褐飞虱持续暴发成因——以江西泰和县为例

为了明确泰和县近年来晚稻田褐飞虱Nilaparvata lugens(Stl)连年暴发的规律。通过分析泰和县2008—2013年褐飞虱的田间调查及灯诱数据,并结合气象条件,研究了泰和县晚稻田褐飞虱暴发的主要影响因素。结果表明:(1)危害早稻田的褐飞虱以2、3代为主,其田间种群数量的多少主要受春季迁入虫量的限制,本地早稻收割后的滞留虫源对晚稻种群数量的影响很小。(2)危害晚稻田的褐飞虱以5、6代为主,后期迁入虫量、秋季温度以及晚稻田初期外来迁入虫量是影响晚稻田后期暴发的关键因子。(3)不同年份间晚稻田褐飞虱暴发类型不同。2008和2009年属后期迁入虫量与"暖秋"共同影响类型;2010年是"暖秋"影响类型;2011和2012年是后期迁入虫量影响类型;2013年是晚稻田早期迁入虫量积累影响类型。后期迁入虫量、秋季温度以及晚稻田早期迁入虫量是导致泰和县晚稻田褐飞虱连年暴发的主要原因。这些结果为褐飞虱暴发规律的阐明及可持续治理提供了理论依据和参考价值。     

褐飞虱体内能源物质含量与其迁飞关系研究初报

<正> 褐飞虱Nilaparvata lugens Stal进行远距离迁飞活动,势必消耗体内大量能源物质。本试验研究羽化幼嫩阶段的稻田褐飞虱(简称稻田类型〕和高山网捕获及当年测报灯下初见的褐飞虱(简称非稻田类型)胸部及整体能源物质的变化,分析能源物质与迁飞的关系。     

《褐飞虱研究与防治》

《褐飞虱研究与防治》一书 ,由陈遐年等教授主编 ,已于 2 0 0 3年 7月由中国农业出版社正式出版发行。本书全面、系统地论述了褐飞虱研究与治理领域的最新进展、研究方法和实际应用。全书分 :国内外褐飞虱研究进展、褐飞虱灾害的地理分布及其时空变化规律、褐飞虱的形态特征和稻田内近似种类的识别、褐飞虱的种群生物学特性、褐飞虱大发生的种群特征、褐飞虱大发生与稻田生物群落关系、诱发褐飞虱再猖獗的生理生态因素、褐飞虱的远距离迁飞行为及其生理生态机制、褐飞虱发生系统的混沌特性及预测预报和褐飞虱防治技术的研究与应用等 1 2章。…     

褐飞虱在香稻上的发生与产卵特征

研究褐飞虱NilaparvatalugensStl在香稻上的危害与产卵特征,结果表明香稻田中褐飞虱的自然种群密度基数远高于非香型稻。从7月22日至8月31日香稻田中褐飞虱的自然种群密度一直增长,在武香988和鄂香1号田间的若虫分别从7月22日的7头丛和3头丛上升至8月31日的126头丛和73头丛。香型稻田间的褐飞虱的自然种群密度增长慢于非香型稻田间。香稻与非香稻田间褐飞虱成虫出现的高峰日相差不大。香稻田中褐飞虱产卵株率高于非香型稻田,其中最高的为鄂香1号,达到74.27%。香稻田间株平均产卵块数几乎为非香型稻上的2倍,但是香稻上的卵死亡率(鄂香1号为39.24%)显著高于非香型稻上的(鄂中5号为27.58%,两优培九为27.21%),且产卵部位有一定差异。鄂香1号第Ⅲ叶鞘上卵块最多,占总数的60.98%,武香988的第Ⅱ和第Ⅳ叶鞘上产卵较多,第Ⅰ叶鞘上没有稻飞虱产卵。非香型稻鄂中5号上产卵最多的是第Ⅳ叶鞘,第Ⅰ和第Ⅱ叶鞘上没有褐飞虱产卵。两优培九除第Ⅰ和第Ⅴ叶鞘上没有褐飞虱产卵外,第Ⅱ、第Ⅲ和第Ⅳ叶鞘上均有褐飞虱产卵,而且卵量差异不大。     

