不同虫源和致害性的褐飞虱体内共生菌的种群动态

通过对不同地理种群和致害性的褐飞虱体内共生菌,在高温和抗性水稻品种作用下的种群动态研究表明,在正常湿度下褐飞虱体内共生菌的数量随若虫的生长发育而增加,5龄若虫期是共生菌增长的关键期;同一种群的褐飞虱秋季共生菌数量明显高于夏季。广西田间种群的短翅雌虫体内的共生菌数量显高于浙江省的杭州和龙游种群,经高温处理后,3个生物开共生菌数量均显减少,但其子代共生菌的数量在正常温度下马上恢复,与未经高温处理的褐飞虱共生菌数量基本相同,取食抗性品种的生物型1和田间种群的褐飞虱体内的共生菌数量均明显减少,取食抗性品种的第2代的共生菌又均比第1代少。这说明高温对共生菌的影响是暂的,而抗性品种的对共生菌的作用与褐飞虱种群对品种的适应性有关。     

共生菌在褐飞虱致害性变化中的作用

研究了不同虫源和致害性褐飞虱Nilaparvata lugens种群体内共生菌数量动态及其对褐飞虱在抗虫品种上的取食选择、生长发育、繁殖以及氨基酸转移酶活性的影响。结果表明,褐飞虱田间种群的致害性与其体内共生菌数量有关。广西南宁种群雌成虫体内的共生菌数量显著地高于浙江杭州和龙游两个虫源的雌成虫体内共生菌数量,而已纯化的3个不同致害性生物型体内的共生菌数量无显著差异。取食抗性品种能显著减少生物型Ⅰ雌成虫体内的共生菌数量。缺乏共生菌时,生物型Ⅰ、Ⅱ若虫对水稻品种TN1和ASD7的选择性增大,而对Mudgo的取食选择性下降。尽管缺共生菌的3个生物型在已适应的和不适应的感虫和抗虫品种上的若虫存活率和雌成虫产卵量均下降,若虫历期明显延长,但在已适应品种上的变化程度明显小于在不适应的抗虫品种上的变化程度。共生菌还明显影响成虫体内丙氨酸氨基转移酶和天门冬氨酸氨基转移酶的活性。这些结果证明体内共生菌的数量和质量在褐飞虱对水稻致害性的变化中发挥了重要作用。     

芽孢杆菌的引入对褐飞虱微生物群和生长发育的影响

【目的】明确褐飞虱Nilaparvata lugens体内可培养共生细菌的种类，探索其中芽孢杆菌类共生菌对褐飞虱微生物群和生长发育的影响。【方法】通过离体培养的方法，从褐飞虱两种不同致害性种群(TN1敏感种群和IR56高致害种群)中分离可培养共生细菌。通过16S rDNA测序对获得的可培养共生菌进行鉴定，在此基础上利用原位杂交研究了共生细菌在褐飞虱体内的分布。利用人工饲料添加抗生素和回补共生细菌的方式，研究了减菌和回补芽孢杆菌的处理对褐飞虱生长发育以及共生菌丰度的影响。比较了通过饲喂和显微注射对芽孢杆菌引入的影响，考察了芽孢杆菌的定殖与褐飞虱TN1种群致害性之间的相关性。【结果】利用离体培养法从褐飞虱中获得了15株不同的共生细菌，其中包括2株源于IR56高致害种群的芽孢杆菌类共生细菌BPH-S36和BPH-S33。原位杂交结果表明共生细菌在褐飞虱成虫唾液腺、肠道、脂肪体和雌虫内生殖器中均有分布，而在雄虫内生殖器中鲜有分布。体内共生细菌对褐飞虱的生长发育至关重要，共生细菌的减少导致褐飞虱存活率在第3和6天时显著下降，而回补共生芽孢杆菌BPH-S36或BPH-S33可使其存活率在第6天时显...     

