昆虫病原线虫对腰果细蛾的致病力测定

室内测定了Steinernema carpocapsae All、S.carpocapsae Biosys、S.glaseri NC52、S.longicaudum X-7、Heterorhabditis bacteriophora H06和H.indica LN2共6个昆虫病原线虫品系对腰果细蛾Acrocercops syngramma Meyrick幼虫的致病力。结果表明,线虫S.carpocapsae All、Biosys品系与线虫S.glaseri NC52对腰果细蛾幼虫的致死效果显著高于其它线虫,但供试的6个线虫品系均未能在腰果细蛾幼虫体内繁殖下一代,说明供试线虫在腰果细蛾幼虫体内难以繁殖。选择小卷蛾斯氏线虫S.carpocapsae All进一步研究其对腰果细蛾的致病力,结果发现,All线虫对腰果细蛾3龄幼虫处理24 h后的致死中浓度LC50为19.88 IJs/虫。腰果细蛾各龄幼虫和蛹对小卷蛾斯氏线虫S.carpocapsae All均表现出高度的敏感性,但不同幼虫龄期和处理时间敏感程度不同。小卷蛾斯氏线虫S.carpocapsae All对不同龄期腰果细蛾幼虫和蛹均表现出较强的致病力。     

4种昆虫病原线虫对草地贪夜蛾的致死作用

【目的】探讨昆虫病原线虫对草地贪夜蛾幼虫的侵染效果。【方法】在室内条件下采用生测试验法,测定了小卷蛾斯氏线虫AⅡ、长尾斯氏线虫X-7、夜蛾斯氏线虫SN和嗜菌异小杆线虫H06等4种昆虫病原线虫对草地贪夜蛾2龄和5龄幼虫的致死作用。【结果】小卷蛾斯氏线虫AⅡ与草地贪夜蛾数量比为50∶1时,36 h后草地贪夜蛾2龄和5龄幼虫的死亡率分别为92%和100%。长尾斯氏线虫X-7与草地贪夜蛾数量比为50∶1时,36 h后草地贪夜蛾2龄幼虫的死亡率为80%;在数量比30∶1时,36 h后草地贪夜蛾5龄幼虫死亡率为100%。夜蛾斯氏线虫SN与草地贪夜蛾数量比为400∶1时,36 h后草地贪夜蛾2龄和5龄幼虫的死亡率分别为88%和85%。嗜菌异小杆线虫H06与草地贪夜蛾在400∶1时,36 h后草地贪夜蛾2龄和5龄幼虫的死亡率分别为32%和67.5%。【结论】小卷蛾斯氏线虫AⅡ具有较好的草地贪夜蛾生物防治潜质,其次是长尾斯氏线虫X-7。     

病原线虫对桔小实蝇种群的控制作用

通过室内和田间实验研究了昆虫病原线虫对桔小实蝇Bactrocera (Bactrocera) dorsalis (Hendel)的控制作用。室内实验结果表明,供试的3种线虫的4个品系（小卷蛾斯氏线虫Steinernema carpocapsae All品系与A24品系,夜蛾斯氏线虫Steinernema feltiae SN品系和嗜菌异小杆线虫Heterorhabditis bacteriophora H06品系）,以小卷蛾斯氏线虫All品系对桔小实蝇的侵染力最强,其3天的LD₅₀和LD₉₅分别为35.0和257.1条/cm²土壤。按300条/cm²土壤的量施用,小卷蛾斯氏线虫All品系对当代桔小实蝇的控制效果为86.3%。用以虫期作用因子组建的生命表方法评价了小卷蛾斯氏线虫All品系对田间桔小实蝇下代种群的控制作用,结果表明,按300条/cm²土壤的量施用线虫,对照杨桃园的桔小实蝇种群趋势指数为105.9,而处理杨桃园的桔小实蝇种群趋势指数下降为15.5;小卷蛾斯氏线虫All品系对桔小实蝇的干扰控制指数为0.146,即线虫处理果园的下代种群密度仅为对照果园的14.6%。     

