Relevant bibliographies by topics / Genetic transformation

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Academic literature on the topic 'Genetic transformation'

Author: Grafiati

Published: 4 June 2021

Last updated: 1 August 2024

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Journal articles on the topic "Genetic transformation"

Donmez, Dicle, Ozhan Simsek, Tolga Izgu, Yildiz Aka Kacar, and Yesim Yalcin Mendi. "Genetic Transformation inCitrus." Scientific World Journal 2013 (2013): 1–8. http://dx.doi.org/10.1155/2013/491207.

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Citrus is one of the world’s important fruit crops. Recently, citrus molecular genetics and biotechnology work have been accelerated in the world. Genetic transformation, a biotechnological tool, allows the release of improved cultivars with desirable characteristics in a shorter period of time and therefore may be useful in citrus breeding programs.Citrustransformation has now been achieved in a number of laboratories by various methods.Agrobacterium tumefaciensis used mainly in citrus transformation studies. Particle bombardment, electroporation,A. rhizogenes, and a new method called RNA interference are used in citrus transformation studies in addition toA. tumefaciens. In this review, we illustrate how different gene transformation methods can be employed in different citrus species.

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De Bustos, A., R. Pérez, and N. Jouve. "Study of the homologous recombination genetic system to improve genetic transformation of wheat." Czech Journal of Genetics and Plant Breeding 41, Special Issue (July 31, 2012): 290–93. http://dx.doi.org/10.17221/6195-cjgpb.

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Gietz, R. Daniel, and Robin A. Woods. "Genetic Transformation of Yeast." BioTechniques 30, no. 4 (April 2001): 816–31. http://dx.doi.org/10.2144/01304rv02.

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Mathews, H., H. D. Wilde, R. E. Litz, and H. Y. Wetzstein. "GENETIC TRANSFORMATION OF MANGO." Acta Horticulturae, no. 341 (May 1993): 93–97. http://dx.doi.org/10.17660/actahortic.1993.341.8.

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Moss, Robert. "Genetic Transformation of Bacteria." American Biology Teacher 53, no. 3 (March 1, 1991): 179–80. http://dx.doi.org/10.2307/4449256.

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Bhatia, C. R., Patricia Viegas, Anjali Bhagwat, Helena Mathews, and N. K. Notani. "Genetic transformation of plants." Proceedings / Indian Academy of Sciences 96, no. 2 (June 1986): 79–112. http://dx.doi.org/10.1007/bf03053326.

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Ribas, Alessandra Ferreira, Luiz Filipe Protasio Pereira, and Luiz Gonzaga E. Vieira. "Genetic transformation of coffee." Brazilian Journal of Plant Physiology 18, no. 1 (March 2006): 83–94. http://dx.doi.org/10.1590/s1677-04202006000100007.

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In the last 15 years, considerable advances were made in coffee genetic transformation. Different research groups in the world have been able to transform coffee with genes for insect resistance, decaffeinated coffee, herbicide resistance and control of fruit maturation. Although the majority of the research is still limited to laboratory and greenhouse studies, initial field tests with transformed coffee are beginning to appear in the literature. In this review we provide an update on the state of coffee genetic transformation, presenting technical aspects related to tissue culture systems, strategies for selection and transformation with particle bombardment, as well as the use of Agrobacterium tumefaciens. We also discuss the potential applications of this technology, taking into consideration the benefits, the possible environmental risks, as well as market and consumer issues.

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Langeveld, S. A., S. Marinova, M. M. Gerrits, A. F. L. M. Derks, and P. M. Boonekamp. "GENETIC TRANSFORMATION OF LILY." Acta Horticulturae, no. 430 (December 1997): 290. http://dx.doi.org/10.17660/actahortic.1997.430.43.

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He, Liya, Jiao Feng, Sha Lu, Zhiwen Chen, Chunmei Chen, Ya He, Xiuwen Yi, and Liyan Xi. "Genetic transformation of fungi." International Journal of Developmental Biology 61, no. 6-7 (2017): 375–81. http://dx.doi.org/10.1387/ijdb.160026lh.

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Tsuda, Masataka, Mikio Karita, and Teruo Nakazawa. "Genetic Transformation inHelicobacter pylori." Microbiology and Immunology 37, no. 1 (January 1993): 85–89. http://dx.doi.org/10.1111/j.1348-0421.1993.tb03184.x.

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Dissertations / Theses on the topic "Genetic transformation"

Zainuddin. "Genetic transformation of wheat (Triticum aestivum L.)." Title page, Contents and Abstract only, 2000. http://web4.library.adelaide.edu.au/theses/09APSP/09apspz21.pdf.