褐飞虱不同致害性种群体内共生菌18S rDNA部分序列比较

分离纯化了褐飞虱3种不同致害性种群体内类酵母共生菌 (yeast-like symbionts, YLS),并对其18S rDNA基因序列进行了比较。结果表明,褐飞虱3种不同致害性种群体内类酵母共生菌18S rDNA均扩增出600 bp左右的片断。依据获得的18S rDNA特异性序列,结合已知真菌的18S rDNA部分序列,构建了不同宿主的YLS分子系统树。结果显示, 褐飞虱3种不同致害性种群体内的YLS同属子囊菌亚门（Ascomycotina）的核菌纲（Pyrenomycetes）,并与此纲中的Hypomyces chrysospermus亲缘关系相对最近。     

亚洲不同地理种群褐飞虱内共生菌Wolbachia的分子检测

褐飞虱Nilaparvata lugens Stl是水稻的重要害虫,而Wolbachia是其重要的次生共生细菌之一。为了解褐飞虱种群感染Wolbachia的情况,本研究对采自世界褐飞虱主要分布区的22个地理种群感染Wolbachia的情况进行了检测和分析。qPCR结果表明,除杭州2009年种群没有检测出Wolbachia外,其它21个种群中都有不同程度的Wolbachia感染。对Wolbachia的wsp基因序列的比较和进化分析表明,褐飞虱各地理种群Wolbachia都属于B群,并可进一步分为Ori和Con两个亚群。其中18个褐飞虱种群中的Wolbachia属于Ori亚群,广东清远和浙江桐乡褐飞虱种群中的Wolbachia则属于Con亚群,而菲律宾Iloilo褐飞虱种群分别检测到同时有2个亚群Wolbachia感染。本结果可为进一步研究褐飞虱与共生菌的共生关系、褐飞虱致害性变异和迁飞路线分析及利用Wolbachia防治褐飞虱提供依据。     

褐飞虱体内Arsenophonus、Wolbachia、Spiroplasma和Cardinium四种次生共生菌感染分析

【目的】Arsenophonus、Wolbachia、Spiroplasma和Cardinium是广泛分布于昆虫体内的次生共生菌,能对寄主昆虫的生殖和发育生理产生影响,明确不同地理种群褐飞虱体内4种共生菌的感染和系统发育关系可为利用共生菌防治害虫提供技术依据。【方法】利用Arsenophonus特异性23S rRNA基因引物,Wolbachia、Spiroplasma和Cardinium各自的16S rRNA基因引物PCR检测广西10个地市、海南海口及上海、江苏南京、湖南长沙共14个地理种群褐飞虱雌、雄虫体内共生菌感染情况,并对扩增到的基因片段进行测序、构建系统发育树。【结果】不同地区2014年和2015年褐飞虱体内Arsenophonus感染率最高的分别为河池种群(30%)和长沙种群(20%)、PWolbachia感染率最高为河池种群(20%)和南京种群(30%),而Spiroplasma和Cardinium在褐飞虱各种群内均未检测到。序列比对和发育分析表明,褐飞虱体内Arsenophonus与同翅目昆虫白蜡虫Ericerus pela Chavannes体内感染的该菌关系最近;而感染Wolbachia菌的系统发育分析表明,除海南种群有一个个体感染Wolbachia菌A群外,其余种群感染的全部为B群;同时在所有感染两类共生菌的个体中并未发现共感染现象。【结论】不同地理种群褐飞虱感染Arsenophonus和Wolbachia两类共生菌的感染率并不一致;总体上褐飞虱雌虫Wolbachia感染率要高于雄虫,褐飞虱体内感染的两类共生菌与其它昆虫体内的共生菌序列高度一致或类似,且在所有检测的地理种群中均未发现有Spiroplasma和Cardinium感染。     

稻飞虱酵母类胞内共生菌的组织学研究进展

褐飞虱Nilaparvata lugens体内酵母类共生菌在褐飞虱生长发育和繁殖中起着重要作用,并影响着褐飞虱对寄主植物的致害性变异。该文系统总结了国内外已有的研究成果,讨论了褐飞虱共生菌的分类地位、形态特征、在寄主发育中的功能、侵染途径等,提出了开展褐飞虱生物型形成过程中体内共生菌变异的研究建议,以便利用共生菌来监测田间不同生物型褐飞虱的发生情况,最终实现控菌防虫和对不同生物型的准确预测。     

