中国部分地区烟粉虱生物型种类及系统发育关系分析

烟粉虱是一种危害严重的世界性害虫,是一个快速进化的复合种.本文利用mtCOI分子标记方法,对2010和2011年采自我国9个省(市)的33个烟粉虱种群进行了生物型鉴定和系统发育分析.结果表明： 我国目前存在着B型、Q型、ZHJ-1型、ZHJ-3型、An型以及Nauru型等6种生物型,且不同生物型的分布是不均匀的.遗传距离及系统发育树的分析结果显示,海南省的An型和台湾省的An型聚为一支,为同一来源;中国B型与来自法国和乌干达的B型的亲缘关系较近,同源性达到99%以上;中国的Q型与来自摩洛哥和法国的Q型聚为一个分支,而来自以色列和土耳其的Q型烟粉虱单独聚为一支,说明中国的Q型烟粉虱与来自地中海西部的Q型烟粉虱亲缘关系更近,可以推断中国的Q型烟粉虱的起源地为地中海西部地区.
     

江苏地区烟粉虱发生危害及其生物型监测

【目的】随着Q型和B型烟粉虱Bemisia tabaci(Gennadius)在全世界范围内的传播和入侵,两种入侵烟粉虱的发生危害和种群扩张越来越受到人们的关注。本文对2010—2013年江苏省内烟粉虱的发生危害程度及其生物型的分布进行了全面的调查研究。【方法】应用室内镜检鉴定烟粉虱的发生危害程度,同时结合mt COⅠ分子标记快速检测法鉴定烟粉虱的生物型。【结果】从时间来看,烟粉虱的发生危害程度在2011年最严重,而在2013年最低,呈"先上升后下降"的趋势。从不同地区来看,苏南和苏北地区呈现出"先上升后下降"的趋势,而苏中地区与之相反。从寄主来看,烟粉虱在茄子,黄瓜和南瓜上的发生危害程度最严重。【结论】烟粉虱在江苏省内的发生危害程度因时间,地区和寄主的不同而差异显著,这种差异与烟粉虱不同生物型之间的竞争取代没有直接联系。虽然Q型烟粉虱在江苏省内占据了优势地位,但是B型烟粉虱在苏南和苏北地区仍然占据了一定的比例,并且有逐步上升的趋势。     

新疆地区烟粉虱生物型的区域分布及其携带的番茄黄化曲叶病毒检测

【目的】明确不同烟粉虱Bemisia tabaci (Gennadius)生物型在新疆地区的区域分布及其携带番茄黄化曲叶病毒的情况。【方法】本文利用烟粉虱mtDNA COⅠ基因片段作标记以及番茄黄化曲叶病毒(tomato yellow leaf curl virus, TYLCV)特异性引物, 鉴定新疆不同地区烟粉虱生物型, 并检测其携带TYLCV情况。【结果】石河子市、 博乐市、 岳普湖县、 泽普县、 叶城县、 莎车县、 墨玉县、 和田市与和田县烟粉虱属于B型烟粉虱, 且720 bp COⅠ基因序列一致性为100%; 阿克苏市、 哈密市、 鄯善县、 乌鲁木齐市、 克拉玛依市与伊宁市烟粉虱属于Q型烟粉虱, 720 bp COⅠ基因序列一致性为99.9%~100%; 吐鲁番市与库尔勒市兼有B型烟粉虱和Q型烟粉虱。伊宁市、 乌鲁木齐市、 鄯善县、 哈密市、 阿克苏市、 库尔勒市、 莎车县、 和田县和墨玉县的烟粉虱样本携带TYLCV, 除2011年底已经暴发危害的莎车县外, 其他区域均为番茄黄化曲叶病毒病发生的高风险区域。【结论】本研究为新疆地区烟粉虱及番茄黄化曲叶病毒病的科学防控提供依据。     

