Genome-wide analysis, molecular cloning and expression profiling reveal tissue-specifically expressed, feedback-regulated, stress-responsive and alternatively spliced novel genes involved in gibberellin metabolism in Salvia miltiorrhiza.
Genome-wide analysis, molecular cloning and expression profiling reveal tissue-specifically expressed, feedback-regulated, stress-responsive and alternatively spliced novel genes involved in gibberellin metabolism in Salvia miltiorrhiza.
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DOI：
10.1186/s12864-015-2315-5
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发表时间：
2015-12-21
期刊：
影响因子：
4.4
通讯作者：
Lu S
中科院分区：
文献类型：
--
作者：
Du Q;Li C;Li D;Lu S
赤霉素（GA）是一种典型的植物激素，在植物生长发育中起着重要的作用。丹参（Salvia miltiorrhiza Bunge）是一种重要的中药材，也是一种新兴的模式药用植物，其主要的亲脂性生物活性成分二萜类丹参酮与其重要的中间体二磷酸前体GGPP是相同的。对GA代谢及其调控的分析有助于阐明GA的生物学功能以及GA代谢与丹参酮生物合成之间的相互影响。丹参。然而，参与对映贝壳杉烯转化为GA的基因尚未得到系统的研究。通过全基因组预测和分子克隆，首次系统鉴定了22个赤霉素代谢途径候选基因。它包括一个SmKO、两个SmKA 0、六个SmGA 20 ox、两个SmGA 3 ox和十一个SmGA 2 ox，其中二十个基因是新的。推导的蛋白质序列具有保守性和差异性。赤霉素代谢途径基因具有组织特异性表达模式，对外源GA 3处理的响应存在差异，表明赤霉素代谢在不同组织类型中存在差异调节。丹参。SmKAO 1、SmKAO 2、SmGA 2 ox 2和SmGA 2 ox 4-SmGA 2 ox 7基因表达显著上调，SmGA 20 ox 2、SmGA 3 ox 1、SmGA 2 ox 1、SmGA 2 ox 8、SmGA 2 ox 10和SmGA 2 ox 11基因表达显著下调;而其他基因的表达在不同组织类型和不同时间点的GA 3处理中存在差异，表明反馈调控的复杂性。丹参酮生物合成相关基因SmCPS 1和SmKSL 1在GA 3处理后表达上调。在22个已鉴定的基因中，有9个对酵母抽提物和Ag+处理有反应。丹参毛状根。此外，SmKO，SmGA 20 ox 3，SmGA 2 ox 3和SmGA 2 ox 11的组织特异性表达的剪接变体被鉴定，其中SmKOv 1，SmGA 20 ox 3v和SmGA 2 ox 11 v1是GA 3-响应的，这表明选择性剪接在调节GA代谢中的重要性。研究结果表明，这些新基因具有组织特异性表达、反馈调节、胁迫响应和选择性剪接等特性，揭示了赤霉素代谢的多层次调控以及赤霉素代谢与丹参酮生物合成之间的相互作用。丹参。本文的在线版本（doi：10.1186/s12864-015-2315-5）包含补充材料，可供授权用户使用。
Gibberellin (GA), a classical phytohormone, plays significant roles in plant growth and development. It shares the important intermediate diphosphate precursor, GGPP, with the main lipophilic bioactive components, diterpenoid tanshinones in Salvia miltiorrhiza Bunge, one of the most important Traditional Chinese Medicine materials and an emerging model medicinal plant. Analysis of GA metabolism and regulation may help to demonstrate the biological functions of GAs and the crosstalk between GA metabolism and tanshinone biosynthesis in S. miltiorrhiza. However, genes involved in the conversion of ent-kaurene to GAs have not been systematically studied. Through genome-wide prediction and molecular cloning, twenty two candidate gibberellin metabolism pathway genes were systematically identified for the first time. It includes a SmKO, two SmKAOs, six SmGA20oxs, two SmGA3oxs and eleven SmGA2oxs, of which twenty genes are novel. The deduced proteins showed sequence conservation and divergence. Gibberellin metabolism pathway genes exhibited tissue-specific expression patterns and responded differentially to exogenous GA3 treatment, indicating differential regulation of gibberellin metabolism in different tissue types in S. miltiorrhiza. SmKAO1, SmKAO2, SmGA2ox2, and SmGA2ox4–SmGA2ox7 were significantly up-regulated; SmGA20ox2, SmGA3ox1, SmGA2ox1, SmGA2ox8, SmGA2ox10 and SmGA2ox11 were significantly down-regulated; while the responses of many other genes varied among different tissue-types and time-points of GA3 treatment, suggesting the complexity of feedback regulation. Tanshinone biosynthesis-related genes, such as SmCPS1 and SmKSL1, were up-regulated in response to GA3 treatment. Among the 22 identified genes, nine responded to yeast extract and Ag+-treatment in S. miltiorrhiza hairy roots. Moreover, tissue-specifically expressed splice variants were identified for SmKO, SmGA20ox3, SmGA2ox3 and SmGA2ox11, of which SmKOv1, SmGA20ox3v and SmGA2ox11v1 were GA3-responsive, suggesting the importance of alternative splicing in regulating GA metabolism. The results show tissue-specifically expressed, feedback-regulated, stress-responsive and alternatively spliced novel genes and reveal multiple layer regulation of GA metabolism and crosstalk between gibberellin metabolism and tanshinone biosynthesis in S. miltiorrhiza. The online version of this article (doi:10.1186/s12864-015-2315-5) contains supplementary material, which is available to authorized users.