Progress 09/01/04 to 08/31/09
Outputs
OUTPUTS: The specific objectives of this project are to refine in vitro plant regeneration methods amenable to gene delivery for select guava cultivars, standardize protocols for Agrobacterium-mediated genetic transformation of popular guava genotypes using CBF cold hardiness genes, and regenerate and evaluate transgenic guava plants for cold tolerance in the greenhouse and field trials. To accomplish these objectives, the in vitro plant regeneration protocols adopted during guava research project of 2000, were examined and adopted to improve their reproducibility using organogenesis and somatic embryogenesis protocols along with their feasibility for genetic transformation using Agrobacterium and PDS-1000 biolistic gun. Crucial changes were made in culture media supplements and ambient culture conditions to achieve desired results. In 2009, we investigated multiple shoot regeneration and in vitro proliferation of Lucknow-49 and Gushiken Sweet guava cultivars in response to plant growth regulators (PGR). Effects of PGR and genotype on nodal explants we studied by supplementing MS medium with several concentrations and combinations of benzyl adenine, kinetin, adenine hemi-sulphate and thidiazuron. Moreover, feasibility evaluation of promising in vitro procedures for genetic transformation was attempted and we further examined these steps during the remaining period in 2009. The plant regeneration protocols using organogenesis and somatic embryogenesis were attempted to regenerate additional guava tissues and cultivars that produce superb quality of healthy fruits. Using gene transfer protocols, we attempted and evaluated putative transgenic guava plantlets for cold tolerance in vitro and also in the specialty plants house where they were exposed to cold more than their comparable plants. A graduate student placed through this project in Horticulture Department at the Iowa State University for a PhD degree, later decided to complete her MS Degree in 2008 and moved to another institution on the east coast for her PhD education. While at Iowa, she engaged in her thesis research using physiological studies on four guava cultivars including Beaumont, Ruby x Supreme, Red Fleshed, and Lucknow-49. After completing her required courses, she intensified her physiological research efforts during 2008 and obtained encouraging results. Then in late Fall she completed her thesis and defended in late November 2008. We also received leaf samples of 24 guava cultivars from Hawaii and extracted DNA for microsatellite work. To determine antioxidant activity, we analyzed white, cream and red flesh colored guava fruits for total phenolics. Distinct varietal disparity was observed among cultivars for these chemicals. Genetic structure of some guava (Psidium guajava) accessions available in the US was determined using microsatellite markers during 2009. In addition, the level and distribution of variation among these accessions were assessed. Fourteen primers were tested in the 2009 study which generated amplicons and detected polymorphisms among the guava cultivars. The microsatellite research is continued during 2010 although the project duration has been terminated. PARTICIPANTS: In addition to working on this project, the PD/PI, Co-PD/PIs as well as research professionals also participated in scientific conferences and presented research findings on guava and other specialty plants research. A graduate student for the Doctor's Degree placed at the Iowa State University through support from this project, decided instead to complete MS and joined another institution. We employed several undergraduates and also mentored K-12 school students involving them in guava research. Furthermore, we developed/strengthened partnership linkages and collaborative ventures with the USDA-ARS, the Iowa State University, Clemson University, and NASA Plant Science Research Group at Kennedy Space Center. As we planned for our future initiatives as related to beyond the original duration of this project; we developed a couple of new project proposals involving antioxidants and other nutraceutical ingredients of guava fruit and submitted to the USDA NRI in 2009 and to CBG Grants Proposal during FY2010. TARGET AUDIENCES: We consider that the American consumer and particularly the specialty plants growers in the mild temperate region of the United States and especially in the southeastern states have been our target audiences. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The National Priority for NIFA - Global Food Security and Hunger has been addressed by this research project. Conducting extensive research specifically on the physiology of cold hardiness in guava while developing suitable biotechnology for in vitro plant regeneration protocols that will be amenable to genetic transformation of guava germplasm will facilitate scientists to integrate cold hardiness genes into superior guava genotypes with high quality fruit characteristics. With these research accomplishments, we will be in a position to enhance cold hardiness of guava germplasm and genetically improve the desirable genotypes for adaptation to at least in the mild temperate locations of the USA, like Georgia and the rest of the Southeast. Thus, biotechnology research will assist us to enhance hardiness and further improvement in nutraceutical quality of guava fruit. With the nutraceutical superiority of guava fruit as evident from our bioactivity evaluation over majority of other fruits, its successful production will benefit the southeastern growers as well as the consumers by providing them wealth and health benefitting fruit, respectively.
