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桂花OfMYB12基因参与类黄酮合成的功能分析OA北大核心CSTPCD

Functional Analysis of OfMYB12 Gene Involved in Flavonoid Synthesis in Osmanthus fragrans

中文摘要英文摘要

MYB转录因子广泛参与植物生长发育,对植物次生代谢具有重要调控作用.本课题组前期通过转录组测序(RNA-Seq)技术筛选出与桂花(Osmanthus fragrans)类黄酮合成相关的基因OfMYB12.本研究进一步从桂花'堰虹桂'(O.fragrans'Yanhonggui')的花芽克隆出OfMYB12 基因(GenBank No.WMD01176),该基因ORF全长为1 026 bp,编码341个氨基酸;亚细胞定位结果表明该基因定位于细胞核中.多序列比对和系统发育树分析表明,OfMYB12基因属于典型的R2R3转录因子,且与拟南芥(Arabidopsis thaliana)中的原花青素AtMYB56激活因子亲缘关系最近.组织特异性结果分析表明,OfMYB12基因在桂花不同组织中均表达,并且在花瓣中表达量最高;在白花矮牵牛'米歇尔'(Petunia hybrida'Mitchell')中异源过表达OfMYB12,结果发现,转基因花瓣中的总黄酮含量显著高于对照组(P<0.05),并且在类黄酮合成通路上的相关基因(PhCHS(Petunia hybrida chalcone synthase),PhF3H(P.hybrida flavanone 3-hydroxylase),PhFLS(P.hybrida flavonol synthase))的表达量显著升高.基于以上结果,推测OfMYB12基因可能参与类黄酮次生代谢生物合成的调控.本研究结果为桂花类黄酮合成分子机制探索及桂花品质改良提供了理论依据.

MYB transcription factor is widely involved in plant growth and development,and plays an important role in regulating plant secondary metabolism.The OfMYB12 gene,which was related to flavonoid synthesis and screened by RNA-Seq technology in previous study of our research group,was further cloned from the flower buds of Osmanthus fragrans'Yanhonggui'.The ORF length of OfMYB12 gene(GenBank No.WMD01176)was 1 026 bp and encoded 341 amino acids.The subcellular localization results indicated that this gene was located in the nucleus.Multiple sequence alignment and phylogenetic tree indicated that the OfMYB12 gene belonged to a typical R2R3 transcription factor and had the closest relationship with anthocyanin activator AtMYB56 in Arabidopsis thaliana.The analysis of tissue-specific results showed that OfMYB12 gene was expressed in different tissues of O.fragrans and had the highest expression level in petals.Heterologous overexpression of OfMYB12 in Petunia hybrida'Mitchell'resulted in a significantly higher total flavonoid content in the transgenic petals compared to the control group(P<0.05),and the expression levels of related genes(PhCHS(P.hybrida chalcone synthase),PhF3H(P.hybrida flavanone 3-hydroxylase),PhFLS(P.hybrida flavonol synthase))in the flavonoid synthesis pathway in transgenic petals were significantly increased.Based on the above results,it is speculated that the OfMYB12 gene might be involved in the regulation of flavonoid secondary metabolism biosynthesis.The results of this study provides a theoretical basis for exploring the molecular mechanism of flavonoid synthesis in Osmanthus and improving the quality of Osmanthus.

张慧莲;王艺光;钟诗蔚;邓金萍;方遒;赵宏波

浙江农林大学风景园林与建筑学院/浙江园林植物种质创新与利用重点实验室/国家林业和草原局南方园林植物种质创新与利用重点实验室,杭州 311300

园艺学与植物营养学

桂花花瓣OfMYB12基因总黄酮

Osmanthus fragrans petalsOfMYB12geneTotal flavonoids

《农业生物技术学报》 2024 (005)

1037-1048 / 12

国家自然科学基金面上项目(32072615);浙江省农业新品种选育重大科技专项(2021C02071)

10.3969/j.issn.1674-7968.2024.05.005

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