土壤微生态环境对冬虫夏草寄主蝙蝠蛾幼虫病原菌粉棒束孢宿存及毒力的影响研究

测定了温度、含水量、土壤微生物三个因素以探讨蝙蝠蛾幼虫生长过程中微生态环境对粉棒束孢(Isaria farinose)宿存及毒力的影响。结果表明粉棒束孢30℃保存6个月的宿存量降低至10~3个/g以下。随着保存温度降低,宿存量的下降速度变缓,粉棒束孢在-18℃保存1年后,宿存量仍基本维持在10~6个/g以上。粉棒束孢在低湿环境下(含水量35%)保存1年的宿存量维持在10~4个/g,含水量45%～65%,粉棒束孢宿存量会先增加后降低,降低的速度基本随湿度的增加而增加,1年后宿存量降低至10~3个/g以下。土壤微生物对粉棒束孢的宿存具有明显抑制作用,野外高山草甸土壤中微生物显著抑制粉棒束孢萌发,不利于粉棒束孢的宿存,而室内饲养土壤由于幼虫排泄物、饲料腐殖质可能有利于粉棒束孢的宿存。上述研究结果为进一步制定粉棒束孢防治方法,同时也为粉棒束孢作为生防菌的有效保存提供了参考。     

Production and extraction of extracellular lipase from the entomopathogenic fungus Isaria fumosoroseus (Cordycipitaceae; Hypocreales)

Lipases are important cuticle degrading enzymes involved in the infection process of entomopathogens by hydrolysing the ester bonds of lipoproteins, fats and waxes present in the insect integument. Production of extracellular lipase by Isaria fumosoroseus (Cordycipitaceae; Hypocreales) isolate IF28.2 was investigated using different combinations of basal medium components. The effect of different vegetable oils added to a basal medium at different concentrations to improve enzyme production was evaluated. Maximum lipase activity (125.33±2.96 U/mL) as well as maximum biomass production (22.36±0.99 mg/mL) was observed for olive oil when used at a concentration of 2% (v/v) of the basal medium. In the presence of surfactants, the highest lipase activity occurred when SDS and Tween 80 were added at the time of fungal inoculation. SDS proved to be the best surfactant having 110.66±3.52 U/mL lipase activity. The effects of the divalent metal ions (iron and magnesium) on lipase activity were also studied. Iron inhibited, whereas magnesium slightly increased lipase activity. The optimum pH for lipase production was 5.7 while 32°C proved to be the best temperature for lipase production.     

Entomopathogenic fungi disturbed the larval growth and feeding performance of Ocinara varians (Lepidoptera: Bombycidae) larvae

Abstract Feeding experiments using three strains of entomopathogenic fungi, Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea were conducted with newly moulted 3rd–5th instar Ocinara varians Walker larvae in the laboratory. The mortality of larvae immersed individually in spore suspension (1 × 10⁷ spores/mL) of all the strains was ≥ 80% except 5th instar larvae treated with M. anisopliae which transformed into pupae, but did not result in adult emergence. The growth (total body mass), consumption, relative consumption rate and relative growth rate, were reduced at all three larval stages, while developmental time was extended in infected larvae with concurrent significant increase in approximate digestibility in infected larvae. Conversion of digested food (ECD) and ingested food (ECI) values declined in infected larvae as compared to the healthy larvae (control). The 5th instar larvae treated with M. anisopliae showed higher ECD and ECI values than control. Based on mortality and growth inhibition it can be suggested that all the studied fungal strains have a high potential for biocontrol and could be developed into biocontrol agents against O. varians.     