Publications
- Viji, G., Harris, D. L., and Yadav, A. K. 2009. Use of microsatellite markers in the characterization of genetic diversity of guava (Psidium guajava L.) in the USA. Molecular Markers in Horticulture Symposium, Corvallis, OR, July 29 - August 1, 2009.
- Viji, G., Harris, D. L., Zee, F. T. P., and Yadav, A. K. 2009. Use of microsatellite markers to characterize genetic diversity of selected accessions of guava (Psidium guajava L.) in the US (accepted - Acta Horticulturae, International Society for Horticultural Science).
- Viji, G., Harris, D.L., and Yadav, A. K. 2009. Multiple shoot induction and proliferation from nodal explants of guava in vitro. 15th Biennial Research Symposium, March 28-April 1, 2009. Atlanta, GA.
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: The specific objectives of this project are to further refine in vitro plant regeneration protocols amenable to gene delivery techniques for selected guava cultivars, standardize protocols for Agrobacterium-mediated genetic transformation of desirable guava genotypes using CBF series of cold hardiness genes, and regenerate and evaluate transgenic guava plants for cold tolerance in the greenhouse and/or field trials. In order to accomplish these objectives, the in vitro plant regeneration protocols developed / refined during earlier biotechnology research project on guava were examined and/or advanced to improve their reproducibility by means of organogenesis and somatic embryogenesis protocols along with their feasibility for genetic transformation using Agrobacterium and PDS-1000 biolistic gun. Various changes were made in the culture media supplements and ambient conditions to achieve desired results. Furthermore, feasibility evaluation of promising in vitro procedure(s) for genetic transformation was attempted; we will continue to further examine these steps during the remaining time in 2009. The plant regeneration protocols using organogenesis and somatic embryogenesis will be attempted to regenerate additional guava tissues and/or cultivars that produce an excellent quality fruits. For gene transfer protocols, we will keep up with our attempts. Putative transgenic guava plantlets will be evaluated for cold tolerance in vitro and also in the specialty plants house as well as in the physiological studies. Our graduate student placed through this project at the Iowa State University for Ph.D. degree, decided to complete an MS degree in 2008 and move on to another institution for Ph.D. She engaged in thesis research using physiological studies on 4 guava (Psidium guajava L.) varieties (Beaumont, Ruby x Supreme, Red Fleshed, and Lucknow-49). Following completion of required courses, she intensified her physiological research efforts during 2008 and obtained encouraging results. Then in late Fall she completed her thesis and defended in late November 2008. We received leaf samples of 24 guava cultivars from Hawaii and extracted DNA for microsatellite data. In order to determine antioxidant activity, we got analyzed white, cream and red flesh colored guava fruits for total phenolics. Distinct varietal variation was obvious. PARTICIPANTS: In addition to working on the project, the PD/PI, Co-PD/PI, and research professional also participated in scientific conferences and presented research findings. A graduate student for the Doctor's Degree placed at the Iowa State University through support from this project, decided instead to complete MS and joined another institution. We employed several undergraduates and also mentored K-12 school students involving them in guava research. Furthermore, we developed/strengthened partnership linkages and collaborative ventures with the USDA-ARS, the Iowa State University, Clemson University, and NASA Plant Science Research Group. We have planned future initiatives as related to beyond the duration of this project; we have already initiated development of a new project plan involving antioxidants and other nutraceutical ingredients of guava fruit for submitting to either the USDA NRI or CBG Grants Proposal during FY2009. TARGET AUDIENCES: The specialty plants growers in the mild temperate region of the United States, and especially in the southeastern states. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Conducting more specific research on the physiology of cold hardiness in guava while developing suitable biotechnology for the in vitro plant regeneration protocols amenable to genetic transformation of guava will facilitate scientists to integrate cold hardiness genes into superior guava varieties with quality fruit. With these accomplishments, we will be in a position to enhance cold hardiness of guava germplasm and genetically improve them for adaptation to at least in the mild temperate areas of the USA like Georgia and the rest of the Southeast. With the nutraceutical superiority of guava as evident from our bioactivity tests over majority of fruits, its successful production will benefit the southeastern growers.
Publications
- Joshee, N., Rimando, A. M., Parajuli, P., Rawat, G. S., and Yadav, A. K. 2009. Investigating two medicinal Scutellaria species of Himalayan origin. Pp. 347-356, In Singh S. B., Chaurasia, O. P., Yadav, A., Rimando, A. M., and Terrill, T. H. (Eds.), Advances in Agriculture, Environment, and Health; Fruits, Vegetables, Animals and Biomedical Sciences, S S Publishing House, Delhi, India.
