Cloning And Functional Verification Of GhKLCR1 Gene,Kinesin Light Chain-Related Gene,from Gossypium Hirsutum

With the change of unstable environments and increasing climatic fluctuations,drought stress has become one of the mostserious factors that affect greatly on globla crop productions,especially leading to the reduced production of cotton fiber.Though there are few reports on drought resistance of plants,genes,metabolomic pathways and their functional roles in the mechenisms for drought responses remain unclear.In present study,the gene GhKLCR1(kinesin light chain-related protein-1),which is the homologous gene of At KCLR1,was cloned from Gossypium hirsutum.and the related functional verification experiments were perfomed to explore role of GhKLCR1.It encodes a kinesin light chain-related protein,which is a poly-protein,however,the KLCR1 gene in cotton has not been reported yet.This study analyzed the expression pattern of GhKLCR1 gene and its function.The GhKLCR1 gene was cloned for the first time,and then the plant expression vector,the promoter-driven GUS(glucuronidase)reporter vector,and a tag vector of green fluorescent protein(GFP)were constructed.The constructed vectors were introduced into wild-type A.thaliana and positive transgenic lines were obtained and its function was explored through experiments.With the q RT-PCR experiments,GhKLCR1 was expressed higher in root than that of other tissues in upland cotton.Further abiotic and hormonal stress treatment showed that GhKLCR1 expression level was significantly up-regulated after PEG treatment,and it was found that GhKLCR1 was responsed with the treatment of salicylic acid(SA),brassinosteroid(BL)and auxin(IAA).In addition,GUS staining of the GhKLCR1 promoter-driven GUS transgenic Arabidopsis plants revealed that the degree of staining increased with increasing mannitol concentration.These results above suggested that GhKLCR1 is involved in stress response.After overexpressing GhKLCR1,the root lengths of 35S::GhKLCR1 lines are significantly shorter than that of Col-0.And the germination of 35S::GhKLCR1 lines seeds was strongly inhibited,indicating that overexpression of GhKLCR1 is more sensitive to drought.subcellular localization experiments showed that GhKLCR1 may be associated with cell membrane.These findings provide a better understanding of the structural,physiological,and functional mechanisms of the GhKLCR1 protein,and lay the foundation for the analysis and molecular improvement of the stress resistance mechanism in cotton.