Potential of a strain of the entomopathogenic fungus Beauveria bassiana (Hypocreales: Cordycipitaceae) as a biological control agent against western flower thrips,Frankliniella occidentalis (Thysanoptera: Thripidae)

Five Beauveria bassiana strains were evaluated for control of western flower thrips. Strain RSB was the most virulent, causing 69–96% mortality at concentrations of 1×10⁴–1×10⁷ conidia mL^?1, 10 days after inoculation of first instars. In greenhouse trials, RSB applied to broccoli foliage significantly reduced adult and larval populations.     

防治西花蓟马的病原真菌-白僵菌微菌核的液体培养

微菌核是许多植物病原真菌产生的越冬结构,目前还没有关于白僵菌微菌核的报道。用含不同氮源的介质培养出了2株白僵菌菌株,RSB和SZ21的微菌核。在液体培养条件下,2株菌株不仅能产生典型的芽孢和菌丝,还能产生微菌核。在以玉米粉(RSB和SZ21菌株分别为6.5×102和6.3×102mL-1)为氮源的介质中产生的微菌核浓度高于以大豆粉(RSB和SZ21菌株分别为2.5×102和2.2×102mL-1)为氮源的介质。菌株SZ21产生的芽孢浓度(在以大豆粉和玉米粉为氮源的介质中,分别为17.9×105和7.4×105L-1)高于RSB菌株的芽孢浓度(在以大豆粉和玉米粉为氮源的介质中,分别为14.8×105和6.2×105L-1)。含有硅藻土的微菌核制剂能在真空干燥(<5%湿度)后存活下来,并且对于活力没有显著影响。干燥后的微菌核颗粒在水洋菜培养基上复水和培养后,能够萌发菌丝和产生高浓度的分生孢子。将微菌核颗粒预混到土壤中能引起西花蓟马Frankliniella occidentalis(Pergande)土栖阶段的高死亡率。这是首次报道白僵菌的微菌核的培养,并为用虫生真菌防治土栖昆虫提供了一个新的方法。     

Potential use of the fungus Beauveria bassiana against the western flower thrips Frankliniella occidentalis without reducing the effectiveness of its natural predator Orius sauteri (Hemiptera: Anthocoridae)

Orius sauteri (Poppius; Hemiptera: Anthocoridae) is an important predator of western flower thrips, Frankliniella occidentalis (Pergande; Thysanoptera: Thripidae). O. sauteri would be directly exposed to the entomopathogenic fungus Beauveria bassiana (Bals.) Vuillemin in the field should the fungus be used as a biopesticide. If the fungus were to negatively affect O. sauteri in agro-ecosystems, predation of F. occidentalis by O. sauteri may be limited. The present study was undertaken to evaluate the insecticidal activity of strain B.bassiana-RSB of B. bassiana, which is highly virulent to F. occidentalis, on the predator under laboratory conditions. Results showed that, regardless of the concentration applied to first instars, Bb-RSB was not insecticidal against O. sauteri, nor did direct applications affect the developmental rate of the predator. Significant differences in developmental rates and adult longevity were observed between O. sauteri that fed on Bb-RSB-infected F. occidentalis cadavers and those that fed on untreated thrips. Developmental time (from first instar to adult) increased from 0.3 to 0.7 days and adult longevity decreased by 0.8 to 1.2 days for predators fed thrips treated with low and high concentrations of strain Bb-RSB, respectively, compared with predators fed on untreated thrips. However, these differences were only 3–13% of mean values for the controls, suggesting that the effects of Bb-RSB on O. sauteri are relatively minor. These findings highlight the potential use of O. sauteri in combination with B. bassiana for the biocontrol of F. occidentalis, but field tests must be performed to confirm their compatible use.     

