Progress 10/05/10 to 10/04/15
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Project Director no longer employed with the University.
Publications
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Progress 01/01/13 to 09/30/13
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The Project Director is no longer employed with the university
Publications
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: 1. Opuntia improvement. Results of our currently funded research on commercial lines of Opuntia ficus indica are being disseminated through the D'Arrigo Bros. company, which funds this research. This occurs once or twice each year at international meetings. 2. Cucumis improvement.The results of cantalope improvement research was published this year 3. Cotton improvement. Identification of a possible embryo lethal mutation in G. hirsutum, Texas Marker-1, a genetic standard for Gossypium. Arnold, Marianne, Gould JH. 2012. Abnormality in Cotton (Gossypium hirsutum Texas Marker-1) germination that is similar to an Arabidopsis mutant. Plant Biology 2012, Austin TX July 20-25, 2012. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
1. Opuntia improvement. At this point, we have results indicating we have stably transformed tissue. We are also getting closer to obtaining somatic embryos. Results will be presented to the D'Arrigo Bros. Board meeting this March. 2. Cucumis Improvement. The melon improvement community has been very positive about our addition to information on melon transformation and the trials with genes to improve beta-carotene content in breeder lines. 3. Cotton. Identification & characterization of a possible embryo lethal mutation in one of the Gossypium hirsutum is continuing. Lethality appears to be mediated by temperature of the environment in which the seed develops.
Publications
- Ren Y, Bang H, Lee EJ, Gould JH, Rathore K, Patil BS, Crosby KM. 2013. Levels of phytoene and beta- carotene in transgenic honeydew melon (Cucumis melo L. inodorus) Plant Cell Tissue and Organ Culture PCTO-D-12-05627R2.
- Ren Y, Bang H, Gould JH, Rathore K, Patil S, Crosby KM. 2012. Shoot regeneration and ploidy variation in tissue culture of honeydew melon (Cucumis melo L. inodorus). In Vitro Cellular and Developmental Biology Plant. Accepted 11-8-2012 IVPL-D-12-00145R3
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: Melon: Agrobacterium-based transformation procedures were successfully developed for western cantaloupe and honeydew melons for use in increasing the carotene content of the melon's flesh. Transgenic lines of the elite Honeydew '150' were created using watermelon phytoene synthase (PSY) and/or lycopene beta-cyclase (Lcy-B) genes driven by a tomato polygalacturonase (PG) promotor. PARTICIPANTS: Two research collaborations were dropped due to lack of funding. Cactus: Fang Wang, Meditith Turnipseed, Peter Felker (D'Arrigo Bros. Company, California), Martha Wright (retired consultant). Melon: Kevin Crosby, Yan Ren, Bhimu Patil (Fruit & Vegetable Improvement Center, Texas A&M University & Texas AgriLife Research) Cotton: Marianne Arnold & Jenny Clement (Texas A&M University & Texas AgriLife Research). JC is now with the CSIRO, Australia. TARGET AUDIENCES: Cactus: Opuntia ficus indica breeders and industry worldwide. Environmentalists and range specialists for drought resistant forage. Melon: melon breeders and the melon industry PROJECT MODIFICATIONS: Two collaborations, one with K. Hirschi, Baylor College of Medicine, and S. Roux with University of Texas have been dropped due to lack of funding. Melon research on transgenics, will be probably continued on a small scale but is no longer funded Cotton research is not funded and will be shut down as soon as student finishes her manuscripts.
Impacts
Transformation protocols for western shipper cantaloupe and honeydew melons were developed and transgenic melons exhibiting high levels of carotene in the fruit's rind were created. Further work to improve targeting carotene synthesis in the flesh of the fruit will not be perused due to lack of funding.