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Bibliography: leaves 127-151. The successful application of genetic engineering in wheat is dependent on the availability of suitable tissue culture and transformation methods. The primary object of this project was the development of these technologies using elite Australian wheat varieties.

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Button, Eric A. "Regulation of T-DNA gene 7." Thesis, University of British Columbia, 1987. http://hdl.handle.net/2429/26177.

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The purpose of this study was two-fold. The first objective was to determine if Saccharomyces cerevisiae is a useful system for investigating the expression of T-DNA (it takes several months to obtain sufficient bacteria-free transformed plant tissue to investigate T-DNA transcription). A short fragment of T-DNA carrying T-DNA gene 7 was cloned into a yeast plasmid in an attempt to investigate the expression of gene 7 in yeast. The second objective was to determine the significance of a heat shock related sequence identified in the 5¹ region of T-DNA gene 7. Primer extension analysis, SI nuclease mapping, and Northern hybridizations indicate that transcription of T-DNA gene 7 in yeast is different from that of transcription of gene 7 in crown gall tumors. Transcription is different because the distance between the TATA box and the transcription initiation sites must be at least 40 nucleotides in yeast. Therefore, Saccharomyces cerevisiae does not appear to be a useful system for investigating the expression of T-DNA. Crown gall tumors were subjected to a number of stress agents, including heat shock, to determine the significance of the heat shock related sequence identified in gene 7. Primer extension analyses indicate that only cadmium and mercury have a significant effect on the expression of T-DNA gene 7. Although gene 7 responds to cadmium and mercury, the increase in transcription does not appear to be heat shock or metallothionein related, indicating that another mechanism is involved in the enhanced transcription of T-DNA gene 7 in crown gall tumors.
Medicine, Faculty of
Medical Genetics, Department of
Graduate

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Tor, Mahmut. "Genetic transformation of yam (Dioscorea)." Thesis, Imperial College London, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.267504.

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Gartland, Kevan M. A. "Studies on plant genetic transformation." Thesis, University of Nottingham, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.236507.

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Fryer, Shirley Anne. "Genetic transformation of oilseed rape." Thesis, University of Wolverhampton, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.317928.

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Chen, Dong Fang. "Genetic transformation in the Gramineae." Thesis, Open University, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.293321.

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Soloki, Mahmod. "Genetic transformation of grape somatic embryos." Thesis, University of Nottingham, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.387659.

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Faria, Maria José Sparça Salles de. "Red raspberry transformation using agrobacterium." Thesis, McGill University, 1993. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=69522.

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Regeneration and transformation protocols for 'Comet' red raspberry were optimized with the purpose of making the Agrobacterium-mediated gene transfer system efficient for this crop. Adventitious shoot regeneration from leaf discs was improved using explants 10 mm in diameter and transferring to fresh medium at the fourth week of incubation. Additions of liquid medium to solid medium during incubation decreased regeneration and attempts to release the suppressive influence of larger shoots on initials (apical dominance) did not succeed. The presence of claforan did not affect shoot regeneration, but inoculations with Agrobacterium and the presence of kanamycin decreased regeneration moderately or considerably, respectively. The threshold for kanamycin concentration for screening for kanamycin resistant transformed raspberry tissue was 30 to 40 mg l$ sp{-1}.$ The best co-incubation interval between wild-type Agrobacterium and 'Comet' leaf discs ranged from 2 days for highly virulent strains to 3 or more days for moderate to low virulent strains. Among several wild-type strains, C58 was chosen as the most appropriate partially because a disarmed form was commercially available for use as a non-oncogenic vector for transformation of red raspberry.
The binary plasmid pBI121 containing the marker genes NPTII and GUS encoding kanamycin resistance and $ beta$-glucuronidase activity, respectively, was successfully introduced into the Agrobacterium strain LBA4404, which is a disarmed C58 derivative. Transformation of 'Comet' red raspberry was apparently achieved by inoculating leaf disc explants with LBA4404 containing pBI121. The probable integration and expression of the foreign genes into the plant cells were confirmed by screening for kanamycin resistance, GUS assays and Southern blot analyses. This transformation system appears to be effective and may be useful in further studies on red raspberry for both introduction of genes for desirable agronomic traits and basic studies of gene expression.

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Robson, Julia. "The construction of an expression vector for the transformation of the grape chloroplast genome." Thesis, Stellenbosch : Stellenbosch University, 2003. http://hdl.handle.net/10019.1/53621.

