接种番茄斑萎病毒番茄植株对西花蓟马生物学特性的影响

西花蓟马Frankliniella occidentalis(Pergande)是我国的一种重要入侵害虫。本文研究了西花蓟马在番茄3种处理(健康CK、机械接种番茄斑萎病毒MI、机械损伤MD)叶片上的生长发育、存活及种群增长。结果表明:健康、机械接种番茄斑萎病毒、机械损伤叶片上的发育历期依次为12.68、12.99和11.79d。雌雄成虫寿命和雌虫繁殖能力在各处理叶片上差异不显著(P>0.05)。健康、机械接种番茄斑萎病毒、机械损伤叶片上的内禀增长率依次为0.1362、0.1526和0.1292d-1。本研究表明,接种番茄斑萎病毒的番茄叶片未缩短西花蓟马发育历期,也不能延长寿命及提高产卵量,不能明显加速种群数量增长。这意味着番茄斑萎病毒对西花蓟马在番茄叶片上的生物学特性未能产生明显的有利作用。     

番茄斑萎病毒对多杀菌素抗性西花蓟马发育繁殖和药剂敏感性的影响

【目的】西花蓟马Frankliniella occidentalis (Pergande)是重要的入侵害虫,是番茄斑萎病毒（TSWV）最有效的传播媒介,TSWV对西花蓟马的生长发育有一定的影响。多杀菌素是防治西花蓟马最有效的药剂之一,但已有田间西花蓟马对多杀菌素产生抗药性的报道。TSWV对抗性西花蓟马是否也有影响及程度如何尚不清楚。本研究通过对此问题进行深入研究,以期为进一步了解TSWV对西花蓟马的影响提供依据。【方法】应用特定年龄-龄期及两性生命表的方法,研究用番茄斑萎病毒处理和未处理的多杀菌素抗性和敏感西花蓟马种群的生物学特性;用叶管药膜法测定不同处理种群对3种药剂（多杀菌素、甲氨基阿维菌素苯甲酸盐和虫螨腈）的敏感性变化。【结果】对于抗性品系,TSWV处理后西花蓟马的发育历期缩短,雌成虫寿命和产卵量略高,但与对照组差异不显著(P＞0.05),内禀增长率（r）和净生殖率（R₀）分别为0.0433 d^-1和2.210,显著高于对照组（分别为0.0356 d^-1和1.972）（P ≤ 0.001）。对于敏感品系,TSWV处理后西花蓟马的发育历期缩短,雌雄成虫寿命均显著延长（P ≤ 0.001）,产卵量也略有提高,R₀为4.125,显著高于对照组（3.979）（P ≤ 0.001）。TSWV处理后敏感和抗性西花蓟马对多杀菌素的敏感性没有发生明显变化,对甲氨基阿维菌素苯甲酸盐和虫螨腈的敏感性显著降低。【结论】番茄斑萎病毒对多杀菌素敏感和抗性西花蓟马均有直接有利影响,病毒处理的西花蓟马发育历期缩短,繁殖能力增强,成虫寿命延长,对药剂的敏感性降低。     

西藏拉萨地区西花蓟马种群生命表及发生动态研究

【目的】明确拉萨地区西花蓟马Frankliniellaoccidentalis的生物学特性和田间动态规律。【方法】建立了该种群的两性生命表,采用黄板诱集成虫的方法调查了日光温室番茄上西花蓟马的周年发生动态。【结果】该种群西花蓟马成虫前期总发育历期为12.54 d,2龄若虫发育历期为5.62 d,占成虫前期发育历期的44.8%,卵能存活到成虫阶段的概率为0.598。雌虫的平均寿命为27.76 d,雄虫为20.2 d。该种群的内禀增长率(r)和净增殖率(R0)分别为0.153/d和22.817。在拉萨日光温室番茄上西花蓟马能够周年发生,种群增长呈"单峰"型,6月中旬至8月上旬为发生盛期。【结论】拉萨地区西花蓟马的2龄若虫期明显较长,日光温室中西花蓟马的最佳防治适期在4月初。     

