Detection of Yeast-like Symbionts in Brown Planthopper Reared on Different Resistant Rice Varieties Combining DGGE and Absolute Quantitative Real-Time PCR.
Detection of Yeast-like Symbionts in Brown Planthopper Reared on Different Resistant Rice Varieties Combining DGGE and Absolute Quantitative Real-Time PCR.
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结合 DGGE 和绝对定量实时 PCR 检测不同抗性水稻品种褐飞虱中的类酵母共生菌
DOI：
10.3390/insects13010085
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发表时间：
2022-01-13
期刊：
影响因子：
3
通讯作者：
Yu X
中科院分区：
文献类型：
--
作者：
Lai C;Hou Y;Hao P;Pang K;Yu X
褐飞虱是水稻生产中的一种重要害虫，给水稻生产造成巨大损失。推广和利用抗虫水稻品种是防治褐飞虱的重要途径。然而，在实践中，褐飞虱在几代内就能适应抗病水稻。内共生菌可能是褐飞虱对抗病水稻快速适应的原因之一。褐飞虱的腹部含有酵母样共生菌(YLS)，YLS是BPH的营养、发育和繁殖所必需的。我们以前的报告表明，在褐飞虱中发现的YLS群落中，子囊菌共生体、类毕赤酵母类共生体和类假丝酵母类共生体是YLS的三个优势种群。本研究采用PCR-DGGE和实时荧光定量聚合酶链式反应技术，对褐飞虱3个显性YLS在不同虫龄和不同抗性水稻品种上的变异进行了检测。结果表明，在感病品种TN1上饲养褐飞虱，从1龄到成虫，幼虫总数逐渐增加，5龄若虫逐渐减少。抗病品种Mudgo、ASD7和RH对褐飞虱第1代和第2代的3种优势YLS的抑制作用更为显著。3个优势YL的数量均从第三代BPH中恢复。子囊菌共生菌是三种菌株中最具优势的菌株。褐飞虱是亚洲水稻的一种重要害虫。类酵母共生菌(YLS)是与褐飞虱的发生发展和褐飞虱对抗性植物的毒力适应机制密切相关的内生共生菌。在这项研究中，我们使用半定量DGGE和绝对定量实时聚合酶链式反应(QPCR)来定量检测在若虫期和新出现的雌性成虫中感染BPH的三种YLS菌株(子囊菌共生菌、毕赤酵母样共生菌和假丝酵母样共生菌)的数量。3种幼虫的数量均随若虫龄期的发育而增加，在4龄时达到峰值，5龄时显著下降。然而，在初露头角的成年雌性体内，YLS的数量急剧恢复。此外，我们估计了YLS在接种到抗病品种(Mudgo、ASD7和RH)上后长达8代的数量，以重新将YLS的动态与褐飞虱的适合度联系起来。抗病品种在第2代检测到最小的YLS数，从第3代开始逐渐增加。此外，YLS的子囊菌共生体被发现是在BPH所有发育阶段所测试的三个YLS菌株中最丰富的。
The brown planthopper (BPH) is an important pest that causes huge losses in rice production. The promotion and use of insect-resistant rice varieties is an important way to control BPH. However, in practice, BPH can adapt to resistant rice within several generations. Endosymbionts may be one of the reasons for the rapid adaptation of BPH to resistant rice. The BPH harbor yeast-like symbionts (YLS) in their abdomen, and YLS are essential for the nutrition, development, and reproduction of BPH. Our previous report showed that among the YLS communities detected in BPH, Ascomycetes symbionts, Pichia-like symbionts, and Candida-like symbionts were the three dominant populations of YLS. In this study, PCR-DGGE and absolute quantitative real-time PCR were used to detect the variations of three dominant YLS in BPH across different nymph ages and on different resistant rice varieties. The results showed that the total number of YLS gradually increased from the first instar to adulthood, but decreased in the fifth instar nymph, when BPH were reared on the susceptible rice variety TN1. The rice-resistant varieties, Mudgo, ASD7, and RH have more significant inhibitory effects on the three dominant YLS in the first and second generations of BPH. However, the numbers of the three dominant YLS were all recovered from the third generation of BPH. Ascomycetes symbionts were the most dominant strain among the three YLS. The brown planthopper (BPH), Nilaparvata lugens, is a serious pest of rice throughout Asia. Yeast-like symbionts (YLS) are endosymbionts closely linked with the development of BPH and the adapted mechanism of BPH virulence to resistant plants. In this study, we used semi-quantitative DGGE and absolute quantitative real-time PCR (qPCR) to quantify the number of the three YLS strains (Ascomycetes symbionts, Pichia-like symbionts, and Candida-like symbionts) that typically infect BPH in the nymphal stages and in newly emerged female adults. The quantities of each of the three YLS assessed increased in tandem with the developing nymphal instar stages, peaking at the fourth instar stage, and then declined significantly at the fifth instar stage. However, the amount of YLS present recovered sharply within the emerging adult females. Additionally, we estimated the quantities of YLS for up to eight generations after their inoculation onto resistant cultivars (Mudgo, ASD7, and RH) to reassociate the dynamics of YLS with the fitness of BPH. The minimum number of each YLS was detected in the second generation and gradually increased from the third generation with regard to resistant rice varieties. In addition, the Ascomycetes symbionts of YLS were found to be the most abundant of the three YLS strains tested for all of the development stages of BPH.
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