中国水仙NtMYB8的分离及功能研究

• 花匠小妙招
• 2024-09-16 20:02

摘要：

从中国水仙（Narcissus tazetta var. chinensis）‘金盏银台’中分离获得1个MYB转录因子基因,命名为NtMYB8,GenBank登录号:MF5222090。通过多重序列比对和进化树分析发现,NtMYB8属R2R3-MYB家族,与原花青素激活因子亲缘关系最近;NtMYB8在中国水仙副冠中表达量最高;将NtMYB8在普通烟草瞬时转化,注射的叶片出现微弱红色斑点;转35S-NtMYB8烟草花瓣颜色变化不明显,qPCR检测显示NtMYB8促进转基因烟草类黄酮代谢途径大部分结构基因表达;酵母双杂试验表明NtMYB8能与中国水仙NtbHLH1互作;双荧光素酶瞬时表达试验显示NtMYB8与NtbHLH1互作能够显著促进水仙类黄酮代谢途径关键基因NtFLS的启动子活性。研究结果表明,NtMYB8是中国水仙类黄酮代谢途径的激活因子,可能促进副冠中黄酮醇的合成。

关键词: 水仙, R2R3-MYB, 类黄酮代谢, 激活因子, 结构基因

Abstract:

A MYB transcription factor gene,named NtMYB8,was obtained from Chinese narcissus （Narcissus tazetta var. chinensis）. GenBank accession number:MF5222090. Multiple protein sequence alignment and phylogenetic tree analysis showed that NtMYB8 contained R2 and R3 conserved domains,belonging to the R2R3-MYB family,and was closely related to proanthocyanidin activators. The expression level of NtMYB8 in the corona was the highest. In the transient transformation of common tobacco,the leaves injected with NtMYB8 had slight red spots. However,the petal color of NtMYB8- transgenic tobacco did not change significantly,and further analysis by qPCR showed that NtMYB8 promoted the expression levels of most tobacco structural genes in the flavonoid metabolic pathway. Yeast two-hybrid assay showed that NtMYB8 can interact with NtbHLH1,the bHLH protein of Chinese narcissus and significantly promote the Chinese narcissus FLS promoter activity by dual luciferase transient expression test. These results suggest that NtMYB8 is an activator of the flavonoid metabolism pathway in Chinese narcissus and may promoter the flavonol biosynthesis in corona.

Key words: Narcissus tazetta var. chinensis, R2R3-MYB, flavonoid metabolism, activator, structural gene

中图分类号: 

S682.21

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