氮肥影响节肢动物天敌对褐飞虱种群的自然控制作用

氮肥在增加粮食产量的同时也可能对整个农田生态系统产生负面影响。稻田过量施用氮肥后,会提高水稻对害虫的敏感性、改变害虫与天敌之间的关系,最终影响到天敌对害虫的自然控制功能,导致害虫大发生。为了合理、公正地评价施用氮肥对稻田节肢动物天敌对害虫自然控制能力的影响,探索性地应用笼罩的方法在菲律宾国际水稻研究所试验农场稻田中研究了害虫天敌在不同氮肥施用水平(0,100 kg N/hm~2和200 kg N/hm~2)稻田中对褐飞虱的捕食能力及自然控制作用。试验结果表明,旱季田间的捕食性天敌对褐飞虱若虫的捕食能力和主要天敌对褐飞虱种群的自然控制能力均随稻田氮肥施用量的增加而减弱。在雨季,虽然天敌对褐飞虱种群的自然控制能力也随稻田氮肥施用量的增加而减弱,但捕食性天敌对褐飞虱若虫捕食能力的差异不明显。本研究表明,天敌对褐飞虱自然控制能力的减弱是稻田过量施用氮肥后褐飞虱种群猖獗的主要原因之一。     

不同营养源对稻虱缨小蜂寿命及寄生能力的影响

研究了蜂蜜、玉米花粉、大豆花、褐飞虱蜜露和黄脊飞虱蜜露对稻虱缨小蜂寿命、寄生能力以及寄生行为对稻虱缨小蜂寿命和存活率的影响。结果表明,蜂蜜、玉米花粉、褐飞虱蜜露和大豆花均能明显延长稻虱缨小蜂的寿命。并且显著地提高了对褐飞虱卵的寄生能力,其中蜂蜜最有效,大豆花次之,玉米花粉和褐飞虱蜜露这两种营养源以玉米花粉(水和褐飞虱蜜露+水的形式)对提高稻虱缨小蜂寿命最有效,而单一玉米花粉、花粉液、褐飞虱蜜露稀释液和纯褐飞虱蜜露均不能延长稻虱缨小蜂寿命,黄脊飞虱蜜露对稻虱缨小蜂的寿命和寄生能力均无影响。寄生行为对稻虱缨小蜂的寿命基本无影响。但致使其40～48h内的存活率提高,此后的存活率降低较快,在稻田周围的作物和植被上调查到约10种飞虱,非稻田生境能为稻田寄生性天敌提供寄主和食物,是理想的庇护所,对保护和提高稻田天敌种群数量,提高稻田天敌的生物控制作用。     

水稻褐飞虱内生共生细菌Arsenophonus的鉴定和系统分析

利用16S rDNA通用引物扩增了水稻褐飞虱Nilaparvata lugens（Stål）体内共生细菌的序列,经克隆、测序和NCBI数据库比对,发现褐飞虱体内存在杀雄菌属Arsenophonus类共生细菌,系统发育上与粉虱科和木虱科体内的Arsenophonus属亲源关系较近。在褐飞虱体内该共生细菌具有两种长度不同的16S rDNA序列,分别为1 504 bp和547 bp,其中后者为前者中间缺失了957 bp,其余序列相同。通过重新设计两对引物进行扩增,进一步确认不同褐飞虱地理种群及寄主种群均存在两种片段。Arsenophonus特异的 23S rDNA引物的扩增结果表明,Arsenophonus存在于所有检测的褐飞虱种群中,但不存在于水稻寄主中。荧光定量PCR检测发现3个褐飞虱室内寄主种群Arsenophonus属共生细菌含量不同,其中TN1种群明显高于Mudgo种群和ASD7种群。此为水稻褐飞虱体内存在Arsenophonus属共生细菌的首次报道。     