噻虫嗪对昆虫病原线虫侵染韭菜迟眼蕈蚊能力的影响

【目的】为提高昆虫病原线虫对韭菜迟眼蕈蚊Bradysia odoriphaga Yang et Zhang幼虫的防治效果,将昆虫病原线虫与环境友好型化学杀虫剂混用是一条有效途径。【方法】测定了噻虫嗪与6个昆虫病原线虫品系混用对韭菜迟眼蕈蚊的作用效果,以及温度和土壤含水量对作用效果的影响,并进行了田间验证。【结果】田间推荐浓度噻虫嗪(100 mg·L~(-1))对6种供试线虫存活无显著影响。处理后3 d,低浓度噻虫嗪(15 mg·L~(-1))分别与6品系线虫混合后处理韭菜迟眼蕈蚊幼虫,其死亡率明显高于线虫和噻虫嗪单用处理。小卷蛾斯氏线虫SF-SN+噻虫嗪、印度异小杆线虫LN2+噻虫嗪和小卷蛾斯氏线虫All+噻虫嗪3种组合发挥杀虫作用的最适温度范围分别为20~25℃、25~30℃和25~30℃,显著高于其他温度;最适土壤含水量范围为10%~18%,也显著高于其他湿度。大田条件下,施用后7 d,单用噻虫嗪、线虫+噻虫嗪组合处理对韭菜迟眼蕈蚊的防治效果显著高于单线虫,且以芫菁夜蛾斯氏线虫SF-SN+噻虫嗪的防治效果最高,达到93%以上。【结论】芫菁夜蛾斯氏线虫SF-SN品系与噻虫嗪组合联合防治韭菜迟眼蕈蚊效果最好。     

芜菁夜蛾线虫和嗜菌异小杆线虫寄生特性的比较

对2种昆虫病原线虫(芫菁夜蛾线虫Steinernema feltiae Filipjev和嗜菌异小杆线虫Heterorhaditis bacteriphora Poinar)的生物学特性进行研究。结果表明:在25℃下,在大蜡螟Galleria mellonella L.幼虫体内完成1个侵染周期芫菁夜蛾线虫Otio品系需220～230h,嗜菌异小杆线虫15-2品系需200～210h。15-2品系比Otio品系具有较强的繁殖力。2种线虫侵染大蜡螟幼虫的温度范围均为20～30℃,在此温度范围内,Otio品系的侵染力和运动能力均强于15-2品系。当土壤含水量(w/w)为5%～15%,线虫的侵染活性最高,随湿度降低线虫的侵染活性明显降低。     

小卷蛾斯氏线虫侵染对草地贪夜蛾幼虫天然免疫反应的影响

【目的】探讨昆虫病原线虫小卷蛾斯氏线虫Steinernema carpocapsae All侵染对草地贪夜蛾Spodoptera frugiperda幼虫天然免疫反应的影响。【方法】借助倒置显微镜观察和鉴定草地贪夜蛾幼虫的血细胞类型,并对小卷蛾斯氏线虫侵染后不同时间的草地贪夜蛾幼虫血细胞总数目进行统计;通过倒置显微镜观察草地贪夜蛾幼虫对侵入的小卷蛾斯氏线虫的包囊反应;利用倒置荧光显微镜观察小卷蛾斯氏线虫侵染后的草地贪夜蛾幼虫血细胞对金黄色葡萄球菌Staphylococcus aureus的吞噬活性;检测小卷蛾斯氏线虫侵染后的草地贪夜蛾幼虫血淋巴中酚氧化酶(phenoloxidase, PO)活性、体内抗菌肽基因相对表达水平以及血浆的抗菌活性。【结果】从草地贪夜蛾幼虫体内共发现5种不同类型的血细胞,分别为原血细胞、粒细胞、类绛色细胞、珠血细胞和浆血细胞。注射1 μL侵染期(infective juveniles, IJs)小卷蛾斯氏线虫(3 IJs/μL)后9和12 h,草地贪夜蛾幼虫的血细胞总数目显著增多。草地贪夜蛾幼虫的血细胞不能包囊活的以及冷处死的小卷蛾斯氏线虫,但可以包囊热处死的线虫。活的小卷蛾斯氏线虫会显著抑制草地贪夜蛾幼虫血细胞对金黄色葡萄球菌的吞噬活性,但冷处死和热处死的线虫不能。注射1 μL(3 IJs/μL)小卷蛾斯氏线虫后,草地贪夜蛾幼虫血淋巴PO活性总体呈“下降 升高 下降”变化趋势;体内抗菌肽基因Attacin-A2, Attacin-B1, Cecropin-B3, Cecropin-D, Gallerimycin, Gloverin-3以及Lebocin-2的表达水平在线虫侵染后12 h时显著上调,24 h时恢复到对照水平或低于对照水平;血淋巴抗菌活性水平在小卷蛾斯氏线虫侵染后12 h时显著升高,24 h时与对照无显著差异。【结论】小卷蛾斯氏线虫在侵入早期会抑制草地贪夜蛾幼虫的天然免疫反应来建立感染;随后草地贪夜蛾的免疫系统会被激活试图抵御小卷蛾斯氏线虫的侵染;后期随着线虫的成功定殖,草地贪夜蛾的免疫系统最终被抑制或破坏。本研究所得结果为进一步揭示线虫 草地贪夜蛾的免疫互作机理奠定了基础,也为改善昆虫病原线虫对草地贪夜蛾的防治效果提供了理论依据。     