绿僵菌素对爪哇棒孢霉SP053菌株的影响及其混用对小菜蛾的联合毒力

为了寻找更有效的利用虫生真菌爪哇棒孢霉Isaria javanicus防治小菜蛾Plutella xylostella的新方法, 测定了绿僵菌素（destruxin）对SP053菌株生长发育与产孢量与萌发率的影响, 利用协同毒力指数法和毒力回归分析法评价了两者混用对小菜蛾2龄幼虫的联合毒力。结果表明: 绿僵菌素对SP053菌株菌丝生长、孢子萌发率和孢子产量均无显著影响（P>0.05）。而一定浓度的绿僵菌素与SP053菌株分生孢子混用具有增效作用, S50-CD100（指爪哇棒孢霉SP053菌株的分生孢子浓度为50×10⁵个/mL和绿僵菌素粗毒素的浓度为100 mg/L的组合; 依此类推）、S25-CD100、S25-CD50及S12.5-CD100等混配组合都有显著的增效效果, 尤其以S25-CD100组合增效效果最好, 其48 h和72 h的协同毒力指数（c.f.）分别达到52.31和31.07。毒力回归分析结果也显示, 添加绿僵菌素粗毒素25~100 mg/L的混剂的LC50值明显降低, 比如添加100 mg/L绿僵菌素粗毒素的混剂（SP053-CD100）, 其48和72 h的LC50分别为17.45和10.55 （×10⁵个孢子/mL）, 而SP053菌株分生孢子单剂的LC50相应值为>50和35.85（×10⁵个孢子/mL）。本研究证明绿僵菌素与爪哇棒孢霉SP053菌株混用有增效作用, 对改进小菜蛾的生物防治方法有一定指导意义。     

A Group I Intron in the 18S Ribosomal DNA from the Parasitic Fungus Isaria japonica

The nucleotide sequence of the 18S rDNA coding gene in the ascomycetes parasitic fungus Isaria japonica contains a group I intron with a length of 379 nucleotides. The identification of the DNA sequence as a group I intron is based on its sequence homology to other fungal group I introns. Its group I intron contained the highly conserved sequence elements P, Q, R, and S found in other group I introns. Surprisingly, the intron sequence of I. japonica is more similar to that of Ustilago maydis than to the one found in Sclerotinia sclerotiorum. This is in contrast to the sequence identity found on the neighboring rDNA. This is an interesting finding and suggests a horizontal transfer of group I intron sequences. Received: 19 September 1997 / Accepted: 10 September 1998     

Virulence of Hypocreales fungi to pecan aphids (Hemiptera: Aphididae) in the laboratory

There is need for efficacious biocontrol agents for aphids in commercial orchards. As a preliminary step to this end we determined the virulence of several Hypocreales fungi to pecan aphids. In the first experiment we tested the virulence of Isaria fumosorosea (ARSEF 3581) blastospores to three pecan aphids Monellia caryella, Melanocallis caryaefoliae, and Monelliopsis pecanis under laboratory conditions. Rates of 1 × 10⁷ or 1 × 10⁸ spores per ml were applied in 2 ml via a spray tower to 90 mm Petri dishes containing 10 aphids each. Mortality and mycosis were determined after 24, 48 and 72 h. Treatment effects were observed by 48 h post-application, and by 72 h the higher application rate caused >90% mortality and mycosis in M. caryella and M. caryaefoliae, whereas <70% was observed in M. pecanis.We conducted two subsequent experiments (Experiments 2 and 3), using the same methodology, to compare the virulence of several Hypocreales species and strains against the aphid of primary economic concern to most pecan growers, M. caryaefoliae. In Experiment 2, we compared blastospores and conidia of two I. fumosorosea strains (ARSEF 3581 and ATCC 20874 [= strain 97]). The blastospores of ARSEF 3581 and conidia of ATCC 20874 showed higher virulence than other treatments and thus were included in Experiment 3, which also compared the virulence of conidia of Beauveria bassiana (GHA strain) and Metarhizium anisopliae (F52 strain). Results in Experiment 3 indicated the highest virulence in I. fumosorosea 3581 blastospores and M. anisopliae (F52) followed by I. fumosorosea (20874) conidia. The detection of pathogenicity to pecan aphids establishes the potential for commercial usage and additional study. Results reported here will narrow treatments to test in future greenhouse and field trials.     

Light-induced fruit body formation of an entomogenenous fungusPaecilomyces tenuipes

Asexual fruit body formation of an entomogenous,Paecilomyces tenuipes, on an artificial culture medium was investigated. For the fruit body induction, light/dark condition was important. Illumination by white light in a particular critical period preceded by a dark period induced the fruit bodies. The critical period was identified to be around 6–14 d after inoculation at 20°C.     