综防区和非综区稻田生境中褐飞虱卵寄生蜂群落的比较

褐飞虱 ( Nilaparvatlugens)是一类水稻主要害虫。由于稻飞虱对抗虫水稻品种的适应 ,以及化学农药的毒副作用 ,其间歇性大发生的局面仍没有改变 ,因此 ,以保护利用自然天敌为主的生物防治方法日益受到重视。褐飞虱的天敌种类很多 [1～ 4 ] ,对褐飞虱的控制作用不是由某一种或几种完成的 ,而是褐飞虱天敌群落共同作用的结果 ,因而有必要在群落水平上研究褐飞虱天敌 ,了解天敌群落的结构和动态 ,从而使天敌对褐飞虱的控制作用更好地发挥。本文对综防区和非综防区稻田生境中稻飞虱卵寄生蜂群落的结构和动态进行了比较研究。1　材料与方法1 .1…     

褐飞虱蜕皮激素合成相关Halloween基因的克隆及功能分析

【目的】蜕皮激素(ecdysone)是调控昆虫生长发育和繁殖的重要激素,其合成主要由5个称为Halloween基因编码的细胞色素P450(spook/CYP307A1,Phantom/CYP306A1,disembodied/CYP302A1,shadow/CYP315A1和shade/CYP314A1)参与。褐飞虱Nilaparvata lugens系东南亚和东亚地区危害最为严重的水稻害虫之一,以蜕皮激素调控基因为靶标,应用RNAi技术防控褐飞虱正逐步成为研究热点。本研究旨在揭示褐飞虱体内Halloween基因的功能,为褐飞虱的防控和新农药的开发提供理论依据。【方法】本研究基于褐飞虱基因组和转录组数据,利用RT-PCR技术克隆了参与蜕皮激素合成的5个Halloween基因;利用生物信息学软件预测分析其编码蛋白的结构特性;利用MEGA5.0邻接法构建系统进化树;利用实时荧光定量PCR技术研究其时空表达特征;利用注射法RNAi技术分析Cyp314a1基因表达下调对褐飞虱生长发育和繁殖的影响。【结果】克隆获得褐飞虱的5个Halloween基因,即NlCyp307a1(Gen Bank登录号:KM217014.1),NlCyp306a1(Gen Bank登录号:KM217013.1),NlCyp302a1(Gen Bank登录号:KM216995.1),NlCyp315a1(Gen Bank登录号:KM216998.1)和NlCyp314a1(Gen Bank登录号:KU928172)。进化分析表明,这5个基因分属于CYP2和线粒体两大类群,并与各自的同源基因聚类在一起,说明Halloween基因和蜕皮激素合成途径在昆虫进化过程中十分保守。Nl CYP314A1具有5个P450的保守结构域和两个跨膜结构域,是典型的线粒体酶。qRT-PCR检测结果显示,5个Halloween基因的表达量在整个褐飞虱5龄若虫期内呈现波动状,且均在5龄若虫蜕皮后的24 h和60 h出现表达量峰值。NlCyp314a1在褐飞虱发育各个龄期均表达,且在成虫中表达量最高;NlCyp314a1在胸部表达量较头部和腹部高,其在脂肪体中的表达量最高,其次是在足、表皮、翅芽中,中肠中表达量最低。利用注射法RNAi对NlCyp314a1基因进行沉默后,NlCyp314a1的表达量在注射后第4天较对照极显著降低84.6%(P0.01),同时蜕皮激素通路下游的响应关键基因Nl FTZ-F1的表达量较对照极显著降低64.1%(P0.01)。表型观察发现,NlCyp314a1表达量的下调导致了褐飞虱蜕皮困难,出现畸形虫体,第7天死亡率大于95%,且NlCyp314a1表达量的下调导致了褐飞虱卵巢发育畸形。【结论】褐飞虱蜕皮激素合成相关的Halloween基因在进化上非常保守,Halloween基因功能的研究揭示了褐飞虱体内Halloween基因对于褐飞虱蜕皮和繁殖的重要性。其表达水平的降低导致了褐飞虱生长发育和繁殖受阻,提示褐飞虱Halloween基因可用于褐飞虱蜕皮和繁殖进程的调控,同样也为Halloween基因在褐飞虱防治方面的潜在应用提供了依据。     

