基于TaqMan实时荧光定量PCR技术的西花蓟马快速检测

西花蓟马Frankliniella occidentalis（Pargande）是世界性害虫,2003年在我国首次发生危害。针对西花蓟马与其他种类蓟马形态相似、难以快速区分的问题,本文在SCAR标记基础上,采用TaqMan实时荧光定量PCR技术,设计1对特异性引物和1条MGB探针,扩增出大小为138bp的特异片段。以质粒DNA为标准品建立了标准曲线（R2=0.9965）,种特异性检验结果显示,该引物和探针只能检测到西花蓟马的荧光信号,而对其他种类的蓟马不具有检测能力。并且可以定量检测西花蓟马不同虫态靶标DNA片段的拷贝数。该检测体系重复性强、稳定性高,在口岸检疫以及植物种苗及其产品调运中的有害生物检测和监测中具有重要意义。     

西花蓟马和花蓟马在温度逆境下的存活率比较

以外来入侵害虫西花蓟马(Frankliniella occidentalis)和本地种花蓟马(F.intonsa)为对象,研究了高温和低温逆境下两种蓟马各虫态存活率的差异.结果表明:在36℃～44℃高温下暴露2h和4h,不论是西花蓟马还是花蓟马,各虫态的存活率均随温度的升高而降低,但在相同高温处理条件下,同一虫态西花蓟马的存活率要高于花蓟马;在-2℃～-10℃低温下暴露2h和4h,西花蓟马和花蓟马各虫态的存活率均随随温度的降低而下降;在相同低温处理条件下,同一虫态西花蓟马的存活率也要高于花蓟马.由此表明,2种蓟马对高温和低温的适应能力不同,本地种花蓟马对高温和低温均比较敏感,而外来入侵害虫西花蓟马对极端温度具有较强的适应性.     

外来入侵害虫西花蓟马防控技术研究与示范

针对入侵害虫西花蓟马Frankliniella occidentalis(Pergande)在我国部分地区暴发成灾的形势,公益性行业科研专项"外来入侵害虫西花蓟马防控技术研究与示范"项目组在除西藏、台湾、香港和澳门以外的30个省、区和直辖市开展了西花蓟马的调查、监测、预警和综合防控技术研究与示范。结果表明西花蓟马在北京、云南、浙江、山东等14个省市发生危害,尤以云南和北京两地最为严重,在全国呈快速蔓延趋势。明确了西花蓟马在我国的成灾机制;建立了西花蓟马高效诱捕技术;筛选出4种对西花蓟马具有显著控害潜力的本土天敌昆虫及病原微生物;筛选出5种对西花蓟马高效、对环境友好的防治药剂;明确西花蓟马重要发生区域北京和云南种群对主要化学药剂的抗性和机制;分别组建了基于农业措施防治、色板与引诱剂应用的引诱技术、生物防治和高效环保化学农药综合应用的10套西花蓟马防控技术体系,在我国北京、云南、山东和浙江等西花蓟马主要发生区域累计推广面积2.65万公顷,有效控制了西花蓟马的为害和扩散蔓延。     

蔬菜花期蓟马的种群动态与空间分布研究

西花蓟马是近年来在我国局部地区暴发成灾的重要外来入侵害虫,有关西花蓟马入侵对本地蓟马种群动态、空间分布及优势种影响的报道较少。对云南省昆明市近郊蔬菜花期的蓟马种群动态和空间分布研究表明,蔬菜上的蓟马种类主要是西花蓟马Frankliniella occidentalis(Pergande)、花蓟马F.intonsa(Trybom)、棕榈蓟马T.palmi(Karny)和端大蓟马Megalurothrips distalis(Karny);不同蔬菜上的蓟马优势种存在一定差异,其中辣椒和茼蒿上的蓟马优势种为西花蓟马;韭菜、茄子和四季豆上的蓟马优势种分别为花蓟马、棕榈蓟马和端大蓟马。各蔬菜上的蓟马种群数量以花期为多,盛花期达最大值,其中茄子花上的蓟马成虫平均虫口密度最高,为14.93头/朵。利用聚集度指标进行空间分布检测表明,不同蔬菜上蓟马成虫的空间分布型均为聚集分布,且聚集程度随密度的增加而增大。本研究可为深入探讨西花蓟马对本地蓟马的竞争取代机制积累资料,同时为西花蓟马的综合治理奠定理论基础。     

