypothesis she develops is that of inaccessible caesuras that, associated with the detention of projective identification, obstruct any integrative or interactive movement. This caesura and the detention of projective identifications affect mental functions needed for dealing with mental pain. The personality is left with precarious mental equipment for transforming emotional experiences. How can a psychoanalytical process stimulate the development of creative symbolization, transforming the emotional experiences and leading towards mental growth? The author approaches the clinical problem with the metaphor of the psychic birth of emotional experience. The modulation of mental pain in a container-contained relationship is a central problem for the development of the human mind. For discovering and giving a meaning to emotional experience, the infant depends on reverie, a function necessary in order to develop an evolved consciousness capable of being aware, which is different from the rudimentary consciousness that perceives but does not understand. The development of mature mental equipment is associated with the

  8. Advanced stable lipid-based formulations for a patient-centric product design.

    PubMed

    Becker, Karin; Saurugger, Eva-Maria; Kienberger, Diana; Lopes, Diogo; Haack, Detlev; Köberle, Martin; Stehr, Michael; Lochmann, Dirk; Zimmer, Andreas; Salar-Behzadi, Sharareh

    2016-01-30

    Multiparticulate dosage forms are a recent strategy to meet the special needs of children, elderly people and patients suffering from dysphagia. Our study presents a novel and cost-efficient approach for the manufacturing of a taste-masked multiparticulate system with a stable immediate release profile by applying lipid-based excipients in a solvent-free hot melt coating process. The thermosensitive N-acetylcysteine (N-ac) was used as model drug and hot-melt coated with a mixture of tripalmitin and polysorbate 65. A predictive in vitro method for the evaluation of the taste masking efficiency was developed based on the deprotonation of the carboxyl group of N-ac and the decline of pH, responsible for the unpleasant sour taste of the compound. The method was confirmed using in vivo studies. Differential scanning calorimetry and X-ray scattering experiments revealed polymorphic transformation and its dependency on transformation time, temperature and emulsifier concentration. During the process, the coating was transformed almost completely into the stable β-polymorph, leading to an unaltered dissolution profile during storage. A statistical design was conducted that revealed the critical process parameters affecting the taste masking efficiency and drug release. This study shows the successful application of solvent-free hot-melt coating in the development of a taste-masked and stable formulation. PMID:26621689

  9. Genetic Stability Developed for β-Carotene Synthesis in BR29 Rice Line Using Dihaploid Homozygosity

    PubMed Central

    Datta, Karabi; Sahoo, Gayetri; Krishnan, Sellappan; Ganguly, Moumita; Datta, Swapan K.

    2014-01-01

    Obtaining transgenic crop lines with stable levels of carotenoids is highly desirable. We addressed this issue by employing the anther culture technique to develop dihaploid lines containing genes involved in β-carotene metabolism. First, we used Agrobacterium- mediated transformation to develop primary transgenic plants containing the β-carotene biosynthetic genes, phytoene synthase (psy) and phytoene desaturase (crtI), which were engineered for expression and accumulation in the endosperm. Transgenic plants were recovered by selecting for the expression of the phosphomannose isomerase (pmi) gene. Dihaploid plants in addition to haploid and tetraploid plant were generated from anther cultures of these primary transgenic plants. In addition to anatomical features of stomata, pollen of different ploidy-plants, molecular analyses confirmed the stable integration of the genes in the anther culture-derived dihaploid plants, and the yellow color of the polished seeds indicated the accumulation of carotenoids in the endosperm. High performance liquid chromatography (HPLC) analysis of the carotenoid extract further confirmed the levels of β–carotene accumulation in the endosperms of the transgenic dihaploid rice seeds. PMID:24937154

  10. Transgenic rice plants expressing the antifungal AFP protein from Aspergillus giganteus show enhanced resistance to the rice blast fungus Magnaporthe grisea.