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Thesis (MSc)--University of Stellenbosch, 2003.
ENGLISH ABSTRACT: The genetic information of plants is found in the nucleus, the mitochondria, and the plastids. The DNA of plastids is comprised of multiple copies of a double-stranded, circular, prokaryoticallyderived genome of -150 kb. The genome equivalents of plastid organelles in higher plant cells are an attractive target for genetic engineering as high protein expression levels are readily obtained due to the high genome copy number per organelle. The resultant proteins are contained within the plastid organelle and the corresponding transgenes are inherited, in most crop plants, uniparentally, preventing pollen transmission of DNA. Plastid transformation involves the uniform modification of all the plastid genome copies, a process facilitated by homologous recombination and the non-Mendelian segregation of plastids upon cell division. The plastid genomes are in a continuous state of inter- and intra-molecular exchange due to their common genetic complement. This enables the site-specific integration of any piece of DNA flanked by plastid targeting sequences, via homologous recombination. The attainment of homoplasmy, where all genomes are transformed, requires the inclusion of a plastid-specific selectable marker. Selective pressure favouring the propagation of the transformed genome copies, as well as the random segregation of plastids upon cell division, make it feasible to acquire uniformity and hence genetic stability. From this, a complete transplastomie line is obtained where all plastid genome copies present are transgenic, having eliminated all wild-type genome copies. The prokaryotic nature of the chloroplast genetic system enables expression of multiple proteins from polycistronic mRNAs, allowing the introduction of entire operons in a single transformation. Expression cassettes in vectors thus include single regulatory elements of plastid origin, and harbour genes encoding selectable and screenable markers, as well as one or more genes of interest. Each coding region is preceded by an appropriate translation control region to ensure efficient translation from the polycistronic mRNA. The function of a plastid transformation vector is to enable transfer and stable integration of foreign genes into the chloroplast genomes of higher plants. The expression vector constructed in this research is specific for the transformation of the grape chloroplast genome. Vitis vinifera L., from the family, Vitaceae, is the choice species for the production of wine and therefore our target for plastid transformation. All chloroplast derived regulatory elements and sequences included in the vector thus originated from this species.
AFRIKAANSE OPSOMMING: Die genetiese inligting van plante word gevind in die kern, die mitochondria, en die plastiede. Die DNA van plastiede bestaan uit veelvuldige kopieë van 'n ~ 150 kb dubbelstring, sirkulêre genoom van prokariotiese oorsprong. Die genoomekwivalente van plastiede in hoër plante is 'n aantreklike teiken vir genetiese manipulering, aangesien die hoë genoom kopiegetal per organel dit moontlik maak om gereeld hoë vlakke van proteïenuitdrukking te verkry. Hierdie proteïene word tot die plastied beperk, en die ooreenstemmende transgene word in die meeste plante sitoplasmies oorgeërf, sonder die oordrag van DNA deur die stuifmeel. Plastied transformasie behels die uniforme modifikasie van al die plastied genoomkopieë, 'n proses wat deur homoloë rekombinasie en die nie-Mendeliese segregasie van plastiede tydens seldeling gefasiliteer word. As gevolg van die gemeenskaplike genetiese komplement, vind aanhoudende interen intra-molekulêre uitruiling van plastiedgenome plaas. Dit maak die setel-spesifieke integrasie, via homoloë rekombinasie, van enige stuk DNA wat deur plastied teikenvolgordes begrens word, moontlik. Vir die verkrying van homoplasmie, waar alle genome getransformeer is, word die insluiting van 'n plastiedspesifieke selekteerbare merker benodig. Seleksiedruk wat die vermeerdering van die getransformeerde genoomkopieë bevoordeel, en die lukrake segregasie van plastiede tydens seldeling, maak dit moontlik om genetiese stabiliteit en uniformiteit van die genoom te verkry. Dit kan op sy beurt tot die verkryging van 'n volledige transplastomiese lyn lei, waar alle aanwesige plastiedgenome transgenies is, en wilde tipe genoomkopieë geëlimineer is. Die prokariotiese aard van die chloroplas genetiese sisteem maak die uitdrukking van veelvuldige proteïene vanaf polisistroniese mRNAs moontlik, wat die toevoeging van volledige operons in 'n enkele transformasie toelaat. Uitdrukkingskassette in vektore bevat dus enkel regulatoriese elemente van plastied oorsprong, gene wat kodeer vir selekteerbare en sifbare merkers, asook een of meer gene van belang (teikengene). Voor elke koderingsstreek, is daar ook 'n toepaslike translasie beheerstreek om doeltreffende translasie vanaf die polisistroniese mRNA te verseker. Die funksie van 'n plastied transformasie vektor is om die oordrag en stabiele integrasie van transgene in chloroplasgenome van hoër plante moontlik te maak. Die uitdrukkingsvektor wat in hierdie studie gekonstrueer is, is spesifiek vir die transformasie van die druif chloroplasgenoom. Vitis vinifera L., van die familie Vitaceae, is die voorkeur species vir die produksie van wyn, en daarom die teiken vir plastied transformasie. Alle chloroplast-afgeleide regulatoriese elemente en volgordes wat in hierdie vektor ingesluit is, het huloorsprong vanaf VUis vinifera L.

