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辣椒素主要在果实胎座组织中合成,为了进行辣椒素生物合成的分子遗传学研究,采用SSH法,构建一个特辣型品种Habanero胎座cDNA文库。通过dotblot和核苷酸序列分析,从400个消减克隆中选择了39条cDNA克隆,将其与数据库中DNA和蛋白质进行了比对,估计这些克隆的序列信息。结果表明cDNA克隆可分为4类:I类cDNAs,与编码包括酰基转化酶和脂肪酸乙醇氧化酶在内的代谢酶基因同源;Ⅱ类为推定细胞壁蛋白质;Ⅲ类为生物和非生物胁迫蛋白质;Ⅳ类为非同源cDNAs。Northern blot分析证明这些克隆在辣椒中差异表达。结果说明所有的cDNA克隆在辣椒中差异表达,Ⅰ类和Ⅳ类的cDNA克隆在辣椒胎座中差异表达或优先表达。
Capsaicin is mainly synthesized in the fruit placenta tissue. In order to study the molecular genetics of capsaicinoid biosynthesis, an extra-hot Habanero placenta cDNA library was constructed by SSH method. By dot blot and nucleotide sequence analysis, 39 cDNA clones were selected from 400 subtractive clones and compared with the DNA and protein in the database to estimate the sequence information of these clones. The results show that the cDNA clones can be divided into four classes: Class I cDNAs, which are homologous to genes encoding metabolic enzymes including acylase and fatty alcohol oxidase; class II is putative cell wall proteins; and class III are biotic and abiotic stress proteins ; Ⅳ is non-homologous cDNAs. Northern blot analysis demonstrated that these clones were differentially expressed in pepper. The results showed that all cDNA clones were differentially expressed in pepper, and the cDNA clones of class I and IV were differentially expressed or preferentially expressed in pepper placenta.