Progress 09/01/09 to 08/31/12
Outputs
OUTPUTS: The project objectives addressed for Year I are described: 1) Test optimal regulation of the HRD and SHN genes for plant performance (1-12 months): To test for optimal regulation required for enabling WUE phenotypes, constructs were made for expression in rice of glucocorticoid receptor (GR) fusions to SHN and HRD genes. The HRD-GR fusion is being transformed into Nipponbare. 2) Transform rice orthologs of HRD and SHN genes into adapted rice cultivars (1-12 months): Constructs of the Arabidopsis and rice HRD and SHN genes, named AtHRD, OsHRD, AtSHN, OsSHN were made. Previously the AtHRD and AtSHN constructs were transformed into test cultivar Nipponbare, and were analyzed. The OsHRD and OsSHN constructs under control of the CaMV35S promoter were constructed. The constructs were transformed into test cultivar Nipponbare and the adapted cultivars IR64 (indica) adapted to grow in most parts of Asia, and the US japonica cultivar Cocodrie. Results of Transformation rice cultivars with OsSHN and OsHRD constructs. Construct OsSHN: 11 transgenic lines in Cocodrie; 9 lines in Nipponbare, regeneraants selected in IR64. Construct OsHRD: 38 transgenic lines in Cocodrie; 11 lines in Nipponbare; 3 lines in IR64 have been selected and used for generating seed. 3. Introgress Nipponbare HRD/SHN transgenes into diverse rice cultivars (1-24 months), from different growing regions in the US and Asia: This part of project will not be continued (see Modification below), as IRRI does not have the capacity at present to do this. However, we will be able to achieve the same results by transforming the indica cultivar IR64 under Objective 2, which is on schedule. PARTICIPANTS: PD Andy Pereira (VBI, Virginia Tech); Research Associate Sarah Misyak (VBI, Virginia Tech); Co-PD Dr Niranjan Baisakh (LSU-Agcenter). Collaborations: Our work on developing genotypes for improved WUE, have enabled us to initiate discussion on future collaborations with the Dale Bumpers College of Agricultural, Food and Life Sciences at the University of Arkansas. In addition, within Virginia Tech, the rice gene constructs we have generated are being used by Bingyu Zhao (Horticulture) to transform switchgrass as a C4 grass, to test the WUE enhancements for C4 plants. In addition, we provided rice gene constructs to Jai Rohila (S. Dakota State) to transform into wheat and check for WUE phenotypes as a collaborative project. Training: Postdoctoral Fellow: Madana Ambavaram; contributed to making constructs and analysis of transgenic plants for drought stress response; Technical Personnel: Sarah Misyak, trained in drought physiology and molecular genetics, worked on screening for gene expression and drought physiology of rice genotypes. Scientist visitor from Iraq: Shatha Yousif; was trained in drought physiology and helped with rice transformation in the lab, by the project personnel TARGET AUDIENCES: Drought and rice research scientists, were addressed by 2 talks by PD at the Plant and Animal Genome meeting 2010 in San Diego. PROJECT MODIFICATIONS: The major project modification is that the PD will move to University of Arkansas, at the Department of Crop, Soil and Environmental Sciences, from Jan 2011. A request is made to move this project with the PD to University of Arkansas. Since 2010, Co-PD Subudhi is replaced by Co-PD Baisakh, which has been notified to USDA-NIFA. Participant IRRI will not participate in project due to change in research direction of Co-PD involved. The Objective 3 under IRRI scope of work, will not be possible as IRRI does not have the capacity at present to do this task. The program has now been changed as follows. Objective 3. Introgress Nipponbare HRD/SHN transgenes into diverse rice cultivars (1-24 months), from different growing regions in the US and Asia. We propose to achieve the same results by transforming the indica cultivar IR64 under Objective 2, which is on schedule. We request that the portion of IRRI budget be allocated to PD to complete this objective.
Impacts
Segregation analysis confirmed single gene inheritance for T1 plants from 7 Cocodrie_OsHRD and 4 Cocodrie_OsSHN independent transgenic lines that are being grown to isolate homozygous lines for subsequent drought phenotyping. Analysis of Nipponbare_OsSHN lines displayed increased Water Use Efficiency (WUE) using gravimetric methods. Analysis of gas exchange parameters will be done using LI-COR 6400 to study photosynthesis and stomatal conductance. Collaborations: Our work on developing genotypes for improved WUE, have enabled us to initiate discussion on future collaborations with USDA scientists Brian Scheffler and collaborators at University of Arkansas. In addition, within Virginia Tech, the rice gene constructs we have generated are being used by Bingyu Zhao (Horticulture) to transform switchgrass as a C4 grass, to test the WUE enhancements for C4 plants. In addition, we provided rice gene constructs to Jai Rohila (S. Dakota State) to transform into wheat and check for WUE phenotypes as a collaborative project.
Publications
- Journal Articles Ambavaram MM, Krishnan A, Trijatmiko KR, Pereira A (2010) Coordinated activation of cellulose and repression of lignin biosynthesis pathways in rice. Plant Physiol. online 10.1104/pp.110.168641, PMID: 21205614.
