Construction of yeast cDNA library of Camellia oleifera induced by ethylene and screening of CoFAD7 upstream regulators
Exogenous ethylene could promote the accumulation of α-linolenic acid(ALA)in Camellia oleif-era seeds,FAD7 is a key gene affecting ALA synthesis.In order to elucidate the molecular regulatory mech-anism of CoFAD7 expression in C.oleifera,this study cloned the promoter of CoFAD7 and constructed yeast one hybrid cDNA library of C.oleifera seeds induced by ethylene.The upstream regulatory factors that bind to the CoFAD7 promoter were screened through yeast one hybrid technology.The results indicate that the 1 395 bp CoFAD7 promoter sequence contains multiple cis-elements and transcription factor bind-ing elements related to hormone regulation and environmental stress.The constructed cDNA library had a capacity of 9.5×106 CFU·mL-1 and an average insertion fragment length of approximately 1 000 bp.Four transcription factors AP2-3,NFYC-1,F-box and ZFP1 were obtained through yeast one hybrid screening,which can bind to the CoFAD7 promoter.The above results lay the foundation for further exploring the mo-lecular mechanism of CoFAD7 gene responsing to ethylene on regulation of ALA content.
Camellia oleiferathe promoter of CoFAD7yeast one hybridcDNA librarytranscription factor
NETL
NSTL
万方数据