利用分子探针法研究稻田蜘蛛集团对褐飞虱的捕食作用

为褐飞虱Nilaparvata lugens（Stal）的绿色防控提供理论参考依据,于2011年和2012年在调查了浙江富阳不同品种稻田褐飞虱和蜘蛛的发生密度后,利用荧光定量PCR分子探针技术分析了9科27种3 807头蜘蛛对褐飞虱的捕食作用。研究结果表明,在水稻Oryza sativa L.不同生育期,稻田总的褐飞虱和蜘蛛密度等参数值均随调查时间呈规律性变化,且在调查时间点间出现了显著性差异（P<0.05）;各科蜘蛛对褐飞虱的捕食阳性率均随水稻生育期的发展呈增加趋势,整体上,狼蛛科Lycosidae、皿蛛科Linyphiidae、球腹蛛科Theridiidae、肖蛸科Tetragnathidae、跳蛛科Salticidae和园蛛科Araneidae捕食率在DAT21,DAT35和DAT77,DAT91之间有显著性差异（P<0.05）;2012年两品种稻田的褐飞虱和蜘蛛密度等参数值均显著高于2011年的值,两年间汕优63（SY63）稻田的褐飞虱和蜘蛛密度均显著高于IR64的密度;2012年各科蜘蛛对褐飞虱的捕食阳性率显著高于2011年的值（P<0.05）,且狼蛛科和球腹蛛科对褐飞虱的捕食阳性率在两品种间存在显著性差异（P<0.05）;稻田4种蜘蛛优势种拟环纹豹蛛Pardosa pseudoannulata (Bosenberg & Strand)、八斑球腹蛛Theridion octomaculatum (Boes. et Str.）、食虫瘤胸蛛Oedothorax insecticeps (Boes. et str.)和锥腹肖蛸Tetragnatha maxillosa (Thoren)的捕食阳性率均随褐飞虱种群密度的增加而增加,该捕食功能反应曲线可用非线性模型P=aN/(1+bN)拟合;除锥腹肖蛸外,其它3种的捕食功能反应曲线均呈饱和状态;拟环纹豹蛛捕食褐飞虱的DNA残留量显著高于八斑球腹蛛、锥腹肖蛸和食虫瘤胸蛛的残留量（P<0.05）。本研究结果充分说明稻田各蜘蛛类群对不同生育期、不同品种水稻的褐飞虱均具有较强的捕食作用,是生物防治策略中的重要因素,应加强田间蜘蛛的保护工作和增强自然天敌的控害功能。     

不同致害性褐飞虱种群的DNA多态性研究

采用RAPD-PCR方法,对分别在水稻品种TN1、Mudgo和ASD7上纯化82代的3个不同致害性褐飞虱种群的基因组DNA多态性进行了分析。从205个RAPD引物中筛选出了具种群特异性的引物10个,结果表明： 尽管未达到统计意义上的显著差异,雌、雄两性试虫种群间的遗传多样性一般高于种群内,种群内的多样性则以未经抗性品种筛选的TN1种群最高。用相似性系数进行类平均距离法聚类,能将同一致害性种群的各个个体与其他致害种群分开,雌、雄虫均无一例外,表明不同致害性种群间存在明显的遗传分化;同时雌、雄两性试虫分别归为2组,提示褐飞虱致害性遗传可能与性染色体连锁。不同致害性种群存在特有条带,有可能用于进一步建立褐飞虱个体致害性检测的分子标记。     