二斑叶螨的卵和丝网对西花蓟马生长发育和繁殖的影响

为探讨二斑叶螨Tetranychus urticae Koch的卵和丝网对西花蓟马Frankliniella occidentalis(Pergande)的影响,本文研究了西花蓟马分别取食健康菜豆叶片、带二斑叶螨卵的菜豆叶片、带卵+丝网的菜豆叶片3种不同情况下,西花蓟马生长发育、存活和繁殖情况,并获得了其生命参数。结果表明西花蓟马在带二斑叶螨卵的菜豆叶片上发育最快,其未成熟期为11.35 d,明显短于另外两个处理,在带有二斑叶螨卵和丝网的菜豆叶片上成虫获得率最低为63%,西花蓟马在不同处理之间的成虫寿命和繁殖力没有明显的差异;西花蓟马取食3种菜豆叶片的内禀增长率分别为0.1779、0.1827和0.1710。以上结果说明取食二斑叶螨的卵可缩短西花蓟马发育时间,对种群增长有利,二斑叶螨的丝网可降低西花蓟马的存活率。     

西花蓟马与烟蓟马生物学特性的比较研究

本试验研究并比较了外来入侵物种西花蓟马Frankliniella occidentalis(Pergande)与本地物种烟蓟马Thrips tabaci Lindeman的生物学特性,包括种群增长速度、发育历期、成虫寿命、孤雌生殖方式下的产卵量以及产卵规律等几个方面。结果显示,当以相同的数量在紫甘蓝上饲养时,只经过3代,烟蓟马的种群数量就可以达到西花蓟马的3倍,随后对2种蓟马种群的性别比进行了鉴定后发现,西花蓟马种群的雌雄比为2.8∶1,而烟蓟马种群中的所有个体均为雌性。除了2龄若虫期和预蛹期之外,烟蓟马在紫甘蓝上的发育历期明显长于西花蓟马,而其雌虫的平均寿命显著短于西花蓟马雌虫,分别是24.94d和43.35d。进行孤雌生殖时,西花蓟马的终生产卵量明显多于烟蓟马,但是在日均产卵量方面,这一情况却相反。     

西花蓟马酵母双杂交文库及TSWV膜蛋白诱饵载体的构建

【目的】为筛选西花蓟马Frankliniella occidentalis与番茄斑萎病毒的互作蛋白。【方法】利用MakeYour Own"MatePlate?"Library System构建西花蓟马酵母双杂交初级cDNA文库以及Y187酵母次级文库,利用Matchmaker Gold Yeast Two-Hybrid System构建TSWV膜蛋白GN及GC诱饵载体。【结果】建立的西花蓟马初级cDNA文库库容为5.6×106 cfu,其插入片段平均长度大于1 000 bp,重组率约为100%。建立的Y187酵母次级文库转化效率为5×106cfu/μg,文库滴度为2.97×108。构建的pGNKT7-GN及pGBKT7-GC诱饵载体能够在Y2HGold酵母菌株中正确表达,无自激活活性且无毒性。【结论】成功构建了西花蓟马酵母双杂交文库以及番茄斑萎病毒膜蛋白诱饵载体,可用于后续筛库实验。     

乙基多杀菌素多代胁迫对西花蓟马解毒酶活性、发育和繁殖的影响

【目的】乙基多杀菌素是防治蔬菜上西花蓟马Frankliniella occidentalis的首选药剂之一。本研究旨在明确乙基多杀菌素多代胁迫对西花蓟马的影响。【方法】采用浸叶法以LC₂₅浓度的乙基多杀菌素对西花蓟马2龄若虫连续汰选6代后,测定其抗药性和解毒酶活性的变化,并用年龄-阶段两性生命表分析LC₂₅浓度乙基多杀菌素对西花蓟马后代(F₇)生命表参数的影响。【结果】LC₂₅浓度的乙基多杀菌素对西花蓟马2龄若虫连续汰选6代后,与用清水处理的对照相比,抗性倍数达到了6.4倍;F₆代2龄第2天若虫体内羧酸酯酶(CarE)和谷胱甘肽-S转移酶(GSTs)被激活,活性分别增加至对照的1.26和1.21倍;而多功能氧化酶(MFOs)和细胞色素P450酶(P450s)受到明显的抑制,活性分别只有对照的54.00%和51.78%。LC₂₅浓度的乙基多杀菌素胁迫6代不仅造成西花蓟马雌成虫寿命(19.33 d)和单雌产卵量(69.80粒)较对照(分别为21.60 d和83.17粒)显著下降,且后代(F₇)的雌成虫寿命(16.82 d)和雄成虫寿命(8.29 d)较对照(分别为20.28 d和10.86 d)极显著缩短,单雌产卵量(59.23粒)较对照(76.96粒)极显著降低,但雌雄比(2.28)较对照(1.03)显著提高,种群生命参数净增殖率(R₀)、内禀增长率(r)和周限增长率(λ)则无明显变化。【结论】在LC₂₅浓度乙基多杀菌素连续汰选6代内西花蓟马对该药剂的抗性缓慢增长;药剂胁迫不仅对西花蓟马解毒酶活性有不同程度的影响,还对其发育和繁殖具有一定的抑制作用。     

