Q型烟粉虱UDP-糖基转移酶UGT354A1 基因克隆及其在噻虫嗪抗性中的作用
Cloning of the Bemisia tabaci Q biotype UDP-glycosyltransferase UGT354A1 gene and its role in thiamethoxam resistance
杜田华 危学高 殷 城 杨 静 黄明娇 桂连友 杨 鑫 张友军
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DOI:10.7679/j.issn.2095-1353.2022.028
作者单位:长江大学农学院昆虫研究所,荆州 434025;中国农业科学院蔬菜花卉研究所,北京100081
中文关键词:Q型烟粉虱;UDP-糖基转移酶(UGTs);UGT354A1基因;基因克隆;RNAi;噻虫嗪抗性
英文关键词:Bemisia tabaci; UDP-glycosyltransferases (UGTs); UGT354A1; gene cloning; RNA interference; thiamethoxam- resistance
中文摘要:
【目的】 UDP-糖基转移酶(UGTs)是昆虫主要的Ⅱ期解毒酶,可能参与昆虫抗药性的形成。本研究旨在探究Q型烟粉虱中UGT基因是否参与其对噻虫嗪的抗药性。【方法】 依据烟粉虱基因组数据库设计引物,克隆其UGT354A1基因的全长序列;采用qRT-PCR 技术检测UGT354A1基因在烟粉虱不同发育阶段、不同组织部位以及抗敏品系中的表达量;通过RNA干扰试验,验证UGT354A1基因在烟粉虱噻虫嗪抗性中的作用。【结果】 序列生物信息学分析发现,UGT354A1基因属于典型的昆虫UGTs,包括两个糖基供体(DBR1和DBR2)和一个保守特征性基序。qRT-PCR结果显示,UGT354A1基因在噻虫嗪抗性品系(THQR)中的表达量为敏感品系(THQS)的2.60倍,且噻虫嗪可显著诱导该基因的表达。在不同发育阶段和组织部位中,发现UGT354A1基因在烟粉虱若虫和成虫阶段及头部和胸部表达丰富。RNAi后,沉默UGT354A1基因能够显著增加噻虫嗪对Q型烟粉虱的毒杀作用。【结论】 UGT354A1基因在Q型烟粉虱中对噻虫嗪具有重要的解毒作用,可能参与了抗性的形成。
英文摘要:
[Objectives] To
investigate whether the Bemisia tabaci (Q-biotype)
UDP-glycosyltransferase (UGT) gene is involved in resistance to thiamethoxam. [Methods] PCR primers were designed according
to the genome database and the full-length sequence of the UGT354A1 gene
was cloned. Expression levels of the UGT354A1 gene in different
developmental stages and body parts, and in a thiamethoxam-resistant (THQR) and
susceptible strain (THQS) were detected by q-PCR. The function of UGT354A1 gene in the resistance of B. tabaci to thiamethoxam was verified by
RNAi. [Results] Sequence analysis
shows that the UGT354A1 gene has a
conserved UGTs domain that is typical of insect UGTs. Compared with the
susceptible strain, the UGT354A1 gene was overexpressed (2.60-fold) in
the thiamethoxam resistant strain. In addition, thiamethoxam could
significantly induce the expression of the UGT354A1 gene. The experiments
of the expression levels of the UGT354A1 gene in different developmental
stages and body parts indicate that it is most highly expressed in nymphs and
adults, and in the head and thorax. RNAi silencing of UGT354A1 gene
expression significantly reduced the resistance of B. tabaci to
thiamethoxam. [Conclusion] The UGT354A1 gene is involved in the formation of thiamethoxam resistance in B. tabaci. The
results improve the understanding of the role of UGTs in the evolution of
insecticide resistance, and provide a theoretical basis for the comprehensive
management of B. tabaci.