入侵害虫西花蓟马的生物学、危害及防治技术

西花蓟马Frankliniellaoccidentalis最早发生于北美地区,现已广泛分布于世界各地,是园艺作物最重要的害虫之一。早在1996年,西花蓟马就被我国农业部列为进境植物检疫潜在的危险性害虫,2003年夏季西花蓟马在北京部分保护地首次发现。该文从分布及危害、寄主植物、生物学特性和防治方法4个方面对该害虫进行了概述。     

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。     

花蓟马雄虫释放的聚集信息素的分离和鉴定

明确花蓟马Frankliniella intonsa雄虫释放的聚集信息素对于花蓟马生物防治具有重要意义。但目前除了西花蓟马F. occidentalis, 关于其他蓟马雄虫释放的聚集信息素及其成分鉴定的研究未有报道。本研究采用Y型嗅觉仪测定花蓟马雌雄成虫和西花蓟马雌雄成虫对花蓟马雄虫和西花蓟马雄虫气味源的嗅觉反应; 用固相微萃取（solid-phase microextraction,SPME）方法提取花蓟马雄虫挥发物, 利用含有BGB-176 SE或CP-chirasil-Dex CB 2种不同手性柱的气-质联用仪（gas chromatography-mass spectrometry, GC-MS）对挥发物进行分离和鉴定。行为学试验结果表明： 花蓟马雄虫气味源对其雄性及雌性成虫均具有明显的吸引作用。经GC-MS分析, 花蓟马雄虫挥发物中含有2种主要的化合物, 分别为(R)-lavandulyl acetate和neryl (S)-2-methylbutanoate, 其与已报道的西花蓟马雄虫释放的聚集信息素成分相同。但定量分析结果表明, (R)-lavandulyl acetate与neryl (S)-2-methylbutanoate在花蓟马和西花蓟马雄虫中释放的比例不同; 行为学试验进一步证明, 2种组分比例的不同在花蓟马和西花蓟马种间识别过程中发挥重要作用。本研究推测花蓟马属的昆虫释放的聚集信息素组分可能是相同的。     

可用于黑刺粉虱快速鉴定的SCAR分子标记技术

针对粉虱类害虫难以准确快速地进行形态鉴别的问题, 以局部发生的黑刺粉虱Aleurocanthus spiniferus (Quaintance)为对象, 采用特征序列扩增区域 (SCAR) 标记法, 研究其快速分子检测技术。利用SCAR标记技术获得了长度为987 bp的黑刺粉虱特异性片段 (GenBank登录号为FJ613323), 根据此片段的碱基序列设计黑刺粉虱特异性引物1对(AS-F518/AS-R938), 其扩增片段为421 bp。种特异性检验结果显示, 该对引物只对黑刺粉虱的基因组具有扩增能力, 对同域发生的桔绿粉虱 Dialeurodes citri (Ashmead)以及其他种类的粉虱如烟粉虱Bemisia tabaci (Gennadius) B型、Q型、ZHJ-1型和ZHJ-2型, 温室粉虱Trialeurodes vaporariorum (Westwood)以及螺旋粉虱Aleurodicus disperses (Russell)等的基因组不具有扩增效果。该引物不仅对成虫具有良好的扩增效能, 对卵、2龄若虫和拟蛹等亦具有同样的扩增能力, 其最低检出限为1/1 920头成虫。该技术体系的建立在茶树和柑桔苗木调运的害虫检疫和监测/检测中具有重要意义。     

西花蓟马EST-SSR信息分析、标记筛选及其与Genomic-SSR的多态性比较

西花蓟马Frankliniella occidentalis是一种世界性入侵昆虫, 近年来传入我国并不断扩散蔓延。基于简单重复序列（simple sequence repeats, SSRs）的西花蓟马种群遗传结构研究对于揭示其传播途径等具有重要的指导价值。本研究对来源于西花蓟马的13 839条EST序列进行了uni-EST组装、 EST-SSR信息分析以及标记筛选, 并比较了EST-SSR与Genomic-SSR在分析遗传多样性方面的差异。结果表明： 在7 707个singlets中共找到2 623个SSR位点, 分布于1 930个uni-EST中, 平均每2.21 kb就出现一个SSR位点。重复单元中, 以单碱基重复单元为主（83.00%）, 其次是四碱基重复单元（11.17%）, 而二、 三、 五和六碱基重复单元所占比例较低（分别为1.41%, 0.80%, 2.02%和0.91%）。设计出的22对EST-SSR引物中, 4对引物能稳定扩增出清晰的目的条带; 荧光标记毛细管电泳发现3对引物表现出多态性。西花蓟马EST-SSR与Genomic-SSR多态性分析表明, 这3对多态性EST-SSR引物揭示的多态信息含量（PIC）为0.48~0.69, 比5对多态性Genomic-SSR引物揭示的PIC（0.88~0.92）略低。本研究结果可为今后更深入开展西花蓟马的种群遗传结构分析提供帮助。     