Comparison of aerial conidia and blastospores from two entomopathogenic fungi against Diaphorina citri (Hemiptera: Liviidae) under laboratory and greenhouse conditions

This study compared the insecticidal activity of liquid culture-produced blastospores and solid substrate-produced aerial conidia of Beauveria bassiana GHA and Isaria fumosorosea ARSEF3581 strains against Diaphorina citri adults. Insects exposed to 10⁷ propagules/ml in a spray residue contact leaf bioassay died within 6 days at 25°C, with no significant differences between fungal treatments. At higher concentrations (10⁸ propagules/ml), I. fumosorosea conidia killed psyllids faster compared to its blastospore formulation, i.e. 4 versus 5 days, respectively. In greenhouse tests, the same treatments applied to infested citrus plants (2?×?10⁶ spores/ml) all significantly reduced the number of nymphs compared with the untreated controls over 3 weeks; however, only I. fumosorosea blastospores significantly reduced the number of F1 adult psyllids when compared with controls. Similar results were observed in the follow-up greenhouse test, where I. fumosorosea blastospores were the most effective treatment overall, reducing D. citri populations by about 60% after 21 days; by contrast, imidacloprid killed almost 100% of psyllids within a week in both tests. Fewer psyllids exhibited mycosis in the greenhouse (i.e. ≈20 versus?≥?87% in the laboratory). This is the first report comparing both conidial and blastospore formulations of B. bassiana and I. fumosorosea for the control of a psyllid pest. Field testing is required to determine how successful different spore formulations might be under various environmental conditions.     

温湿度对玫烟色棒束孢IF‐1106菌株孢子萌发及对烟粉虱致病力的影响(英文)

B型烟粉虱Bemisia tabaci Gennadius是一种重要的世界性农业害虫。玫烟色棒束孢Isaria fumosorosea是防治烟粉虱的一种重要的虫生真菌。对不同温度(20、23、26、29、32℃)、不同湿度(53%、65%、75%、85%、95%)下玫烟色棒束孢IF‐1106菌株分生孢子萌发及其对烟粉虱的致病力进行了测定。结果表明:温度对孢子的萌发有显著影响,26℃时孢子萌发率最高。当相对湿度低于75%时,孢子不萌发或萌发率较低;当相对湿度为85%–95%时,孢子萌发率显著升高。26℃时,烟粉虱2龄若虫的累计死亡率最高。随着相对湿度增大,病菌的致病力增强。当相对湿度为53%–95%时,烟粉虱2龄若虫累计死亡率从54.55%增加到88.89%。玫烟色棒束孢的致病力与孢子萌发率呈正相关,但温度比相对湿度对其影响更明显。结果表明,玫烟色棒束孢IF‐1106菌株侵染烟粉虱的最佳条件是温度26℃和相对湿度大于85%。     

Characterisation of Isaria fumosorosea isolates and their virulence toward the Diamondback Moth,Plutella xylostella

Some morphological and physiological characteristics of an Isaria fumosorosea isolate with diminished virulence, IFCF01-D, and its parent isolate, IFCF01, were evaluated and laboratory bioassays were performed to assess their virulence against Plutella xylostella. The relationship among these traits and virulence against P. xylostella is discussed. There were no significant differences in conidial viability, spore production and the time required for 50% germination (GT₅₀). Spore viability after incubation for 24 h at 25°C was greater than 98% for both isolates tested. Spore production on potato dextrose agar after 14 days incubation at 25°C was 4.68 × 10⁸ and 4.59 × 10⁸ conidia/mL for IFCF01 and IFCF01-D, respectively. When exposed to high temperatures (40, 45, 50 or 55°C) through a water bath for 10 min, conidial germination ranged from 0.83% to 84.0% for IFCF01 and 0% to 86% for IFCF01-D. Germination rate showed a negative relationship with the exposure temperature for both isolates. The per cent germination of isolate IFCF01 24 h after ultraviolet (UV) radiation (18 W, 240–260 nm) varied from 0% to 92% and 0% to 81% for IFCF01-D. Germination rate and the exposure time exhibited a negative correlation for both isolates tested. Conidial surface hydrophobicity of IFCF01 (60%) was significantly higher than that of isolate IFCF01-D (53%). Subsequently, using the cicada exuviae as the substrate for enzymatic analysis, Pr1 and chitinase activity demonstrated the contrasting virulence traits: higher specific activities for the more virulent IFCF01 and lower enzymatic levels for isolate IFCF01-D.