Bioassay method for assessing the virulence of Beauveria bassiana against tarnished plant bug, Lygus lineolaris (Hem., Miridae)

A standard bioassay method for assessing the pathogenicity of Beauveria bassiana (Balsamo) Vuillemin (GHA strain) against second instar tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hem., Miridae) was developed. Several types of inoculation methods, assay containers and incubation times were tested. Our goal was to minimize control mortality and maximize treatment mortality. Five inoculation methods (immersing broccoli florets or bean pods, spraying broccoli florets or bean pods, and immersing insects) and four types of plastic containers (114‐, 171‐, 228‐ and 455‐ml) were tested. Immersing insects directly in a fungal suspension was the most effective inoculation method, which resulted in a treatment mortality of 70–81.3% at a concentration of 1 × 10⁷ conidia/ml. The 114‐ml plastic container was the most suitable assay container when 10 tarnished plant bug nymphs were treated together, resulting in a control mortality of only 6% 12 days after treatment. Within the first 6 days after treatment, 71.1% of the insects were killed, compared with a total mortality of 81.3% after 12 days. Nymphs infected with the test fungus changed colour from green to black. Mycelial outgrowth and sporulation on the cadavers demonstrated that most nymphs died of fungal infection. A total of 61.1 and 80.5% of the cadavers showed signs of mycelial outgrowth 9 days after death among those that were surface sterilized and those that were not, respectively.     

绿盲蝽成虫的田间活动规律

本文通过粘虫板诱捕试验,研究了绿盲蝽Apolygus lucorum(Meyer-Dür)成虫在田间的活动规律。结果发现,绿盲蝽成虫主要在棉花顶部附近飞行,在高于棉花顶部20 cm处诱捕到的绿盲蝽显著多于50 cm和80 cm处(P<0.05)。在田间飞行时,绿盲蝽成虫没有明确的偏好方向,在东西南北各方位上的成虫诱捕量没有显著差异(P>0.05)。在昼夜节律上,绿盲蝽成虫集中在傍晚至凌晨时分活动,其中16:00至翌日4:00之间雌雄成虫的诱捕量分别占全天的100%和92.6%。     

Detection of European Peristenus digoneutis loan in mirid populations in southern Ontario

Lygus spp. Hahn (Hemiptera: Miridae) are serious pests of a wide variety of economically important crops in North America. A European parasitoid, Peristenus digoneutis Loan (Hymenoptera: Braconidae), released in the northeastern USA for the biological control of Lygus, has successfully established in nine states and in eastern Canada, including southeastern Ontario, southern Quebec and Nova Scotia. To determine the extent to which P. digoneutis has dispersed and established in mirid populations in Ontario, a single-step multiplex PCR assay, designed to differentiate European species P. digoneutis and P. relictus from native North American Peristenus species, was used to identify parasitoid larvae dissected from field-collected Miridae. A total of 222 parasitoid larvae were analysed with the multiplex assay. Most (172) were identified as members of the P. pallipes Curtis complex; however, three from the Niagara region were identified as P. digoneutis. Specimens that did not amplify using the Peristenus multiplex PCR assay were screened with a PCR primer set designed to detect Leiophron (Hymenoptera: Braconidae) species, and most were confirmed to be Leiophron larvae. This study confirms the presence of P. digoneutis species in southern Ontario, and is another example of the utility of molecular methods for the detection of newly introduced or dispersed parasitoids. The presence of exotic P. digoneutis in southern Ontario may expedite future releases to augment already-established populations. Following such releases, the Peristenus multiplex PCR assay will be a useful component of post-release studies to evaluate the success of the biological control programme for Lygus plant bugs in Canada.     

绿盲蝽和中黑盲蝽对不同抗性和虫害处理棉花的选择趋性

研究绿盲蝽Apolygus lucorum和中黑盲蝽Adelphocoris suturalis对不同抗性和虫害处理棉花的选择趋性,为棉盲蝽的综合治理提供理论依据。本研究以抗性棉花品种BR-S-10（抗性品种）和感性棉花品种科林08-15（感性品种）,以及绿盲蝽和中黑盲蝽分别危害处理后的该两种棉花植株为试验材料,以纯净空气为空白对照,共成对设置15个气味源组合,采用“Y”型昆虫嗅觉仪室内研究了绿盲蝽和中黑盲蝽对不同抗性和虫害处理棉花的选择趋性。结果表明,绿盲蝽显著选择感性品种、中黑盲蝽危害感性品种和绿盲蝽危害感性品种。中黑盲蝽显著选择绿盲蝽危害感性品种、中黑盲蝽危害感性品种和感性品种。总的来说,两种棉盲蝽趋向于选择敏感棉花品种,抗性棉花品种对供试昆虫有显著趋避性;绿盲蝽显著趋向选择中黑盲蝽危害感性品种和绿盲蝽危害感性品种,中黑盲蝽显著趋向选择绿盲蝽危害感性品种和中黑盲蝽危害感性品种。     

