RAPD方法用于区分中国烟粉虱的生物型

应用RAPD－PCR技术研究了中国大陆10个烟粉虱种群的生物型情况,聚类分析表明,这10个种群中包含了3种生物型：烟粉虱B型,广西型（南宁南瓜寄主）和福建型（福州地瓜）。值得注意的是,在福建并存着两种烟粉虱生物型,即苷蓝上的B型烟粉虱和甘署上的福建福州型,研究表明烟粉虱B型已经传入中国,至少在北京,山东,新疆,福建,广东等地都有发生和分布,广西型和福建型是否中国的本地土种群还需进一步研究。     

利用mtDNA COⅠ基因序列鉴定我国烟粉虱的生物型

利用mtDNA COⅠ基因片段作标记,采用序列分析的方法,从分子生态学的角度研究了近年来在我国暴发危害的烟粉虱Bemisia tabaci 5个种群（北京一品红种群,广州甘蓝种群,西安一品红种群,北京西红柿种群和新疆吐鲁番棉花种群）的生物型。在5个种群的基因片段上,截取与Texas-B型相应的720bp的序列分析,结果表明所测序列中只有2个碱基与Texas-B型不同,序列相似性为99.7%;在西安一品红种群和新疆吐鲁番棉花种群的原序列中截取与Arizona-B型序列相应的423bp片段,分析表明这两个种群与AZB3型属于同一个单倍型。因此,我国烟粉虱5个实验种群的生物型与Texas-B型和Arizona-B型种群为同一生物型“B”。     

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。     

烟粉虱生物型研究进展

烟粉虱B em isia tabaci(G ennad ius)是一种取食植物汁液的重要农业害虫,同时也是许多植物病毒最重要的传播介体之一。烟粉虱被认为是由许多具有明显遗传分化的不同种群即生物型组成的复合种,其中B型烟粉虱是一种入侵性最强的生物型,几乎在世界各地均有分布。由于不同生物型的烟粉虱在寄主范围、传毒能力、地理分布、抗药性等许多生物学方面存在差异,因此对烟粉虱生物型的鉴定及其遗传分化研究对于该害虫的可持续控制具有重要的指导意义。烟粉虱生物型的鉴定方法包括生物学鉴定、酶谱鉴定以及分子标记鉴定的方法,其中使用的分子标记包括RAPD、AFLP、rDNA-ITS1、m tDNA CO I以及SSR等标记。目前,不同生物型的烟粉虱尤其B型烟粉虱的分类地位仍然存在争议。介绍了不同生物型烟粉虱的生物学差异、鉴定方法及其遗传分化研究的最新进展,同时探讨了不同生物型烟粉虱的分类地位和研究方向。     

江苏烟粉虱生物型鉴定与危害程度分析

通过实地调查结合mtDNA COI基因测序方法,分析并鉴定了江苏地区主要作物上烟粉虱的危害程度及烟粉虱的生物型组成。序列检测表明,江苏地区烟粉虱种群生物型为Q型和B型,其中Q型烟粉虱占所测总样本94.40%,Q型烟粉虱在苏南、苏中和苏北的比例分别是97.5%、96.0%和91.67%,表明Q型烟粉虱已成为在江苏省主要生物型烟粉虱。同源性分析发现,江苏的Q型烟粉虱序列与浙江、美国和日本的Q型烟粉虱碱基序列相似性为100%,江苏的B型烟粉虱序列与巴基斯坦、日本和中国北京的B型烟粉虱序列同源性为100%。在江苏地区烟粉虱主要危害茄科的番茄、长茄、甜椒,葫芦科的黄瓜、西葫芦及锦葵科的苘麻、棉花,表现出虫口密度大,植株受害重,而十字花科的小白菜、大白菜、萝卜及豆科的菜豆、豇豆等受害较轻,苋科的苋菜和菊科的生菜受害最轻。     

