›› 2017, Vol. 60 ›› Issue (6): 691-698.doi: 10.16380/j.kcxb.2017.06.009

• 研究论文 • 上一篇    下一篇

放置时间和条件对空僵蚜中蚜虫和寄生蜂DNA检出率的影响

姚志文1,2, 杨帆2, 吴月坤1,2, 魏洪义1,*, 陆宴辉2,*   

  1. (1. 江西农业大学农学院, 南昌 330045; 2. 中国农业科学院植保保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193)
  • 出版日期:2017-06-20 发布日期:2017-06-20

Influences of storage time and conditions on the DNA detection rates of aphids and parasitoids from empty aphid mummies

YAO Zhi-Wen1,2, YANG Fan2, WU Yue-Kun1,2, WEI Hong-Yi1,*, LU Yan-Hui2,*   

  1. (1. College of Agronomy, Jiangxi Agricultural University, Nanchang 330045, China; 2. State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China)
  • Online:2017-06-20 Published:2017-06-20

摘要: 【目的】本研究以麦长管蚜Sitobion avenae (Fabricius)、麦二叉蚜Schizaphis graminum (Rondani)及初级寄生蜂烟蚜茧蜂Aphidius gifuensis (Ashmead)和重寄生蜂蚜虫宽缘金小蜂Pachyneuron aphidis (Bouché)为研究对象,分析空僵蚜放置时间和条件对蚜虫、寄生蜂DNA检出率的影响。【方法】将空僵蚜放置不同时间(4, 8, 12, 16, 20和24 d),纯水浸泡(4 h)或不浸泡并放置在不同温度(16℃, 24℃和32℃)和相对湿度(RH 40%和80%)组合条件下,定期取样并提取DNA,利用物种特异性引物进行扩增,检测空僵蚜中残留的蚜虫和寄生蜂DNA信息。【结果】随着空僵蚜放置时间的延长,两种寄生蜂的DNA检出率均逐步降低,在寄生蜂出蜂24 d后空僵蚜中,烟蚜茧蜂DNA在麦长管蚜和麦二叉蚜空僵蚜中的检出率分别为66.7%和86.7%,蚜虫宽缘金小蜂DNA在两种蚜虫中的检出率则都为66.7%。在浸泡处理以及不同温度和相对湿度条件下,寄生蜂出蜂后24 d的空僵蚜中两种蚜虫和烟蚜茧蜂DNA检出率在76.7%~100%之间。【结论】本研究结果验证了空僵蚜中残留蚜虫与寄生蜂DNA的稳定性,说明空僵蚜分子检测技术可有效应用于蚜虫寄生蜂食物关系的解析。

关键词: 麦长管蚜, 麦二叉蚜, 烟蚜茧蜂, 蚜虫宽缘金小蜂, 空僵蚜, 分子检测, 放置时间, 放置条件

Abstract: 【Aim】 This study aims to study the influences of storage time and conditions on the DNA detection rates of the common aphid species Sitobion avenae (Fabricius) and Schizaphis graminum (Rondani), the primary parasitoid Aphidius gifuensis (Ashmead) and the hyperparasitoid Pachyneuron aphidis (Bouché). 【Methods】 The empty aphid mummies were stored for different periods (4, 8, 12, 16, 20 and 24 d after parasitoid emergence), soaked in water for 4 h or unsoaked and kept at different temperature (16, 24 and 32℃) and humidity (RH 40% and 80%) combinations. The samples were collected at different time and stored for DNA extraction. The extracted DNA from the empty aphid mummies were detected using the species-specific primers. 【Results】 The results showed that the DNA detection rates of both parasitoid species decreased gradually with the storage time. In the empty mummies collected 24 d after parasitoid emergence, the A. gifuensis DNA was detected from 66.7% of S. avenae mummies and 86.7% of S. graminum mummies, respectively, and the P. aphidis DNA was detected from 66.7% of both aphid mummies. The DNA detection rates of S. avenae and S. graminum aphids and A. gifuensis wasps among soaked/unsoaked treatments or other different temperature and relative humidity combination treatments were 76.7%-100% from the empty aphid mummies collected 24 d after parasitoid emergence. 【Conclusion】 The results verified the stability of parasitoid DNA residues in wheat aphid mummy shells and the feasibility of DNA detection technique for empty aphid mummies. This offers a more convenient method to explore the aphid-parasitoid interactions.

Key words: Sitobion avenae, Schizaphis graminum, Aphidius gifuensis, Pachyneuron aphidis, empty aphid mummy, DNA detection, storage time, storage conditions