The Study On The Function Of Three Novel Phospholipase D Genes In Response To Low Temperature Stress In Transgenic Tobacco

The abitotic stresses serve as the important environmental factors that influence the normal growth of plants.Among these factors,the stress of low temperature is one of the most harmful factors.The low temperature can cause the damage in different degrees, even more death in plants. Plants develop a variety of sophisticated mechanisms,which including the effect of low temperature response genes and the signal pathway,to response the changing environment and protect themselves from environmental stress.In this study,we uesd Chorispora Bungeana which is a rare alpine subnival plant species as material. The technology of genetically modified and virus-induced gene silencing(VIGS) were used to verify the functionalities of CbPLD?, CbPLD?,CbPLD? gene from transgenic tobacco,and analysis the response mechanisms under low temperature stress.The main results are as follows:(1) Introduced CbPLD? into tobacco with the CaMV 35 S promoter. The transgenic tobacco plants were detected by PCR.The sequences results showed that the CbPLD? genes have been transferred into tobacco successfully.(2) Constructed a TRV-CbPLD?,TRV-CbPLD? and TRV-CbPLD? virul vectors and introduced into tobacco by Agrobacterium-mediated injection.The RT-PCR were used to detect the gene silencing rate of CbPLD?,CbPLD? and CbPLD? in transgenic tobacco.Then,we filtered the highest rate of gene silencing tobacco and named as TRV-CbPLD?1,TRV-CbPLD? 3 and TRV-CbPLD?2,respectively.(3) The expression levels of CbPLD?, CbPLD? and CbPLD? gene in transgenic tobacco under low temperature stress(4?) were studied.The result showed that,low temperature induced the expression of CbPLD?,CbPLD? and CbPLD?,and the expression of CbPLD? gene behind the CbPLD? and CbPLD? with the time prolong under low temperature stress.(4) The metabolism regulations of CbPLD transgenic tobaccos and TRV-CbPLDgene silence tobaccos in response to low temperature stress were analyzed. The results showed that both CbPLD transgenic tobaccos and TRV-CbPLD gene silence tobaccos involved in the regulation pathway of membrane stability, osmotic adjustment and the regulation of anti-oxidative system. CbPLD?, CbPLD? and CbPLD? gene can provide powerful genetic resources for improving low temperature resistance of plants.