    PubMed

    Coca, María; Bortolotti, Cristina; Rufat, Mar; Peñas, Gisela; Eritja, Ramón; Tharreau, Didier; del Pozo, Alvaro Martinez; Messeguer, Joaquima; San Segundo, Blanca

    2004-01-01

    The Aspergillus giganteus antifungal protein (AFP), encoded by the afp gene, has been reported to possess in vitro antifungal activity against various economically important fungal pathogens, including the rice blast fungus Magnaporthe grisea. In this study, transgenic rice ( Oryza sativa ) constitutively expressing the afp gene was generated by Agrobacterium -mediated transformation. Two different DNA constructs containing either the afp cDNA sequence from Aspergillus or a chemically synthesized codon-optimized afp gene were introduced into rice plants. In both cases, the DNA region encoding the signal sequence from the tobacco AP24 gene was N-terminally fused to the coding sequence of the mature AFP protein. Transgenic rice plants showed stable integration and inheritance of the transgene. No effect on plant morphology was observed in the afp -expressing rice lines. The inhibitory activity of protein extracts prepared from leaves of afp plants on the in vitro growth of M. grisea indicated that the AFP protein produced by the trangenic rice plants was biologically active. Several of the T(2) homozygous afp lines were challenged with M. grisea in a detached leaf infection assay. Transformants exhibited resistance to rice blast at various levels. Altogether, the results presented here indicate that AFP can be functionally expressed in rice plants for protection against the rice blast fungus M. grisea. PMID:15159626

  11. Transgenic indica rice lines, expressing Brassica juncea Nonexpressor of pathogenesis-related genes 1 (BjNPR1), exhibit enhanced resistance to major pathogens.

    PubMed

    Sadumpati, Vijayakumar; Kalambur, Muralidharan; Vudem, Dashavantha Reddy; Kirti, Pulugurtha Bharadwaja; Khareedu, Venkateswara Rao

    2013-07-10

    Brassica juncea Nonexpressor of pathogenesis-related genes 1 (BjNPR1) has been introduced into commercial indica rice varieties by Agrobacterium-mediated genetic transformation. Transgenic rice plants were regenerated from the phosphinothricin-resistant calli obtained after co-cultivation with Agrobacterium strain LBA4404 harbouring Ti plasmid pSB111-bar-BjNPR1. Molecular analyses confirmed the stable integration and expression of BjNPR1 in various transgenic rice lines. Transgenes NPR1 and bar were stably inherited and disclosed co-segregation in subsequent generations in a Mendelian fashion. Homozygous transgenic rice lines expressing BjNPR1 protein displayed enhanced resistance to rice blast, sheath blight and bacterial leaf blight diseases. Rice transformants with higher levels of NPR1 revealed notable increases in plant height, panicle length, flag-leaf length, number of seeds/panicle and seed yield/plant as compared to the untransformed plants. The overall results amply demonstrate the profound impact of BjNPR1 in imparting resistance against major pathogens of rice. The multipotent BjNPR1, as such, seems promising as a prime candidate gene to fortify crop plants with durable resistance against various pathogens. PMID:23664883

  12. Problems associated with gene transfer and opportunities for microgravity environments

    NASA Astrophysics Data System (ADS)

    Tennessen, Daniel J.

    1997-01-01

    The method of crop improvement by gene transfer is becoming increasingly routine with transgenic foods and ornamental crops now being marketed to consumers. However, biological processes of plants, and the physical barriers of current protocols continue to limit the application of gene transfer in many commercial crops. The goal of this paper is to outline the current limitations of gene transfer and to hypothesize possible opportunities for use of microgravity to overcome such limitations. The limitations detailed in this paper include host-range specificity of Agrobacterium mediated transformation, probability of gene insertion, position effects of the inserted genes, gene copy number, stability of foreign gene expression in host plants, and regeneration of recalcitrant plant species. Microgravity offers an opportunity for gene transfer where cell growth kinetics, DNA synthesis, and genetic recombination rates can be altered. Such biological conditions may enhance the ability for recombination of reporter genes and other genes of interest to agriculture. Proposed studies would be useful for understanding instability of foreign gene expression and may lead to stable transformed plants. Other aspects of gene transfer in microgravity are discussed.