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。     

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。     

我国褐飞虱若干地理种群致害性的研究

褐飞虱致害性变异是水稻品种抗性利用的一个重要障碍,监测田间种群的致害性对水稻抗虫品种的培育和利用有重要意义。本文采用蜜露量法对采集自云南勐海、贵州遵义和旧州、广西南宁、湖南衡阳以及浙江富阳等6个褐飞虱田间种群对IR26(含Bph1)和IR42(含bph2)的致害性进行了研究,发现勐海种群对IR26、IR42均有较强的致害性,遵义、旧州、南宁和衡阳种群仅对IR26致害性较强,而对IR42相对较弱;富阳种群对IR26、IR42的致害能力均相对较弱。采用SSST法进一步比较了西南稻区云南勐海、贵州遵义和旧州褐飞虱种群对TN1、IR26、Mudgo、ASD7、IR36、IR42、IR56、Rathu Heenati、Ptb33和Babawee等10个水稻品种的致害性,结果表明:勐海种群的致害性最强,主要体现在对Rathu Heenati、IR56、Ptb33等含Bph3基因的致害性显著强于旧州种群和遵义种群;后两者间,除旧州种群对IR36致害性显著较强外,对其他品种的致害性均无显著差异。总体上,处于东南季风带之外、云南西南部的勐海褐飞虱种群致害性明显强于来源于东南季风带之内的5个褐飞虱种群,进一步为我国西南稻区西部与云贵高原东缘及以东地区分属不同的褐飞虱迁飞场提供了重要依据。     

不同地理种群褐飞虱的抗逆性

研究了不同地理种群褐飞虱对水稻抗性品种、若虫高密度、高温、饥饿和杀虫剂等逆境条件的抗性。结果表明,广西和云南种群褐飞虱对抗虫品种的适应性比浙江种群强而广,浙江种群只能适应抗虫品种ＩＲ２６而广西和云南种群则还能适应抗性更高的ＡＳＤ７和Ｒａｔｈｕ Ｈｅｅｎａｔｉ。广西和云南种群较能适应高密度环境,而浙江种群的适应性较差,其雌虫短翅率与若虫密度呈显著负相关。虽然广西和云南种群对高温的忍受性比浙江种群的强     

Effects of elevated CO2 and transgenic Bt rice on yeast‐like endosymbiote and its host brown planthopper

The effects of elevated CO₂ (750 vs. 375μl/l) on population abundances and metabolism enzyme of AChE and protective enzymes of SOD, POD and CAT in brown planthoppers (BPH) Nilaparvata lugens, and on size and abundances of yeast‐like endosymbiotes (YLES) were studied as BPH fed Bacillus thuringiensis (Bt) rice expressing pure Cry1Ab after successively two generations in open‐top chambers. The results indicated that: (1) Brachypterous and macropterous subpopulations and total population increased with elevated CO₂. Significant increases were found as BPH fed non‐transgenic rice while only significant increase as macropterous‐BPH fed Bt rice. (2) The responses of brachypterous and macropterous‐BPH to Bt rice were different. Brachypterous‐subpopulation significantly decreased (13.6%) while macropterous ones significantly increased (43.8%) as fed Bt rice relative to non‐transgenic rice at elevated CO₂. (3) Elevated CO₂ only significantly inhibited AChE activity as brachypterous‐BPH fed non‐transgenic rice. Significant increases in POD and SOD, and significant decrease in CAT were found as brachypterous‐BPH fed Bt rice, while significant increases in CAT and significant decrease in POD were also observed as fed non‐transgenic rice in elevated CO₂ relative to ambient CO₂. (4) Bt rice significantly inhibited POD and SOD activity at ambient CO₂, while only significantly enhanced SOD activity at elevated CO₂. (5) Elevated CO₂ significantly decreased YLES per mg/head of brachypterous‐BPH females while only significantly decreased YLES per mg/head as brachypterous‐BPH males fed Bt rice. And there were significant differences in YLES width or length between females and males. Elevated CO₂ can markedly affect the symbiosis relationship between YLES and BPH through the bottom‐up forcing on BPH physiological metabolism. And the damage inflicted by BPH on rice, irrespective of the presence of insecticidal genes, is predicted to be higher at elevated CO₂. Furthermore, transgenic Bt rice can also exacerbate emigrating‐macropterous‐BPH occurring especially at elevated CO₂.     