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Cook, Marisa Anne. "Replicons derived from endogenously isolated plasmids used to classify plasmids occurring in marine sediment bacteria." Thesis, Georgia Institute of Technology, 2001. http://hdl.handle.net/1853/25736.

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Books on the topic "Genetic transformation"

Linskens, H. F., and J. F. Jackson. Genetic transformation of plants. Berlin: Springer, 2010.

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Walden, R. Genetic transformation in plants. Englewood Cliffs, N.J: Prentice Hall, 1989.

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Stewart, C. Neal. Plant transformation technologies. Ames, Iowa: Wiley-Blackwell, 2011.

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Jackson, J. F., and H. F. Linskens, eds. Genetic Transformation of Plants. Berlin, Heidelberg: Springer Berlin Heidelberg, 2003. http://dx.doi.org/10.1007/978-3-662-07424-4.

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O, Butler L., Harwood Colin R, and Moseley B. E. B, eds. Genetic transformation and expression. Andover, Hants [England]: Intercept, 1990.

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O, Butler L., Harwood Colin, and Moseley B. E. B, eds. Genetic transformation and expression. Andover: Intercept, 1989.

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Wellington, E. M. H. 1954- and Elsas, J. D. van 1951-, eds. Genetic interactions between microorgamisms in the natural environment: Gene transfer in nature. Manchester: Manchester University Press, 1992.

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Koch-Brandt, Claudia. Gentransfer: Prinzipien, Experimente, Anwendung bei Saügern. Stuttgart: G. Thieme, 1993.

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1957-, Draper John, ed. Plantgenetic transformation and gene expression: A laboratory manual. Oxford: Blackwell Scientific, 1988.

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Fryer, Shirley Anne. Genetic transformation of oilseed rape. Wolverhampton: University of Wolverhampton, 1992.

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Book chapters on the topic "Genetic transformation"

Birge, Edward A. "Genetic Transformation." In Bacterial and Bacteriophage Genetics, 199–219. New York, NY: Springer New York, 1988. http://dx.doi.org/10.1007/978-1-4757-1995-6_8.

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Birge, Edward A. "Genetic Transformation." In Bacterial and Bacteriophage Genetics, 257–76. New York, NY: Springer New York, 1994. http://dx.doi.org/10.1007/978-1-4757-2328-1_10.

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Kroth, Peter G. "Genetic Transformation." In Protein Targeting Protocols, 257–67. Totowa, NJ: Humana Press, 2007. http://dx.doi.org/10.1007/978-1-59745-466-7_17.

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Mondal, Tapan Kumar. "Genetic Transformation." In Breeding and Biotechnology of Tea and its Wild Species, 85–92. New Delhi: Springer India, 2014. http://dx.doi.org/10.1007/978-81-322-1704-6_5.

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Birge, Edward A. "Genetic Transformation." In Bacterial and Bacteriophage Genetics, 315–39. New York, NY: Springer New York, 2000. http://dx.doi.org/10.1007/978-1-4757-3258-0_10.

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Peck, Stewart B., Carol C. Mapes, Netta Dorchin, John B. Heppner, Eileen A. Buss, Gustavo Moya-Raygoza, Marjorie A. Hoy, et al. "Genetic Transformation." In Encyclopedia of Entomology, 1597–99. Dordrecht: Springer Netherlands, 2008. http://dx.doi.org/10.1007/978-1-4020-6359-6_1062.

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Mondal, Tapan Kumar. "Genetic Transformation." In Tea: Genome and Genetics, 127–38. Singapore: Springer Singapore, 2020. http://dx.doi.org/10.1007/978-981-15-8868-6_5.

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Jacobsen, Hans-Jörg. "Genetic Transformation." In Developments in Plant Breeding, 125–32. Dordrecht: Springer Netherlands, 1999. http://dx.doi.org/10.1007/978-94-015-9211-6_5.

