Early Japonica Kongyu131 Barriers Chilling Tolerance Gene OsPGR5 Clone

Abiotic stress such as low temperatures, drought and salt is resulting in a decline incrop production, the quality and the main factors of limiting crop distribution, especiallythe low temperature stress can cause the production of crops, which is a commonproblem worldwide. As resistance molecular mechanism research of increasinglydeepening, the researchers found that some gene expression quantity has changedsignificantly under adversity stress, these genes are genetic transformation can improveplant resistance to inverse directly, thus more significantly improved crops resistanceeffect.Kongyu131is widely grown in the northeast of rice varieties, has characteristic ofhardy and high yield. Using Solexa sequencing chip to analysis the cold treatment ofRice Kongyu131booting in different times, to explore the molecular mechanismregulation of stress response to cold stress. From the expression profiling results, weselected out a gene-the proton gradient protein gene PGR5.It involved in photosynthesisand the expression quantity has changed significantly under adversity stress. Theexpression of OsPGR5gene was analyzed under cold stress at different times throughRealtime-PCR experiments. After gene cloning and genetic transformation of rice ofLonggeng11，finally transgenic plant was obtained. This research has great researchvalue for improving plant stress tolerance and also provides a theoretical basis and generesources for the application of cold resistance genetic engineering of crop. The mainresearch results are as follows:1. Booting stage of rice Kongyu131chilling digital expression profilingThe results showed that7180of differentially expressed genes had been discovered.There are225differentially expressed genes have been found in four times of treatment.By pathway analysis, when subjected to cold stress.Metabolic pathways associated withphotosynthesis, and glucose metabolism associated with regulation related, such asresistance to low temperature stress involved in major metabolic processes.2. OsPGR5gene expression analysisUsing semi-quantitative Real Time PCR method in processing to cold stress in ricematerial purpose gene expression quantity changes for testing, the results show that OsPGR5gene transient expression under cold stress, within24hours of cold stressresistance to cold rice Kongyu131OsPGR5gene expression continuously, Longjing11OsPGR5gene expression without regularity, the differences of two rice cold resistanceis likely as PGR5transcriptional regulation of gene expression level difference iscaused.3. The OsPGR5gene cloning and sequence analysisRice with cold treatment5h of Kongyu131cDNA as template, by PCRamplification, cloning OsPGR5genes related to cold stress, the gene cDNA sequence is760bp, including CDS area, a total of378bp, encoding125amino acids.4. The construction of plant expression vector and genetic transformationConnect OsPGR5gene to the plant expression vector pH7WG2by Gatewaycloning technology, by the method of agrobacterium mediated with callus, genetictransformation cold sensitivity rice of Longgeng11, fit for T0generation of transgenicrice molecular biological detection, conversion efficiency was8.4%, further determineOsPGR5gene is important in plant adversity stress resistance genes.