Publications
- Ren Y, Bang H, Curtis I, Gould JH, Patil S, Crosby KM. 2011. Agrobacterium-mediated transformation and shoot regeneration in elite breeding lines of western shipper cantaloupe and honeydew melons (Cucumis melo L.). Plant Cell Tiss Organ Cult (2012) 108:147-158
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: Collaborative Research, Calcium. Research with the Hirschi Lab, Baylor College of Medicine. Our MS student who was driving this aspect of our collaboration into calcium research, has successfully completed his Thesis PHENOTYPE ANALYSIS OF TOBACCO LINES EXPRESSING A DEREGULATED ARABIDOPSIS CA-ATPASE (ACA2) and graduated in December (2010). Collaborative Research, Apyrase Roux Lab, University of Texas. Apyrase Study. Ectoapyrase, can modulate the levels of extracellular ATP (eATP). Transgenic MicroTom plants have been transferred to the Roux Lab and are under analysis. Collaborative Fruit Improvement Research Opuntia: Conditions favorable for somatic embryogenesis are under exploration. Melon:Regeneration and transformation procedures for TAMUS lines of melons has been developed. Cotton: Light effects on immature embryo development are under study Hormonal applications to developing cotton bolls appear to alter fiber quality. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Sanchez-Lozoya N, Thompson SM, Jean H. Gould J, Shigaki T, Hirschi KD. Abstract published 2009. Altering Ca2+ Partitioning in Transgenic Plants. III Pan American Plant Membrane Biology Workshop. January 13-16, 2010. Puebla Mexico. Ren Y, Haejeen Bang, Jean Gould, Bhimanagouda S. Patil and Kevin M. Crosby. 2011. Shoot regeneration and ploidy variation in tissue culture of honeydew melon (Cucumis melo L. iodorus). American Society Horticulture Science, Annual Conf. Hawaii. Ren Y, Curtis I, Bang H, Park SO,, Gould J, Patil BS, Crosby K. 2010. Establishing an Agrobacterium-mediated transformation system for western-shipper cantaloupe and honey-dew melons (Cucumis melo L.) Texas A&M AgriLife Conference, January 11-15
Publications
- Y. Ren, Haejeen Bang, Ian Curtis, Jean Gould, Bhimanagouda S. Patil1 and Kevin M. Crosby 2011 Establishment of regeneration and A. tumefaciens-mediated transformation system using western shipper cantaloupe and honeydew melons (Cucumis melo L.). Submitted: Plant Cell Tissue & Organ Culture.
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: A. Gossypium hirsutum Texas Marker-1 Early Embryo Development. We have studied the effect of light on viability and successful germination of excised immature (17, 18, 19 dpa) cotton embryos cultured in vitro. This research is part of a PhD Dissertation and manuscript, which has not yet been published. B. Vitis vinifera Tissue Culture & Transformation. We do not have funding for this project and it has been abandoned. C. Opuntia Ficus-indica transformation for different aspects of fruit development. Agrobacterium-based transformation of is used and transgenic plants are being generated and evaluated. This research is funded through a commercial interest. All details and results are proprietary. D. Regulation of Calcium bioavailability in plants. This research is in collaboration with Kendal Hirschi, USDA-ARS Children's Nutrition Center, Baylor College of Medicine. Transgenic carrots, tobacco and Arabidopsis expressing ACA2, a calcium over-expresser gene, have been generated and plants are being analyzed. PARTICIPANTS: Marianne Arnold MEPS TAMU; Claudine Debona, Brazil; Kendal Hirschi USDA-ARS Baylor College of Medicine; Sean Thompson MEPS TAMU. TARGET AUDIENCES: A. Researchers in the area of cotton embryo development and regulation; Cotton breeders interested in wide inter-specific crosses and embryo rescue; B. Abandoned; C. Investigators interested in modifying embryo, seed and fruit development in commercial lines of cactus; D. Researchers interested in increasing calcium nutrition in plants. PROJECT MODIFICATIONS: Vitis project dropped Calcium project added
Impacts
A. Cotton Embryo Development. Our research concerns the hormonal and environmental controls of immature cotton embryo and seed development. The importance of this research is: (1) it will be the first report of the efficacy of light in the development of immature cotton embryos and seed; (2) it will provide a bio-assay for studying the genetic regulation of cotton embryo development, and (3) it will provide a method for the recovery of viable immature cotton embryos (embryo rescue) from wide inter-specific crosses that have become important in improvement of cotton germplasm. B. Vitis Tissue Culture & Transformation. This research is designed to develop a genotype-independent, Agrobacterium-based method of Vitis transformation that will be used for transferring resistance, or tolerance, to Pierce's disease. Pierce's disease has the potential to decimate the production of grapes in the US. C. The impact of our lignin silencing study will be to assist in reducing the lignin content of seeds in some fruit, allowing seeds to be more edible.