Evaluation of the nematodes Steinernema feltiae and Thripinema nicklewoodi as biological control agents of western flower thrips Frankliniella occidentalis infesting chrysanthemum

In greenhouse studies, we evaluated a commercial formulation of the entomopathogenic nematode Steinernema feltiae and the inoculative release of the thrips-parasitic nematode Thripinema nicklewoodi against western flower thrips (WFT), Frankliniella occidentalis Pergande infesting potted chrysanthemums. Foliar sprays of S. feltiae applied at 1.25-2.5×10³ IJ mL^-1 and 1000 - 2000 L ha^-1 at 3-day intervals alone (targeting feeding stages) or in combination with soil applications (simultaneously treating non-feeding stages in the soil at the same rates) decreased but did not provide adequate control of thrips in flowering plants artificially infested with a dense population. Similar nematode treatments applied for four to five applications at 6-day intervals in two batches of initially clean chrysanthemums failed to prevent unacceptable damage to flowers and leaves from a dense natural infestation within the greenhouse. Although some IJ survived up to 48 h within flowers and flower buds, few nematode-infected thrips (larvae and adults) were recovered. In studies with T. nicklewoodi (which is not amenable for mass production), the inoculative releases of two parasitized hosts per plant enabled the nematode to become established within existing WFT populations under greenhouse conditions. However, relatively poor transmission and slow speed of kill (nematode primarily suppresses populations through host sterilization) prevented low level inoculations being effective over a single crop cycle. Further studies showed that transmission of T. nicklewoodi persisted for nine host generations, infected up to 83% of adult thrips and provided long-term suppression of discrete caged populations, but only after uneconomically high thrips densities had been reached.     

球孢白僵菌对黄瓜代谢产物及西花蓟马取食行为的影响

球孢白僵菌Beauveria bassiana灌根处理黄瓜后,发现西花蓟马Frankliniella occidentalis对叶片危害面积显著低于对照组;为探究产生这一现象的原因,测定黄瓜叶片中防御酶系和次生代谢产物等含量变化,采用直流刺吸电位仪(direct current electrical penetration graph,DC-EPG)分析西花蓟马成虫取食黄瓜叶片的行为.球孢白僵菌对黄瓜灌根后叶片中过氧化氢酶(CAT)、过氧化物酶(POD)和超氧化物歧化酶(SOD)含量均呈上升趋势,其中CAT活性增加最高,增加率为37.26％;丙二醛MDA的含量呈下降趋势;可溶性蛋白、类黄酮的含量增加.DC-EPG显示西花蓟马在经球孢白僵菌处理的黄瓜上,取食时间和次数均低于对照组.结果表明,黄瓜植株经球孢白僵菌处理后防御酶物质增加,进而导致西花蓟马对叶片危害下降,利于对西花蓟马的防治.     

西花蓟马的鉴别及其与近缘种的区别

西花蓟马是我国2 0 0 3年在北京新发现的外来入侵害虫,因虫体很小鉴定困难。该文提供了西花蓟马详细的形态鉴别特征,同时给出了与西花蓟马相似的烟蓟马、花蓟马和佛罗里达花蓟马共4种花蓟马的鉴定检索表。     

Engineered multidomain cysteine protease inhibitors yield resistance against western flower thrips (Frankliniella occidentalis) in greenhouse trials

Western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), cause very large economic damage on a variety of field and greenhouse crops. In this study, plant resistance against thrips was introduced into transgenic potato plants through the expression of novel, custom-made, multidomain protease inhibitors. Representative classes of inhibitors of cysteine and aspartic proteases [kininogen domain 3 (K), stefin A (A), cystatin C (C), potato cystatin (P) and equistatin (EIM)] were fused into reading frames consisting of four (K-A-C-P) to five (EIM-K-A-C-P) proteins, and were shown to fold into functional inhibitors in the yeast Pichia pastoris. The multidomain proteins were expressed in potato and found to be more resistant to degradation by plant proteases than the individual domains. In a time span of 14-16 days, transgenic potato plants expressing EIMKACP and KACP at a similar concentration reduced the number of larvae and adults to less than 20% of the control. Leaf damage on protected plants was minimal. Engineered multidomain cysteine protease inhibitors thus provide a novel way of controlling western flower thrips in greenhouse and field crops, and open up possibilities for novel insect resistance applications in transgenic crops.     

