Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
J Insect Physiol. 2014 Jan;60:145-52. doi: 10.1016/j.jinsphys.2013.11.011. Epub 2013 Dec 8.
The evolution of Bacillus thuringiensis (Bt) resistance provides a useful pathway for the study of fitness trade-offs associated with stress adaptation. In a previous study, we used cDNA-amplification fragment length polymorphism (cDNA-ALFP) analysis to identify differentially expressed genes (DEGs) in Cry1Ac-susceptible (96S) and -resistant (Bt-R) Helicoverpa armigera larvae. Among these DEGs, a death-associated LIM-only protein (HaDALP) was identified. In the current study, the full-length cDNA encoding HaDALP was cloned using rapid amplification of cDNA ends according to an expressed sequence tag derived from the previous cDNA-ALFP analysis. HaDALP expression patterns indicated that this gene was differentially expressed in tissues and stages and was highest in the midgut and epidermis of the 5th instar larvae. It is up-regulated in Cry1Ac-resistant H. armigera larvae. Fitness parameters, such as larval and pupal weight, pupal duration, and survival rate, which are the most sensitive indicators, were evaluated after HaDALP knockdown or feeding of the HaDALP protein in vivo. Our findings suggested that up-regulation of HaDALP might be related to Cry1Ac resistance and an adaption to Bt toxins.
苏云金芽孢杆菌（Bt）抗性的进化为研究与应激适应相关的适应性权衡提供了一条有用的途径。在之前的一项研究中，我们使用 cDNA 扩增片段长度多态性（cDNA-ALFP）分析鉴定了对 Cry1Ac 敏感（96S）和抗性（Bt-R）棉铃虫幼虫差异表达的基因（DEGs）。在这些 DEGs 中，鉴定到一个死亡相关的 LIM 只有蛋白（HaDALP）。在本研究中，根据先前的 cDNA-ALFP 分析中源自表达序列标签的序列，使用快速扩增 cDNA 末端克隆编码 HaDALP 的全长 cDNA。HaDALP 的表达模式表明，该基因在组织和发育阶段存在差异表达，在 5 龄幼虫的中肠和表皮中表达最高。在 Cry1Ac 抗性棉铃虫幼虫中，它被上调表达。在体内敲低 HaDALP 或喂食 HaDALP 蛋白后，评估了幼虫和蛹重、蛹期和存活率等最敏感的适应参数。我们的研究结果表明，HaDALP 的上调可能与 Cry1Ac 抗性和对 Bt 毒素的适应有关。
