Pan Yiou, Peng Tianfei, Gao Xiwu, Zhang Lei, Yang Chen, Xi Jinghui, Xin Xuecheng, Bi Rui, Shang Qingli
College of Plant Science, Jilin University, Changchun 130062, PR China.
Department of Entomology, China Agricultural University, Beijing 100193, PR China.
Comp Biochem Physiol Part D Genomics Proteomics. 2015 Mar;13:10-5. doi: 10.1016/j.cbd.2014.11.001. Epub 2014 Dec 5.
A thiamethoxam-resistant strain of cotton aphid (ThR) strain displayed a 19.35-fold greater resistance to thiamethoxam compared to a susceptible cotton aphid (SS) strain. Solexa sequencing technology was used to investigate differentially expressed genes (DEGs) in cotton aphids in the context of thiamethoxam resistance. A total of 22,569,311 and 21,317,732 clean reads were obtained from the ThR and SS transcriptomes, respectively, and assembled into 35,222 non-redundant (Nr) consensus sequences. The expression of 620 unigenes changed significantly in the ThR libraries compared to the SS strain; 349 genes were up-regulated, and 271 genes were down-regulated (P≤0.001). Expression levels of ribosomal proteins, ATP synthase, cytochrome c oxidase, ecdysteroid UDP-glucosyltransferase and esterase were up-regulated significantly in the ThR strain compared to the SS strain. The genes of cuticle proteins, salivary proteins, and fibroin heavy chain decreased dramatically. One nicotinic acetylcholine receptor (nAChR) α subunit was down-regulated in the ThR strain. The expression levels of 10 differentially expressed unigenes were confirmed using real-time RT-PCR, and the observed trends in gene expression matched the Solexa expression profiles. Specific single-nucleotide polymorphisms (SNPs) in nAChRs that cause amino acid substitution were found from the ThR and SS stains respectively. These data illustrate that genetic changes in nAChR genes and up-regulated ribosomal proteins, ecdysteroid UDP-glucosyltransferase, cytochrome c oxidase, esterase and peroxidase may confer the tolerance of resistant cotton aphids to thiamethoxam.
与敏感棉蚜（SS）品系相比，噻虫嗪抗性棉蚜品系（ThR）对噻虫嗪的抗性高19.35倍。利用Solexa测序技术研究棉蚜在噻虫嗪抗性背景下的差异表达基因（DEG）。分别从ThR和SS转录组中获得了22,569,311和21,317,732条clean reads，并组装成35,222条非冗余（Nr）共有序列。与SS品系相比，ThR文库中有620个单基因的表达发生了显著变化；349个基因上调，271个基因下调（P≤0.001）。与SS品系相比，ThR品系中核糖体蛋白、ATP合酶、细胞色素c氧化酶、蜕皮甾体UDP-葡萄糖基转移酶和酯酶的表达水平显著上调。表皮蛋白、唾液蛋白和丝素重链的基因显著减少。ThR品系中一个烟碱型乙酰胆碱受体（nAChR）α亚基下调。利用实时RT-PCR对10个差异表达单基因的表达水平进行了验证，观察到的基因表达趋势与Solexa表达谱相匹配。分别从ThR和SS品系中发现了导致氨基酸替换的nAChR中的特异性单核苷酸多态性（SNP）。这些数据表明，nAChR基因的遗传变化以及核糖体蛋白、蜕皮甾体UDP-葡萄糖基转移酶、细胞色素c氧化酶、酯酶和过氧化物酶的上调可能赋予抗性棉蚜对噻虫嗪的耐受性。