不同昆虫病原线虫一共生细菌组合的生化特性研究

本文分别以无菌的小卷蛾斯氏线虫的A24品系线虫与其自身携带的共生细菌菌株和与格氏线虫RS92品系的共生细菌菌株,以及以无菌的嗜菌异小杆线虫的H06品系线虫与其自身携带的共生细菌菌株和与大异小杆线虫HNA品系的共生细菌菌株建立单菌组合;测定了各组合感染期线虫的致病力、干重和体内主要生化物质的含量.结果发现,共生细菌除了对感染期线虫的致病力产生显著影响外,对线虫干重、蛋白质、氨基酸、糖原和脂肪酸等生化物质的含量也有很大的影响.     

昆虫病原线虫对黄地老虎致病力的研究

为了探索应用昆虫病原线虫防治黄地老虎Agrotis segetum的可行性,室内生测比较了9个昆虫病原线虫品系对黄地老虎4龄幼虫的致病力。生测结果表明,9个不同昆虫病原线虫品系对黄地老虎4龄幼虫的致病力差别很大,其中格氏线虫Steinernema glaseri ib(Sgib)是对黄地老虎幼虫致病力最强的昆虫病原线虫品系,侵染24、36和72 h黄地老虎幼虫的校正死亡率均高于其他线虫品系。此外,还测定了不同温度、土壤湿度以及线虫剂量对格氏线虫Sgib致病力的影响,结果表明,在21℃-27℃、20%土壤湿度以及侵染期线虫与黄地老虎幼虫数量比不少于10∶1的条件下,格氏线虫Sgib对黄地老虎幼虫的致病效果最好。田间小区实验表明,格氏线虫Sgib对黄地老虎幼虫的防效达到96.67%。     

不同品系昆虫病原线虫对烟草小地老虎的致病力

小地老虎Agrotis ypsilon Rottemberg是烟田的一种重要的地下害虫。为了探明不同品系的昆虫线虫对烟田地下害虫小地老虎的致病力,在室内采用生测试验法,以滤纸为介质,测定了斯氏属线虫Steinernema carpocapsae和异小杆属线虫Heterorhabditis bacteriophora共5个品系对小地老虎3龄幼虫、5龄幼虫和蛹的致病力。结果表明,5个不同昆虫病原线虫品系均对小地老虎3龄幼虫具有致病力,其中斯氏线虫S.carpocapsae-A11(Sc-A11)对小地老虎的致死速度最快,侵染24 h、36 h和72 h后小地老虎校正死亡率分别为41.3%、69.7%和86.7%,均高于其它线虫品系,小地老虎的死亡率随着线虫侵染时间的增加而增加的,至96 h后校正死亡率达到100%,Sc-A11侵染5龄幼虫的校正死亡率均高于其它线虫品系,可见Sc-A11对小地老虎幼虫的致病力最强,为最敏感的昆虫病原线虫品系。Sc-All对小地老虎3龄幼虫、5龄幼虫的校正死亡率在72 h时分别为86.7%、43.3%,化蛹率为23.7%,Sc-All不侵染小地老虎蛹。盆栽烟草上线虫Sc-A11剂量为20头IJs/cm~2施用3次和线虫剂量为80头IJs/cm~2施用2次的处理的保苗效果最好,3 d后保苗效果分别为79.64%和76.85%,达到最佳的防治效果。     

