Expression Of Rice HD-ZIP Transcription Factor Oshox4, Oshox6and IPT Gene In The Rice And Drought Resistance Analyse Of These Transgenic Plants

Rice is one of the world's most important food crops, various of biological stress such as drought,cold, high temperature and salinity is significantly impact on plant growth and development. Resulting isdecline yield. Drought is general form of biological stress, and already becomes a big problem foragricultural developing in many places. rice is major crops in our country and it needs huge water duringthe life cycle.so to studying drought tolerance of rice, reducing the water for agricultural production,improve the drought tolerance of rice and cultivate drought resistance varieties is particularly importantIn this study, to obtain a gene that enhanced rice drought genetic improvement as purpose,through overstressing of transcription factor and association with drought tolerance gene in rice, and ingreen house and field systematically analyze for the transgenic plant.Initially identified transcriptionfactor gene (Oshox4and Oshox6) and IPT (isopentenyltransferases) gene have a potential value ofgenetic improvement of rice drought tolerance, at the end for the three gene we did a more in depthstudy. main results are as below:1.35S::Oshox4PCAMBIA vector were constructed, through Agro bacteriumtumefactions-mediated the gene transfer to drought stress sensitive variety IR64, and54positive plantswere obtained. by southern blotting analyze from54transgenic events19individual events were singlecopy,were confirm individual plants was positive, for the19individual single copy events and wild typeplant carried out phenotype observation and analyze.2. Selected two transgenic single copy events exposed to progressive soil drying at flowering stageand measured some drought index, to confirm Oshox4gene function in the drought stress condition. Andsame time we carried out phenotype observation for6single copy T2generation plants, found thatcompare to wild type all the transgenic events number of tiller and panicle were increased, but plantheight and internodes were shorter than wild type, flowering date is delayed and grains were unfilled.3. Using TAIL-PCR method found gene insert position in the chromosome and depend on geneinsert position design specific primer for the each line determine homozygous plants form T1generation.through phenotype observation found that in the same line between homozygous andheterozygous also have a obvious difference.4. Constructed PCAMBIA1300Oslea::Oshox6drought induce able vector, through agrobacterium tumefactions-mediated transform to drought sensitive rice variety IR64,obtained86hygromycin resistance plants. Using PCR and Southern blot analyze confirm gene integration and copynumber. The three single copy T2generation plants were exposing drought stressed in the PVC tube, andobserved that three transgenic plants root length and root diameter were obviously higher than wildtype.In the field drought stress condition,transgenic plant's drought tolerance is better than wild type.5. Oshox6gene natural promoter was cloned and ligated with Gus gene,made a Oshox6promoter::Gus construct, Through Agro bacterium tumefactions-mediated transform to rice variety,using PCR confirm gene integration and processing Gus staining to the transgenic plant root, shoot leafand anther, we found that Oshox6promoter was specific expression in root endodermis. 6. SARK::IPT PCAMBIA1380drought induce able vector were constructed, through Agrobacterium tumefacients-mediated transform to rice variety IR64,obtained256hygromycin resistanceplant,by the PCR and Southern blot analyze to confirm gene integration and copy number.depend onSouthern blot analyze selected10single copy plants were selected for the phenotype observation. Andexposed to drought stress for the two transgenic plants, the result showed that in drought stress conditionIPT transgenic plants chlorophyll content and cytokine content were obviously higher than wild type.