Publications
- Sanchez-Lozoya N, Thompson SM, Jean H. Gould J, higaki T, Hirschi KD. Abstract published 2009. Altering Ca2+ Partitioning in Transgenic Plants. III Pan American Plant Membrane Biology Workshop. January 13-16, 2010. Puebla Mexico.
- Ren Y, Crosby K, Bang H, Gould J. 2008. Tissue culture of muskmelon (Cucumis melo L.) ASHS-2008 Annual Conference, 21 - 24 July, Orlando, FL.
- Bona CM, Gould J, Miller JC, Stelly DM, Louzada ES. 2008. Asymmetric hybridization: a tool to be used in Citrus radiation map creation Abs 3107 ASHS-2008 Annual Conference, 21 - 24 July, Orlando, FL.
- Bona CM, Gould J, Miller JC, Stelly DM, Louzada ES. 2008 In vitro regeneration of somatic symmetric and asymmetric hybrid citrus plantlets produced via protoplast fusion Abs 3106 ASHS-2008 Annual Conference, 21 - 24 July, Orlando, FL.
- Bona CM, Gould J, Miller JC, Stelly DM, Louzada ES. 2008. Symmetric and asymmetric somatic hybridization in Citrus spp. for scion genetic diversity generation Abs 3105 ASHS-2008 Annual Conference, 21 - 24 July, Orlando, FL.
- Published Abstracts - Gould J, Arnold M 2009. The effects of light on development of true shoots in excised immature cotton embryos Texas Marker-1 cultured in vitro ASPB annual meeting, July 17-23, Honolulu.
- Gould J, Arnold M. 2008. Evaluation of cultural conditions for immature cotton embryo development into plants using image analysis. ASPB annual meeting. June 22-26, Merida Mexico.
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: Research work is continued with Opunita transformation and analysis. Project is supported by industry; Publication or presentation of this research is not permitted. One proposal regarding tissue culture and transformation of vitis varieties important in Texas for resistance to Pierce's disease, was not funded. Research on early embryo development in cotton is progressing slowly, and is not funded. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Commercial grape producers in Texas PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
None
Publications
- No publications reported this period
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Progress 01/01/07 to 12/31/07
Outputs
OUTPUTS: A. Cotton Embryo Development. During this period, we have found that light improves viability and successful germination of imature (17, 18, 19 dpa) cotton embryos, and we are studying this phenomena. B. Vitis Tissue Culture & Transformation. Shoot apex based transformation is being developed for several anceint clones of Vitis vinifera. C. Specific Regulation of lignin Biosynthesis in Fruit and seed coat. We have been studying the specificity of transient expression using a number of GUS based constructs driven by a number of native plant promoters associated with lignin biosynthesis in developing fruit. After the most specific constructs were found,constructs were placed into binary vectors and into Gateway vectors and used in Agrobacterium-based transformation of Arabidopsis. A population of transgenic plants is being generated and evaluated.
PARTICIPANTS: Marianne Arnold, TAMU Claudine Debona, TAMU B. Gregory Cobb, TAMU G. R. McEachern, TAMU
TARGET AUDIENCES: A. Cotton Breeders interested in wide interspecific crosses and emryo rescue. B. Investigators interested in Vitis improvement, transformation and resistance to Pierce's disease. C. Investigators interested in modifying embryo, seed and fruit developemnt
Impacts
A. Cotton Embryo Development. Our research concerns the hormonal and environmental controls of immature cotton embryo and seed development. The importance of this research is: (1) it will be the first report of the efficacy of light in the development of immature cotton embryos and seed; (2) it will provide a bio-assay for studying the genetic regulation of cotton embryo development, and (3) it will provide a method for the recovery of viable immature cotton embryos (embryo rescue) from wide inter-specific crosses that have become important in improvement of cotton germplasm. B. Vitis Tissue Culture & Transformation. This research is designed to develop a genotype-independent, Agrobacterium-based method of Vitis transformation that will be used for transferring resistance, or tolerance, to Pierce's disease. Pierce's disease has the potential to decimate the production of grapes in the US. C. The impact of our lignin silencing study will be to assist in reducing the
lignin content of seeds in some fruit, allowing seeds to be more edible.
Publications
- Arnold, M., Gould, J. 2007. In vitro conditions for examination of early cotyledonary to maturation stage cotton embryos. Poster #P29019. American Society for Plant Biology.,Chicago Ill.