西藏发现Q型烟粉虱

【目的】调查西藏自治区烟粉虱Bemisia tabaci(Gennadius)的发生情况。【方法】从西藏拉萨采集到烟粉虱各个虫态,采用3D数码显微镜观察所采集烟粉虱的形态特征,利用mt COⅠ分子标记检测烟粉虱的生物型。【结果】明确并详细描述了烟粉虱各形态特征,mt COⅠ分子标记检测显示西藏采集到的烟粉虱为Q生物型。【结论】在形态学鉴定的基础上,分子生物学鉴定该粉虱为Q型烟粉虱,这是Q型烟粉虱在西藏自治区发生的首次报道。     

一种含特异性侵染内共生菌Rickettisa massilia的烟粉虱种群建立方法

【目的】烟粉虱Bemisia tabaci(Gennadaius)广泛分布于全球除南极洲外各大洲的90多个国家和地区,其寄主范围广,危害严重,属于农业上的重要害虫。烟粉虱体内的内共生菌有助于其扩散传播,提高其寄主适合度,但其体内的内共生菌大都为次生内共生菌,很难进行体外培养,因此,加大了其内共生菌研究的难度。【方法】采集我国部分地区的烟粉虱种群,然后利用基于mt DNA COⅠ基因的种特异性引物进行种群生物型鉴定,再用内共生菌的特异性引物来鉴定含有特异性侵染内共生菌种群,确定个体含内生菌类型后,采取继代饲养方法建立种群。【结果】经鉴定烟粉虱的生物型为Q型(MED隐种),经过5代种群筛选后,可以建立稳定遗传的含特异性侵染内共生菌Rickettisa massilia菌的烟粉虱种群。【结论】通过本方法可以获得稳定的含有特异性侵染内共生菌Rickettisa massilia菌的烟粉虱种群。该方法对研究烟粉虱与体内内共生菌、植物三者之间以及烟粉虱体内某几种内共生菌之间的互作有重要意义,同时也为明确烟粉虱体内Rickettisa菌的作用研究打下基础。     

我国烟粉虱自然种群中存在广泛的Wolbachia感染现象

Wolbachia是广泛分布于节肢动物生殖组织中的细胞质遗传的一类共生菌,可以引起宿主生殖行为的改变.烟粉虱是重要的农业害虫,已有的研究表明烟粉虱中存在Wolbachia的感染,但所检测到的感染率不高,并且迄今在烟粉虱中所检出的Wolbachia均属于B组群.该研究从我国河北、新疆、北京、山东、浙江、广西、海南、广州和福建采集到18个烟粉虱地理种群,首先基于ITS1 rDNA克隆测序鉴定了烟粉虱的生物型,然后采用自行设计的Wolbachia 16S rDNA及wsp基因的专用引物对所采集到的烟粉虱种群进行了分子检测.结果表明几乎所有的烟粉虱自然种群中都存在Wolbachia感染,同时还发现B/Q生物型烟粉虱中主要感染A组群Wolbachia,而非B/Q生物型烟粉虱中存在大量的超感染现象.该研究显示烟粉虱自然种群中Wolbachia的感染率比预想的要高得多,而分子检测方法的灵敏度可能是影响Wolbachia感染率研究的关键因素之一.     

烟粉虱B型和Q型群体遗传结构的RAPD分析

近20年来,烟粉虱B型传入世界各地并暴发成灾,成为一种重要的农业入侵害虫; 烟粉虱Q型则是近几年引起人们高度重视的一种新的入侵生物型,目前已传入许多国家并造成一定危害。本文利用RAPD分子标记对烟粉虱B型和Q型不同地理种群的遗传结构进行了分析。结果表明：（1）引物H16对烟粉虱B型不同种群扩增的特异带,能有效区分烟粉虱B型和Q型、浙江非B/Q型种群;（2）烟粉虱Q型种群各项遗传多样性指数均比烟粉虱B型的要高;（3）我国烟粉虱Q型来自伊比利亚半岛的可能性比来自中东地区的可能性要大。另外,聚类分析结果提示,RAPD分子标记能有效地区分烟粉虱不同生物型,但可能不适用于生物型之间亲缘关系分析。     

