甘蓝型油菜BnaA05.nsLTP2基因克隆及表达模式分析OA

Plant non-specific lipid transfer proteins(nsLTPs)are a type of small molecule soluble proteins that can transfer lipids between membranes and involve in multiple biotic and abiotic stress response.In our previous study,a non-specific lipid transfer protein,BnaA05.nsLTP2,that interact with key glycerol-3-phosphate acyltransferase,was identified by yeast two hybrid assay.In this study,the BnaA05.nsLTP2 gene was cloned from'Zhongshuang 11'variety of Brassica napus,and the length of coding sequence was 294 bp,encoding 97 amino acids,with a conserved domain of lipid transfer protein and 8 highly conserved cysteine residues.Phylogenetic analysis revealed that the sequence similarity between BnaA05.nsLTP2 and its homologous protein sequence in Brassica rapa reached 95.88%.In addition,the promoter region of BnaA05.nsLTP2 contained several cis-acting elements,including LTR(low-temperature response),ABRE(abscisic acid response),TCA-element(salicylic acid response),and so on.The pCAMBIA1305.1-35S-BnaA05.nsLTP2-GFP fusion expression vector was constructed,and the results of subcellular localization indicated that the protein was localized to cell membrane.The tissue-specific expression pattern analysis showed that the relative expression of the BnaA05.nsLTP2 gene was significantly higher in roots and stems than other tissues(P＜0.05).In addition,the BnaA05.nsLTP2 gene was induced by low temperature stress,osmotic and salt stress,suggesting that this gene might be involved in regulating the abiotic stress response mechanism of B.napus.This study provides theoretical basis for elucidating the biological function of BnaA05.nsLTP2 in B.napus.