Objective: To investigate the genes involved in Wolbachia-induced cytoplasmic incompatibility among three natural populations of Culex pipiens pallens in eastern China, so as to provide insights into the development of preventive and control measures for mosquito-borne diseases based on Wolbachia.
Methods: The cytoplasmic incompatibility was tested among three natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province and Tangkou of Shandong Province using reciprocal crosses. Wolbachia infection was detected in C. pipiens pallens using a PCR assay, and the expression of Wolbachia wsp and WD0513 genes was quantified using a fluorescent quantitative real-time PCR (qPCR) assay.
Results: Bidirectional compatibility was found between the natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province (t = 0.57 and 0.15, both P values > 0.05), while bidirectional incompatibility was seen between the natural populations of C. pipiens pallens collected from Tangkou of Shandong Province and Wuxi of Jiangsu Province (t = 63.81 and 43.51, both P values < 0.01), and between the natural populations of C. pipiens pallens collected from Nanjing of Jiangsu Province and Tangkou of Shandong Province (t = 39.62 and 43.12, both P values < 0.01). Wolbachia wsp gene was amplified in all three natural populations of C. pipiens pallens, and qPCR assay detected no significant difference in the Wolbachia wsp gene expression among the three natural populations of C. pipiens pallens (F = 2.15, P > 0.05). In addition, there was no significant difference in the WD0513 gene expression between the natural populations of C. pipiens pallens collected from Tangkou of Shandong Province and Nanjing of Jiangsu Province (q = 8.42, P < 0.05) or between the natural populations of C. pipiens pallens collected from Tangkou of Shandong Province and Wuxi of Jiangsu Province (q = 7.84, P < 0.05); however, there was a significant difference detected in the WD0513 gene expression between the natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province (q = 0.40, P > 0.05).
Conclusions: Different Wolbachia numbers are detected in natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province and Tangkou of Shandong Province, and WD0513 gene may be involved in the Wolbachia-induced cytoplasmic incompatibility among three natural populations of C. pipiens pallens.
［摘要］ 目的探讨我国东部地区3个地理种群淡色库蚊由沃尔巴克氏体导致细胞质不亲和的相关基因, 为制定基于 沃尔巴克氏体的蚊媒病防制措施提供参考。方法采用正反交实验确定江苏南京、江苏无锡和山东唐口3个淡色库蚊 自然种群间细胞质不亲和, 应用PCR技术检测淡色库蚊沃尔巴克氏体感染情况, 采用荧光定量PCR法检测沃尔巴克氏 体wsp 基因和沃尔巴克氏体基因组中Octomom增殖区域WD0513 基因表达水平。结果江苏南京与无锡淡色库蚊自然 种群双向亲和 (t = 0.57、0.15, P 均> 0.05), 山东唐口和江苏无锡淡色库蚊自然种群双向不亲和 (t = 63.81、43.51, P 均< 0.01), 江苏南京和山东唐口淡色库蚊自然种群双向不亲和 (t = 39.62、43.12, P 均< 0.01) 。3个地理种群淡色库蚊基因组 均扩增出wsp 基因条带。荧光定量PCR检测发现, 3个自然种群淡色库蚊感染的沃尔巴克氏体wsp 基因表达水平不同, 但 差异无统计学意义 (F = 2.15, P > 0.05) 。山东唐口与江苏南京、山东唐口与江苏无锡淡色库蚊自然种群WD0513 基因表 达水平间差异均有统计学意义 (q = 8.42、7.84, P 均< 0.05), 但江苏南京与江苏无锡淡色库蚊自然种群WD0513 基因表达 水平间差异无统计学意义 (q = 0.40, P > 0.05) 。结论江苏南京、江苏无锡和山东唐口3个地理种群淡色库蚊所感染沃 尔巴克氏体数量不同, 淡色库蚊自然种群感染沃尔巴克氏体导致的细胞质不亲和可能与WD0513 基因表达量相关。.
Keywords: Culex pipiens pallens; Cytoplasmic incompatibility; WD0513 gene; Wolbachia.