吡虫啉胁迫对西花蓟马生长发育及种群性比的影响

为明确吡虫啉持续大量使用对外来入侵害虫西花蓟马Frankliniella occidentalis的影响。本研究通过毒力测定得到西花蓟马室内种群LC_(30)及LC_(50)的两种吡虫啉浓度162 mg/L和171 mg/L,在室内人工气候箱环境下研究了西花蓟马各虫态的发育历期、成虫寿命及性比等相关种群参数。结果表明,西花蓟马未成熟虫期的发育历期在经过吡虫啉处理后相比对照显著缩短(P0.05),且雌性西花蓟马的未成熟虫期明显短于雄性西花蓟马,其中以171 mg/L吡虫啉处理后西花蓟马的雌性种群未成熟虫期发育历期缩短最为明显(P0.05);西花蓟马雄性种群的寿命略长于雌性种群,且171 mg/L吡虫啉处理后雌雄虫寿命与对照相比均显著缩短(P0.05);吡虫啉处理后西花蓟马的平均产卵期低于对照,但单雌产卵量、日均产卵量均显著高于对照,171 mg/L吡虫啉处理后的单雌产卵量和日均产卵量最高(P0.05)。吡虫啉胁迫后,西花蓟马的雌性种群在整个种群中所占比例增速迅速上升,经过125 d(大约5代)之后,雌性种群已经占绝对优势,几乎取代雄性种群,即种群中90%以上均为雌性西花蓟马。西花蓟马雌性种群对吡虫啉的胁迫适应性强于雄性种群,吡虫啉胁迫是造成西花蓟马雌性种群比例迅速升高的主要原因之一,在吡虫啉浓度LC_(30)到LC_(50)胁迫的范围内,随着吡虫啉胁迫浓度的增加,种群中雌性种群所占比例增大。     

辣椒素对西花蓟马雌成虫体内解毒酶活性的影响

【目的】为明确非挥发性次生物质辣椒素对西花蓟马Frankliniella occidentalis雌成虫体内解毒酶活性的影响。【方法】将菜豆叶片研碎离心后取上清液作为西花蓟马的液态饲料,将不同浓度的辣椒素添加到饲料中,饲养西花蓟马雌成虫48、72、96 h后,分别测定西花蓟马体内羧酸酯酶(Car E)、谷胱甘肽-S-转移酶(GSTs)、乙酰胆碱酯酶(AChE)和多功能氧化酶(MFO)的活性。【结果】低剂量辣椒素能诱导西花蓟马雌成虫体内GSTs、AChE和MFO活性明显升高,高剂量辣椒素抑制GSTs、AChE及MFO活性。不论在哪一种辣椒素浓度下,Car E活性一直受到抑制。辣椒素浓度和处理时间对GSTs及AChE活性无明显交互作用,但对Car E及MFO有明显的交互作用,处理时间越长,作用越显著。【结论】西花蓟马雌成虫体内解毒酶活性在次生物质辣椒素处理下发生了明显的变化,说明西花蓟马通过解毒酶适应次生物质对其的影响,但解毒酶的变化有剂量和时间效应。     

CO_2浓度升高对不同寄主植物上西花蓟马和花蓟马成虫体内消化酶活性的影响

【目的】为明确大气CO_2浓度升高对入侵昆虫西花蓟马Frankliniella occidentalis及其本地近缘种花蓟马F.intonsa的影响机制。【方法】在CO_2人工气候箱(400和800μL/L)内分别以不同的寄主植物(四季豆、茼蒿、辣椒和黄瓜)饲养西花蓟马和花蓟马3代以后,比较分析两种蓟马成虫体内消化酶(淀粉酶、胰蛋白酶和脂肪酶)活性的差异。【结果】4种寄主植物饲养的西花蓟马和花蓟马成虫体内淀粉酶活性均随着CO_2浓度的升高而降低,其中四季豆、黄瓜、辣椒和茼蒿饲养的西花蓟马成虫体内淀粉酶活性依次降低21.39%,25.33%,44.59%和42.27%;而花蓟马成虫体内淀粉酶活性则分别降低了48.79%,49.47%,38.86%和38.92%。随着CO_2浓度的升高,两种蓟马成虫体内脂肪酶和胰蛋白酶的活性均升高,其中四季豆、黄瓜和辣椒饲养的两种蓟马酶活性均显著升高(P0.05)。高CO_2浓度环境中四季豆饲养的西花蓟马脂肪酶和胰蛋白酶活性分别是对照的2.00和2.49倍,黄瓜饲养的种群其脂肪酶和胰蛋白酶活性分别是对照的2.36和2.27倍,辣椒饲养种群相应的酶活性分别是对照的3.61和3.59倍。而高CO_2浓度环境中四季豆、黄瓜和辣椒饲养的花蓟马脂肪酶活性仅分别是对照的1.76,2.18和2.69倍,胰蛋白酶活性分别为对照的1.72,2.19和2.42倍。【结论】CO_2浓度升高和寄主植物种类是影响两种蓟马成虫体内淀粉酶、胰蛋白酶和脂肪酶活性变化的重要原因,两种蓟马均能通过调节自身的酶活性来适应未来高CO_2浓度的气候环境,其中入侵种西花蓟马对环境的适应性强于本地近缘种花蓟马。     

