Infection of Sentinel Cotton Aphids (Homoptera: Aphididae) by Aerial Conidia ofNeozygites fresenii(Entomophthorales: Neozygitaceae)

Uninfected adultAphis gossypii(Homoptera: Aphididae) apterae (sentinel aphids) on cotton leaves were exposed for 8 h to the air over a commercial cotton field in Louisiana during the night of 1–2 July 1995. At 0015 h there were 90,437 primary conidia/m³air of the fungusNeozygites fresenii(Entomophthorales: Neozygitaceae) at the midfield position as determined from Rotorod samples. Forty-eight percent (n = 106) of the sentinel aphids exposed for 8 h at midfield were infected by aerial conidia ofN. fresenii.Exposure of sentinel aphids outside the cotton field, at 10 and 100 m downwind and 10 m upwind, resulted in 34.8% (n = 131), 24.0% (n = 129), and 17.4% (n = 146) infected aphids, respectively. These data demonstrate that wind-dispersed aerial conidia ofN. freseniiare infective and rapidly and efficiently disperse the pathogen throughout aphid populations within and between fields.     

红火蚁病原真菌AUGM47早期发育超微结构研究

在前期筛选已获得对红火蚁Solenopsis invicta Buren高效致病真菌罗伯茨绿僵菌Metarhizium robertsii AUGM47的基础上,为进一步明确病原真菌对寄主昆虫的侵染机制。本试验在室内条件下,以红火蚁工蚁为侵染对象,利用荧光显微镜和透射电镜观察了罗伯茨绿僵菌AUGM47侵染单元分生孢子在体表附着萌发、穿透和体内增殖的早期发育过程。结果表明,菌株AUGM47分生孢子在红火蚁体表可萌发并形成附着胞侵入,接种后12 h观察到萌发,在36 h内普遍出现穿透结构穿透体壁。接种后48 h为菌体在血腔内的增殖阶段。菌丝体在穿透表皮和体腔内增殖过程中伴随着机械压力和酶的活动。接种后96 h,观察到自噬现象,菌体通过自噬降解并回收细胞器,为从体内穿出的晚期发育过程提供物质基础。本研究对罗伯茨绿僵菌AUGM47分生孢子在红火蚁体外至体内的发育进程研究证实了菌株的高致病性,为红火蚁生防真菌菌种改良和后续开发利用奠定理论基础。     

Effects of chemical and botanical insecticides used for locust control on Metarhizium anisopliae var. acridum conidia after short- to medium-term storage at 30°C

The short- to medium-term viability and growth of Metarhizium anisopliae var. acridum conidia were investigated when combined with six insecticides, at three different concentrations. All of the insecticides used in this study were suitable for immediate spraying with M. anisopliae var. acridum conidia except for fenitrothion. Fipronil, teflubenzuron, and fenitrothion formulations significantly reduced conidial viability over time. The 10% teflubenzuron treatment caused loss of viability relatively quickly with 9.9% germination after 28 days. Mycelial growth was affected by all the treatments except fenitrothion.     

Effect of temperature on growth, germination, germ-tube extension and survival of Paecilomyces lilacinus strain 251

Paecilomyces lilacinus and in particular the commercial strain 251 has been intensively tested for biological control of plant parasitic nematodes. Since this species has been mentioned in a number of reports concerning infection of humans, the human health risk for Paecilomyces lilacinus strain 251 was investigated. The effects of time, temperature and growth medium on radial colony growth and germination were determined. Additionally, exposure to 36°C and its effect on germ-tube extension and on survival of conidia was evaluated. Radial growth was significantly affected by temperature, growth medium and their interaction. Optimum temperatures were between 24 and 30°C, but no growth was found at 36°C. Germination rate was significantly influenced by time, medium, temperature and their interactions. The optimum temperature range for germination was between 28 and 30°C. Formulated conidia were capable of germinating at 36°C. However, studies on germ-tube extension conducted at 36°C showed a delay in development for 28-49 h and no further germ-tube extension was found after exposure for 80-95 h. Slopes of survival curves were significantly influenced by the type of conidia tested. In general, conidia did not survive exposure to 36°C for 168 h. These experiments indicate the temperature conditions where the strain is likely to be active and provide supporting data for full environmental and health risk assessments of biocontrol fungi.     

First evidence of ethylene production by Fusarium mangiferae associated with mango malformation

Malformation is arguably the most crucial disease of mango (Mangifera indica L.) at present. It is receiving great attention not only because of its widespread and destructive nature but also because of its etiology and control is not absolutely understood. Recently, Fusarium mangiferae is found to be associated with mango malformation disease. There are indications that stress ethylene production could be involved in the disease. Here we have shown the first direct evidence of production of ethylene in pure culture of F. mangiferae obtained from mango. The study also revealed that all the isolates dissected from mango acquire morphological features of F. mangiferae showing most similarity to the features of species with accepted standard features. The isolates of F. mangiferae from mango were observed to produce ethylene in significant amounts, ranging from 9.28–13.66 n mol/g dry wt/day. The findings presented here suggest that F. mangiferae could contribute to the malformation of mango by producing ethylene and probably stimulating stress ethylene production in malformed tissue of mango. Ethylene might be produced through 2-oxoglutarate-dependent oxygenase-type ethylene-forming-enzyme (EFE) pathway in Fusarium sp, which needs to be investigated.     

