利用VIGS技术研究<i>NtbHLH93</i>基因在烟草甾醇代谢中的功能

武明珠; 刘瑞霞; 王中; 郑庆霞; 张剑锋; 李泽锋; 谢小东; 魏攀; 李锋; 罗朝鹏; 曹培健; 杨军

doi:10.16135/j.issn1002-0861.2018.0400

利用VIGS技术研究NtbHLH93基因在烟草甾醇代谢中的功能

Study on function of NtbHLH93 in sterol metabolism of tobacco by VIGS

摘要

摘要: 为进一步研究烟草NtbHLH93基因的功能，利用病毒诱导基因沉默（Virus induced gene silence，VIGS）技术降低烟草中NtbHLH93基因的表达。通过测定pTRV2-NtbHLH93烟株中的甾醇含量，发现NtbHLH93基因表达下调后，总甾醇、豆甾醇、胆固醇和β-谷甾醇的含量显著降低。转录组测序发现NtbHLH93基因下调后，共有259个基因差异表达。对这些差异表达基因进行KEGG表达通路分析，发现质体萜类代谢2-C-甲基-D-赤藓糖醇-4-磷酸（2-C-Methyl-D-Erythritol-4-Phosphate，MEP）途径中有两个基因（Niben101Scf00063g13014.1和Niben101Scf06249g03002.1）上调表达，这两个基因与MEP途径中的牻牛儿基牻牛儿基还原酶（Geranylgeranyl diphosphate synthases，GGPPS）基因序列高度相似，推测当甾醇合成代谢通路受阻后导致Niben101Scf00063g13014.1和Niben101Scf06249g03002.1基因上调表达。

Abstract: To study the function of NtbHLH93, virus-induced gene silencing (VIGS) was used to downregulate NtbHLH93 expression in tobacco. The results showed that when NtbHLH93 expression was downregulated, the contents of total sterol, stigmasterol, cholesterol and β-sitosterol significantly decreased in pTRV2-NtbHLH93 tobacco seedlings. Transcriptome sequencing revealed that there were 259 differentially expressed genes in NtbHLH93 expression downregulated tobacco seedlings. KEGG pathway analysis indicated that two genes (Niben101Scf00063g13014.1 and Niben101Scf06249g03002.1) were upregulated in MEP (2-C-methyl-D-erythritol-4-phosphate) pathway, a terpenoid metabolism pathway in plastids. The sequences of the two genes were highly similar to geranylgeranyl diphosphate synthases (GGPPS) gene in MEP pathway, which suggested that the upregulation of Niben101Scf00063g13014.1 and Niben101Scf06249g03002.1 might relate to the inhibition of sterol synthesis.

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