Ranking common predators of Bemisia tabaci in Georgia (USA) agricultural landscapes with diagnostic PCR: implications of primer specific post-feeding detection time

Developing a successful biological control program relies on understanding predator–prey interactions in agroecosystem field settings. Among several methods used, molecular gut content analysis (MGCA) has become a popular method to measure predator contributions to pest control services. Once MGCA is applied to diagnose predator–prey interactions, the DNA detectability half-life is often applied to adjust for differences in prey digestion time among predators. Although MGCA best practices are well established, with many primers available, further work is needed to rank among published primers for MGCA. Using a combination of laboratory feeding trials and application of diagnostic MGCA to field collected predators, we investigated Bemisia tabaci post-feeding detection times in three dominant predator functional groups (chewing, piercing/sucking, and spiders). This was based on three published B. tabaci-specific primers. These data reveal that primer choice generated significantly different B. tabaci DNA half-lives in predator gut content. The primers with longer half-life resulted in higher field predation frequency estimation. Our field data using the primer with the longest half-life suggest several abundant predators, including Hippodamia convergens, Geocoris punctipes, Orius spp., Thomisidae spider, and fire ants (Solenopsis invicta), are actively feeding on B. tabaci in cotton fields. Orius spp. and fire ants were the most abundant predator species in our study area and contributed the most to B. tabaci control. Our results suggest that primers can be classified based on their specific DNA half-lives and can be used to address different ecological questions such as how to study time-specific predation detection (nocturnal or diurnal).

     

The influence of time and temperature on molecular gut content analysis： Adalia bipunctata fed with Rhopalosiphum padi

Gut content analysis is a useful tool when studying arthropod predator-prey interactions. We used polymerase chain reaction （PCR） technique to examine how detection of prey DNA in the gut content of predators was influenced by digestion time and temperature. Such knowledge is critical before applying PCR-based gut content analysis to field collected predators. Larvae of the two-spotted ladybeetle （Adalia bipunctata L.） were fed with the bird cherry-oat aphid （Rhopalosiphum padi L.） at either 21℃ or 14℃. After consuming one aphid, the predators were allowed to digest the prey for a range of time periods up to 24 hours. The influence of temperature on A. bipunctata feeding behavior was also recorded. From the fed larvae, total DNA was extracted and PCR reactions with R. padi specific primers were run. The number ofA. bipunctata that tested positive for R. padi DNA was negatively related to the length of digestion time. Temperature influenced larval feeding behavior but did not have a significant effect on R. padi DNA detection. After pooling the data from both temperature treatments we estimated the time point when R. padi DNA could be amplified from 50% of the fed A. bipunctata by PCR to be 4.87 hours. With such a rapid decrease in prey DNA detection success, positive PCR reactions will most likely be the result of predation events occurring shortly before capture. If a defined digestion temperature range has proven not to influence prey detection, PCR data obtained from predators collected within that particular range can be interpreted in the same way.     

Photosynthesis and translocation of assimilates in rice plants following phloem feeding by the planthopper Nilaparvata lugens (Homoptera: Delphacidae)

Experiments were conducted to measure the effect of feeding by the planthopper Nilaparvata lugens (St?l) on photosynthesis and the translocation of assimilates in rice plants, Oryza sativa L. We used mature japonica rice plants and applied the 13CO2 feeding method to evaluate those physiological effects. The photosynthetic rate was suppressed by N. lugens infestation, especially at the lower leaf position, with rates 30% lower than that of control plants at the booting stage. Leaf nitrogen concentration in infested plants was also lower than that in control plants. After flowering, the chlorophyll content and total plant dry weight were reduced by N. lugens. Stem and sheath dry weights were significantly reduced by N. lugens infestation, whereas panicle dry weight was not affected. Little effect was found on disruption in translocation of assimilates, even when 13CO2 was supplied to the infested leaves. Results suggested that removal of assimilates and reduction in photosynthesis by N. lugens have the greatest effect on growth and yield of rice plants as compared with the disruption in the translocation of assimilates. Plant death can occur by N. lugens infestation if the amount of energy supplied is less than that required for tissue maintenance.     