- Debona, C., Gould, J., Miller, J.C., McEachern, G.R., Setamou, M., Lousada, E. 2007. In vitro manipulation of nine grape cultuvars. Subtropical Plant Sci. 59: In Press.
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Progress 01/01/06 to 12/31/06
Outputs
No progress reported this period
Impacts
Crop improvement
Publications
- Claudine M. Bona1.4, Jean H. Gould2, J. Creighton Miller, Jr.1, George R. McEachern1 and Eliezer S. Louzada3. 2007. In Vitro Manipulation of Nine Grape Cultivars. HortSicence
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Progress 01/01/05 to 12/31/05
Outputs
Funding: We are funded through the TAES Breeder-Driven Molecular Mechanisms for Monocots program through 8/31/2006 to transform elite lines of turfgrass. We are also funded by a contract to transform cactus (proprietary). Transformation of TAES turfgrass lines of bentgrass and zoysia for floral disruption to eliminate pollen production. Putative transgenic plants are being regenerated from Agrobacterium (shoot apex/shoot meristem) and Biolistic procedures. Regenerated plants are under analysis for transferred floral disruption genes. Silencing cottonseed gossypol biosynthesis & accumulation project. In vitro embryo rescue was successfully established for Texas Marker-1 embryos isolated 10 to 20 days after flowering for use in studying gossypol biosynthesis in developing embryos. RNAi-based silencing constructs have been completed. Transient expression studies are in progress.
Impacts
(1.) Elimination of flowering in turfgrass will: (i) reduce production of pollen allergens, (ii) eliminate the capacity for gene flow from transgenic turfgrass to other grasses in the environment, (iii) improve appearance and decrease maintenance of grasses by eliminating flowering stalks and seed heads; (iv) increase biomass. (2.) The impact of silencing gossypol synthesis in the seed of normally glanded cotton cultivars will open new feed and food markets for cottonseed worldwide, by detoxifying the otherwise nutritious cottonseed, a poorly utilized by product of cotton fiber production. The reduction of gossypol in cottonseed will provide increased financial return to cotton growers in that two crops can be generated in the land space and cost of one crop.
Publications
- No publications reported this period
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Progress 01/01/04 to 12/31/04
Outputs
Gossypol Project. (1) Expression of a number of seed specific promoters have been tested via transient expression using biolistcs. (2) A binary vector based on the pKannibal has been constructed containing the best promoter and fragments of RNAi based silencing targetd to the delta-cadinene synthase c1 gene. (3) An in vitro embryo rescue method was developed for cotton ovules (Texas Marker 1) dpa 14-20, for use in gossypol synthesis bioassays. Turfgrass project. (1) Media of shoot proliferation developed. (2)Agrobacterium mediated transformation initiated to knock tou flowering.
Impacts
(1.) The impact of silencing gossypol synthesis in the seed of normally glanded cotton cultivars will open new feed and food markets for cottonseed, providing increased financial return to growers from a cotton crop. (2.) The impact of demonstrating transformation competency of the Russian spring wheat Saratovkaya-29 will open up the improvement of wheat cultivars derived from this popular parent in countries of the former Soviet Union.
Publications
- No publications reported this period
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Progress 01/01/03 to 12/31/03
Outputs
(1.) Putative transgenic cotton plants have been analyzed for low cottonseed gossypol and plants identified. (2.) Real Time RT PCR is being used to study expression of the genes in the gossypol biosynthetic pathway during embryo development. (3.) RNAi-based binary vectors for use in Agrobacterium-mediated transformation using the phaseolin promoter are under development for use in silencing key elements of the gossypol biosynthetic pathway. This step has been problematic. (4.) We have generated putative transgenic plants of a Russian wheat, Saratovkaya-29. Both Agrobacterium and particle bombardment approaches appeared to be effective. The transferred marker genes have been detected by PCR gel blotting followed by hybridization, in regenerated plants and in the progeny of these plants. Genomic DNA analysis will be carried out to determine if transferred genes have been incorporated into the genomes of these plants and inherited.
Impacts
(1.) The impact of silencing gossypol synthesis in the seed of normally glanded cotton cultivars will open new feed and food markets for cottonseed, providing increased financial return to growers from a cotton crop. (2.) The impact of demonstrating transformation competency of the Russian spring wheat Saratovkaya-29 will open up the improvement of wheat cultivars derived from this popular parent in countries of the former Soviet Union.