Increased damage by western flower thrips Frankliniella occidentalis in chrysanthemum intercropped with subterranean clover

Suppressive effects of intercropping on Frankliniella occidentalis (Pergande) infestations have been reported in several crops. However, this study demonstrates that in year-round chrysanthemum, Dendranthema grandiflora Tzvelev, undersowing with subterranean clover, Trifolium subterraneum L., results in an increased thrips feeding damage. In a pot experiment, performed with chrysanthemum plants (cultivars Reagan and Tiger) grown with or without subterranean clover, significantly more leaves with silver and growth damage were found in the chrysanthemum plants with subterranean clover in comparison with the monocropped chrysanthemum plants. Similarly, the degree of deformation of leaf perimeter and leaf surface was higher in the top leaves of the intercropped chrysanthemum plant. In the soil experiment (only performed with Tiger and plants were grown in the soil in the greenhouse) intercropped chrysanthemum plants suffered a higher feeding damage as well. Analysis of the relation between silver or growth damage and the thrips pressure demonstrates that at similar thrips pressure in the intercropped chrysanthemum plants suffered significantly more damage. The higher thrips pressure in the intercropped chrysanthemum only explains the differences in damage partly. Changes in the reaction of chrysanthemum plants to thrips feeding or in the behaviour of the thrips, mediated by the presence of the non-host crop, are discussed. Our explanation is that chrysanthemum plants grown with clover are more susceptible to thrips feeding than monocropped plants. We conclude that undersowing with clover does not contribute to reduce damage by F. occidentalis in year-round chrysanthemum. Also, the influence of crop diversification on a pest cannot be foreseen until the specific characteristics of each individual crop – pest system are studied.     

A tritrophic effect of host plant on susceptibility of western flower thrips to the entomopathogenic fungus Beauveria bassiana

Adult female western flower thrips (Frankliniella occidentalis) were exposed 12-24h to bean (Phaseolus vulgaris) and impatiens (Impatiens wallerana) leaf disks treated with Beauveria bassiana conidia and then transferred to clean bean or impatiens at various times post-treatment. Significantly greater levels of fungal infection were observed when thrips were treated on bean versus impatiens, but exposure to impatiens following treatment had no effect on fungal infection (percent mortality). This result, combined with observations of no inhibition of germination of conidia exposed to intact or macerated impatiens foliage, indicated that the negative effect of the impatiens host plant was not due to plant chemical compounds (antibiosis). Further observations revealed that insects acquired (picked-up) 75% more conidia from treated bean disks than from treated impatiens disks. This difference in dose acquisition was determined to account for the observed difference in percent mortality (15%) following treatment on the two host plants. Median lethal doses (LD(50)) of B. bassiana were not significantly different on the two host plants, but median lethal concentrations were nearly 7-fold greater on impatiens. This difference was explained by disproportionate rates of conidial acquisition at measured rates of conidial deposition (an inverse relationship was observed between application rate expressed as conidia/mm(2) and the number of conidia acquired). The mechanism underlying the differential rates of conidial acquisition from bean versus impatiens was not determined.     

The spread of the western flower thrips Frankliniella occidentalis (Pergande)

Abstract 1 Since the late 1970s, the western flower thrips has spread from its original distribution in western North America to become a major worldwide crop pest. 2 A wide range of data sources have been used to map the original distribution in the U.S.A. and Canada, and the progress of the spread in the U.S.A., Canada, Europe, northern Africa and Australia. 3 The possible reasons for the start of the spread are discussed. The most likely reason is that intensive insecticide use in horticulture in the 1970s and 1980s selected an insecticide resistant strain or strains. These then established in glasshouses across North America and spread from there to Europe, Asia, Africa and Australia. 4 The international spread of the western flower thrips occurred predominantly by the movement of horticultural material, such as cuttings, seedlings and potted plants. Within Europe, an outward spread from the original outbreak in the Netherlands is discernible. The speed of spread was 229 ± 20 km/year. 5 The spread has not been restricted to glasshouses. The western flower thrips has established outdoors in areas with milder winters; for example, across the southern U.S.A., southern Europe and Australia. It also overwinters in some regions with colder winters. 6 Polyphagous phytophagous thrips have many factors predisposing them to become worldwide crop pests, particularly in glasshouses. Some other species that might spread in a similar way to the western flower thrips are listed.     

