马铃薯萜类合酶基因StHcS的功能鉴定
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四川省“千人计划”青年人才项目


Functional Characterization of Terpene Synthase Gene StHcS in Solanum tuberosum
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    摘要:

    为了研究马铃薯萜类代谢物的生物合成机制,从马铃薯基因组数据中筛选到一个萜类合酶基因。通过RTPCR方法,从致病疫霉侵染后的马铃薯品种‘费乌瑞它’中成功克隆到该基因,命名为StHcS,并对其进行生物信息学分析、生化功能鉴定及表达模式分析。结果表明:(1)序列分析显示StHcS编码区序列长1 497 bp,编码498 个氨基酸,分子质量为74.78 kD。(2)StHcS基因编码的蛋白序列含有DDXXD催化功能域,与短柱茶(Camellia maliflora)中的四甲基环癸二烯甲醇合酶相似度最高。(3)蛋白体外催化实验和大肠杆菌代谢工程分析表明,StHcS可以催化生成倍半萜化合物四甲基环癸二烯甲醇(hedycaryol)。(4)基因表达分析显示,StHcS可以被致病疫霉侵染诱导表达反式法尼烯焦磷酸(FPP),且在侵染后72 h表达最高;GCMS分析显示,在受侵染的马铃薯块茎中检测到四甲基环癸二烯甲醇。StHcS生化功能的鉴定为倍半萜合酶的研究提供了多样性,也是首次在马铃薯中发现的四甲基环癸二烯甲醇合酶,为马铃薯萜类代谢途径解析提供了参考。

    Abstract:

    To elucidate the biosynthetic pathways of potato terpenoids, we identified a terpene synthase gene named as StHcS from potato genome data. StHcS was successfully cloned from potato variety ‘Feurita’ with infection of Phytophthora infestans by RTPCR. Bioinformatics analysis, biochemical characterization and gene expression pattern analysis were conducted for StHcS. The results are as follows: (1) the coding sequence length of StHcS is 1 497 bp and it eventually encodes 498 amino acids with a molecular mass of 74.78 kD; (2) the encoded StHcS contains the conserved DDXXD catalytic motif, and exhibits the highest similarity to the hedycaryol synthase in Camellia maliflora; (3) in vitro assay and E. coli metabolic engineering experiments reveal that StHcS can react with trans farnesene pyrophosphate (FPP) to produce the sesquiterpenehedycaryol; (4) RTPCR analysis indicates that StHcS can be induced by infection of P. infestans in potato tubers and the highest expression was observed at 72 h post inoculation. Hedycaryol was also successfully detected in the infected potato tubers by GCMS analysis. The identification of StHcS biochemical function provides diversity for the studies regarding sesquiterpene synthase, and it is also the first hedycaryol synthase found in potato, which provides a reference for the analysis of potato terpene biosynthesis.

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陈懿瑶,王丽平,谌琴琴,等.马铃薯萜类合酶基因StHcS的功能鉴定[J].西北植物学报,2020,40(7):1097-1104

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  • 在线发布日期: 2020-08-24
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