一种含特异性侵染内共生菌Rickettisa massilia的烟粉虱种群建立方法

【目的】烟粉虱Bemisia tabaci(Gennadaius)广泛分布于全球除南极洲外各大洲的90多个国家和地区,其寄主范围广,危害严重,属于农业上的重要害虫。烟粉虱体内的内共生菌有助于其扩散传播,提高其寄主适合度,但其体内的内共生菌大都为次生内共生菌,很难进行体外培养,因此,加大了其内共生菌研究的难度。【方法】采集我国部分地区的烟粉虱种群,然后利用基于mt DNA COⅠ基因的种特异性引物进行种群生物型鉴定,再用内共生菌的特异性引物来鉴定含有特异性侵染内共生菌种群,确定个体含内生菌类型后,采取继代饲养方法建立种群。【结果】经鉴定烟粉虱的生物型为Q型(MED隐种),经过5代种群筛选后,可以建立稳定遗传的含特异性侵染内共生菌Rickettisa massilia菌的烟粉虱种群。【结论】通过本方法可以获得稳定的含有特异性侵染内共生菌Rickettisa massilia菌的烟粉虱种群。该方法对研究烟粉虱与体内内共生菌、植物三者之间以及烟粉虱体内某几种内共生菌之间的互作有重要意义,同时也为明确烟粉虱体内Rickettisa菌的作用研究打下基础。     

湖北省烟粉虱生物型鉴定

本研究利用mtDNA COI基因片段作标记,通过序列分析对湖北省烟粉虱Bemisa tabaci(Gennadius)生物型进行了鉴定。结果表明当阳、仙桃、天门、监利、宜昌和武汉6个地理种群烟粉虱的720bp COI基因序列的同源性高达100%,且该6个地理种群烟粉虱与USA-Q型烟粉虱、湖北Q型烟粉虱的同源性也高达100%。枣阳地区烟粉虱与Israel-B型烟粉虱、我国浙江B型烟粉虱、我国湖北B型烟粉虱的同源性为100%。由此可见当阳、仙桃、天门、监利、宜昌和武汉6个地区烟粉虱生物型属于Q型,仅枣阳地区烟粉虱属于B型。     

烟粉虱传毒相关蛋白基因的克隆及其表达量分析

【目的】烟粉虱Bemisia tabaci(Gennadius)是一种世界性的入侵性害虫,其传播的番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)给我国番茄生产造成巨大经济损失。为了阐明烟粉虱传播双生病毒的机理,本文拟研究明确B和Q烟粉虱体内传毒相关蛋白Gro EL基因及其表达量。【方法】采用特异性引物克隆了B和Q烟粉虱内共生菌编码的传毒相关蛋白Gro EL基因,并进行序列分析;并利用荧光定量PCR技术检测两种生物型及其获取双生病毒前后该基因的相对表达量。【结果】烟粉虱内共生菌Hamiltonella编码的Gro EL基因全长为1 668 bp,编码555个氨基酸;B、Q烟粉虱该基因的核苷酸序列相似性为99.94%,氨基酸同源性为99.82%;带毒烟粉虱中Gro EL基因的表达量显著高于未带毒的对应生物型烟粉虱;无论带毒与否,Q烟粉虱该基因的表达量均显著高于B(P<0.05)。【结论】烟粉虱携带TYLCV后可诱导Gro EL的表达量升高,B和Q烟粉虱中Gro EL基因及其表达量均存在差异,这可能是B和Q烟粉虱传毒效率存在显著差异的原因之一。     

B型烟粉虱与浙江非B型烟粉虱的竞争

为了了解近年来入侵中国的B型烟粉虱(Bemisiatabaci)取代本地非B型烟粉虱的潜能,在室内将一个B型与一个浙江非B型烟粉虱种群混合饲养在不同寄主植物上,跟踪观察混合种群中两个生物型个体数量相对比例的变化。结果表明,当两种生物型在棉花(Gossypiumhirsutum)上以相同初始数量共存竞争时,经过6代,非B型完全被B型替代;而在西葫芦(Cucurbitapepo)上以相同初始数量共存竞争时,只经过2代,非B型即完全被B型替代。在棉花上,即使以非B型占87%、B型占13%开始共存竞争,经过225d后,非B型也完全被B型替代。这说明B型烟粉虱具有在短期内竞争取代浙江非B型烟粉虱的能力。经分析,B型除了寄主范围比非B型的宽这一点对其竞争有利外,较强的内在竞争潜能也是其能成功入侵并替代本地非B型烟粉虱的一个重要原因。     