Influence of tomato spotted wilt virus on performance and behaviour of western flower thrips (Frankliniella occidentalis)

The general principles in pathogen transmission by insects involve a complex and specific interplay, in this case between thrips, tospovirus and their shared host plant, which has led to outbreaks of crop disease epidemics of economic and social importance. The possible processes and factors driving their co‐evolution were partly studied by rearing Frankliniella occidentalis [western flower thrips (WFT)] on either tomato spotted wilt virus (TSWV)–infected or uninfected Capsicum annum leaflets throughout their larval stages. Later, pupae were transferred individually on healthy leaf discs for further studies of the influence of TSWV on WFT development and behavioural patterns. The exposure of WFT to TSWV was found to improve performance with regard to longevity and survival, with mean longevity being significantly higher in TSWV‐exposed WFT compared to unexposed ones (F_(3,403) = 22.44, P < 0.0001). The observed improvement in survival was as a result of significant reduction in mortality for the WFT individuals exposed to TSWV (F_(3,383) = 849.94, P < 0.0001) compared to the unexposed. However, the results showed a significant reduction in mean daily fecundity overtime (F_10,10) = 246.66, P < 0.0001) and across the four treatments (F_(3,30) = 6.62, P = 0.001), as well as lifetime fecundity (F_(3,23) = 21.23, P < 0.0001) of the WFT exposed to TSWV compared to the unexposed reared on uninfected leaf discs. For preferential test, C. annum leaf discs infected with TSWV were more attractive to WFT as compared to healthy leaf discs (χ²_{(4, 34)} = 112.35, P < 0.0001). These results are envisaged to contribute to a clear understanding into the plant–vector–virus interaction, which is essential for accurate diagnosis and control of the TSWV epidemic, as well as the control of F. occidentalis as crop pest.     

The incidence and distribution of viruses infecting lettuce, cultivated Brassica and associated natural vegetation in Spain

A virus survey was conducted during the spring and autumn of 2001 and 2002 to determine the presence, prevalence and distribution in Spain of the viruses that are most commonly found infecting lettuce and Brassica worldwide. Crop plants showing virus symptoms from the principal lettuce and Brassica-growing regions of Spain, and some samples of the annual and perennial flora nearby, were tested by enzyme-linked immunosorbent assays using specific commercial antibodies against the following viruses: Alfalfa mosaic virus (AMV), Broad bean wilt virus 1 (BBWV-1), Beet western yellows virus (BWYV), Cauliflower mosaic virus (CaMV), Cucumber mosaic virus (CMV), Lettuce mosaic virus (LMV), Pea seed-borne mosaic virus (PSbMV), Turnip mosaic virus (TuMV) and Tomato spotted wilt virus (TSWV). Samples were also tested with a Potyvirus genus antibody. Virus incidence was much lower in spring than in autumn, especially in 2001. In spring 2002, CMV and LMV were the most prevalent viruses in lettuce, while CaMV was the most important virus present in Brassica crops grown in Navarra, followed by CMV and BWYV. In the autumn, the spectrum of viruses was different; potyviruses were widespread in lettuce grown in Madrid, but TSWV and BWYV were predominant in the Murcia region. The prevalent Potyvirus detected in lettuce fields was LMV, but none of the samples collected were positive for PSbMV or TuMV. In Brassica crops, TSWV was the most abundant in autumn-sown crops, especially in the Navarra region. All of the viruses present in lettuce and Brassica were also frequently detected in their associated natural vegetation at the same time, suggesting that they probably play an important role as virus reservoirs. Sonchus spp. were particularly common and were frequently infected with CMV, LMV and BWYV. Another common species, Chenopodium album, was often infected with TSWV and BWYV. Multiple infections were common, especially in non-crop plants, and the most common combination was BWYV and TSWV. The role of weeds in the epidemiology of viruses that infect lettuce and Brassica crops in Spain is discussed.     