江苏地区烟粉虱生物型演替研究初报

本文连续5年系统监测了江苏省13个地级市烟粉虱Bemisia tabaci(Gennadius)生物型的发生分布状况,探讨了它们的迁移扩散和演替规律。利用RAPD分子标记和mtDNA COI基因序列进行烟粉虱生物型鉴定,结果表明:江苏地区发生的烟粉虱生物型为B型和Q型。自2005年到2009年,B型烟粉虱在苏南和苏北地区的发生分布频率逐年下降,在苏北地区的发生分布频率由52.58%下降为22.22%,在苏南由56.52%下降为9.37%;而Q型烟粉虱在苏南和苏北的发生分布频率逐年升高,在苏北由47.42%上升至77.78%,在苏南由43.48%上升为90.63%。B型烟粉虱的发生分布范围由全省逐渐向苏北地区缩小,而Q型烟粉虱的发生分布范围逐渐扩大遍及全省;此外,Q型烟粉虱在江苏是由苏中和苏南地区向苏北地区扩散,并逐步取代B型烟粉虱成为江苏地区农作物的主要害虫。     

云南Q型烟粉虱种群的鉴定

利用mtDNACOⅠ基因片段作标记,对采自云南昆明一品红上的烟粉虱Bemisiatabaci(Gennadius)种群生物型进行了鉴定。结果表明,该种群COⅠ基因片段(501bp)与摩洛哥Q型、西班牙Q型烟粉虱的相应碱基序列具有极高的同源性,与2种Q型烟粉虱的COⅠ基因仅分别相差3个和4个碱基,同源性分别达到994%和992%。序列分析表明我国云南昆明一品红上存在Q生物型烟粉虱     

国内烟粉虱B生物型的分布及其控制措施研究

收集了国内 15个省 2 6个不同地区的烟粉虱种群 ,根据有关报道合成了一个随即引物H16 ,对不同地区的烟粉虱DNA进行了RAPD -PCR扩增 ,根据扩增结果分析了不同烟粉虱种群的生物型。结果表明 ,在以上不同省市和地区中 ,北京、山东等 11个省 19个地区都有烟粉虱B型的分布。以烟粉虱B型为防治对象 ,比较了使用高效氯氰菊酯、高效大功臣 2种化学农药 ,和应用桨角蚜小蜂、粉虱座壳孢 2种寄生性天敌 ,对黄瓜上烟粉虱种群的控制效果 ,结果表明在一个世代内 ,喷施高效大功臣 (10 0 0× ,1次 ) ,喷施粉虱座壳孢 (5× 10 6 个孢子 /ml,2次 ) ,和释放桨角蚜小蜂 (3头雌蜂 /株 ,2次 ) ,对烟粉虱种群的控制作用分别达到 78.2 1%、95 .74 %和 5 7.5 8% ,三者都可以对烟粉虱起到较好的防治效果。     

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。     

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。     

Development and reproduction of ‘B＇ biotype Bemisia tabaci （Gennadius） （Homoptera： Aleyrodidae） on four ornamentals

Abstract Effects of four commercial ornamentals on the development, survivorship and reproduction of the whitefly B biotype Bemisia tabaci (Gennadius) were studied in the laboratory (temperature 26 ± 1 °C; relative humidity 75%–90%; L: D 14:10). The total survivorship from egg to adult on hibiscus ( Hibiscus rosa-sinensis L.), poinsettia ( Euphorbia pulcherrima Will), cottonrose hibiscus ( Hibiscus mutabilis L.) and variegated leafcroton ( Codiaeum variegatum'Aucubaefolium' ) were 33.69%, 40.55%, 79.11%, and 29.39%, respectively. The developmental periods from egg to adult varied from 23.12 days on cottonrose hibiscus to 32.13 days on hibiscus. The average longevity of adult females ranged from 6.87 days on variegated leafcroton to 21.07 days on poinsettia. The average numbers of eggs laid per female were 9.20, 25.13, 54.45, and 26.79 on the above respective hosts. The intrinsic rates of natural increase ( r_m ) for B biotype B. tabaci on cottonrose hibiscus was the highest. Based on life table analyses of whitefly populations, cottonrose hibiscus was the most suitable host for B biotype B. tabaci in this study.     