昆虫病原线虫对草原毛虫幼虫的致病力

草原毛虫Gynaephora alpherakii是青藏高原草原的主要害虫。国家明令禁止使用化学药剂防控这一害虫。因此,有必要以生物农药控制这种害虫。本试验室内测定5种昆虫病原线虫（Steinernema longicaudum X-7,S. carpocapsae All,S. feltiae SN,Heterorhabditis bacteriophora H06和H. indica LN2）对草原毛虫的致病力。结果表明,S. carpocapsae All、S. feltiae SN两种线虫对草原毛虫的侵染效果较好。适宜S. carpocapsae All、S. feltiae SN线虫的侵染活动的环境温度为25℃;低温环境下两种线虫随着剂量的提高,对草原毛虫的致病力均有着显著地增长（P<0.05）,且S. feltiae SN线虫的活性高于S. carpocapsae All线虫。因此,昆虫病原线虫S. feltiae SN品系具有效防治青藏高原高寒草地草原毛虫的应用潜力。     

Distribution patterns of entomopathogenic nematodes applied through drip irrigation systems

The distribution of entomopathogenic nematodes applied by drip irrigation was evaluated by injecting small volumes of Steinernema carpocapsae (Weiser) All strain, Steinernema feltiae (Filipjev) SN strain, Steinernema glaseri Steiner, and Heterorhabditis bacteriophora HP 88 strain Poinar suspensions into drip irrigation lines. Additionally, Steinernema riobrave Cabanillas, Poinar, & Raulston, and S. carpocapsae were injected in a 10-liter volume of water with an injection pump. Overall, the nematodes were evenly distributed along the drip lines. The total number of nematodes recovered from drip emitters was variable ranging from 42 to 92%. However, drip irrigation lines have potential to deliver entomopathogenic nematodes efficiently into pest habitats.     

Virulence of entomopathogenic nematodes to pecan weevil larvae, Curculio caryae (Coleoptera: Curculionidae), in the laboratory

The pecan weevil, Curculio caryae (Horn), is a key pest of pecans in the Southeast. Entomopathogenic nematodes have been shown to be pathogenic toward the larval stage of this pest. Before this research, only three species of nematodes had been tested against pecan weevil larvae. In this study, the virulence of the following nine species and 15 strains of nematodes toward fourth-instar pecan weevil was tested: Heterorhabditis bacteriophora Poinar (Baine, HP88, Oswego, NJ1, and Tf strains), H. indica Poinar, Karunakar & David (original and Homl strains), H. marelatus Liu & Berry (IN and Point Reyes strains), H. megidis Poinar, Jackson & Klein (UK211 strain), H. zealandica Poinar (NZH3 strain), Steinernema riobrave Cabanillas, Poinar & Raulston (355 strain), S. carpocapsae (Weiser) (All strain), S. feltiae (Filipjev) (SN strain), and S. glaseri (Steiner) (NJ43 strain). No significant difference in virulence was detected among nematode species or strains. Nematode-induced mortality was not significantly greater than control mortality (in any of the experiments conducted) for the following nematodes: H. bacteriophora (Baine), H. zealandica (NZH3), S. carpocapsae (All), S. feltiae (SN), S. glaseri (NJ43), and S. riobrave (355). All other nematodes caused greater mortality than the control in at least one experiment. Heterorhabditis megidis (UK211) but not H. indica (original) displayed a positive linear relationship between nematode concentration and larval mortality. Results suggested that, as pecan weevil larvae age, they may have become more resistant to infection with entomopathogenic nematodes.     