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Bilang, Roland, Johannes Fütterer, and Christof Sautter. "Transformation of Cereals." In Genetic Engineering, 113–57. Boston, MA: Springer US, 1999. http://dx.doi.org/10.1007/978-1-4615-4707-5_7.

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Xu, Jun-Wei. "Genetic Transformation System." In Compendium of Plant Genomes, 165–76. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-75710-6_9.

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Conference papers on the topic "Genetic transformation"

de França, Fabrício Olivetti. "Transformation-interaction-rational representation for symbolic regression." In GECCO '22: Genetic and Evolutionary Computation Conference. New York, NY, USA: ACM, 2022. http://dx.doi.org/10.1145/3512290.3528695.

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Aldeia, Guilherme Seidyo Imai, and Fabrício Olivetti de França. "Interaction-transformation evolutionary algorithm with coefficients optimization." In GECCO '22: Genetic and Evolutionary Computation Conference. New York, NY, USA: ACM, 2022. http://dx.doi.org/10.1145/3520304.3533987.

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Mayer, Benjamin E., and Kay Hamacher. "Stochastic tunneling transformation during selection in genetic algorithm." In GECCO '14: Genetic and Evolutionary Computation Conference. New York, NY, USA: ACM, 2014. http://dx.doi.org/10.1145/2576768.2598243.

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Ahn, Eun Yeong, Tracy Mullen, and John Yen. "Finding feature transformation functions using genetic algorithm." In the 12th annual conference comp. New York, New York, USA: ACM Press, 2010. http://dx.doi.org/10.1145/1830761.1830862.

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Pan, Shuaiqun, Diederick Vermetten, Manuel López-Ibáñez, Thomas Bäck, and Hao Wang. "Transfer Learning of Surrogate Models via Domain Affine Transformation." In GECCO '24: Genetic and Evolutionary Computation Conference. New York, NY, USA: ACM, 2024. http://dx.doi.org/10.1145/3638529.3654032.

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Narinç, Doğan, and Ali Aygün. "A non parametric data transformation technique for quantitative genetic analyses: The rank transformation." In II. INTERNATIONAL CONFERENCE ON ADVANCES IN NATURAL AND APPLIED SCIENCES: ICANAS 2017. Author(s), 2017. http://dx.doi.org/10.1063/1.4981708.

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Spagnolo, Nicolo, Enrico Maiorino, Chiara Vitelli, Marco Bentivegna, Andrea Crespi, Roberta Ramponi, Paolo Mataloni, Roberto Osellame, and Fabio Sciarrino. "Genetic algorithms to learn an unknown linear transformation." In 2017 Conference on Lasers and Electro-Optics Europe & European Quantum Electronics Conference (CLEO/Europe-EQEC). IEEE, 2017. http://dx.doi.org/10.1109/cleoe-eqec.2017.8087443.

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Faridmoayer, Sogol, Mohammadreza Sharbaf, and Shekoufeh Kolahdouz-Rahimi. "Optimization of model transformation output using genetic algorithm." In 2017 IEEE 4th International Conference on Knowledge-Based Engineering and Innovation (KBEI). IEEE, 2017. http://dx.doi.org/10.1109/kbei.2017.8324973.

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Li, Minghui, Kwok Shun Ho, and Gordon Hayward. "Beamspace transformation for data reduction using genetic algorithms." In 2009 IEEE International Ultrasonics Symposium. IEEE, 2009. http://dx.doi.org/10.1109/ultsym.2009.5442004.

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Ait ElHara, Ouassim, Anne Auger, and Nikolaus Hansen. "Permuted Orthogonal Block-Diagonal Transformation Matrices for Large Scale Optimization Benchmarking." In GECCO '16: Genetic and Evolutionary Computation Conference. New York, NY, USA: ACM, 2016. http://dx.doi.org/10.1145/2908812.2908937.

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Reports on the topic "Genetic transformation"

Voth, Wayne. Genetic Transformation Among Azotobacter Species. Portland State University Library, January 2000. http://dx.doi.org/10.15760/etd.2613.

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Seger, Yvette R. Genetic Requirements for the Transformation of Human Cells. Fort Belvoir, VA: Defense Technical Information Center, July 2002. http://dx.doi.org/10.21236/ada410207.

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Seger, Yvette. Genetic Requirements for the Transformation of Human Cells. Fort Belvoir, VA: Defense Technical Information Center, July 2004. http://dx.doi.org/10.21236/ada429117.

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Seger, Yvette M. Genetic Requirements for the Transformation of Human Cells. Fort Belvoir, VA: Defense Technical Information Center, July 2003. http://dx.doi.org/10.21236/ada418793.