Publications
- Kopbayev, A. 2004. MS Thesis. Comparison of biolistic and Agrobacterium-mediated genetic transformation of immature and mature embryos of spring wheat cultivar Saratovskaya-29.
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Progress 01/01/02 to 12/31/02
Outputs
The project concerned the development and improvement of transformation & regeneration protocols for the genetic improvement of non-model, elite and recalcitrant plant genotypes of commercial value. In the course of the project, we have developed transformation and regeneration protocols for loblolly pine; transferred a pathogen resistance factor (PR-1) from tobacco into cotton; transferred an antisense construct to block cottonseed gossypol synthesis; compared the mutation rates for callus-based and shoot meristem-based transformation methds in TAES rice; transferred a herbicide resistance gene into winter wheat; and studied the regulation of a drought responsive promoter isolated from loblolly pine. Our research has been focused on facilitating production of transgenic crop plants and forest trees in non-model and elite germplasm, as well as studying the creation of novel genetic traits capable of conferring added value, such as improved, drought tolerance, disease
resistance and reduction of cottonseed gossypol. In the process, we have studied basic plant transformation and regeneration processes. Our current projects concern the genetic silencing of gossypol biosynthesis in cottonseed in the non-model, elite TAES cotton germplasm. Scientists who trained in our lab, have established our 'Shoot Apex' transformation procedure in India, where they have successfully transferred a Bt insecticidal crystal toxin into a non-model Indian cotton cultivar. Prior to this, the regional cotton cultivars of India could not be regenerated following transformation using conventional callus-based, or embryogenesis-based methods
Impacts
The expected results of this research is to facilitate the transformation and regeneration of elite regional genotypes and to make these procedures easily accessible to researchers in academic, extension or governmental research centers. The potential for impact can be seen in the example of the adaptation of this transformation technology by Indian academic center and an Indian seed company, for the transformation of regional cotton cultivars for increased resistance to insects and disease.
Publications
- Wang JT, Gould JH, Padmanabhan V, Newton RJ. 2002. Analysis and localization of the water deficit stress induced gene (lp3). Accepted. Jour. Plant Growth Regulation
- Gould JH, Zhou Y, Padmanabhan V, Magallanes-Cedeno M, Newton RJ. 2002. Transformation and regeneration of loblolly pine: shoot apex inoculation with Agrobacterium. Molecular Breeding 10(3):131-141.
- Gould J, Newton R. 2002 Transformation of Intact Plants Using Agrobacterium' US Patent Application.
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Progress 01/01/01 to 12/31/01
Outputs
Projects funded in 2001 were:'Gossypol genes-transformation and regeneration'funded by Cotton Inc. and 'Antisense Approach to Reduce Gossypol in Cottonseed'funded by the Texas Higher Education Coordinating Board-Advanced Technology Program (2 years). Poster 'IAA & ABA in Developing Seeds & Fibers of Gossypium hirsutum & G. barbadense', was presented at the Annual Meeting of the American Society of Plant Biology,Providence RI.
Impacts
We have a transgenic line of elite TAES cotton that contains the tobacco osmotin gene. These plants are being evaluated for improved resistance to the seedling disease complex and improved drought tolerance. If resistance is signigficant, these plants will become incorporated into the TAES cotton breeding program.
Publications
- 2001 Jau Tay Wang PhD Plant Physiology. In Analysis of Water-Deficit Stress Induced Gene (lp3) May, 2001 TAMU.
- Provisional Patent Application 2001. TAES Inv. Discl. No. 926, Gould & Newton, `Transformation of Intact Plants Using Agrobacterium' Locke, Liddell & Sapp, L.L.P., Houston TX, Dec. 4, 2001, sponsored by Plant Biosciences Ltd.
- Gould JH, Harold Slator, R.H. Smith, Page Morgan. 2001. IAA & ABA in Developing Seeds & Fibers of Gossypium hirsutum & G. barbadense: analyzed by GC-SIM-MS. Proceedings: Cotton Fiber and Seed Quality Conference, San Antonio, published.
- Li H, Luo J, Gould JH. 2001. A Rapid and High Yielding DNA Miniprep for Cotton (Gossypium spp.). Plant Molecular Biology Reporter 19:1-5.
