生物技术通报 ›› 2020, Vol. 36 ›› Issue (12): 1-11.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0563

• 研究报告 •    下一篇

玉米萜类合成酶基因tps2的功能及其启动子功能区段鉴定

王丹1,2(), 李圣彦2(), 刘进平1(), 郎志宏2   

  1. 1.海南大学热带作物学院,海口 570100
    2.中国农业科学院生物技术研究所,北京 100081
  • 收稿日期:2020-05-12 出版日期:2020-12-26 发布日期:2020-12-22
  • 作者简介:王丹,女,硕士研究生,研究方向:农业生物技术;E-mail:748736649@qq.com
  • 基金资助:
    国家自然科学基金项目(31601702);转基因新品种培育重大专项(2019ZX08010-004)

Study on the Function of Terpene Synthase Gene tps2 and Its Promoter Functional Segment in Zea mays

WANG Dan1,2(), LI Sheng-yan2(), LIU Jin-ping1(), LANG Zhi-hong2   

  1. 1. College of Tropical Crops,Hainan University,Haikou 570100
    2. Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081
  • Received:2020-05-12 Published:2020-12-26 Online:2020-12-22

摘要:

虫害是造成玉米减产和品质下降的主要原因,玉米萜类合成酶基因tps2(terpene synthase 2)受虫害诱导表达,催化产生多种萜类化合物。qPCR检测不同激素和黏虫处理条件下tps2的表达;过表达植株(TPS2OE)进行玉米螟行为选择实验;原生质体瞬时表达检测tps2启动子的活性。结果显示植物激素(JA、SA、ET)和黏虫诱导tps2高表达;TPS2OE中tps2的表达量及目的萜类含量显著高于对照B73;玉米螟行为选择实验结果表明,尽管实验组和对照组叶片上的玉米螟卵的个数无显著差异,但TPS2OE叶片上的幼虫数较少,危害面积也更小;启动子截短分析发现-183至-129为tps2启动子必需的功能区段,并且-227至-183区段可能存在负调控元件。综上,tps2受植物激素和虫害诱导表达,TPS2OE植株对玉米螟幼虫有一定的抑制作用,tps2启动子必需的功能区段为-183至-129。

关键词: 玉米, tps2, 萜类化合物, 植物激素, 亚洲玉米螟, 启动子

Abstract:

Insect pest is the main reason causing the yield and quality of corn reduced. The tps2(terpene synthase 2)gene is inducible by herbivore and TPS2 catalyzes the production of many terpenoids. To analyze the function of tps2 and its expression regulation mechanism,qPCR was used to detect the expression of tps2 under different hormones and armyworm treatments. The transgenic maize TPS2OE was obtained by Agrobacterium-mediated transformation using a tps2 overexpression vector and it was used to study the selective behavior of Asian corn borer. The functional segment of tps2 promoter was tested by transient expression system of maize protoplast. The results showed that plant hormones(JA,SA,and ET)and armyworm induced high expression of tps2;and the expression of tps2 in TPS2OE and the target terpenoids were significantly higher than that in control B73. The number of corn borer eggs on the leaves of TPS2OE and B73 was insignificant,while the larvae on TPS2OE were fewer,and the damage areas were also smaller. The promoter truncation analysis found that the required functional element of the tps2 promoter was between -183 to -129,and there were repressor binding sites between -227 to -183. In summary,tps2 expression is induced by plant hormones and armyworm,and TPS2OE plants have a certain inhibitory effect on the larvae of Asian corn borer. The required functional element of tps2 promoter is between -183 to -129.

Key words: maize, tps2, terpenoids, plant hormones, Asian corn borer, promoter