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。     

Rapid identification of B biotype of Bemisia tabaci (Homoptera: Aleyrodidae) based on analysis of internally transcribed spacer 1 sequence

Abstract B biotype is a reasonably important biotype among all known biotypes in the Bemisia tabaci species complex. Local populations of B. tabaci on different host plants were collected from across the Chinese mainland, Taiwan, Pakistan and Israel. From each population of B. tabaci , an internally transcribed spacer region of the ribosomal rDNA gene was amplified, cloned and the sequence determined. Sequence homology analyses were performed and the results were similar to those based on morphology and biological characters. Based on analysis of the internally transcribed spacer 1 sequences, a B biotype-specific primer was designed. The PCR diagnosis results showed that B biotype is identifiable by a specific PCR product by using the forward diagnostic primer paired with a universal reverse primer. This diagnostic primer-based protocol can be used for preliminary analysis of mixed Bemisia populations containing B biotype, as well as other biotypes.     

不同寄主植物对烟粉虱发育和繁殖的影响

研究了烟粉虱 B型 ( Bemisia tabaci Gennadius)在番茄、茄子、黄瓜和甘蓝上的发育、存活和繁殖情况。在 2 6± 1℃的条件下 ,烟粉虱从卵发育到成虫的存活率以在甘蓝上的最高 ,为 68.5 5 % ,黄瓜上的最低 ,为46.2 8% ;发育时间以在茄子上最短 ,为 1 7.5 d,黄瓜上最长 ,为 1 9.3d,差异显著 ;平均单雌产卵量以在甘蓝上最大 ,为 1 43粒 ,黄瓜上最小 ,为 98.2 5粒 ;成虫的寿命以在甘蓝上最长 ,平均为 2 5 .2 d,黄瓜上为 1 7.2 d;内禀增长率 rm以在茄子上的最大 ,为 0 .1 41 6,黄瓜上最小 ,为 0 .1 1 43。综合比较 4种不同寄主植物 ,茄子是烟粉虱种群生长发育和繁殖最适宜的寄主     

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。     

Distribution and molecular characterization of distinct Asian populations of Bemisia tabaci (Hemiptera: Aleyrodidae) in Japan

Bemisia tabaci (Gennadius) is considered to be the most economically important pest insect worldwide. The invasive variant, the Q biotype of B. tabaci was first identified in 2004, and has caused significant crop yield losses in Japan. The distribution and molecular characterization of the different biotypes of B. tabaci in Japan have been little investigated. In this study, B. tabaci populations were sampled from the Japanese Archipelago, the Amami Archipelago and the Ryukyu Islands between 2004 and 2008, and the nucleotide sequences of their mitochondrial cytochrome oxidase I genes were determined. Bayesian phylogenetic relationship analysis provided the first molecular evidence that the indigenous Japanese populations could be separated into four distinct genetic groups. One major native population from the Japanese Archipelago, given the genetic group name Lonicera japonica, was separated into an independent group, distinct from the other genetic groups. The second major population, the Nauru biotype in the Asia II genetic group, was identified in the Amami Archipelago and the Ryukyu Islands. Two distinct minor genetic groups, the Asia I and the China, were also identified. One invasive B‐related population belonging to the Mediterranean/Asia Minor/Africa genetic group has been identified in Honshu. All lineages generated by the phylogenetic analyses were supported by high posterior probabilities. These distinct indigenous B. tabaci populations developed in Japan under geographical and/or biological isolation, prior to recent invasions of the B and Q biotypes.     