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Progress 01/01/00 to 12/31/00
Outputs
TEX08458 - J. H. Gould Currently funded research projects include the transformation of cotton to improve seed meal quality for livestock feed, transformation of winter wheat to impart herbicide tolerance and development of an automation prototype for plant tissue culture micropropagation. Cotton transformation projects include GOSSYPOL GENES - PLANT TRANSFORMATION & REGENERATION, which focuses on the transformation of TAES cotton varieties using sense and antisense genetic constructs to knock down production of gossypol in cotton seed. Low seed gossypol will allow use of cotton seed in animal feed. Transgenic plants have been recovered and are being analyzed. Transgenic cotton containing new genes for disease resistance and drought tolerance have been developed. WHEAT TRANSFORMATION FOR HERBICIDE RESISTANCE is supported by Americian Cyanamid-BSAF. In this project we are transforming a european variety of winter wheat with Am. Cyanamid's AHAS gene from maize. This
gene confers resistance to IMI herbicides. Transgenic plants have been recovered and brought to seed. Progeny is being analyzed. Our NASA project, AUTOMATION OF CLONAL PROPAGATION, has produced a prototype mechanism for clonal propagation of grass. It is being developed in collaboration with the Mechanical Engineering Dept. at TAMUS. In 2000, we presented aspects of the above research at the ASPP and ISPMB meetings. Luo J, Li H, Hemphill JK, Raisor M, Gould J. 2000. Poster, Transformation of Texas cotton for disease resistance. American Soc. Plant Physiol. Ratnayaka I, Gould J. 2000. Poster, Comparison of Transgenic Rice Plants Produced from Shoot Apex or Callus Inoculation with Agrobacterium. American Soc. Plant Physiol. Ratnayaka I, Gould J. 2000. Poster, A comparison of callus v. shoots in rice transformation by Agrobacterium. Congress, International Society for Plant Molecular Biology. Gould J, Zhou Y, Padmanabhan V, Magallanes-Cedeno M, Newton R. 2000. Poster, Transformation and
regeneration of loblolly pine using shoot apex inoculation with Agrobacterium. Congress, International Society for Plant Molecular Biology, Quebec City, CN. Gould J, Harold Slator, R Smith, Page Morgan. 2000. Invited talk, Free IAA & ABA in Developing Fibers & Embryos of Gossypium hirsutum & G. barbadense: analyzed by GC-SIMS. Cotton Biotechnology Conference, San Antonio. Gould JH. 2000 Crop Biotechnology Update. Invited talk, TAES Entomology Science Conference
Impacts
We have a transgenic line of elite TAES cotton that contains the tobacco osmotin gene. These plants are being evaluated for improved resistance to the seedling disease complex and improved drought tolerance. If resistance is signigficant, these plants will become incorporated into the TAES cotton breeding program.
Publications
- Jinhua Luo. 2000. MS Thesis in Plant Physiology. Transformation of Cotton for Disease Resistance.
- Gould JH, Harold Slator, R.H. Smith, Page Morgan. 2000. Free IAA & ABA in Developing Fibers Embryos of Gossypium hirsutum & G. barbadense: analyzed by GC-SIMS. Proceedings: Cotton Fiber and Seed Quality Conference, San Antonio.
- Luo J, Arriaga J Li H, Hemphill J, Keller N, Isakeit T, Gould J. 2000. Transformation of TAES cotton for seedling disease resistance. Proc: Beltwide Cotton Prod Conf, National Cotton Council.
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Progress 01/01/99 to 12/31/99
Outputs
In 1999, we initiated a new project funded by American Cyanamid to transform wheat for herbicide tolerance. We also continued work in the transformation and improvement of public-sector germplasm. Rice has been transformed using Agrobacterium and both shoot apex and callus based methods and the efficiency of transformation and the occurrence of mutations were compared (Ratnayaka PhD Dissertation TAMU 1999). Our demonstration of stable transformation in loblolly pine (WGTIP improved lines) has been submitted for publication. TAES varieties of Sorghum are under evaluation at the T3 generation, as reduction of gossypol in cotton seed, and increasing disease/stress tolerance by the transfer of resistance genes PR-1 and Osmotin (PR-5). Plants and progeny are transgenic for PR-1. This line will be evaluated for increased resistance to the seeding disease complex. Genetic transformation was achieved using shoot apex inoculation with Agrobacterium (TAMUS US Patent 5,164,310)
which we developed in cotton, and pioneered in the cereals (Gould etal., 1991, Plant Physiol.95:426-434).