- P. Parajuli, N. Joshee, A. Rimando, S. Mittal and A. K. Yadav. 2009. In vitro anti-tumor mechanisms of various Scutellaria extracts and constituent flavonoids. Planta Medica 75:41-48.
- A. K. Yadav, G. Viji, N. Joshee, B. K. Biswas, E. A. Baldwin, and F. T. P. Zee. 2008. Guava Biotechnology: Road toward enhancing plant cold hardiness and nutraceutical values of fruit. 2nd International Symposium on Guava and Other Myrtaceae, Merida, Mexico, November 9-13, 2008, Abstracts Books 53-54.
- Hao, W., R. Arora, A. K. Yadav, and N. Joshee. 2008. Freezing tolerance and cold acclimation in Guava (Psidium guajava L.). HortScience (Accepted and in press)
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Progress 01/01/07 to 12/31/07
Outputs
OUTPUTS: The objectives of this project are to further refine in vitro plant regeneration protocols amenable to gene delivery techniques for selected guava cultivars, standardize protocols for Agrobacterium-mediated genetic transformation of desirable guava genotypes using CBF series of cold hardiness genes, and regenerate and evaluate transgenic guava plants for cold tolerance in the greenhouse and/or field trials. Thus, to accomplish these objectives, the in vitro plant regeneration protocols that were developed and/or refined during the earlier guava biotechnology research project were further examined and/or advanced for their reproducibility using organogenesis and somatic embryogenesis protocols along with their feasibility for genetic transformation using Agrobacterium and biolistic gene gun. Necessary changes were made in the tissue culture medium and/or culture conditions for achieving approximate results. Furthermore, feasibility evaluation of promising in vitro
procedure(s) for genetic transformation was also attempted; we will continue to further examine it during the 2008 season. The protocols for both the organogenesis and the somatic embryogenesis will be attempted to regenerate additional guava cultivars capable of producing excellent quality fruits. For accomplishing genetic transformation, we will continue attempting to co-cultivate embryogenic tissues from guava zygotic embryos, with Agrobacterium during the 2008 fruiting season (April-June) which seems to be the most suitable time of the year when adequate supply of immature zygotic embryos is available to produce adequate embryonic masses. Putative transgenic guava plantlets will be evaluated for cold tolerance in vitro and also in the specialty plants house as well as in physiological studies. Our graduate student placed through this project at the Iowa State University for Ph.D. degree, is engaged in thesis research using physiological studies on four guava (Psidium guajava L.)
varieties (Beaumont, Ruby x Supreme, Red Fleshed, and Lucknow-49). Following completion of required courses, the graduate student will intensify her physiological research efforts during 2008 to collect additional information on some of the encouraging results.
PARTICIPANTS: The PD/PI and Co-PD/PI in addition working on the project also participated in scientific conferences and presented research findings. A graduate student for the Doctor's Degree at the Iowa State University is being supported through this project. We employed several undergraduates and also mentored K-12 school students involving them in guava research. Furthermore, we also developed/strengthened partnership linkages and collaborative ventures with the USDA-ARS-NPURU, the Iowa State University, Clemson University, and NASA Plant Science Research Group. We have planned future initiatives as related to beyond the duration of this project; we have already initiated development of a new project plan involving antioxidants and other nutraceutical ingredients of guava fruit for submitting to either the USDA NRI or CBG Grants Proposal during FY2009.
TARGET AUDIENCES: The specialty plants growers in the mild temperate region of the United States, and especially in the southeastern states.
Impacts
Researching more specifically on the physiology of cold hardiness in guava while developing suitable biotechnology for the in vitro plant regeneration protocols amenable to genetic transformation of guava will facilitate scientists to integrate cold hardiness genes into popular guava varieties. With these accomplishments, we will be in a position to enhance cold hardiness of guava germplasm to genetically improve it for adaptation to at least in the mild temperate areas of the USA such as Georgia and the rest of the Southeast. With the nutraceutical superiority of guava over majority of other fruits, its successful production will benefit the southeastern growers.
Publications
- Biswas, B. K., N. Joshee, A. Yadav, and A. K. Yadav. 2007. Development and Application of Biotechnology in Guava: A Nutraceutical Fruit. Acta Horticulturae (ISHS) 744:267-276.
