褐飞虱特异引物的设计和评估

Design and evaluation of the species-specific primers of the brown planthopper,Nilaparvata lugens(Stl)(Homoptera:Delphacidae)

研究稻田生态系统中天敌对褐飞虱Nilaparvata lugens(Stl)的捕食非常重要,但很困难。本研究设计褐飞虱的PCR特异引物,并对利用从稻田66种稻田常见节肢动物中提取的DNA进行了特异性验证,得到褐飞虱的特异引物(Lug-F1a/Lug-R1),在实验室以拟环纹豹蛛Pardosa pseudoannulata-褐飞虱为模式设置试验,分析消化时间、温度、天敌取食量和天敌性别对褐飞虱特异引物的阳性反应率的影响。结果发现:捕食者性别对检测阳性反应没有显著影响,而消化时间、温度和取食量均对引物的检测有显著影响。其中,检测阳性率随消化时间增加而下降,两者呈指数函数关系。褐飞虱特异引物的设计和评估,对下一步在田间应用该引物奠定了基础,为研究褐飞虱与天敌之间的关系提供了新的工具和思路。

Predation of the brown planthopper（BPH）,Nilaparvata lugens（Stl） by its predators is particularly important in rice field ecosystem,but its studying is rather difficult.In this study,PCR cross-amplification was initially conducted on DNA extracted from 66 species of arthropods commonly found in rice fields.Only Lug-F1a/Lug-R1 was specific and efficient enough to detect species-specific DNA of N.lugens in the gut of wolf spider Pardosa pseudoannulata.Then,we assessed how digestion time,temperature,feeding amount of predators and the sex of predators can influence the effectiveness of detecting BPHs in spider guts using Lug-F1a/Lug-R1.The results indicated that digestion time,temperature and feeding amount significantly affected the detection of the primers,while the sex of predators had no significant influence.The detection was significantly and negatively correlated with digestion time and temperature,but positively with the feeding amount.Thus the species-specific primers of N.lugens had been successfully identified,which could be effectively used in rice fields.These primers under the conditions tested here provide a new detection tool efficient for studying the relationship between N.lugens and its predators.