Distribution of entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) in natural habitats in California, USA

A total of 270 soil samples from 30 different habitats in 10 geographic regions of California were evaluated for the presence of rhabditid entomopathogenic nematodes. Nematodes were isolated from 26.3% of the samples. The recovered isolates were identified as Steinernema carpocapsae, S. feltiae, S. kraussei, S. longicaudum, S. oregonense, Heterorhabditis marelatus and H.bacteriophora. Among the steinernematids, S. kraussei and S. feltiae were the most commonly encountered species, generally occurring in acidic soils high in organic matter. Among the heterorhabditids, H. bacteriophora was isolated along the southern coast, whereas H. marelatus was recovered along the northern coast of California. Steinernematids were recovered from coniferous forests, oak woodlands and grasslands whereas heterorhabditids were isolated from coastal marshes.     

EVect of the Entomopathogenic Nematodes, Steinernema feltiae and S. carpocapsae , on the Potato Cyst Nematode, Globodera rostochiensis , in Pot Trials

Two pot experiments, one in a glasshouse and the other in an outdoor sand plunge, were conducted to examine the influence of the entomopathogenic nematodes, Steinernema feltiae and S. carpocapsae , on the invasion and development of the potato cyst nematode, Globodera rostochiensis . Of a total of eight diVerent treatments with entomopathogenic nematodes in the glasshouse trial, three reduced the invasion of G. rostochiensis and one reduced the numbers of new cysts that were produced compared with controls. In the outdoor experiment, seven of the 12 treatments gave a reduction in invasion but none resulted in changes in the numbers of cysts found at plant senescence. In general, invasion of G. rostochiensis juveniles was reduced more eVectively by S. carpocapsae than by S. feltiae , and was greatest in the outdoor trial where larger inocula of entomopathogenic nematodes were used. Overall, the results indicated that use of S. feltiae and S. carpocapsae is unlikely to provide a viable control strategy for G. rostochiensis .     

Development of the one-on-one quality assessment assay for entomopathogenic nematodes.

The one-on-one bioassay was developed using Steinernema carpocapsae (All) nematodes against the wax moth larva, Galleria mellonella. The assay was used to develop and compare virulence profiles of both in vitro- and in vivo-produced nematodes and to provide a quality assessment 'standard' for in vitro-produced nematodes. The bioassay was subsequently used to develop virulence profiles for Steinernema carpocapsae (UK), S. feltiae (UK), S. feltiae (R1.5), S. feltiae (SN), S. glaseri (NJ-43), and S. riobrave (RGV). These profiles are unique for each species and isolate and are used as a standard of virulence in routine quality assessment of nematodes produced in liquid fermentation.     

昆虫病原线虫Steinernema longicaudum X-7增效药剂的筛选

测试了19种昆虫生长调节剂、植物源或病毒类药剂对昆虫病原线虫Steinernema longicaudumX-7的存活和侵染率的影响。筛选9种对线虫存活和侵染率均无影响的药剂,测定其与线虫混用对黄粉虫Tenebrio molitorL.9~11龄幼虫的交互作用。结果显示,各种参试药剂和线虫组合均表现为相加作用。为了进一步确定参试药剂与线虫混用对目标昆虫卵圆齿爪鳃金龟Holotrichia ovataChang 3龄幼虫的交互作用,本研究选择氟虫脲、除虫脲、灭幼脲、苦参碱、氟啶脲和虫酰肼在低浓度下与线虫混用,除虫脲与线虫混用对测定昆虫处理7 d后表现为增效作用,其它药剂与线虫则表现为相加作用;各种参试药剂与线虫组合处理14 d后均表现为相加作用。     

Laboratory and field evaluation of Korean entomopathogenic nematode isolates against the oriental beetle Exomala orientalis (Coleoptera: Scarabaeidae)