昆明地区菊苗上优势种害虫的发生规律

为明确菊苗上优势种害虫的发生规律及季节动态，采用五点取样法，利用粘虫板对2018—2020年温室种植的菊苗上优势种害虫进行调查，利用四分位法对优势种害虫发生期进行划分，同时分析了菊苗上优势种害虫的发生规律、季节动态及时间生态位。结果表明：西花蓟马在一年大部分时间均可危害菊苗，主要发生期集中在4—8月，3年的发生高峰分别在5月、6月和7月，此时西花蓟马的种群数量分别为636、1411和256头·板^-1,西花蓟马的生态位宽度呈现逐年降低的趋势；菊潜叶蝇的生态位宽度较小，每年的危害主要集中在1—6月，主要发生期在3—6月，3年的发生高峰均在3月，此时菊潜叶蝇的种群数量可达256、239和124头·板^-1;桃蚜和白粉虱在3年的生态位宽度较大，说明桃蚜和白粉虱在菊苗上的发生时间最长，主要发生期均在2—4月，发生高峰在全年内不固定。4种害虫间的生态位重叠指数相似，说明4种害虫在发生的时间维度上具有相似性且存在竞争关系。因此，在防治菊苗优势种害虫时应以西花蓟马和菊潜叶蝇为主，1—6月主要防治菊潜叶蝇，3—10月主要防治西花蓟马。     

Systemic effects of neem on western flower thrips,Frankliniella occidentalis (Thysanoptera: Thripidae)

The systemic effects of neem on the western flower thrips, Frankliniella occidentalis (Pergande), were investigated in laboratory trials using green bean, Phaseolus vulgaris L., in arena and microcosm experiments. In arena experiments, systemic effects of neem against western flower thrips larvae on primary bean leaves were observed with maximum corrected mortality of 50.6%. In microcosm experiments using bean seedlings, higher efficacy in the control of western flower thrips were observed with soil applications of neem on a substrate mixture (i.e., Fruhstorfer Erde, Type P, and sand) in a 1:1 ratio (93% corrected mortality) compared with application on the commercial substrate only (76% corrected mortality). However, longer persistence of neem was observed with soil application on the commercial substrate, which showed effects against thrips for up to 6 d after application. In addition to systemic effects observed on all foliage-feeding stages of western flower thrips, mortality on contact and repellent effects were observed on soil-inhabiting stages after soil applications of neem. Finally, bean seedlings grown from seeds pregerminated for 3 d in neem emulsion were also toxic to western flower thrips.     

Economic Thresholds of Western Flower Thrips (Thysanoptera: Thripidae) for Unripe Red Pepper in Greenhouse

This study was conducted to develop economic thresholds of western flower thrips (Frankliniella occidentalis Pergande) for unripe red pepper in greenhouses. To investigate the relationship between the density of thrips and resulting damages, experimental plots with five treatments (0, 4, 16, 48, 96 adults per plot) as initial release densities were established at the National Institute of Agricultural Science and Technology, Suwon, Korea, in 2004. Western flower thrips density was monitored using flower samplings and yellow sticky trap (8×13 cm) counts. Western flower thrips density was directly related to increased numbers of damaged fruits and reduced fruit yield. The number of marketable fruits produced decreased as the thrips densities increased. The major damage to pepper fruits caused by thrips was cosmetic scars that resulted from immature feeding. When flower samples or yellow trap caches were used to determine the density of thrips, which were collected on a previous sampling date, thrips densities were determined to be related to the percentage of fruits that were damaged, and a significant relationship was found for the flower samples (y = 0.3219x + 1.0792, r² = 0.8640 and for trap catches (y = 11.9209 log(x) −2.158, r² = 0.8306). The economically-tolerable ratio of damaged fruits based on control cost and market values under current greenhouse cultivation was estimated as 3.4 to 8.0%. Economic thresholds of western flower thrips ranged from 0.7 to 2.1 adults or nymphs per flower, and 2.3 to 5.7 adults per four-day sticky card count.     