THE VIRULENCE CHANGE AND DAMAGE CHARACTERISTICS OF VARIOUS GEOGRAPHIC POPULATIONS OF BROWN PLANTHOPPER*

Abstract The virulence change and damage characteristics of brown planthopper (BPH), Nilaparvata lugens Stal, populations collected from Guangxi, Yunnan and Zhejiang Provinces were studied by using various resistant rice varieties. The results indicated that the Zhejiang population could successfully infest varieties IR26 and Mudgo, showing the characters of BPH biotype 2 after 1989. The Guangxi population, however, could adapt the resistant varieties IR26, Mudgo and ASD7 after 1991. In 1997, the nymphal survival indices of Guangxi and Yunnan populations on Rathu Heenati (Bph3) were up to 70. 9 and 66. 7 respectively. This may imply that the BPH populations in Guangxi and Yunnan have shifted into a new biotype. In free-choice test, there was obvious difference in the feeding preference of nymphs and adults of Zhejiang population, the adult preferred to feeding on resistant variety IR26 and the nymphs chose the hosts randomly. Under 26 C and L12: D12, the nymphal duration of Zhejiang population was the longest in the three tested populations. For Guangxi population, the nymphal duration on ASD7 was significantly longer than on TN1, although it could infest resistant variety ASD7. The nymphal survival rate of Zhejiang population fed on IR26 was higher than those fed on TN1 and ASD7. On varieties IR26 and ASD7, an obviously higher nymphal survival rate was observed in Yunnan population as compared to those of Zhejiang and Guangxi populations. These results indicate that difference in virulence and damage characteristics could be found in different geographical populations of BPH.     

不同温度下浸水对褐飞虱生存、繁殖及其寄主的影响

不同温度下浸水对褐飞虱卵孵化、若虫存活率、成虫取食和繁殖及其寄主水稻的影响研究结果表明 ,在相对低温2 2℃和 2 1～ 2 6℃的自然变温条件下 ,产卵后立即浸水对浙江和广西 2个褐飞虱种群的卵孵化影响均不明显 ,而在产卵后4 d浸水对浙江种群的卵孵化有显著的抑制作用。 2 8℃浸水 5d和 35℃浸水 1 d后卵孵化率显著下降且孵化进程延长。卵浸水后的初孵若虫的存活率随卵浸水时间的延长明显下降。稻株浸水对其体内所含的昆虫必需氨基酸影响较大 ,其总量减少 ,而稻株内可溶性糖含量却显著增加 ,导致褐飞虱在浸水稻株上的若虫生存率、成虫取食量和繁殖力均明显下降     

褐飞虱的迁人和翅型分化规律的研究

褐飞虱Nilaparvata lugens(Stal)属迁飞性水稻害虫,其成虫有短翅和长翅两种翅型。 褐飞虱迁入雌虫均未经过交配,其短翅型由一对显性等位基因控制。翅型分化同时受到遗传和内分泌系统的协调控制,外界条件如密度和寄主等因子通过内分泌系统来影响翅型的分化。褐飞虱翅型分化的敏感龄期雌虫为1～3龄,而雄虫为1-5龄;不同若虫蜜度处理对褐飞虱成虫的前翅形成有一定的影响。分蘖期水稻饲养的褐飞虱短翅型比例明显高于孕穗期水稻饲养的褐飞虱。不同密度下各生物型间的翅型分化差异不显著。迁入地的浙江各种群属温带型,其雌虫短翅率低且与密度呈显著负相关;菲律宾热带种群雌虫在不同密度下均为短翅型,而雄虫的短翅率随密度增加而上升。广西种群接近热带型,其雌虫短翅率高但不随密度而变化。试验各种群的雄虫在中等密度甚至高密度时其短翅率出现最高。     

