Liang Guang Hong, Jang Eric B, Heller Wade P, Chang Chiou Ling, Chen Jia Hua, Zhang Fei Ping, Geib Scott M
Fujian Agriculture and Forestry University, College of Forestry, Fuzhou, China.
USDA-ARS, DKI-PBARC, Hilo, USA.
Pest Manag Sci. 2015 Dec;71(12):1666-74. doi: 10.1002/ps.3976. Epub 2015 Feb 4.
Parasitism detection and species identification are necessary in fruit fly biological control. Currently, release of mass-reared Fopius arisanus is practiced worldwide, as it is effective in controlling Bactrocera dorsalis and Ceratitis capitata. To detect and assess parasitism in parasitoid mass-rearing colonies and parasitism levels in field populations across all life stages of hosts, the development of a rapid, specific and sensitive method is important.
A species-specific probe was designed for F. arisanus, as well as a universal tephritid probe. Utilizing rapid DNA extraction techniques coupled with quantitative-PCR, a simple and fast assay has been developed to detect parasitism of F. arisanus that is sensitive enough to detect the parasitoid across all developmental stages, including a single egg per host egg or 0.25 ng of parasitoid DNA in 40 ng of host DNA. The qPCR methods also detect a higher parasitism rate when compared with rearing-based methods where parasitism rate is based on wasp emergence and where unemerged wasps are not included.
This method is a rapid, sensitive and specific technique to determine the parasitism rate of F. arisanus across all life stages of B. dorsalis, which will be useful to predict parasitoid output from mass rearing and evaluate the outcome of pest suppression after mass release in the field.
在果蝇生物防治中,寄生虫检测和物种鉴定是必要的。目前,全球都在大规模释放人工饲养的阿里山潜蝇茧蜂,因为它对控制橘小实蝇和地中海实蝇有效。为了检测和评估寄生蜂大规模饲养群体中的寄生情况以及寄主所有生命阶段田间种群的寄生水平,开发一种快速、特异且灵敏的方法很重要。
设计了一种针对阿里山潜蝇茧蜂的物种特异性探针以及一种实蝇通用探针。利用快速DNA提取技术结合定量PCR,开发了一种简单快速的检测方法,用于检测阿里山潜蝇茧蜂的寄生情况,该方法灵敏度足以检测寄生蜂在所有发育阶段的情况,包括每个寄主卵中有单个卵或40纳克寄主DNA中有0.25纳克寄生蜂DNA。与基于饲养的方法相比,定量PCR方法检测到的寄生率更高,基于饲养的方法中寄生率是基于黄蜂羽化情况,且未羽化的黄蜂不计算在内。
该方法是一种快速、灵敏且特异的技术,可用于确定阿里山潜蝇茧蜂在橘小实蝇所有生命阶段的寄生率,这将有助于预测大规模饲养中寄生蜂的产量,并评估田间大规模释放后害虫抑制的效果。