为了明确羧酸酯酶(carboxylesterase,CarE)基因、谷胱甘肽S-转移酶(glutathione S-transfer-ases,GST)基因和过氧化氢酶(catalase,CAT)基因与柑橘全爪螨Panonychus citri对双甲脒抗性的关系,通过BLAST检索,从柑橘全爪螨转录组数据库中对这3种代谢酶抗性相关基因进行鉴定,并采用RPKM法对双甲脒抗性品系和敏感品系代谢抗性相关基因进行表达差异分析,对差异较大的基因作定量PCR检测。基因差异性分析发现,抗性品系中有9条CarE基因、12条GST基因及6条CAT基因表达量发生上调,13条CarE基因、12条GST基因和3条CAT基因表达量发生下调;Pc29773nrt、Pc17807nlg和Unigene31477为上调倍数最高的3个基因,其log2ratio(RS/SS)分别为12.95、10.81、10.01。定量分析显示,Pc29773nrt、Pc17807nlg和Unigene31477的上调倍数分别为3.72、2.03和3.09,Pc29773nrt和Unigene31477上调显著。研究表明柑橘全爪螨Pc29773nrt和Unigene31477上调与其对双甲脒的抗性相关。
To clarify the relationship between expression level of carboxylesterase (CarE) gene, gluta- thione S-transferase (GST) gene and catalase (CAT) gene in resistance of Panonychus citri, three kinds of resistance-related metabolic enzyme genes were identified based on BLAST, as well as differential ex- pression profiling analysis for both the amitraz-resistant strain and susceptible strain through RPKM meth- od, and the higher up-regulated genes were selected for real-time PCR validation. Gene expression differ- ence analysis results indicated that nine Care genes, 12 GST genes, and six CAT genes were up-regula- ted and 13 Care genes, 12 GST genes and three CAT genes were down-regulated in resistant strain. Pc29773nrt, Pc17807nlg and Unigene31477 were the three top up-regulated genes, log2 ratio (RS/SS)was 12.95, 10.81, 10.01, respectively. Further real-time PCR analysis indicated that Pc29773nrt, Pc17307nlg and Unigene31477 had a 3.72, 2.03 and 3.09 fold up-regulation respectively in resistant strain. Pc29773nrt and Unigene31477 were significantly up-regulated. The above results indicated that up-regulation of Pc29773nrt and Unigene31477 play an important role in amitraz resistance of P. cirri.