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作物学报 ›› 2016, Vol. 42 ›› Issue (01): 1-10.doi: 10.3724/SP.J.1006.2016.00001

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

GmMYB042基因对类黄酮生物合成的调控作用

杜海1，冉凤1，马珊珊1，柯蕴倬1，孙丽萍1，李加纳1,*，唐益雄2,*   

  1. 1西南大学农学与生物科技学院，重庆400715；2中国农业科学院生物技术研究所，北京100081
  • 收稿日期:2015-06-08 修回日期:2015-09-06 出版日期:2016-01-12 网络出版日期:2015-10-08
  • 通讯作者: 李加纳, E-mail: ljn1950@swu.edu.cn, Tel: 13509496702; 唐益雄, E-mail: yixiongtang@sohu.com, Tel: 010-62121506.
  • 基金资助:

    本研究由国家自然科学基金项目(31471528)，国家博士后基金面上项目(2014M552297)和中央高校基本科研业务费专项资金(SWU113104, XDJK2014B035)资助。

Regulating Effects of GmMYB042 Gene on Flavonoid Biosynthesis

DU Hai1, RAN Feng1,MA Shan-Shan1,KE Yun-Zhuo1,SUN Li-Ping1,LI Jia-Na1,*,TANG Yi-Xiong2,*   

  1. 1 College of Agronomy and Biotechnology, Southwest University, Chongqing 400716, China; 2 Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2015-06-08 Revised:2015-09-06 Published:2016-01-12 Published online:2015-10-08
  • Supported by:

    This research was supported by National Natural Science Foundation of China (31471528), Postdoctoral Science Foundation of China (2014M552297) and Fundamental Research Funds for the Central Universities of China (SWU113104, XDJK2014B035).

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摘要：

MYB类转录因子是植物中最大的转录因子基因家族之一，广泛参与植物生长发育全过程，对植物次生代谢等具有重要的调控作用。本研究对大豆GmMYB042基因的表达特性和功能进行了系统的研究，针对该基因C端的保守氨基酸基序(PDLNLELTIS)和锌指结构进行了一系列的序列删除突变，并将各缺失突变体在烟草中进行了过表达，以验证目的基因及其特殊基序的功能。表达特性分析结果表明GmMYB042基因在大豆的根瘤、根、茎、叶、花、荚果皮和种子中均有表达，且在茎、种子和花中的表达量相对较高；GmMYB042基因在大豆中的表达受PEG、高盐、低温和UV-B辐射的诱导。过表达分析结果表明，GmMYB042基因的过表达使转基因烟草类黄酮代谢途径部分关键酶基因(如PAL、CHS和FLS)的表达量明显上升，转基因烟草总黄酮的含量明显高于对照；各缺失突变体的转基因烟草类黄酮代谢途径相应酶基因的表达量发生相应的变化，进一步证明目的基因对类黄酮生物合成的调控作用；各缺失突变体的转基因烟草的叶缘有明显的皱褶，说明目的基因可能还参与调控叶的形态建成。

关键词: MYB转录因子, 类黄酮, 缺失突变, 功能研究

Abstract:

MYB transcription factor is one of the largest transcription factor gene families in land plants, and is involved in a myriad of regulatory processes, such as secondary metabolism. In the present study, the expression profiles and function of GmMYB042 gene were systematically studied. In order to investigate the roles of the conserved amino acid motif PDLNLELTIS and a predicted zinc finger region at its C-terminal, a series of sequence deletions of these two regions were made by PCR method. Subsequently, the corresponding over-expression constructs of GmMYB042 gene and its mutants were made and transformed into tobacco NC89 with Agrobacterium LBA4404, respectively. Expression analyses revealed that GmMYB042 gene was expressed in nodule, root, stem, leaf, flower, pod and seed of soybean, and with a relative higher expression level in stem, flower and seed; its expression could be induced by PEG, high salt, low temperature and UV-B radiation stresses. Over-expression analyses showed that the expressions of some enzyme genes in flavonoid biosynthesis pathway (including PAL, CHS, CHI, and FLS) were obviously increased in GmMYB042 transgenic lines, resulting in an increased content of the flavonoid compounds. Accordingly, the transcription levels of the corresponding enzyme genes involved in flavonoid biosynthesis pathway were decreased in the transgenic lines of GmMYB042 mutants, further supporting the conclusion of regulating role of GmMYB042 gene in tobacco flavonoid biosynthesis pathway.

Key words: MYB transcription factor, Flavonoid, Deletion mutation, Functional research

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