绿盲蝽卵黄原蛋白(Vg)基因片段克隆及不同生境表达量分析

Vitellogenin(Vg)Gene Fragment from Apolyguslucorum(Meyer-Dür.):Cloning and Expression Levels in Different Habitats

为了探讨不同果园生境对绿盲蝽生殖的影响,从分子角度确定绿盲蝽适合的产卵生境,本研究首先采用绿盲蝽专用诱芯调查了枣园、葡萄园的绿盲蝽发生量,然后通过比较半翅目,直翅目,膜翅目,鳞翅目等昆虫cDNA同源序列,根据基因保守序列设计一对简并性引物,克隆绿盲蝽卵黄原蛋白(Vg)基因片段,采用RT-PCR的方法检测了2种生境中Vg基因相对表达量。结果显示,枣园绿盲蝽的发生量始终高于葡萄园,枣园有4个发生高峰,而葡萄园有2个。基因克隆获得一段大小为732 bp的基因片段,编码244个氨基酸;该基因序列与其他蝽类昆虫Vg基因氨基酸序列的同源性达80%以上;系统进化分析表明,绿盲蝽Vg基因与赤须盲蝽、烟盲蝽及黑肩绿盲蝽的亲缘关系最近,克隆的基因片段为绿盲蝽Vg基因片段,并命名为AL-Vg。不同生境Vg基因表达量比较可知,枣园的表达量低于葡萄园,结合发生量表明枣园生境更适合绿盲蝽生殖产卵。

To explore the effects of different orchard habitats on the reproduction of Apolyguslucorum(Meyer-Dür.),and determine the suitable spawning habitat of A.lucoruminthe orchard from a molecular perspective,a special lure was used to investigate the occurrence of A.lucorumin in jujube gardens and vineyards.By comparing the cDNA homologous sequences of Hemiptera,Orthoptera,Hymenoptera,Lepidoptera,etc.,a pair of degenerate primers was designed according to gene conservation to clone vitellogenin(Vg)gene fragment,and the expressions of Vg gene in two habitats were detected by RT-PCR.The results showed that the occurrence of A.lucorum in jujube gardens was constantly higher than that in vineyards.There were three peaks in jujube gardens and two in vineyards.By gene cloning,a 732 bp gene sequence fragment was obtained which encoded 244 amino acids,and the homology of the gene sequence with other Vg gene amino acids sequences was above 80%.The cloned gene fragment was a A.lucoruminthe Vg gene fragment and was named AL-Vg.Comparison of Vg gene expression in different habitats shows that the jujube garden habitat is more suitable for the reproduction and spawning of A.lucorum.