Biotic resistance limits the invasiveness of the western flower thrips,Frankliniella occidentalis (Thysanoptera: Thripidae), in Florida

The spread of the western flower thrips, Frankliniella occidentalis (Pergande), has resulted in the world‐wide destabilization of established integrated pest management programs for many crops. It is hypothesized that frequent exposure to insecticides in intensive agriculture selected for resistant populations, which allowed invasive populations in the eastern USA to overcome biotic resistance from the native community of species. Research conducted in Florida to understand the role of biotic factors in limiting the abundance of the western flower thrips is reviewed. Orius spp. (Hemiptera: Anthocoridae) are effective predators that suppress populations of thrips on crop and non‐crop hosts in southern and northern Florida. Orius are more effective predators of the western flower thrips than the native flower thrips, F. tritici (Fitch) and F. bispinosa (Morgan). The native species are competitors of the western flower thrips. Excessive fertilization and the use of broad‐spectrum insecticides in crop fields further enhances populations of the western flower thrips. Interactions with native species clearly limit the abundance of western flower thrips in Florida, but populations are abundant in fertilized crop fields where application of insecticides excludes predators and competitor species.     

Spatial and temporal patterns of dispersal of western flower thrips (Thysanoptera: Thripidae) in nectarine orchards in British Columbia

Thrips were sampled from six nectarine orchards in the Dry Central Interior, British Columbia, Canada, between April and June 1993 using yellow sticky cards on posts spaced around the perimeter of each orchard. Although 12 identified species of thrips were captured, >90% of individuals were the western flower thrips, Frankliniella occidentalis (Pergande). The flight patterns and abundances of western flower thrips were compared between orchards located in two differently oriented valleys (N-S and E-W) and between orchards located close to or far from areas of wild land. Results indicate that densities of western flower thrips entering orchards, and their direction of movement, were related more to the external vegetation than either location within the two different valleys or general wind flow patterns. Western flower thrips tended to move into orchards close to ground level in early spring (late April and early May) but flew higher as ground cover grew taller and temperatures increased. Densities of western flower thrips at ground level were highest in an orchard with the densest dandelion ground cover. We conclude that the location of nectarine orchards in relation to wild areas is a major determinate of western flower thrips densities.     

Impact of production system on development of insecticide resistance in Frankliniella occidentalis (Thysanoptera: Thripidae)

The western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), has become one of the most difficult insects to control in the intensive agriculture of southeastern Spain. However, resistance problems are quite different in two neighboring areas, Murcia and Almeria, with distinct production systems. Thirty-six field populations of western flower thrips from sweet pepper crops were collected in two different dates in Murcia and Almeria in 2005 and 2006. Western flower thrips populations collected were exposed to a diagnostic concentration of spinosad, methiocarb, acrinathrin, and formetanate. The results allowed the recognition of higher levels of resistance in Almeria compared with Murcia throughout the growing season. The mortality at the diagnostic concentration for spinosad (120 ppm) in western flower thrips populations ranged from 34 to 81% in Almeria, and from 73 to 100% in Murcia. The mortalities at the diagnostic concentration to acrinathrin (800 ppm) and formetanate (8000 ppm) were 17-31% in Almeria and 77-100% in Murcia, and 14-41% in Almeria and 48-99% in Murcia, respectively, indicating large geographic variations. Toxicity of methiocarb was higher for western flower thrips populations from both areas. However, mortality at the diagnostic concentration of methiocarb (2000 ppm) varied from 56 to 90% in Almeria, and it was from 94 to 100% in Murcia. The impact of production systems and agricultural practices of each area on the development and stability of insecticide resistance is discussed.     

Effectiveness of imidacloprid (ProAgro 100 SL) in the control of glasshouse whitefly (Trialeurodes vaporariorum) and western flower thrips (Frankliniella occidentalis) on tomato under cover

In 2002 a glasshouse experiments were carried out on the effectiveness of ProAgro 100 SL in the control of glasshouse whitefly and western flower thrips on tomato cv. Perkoz. Strict observation of the basic prophylactic rules such as introducing in to glasshouse only healthy seedlings, uninfected by whitefly and thrips as well as isolation of tomato plants, especially the seedlings from ornamental plants are important to the limit of the pest population. During the vegetation period the population of whitefly and thrips may be limited by the insecticide application. The glasshouse experiments on the performance of ProAgro 100 SL in the control of mentioned above pest species were carried out at the Research Institute of Vegetable Crops in Skierniewice. The ProAgro 100 SL was applied in the concentration of 0,1%. The standard treatment was Juventox 040 SL (acetamiprid) in the concentration of 0.05%. The obtained results allow to find that ProAgro 100 SL in the rates of 0.1% performed well in the limiting population of whitefly and western flower thrips on glasshouse tomato.     