Further spread of and domination by Bemisia tabaci (Hemiptera: Aleyrodidae) biotype Q on field crops in China

The sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), causes severe crop losses to many crops. The worst of these losses are often associated with the invasion and establishment of biotypes B and Q of this pest. Previous research in 2007 showed that biotype Q occurred with other biotypes in most field populations in China. To determine the current status of the biotype composition in the field, an extensive survey covering mainly eastern parts of China was conducted in 2009. Using polymerase chain reaction primers specific for the mitochondrial cytochrome oxidase I of biotypes B and Q and gene sequencing, we determined the biotypes composition in 61 whitefly populations and their distribution across 19 provinces in China. Our research revealed that only biotypes B and Q have been found in the field in 2009 in China. Among them, biotype Q was dominant in 44 locations (100.0%) and biotype B was dominant in 17 locations (100.0%). The current survey indicates that biotype Q has rapidly displaced biotype B in most locations in China.     

Comparison of performance on different host plants between the B biotype and a non-B biotype of Bemisia tabaci from Zhejiang, China

The capacity of the B biotype of the whitefly, Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae), to invade has often been linked to its presumably wider host range than the non‐B indigenous biotypes. However, there are few experimental studies of the relative performance of the B biotype and non‐B biotypes on different host‐plant species. Here, we compared the performance of the B biotype and an indigenous non‐B biotype (China‐ZHJ‐1) of B. tabaci from Zhejiang, China on five commonly cultivated host plants, each from a different family: cotton, tobacco, cabbage, squash, and kidney bean. We also examined the effect of rearing host plants on the performance of the B biotype. Overall, the performance of the B biotype on the five species of plants was much better than that of the indigenous non‐B population. On tobacco, cabbage, and kidney bean, no individuals of ZHJ‐1 completed development to adulthood, whereas the B biotype developed successfully from egg to adult on all three plants. On squash, the B biotype survived better, developed to adulthood earlier and had a higher fecundity than ZHJ‐1. The two biotypes performed more equally on cotton, but even on this plant the B biotype female adults lived nearly twice as long as that of ZHJ‐1 and may have realized a higher life‐time fecundity. The B biotype also showed a substantial capacity to acclimatize to alternative host plants for improved survival and reproduction, on both highly suitable and marginally suitable host plants. We conclude that the host range of the B biotype of B. tabaci may be much wider than those of some indigenous biotypes, and this advantage of the B biotype over the non‐B biotypes may assist in its invasion and displacement of some indigenous biotypes in the field.     

Identification of Aphis gossypii Glover (Hemiptera: Aphididae) Biotypes from Different Host Plants in North China

Background

The cotton-melon aphid, Aphis gossypii Glover (Hemiptera: Aphididae), is a polyphagous species with a worldwide distribution and a variety of biotypes. North China is a traditional agricultural area with abundant winter and summer hosts of A. gossypii. While the life cycles of A. gossypii on different plants have been well studied, those of the biotypes of North China are still unclear.

Results

Host transfer experiments showed that A. gossypii from North China has two host-specialized biotypes: cotton and cucumber. Based on complete mitochondrial sequences, we identified a molecular marker with five single-nucleotide polymorphisms to distinguish the biotypes. Using this marker, a large-scale study of biotypes on primary winter and summer hosts was conducted. All A. gossypii collected from three primary hosts—hibiscus, pomegranate, and Chinese prickly ash—were cotton biotypes, with more cotton-melon aphids found on hibiscus than the other two species. In May, alate cotton and cucumber biotypes coexisted on cotton and cucumber seedlings, but each preferred its natal host. Both biotypes existed on zucchini, although the cucumber biotype was more numerous. Aphids on muskmelon were all cucumber biotypes, whereas most aphids on kidney bean were cotton biotypes. Aphids on seedlings of potato and cowpea belong to other species. In August, aphids on cotton and cucumber were the respective biotypes, with zucchini still hosting both biotypes as before. Thus, the biotypes had different fitnesses on different host plants.