利用冷冻切片对球孢白僵菌侵染西花蓟马过程的观察

为探索球孢白僵菌对西花蓟马的侵染,研究球孢白僵菌在西花蓟马体表和虫体内的侵染过程,采用冷冻切片与HE染色技术,对被球孢白僵菌侵染的西花蓟马成虫进行组织切片及HE染色。结果表明,接菌4 h,体表粘附的孢子开始萌发,8-12 h芽管继续伸长并出现穿透体壁。18 h虫菌体出现在西花蓟马体内,球孢白僵菌成功侵入西花蓟马体内。之后,西花蓟马体内虫菌体、菌丝明显增多,各组织器官均受到感染,消化道、肌肉组织等发生病变。     

Selection of promising fungal biological control agent of the western flower thrips Frankliniella occidentalis (Pergande)

Aims: Larval stages of Frankliniella occidentalis are known to be refractory to fungal infection compared with the adult stage. The objective of this study was to identify promising fungal isolate(s) for the control of larval stages of F. occidentalis. Methods and Results: Ten isolates of Metarhizium anisopliae and eight of Beauveria bassiana were screened for virulence against second‐instar larvae of F. occidentalis. Conidial production and genetic polymorphism were also investigated. Metarhizium anisopliae isolates ICIPE 7, ICIPE 20, ICIPE 69 and ICIPE 665 had the shortest LT₅₀ values of 8·0–8·9 days. ICIPE 69, ICIPE 7 and ICIPE 20 had the lowest LC₅₀ values of 1·1 × 10⁷, 2·0 × 10⁷ and 3·0 × 10⁷ conidia ml^?1, respectively. Metarhizium anisopliae isolate ICIPE 69 produced significantly more conidia than M. anisopliae isolates ICIPE 7 and ICIPE 20. Internally transcribed spacers sequences alignment showed differences in nucleotides composition, which can partly explain differences in virulence. Conclusion: These results coupled with the previous ones on virulence and field efficacy against other species of thrips make M. anisopliae isolate ICIPE 69 a good candidate. Significance and Impact of the Study: Metarhizium anisopliae isolate ICIPE 69 can be suggested for development as fungus‐based biopesticide for thrips management.     

Effects of post-application irrigation and substrate moisture on the efficacy of entomopathogenic nematodes against western flower thrips, Frankliniella occidentalis

The efficacy of entomomatogenic nematodes (Steinernema bicornutum Tallosi, Peters and Ehlers and/or Heterorhabditis indica LN2 Poinar, Karunakar and David) against the soil‐dwelling life stages of western flower thrips Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) was assessed under different moisture conditions in a commercial plant‐growing substrate in laboratory experiments. In the first experiment, both nematode species were tested at substrate moisture ranges of 67, 78, 88, or 95% relative moisture content, that were maintained before applying the nematodes at 100 or 400 infective juveniles (IJs) cm^?2. In the second experiment, 10, 25, 50, 100, or 120 ml irrigation water, resulting in relative moisture contents of 72, 81, 90, 99%, or more than the saturation level of the substrate, respectively, was applied to the substrate. Heterorhabditis indica LN2 was applied either in 3 ml water and followed by irrigation, or by suspending the infective juveniles in the water amounts indicated above to apply the nematodes in higher volume. Results indicated that at the higher application rate, initial moisture content did not significantly affect the efficacy of H. indica LN2. On the other hand, increasing moisture content resulted in an improved efficacy of H. indica LN2 and S. bicornutum at lower and higher application rates, respectively. Similar thrips control levels of 44 and 60% at the lower and higher application rate of H. indica LN2, respectively, were obtained at 88% relative moisture content. In the second experiment, higher and statistically similar thrips mortality of 40 and 50% at lower and higher application rates of H. indica LN2, respectively, were obtained when the infective juveniles were applied in a high volume suspension of 100 ml, or when followed by irrigation with 25 ml water, resulting in both cases in 81% relative moisture content. Generally, efficacies of the nematodes for thrips control can be improved by using an appropriate moisture content and/or post‐application irrigation. Thus, the high nematode application rates required for successful F. occidentalis control can be partly attributed to substrate moisture content and/or post‐application irrigation.     