  13. Enhanced resistance to blister blight in transgenic tea (Camellia sinensis [L.] O. Kuntze) by overexpression of class I chitinase gene from potato (Solanum tuberosum).

    PubMed

    Singh, H Ranjit; Deka, Manab; Das, Sudripta

    2015-07-01

    Tea is the second most consumed beverage in the world. A crop loss of up to 43 % has been reported due to blister blight disease of tea caused by a fungus, Exobasidium vexans. Thus, it directly affects the tea industry qualitatively and quantitatively. Solanum tuberosum class I chitinase gene (AF153195) is a plant pathogenesis-related gene. It was introduced into tea genome via Agrobacterium-mediated transformation with hygromycin phosphotransferase (hpt) gene conferring hygromycin resistance as plant selectable marker. A total of 41 hygromycin resistant plantlets were obtained, and PCR analysis established 12 plantlets confirming about the stable integration of transgene in the plant genome. Real-time PCR detected transgene expression in four transgenic plantlets (T28, C57, C9, and T31). Resistance to biotrophic fungal pathogen, E. vexans, was tested by detached leaf infection assay of greenhouse acclimated plantlets. An inhibitory activity against the fungal pathogen was evident from the detached leaves from the transformants compared with the control. Fungal lesion formed on control plantlet whereas the transgenic plantlets showed resistance to inoculated fungal pathogen by the formation of hypersensitivity reaction area. This result suggests that constitutive expression of the potato class I chitinase gene can be exploited to improve resistance to fungal pathogen, E. vexans, in economical perennial plantation crop like tea. PMID:25772466

  14. Electron-Beam Induced Transformations of Layered Tin Dichalcogenides.

    PubMed

    Sutter, E; Huang, Y; Komsa, H-P; Ghorbani-Asl, M; Krasheninnikov, A V; Sutter, P

    2016-07-13

    By combining high-resolution transmission electron microscopy and associated analytical methods with first-principles calculations, we study the behavior of layered tin dichalcogenides under electron beam irradiation. We demonstrate that the controllable removal of chalcogen atoms due to electron irradiation, at both room and elevated temperatures, gives rise to transformations in the atomic structure of Sn-S and Sn-Se systems so that new phases with different properties can be induced. In particular, rhombohedral layered SnS2 and SnSe2 can be transformed via electron beam induced loss of chalcogen atoms into highly anisotropic orthorhombic layered SnS and SnSe. A striking dependence of the layer orientation of the resulting SnS-parallel to the layers of ultrathin SnS2 starting material, but slanted for transformations of thicker few-layer SnS2-is rationalized by a transformation pathway in which vacancies group into ordered S-vacancy lines, which convert via a Sn2S3 intermediate to SnS. Absence of a stable Sn2Se3 intermediate precludes this pathway for the selenides, hence SnSe2 always transforms into basal plane oriented SnSe. Our results provide microscopic insights into the transformation mechanism and show how irradiation can be used to tune the properties of layered tin chalcogenides for applications in electronics, catalysis, or energy storage. PMID:27336595

  15. Stable expression of a GFP-BSD fusion protein in transfected and blasticidin-selected B. bovis merozoites

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Babesia bovis is a tick-borne apicomplexan parasite that causes an acute disease in cattle. This study describes stable expression of an exogenous gfp-bsd gene in B. bovis transformed parasites. Cultured B. bovis infected erythrocytes of the biologically cloned Mo7 strain were transfected by electro...

  16. High-Order Energy Stable WENO Schemes

    NASA Technical Reports Server (NTRS)

    Yamaleev, Nail K.; Carpenter, Mark H.

    2008-01-01

    A new third-order Energy Stable Weighted Essentially NonOscillatory (ESWENO) finite difference scheme for scalar and vector linear hyperbolic equations with piecewise continuous initial conditions is developed. The new scheme is proven to be stable in the energy norm for both continuous and discontinuous solutions. In contrast to the existing high-resolution shock-capturing schemes, no assumption that the reconstruction should be total variation bounded (TVB) is explicitly required to prove stability of the new scheme. A rigorous truncation error analysis is presented showing that the accuracy of the 3rd-order ESWENO scheme is drastically improved if the tuning parameters of the weight functions satisfy certain criteria. Numerical results show that the new ESWENO scheme is stable and significantly outperforms the conventional third-order WENO finite difference scheme of Jiang and Shu in terms of accuracy, while providing essentially nonoscillatory solutions near strong discontinuities.

