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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2013, Vol. 39 Issue (5): 473-590    DOI: 10.3785/j.issn.1008-9209.2013.02.192
生物科学与技术     
褐飞虱体内Himetobi P病毒的检测及组织定位
黄玉吉1，2，  陈斌1*， 张传溪2*
（1.云南农业大学植物保护学院， 昆明 650201；2.浙江大学昆虫科学研究所， 杭州 310058）
Detection and tissue-localization of Himetobi P virus in Nilaparvata lugens (Stal)
HUANG Yuji1，2, CHEN Bin1*, ZHANG Chuanxi2*
(1. College of Plant Protection, Yunnan Agricultural University, Kunming 650201, China; 2. Institute of Insect Science, Zhejiang University, Hangzhou 310058, China)
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摘要： 通过对Himetobi P病毒（HiPV）在褐飞虱［Nilaparvata lugens (Stl)］不同地理种群之间的差异、不同发育阶段的感染水平及不同器官组织的感染情况等方面的研究，为进一步研究HiPV与褐飞虱的互作及HiPV的应用奠定基础。对采自亚洲各地的10个褐飞虱种群进行实时荧光定量聚合酶链反应（real-time fluorescent quantitation-polymerase chain reaction， FQ-PCR)检测，发现所有检测的种群都携带有HiPV。根据HiPV衣壳蛋白编码序列进行聚类分析表明，不同地理来源宿主的病毒聚类与采集的地理远近并不一致。该病毒对褐飞虱没有明显的致病作用，可能是一种共生病毒。FQ-PCR分析表明，褐飞虱体内HiPV的相对含量随褐飞虱龄期的增大而缓慢增加，在成虫期达到高峰，且成虫中雄虫的带毒量高于雌虫。以FQ-PCR和免疫组织化学对宿主不同组织的病毒感染水平进行检测表明，HiPV在宿主的中肠后端部分感染水平最高，马氏管次之，而卵巢、精巢、唾液腺、脂肪体中含量较低。
Abstract: The brown planthopper (BPH), Nilaparvata lugens (Stl), is a serious insect pest of rice plants causing sucking damage and transmitting rice viruses, such as rice ragged stunt virus and rice grassy stunt virus. BPH has caused serious yield losses in Asia in recent decades. Himetobi P virus (HiPV) is a single-stranded RNA virus infecting rice planthoppers. Previous studies showed that the genome of this virus in Laodelphax striatellus is consisted of 9275 nucleotides encoding two large open reading frames (ORFs). It is not clear so far the diversity of HiPV regarding strain from various geographic host populations, and its infection status in different developmental stages and tissues of N. lugens. HiPVs in 10 BPH populations collected from  different areas in Asia, including five populations from China, three populations from Philippine, one population from Malaysia and one population from Laos, were detected by real-time  fluorescent quantitation-polymerase chain reaction  （FQ-PCR）.  All populations were found to be infected by HiPV, suggesting that BPH populations were extensively infected by HiPV. In phylogenetic analysis based on the viral capsid protein sequences, the 10 virus strains were clustered into four groups with less relevance to the host geographic locations.
The virus was  not evidently pathogenic to BPH, suggesting that it may be a symbiont of the insect. Real-time PCR detections showed that the relative amount of HiPV in BPH increased gradually along with nymph growth and increased sharply after adult emergence. More male adults were infected with HiPV than female adults.
Real-time PCR and immunohistochemical staining results demonstrated that the post part of the BPH midgut had  the highest HiPV concentration, followed by Malpighian tubules. Ovaries, spermaries, salivary glands and fat bodies were also infected with HiPV, but at a relatively lower  level. This suggested that the circumstance of digestive tract might be more appropriate for virus multiplication, which was  accordant with the fact that HiPV might be also related with its oral transmission. In conclusion, our results showed that BPH populations were extensively infected by HiPV and the post part of the midgut in the adult stage was the mainly infected tissues. The virus seems not evidently pathogenic to BPH, however, its biological functions in the symbiosis, i.e., its effects on host development and reproduction and on the transmission of the rice viruses, are not clear yet. Our results are expected to serve as a base for further researches on the relationship between Himetobi P virus and BPH host and developing of HiPV-based novel methods for the control of BPH.  
出版日期: 2013-09-20
CLC:  Q 965.8  
基金资助:

国家重点基础研究发展计划(973计划)资助项目（2010CB126200）；国家自然科学基金资助项目（31070136）；云南省科技创新团队计划资助项目（2011HC005）；云南省高校科技创新团队支持计划资助项目［云教科（2011）14号］。
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陈斌1*
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张传溪2*

引用本文:

黄玉吉1，2， 陈斌1*， 张传溪2*. 褐飞虱体内Himetobi P病毒的检测及组织定位[J]. 浙江大学学报(农业与生命科学版), 2013, 39(5): 473-590.

HUANG Yuji1，2, CHEN Bin1*, ZHANG Chuanxi2*. Detection and tissue-localization of Himetobi P virus in Nilaparvata lugens (Stal). Journal of Zhejiang University (Agriculture and Life Sciences), 2013, 39(5): 473-590.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2013.02.192    或    http://www.zjujournals.com/agr/CN/Y2013/V39/I5/473

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