The oriental beetle Exomala (Anomala) orientalis (Waterhouse) is an important pest of turfgrass in Korean golf courses, and although a few chemical insecticides are registered for insect pest control, they are not very effective against scarab larvae. There is also a growing concern in Korea about the run-off of insecticides into sensitive habitats and the potential for groundwater contamination. A safe and environmentally sound alternative is needed to conventional insecticides. We therefore evaluated six Korean entomopathogenic nematode isolates: S. carpocapsae Pocheon, S. glaseri Dongrae, S. glaseri Mungyeong, S. longicaudum Gongju, S. longicaudum Nonsan, and Heterorhabditis sp. Gyeongsan for their potential as bioinsecticides for control of E. orientalis. In addition, we evaluated a reduced chemical insecticide approach that combined chlorpyrifos-methyl with nematodes. In laboratory tests Heterorhabditis sp. Gyeongsan was the most efficacious, causing 100% mortality of the second and 38% of the third instars. All other nematode isolates caused 50-80% mortality of the second and 15-30% of the third instars. E. orientalis pupae were highly susceptible to all the Korean entomopathogenic nematode isolates except S. carpocapsae. In artificially infested field plots, all Korean nematode isolates cause 50-70% mortality of the third instar. A combination of a one-half rate of Heterorhabditis sp. and a one-half rate chlorpyrifos-methyl was synergistic, causing 91% mortality compared with 69% for the full rate of Heterorhabditis sp. or 22% for the full rate of chlorpyrifos-methyl. In a second field trial, a natural infestation of preoverwintering third instar was treated. In this trial a one-half rate of S. longicaudum Nonsan plus a one-half rate of chlorpyrifos-methyl caused 96.8% mortality, much more than a full rate of S. longicaudum Nonsan (45.9% mortality) or a full rate of chlorpyrifos-methyl (28.7% mortality). The interactions of Heterorhabditis sp. and S. longicaudum Nonsan with chlorpyrifos-methyl in our field trials appear to be synergistic.     

Enhanced Trehalose Accumulation and Desiccation Survival of Entomopathogenic Nematodes Through Cold Preacclimation

Limited storage stability is a major obstacle to further expansion of the use of entomopathogenic nematodes for pest control. Progress has been made that Steinernema carpocapsae can now be stored under partial anhydrobiosis for up to 6 months at 25°C and 10 months at 5°C in a water-dispersible granular (WG) formulation. However, other species have been more difficult to store in the WG formulation due to migration of nematodes out of the granules and sensitivity of some species to desiccation directly at cold temperatures. As acclimation to cold induces trehalose accumulation (a major cryo- and desiccation protectant) in many invertebrates, it was hypothesized that cold preacclimation of entomopathogenic nematodes will enhance their survival in the WG formulation at cold temperatures. This hypothesis was tested using a temperate species Steinernema feltiae , a subtropical species S. carpocapsae , and a tropical species Steinernema riobrave possessing different thermal niche breadths and reproduction temperature optima. Cold acclimation of infective juveniles increased trehalose accumulation in all three species and the amount of trehalose accumulated was both temperature and species dependent. Trehalose content reached at its peak after 6 days at 5°C in S. feltiae (82.28 μg/mg dry weight), after 10 days at 10°C in S. carpocapsae (94.16 μg/mg dry weight) and after 6 days at 15°C in S. riobrave (47.58 μg/mg dry weight). Cold preacclimation at 5°C for 2 days enhanced desiccation survival of S. feltiae in 25% glycerol (osmotic desiccation) at both 5 and 25° and of S. carpocapsae and S. riobrave only at 5°C. Non-cold acclimated S. carpocapsae and S. riobrave were extremely sensitive to desiccation directly at 5°C in 25% glycerol, resulting in over 98% mortality within 6 days, but S. feltiae was more sensitive to desiccation at 25°C than at 5°C. Cold preacclimation increased survival of all the three species in the WG formulation at both 5 and 25°C. The survival of S. riobrave at 5°C in the WG formulation was positively correlated with the length of preacclimation period at 5°C (R 2 = 0.99) and with the amount of trehalose accumulated during cold preacclimation (R 2 = 0.81). These results support the hypothesis that cold preacclimation enhances desiccation survival of entomopathogenic nematodes at cold temperatures and the increased survival correlates well with the increased trehalose accumulation. Results also demonstrate that cold preacclimation can be used as a tool to enhance survival of nematodes in the formulations with reduced water activity.