寄主作物对B型烟粉虱生长发育和种群增殖的影响

在恒温28℃、湿度80％的条件下研究了5种寄主植物对B型烟粉虱(Bemisia tubaci)生长、发育和繁殖的影响。结果表明：取食棉花、烟草、花生、大豆和玉米的烟粉虱。其形态、发育历期、存活率、成虫繁殖力和寿命等生命参数均有明显差异。棉花、大豆和玉米不利于烟粉虱卵和若虫的生长发育。表现为虫体较小、卵期和生命周期延长、存活率降低。烟粉虱在棉花、大豆、花生、烟草和玉米上的生命周期分别为32．03d、32．11d、25．69d、24．43d和20．68d。其存活率分别为49．86％、54．41％、86．86％、69．93％和29．38％。与烟草和玉米相比。棉花、大豆和花生明显有利于烟粉虱的种群繁殖．在棉花、大豆、花生、烟草和玉米上生长发育的成虫寿命和单雌产卵量分别是27．8d和235．O粒、23．2d和191．1粒、22．Od和131．1粒、6．25d和28．O粒、2．42d和5．1粒。在花生、大豆、棉花、烟草和玉米上的内禀增长率(rm)分别为O．1590、O．1364、O．1236、O．0841和-O．0285．其种群趋势指数(I)分别为113．85、117．38、103．98、19．58和O．4274。     

Biotype and status of insecticide resistance of whitefly Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) in Alhassa oasis,Eastern Province of Saudi Arabia.

The tobacco whitefly, Bemisia tabaci, is one of the major insects infesting vegetable plants. This pest is well known in Alhassa oasis, Saudi Arabia; which is historically agricultural land cultivated with date palm trees and different vegetables. A molecular key based on the sequence of the mitochondrial cytochrome oxidase gene CO1 was used for the identification of strains of the tobacco whitefly Bemisia tabaci collected from farms located in four areas of the Alhassa oasis; Northern, Southern, Eastern and Western. Only one biotype (B‐biotype) of B. tabaci was reported in the oasis. Resistance of B. tabaci was tested against eight insecticides, the results showed moderate to low levels of resistance to the tested insecticides. However, the resistance to neonicotinoid insecticides was low and established at 1.3 fold to both Imidacloprid and Acetamiprid. In addition, medium levels of resistance were detected to the insect growth regulator Pyriproxyfen (30 fold), and the pyrethroid Deltamethrin (30 fold), Bifenthrin (24 fold) and Cypermethrin (13 fold). A medium level of resistance was also detected to the carbamate insecticide Carbosulfan and was 40 fold of the laboratory strains. A low level of resistance to the organophosphorus insecticide was detected to Phenthoate (11 fold). However, these results reflect that the farmers were less dependent on the use of insecticides to control B. tabaci in the oasis and they may be implementing other environmentally sound techniques to keep the pest below the damage threshold.     

Development and reproduction of Bemisia tabaci （Homoptera： Aleyrodidae） on three bean species

Abstract The development, survivorship and reproduction of Bemisia tabaci B-biotype on three bean species were studied at 26±1°C in the laboratory. The developmental periods from egg to adult varied from 27.80 days on garden beans to 18.20 days on soybeans. The survivorship from egg to adult on soybeans, cowpeas and garden beans was 77.14, 70.14 and 64.28%, respectively. The average longevity of female adults ranged from 12.30 days on soybeans to 9.80 days on garden beans, and the oviposition of B. tabaci varied from 160.85 eggs on soybeans to 98.00 eggs on garden beans. Life table parameters were calculated as biological attributes for Bemisia tabaci populations fed on three bean species. The results indicated that the intrinsic rate of increase (r_m), the finite rate of increase (λ) and net reproductive rate (R₀) were high for populations fed on soybeans, with values of 0.1857, 1.2041 and 82.1576, respectively. The corresponding values were less for populations fed on garden beans, with values of 0.1097, 1.1159 and 31.2661, respectively. The parametric values for cowpeas were intermediate between soybeans and garden beans but no significant difference were observed for the r_m values for soybeans and cowpeas. Experimental evidence in our investigation indicated that Bemisia tabaci is best adapted and shows the greatest preference for soybean of the three bean species tested in this study.