Inhibition of Farnesoic Acid Methyl transferase by Sinefungin

Sinefungin inhibited the S-adenosylmethionine-dependent farnesoic acid methyltransferase in a cell-free system containing a homogenate of corpora allata from female locusts, Locusta migratoria. The enzyme catalyzed the penultimate step of juvenile hormone biosynthesis in the insects. Culturing corpora allata in the presence of sinefungin greatly suppressed juvenile hormone production. The following in vivo effects were visible after injection of the inhibitor: increase in mortality and reduction of total haemolymph protein titer and ovary fresh weight, as well as length of terminal oocytes. Attempts to reverse these effects by topical application of the juvenile hormone analog ZR-515 (methoprene) were only partly successful. Therefore, the in vivo effects may be due to a general inhibition of methyltransferase enzymes in the insect. Sinefungin appeared to be of potential interest as the first representative of a new class of insect growth regulators.     

Effects of media composition on submerged culture spores of the entomopathogenic fungus, Metarhizium anisopliae var. acridum, Part 1: Comparison of cell wall characteristics and drying stability among three spore types

Metarhizium anisopliae var. acridum (IMI 330189) can produce at least three spore types in vitro; blastospores, submerged conidia, and aerial conidia, as defined by culturing conditions, sporogenesis, and spore morphology. This study compares morphological characteristics (dimensions and cell wall structure), chemical properties of cell wall surfaces (charge, hydrophobicity, and lectin binding), and performance (germination rate and drying stability) among these three spore types. Submerged conidia and aerial conidia both possessed thick, double-layered cell walls, with hydrophobic regions on their surfaces. However, in contrast to aerial conidia, submerged conidia have: (1) a greater affinity for the lectin concanavalin-A; (2) more anionic net surface charge; and (3) a less distinct outer rodlet layer. Blastospores were longer and more variable in length than both submerged conidia and aerial conidia, and had thinner single-layered cell walls that lacked an outer rodlet layer. Also, blastospores had a greater affinity than either conidia type for the lectin, wheat germ agglutinin. Blastaspores lacked hydrophobic regions on their surface, and had a lower anionic net surface charge than submerged conidia. In culture, blastospores germinated the fastest followed by submerged conidia, and then aerial conidia. Survival of submerged conidia and aerial conidia were similar after drying on silica gel, and was greater than that for blastospores. We provide corroborating information for differentiating spore types previously based on method of production, sporogenesis, and appearance of spores. These physical characteristics may have practical application for predicting spore-performance characteristics relevant to production and efficacy of mycoinsecticides.     

害虫防治用玫烟色拟青霉分生孢子粉的干燥工艺优化*

用液—固两相法生产的玫烟色拟青霉Paecilomyces fumosoroseus Pfr116菌株的分生孢子粉,在高真空冷冻干燥、高真空室温抽干、35℃下烘干和低真空低热干燥条件下进行不同程序的干燥处理,以筛选适合该菌孢子粉生产的干燥工艺条件。结果表明,低真空(0.1 MPa)低热(30℃)抽干20~24 h的干燥方法最适合用于该菌孢子粉的干燥,既能保证含水量在9.0%以下,又能保证87%以上的活孢率和1130亿~1310亿/g的含孢量,而且操作简便,成本较低,可作为高纯度孢子粉生产的首选干燥工艺。高真空(15.86 Pa)条件下无论冻干还是室温抽干,虽然孢子粉的含水量(2.2%~8.7%)和含孢量(1270亿~1360亿/g)指标符合生产要求,但活孢率仅62%,说明该菌孢子不适合在高真空条件下干燥。在35℃下烘干24 h所获孢子粉含水量、24 h萌发率和含孢量分别为9.6%、82.8%和1200亿/g,该方法也可在生产中应用,但其活孢率显著低于(P<0.05)低真空低热抽干24 h的孢子粉。     

Release of the extracellular matrix from conidia ofBlumeria graminis in relation to germination

The release of extracellular matrix (ECM) and the emergence of germ tubes from conidia ofBlumeria graminis were studied by light microscopy and micromanipulation. More prompt and frequent ECM release was confirmed on an artificial hydrophobic substratum than on an artificial hydrophilic substratum. Conidia initially incubated on the hydrophilic substratum were transferred by micromanipulation to either the hydrophobic or the hydrophilic substrata. Immediately after transfer onto the hydrophobic substratum, 75% of conidia released ECM, whereas only 16% did so upon transfer to the hydrophilic substratum. Conidia transferred onto the hydrophobic substratum produced a primary germ tube (PGT) more promptly and frequently than those transferred to the hydrophilic substratum. Thus, conidia recognize and respond to substratum hydrophobicity perhaps immediately after contact. When inoculated onto either isolated barley cuticle or the hydrophobic artificial substratum, 2/3 of the conidia produced a PGT from their polar regions. By contrast, on the hydrophilic substratum 2/3 of conidia did so from the side region. These results show that substratum hydrophobicity affects the location of PGT emergence from conidia. Furthermore, the study indicates that very rapid recognition of surface hydrophobicity by conidia promotes ECM release and this in turn may influence the location of PGT emergence.     

Pathogenicity, formulation and storage of insect pathogenic hyphomycetous fungi tested against Diabrotica speciosa

Studies were conducted tosearch for fungal strains with potentialpathogenicity against Diabrotica speciosa(Germar) (Coleoptera: Chrysomelidae).Among sixteen fungal isolates screenedthe most virulent was a Beauveria bassiana(Balsamo) Vuillemin isolate (FHD13) thatcaused 70% mortality of D. speciosathird instar larvae. The LC₅₀ value ofB. bassiana isolate FHD13 was3.48 × 10¹⁰ conidia/ml.Different temperatures (4, 17 and 26 °C)and vegetable oils (corn, sunflower and canola)used for storage did not significantly affectviability of conidia. A pathogenicity trialagainst D. speciosa larvae performed withthe corn oil formulation (1 × 10⁸ conidia/mlof oil) caused 65% of mortality.