Laboratory and field assessments of prey-mediated effects of transgenic Bt rice on Ummeliata insecticeps (Araneida: Linyphiidae)

One major concern regarding the release of Bt rice is its potential impact through tritrophic interactions on nontarget arthropods, especially natural enemies. We studied the effects of two Bt transgenic rice varieties, TT9- 3 and KMD1, expressing Cry1Ab/Cry1Ac and Cry1Ab, respectively, on a predatory ground spider [Ummeliata insecticeps (B?senberg et Strand)] supplied with Bt rice-fed brown planthopper [Nilaparvata lugens (St?l)] nymphs. Although immunoassays confirmed that U. insecticeps ingested Bt insecticidal protein when supplied with Bt rice-fed N. lugens, no negative effects were found on its survival and development. Furthermore, the fecundity of U. insecticeps fed prey reared on Bt rice was not significantly different from that of those fed prey reared on non-Bt rice. A 3-yr field trial indicated that Bt rice did not significantly affect the population density of U. insecticeps in comparison with non-Bt rice. In conclusion, the Bt rice lines tested in this study had no adverse effects on the survival, developmental time, or fecundity of U. insecticeps in the laboratory or on population dynamics in the field.     

Cry1Ab杀虫蛋白在水稻-褐飞虱-拟水狼蛛食物链中转移与富集

采用ELISA方法检测了2个转cry1Ab基因水稻（Bt水稻）品系KMD1和KMD2不同生育期叶鞘内Cry1Ab杀虫蛋白的含量及其通过褐飞虱和拟水狼蛛的转移和富集情况。结果表明,这2个品系中抽穗期和黄熟期叶鞘内Cry1Ab的含量均显著低于苗期、分蘖期和孕穗期,KMD1和KMD2中Cry1Ab杀虫蛋白可以通过食物链转移到Bt水稻非靶标害虫褐飞虱及其天敌拟水狼蛛体内。褐飞虱在KMD1或KMD2上取食2 d后,体内均含有Cry1Ab杀虫蛋白,但连续取食2、4、6、8和10 d后,其体内含量并未因取食时间的延长而呈现明显增加的趋势。当拟水狼蛛捕食以KMD1或KMD2为食的褐飞虱时,在捕食2、4、6、8和10 d后,其体内均可检测到Cry1Ab杀虫蛋白,其含量并未随捕食时间的延长而明显上升,但均显著高于相应时间褐飞虱体内的含量。可见,该蛋白可通过水稻转移至褐飞虱,再转移至拟水狼蛛,并存在明显的富集现象,而这种富集并不随蜘蛛捕食时间的延长而加强。拟水狼蛛捕食以KMD1或KMD2为食的褐飞虱时,其捕食量未受到显著影响,其中肠酶粗提物对Cry1Ab杀虫蛋白具有明显的降解作用。     

一种褐飞虱病原真菌的分子生物学鉴定

从田间褐飞虱Nilaparvata lugens( St(a)l)罹病虫体分离得到一株绿僵菌,以该菌为供试菌体,采用氯化苄法、CTAB法以及裂解液法分别提取了该菌的基因组DNA;以ITS1和ITS4为绿僵菌通用引物,对供试菌株的rDNA ITS序列进行PCR扩增、琼脂糖凝胶电泳检测和序列分析,并在核酸序列数据库中进行同...     