- Book Publication Batlang U, Baisakh N, Ambavaram MMR, Pereira A (2011) Screening for Drought and Salinity Stress Response Phenotypes in Rice. In: Rice Methods, Ed. Yinong Yang, Humana Press, (in press)
- Meeting Abstracts Krishnan A, Harb A, Ambavaram MMR, Batlang U, Loganathan A, Crasta O, Lin L, Wittich P, Pereira A (2010) Systems biology of drought stress response in plants. W290: Genomics of Plant Development and Signal Networks. Plant and Animal Genomes XVIII, Jan 9-13, 2010, San Diego. Harb A, Dixit S, Karaba A, Aharoni A, Krishnan A, Ambavaram MMR, Batlang U, Pereira A (2010) Abiotic stress tolerance and altered root phenotypes. W531: Root Genomics. Plant and Animal Genomes XVIII, Jan 9-13, 2010, San Diego
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Progress 09/01/09 to 08/31/10
Outputs
OUTPUTS: The project objectives addressed for Year I are described: 1) Test optimal regulation of the HRD and SHN genes for plant performance (1-12 months): To test for optimal regulation required for enabling WUE phenotypes, constructs were made for expression in rice of glucocorticoid receptor (GR) fusions to SHN and HRD genes. The HRD-GR fusion is being transformed into Nipponbare. 2) Transform rice orthologs of HRD and SHN genes into adapted rice cultivars (1-12 months): Constructs of the Arabidopsis and rice HRD and SHN genes, named AtHRD, OsHRD, AtSHN, OsSHN were made. Previously the AtHRD and AtSHN constructs were transformed into test cultivar Nipponbare, and were analyzed. The OsHRD and OsSHN constructs under control of the CaMV35S promoter were constructed. The constructs were transformed into test cultivar Nipponbare and the adapted cultivars IR64 (indica) adapted to grow in most parts of Asia, and the US japonica cultivar Cocodrie. Results of Transformation rice cultivars with OsSHN and OsHRD constructs. Construct OsSHN: 11 transgenic lines in Cocodrie; 9 lines in Nipponbare, regeneraants selected in IR64. Construct OsHRD: 38 transgenic lines in Cocodrie; 11 lines in Nipponbare; 3 lines in IR64 have been selected and used for generating seed. 3. Introgress Nipponbare HRD/SHN transgenes into diverse rice cultivars (1-24 months), from different growing regions in the US and Asia: This part of project will not be continued (see Modification below), as IRRI does not have the capacity at present to do this. However, we will be able to achieve the same results by transforming the indica cultivar IR64 under Objective 2, which is on schedule. PARTICIPANTS: PD Andy Pereira (VBI, Birginia Tech) Research Associate Sarah Misyak (VBI, Virginia Tech) Co-PD Dr Niranjan Baisakh (LSU-Agcenter) Collaborations: Our work on developing genotypes for improved WUE, have enabled us to initiate discussion on future collaborations with USDA scientists Brian Scheffler and collaborators at University of Arkansas. In addition, within Virginia Tech, the rice gene constructs we have generated are being used by Bingyu Zhao (Horticulture) to transform switchgrass as a C4 grass, to test the WUE enhancements for C4 plants. In addition, we provided rice gene constructs to Jai Rohila (S. Dakota State) to transform into wheat and check for WUE phenotypes as a collaborative project. Training: Postdoctoral Fellow: Madana Ambavaram; contributed to making constructs and analysis of transgenic plants for drought stress response Technical Personnel: Sarah Misyak; trained in drought physiology and molecular genetics. Screening for gene expression and drought physiology of rice genotypes. Scientist visitor from Iraq: Shatha Yousif; was trained in drought physiology and helped with rice transformation in the lab, by the project personnel TARGET AUDIENCES: Drought and rice research scientists, were addressed by 2 talks by PD at the Plant and Animal Genome meeting 2010 in San Diego. PROJECT MODIFICATIONS: Co-PD Dr P.K. Subudhi has been changed by LSU to Dr Niranjan Baisakh. Participant IRRI will not participate in project due to change in research direction of Co-PD involved. The Objective 3 under IRRI scope of work, will not be possible as IRRI does not have the capacity at present to do this task. The program has now been changed as follows. Objective 3. Introgress Nipponbare HRD/SHN transgenes into diverse rice cultivars (1-24 months), from different growing regions in the US and Asia. We propose to achieve the same results by transforming the indica cultivar IR64 under Objective 2, which is on schedule. We request that the portion of IRRI budget be allocated to PD to complete this objective.
Impacts
Segregation analysis confirmed single gene inheritance for T1 plants from 7 Cocodrie_OsHRD and 4 Cocodrie_OsSHN independent transgenic lines that are being grown to isolate homozygous lines for subsequent drought phenotyping. Analysis of Nipponbare_OsSHN lines displayed increased Water Use Efficiency (WUE) using gravimetric methods. Analysis of gas exchange parameters will be done using LI-COR 6400 to study photosynthesis and stomatal conductance. Collaborations: Our work on developing genotypes for improved WUE, have enabled us to initiate discussion on future collaborations with USDA scientists Brian Scheffler and collaborators at University of Arkansas. In addition, within Virginia Tech, the rice gene constructs we have generated are being used by Bingyu Zhao (Horticulture) to transform switchgrass as a C4 grass, to test the WUE enhancements for C4 plants. In addition, we provided rice gene constructs to Jai Rohila (S. Dakota State) to transform into wheat and check for WUE phenotypes as a collaborative project.
Publications
- Harb A, Dixit S, Karaba A, Aharoni A, Krishnan A, Ambavaram MMR, Batlang U, Pereira A (2010) Abiotic stress tolerance and altered root phenotypes. W531: Root Genomics. Plant and Animal Genomes XVIII, Jan 9-13, 2010, San Diego
- Book Publication Batlang U, Baisakh N, Ambavaram MMR, Pereira A (2010) Screening for Drought and Salinity Stress Response Phenotypes in Rice. In: Rice Methods, Ed. Yinong Yang, Humana Press, in press
- Meeting Abstracts Krishnan A, Harb A, Ambavaram MMR, Batlang U, Loganathan A, Crasta O, Lin L, Wittich P, Pereira A (2010) Systems biology of drought stress response in plants. W290: Genomics of Plant Development and Signal Networks. Plant and Animal Genomes XVIII, Jan 9-13, 2010, San Diego.
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