棉花叶片性状（厚度和油点密度）与其对绿盲蝽抗性的关系

为了明确棉花叶片性状（厚度和油点密度）与其对绿盲蝽Apolygus lucorum （Meyer-Dür）抗性的关系,以期为棉花抗盲蝽品种(系)选育提供理论依据,本研究选用对绿盲蝽有不同抗性的20个棉花品种(系)为材料,分别测定其叶片厚度、表皮层厚度和油点密度。结果表明：晋棉26、冀丰989、常D、亚洲棉对绿盲蝽表现为高抗;棉花叶片厚度、表皮层厚度和油点密度在不同生长期均有差异;棉花对绿盲蝽的综合抗性指数与其铃期叶片表皮层厚度和油点密度有显著相关性（y=20.673+0.415x,R²=0.305,P=0.012;y=1.049+0.049x,R²=0.207,P=0.044）。结果显示,在一定的生长期,叶片表皮层厚和多油点棉花品种可以减少绿盲蝽的危害。     

Coating Beauveria bassiana with lignin for protection from solar radiation and effects on pathogenicity to Lygus lineolaris (Heteroptera: Miridae)

The entomopathogenic fungus, Beauveria bassiana, is highly susceptible to the damaging effects of solar radiation. This study demonstrates protection from simulated solar radiation by coating B. bassiana (GHA) spores with lignin and effects on pathogenicity to Lygus lineolaris (Palisot de Beauvois) adults in direct spray and contact uptake bioassays. Spores were coated with either lignin or cross-linked lignin by spray drying to produce spore coatings of high and low water solubility, respectively. Non-coated spores and the two spore coating treatments were suspended in either water (0.04% Silwet L77) or oil (Orchex 692) to produce six formulations. Rates of loss in spore viability under simulated solar radiation were approximately ten times lower for the three formulations in which spores remained coated in suspension (cross-linked lignin in water, cross-linked lignin in oil, and lignin in oil). However, these three formulations were the least pathogenic. Estimates of LC₅₀ and LC₇₅ values for the most pathogenic formulation (non-coated in oil) were 5 and 12 times lower, respectively, than the least pathogenic formulation (cross-linked lignin in oil), but these differences were not significant. The three lignin-coated formulations that provided the greatest UV protection were also the least pathogenic based on LT values, which were significant. Overall, L. lineolaris mortality was approximately 80 times less when exposed to treated broccoli rather than sprayed directly, based on LC₅₀ values. If the contribution of spore uptake from plant surfaces to L. lineolaris mortality is similarly low under field conditions, then improving persistence may be less important for improving efficacy. However, under control conditions where solar radiation significantly impacts mycoinsecticide efficacy, the improved persistence of lignin coating formulations may outweigh negative effects on pathogenicity.     

Factors affecting progeny production of Anaphes iole

Anaphes iole Girault(Hymenoptera: Mymaridae) is a native, solitaryegg parasitoid of Lygus spp.(Heteroptera: Miridae) in North America. Current research is considering factors thatmay optimize the in-vivo rearing of A.iole using Lygus hesperus Knight(Heteroptera: Miridae) as host. The effects ofhost density, day of oviposition and foodpresence, parasitoid age and mate presence onproduction of A. iole progeny weredetermined in this study. After exposingindividual parasitoids to host patches for 24h, the percentage of hosts containing a latestage A. iole pupa wassignificantly greater at a moderate hostdensity (41–70 eggs per patch) than at a lowdensity (10–40 eggs). But, no differenceswere detected between moderate and high density(71–100 eggs) or between high and low densitytreatments. More adult progeny were generatedby females (of variable age; 0–2 d old) onthe first day of oviposition rather than thesecond day, regardless of food presence. Progeny sex ratio was decidedly male-biased onthe second day. Female age (0 d vs 1 d old)had a marginal effect on production in 24 h; 0d old females tended to generate more adultprogeny than 1 d old females. Overall, thisresearch suggests that exposing newly-emerged,unfed, mated A. iole females to amoderate to high host density for 1 to 2 dcould lead to time-efficient production ofadult progeny in an in-vivo rearing system.     