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Gera, Abed, Abed Watad, P. Ueng, Hei-Ti Hsu, Kathryn Kamo, Peter Ueng, and A. Lipsky. Genetic Transformation of Flowering Bulb Crops for Virus Resistance. United States Department of Agriculture, January 2001. http://dx.doi.org/10.32747/2001.7575293.bard.

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Objectives. The major aim of the proposed research was to establish an efficient and reproducible genetic transformation system for Easter lily and gladiolus using either biolistics or Agrobacterium. Transgenic plants containing pathogen-derived genes for virus resistance were to be developed and then tested for virus resistance. The proposal was originally aimed at studying cucumber mosaic virus (CMV) resistance in plants, but studies later included bean yellow mosaic virus (BYMV). Monoclonal antibodies were to be tested to determine their effectiveness in interning with virus infection and vector (aphid) transmission. Those antibodies that effectively interfered with virus infection and transmission were to be cloned as single chain fragments and used for developing transgenic plants with the potential to resist virus infection. Background to the topic. Many flower crops, as lily and gladiolus are propagated vegetatively through bulbs and corms, resulting in virus transmission to the next planting generation. Molecular genetics offers the opportunity of conferring transgene-mediated disease resistance to flower crops that cannot be achieved through classical breeding. CMV infects numerous plant species worldwide including both lilies and gladioli. Major conclusions, solutions and achievements. Results from these for future development of collaborative studies have demonstrated the potential transgenic floral bulb crops for virus resistance. In Israel, an efficient and reproducible genetic transformation system for Easter lily using biolistics was developed. Transient as well as solid expression of GUS reporter gene was demonstrated. Putative transgenic lily plantlets containing the disabled CMV replicase transgene have been developed. The in vitro ability of monoclonal antibodies (mAbs) against CMV to neutralize virus infectivity and block virus transmission by M. persicae were demonstrated. In the US, transgenic Gladiolus plants containing either the BYMV coat protein or antisense coat protein genes have been developed and some lines were found to be virus resistant. Long-term expression of the GUS reporter gene demonstrated that transgene silencing did not occur after three seasons of dormancy in the 28 transgenic Gladiolus plants tested. Selected monoclonal antibody lines have been isolated, cloned as single chain fragments and are being used in developing transgenic plants with CMV resistance. Ornamental crops are multi-million dollar industries in both Israel and the US. The increasing economic value of these floral crops and the increasing ban numerous pesticides makes it more important than ever that alternatives to chemical control of pathogens be studied to determine their possible role in the future. The cooperation resulted in the objectives being promoted at national and international meetings. The cooperation also enabled the technology transfer between the two labs, as well as access to instrumentation and specialization particular to the two labs.

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Gray, Dennis, and Victor Gaba. Genotype, Explant and Growth Regulator Effects in the Determination of Adventitious Regeneratin in Curcurbits, in Aid of Genetic Transformation. United States Department of Agriculture, June 1992. http://dx.doi.org/10.32747/1992.7561060.bard.

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The objective of this study was to gain an understanding of the in vitro regeneration process in watermelon and melon to enable the development of genetic transformation systems. The objectives were met and additional progress, unplanned during the original proposal, was made. Organogenic regeneration in vitro was studied in both melon and watermelon. Genotype played a significant role in regeneration. In melon, epidermal cells were responsible for most regeneration. Methods to obtain in vitro-derived watermelon tetraploids, needed for seedless varieties, were developed. The culture systems were refined so that they could be routinely used for transformation. Particle guns were constructed and Agrobacterium strains were obtained to study the effect of transformation procedures on culture system performance, allowing refinement of transformation protocols. The culture systems were shown to enable the stable transformation of both crops, allowing their future use for insertion of agriculturally-important genes. In addition, we showed that shoot apical meristems might be suitable target tissue for transformation and allow a wider range of genotypes to be used, which is needed for crops as diverse as cucurbits.

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Norelli, John L., Moshe Flaishman, Herb Aldwinckle, and David Gidoni. Regulated expression of site-specific DNA recombination for precision genetic engineering of apple. United States Department of Agriculture, March 2005. http://dx.doi.org/10.32747/2005.7587214.bard.