中国与越南褐飞虱和白背飞虱生物型研究

【目的】监测我国与越南褐飞虱Nilaparvata lugens St?l和白背飞虱Sogatella furcifera Horvath生物型,为抗虫育种工作提供指导。【方法】应用群体集团检测法和蜜露量检测法研究了中国广西、云南、河南、湖南、重庆、贵州和越南河内、河静、顺化、胡志明市和九龙江田间褐飞虱和白背飞虱的致害特性和生物型组成结构。【结果】我国主要稻区(除云南思茅外)和越南中北部的田间褐飞虱以Ⅱ型的比例多,对含Bph1、bph2基因的鉴别品种表现为致害;云南思茅的田间褐飞虱以Ⅱ+Ⅱ型的比例多,对含Bph1、bph2和bph4基因的鉴别品种表现为致害或强致害;越南胡志明市、九龙江的田间褐飞虱以Ⅱ+Ⅱ型的比例多,对含Bph1、bph2、Bph3、bph4基因的鉴别品种主要表现为致害或强致害。我国白背飞虱以Ⅰ型比例较多;越南顺化和河内以Ⅱ型比例多;所有监测点白背飞虱的致害特性总体表现为对含Wph1、Wbph2基因的鉴别品种的致害能力较强,对Wbph3的致害能力表现不一,对含Wph5基因的鉴别品种表现为中等致害。【结论】抗虫育种选择抗源时,不要选含Bph1、bph2基因的水稻品种作为褐飞虱抗源,不要选含基因Wbph1或Wbph2的水稻品种作为白背飞虱抗源。     

Rice planting systems, global warming and outbreaks of Nilaparvata lugens (Stål)

Brown Planthopper (BPH, Nilaparvata lugens (St?l)) is one of the most serious pests of rice in both temperate and tropical regions of East and South Asia and has become especially problematic over the past few years. In order to analyze the effect of the change of rice cropping system on the population dynamics of BPH, field surveys of the occurrence and distribution of BPH were performed and other relevant data, including light trap data and ovary dissection data were collected in nearly 40 Chinese counties encompassing six provinces (or municipalities), including Hainan, Guangxi, Anhui, Shanghai, Fujian and Guangdong from April to October in 2007.The mixed planting areas of single- and double-cropping rice in China include Hubei, South and Central Anhui, North Hunan, and North Jiangxi. In these areas, double-cropping rice has now been greatly reduced and single-cropping rice has been rapidly increasing since 1997. The surveys revealed that when the immigration peak of BPH occurred in June and July, the single-cropping rice was at the tillering to booting stage and fit for BPH, but early rice had already matured and most of late rice had not yet been transplanted. BPH immigrants from southern rice areas prefer to inhabit and breed in single-cropping rice paddies. Moreover, farming activities between early rice and late rice interrupted the continuous growth of BPH populations in double-cropping rice paddies. As a result, in comparison with data collected 30 years ago, the spatiotemporal dynamics and migration patterns of BPH have dramatically changed in the lower-middle reaches of the Yangtze River. In the mixed planting areas, due to their high suitability, the BPH population in single-cropping rice grew so quickly that it caused serious local damage and there was mass emigration of macropterous progeny to the Yangtze River Delta in late August and early September.Global warming may also affect BPH populations, where results suggest steadily warmer autumns have occurred from the 1990s on, with such conditions gradually the norm. The combination of 'cooler summer' and 'warmer autumn' are conditions known to promote outbreaks of BPH in the lower-middle reaches of the Yangtze River. Immigrant BPH arrivals in late August and September now cause serious damage to late-maturing mid-season rice and late rice in the lower-middle reaches of the Yangtze River.