球孢白僵菌不同分离株对西花蓟马的毒力测定

【背景】西花蓟马是我国重要的一种入侵性害虫,已对多种化学农药产生了抗性。【方法】采用孢子悬浮液喷雾处理生测法,研究了8个球孢白僵菌菌株BbKM030716、BbCG051229、BbJS080625、BbQJ031121、BbXW060615、BbSM090521、BbYY090613和BbMZ051230对西花蓟马成虫的毒力,并用＂时间—剂量—死亡率＂模型进行了分析。【结果】供试球孢白僵菌各菌株对西花蓟马成虫侵染致病的剂量效应参数值分别为1.43、0.87、0.93、0.98、1.23、0.92、1.07和0.86。用1.25×104～1.25×108个·mL-1孢子悬浮液接种后,连续10d内西花蓟马的校正死亡率分别为44.13%～98.49%、12.63%～78.90%、30.36%～96.92%、51.36%～98.74%、26.14%～98.59%、7.27%～78.71%、49.06%～98.74%和27.67%～87.36%。球孢白僵菌对西花蓟马成虫的致死剂量是时间的函数,各菌株对西花蓟马成虫的致死中时随孢子浓度的增大而逐渐减小。【结论与意义】球孢白僵菌菌株BbKM030716、BbJS080625、BbQJ031121和BbXW060615对西花蓟马成虫具有较强毒力,可作为西花蓟马生防制剂开发的潜力菌株。     

Dehydration yields distinct transcriptional shifts associated with glycogen metabolism and increases feeding in the western flower thrips,Frankliniella occidentalis

We examined water balance characteristics and the influence of desiccating conditions on the physiology and behavior of adult western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). Western flower thrips are globally invasive and likely to contend with shifts in water availability across their expansive geographic range. Basic water balance characteristics, including water mass and dry mass, were established for adult males and females, revealing a distinct sexual dimorphism wherein females are larger, and males retain a larger percentage of their body mass as water. Males lose relative water mass more quickly and their survival times are shorter compared to females. RNA-seq analysis identified significant enrichment of a multitude of factors including carbohydrate transport and metabolism in dehydrated males and females. This was validated by altered glycogen levels, suggesting a rapid depletion in glycogen during dehydration. The probability of thrips feeding significantly increases when desiccation occurs, potentially to replenish water content and nutrient reserves. Our results establish the fundamental water balance characteristics of adult thrips and indicate that dehydration significantly influences the survivorship and feeding behavior of thrips; all of which being crucial factors that contribute to their capacity as vectors for plant pathogens.     

Optimization of dietary RNA interference delivery to western flower thrips Frankliniella occidentalis and onion thrips Thrips tabaci

In insect reverse genetics, dietary delivery of interfering RNAs is a practical approach in nonmodel species, such as thrips, whose small size, and feeding behavior restricts the use of other delivery methods. In a laboratory context, an unsuitable diet could confound the interpretation of an RNA interference (RNAi) phenotype, however well-formulated artificial diets can minimize experimental variability, reduce the need for insect handling, and can further be used for roles, such as delivering double-strand RNA (dsRNA)-expressing recombinant bacteria. In this study, artificial diets for oral delivery of dsRNA were developed for two important pest thrips species, western flower thrips (Frankliniella occidentalis) and onion thrips (Thrips tabaci), with the goal of (a) stimulating feeding behavior, (b) supporting optimal growth rates of dsRNA-expressing symbiotic bacteria, and (c) nutritionally supporting the thrips for sufficient periods to observe RNAi phenotypes. The efficacy of artificial diets for ingesting “naked” dsRNA or dsRNA-expressing symbionts and dsRNA delivery via host plant uptake was evaluated. Compared with previously published diet formulations, new combinations based on tryptone, yeast, and soy were superior for enhancing feeding and longevity. However, simply adding “naked” dsRNA to an artificial diet was an unreliable form of RNAi delivery in our hands due to dsRNA degradation. Delivery via host plants was more successful, and the new diet formulation was suitable for symbiont-mediated dsRNA delivery, which we believe is the most convenient approach for large-scale knockdown experiments. This study, therefore, provides alternative methodologies for thrips rearing, dietary RNAi delivery, and insights into the challenges of performing dietary RNAi in nonmodel insects.     