Conclusions

Two host-specialized biotypes (cotton and cucumber) are present in North China. Hibiscus, pomegranate, and Chinese prickly ash can serve as winter hosts for the cotton biotype but not the cucumber biotype in North China. The fitnesses of the two host-specialized biotypes differ on various summer hosts. When alate aphids migrate to summer hosts, they cannot accurately land on the corresponding plant.     

Analysis of genetic diversity among different geographical populations and determination of biotypes of Bemisia tabaci in China

Abstract:  Analysis of the genetic diversity among 27 different geographical populations of Bemisia tabaci and determination of biotypes of B. tabaci in China based on amplified fragment-length polymorphism (AFLP) and the mitochondrial cytochrome oxidase I (mtDNA COI) gene sequences were conducted. In AFLP assay, the use of five primer combinations selected from 64 primer combinations allowed the identification of 229 polymorphic bands (97.03%) from 60 to 500 bp, suggesting abundant genetic diversity among different geographical populations of B. tabaci. To further identify biotypes of B. tabaci in China, the mtDNA COI gene sequences of nine representative populations from China, Israel and Spain were obtained. Molecular phylogenetic tree based on AFLP and mtDNA COI gene analyses revealed the presence, in China, of at least four different genetic groups of B. tabaci. B biotype, Q biotype and two non-B/Q biotype. B biotype was distributed nationwide. Q biotype was present only in the local region of China including the YunNan province and BeiJing city. This was also the first report about the invasion of Q biotype into China. Of the other two non-B/Q biotype groups, one was found in ShanDong and HeBei provinces, and another in ZheJiang province. The non-B/Q biotype ZheJiang population showed very high similarity with another Asian population India-IW ( AF110704 ) in mtDNA COI sequences and was possibly a Chinese indigenous population. The close monitoring of the Q biotype in locales of China where commercial plants were exported or imported, is now essential to avoid the further accidental distribution of the Q biotype.     

华东、华南和西南地区烟粉虱的种群及寄主分化

烟粉虱Bemisia tabaci是一个复合种,它具有的生物型分化、较强的传播病毒的能力和抗药性、较快的繁殖速率等特征使其成为我国农业生产中重要害虫之一。本研究利用细胞色素线粒体氧化酶Ⅰ基因,对采集自江苏、广东和云南三省的烟粉虱样本进行了生物型鉴定,并对烟粉虱生物型与寄主植物之间的关联性开展了调查。结果表明,在广东和云南省,都存在未鉴定的土著种群与入侵的B型、Q型共存的现象;同时,在本研究中广东省尚未采集到Q型烟粉虱,而在江苏采集到的粉虱样本全部为入侵型。研究结果还表明,相对于入侵种而言,土著种群显示出更强的寄主植物趋同性;丰富的寄主植物以及本身具有的多食性特性有助于B型、Q型等生物型在世界各地的广泛入侵。     

烟粉虱生物型的监测及其遗传结构研究

烟粉虱Bemisia tabaci(Gennadius)是一种重要的农业害虫并包括许多生物型,其中B型和Q型是入侵性较强的2种生物型。文章着重介绍近年来在烟粉虱生物型的监测及其遗传结构方面的研究进展。B型烟粉虱和Q型烟粉虱这2个生物型均已入侵我国,其中多数地区烟粉虱是B型烟粉虱,局部地区有Q型烟粉虱并呈现不断蔓延趋势。微卫星(SSR)分子标记分析结果表明我国B型烟粉虱的入侵来源具有多元化,而云南地区Q型烟粉虱来源比较单一。化学农药的使用能够影响室内种群的遗传结构,降低种群的遗传多样性。基于RAPD、ISSR分子标记的分析结果表明,Q型烟粉虱种群各项遗传多样性指数均比B型烟粉虱的高。今后加强烟粉虱入侵生物型的遗传结构及其种群动态关系等方面的研究,对于揭示烟粉虱的灾变机制及其控制具有重要的意义。     

