小菜蛾MAP4K4基因上游5′-侧翼序列克隆与结构分析  

Cloning and structure analysis of the 5′-flanking sequence of MAP4K4 gene from the diamondback moth,Plutella xylostella

作　　者：郭乐 郭兆将[2] 康师[2] 孙丹[2] 周君雷 龚莉君[1,2] 张友军 GUO Le;GUO Zhaojiang;KANG Shi;SUN Dan;ZHOU Junlei;GONG Lijun;ZHANG Youjun(College of Plant Protection,Hunan Agricultural University,Changsha　410128,China;Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing　100081,China)

机构地区：[1]湖南农业大学植物保护学院,长沙410128 [2]中国农业科学院蔬菜花卉研究所,北京100081

出　　处：《植物保护》2018年第2期38-46,74,共10页Plant Protection

基　　金：国家重点研发计划项目(2017YFD0201204);国家自然科学基金(31630059;31701813);蔬菜有害生物控制与优质栽培北京市重点实验室和中国农业科学院科技创新工程(AAS-ASTIP-IVFCAAS)

摘　　要：本实验室前期研究表明,小菜蛾通过有丝分裂原激活的蛋白激酶(mitogen-activated protein kinase,MAPK)信号途径上游转录激活的关键基因MAP4K4反式调控多个中肠受体基因的表达,从而介导其对苏云金芽胞杆菌Bacillus thuringiensis(Bt)产生的Cry1Ac杀虫蛋白的高抗性。为进一步明确MAP4K4基因转录激活而过量表达的顺式调控机制,本文利用小菜蛾基因组数据库中MAP4K4基因序列信息,首先克隆了Bt Cry1Ac敏感小菜蛾种群MAP4K4基因上游5′-侧翼序列,得到了两种不同形式的5′-侧翼序列:MAP4K4-1和MAP4K4-2。随后,预测分析了其中的潜在功能顺式作用元件,同时发现其中核苷酸的转换会导致顺式作用元件的改变。本研究初步揭示了小菜蛾MAP4K4基因的5′-侧翼序列的遗传多样性,为后续明确MAP4K4的顺式调控机制奠定了基础。Our previous studies indicated that the crucial gene MAP4K4,a constitutive transcriptionally-activated upstream gene of MAPK signaling pathway,could trans-regulate the expression of multiple midgut receptor genes thereby mediating the high-level resistance to Bt Cry1Ac toxin in Plutella xylostella(L.).In this study,to further clarify the cis-regulatory mechanism of transcriptional activation and overexpression of MAP4K4 gene,we utilized the genome database of P.xylostella and cloned the 5′-flanking sequence of MAP4K4 gene from Bt Cry1Ac-susceptible P.xylostella for the first time,and two isoforms including MAP4K4-1 and MAP4K4-2 were detected.Subsequently,the potential functional cis-acting elements in both MAP4K4-1 and MAP4K4-2 isoforms were predicted,and the results showed that the transformation between nucleotides could result in alterations of intrinsic cis-elements.This study revealed the genetic diversity of the 5′-flanking sequence of MAP4K4 gene in P.xylostella and lays a foundation for further exploration of the cis-regulatory mechanism of MAP4K4 gene in P.xylostella.

关 键 词：小菜蛾 MAPK信号途径 MAP4K4 5′-侧翼序列 顺式调控机制 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]