Factors affecting detectability of prey DNA in the gut contents of invertebrate predators: a polymerase chain reaction-based method

This study aimed to determine factors that influence the detection of DNA of Plutella xylostella L. (Lepidoptera: Plutellidae) in the gut contents of arthropod predators when the polymerase chain reaction is used to amplify a diagnostic fragment of the gene coding for cytochrome oxidase subunit I. The effects of temperature, time since feeding, subsequent food intake, sex, weight, and species of predator on prey detectability were studied in the laboratory. Three types of predator were studied: the spider Venator spenceri Hogg. (Araneae: Lycosidae), a bug with sucking mouthparts, Nabis kinbergii (Reuter) (Heteroptera: Nabidae), and a coccinellid with chewing mouthparts, Hippodamia variegata (Goeze) (Coleoptera: Coccinellidae). In all experiments, the detectability of prey DNA was negatively correlated with time post‐feeding. The duration of detectability differed among the predator species. The time calculated for median detection success at 20 °C ranged from 49.6 h in V. spenceri to 36.1 h in N. kinbergii and 17.1 h in H. variegata. In H. variegata, but not in V. spenceri, the rate of detection decreased with increasing temperature. Subsequent food intake did not affect the detectability of DNA of P. xylostella in V. spenceri. In H. variegata, sex and weight of the predator did not influence detection of prey DNA. In addition, this study uncovered potential sources of error caused by detection of prey DNA following secondary cannibalistic and intraguild predation. The results provide essential information for the interpretation of prey detection data from field‐collected predators’ gut contents.     

褐飞虱Nl15基因的克隆及功能分析

【目的】植食性刺吸式口器昆虫的唾液蛋白参与调控植物抗虫防御反应,影响其对寄主植物的适应性。本研究旨在通过克隆褐飞虱Nilaparvata lugens重要唾液蛋白基因Nl15,调查其时空表达模式,明确其在褐飞虱致害性中的作用。【方法】基于褐飞虱IR56种群转录组数据,用RT-PCR克隆褐飞虱基因Nl15 cDNA序列,并进行生物信息学分析。利用qPCR检测其在褐飞虱TN1和IR56种群不同发育阶段(卵、1-5龄若虫和雌雄成虫)和雌成虫不同组织(头、胸、腹和足)中的表达模式。通过显微注射dsRNA对褐飞虱TN1和IR56种群的4龄若虫进行Nl15的RNAi,利用qPCR检测Nl15 RNAi后褐飞虱若虫中Nl15的相对表达量以及Nl15 RNAi后褐飞虱若虫取食3 d时水稻植株中防御相关基因(OsLecRK4, OsMPK10, OsWRKY24, OsLox, OsNPR1和OsGns5)的相对表达量,并生物测定Nl15 RNAi后褐飞虱的存活率以及成虫蜜露量和体质量增量。【结果】克隆了褐飞虱Nl15 cDNA序列(GenBank登录号：OK181113),其开放阅读框长1 008 bp;预测编码335个氨基酸,理论等电点为7.54,分子量为38.7 kD,含有23个氨基酸的信号肽序列和一个糖基化修饰位点,不存在跨膜结构域和其他已知的功能域;Nl15与灰飞虱Laodelphax striatellus同源蛋白氨基酸序列一致性为45%。发育表达谱结果表明,Nl15在褐飞虱各个发育阶段均表达,在3-4龄若虫中的表达量最高;组织表达谱结果表明,Nl15在褐飞虱雌成虫头部中的表达量最高,且在IR56种群头部中的表达量高于在TN1种群头部中的。RNAi实验结果表明,与注射dsGFP的对照组相比,注射dsNl15的处理组中Nl15的表达量显著降低了89.5%,褐飞虱的存活率以及成虫蜜露量和体质量增量均显著降低,上述6个水稻防御相关基因的表达量显著上调。【结论】褐飞虱IR56种群中的Nl15参与褐飞虱与水稻的防御与反防御分子互作。本研究为进一步阐述褐飞虱克服抗虫基因的机制及揭示昆虫与植物互作的分子网络提供了思路。