Impacts
(N/A)
Publications
- Luo J & Gould J H. 1999. In vitro shoot tip grafting improves recovery of cotton plants from culture. Plant Cell, Tissue & Organ Culture.
- Gould J H, Magallanes-Cedeno M. 1999. Adaptation of Shoot Apex Agrobacterium Inoculation & Culture to Cotton Transformation. Plant Mol. Biol. Rept. 16(3) 284-289.
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Progress 01/01/98 to 12/31/98
Outputs
We worked to demonstrate the feasibility and efficiency of the Shoot Apex plant transformation method with varieties of rice, sorghum, cotton and loblolly pine developed and grown in Texas. Gene transfer was achieved using shoot apex inoculation with Agrobacterium which we developed in cotton, and pioneered in cereals and loblolly pine. We demonstrated stable gene transfer to recently released Texas rice variety, Dixiebelle. We carried out the sorghum transformation project and identified 4 putatively transgenic parent plants using PCR within the population created.A proposal to transform wheat for herbicide resistance was funded. We have produced transgenic cottom plants in improved germplasm.
Impacts
(N/A)
Publications
- Li T., Harris, M., Nessler, C., Gould, J.H., Smith, C.W. 1998. Oviposition, growth and development of Bemesia tabaci (Homoptera: Aleyrodidae) on transgenic tobacco expressing TyDC & TDC. Submitted, J Econ. Ent.
- Gould JH, Zhou Y, Padmanabhan V, Magallanes-Cedeno M, Newton RJ. 1998. Transformation of loblolly pine by shoot apexinoculation with Agrobacterium. Sumbitted, Nature Biotechnology.
- Luo J & Gould JH. 1998. In vitro shoot apex grafting improves recovery of intact cotton (Gossypium hirsutum L.) plants from culture. Submitted, Plant Cell, Tissue & Organ Culture.
- Gould JH, Magallanes-Cedeno M. 1997. Adaptation of Shoot Apex Agrobacterium Inoculation and Culture to Cotton Transformation. Plant Mol. Biol. Rept. 16 (2)
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Progress 01/01/97 to 12/31/97
Outputs
We have been working with rice, sorghum, Texas cotton and loblolly pine to demonstrate the efficiency and stability of gene transfer mediated by shoot apex/Agrobacterium, pioneered in cereals (Gould et al. 1991,Plant Physiol. 95:426-434), when applied to a broad range of commercial plant species. In 1997, we were able to demonstrate stable gene transfer and inheritance in rice using Agrobacterium and we are comparing our shoot apex method with the traditional callus-based method. Although Agrobacterium is now beginning to be used to treansform rice and other cereals, the callus-based, genotype-limited plant regeneration methods are still being used. Aside from genotype restrictions, these methods impose a wide range of culture induced genetic mutations (somaclonal variation) that which are permanent and are carried forward into subsequent generations. Plants derived from both techniques have been determined to be transgenic based on PCR and genomic DNA Southern
hybridization. Shoot derived plants appeared normal, produced a normal yield of fertile seed and progeny. Callus derived plants appeared less robust and exhibited low fertility. The limited progeny set obtained from callus derived plants are still being analyzed and many do not appear to carry the transfered genes. Many of the 3rd generation progeny (R3) of shoot derived plants appear to carry the genes in high molecular weight DNA based on Southern DNA hybridization to HindIII digested DNA, characteristic of genetic transformation. Overall, the shoot-based peocedure was easiest, plants were recovered faster (4 weeks), plants appeared normal and were fertile, and the transferred genes can be detected in the progeny. On the other hand, callus derived plants required development of a regeneration medium, 2-3 months of culture, regenerated plants did not appear normal nor did they exhibit normal fertility (Data presented at Plant & Animal Genome VI, January 1998).
Impacts
(N/A)
Publications
- Zhou, Y., Newton, R., Gould, J. 1997. A simple method for identifying plant/T-DNA Junction Sequences Resulting from Agrobacterium-mediated DNA Transformation. Plant Mol Biol. Reporter, 15:246-254.
- Gould, J.H. 1997. Transformation of the Cereals using Agrobacterium. In: RS Tuan (Ed), Methods in Molecular Biology, Humana Press Inc., Totawa,NJ. Vol. 62:489-499.
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