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Progress 01/01/06 to 12/31/06
Outputs
The objectives of this project are to further refine in vitro plant regeneration protocols amenable to gene delivery techniques for selected guava cultivars, standardize protocols for Agrobacterium-mediated genetic transformation of desirable guava genotypes using CBF series of cold hardiness genes, and regenerate and evaluate transgenic guava plants for cold tolerance in the greenhouse and/or field trials. Thus, to accomplish these objectives, the in vitro plant regeneration protocols that were developed and/or refined during the earlier guava biotechnology research project were further examined and/or advanced for reproducibility using organogenesis and somatic embryogenesis along with their feasibility for genetic transformation using Agrobacterium and biolistic gene gun. Necessary changes were made in the tissue culture medium / conditions for achieving approximate results. Furthermore, feasibility evaluation of promising in vitro procedure for genetic
transformation was also attempted, we will continue to further examine during the 2007 season. The protocols for both the organogenesis and the somatic embryogenesis will be tried to regenerate additional guava cultivars produce excellent quality fruits. For accomplishing genetic transformation, we will continue attempting to co-cultivate embryogenic tissues from guava zygotic embryos, with Agrobacterium during the 2007 fruiting season (April-June) which seems to be the most suitable time of the year when adequate supply of immature zygotic embryos is available to produce adequate embryonic masses. Putative transgenic guava plantlets will be evaluated for cold tolerance in vitro and also in the specialty plants house.
Impacts
Developing suitable biotechnology for in vitro plant regeneration and genetic transformation of guava will enable scientists to integrate cold hardiness genes into popular guava varieties. With that, this will enhance cold hardiness of guava germplasm to genetically improve it for adaptation to at least the mild temperate climatic areas of the United States such as Georgia and the rest of the southeastern USA.
Publications
- Yadav, A K., N. Joshee, and A Yadav. 2007. Innovations and Advances in Medicinal and Nutraceutical Plant Research at the Fort Valley State University. Abstract submitted for the International Symposium on Medicinal and Nutraceutical Plants, March 19-23.
- Biswas, B. K., A Yadav, and A K. Yadav. 2006. Somatic Embryogenesis and High Frequency Plantlet Recovery in Guava. 14th ARD Biennial Research Symposium, Atlanta, Georgia, USA, April 1-5, 2006, p 291.
- Yadav, A. K., N. Joshee, B. K. Biswas, and A Yadav. 2006. Biotechnological Advances in Medicinal and Nutraceutical Plants. Society for In Vitro Biology Annual Conference, Minneapolis, MN, USA, June 3-7. P 3006.
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Progress 01/01/05 to 12/31/05
Outputs
The objectives of this project are to further refine in vitro plant regeneration protocols amenable to gene delivery techniques for selected guava cultivars, standardize protocols for Agrobacterium-mediated genetic transformation of desirable guava genotypes using cold hardiness genes, and regenerate and evaluate transgenic guava plants for cold tolerance in the greenhouse and/or field trials. Thus, to accomplish these objectives, the in vitro plant regeneration protocols that were developed and/or standardized during the earlier guava biotechnology research project were further examined and/or advanced for reproducibility using organogenesis and somatic embryogenesis along with their feasibility for genetic transformation using Agrobacterium and biolistic gene gun. Necessary changes were made in the culture medium / conditions for achieving approximate results. Furthermore, feasibility evaluation of promising in vitro procedure for genetic transformation was also
attempted, it will be further examined during the 2006 season. The protocols for both the organogenesis and the somatic embryogenesis will be tried to regenerate additional guava cultivars with excellent quality fruit. For accomplishing genetic transformation, we will attempt to co-cultivate embryogenic tissues from guava zygotic embryos, with Agrobacterium during the 2006 fruiting season which is the most suitable time of the year when adequate supply of immature zygotic embryos will be available to produce adequate embryonic masses. Putative transgenic guava plantlets will be evaluated for cold tolerance in vitro and in the specialty plants house.
Impacts
Developing suitable biotechnology for in vitro plant regeneration and genetic transformation of guava will enable scientists to integrate cold hardiness genes into popular guava varieties. With that, this will enhance cold hardiness of guava germplasm to genetically improve it for adaptation to at least the mild temperate climatic areas of the United States such as Georgia and the rest of the southeastern USA.
Publications
- Biswas, B. K., A. Yadav, N. Joshee, N. and Anand K. Yadav. 2005. Plant Regeneration in Guava Through Somatic Embryogenesis. In Vitro 41: 26A, P-1002.
- Yadav, A. K., B. K. Biswas, N. Joshee, and A. Yadav. 2005. Development and Application of Biotechnology in Guava: A Nutraceutical Fruit. FAV Health 2005, Intl Symposium on Human Health effects of Fruits and Vegetables, p. 35-36, August 17-20, Quebec City, Canada.
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