Management of thrips (Thysanoptera: Thripidae) in Australian strawberry crops: within-plant distribution characteristics and action thresholds

Abstract  Management of thrips in a hydroponic strawberry crop near Sydney, New South Wales, was studied from 1999 to 2002. The main species present were western flower thrips, Frankliniella occidentalis (Pergande), plague thrips, Thrips imaginis Bagnall, and, occasionally, onion thrips, Thrips tabaci Lindeman. Their within- and between-plant distribution and the degree and type of damage caused are described, and used to recommend action thresholds. Flowers are identified as the primary monitoring site, and young green fruit as the key damage site. Action thresholds of approximately 45% of flowers with 5 or more adult western flower thrips, or 40% of flowers with 10 or more adult plague thrips are recommended. Alternative thresholds using all motile stages are given for flowers, and for young green and red berries. A higher tolerance is recommended during cool, wet periods.     

Damage to nectarines by the western flower thrips (Thysanoptera: Thripidae) in the interior of British Columbia, Canada

The phenology of damage by the western flower thrips, Frankliniella occidentalis (Pergande), on nectarines was investigated using sticky cards and direct sampling of buds between 1993 and 1995 in the interior of British Columbia, the most susceptible period for damage by western flower thrips to nectarines. The life stage responsible for damage and variation in susceptibility to damage of 11 different nectarine varieties were determined. To evaluate the predictive ability of 2 sampling methods, thrips were counted from both buds and sticky cards before petal fall and correlated to damage levels at husk drop. Damage to nectarines was caused almost entirely by larval feeding at petal fall. No predictive relationships between adult or larval densities of western flower thrips and subsequent damage to fruit were apparent. Varieties did not differ in terms of larval densities at petal fall or the subsequent damage to fruit. Female western flower thrips oviposit in nectarine buds from dormant through bloom stages primarily in sepal tissues in the early buds, and in filaments and petals as these become available.     

Daily flight activity of the western flower thrips (Thysan., Thripidae) in nectarine orchards in British Columbia, Canada

Abstract:   Hourly flight patterns of western flower thrips, Frankliniella occidentalis (Pergande) (Thysan., Thripidae) were assessed over a 24 h period in two nectarine orchards located in the Similkameen Valley, British Columbia, on 13 separate occasions between March and October 1994. The numbers of western flower thrips (male or female, dark or pale morphs) trapped per hour at between four to eight heights (0.25–2.00 m) were compared with temperature, wind speed, time of year and location of trap placement. Traps were placed within orchards, at the border of orchards and adjacent wild land, and within wildlands. Temperature and wind speed thresholds are defined, although the effect of wind speed varied with height of flight. Height of flight was affected by both the height of prevalent vegetation and temperature, with mean height of flight increasing linearly with temperature. Numbers of thrips in flight decreased with height of sticky trap from the ground. The pale morph was generally more common than the dark morph of western flower thrips. Males were not present until after emergence of the first generation in May, and were found at an approximate ratio of 4 : 1 (F : M) for the rest of the year. There were no differences between sexes or morphs of western flower thrips for location, height or daily patterns of flight.     

Development of a novel bioassay for estimation of median lethal concentrations (LC50) and doses (LD50) of the entomopathogenic fungus Beauveria bassiana, against western flower thrips, Frankliniella occidentalis

To conduct laboratory experiments aimed at quantifying secondary acquisition of fungal conidia by western flower thrips (Frankliniella occidentalis), an efficient assay technique using Beauveria bassiana as the model fungus was developed. Various application protocols were tested and it was determined that the percent mortality did not vary among protocols. Peak mortality of second-instar nymphs, under constant exposure to conidia, occurred 5 days post-inoculation. Second-instar thrips that were exposed to conidia within 24 h of the molt to second instar were more susceptible to Beauveria bassiana than thrips exposed after times greater than 24 h post-molt. Conidia efficacy, which was monitored at 24 h intervals, did not differ significantly within 72 h. A test of the final bioassay system was conducted in a series of assays aimed at determining the LD50 of B. bassiana technical powder against second-instar western flower thrips. It was determined that B. bassiana (strain GHA) is highly effective at very low doses (LD50 of 33-66 conidia/insect).