The Construction Of Full-length Cdna Library Of Erianthus Arundinaceus Under Drought Stress And Elementary Analysis Of Hir Gene

Sugarcane is the most important sugar crop in China even in the world.At the recent decades the sugarcane planting area had to moved to relatively arid field even drought sloping field from fertile field.It makes sugarcane breeding focus on the research of new breeds having outstanding drought resistance and high rooting ability.Erianthus arundinaceus,the related genara of Saccharum officinarum,has good applicability,stress resistance,vigour and rationing ability.And therefore it is important gene resource for sugarcane breeding.We constructed the full-length cDNA library of the E.arundinaceus under the stress of drought.And then from among these library,we picked up some clones random to sequence the DNA.The hir gene of E.arundinaceus was gotten.It had the intact 5' and 3' ends,and has a cds of 855bp.After the analysis of bioinformatics,we presumed that it is related with the drougnt resistance of the plants.So we designed the primers of the hir gene for fluorescent-quantity PCR,and we found that the expression of hir was toned up along with the degree of drought stressed on the plants.The maximum of the expression could be 48.12 times on the normal level when treated for 72hr with 40% PEG4000.Thus,we constructed a new vector,named pRD-HIR,incluing the cds of hir gene,driven by drought-responsive promoter,Prd29A,based on the ready-made vector, pCRBI.And then transformed the Arabidopsis thaliana,screened the positive plant with the PPT.Through PCR test,we confirmed the CDS of hir had been cloned into the genome of Arabidopsis thaliana.For more confirment of hir gene related to drought resistance,we designed the primers to construct a new RNAi vector,based on the ready-made vector, pTCK-303,named pTCK-HIR.