Molecular cloning and expression analysis of ChCAO gene in leaf art formation of Cymbidium hybrid
[Objective]Chlorophyllide a oxygenase(CAO)is a key enzyme in the formation of chlorophyll b.Cloning and expression analysis of CAO genes may be helpful in exploring their function in regulating leaf color variation of Cymbidium hybrid.[Methods]ChCAO gene was cloned from leaves of Cymbidium hybrid by RACE and RT-PCR.ChCAO was analyzed for structure characteristics,physicochemical prop-erties,sequence alignment,and phylogenetic relationship.The expression of ChCAO in different tissues and leaves of K21-3 were analyzed by qRT-PCR.Meanwhile,ChCAO expression was silenced by VIGS technology.[Results]The results showed that ChCAO coding region was 1 608 bp,encoding 535 amino acids.ChCAO was closely related to Cymbidium sinense,which belonged to genus Cymbidium.qRT-PCR assay showed that the expression of ChCAO was the highest in leaves and the lowest in roots.The expres-sion of ChCAO in K21 leaves was significantly higher than that of K21-3.The VIGS silencing vector of ChCAO gene was constructed and transferred to tobacco.The viral vector was successfully invaded the to-bacco,and the old leaves of tobacco were turned yellow,while chlorophyll content and expression of ChCAO gene were reduced.[Conclusion]The ChCAO gene was obtained and its function was preliminari-ly characterized,which are helpful for the understanding of the formation of leaf art in Cymbidium hybrid.
NETL
NSTL
万方数据
维普