Thrips tabaci and tomato spotted wilt virus: inheritance of vector competence

Populations of onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), were shown to differ significantly in their ability to transmit an isolate of tomato spotted wilt virus (Tospovirus: Bunyaviridae) (TSWV) collected from potato [Solanum tuberosum L. (Solanaceae)]. To gain an understanding of the basis for this variation, we generated reciprocal crosses between an efficient and an inefficient transmitting population. The resulting F₁ progeny and progeny from the parental populations were tested for their ability to transmit TSWV. Our results indicate that the ability to transmit TSWV efficiently by T. tabaci is inherited as a recessive trait.     

Влнянне характера эаселення ноля капустноii совкоii(Barathra brassicae L.) эффективноеть трихограммы (Trichogramma evanescens Westw.) (Lepidoptera,Noctuidae; Hymenoptera,Trichogrammatidae)

TSWV belongs to the genus Tospovirus which was established in the family Bunyaviridae, a family of animal viruses. Besides TSWV, Impatiens necrotic spot virus (INSV) and ground nut bud necrosis virus (GBNV) were established as different Tospovirus species. Tospoviruses have quasispherical particles of 85 nm diametre which are surrounded by a membrane and contain 3 RNA species and 4 structural proteins. In Tospovirus infected plant cells virions were detected in cavaties of the endoplasmatic reticulum and additionally amorphous electron dense material accumulates in infected cells. Defective forms of TSWV lack the ability to form complete virus particles. TSWV is the only plant pathogenic virus that is transmitted by thrips which transmit the virus with different efficiency. The virus has an extensive plant host range of more than 360 different species. The developing symptoms depend on the Tospovirus species, the virulence of the virus strains and the environmental conditions.

Based on the reaction of TSWV isolates with N‐specific polyclonal antisera, 3 serogroups were established. The most frequently used technique for serologically based diagnosis of Tospoviruses is DAS ELISA with N‐specific or preadsorbed antisera against complete virus. For TSWV epidemiology distinct weeds and cultural host plants play an important role for the survival of virus and vector. Breeding for resistance is the most important preventive measure of control.     

A Survey of Tospoviruses in Bulgaria

A survey of tospoviruses present in Bulgaria was conducted during three vegetation periods. A total of 258 different virus isolates were obtained from tobacco, tomato, weeds and ornamentals in the open fields and from glasshouses. Virus isolates were identified by inoculation to test plants and serology as Tomato spotted wilt virus (TSWV). No indications were obtained that other Tospovirus species were present in Bulgaria. In addition the transmission ability and efficiency of several Bulgarian populations of Thrips tabaci (Lind.) and Frankliniella occidentalis (Perg.) was determined. The experimental results indicated that Bulgarian TSWV isolates are transmitted with high efficiency by F. occidentalis and that only arrhenotokous and not thelytokouse populations of T. tabaci could transmit TSWV.     

Serological relationships and purification of bud necrosis virus,a tospovirus occurring in peanut (Arachis hypogaea L.) in India*

A procedure for the purification of a tospovirus which causes bud necrosis disease (BND) of peanut in India is described. The virus contained three polypeptides of 78 kDa, 54 kDa and 31 kDa. In two ELISA procedures the virus failed to react with antisera to tomato spotted wilt virus (TSWV) obtained from different sources and with an antiserum to impatiens necrotic spot virus (INSV). Additionally, in reciprocal tests TSWV and INSV antigens failed to react with antiserum to the virus infecting peanut in India. In electro-blot immunoassay 54 kDa and 31 kDa polypeptides of the virus reacted with the homologous antiserum. None of the heterologous antisera reacted with any of the three viral polypeptides. On the basis of serological differences the virus that causes BND in India is distinct and therefore has been named bud necrosis virus (BNV). This serotype appears to be restricted to Asia.     

我国南方花生发生一种由蕃茄斑萎病毒引起的新病害

我们于1984和1985年6月上、中旬,在广州市郊、县,湛江市郊以及广西南宁市郊、县,北海市郊和合蒲县等花生产区,调查花生病毒病时,除了发现花生轻斑驳病毒病外,还发现一种新的病毒病害。其症状特征是:病株顶端叶片上出现很多褪绿黄斑或环斑,有的环斑变