Project Number: 6208-21000-016-05
Project Type: Reimbursable

Start Date: Jan 01, 2008
End Date: Dec 31, 2008

Objective:
The research proposed on cotton fiber specific promoter (CFSP) study for 2008 will focus on: (1) Advance the 35S::GFP, CFSP::GFP and Fb::GUS transgenic lines to T3 stage and identify the homozygous transgenic plants for each line; (2) Large seed increase in the field for homozygous transgenic lines of CFSP::GUS and corresponding controls; (3) Evaluate fiber gene expression pattern of transgenic plants under normal growth conditions; and (4) Evaluate the impact of environmental stresses (drought, high temperature (HT) and/or drought and HT) on fiber gene expression, fiber production and fiber quality.

Approach:
(1) Grow 3 T1 plants per line, 6-8 independent transgenic lines per construct to produce T2 seeds. The transgene activity in T1 plant will be examined at proper fiber developmental stage either by GUS staining or GFP florescence. Eight to ten T2 plants from selected transgenic lines will advance to T3 generation. The transgene activity in T2 plant will also be examined. (2) Grow out all available CFSP T3 homozygous and null transgenic lines in the field in year of 2008 to produce enough T4 seeds which will serve as a central resource for cotton research community. Transgene activity of these plants will be examined during the growing season. (3) We evaluate gene expression pattern of those transgenes in each of the transgenic line under normal growth conditions. Cotton bolls at various fiber developmental stages will be harvested. Total proteins from the fiber tissue will be extracted and GUS activities will be examined quantitatively. If necessary, quantitative RT-PCR will be performed using mRNA as template to quantify the level of gene expression. (4) The impact of environmental stresses on fiber gene expression (such as moderate and severe drought, high temperature or combination of both HT and moderate drought) will be evaluated in green house or growth chambers. Drought stresses will be applied either for 7-10 days at different fiber developmental stages or throughout the entire fiber developmental stages. Levels of fiber gene expression will be examined the same way as in Plan #3. Fiber and seed production per boll and fiber quality will also be evaluated. The impact of stress applied on all measured parameters will be analyzed and compared with those of controls. Three quarterly and one annual report will be provided to Cotton Incorporated in a timely manner. Research will also be published in scientific journals. IBC RECERTIFICATION SEPTEMBER 2, 2008.