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Objectives: The original objectives of this project were to: 1) evaluate inducible promoters for the expression of recombinase in apple (USDA-ARS); 2) develop alternative selectable markers for use in apple to facilitate the positive selection of gene excision by recombinase (Cornell University); 3) compare the activity of three different recombinase systems (Cre/lox, FLP/FRT, and R/RS)in apple using a rapid transient assay (ARO); and 4) evaluate the use of recombinase systems in apple using the best promoters, selectable markers and recombinase systems identified in 1, 2 and 3 above (Collaboratively). Objective 2 was revised from the development alternative selectable markers, to the development of a marker-free selection system for apple. This change in approach was taken due to the inefficiency of the alternative markers initially evaluated in apple, phosphomannose-isomerase and 2-deoxyglucose-6-phosphate phosphatase, and the regulatory advantages of a marker-free system. Objective 3 was revised to focus primarily on the FLP/FRT recombinase system, due to the initial success obtained with this recombinase system. Based upon cooperation between researchers (see Achievements below), research to evaluate the use of the FLP recombinase system under light-inducible expression in apple was then conducted at the ARO (Objective 4). Background: Genomic research and genetic engineering have tremendous potential to enhance crop performance, improve food quality and increase farm profits. However, implementing the knowledge of genomics through genetically engineered fruit crops has many hurdles to be overcome before it can become a reality in the orchard. Among the most important hurdles are consumer concerns regarding the safety of transgenics and the impact this may have on marketing. The goal of this project was to develop plant transformation technologies to mitigate these concerns. Major achievements: Our results indicate activity of the FLP\FRTsite-specific recombination system for the first time in apple, and additionally, we show light- inducible activation of the recombinase in trees. Initial selection of apple transformation events is conducted under dark conditions, and tissue cultures are then moved to light conditions to promote marker excision and plant development. As trees are perennial and - cross-fertilization is not practical, the light-induced FLP-mediated recombination approach shown here provides an alternative to previously reported chemically induced recombinase approaches. In addition, a method was developed to transform apple without the use of herbicide or antibiotic resistance marker genes (marker free). Both light and chemically inducible promoters were developed to allow controlled gene expression in fruit crops. Implications: The research supported by this grant has demonstrated the feasibility of "marker excision" and "marker free" transformation technologies in apple. The use of these safer technologies for the genetic enhancement of apple varieties and rootstocks for various traits will serve to mitigate many of the consumer and environmental concerns facing the commercialization of these improved varieties.

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Ron, Eliora, and Eugene Eugene Nester. Global functional genomics of plant cell transformation by agrobacterium. United States Department of Agriculture, March 2009. http://dx.doi.org/10.32747/2009.7695860.bard.

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The aim of this study was to carry out a global functional genomics analysis of plant cell transformation by Agrobacterium in order to define and characterize the physiology of Agrobacterium in the acidic environment of a wounded plant. We planed to study the proteome and transcriptome of Agrobacterium in response to a change in pH, from 7.2 to 5.5 and identify genes and circuits directly involved in this change. Bacteria-plant interactions involve a large number of global regulatory systems, which are essential for protection against new stressful conditions. The interaction of bacteria with their hosts has been previously studied by genetic-physiological methods. We wanted to make use of the new capabilities to study these interactions on a global scale, using transcription analysis (transcriptomics, microarrays) and proteomics (2D gel electrophoresis and mass spectrometry). The results provided extensive data on the functional genomics under conditions that partially mimic plant infection and – in addition - revealed some surprising and significant data. Thus, we identified the genes whose expression is modulated when Agrobacterium is grown under the acidic conditions found in the rhizosphere (pH 5.5), an essential environmental factor in Agrobacterium – plant interactions essential for induction of the virulence program by plant signal molecules. Among the 45 genes whose expression was significantly elevated, of special interest is the two-component chromosomally encoded system, ChvG/I which is involved in regulating acid inducible genes. A second exciting system under acid and ChvG/Icontrol is a secretion system for proteins, T6SS, encoded by 14 genes which appears to be important for Rhizobium leguminosarum nodule formation and nitrogen fixation and for virulence of Agrobacterium. The proteome analysis revealed that gamma aminobutyric acid (GABA), a metabolite secreted by wounded plants, induces the synthesis of an Agrobacterium lactonase which degrades the quorum sensing signal, N-acyl homoserine lactone (AHL), resulting in attenuation of virulence. In addition, through a transcriptomic analysis of Agrobacterium growing at the pH of the rhizosphere (pH=5.5), we demonstrated that salicylic acid (SA) a well-studied plant signal molecule important in plant defense, attenuates Agrobacterium virulence in two distinct ways - by down regulating the synthesis of the virulence (vir) genes required for the processing and transfer of the T-DNA and by inducing the same lactonase, which in turn degrades the AHL. Thus, GABA and SA with different molecular structures, induce the expression of these same genes. The identification of genes whose expression is modulated by conditions that mimic plant infection, as well as the identification of regulatory molecules that help control the early stages of infection, advance our understanding of this complex bacterial-plant interaction and has immediate potential applications to modify it. We expect that the data generated by our research will be used to develop novel strategies for the control of crown gall disease. Moreover, these results will also provide the basis for future biotechnological approaches that will use genetic manipulations to improve bacterial-plant interactions, leading to more efficient DNA transfer to recalcitrant plants and robust symbiosis. These advances will, in turn, contribute to plant protection by introducing genes for resistance against other bacteria, pests and environmental stress.