两种室内生物测定方法评价杀虫剂对绿盲蝽的相对毒力

随着Bt棉在我国的广泛种植, 绿盲蝽Lygocoris lucorum Meyer-Dür的危害日益加重。化学防治作为棉田盲蝽综合治理的重要策略, 仍是其主要的防治手段。本研究主要用闪烁管药膜法和点滴法两种测定方法评价了传统杀虫剂、新型杀虫剂等30种杀虫剂对绿盲蝽成虫的急性触杀毒力, 其中多种杀虫剂是首次使用两种生测方法同时进行毒力测定。结果表明：有机氯类、有机磷类、氨基甲酸酯类和拟除虫菊酯类等传统杀虫剂对绿盲蝽成虫具有较高的触杀活性。8种对绿盲蝽成虫具有较高毒力的药剂, 其中5种为传统杀虫剂, 分别为马拉硫磷、毒死蜱、联苯菊酯、灭多威和硫丹。新型吡咯类杀虫剂对绿盲蝽表现出较高毒力, 以氟虫腈的毒力最高, 两种生测方法测定的LC₅₀分别为0.13 μg·mL^-1 和 0.15 μg·mL^-1。而昆虫生长调节剂、抗生素类及植物源杀虫剂对绿盲蝽成虫没有明显的触杀活性。     

绿盲蝽及其天敌在不同生态条件下的发生动态研究

2005年～2007年,在山东省德州市农科院良种场棉田内系统研究了不同生境对抗虫棉绿盲蝽及其天敌发生动态的影响。试验设5个试验10种处理：即远离果园棉、紧靠果园棉,亩株数3600株均行、亩株数3000株并大小行种植棉,合理化控棉、不化控棉,纯作棉田和插种葵花棉田,双基因抗虫棉和单基因抗虫棉。在棉花的整个生育期系统调查不同生境棉田主要害虫及其天敌的种群数量。结果表明,不同生境抗虫棉田内绿盲蝽天敌的种类基本相同,主要为龟纹瓢虫、七星瓢虫、中华草蛉、大草蛉、华姬蝽、小花蝽、T-纹豹蛛、三突花蛛、草间小黑蛛等。不同生境抗虫棉田内绿盲蝽及其天敌种群数量差异明显。远离果园棉、低密度亩株数3000株并大小行种植棉、合理化控棉能减轻绿盲蝽的发生,但不利于绿盲蝽天敌的发生;种植双基因抗虫棉能减轻绿盲蝽的发生且对绿盲蝽天敌影响不大;插种葵花的棉田即能增加绿盲蝽天敌的数量,又能减轻绿盲蝽的发生。研究还表明绿盲蝽及其天敌在远离果园棉、紧靠果园棉下发生趋势基本一致,仅是发生量存在明显差异,远离果园棉田的绿盲蝽及其天敌百株发生量显著低于紧靠果园棉田（P〈0.05）;合理化控棉田、亩株数3000株大小行棉田内的绿盲蝽及其天敌发生数量显著低于不化控棉田、亩株数3600株均行棉田内的发生数量（P〈0.05）;双基因抗虫棉田内的绿盲蝽百株发生量显著低于单基因抗虫棉田发生数量（P〈0.05）;插种葵花棉田内的绿盲蝽百株发生量极显著低于纯作棉田（P〈0.01）、插种葵花棉田内的绿盲蝽天敌百株发生量显著高于纯作棉田（P〈0.05）,葵花对绿盲蝽及其天敌的诱集效果非常显著,在转基因棉田插种葵花是保益灭害控制绿盲蝽的有效方法之一。     

Characterization of Beauveria bassiana and Metarhizium anisopliae isolates for management of tarnished plant bug,Lygus lineolaris (Hemiptera: Miridae)

Selected morphological and physiological characteristics of four Beauveria bassiana (Balsamo) Vuillemin isolates and one Metarhizium anisopliae (Metschnikoff) Sorokin isolate, which are highly pathogenic to Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), were determined. There were significant differences in conidial size, viability, spore production, speed of germination, relative hyphal growth, and temperature sensitivity. Spore viability after incubation for 24h at 20 degrees C ranged from 91.4 to 98.6% for the five isolates tested. Spore production on quarter-strength Sabouraud dextrose agar plus 0.25% (w/v) yeast extract after 10 days incubation at 20 degrees C ranged from 1.6x10(6) to 15.5x10(6)conidia/cm(2). One B. bassiana isolate (ARSEF 1394) produced significantly more conidia than the others. Spore germination was temperature-dependant for both B. bassiana and M. anisopliae. The time required for 50% germination (TG(50)) ranged from 25.0 to 30.9, 14.0 to 16.6, and 14.8 to 18.0h at 15, 22, and 28 degrees C, respectively. Only the M. anisopliae isolate (ARSEF 3540) had significant spore germination at 35 degrees C with a TG(50) of 11.8h. A destructive sampling method was used to measure the relative hyphal growth rate among isolates. Exposure to high temperature (40-50 degrees C) for 10min had a negative effect on conidial viability. The importance of these characteristics in selecting fungal isolates for management of L. lineolaris is discussed.