  17. Stable Isotope Enrichment Capabilities at ORNL

    SciTech Connect

    Egle, Brian; Aaron, W Scott; Hart, Kevin J

    2013-01-01

    The Oak Ridge National Laboratory (ORNL) and the US Department of Energy Nuclear Physics Program have built a high-resolution Electromagnetic Isotope Separator (EMIS) as a prototype for reestablishing a US based enrichment capability for stable isotopes. ORNL has over 60 years of experience providing enriched stable isotopes and related technical services to the international accelerator target community, as well as medical, research, industrial, national security, and other communities. ORNL is investigating the combined use of electromagnetic and gas centrifuge isotope separation technologies to provide research quantities (milligram to several kilograms) of enriched stable isotopes. In preparation for implementing a larger scale production facility, a 10 mA high-resolution EMIS prototype has been built and tested. Initial testing of the device has simultaneously collected greater than 98% enriched samples of all the molybdenum isotopes from natural abundance feedstock.

  18. Concentration of stable elements in food products

    SciTech Connect

    Montford, M.A.; Shank, K.E.; Hendricks, C.; Oakes, T.W.

    1980-01-01

    Food samples were taken from commercial markets and analyzed for stable element content. The concentrations of most stable elements (Ag, Al, As, Au, Ba, Br, Ca, Ce, Cl, Co, Cr, Cs, Cu, Fe, Hf, I, K, La, Mg, Mn, Mo, Na, Rb, Sb, Sc, Se, Sr, Ta, Th, Ti, V, Zn, Zr) were determined using multiple-element neutron activation analysis, while the concentrations of other elements (Cd, Hg, Ni, Pb) were determined using atomic absorption. The relevance of the concentrations found are noted in relation to other literature values. An earlier study was extended to include the determination of the concentration of stable elements in home-grown products in the vicinity of the Oak Ridge National Laboratory. Comparisons between the commercial and local food-stuff values are discussed.

  19. Stable isotope and elemental analysis in ants.

    PubMed

    Smith, Chris R; Tillberg, Chadwick V

    2009-07-01

    Over the past 20 yr, the use of stable isotopes to infer feeding ecology and the examination of how energetic and elemental exchanges are affected by and affect life (ecological stoichiometry) have gained momentum. The ecological diversity of ants makes them interesting models to explore dietary ecology and their role in food webs. Moreover, their ecological dominance in most habitats facilitates sampling. The protocol described here will produce samples adequate for submission to most labs that specialize in high-throughput analysis of stable isotopes; one should check with any particular lab for specific submission instructions. Note, however, that this protocol is designed specifically for the quantification of the natural abundance of stable isotopes; it does not cover the preparation of trace samples. PMID:20147207

  20. Low work function, stable thin films

    DOEpatents

    Dinh, Long N.; McLean, II, William; Balooch, Mehdi; Fehring, Jr., Edward J.; Schildbach, Marcus A.

    2000-01-01

    Generation of low work function, stable compound thin films by laser ablation. Compound thin films with low work function can be synthesized by simultaneously laser ablating silicon, for example, and thermal evaporating an alkali metal into an oxygen environment. For example, the compound thin film may be composed of Si/Cs/O. The work functions of the thin films can be varied by changing the silicon/alkali metal/oxygen ratio. Low work functions of the compound thin films deposited on silicon substrates were confirmed by ultraviolet photoelectron spectroscopy (UPS). The compound thin films are stable up to 500.degree. C. as measured by x-ray photoelectron spectroscopy (XPS). Tests have established that for certain chemical compositions and annealing temperatures of the compound thin films, negative electron affinity (NEA) was detected. The low work function, stable compound thin films can be utilized in solar cells, field emission flat panel displays, electron guns, and cold cathode electron guns.