Department of Agroecology, Aarhus University, Slagelse, Denmark.
Department of Agricultural Technology, Alexander Technological Educational Institute of Thessaloniki, Thessaloniki, Greece.
Insect Sci. 2019 Apr;26(2):274-282. doi: 10.1111/1744-7917.12539. Epub 2017 Nov 21.
When investigating insecticide resistance of pest insects, for example, the pollen beetle Meligethes aeneus, it is relevant to differentiate toxicological and molecular genetic data between male and female specimens. A molecular sex determination method would allow resistance testing to be run without prior sorting of the samples. A one-step quantitative RT-PCR method for quantification of the yolk protein vitellogenin expression in the pollen beetle was established. The expression level of vitellogenin relative to tubulin was determined. Pollen beetles were tested at different time points during their development to determine if vitellogenin is a reliable molecular marker for detection of sexually mature females. The differentiation between females and males by relative expression of vitellogenin to tubulin is conditional regarding the life cycle. Sexually mature females and males could easily be distinguished, whereas immature specimens could not be seperated. Vitellogenin expression is a successful marker for identification of sexually mature pollen beetles. Females from the spring populations showed vitellogenin expression when the temperature was above 10.2°C. Further, detailed observations of vitellogenin throughout the spring indicated a strong relationship between daily temperatures and vitellogenin expression, which is an indicator of oviposition ability.
在研究害虫的杀虫剂抗性时,例如花粉象甲(Meligethes aeneus),区分雄性和雌性标本的毒理学和分子遗传学数据是很重要的。一种分子性别鉴定方法可以在不事先对样本进行分类的情况下进行抗性测试。本文建立了一种一步定量 RT-PCR 方法,用于定量花粉象甲卵黄蛋白原表达。测定了卵黄蛋白原相对于微管蛋白的表达水平。在花粉象甲发育的不同时间点进行测试,以确定卵黄蛋白原是否是检测性成熟雌性的可靠分子标记。通过相对表达量来区分雌雄取决于生命周期。性成熟的雌性和雄性很容易区分,而未成熟的标本则无法区分。卵黄蛋白原的表达是鉴定性成熟花粉象甲的成功标记。春季种群中的雌性在温度高于 10.2°C 时表现出卵黄蛋白原表达。此外,对整个春季的卵黄蛋白原的详细观察表明,每日温度与卵黄蛋白原表达之间存在很强的关系,这是产卵能力的一个指标。