Acquisition of lethal doses of Beauveria bassiana conidia by western flower thrips, Frankliniella occidentalis, exposed to foliar spray residues of formulated and unformulated conidia

Secondary acquisition of Beauveria bassiana conidia was recorded on the whole bodies and selected body parts of second-instar nymphs and adult female western flower thrips exposed to foliar spray residues of three differently formulated conidial preparations, for 24 h. Conidia were formulated in emulsifiable oil or with clay (wettable powder), or were essentially unformulated conidia (technical grade powder suspended in water with a surfactant). Formulation had no significant effect on dose acquisition and no effect on virulence of acquired conidia. The mean nymphal LC50/LD50 was 116 conidia/mm2 and 52conidia/insect, respectively; the values for adults were 19 conidia/mm2 and 5conidia/insect. Greatest numbers of conidia were recorded on the legs and abdomens of nymphs and on the legs, wings, and thoraces of adults. As would be expected, numbers of conidia acquired increased with residue concentration (application rate). However, an inverse relationship was noted between acquisition rate (conidia acquired/total conidia applied) and residue concentration. The mechanism underlying this response was not determined. However, there was no indication that any body parts (e.g., tarsi) became saturated with spores, which suggests that either the thrips were repelled by the conidial residues or that as the concentrations of conidia on the substrate increased, conidia somehow became more difficult to acquire. Slopes of the LC probit regressions were lower than those of the LD regressions (mean 1.14 vs 1.78), suggesting that the low slopes often obtained in fungal pathogen assays could be partly an artifact of unequal rates of dose acquisition at low vs high application rates.     

低温胁迫对西花蓟马抗氧化酶活性的影响

西花蓟马Frankliniella occidentalis(Pergande)是一种重要入侵害虫。温度是影响西花蓟马生长发育和繁殖的一个重要非生物因子,西花蓟马对温度的耐受性决定了它的越冬存活率和地理分布。本文研究了4种抗氧化酶在低温胁迫下的活力变化。结果表明,4种抗氧化酶对低温胁迫具有不同的响应模式。低温处理能显著提高SOD的活力,对POD的活力则具有明显的抑制作用;然而,CAT和GST的活力在低温胁迫处理表现为先降低,随后逐渐提高的趋势。本研究结果表明4种抗氧化酶在西花蓟马应对低温胁迫中具有重要作用。     

不同温度下双尾新小绥螨对西花蓟马的捕食功能反应

为明确双尾新小绥螨Neoseiulus bicaudus各螨态对西花蓟马Frankliniella occidentalis不同龄期若虫的捕食量及不同温度对其捕食量的影响。本研究通过测定不同温度下24 h内双尾新小绥螨各螨态对不同密度西花蓟马若虫的捕食情况(西花蓟马猎物密度分别为5、10、15、20和25头/室,温度分别设置为23、26、29、32和35℃)。研究结果表明,双尾新小绥螨各螨态对猎物的捕食量随猎物密度的升高而增加。当温度为29℃,雌成螨对西花蓟马1龄若虫捕食量最高,在猎物数为25头/室时,可达4.4头/日。23~32℃内,双尾新小绥螨的雌成螨的捕食量随温度的上升而增加,到达35℃时开始降低。29~32℃双尾新小绥螨的雌成螨对西花蓟马1龄若虫的捕食量最高,可达6.7头/日,最适于其捕食。结果表明双尾新小绥螨对西花蓟马若虫的捕食功能反应类型符合Holling II模型,其对西花蓟马有较好的捕食能力,可利用于西花蓟马的生物防治。