Development Of Transgenic Lines Involved In Two Zinc-Finger Proteins Related To Stresses In Cotton(Gossypiun Hirsutm L.)

Cotton is one of the most important cash crops in the world. As the major cultivar of cotton, upland cotton produces more than90%of the total fiber in the worldwide. With the deterioration of the environment, in cotton growth area, most cotton cultivars are affected by a series of natural disasters, such as drought, salt, low temperature and other stresses, which impact critically the growth and decreasing the yield of cotton. Breeding new cotton varieties with multi-resistances is one of the important tasks of cotton breeding.Zinc finger proteins are a class of proteins with zinc ions specific binding and folded into a finger-like structure. The domain combines zinc ions by cysteine and histidine. The zinc finger domain can combine with DNA, RNA, protein and lipid, Zinc finger proteins are involved in various processes of all life activities in plant, with some regulatory functions in biotic and stress responses.Agrobacterium-mediated genetic transformation in cotton is an effective method with simple, relatively high transformation efficiency, low copy numbers of transferred gene, stable and genetically renewable offspring. Agrobacterium-mediated genetic transformation in cotton has been widely used in the creation of transgenic cotton materials. In the process, the procedure of Agrobacterium-mediated transformation has also been further optimized.In this study, we re-established tissue culture transformation system in cotton by referring to the previous study, and compared the influence of different vectors transformed in the regeneration process of cotton tissue culture. We constructed the plant binary expression vectors of GhZFP2and GhRCHY1, transferred the two zinc finger gene into cotton by Agrobacterium-mediated genetic transformation method, and got To generation of transgenic cotton lines. The main results were as following:We successfully re-established genetic transformation regeneration system of tissue culture in cotton, and recorded morphological characteristics on each growth stage in the process of cotton tissue culture; By means of the tissue culture technology, WO accession, the receptor for cotton genetic transformation regeneration, has been further screened to produce the new regenerating clone plants for improving the high embryo rate and vitality during tissure culture; By comparing the regeneration process with different vectors, we found the receptor with Agrobacterium infection or not, the callus at each growth stage have a great impact, at the same time, the regeneration process was also associated with the function of the target genes transferred. Further, we selected GhZFP2and GhRCHYl related to stress response and analyzed their expression characteristics under different stress induction by real time Q-PCR analysis, the result showed that the expression of GhZFP2was up-regulated significantly after ABA and PEG induction, and the expression of GhRCHY1was up-regulated significantly after ABA, PEG and NaCl treatment; nine different clone lines with pBI121-GhZFP2vector and nine different clone lines with pBI121-GhRCHYl vector were individually obtained via Agrobacterium-mediated genetic transformation regeneration system of tissue culture in cotton. The study will provide transgenic resources with stress-resistance and lay a foundation in the future cotton stress-resistant breeding.