Study On Relationship Between The Expression Of Cold Related Protein Genes And Moisture State And Cold Resistance Of Winter Wheat

The freeze injury has become the main factor restrict the yield of winter wheat in Heilongjiang province as a result of the the cold weather in the winter and annual snowfall was imbalance.So,Screening out the cold resistance varieties will be significant to promote the winter wheat yield as well as its grown in high latitude.To reveal the water adaptation machine of winter wheat tillering node in over-wintering period and regulatory mechanism of genes related to cold resistance protein,six different cold resistant winter wheat varieties were used in the current study to detect the determination of the membrane stability of tillering nodes throughout the over-wintering period,moisture content and its composition and greening rate.Membership function was used to appraise the cold resistance of six winter wheat varieties.Then take the three strong cold resistance winter wheat varieties Dongmai 1,Dongnong 377,Dongnong 256 that obtained from the identification experiment as materials,Jimai 22(weak cold esistance)as control detecting the expression quantity of different genes about cold resistance protein.We filtered eight cold-related protein genes randomly from the digital gene expression profiling of wheat to determine the expression quantity of different cold-related protein genes in over-wintering period between winter wheat varieties using fluorescent quantitative PCR,in order to reveal the expression of cold-related protein genes of different winter wheat varieties in over-wintering period and provide basic reference for further studies such as gene cloning and breeding. Achieved the following:1.Under natural overwinter condition,the greening rate showed very significant negative correlation with conductivity,free water content and free water/bound water and Significant positive correlation with relative water content meanwhile kept a significant negative correlation with tissue water content.The relative water content and free water/bound water could better reflect the condition of cell membrane damage would be suitable traits to estimate the winter wheat cold resistance.2.The result of comprehensive evaluation of cold resistance of six winter wheat varieties in natural over-wintering period using membership function is that Dongmai 1,Dongnong 377 and Dongnong 256 was strong cold resistance varieties(levelⅠ),Dongnong 373 was cold resistance variety(level Ⅱ),Dongnong 608 act as a mid-resistance variety(level Ⅲ) and Jimai 22 was a weak resistance variety(level Ⅳ).3.Low temperature can induce the high expression of winter wheat disease resistance protein TSN1 gene,the negative accumulated temperature and the induction time TSN1 needed to express was different between varieties.The strong cold resistance varieties.shorter induction time and less negative accumulated temperature relatively,whereas TSN1 need longer induction time and more negative accumulated temperature to express for the weak cold resistance varieties.4.The over expression of the gene coding cold-acclimation protein WCOR518 and gene of cold temperature induced protein COR39 in strong cold resistance varieties could be induce by low temperature.However the the change of the gene expression quantity was not that significant in weak cold resistance varieties.5.Low temperature could induce the expression of dehydration responsive protein gene DBP and dehydrated protein gene DHN5 but the expression quantity of DBP was highest at mid frozen stage and DHN5 at deep frozen stage. In addition the expression quantity of DBP showed significant positive correlation with cold resistance.6.Low temperature could induce the expression of gene coding resistant protein NBS-LRR1 but the highest expression quantity occurred in different stages.The highest expression of DBP was at mid frozen stage and has significant negative correlation with cold resistance.In the over-wintering period,there were few law of the expression quantity of resistance protein NBL1 gene,further replication experiment still need do to explain the cold resistance of winter wheat controlled by NBL1.7.Gene of cold shock protein CS120 was sensitive to temperature,expression quantity of the gene was larger in strong cold resistance varieties than that in weak cold resistance varieties.What’s more the expression quantity of the gene coding CS120 was in proportion to the increase of negative accumulated temperature.