Rapid spread of tomato yellow leaf curl virus in China is aided differentially by two invasive whiteflies

Background

Tomato yellow leaf curl virus (TYLCV) was introduced into China in 2006, approximately 10 years after the introduction of an invasive whitefly, Bemisia tabaci (Genn.) B biotype. Even so the distribution and prevalence of TYLCV remained limited, and the economic damage was minimal. Following the introduction of Q biotype into China in 2003, the prevalence and spread of TYLCV started to accelerate. This has lead to the hypothesis that the two biotypes might not be equally competent vectors of TYLCV.

Methodology/Principal Findings

The infection frequency of TYLCV in the field-collected B. tabaci populations was investigated, the acquisition and transmission capability of TYLCV by B and Q biotypes were compared under the laboratory conditions. Analysis of B. tabaci populations from 55 field sites revealed the existence of 12 B and 43 Q biotypes across 18 provinces in China. The acquisition and transmission experiments showed that both B and Q biotypes can acquire and transmit the virus, however, Q biotype demonstrated superior acquisition and transmission capability than its B counterparts. Specifically, Q biotype acquired significantly more viral DNA than the B biotype, and reached the maximum viral load in a substantially shorter period of time. Although TYLCV was shown to be transmitted horizontally by both biotypes, Q biotype exhibited significantly higher viral transmission frequency than B biotype. Vertical transmission result, on the other hand, indicated that TYLCV DNA can be detected in eggs and nymphs, but not in pupae and adults of the first generation progeny.

Conclusions/Significance

These combined results suggested that the epidemiology of TYLCV was aided differentially by the two invasive whiteflies (B and Q biotypes) through horizontal but not vertical transmission of the virus. This is consistent with the concomitant eruption of TYLCV in tomato fields following the recent rapid invasion of Q biotype whitefly in China.     

Biotype status and distribution of Bemisia tabaci (Hemiptera: Aleyrodidae) in Shandong province of China based on mitochondrial DNA markers

Bemisia tabaci has caused significant crop losses in China during the last decade. Recent research has shown that two potentially invasive variants, biotypes B and Q, have been found in several regions of China. Our objective was to determine the biotype status and the distribution of B. tabaci in Shandong province, an important agricultural region of China. Based on mitochondrial DNA markers, both biotypes B and Q were detected, with B being the predominant biotype. The results indicate that the more recently introduced biotype Q has not only been located in China but also has established and spread in some regions.     

Preference and performance responses of California grape phylloxera to different Vitis rootstocks

Laboratory experiments on host preference and performance were simultaneously conducted with Cabernet Sauvignon, AXR#1, and 5C Vitis rootstocks for the California biotypes A and B grape phylloxera. Preference bioassays recorded over a 3-day period indicated that phylloxera exhibit differential host choice. AXR#1 is antixenotic to biotype A, and 5C is antixenotic to both phylloxera biotypes. Preferences observed with biotype B for Cabernet Sauvignon and AXR#1 were not statistically different, whereas biotype A had shown a significant preference for Cabernet Sauvignon. On 5C the preferences scored were significantly lower than on either of the other two rootstocks, and there was no significant difference beetween biotypes. Performance bioassays recorded over a 29-day period indicated that survival, development, and reproduction of both biotypes were affected by Vitis rootstock. Both biotypes were unable to survive or develop on 5C suggesting the presence of antibiotic resistance in 5C against these phylloxera biotypes. The antibiotic effects observed with AXR#1 against biotype A were expressed as significant reductions in survival, development, and reproduction. Performance of biotype B on AXR#1 was similar to its performance on Cabernet Sauvignon. For both AXR#1 and 5C rootstocks and both phylloxera biotypes A and B the antibiotic mechanism was considerably stronger than the antixenotic mechanism. This research indicated that host preference and performance are positively correlated in grape phylloxera with the rootstocks and phylloxera biotypes tested.