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Tzfira, Tzvi, Michael Elbaum, and Sharon Wolf. DNA transfer by Agrobacterium: a cooperative interaction of ssDNA, virulence proteins, and plant host factors. United States Department of Agriculture, December 2005. http://dx.doi.org/10.32747/2005.7695881.bard.

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Agrobacteriumtumefaciensmediates genetic transformation of plants. The possibility of exchanging the natural genes for other DNA has led to Agrobacterium’s emergence as the primary vector for genetic modification of plants. The similarity among eukaryotic mechanisms of nuclear import also suggests use of its active elements as media for non-viral genetic therapy in animals. These considerations motivate the present study of the process that carries DNA of bacterial origin into the host nucleus. The infective pathway of Agrobacterium involves excision of a single-stranded DNA molecule (T-strand) from the bacterial tumor-inducing plasmid. This transferred DNA (T-DNA) travels to the host cell cytoplasm along with two virulence proteins, VirD2 and VirE2, through a specific bacteriumplant channel(s). Little is known about the precise structure and composition of the resulting complex within the host cell and even less is known about the mechanism of its nuclear import and integration into the host cell genome. In the present proposal we combined the expertise of the US and Israeli labs and revealed many of the biophysical and biological properties of the genetic transformation process, thus enhancing our understanding of the processes leading to nuclear import and integration of the Agrobacterium T-DNA. Specifically, we sought to: I. Elucidate the interaction of the T-strand with its chaperones. II. Analyzing the three-dimensional structure of the T-complex and its chaperones in vitro. III. Analyze kinetics of T-complex formation and T-complex nuclear import. During the past three years we accomplished our goals and made the following major discoveries: (1) Resolved the VirE2-ssDNA three-dimensional structure. (2) Characterized VirE2-ssDNA assembly and aggregation, along with regulation by VirE1. (3) Studied VirE2-ssDNA nuclear import by electron tomography. (4) Showed that T-DNA integrates via double-stranded (ds) intermediates. (5) Identified that Arabidopsis Ku80 interacts with dsT-DNA intermediates and is essential for T-DNA integration. (6) Found a role of targeted proteolysis in T-DNA uncoating. Our research provide significant physical, molecular, and structural insights into the Tcomplex structure and composition, the effect of host receptors on its nuclear import, the mechanism of T-DNA nuclear import, proteolysis and integration in host cells. Understanding the mechanical and molecular basis for T-DNA nuclear import and integration is an essential key for the development of new strategies for genetic transformation of recalcitrant plant species. Thus, the knowledge gained in this study can potentially be applied to enhance the transformation process by interfering with key steps of the transformation process (i.e. nuclear import, proteolysis and integration). Finally, in addition to the study of Agrobacterium-host interaction, our research also revealed some fundamental insights into basic cellular mechanisms of nuclear import, targeted proteolysis, protein-DNA interactions and DNA repair.

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Stern, David, and Gadi Schuster. Manipulating Chloroplast Gene Expression: A Genetic and Mechanistic Analysis of Processes that Control RNA Stability. United States Department of Agriculture, June 2004. http://dx.doi.org/10.32747/2004.7586541.bard.

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New potential for engineering chloroplasts to express novel traits has stimulated research into relevant techniques and genetic processes, including plastid transformation and gene regulation. This BARD-funded research dealt with the mechanisms that influence chloroplast RNA accumulation, and thus gene expression. Previous work on cpRNA catabolism has elucidated a pathway initiated by endonucleolytic cleavage, followed by polyadenylation and exonucleolytic degradation. A major player in this process is the nucleus-encoded exoribo-nuclease/polymerase polynucleotide phosphorylase (PNPase). Biochemical characterization of PNPase has revealed a modular structure that controls its RNA synthesis and degradation activities, which in turn are responsive to the phosphate (P) concentration. During the funding period, new insights emerged into the molecular mechanism of RNA metabolism in the chloroplast and cyanobacteria, suggesting strategies for improving agriculturally-important plants or plants with novel introduced traits.

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