Tan Y-A, Zhao X-D, Sun Y, Hao D-J, Zhao J, Jiang Y-P, Bai L-X, Xiao L-B
College of Forestry, Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China.
Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
Insect Mol Biol. 2018 Apr;27(2):188-197. doi: 10.1111/imb.12365. Epub 2017 Nov 29.
Apolygus lucorum is the predominant pest of Bacillus thuringiensis (Bt) cotton in China. 20-hydroxyecdysone (20E) plays a key role in the reproduction of this insect. To better understand the mechanism underlying 20E-regulated reproduction, the nuclear hormone receptor E75 isoform-A of Ap. lucorum (Al-E75A) was cloned and its expression analysed. A 2241-bp sequence of Al-E75A cDNA encoded an open reading frame of a polypeptide with a predicted molecular mass of 69.04 kDa. Al-E75A mRNA was detected in female adult stages of Ap. lucorum with peak expression in 7-day-old animals. Al-E75A was also expressed in several tissues, particularly in the fat body and ovary. A 3.2 kb Al-E75A mRNA was detected in all tissues by Northern blot. The fecundity and longevity were significantly decreased in female adults treated with Al-E75A small interfering RNA. The rates of egg incubation rates were considerably lower in the RNA interference-treated animals compared to the untreated controls. In order to investigate the molecular mechanism underlying the effects described above, vitellogenin (Al-Vg) was selected for further investigation. The expression pattern of Al-Vg was similar to that of Al-E75A and was up-regulated by 20E. After knockdown of Al-E75A, the expression profile of Al-Vg and the protein levels were down-regulated. These findings suggest that Al-E75A plays a crucial role in the regulation of Al-Vg expression in Ap. lucorum.
绿盲蝽是中国转苏云金芽孢杆菌(Bt)基因棉花的主要害虫。20-羟基蜕皮酮(20E)在这种昆虫的繁殖中起关键作用。为了更好地理解20E调控繁殖的潜在机制,克隆了绿盲蝽的核激素受体E75亚型A(Al-E75A)并分析了其表达情况。Al-E75A cDNA的一个2241 bp序列编码了一个多肽的开放阅读框,预测分子量为69.04 kDa。在绿盲蝽雌成虫阶段检测到Al-E75A mRNA,在7日龄动物中表达达到峰值。Al-E75A也在多个组织中表达,尤其是在脂肪体和卵巢中。通过Northern印迹在所有组织中检测到一个3.2 kb的Al-E75A mRNA。用Al-E75A小干扰RNA处理的雌成虫的繁殖力和寿命显著降低。与未处理的对照相比,RNA干扰处理的动物的卵孵化率相当低。为了研究上述效应的分子机制,选择卵黄原蛋白(Al-Vg)进行进一步研究。Al-Vg的表达模式与Al-E75A相似,并且被20E上调。敲低Al-E75A后,Al-Vg的表达谱和蛋白水平下调。这些发现表明Al-E75A在绿盲蝽Al-Vg表达的调控中起关键作用。