Efficacy of Metarhizium anisopliae in controlling the two‐spotted spider mite Tetranychus urticae on common bean in screenhouse and field experiments

The efficacy of aqueous and emulsifiable formulations of the fungus Metarhizium anisopliae isolate ICIPE78 was evaluated on the population density of Tetranychus urticae infesting common bean plants under screenhouse and field conditions. Synthetic acaricide abamectin was included as a check. Bean plants were artificially infested with T. urticae and allowed to multiply. Three treatments were applied in the screenhouse and 1 treatment in field trials. Mite density was recorded 2 d before spraying and weekly postspraying. The number of pods per plant, number of seeds per pod, and the dry weight of seeds per plant were recorded only in the screenhouse trials. In both screenhouse and field trials, fungal formulations applied at the concentration of 10⁸ conidia/mL and the acaricide reduced the population density of mites as compared to the controls. There were significant differences in T. urticae population densities between the treatments at the various post‐spraying sampling dates. In the screenhouse, the mite densities were near zero from 3‐week postspraying in the treated leaves. At 4‐week postspraying, there were no more leaves in the untreated control (T1) and in the control water + Silwet‐L77 (T2). Fungal formulations were as effective as abamectin in reducing mite densities in both screenhouse and field experiments. There were significant differences in the production parameters during the 2 screenhouse trials, with fungal and abamectin treatments generally having the highest yield. Results of this study underline the potential of the M. anisopliae isolate ICIPE78 as an alternative to acaricides for T. urticae management.     

A fungal metabolite masks the host plant odor for the pine weevil (Hylobius abietis)

The pine weevil is one of the most important pest insects of conifer reforestation areas in Europe. Female pine weevils cover their eggs with chewed bark and feces (frass) resulting in avoidance behavior of feeding conspecifics towards egg laying sites. It has been suggested that microorganisms present in the frass may be responsible for producing deterrent compounds for the pine weevil. The fungi Ophiostoma canum, O. pluriannulatum, and yeast Debaryomyces hansenii were isolated from aseptically collected pine-weevil frass. The isolated fungi were cultured on weevil frass broth and their volatiles were collected by SPME and identified by GC–MS. D. hansenii produced methyl salicylate (MeS) as a major compound, whereas, in addition, O. canum and O. pluriannulatum produced 6-protoilludene. In a multi-choice lab bioassay, MeS strongly reduced pine weevil's attraction to the Pinus sylvestris volatiles. Thus, a fungal metabolite was found that strongly affects the pine weevil host-odor search.     

Development of methods for detection and Agrobacterium-mediated transformation of the sterile,endophytic fungus Muscodor albus

Symbiotic endophytes, unlike plant pathogens, do not usually induce visible host response. This may constraint the researcher's decision whether a plant has been successfully infected by the endophyte. In order to properly study the establishment, development and progress of an endophyte in the host plant and host-endophyte interactions, methods for the identification and localization of endophytic microorganisms are needed. Towards this aim, we focused at two levels: (A) We constructed M. albus-specific primers for polymerase chain reaction (PCR). In vitro, these primers specifically detected only M. albus strains and not isolates of related fungi (such as Daldinia sp. and a Xylariaceae sp.). (B) For direct visualization of the fungi, we inserted a reporter gene (gfp) into M. albus hyphae using Agrobacterium-mediated transformation. Since M. albus is a sterile fungus (i.e., without spores or fungal fruiting bodies), we used chopped fungal mycelium for the transformation procedure. We transformed three different isolates of M. albus using Agrobacterium-mediated transformation. Fifty-nine different transformants were collected with a transformation efficacy of 0.0004–0.0026%. Although PCR-based detection and direct visualization of the transformants in planta were unsuccessful, all tested transformants (with one exception) exhibited similar biological activity to their cognate wild type. This work provides a significant step forward in molecular research of the relationships between this endophytic genus and their hosts.     

哈尔滨市室外空气真菌组成和分布的初步研究

采用曝皿法(孟加拉红培养基)连续12个月对哈尔滨市实验林场、东北林业大学家属区和哈尔滨市二环路、兴路区间3个地点的空气真菌进行取样,初步确定了哈尔滨市室外空气真菌的种类及其动态变化。试验共得到9 908个真菌菌落,经鉴定为22属(或群)49种。试验结果表明:室外空气真菌的优势菌属依次为枝孢属、链格孢、曲霉属、青霉属、附球菌属、木霉属和无孢菌群,分别占总菌落数的21.54%、15.16%1、4.70%、12.20%、11.35%、7.86%和7.80%;室外空气真菌的种类在一天中基本没有变化,但菌落的数量有一定变化,总菌落数从早至晚也逐渐增加;不同季节室外真菌的种类和菌落数有明显变化,秋季最多,冬季最少;不同采样地点的空气真菌的种类变化不大,但菌落数量有差异,菌落数量依次为公路林场家属区。     

Distribution,occurrence and natural invertebrate hosts of indigenous entomopathogenic fungi of Central India

We report here that, during periodical surveys of insects inhabiting diverse habitats for the collection of entomopathogenic fungi; a large number of isolates were recovered belonging to seven species, from various regions of Madhya Pradesh and Chhattisgarh forest areas and agricultural fields. The most common entomopathogenic fungi such as Beauveria bassiana, Nomuraea rileyi, Paecilomyces farinosus and Paecilomyces fumosoroseus were found to infect various insect hosts species naturally viz. Hyblaea puera, Eutectona machaeralis, Diachrysia orichalcea, Spodoptera litura, and few new insect hosts of these fungal pathogens among Indian insect population were collected for the first time from Central India, such as beetles of Agrilus species, hairy caterpillars of Lymantria species. The isolation, identification, maintenance and pathogenicity assay of these isolates was performed prior to deposition in culture collection center.     

Genetic characterization of Mortierella alpina by sequencing the 18S‐28S ribosomal gene internal transcribed spacer region

Aims: The aim of this study was to establish a genetic background of Mortierella alpina, which is a cosmopolitan, soil‐inhabiting Zygomycete that also has important biotechnology potential. Methods and Results: A total of 44 18S‐28S ribosomal gene internal transcribed spacer (ITS1‐5·8S rDNA‐ITS2 DNA) regions of M. alpina from three diverse locations (Far East Asia, North America and West‐Central Europe) were sequenced and investigated. The sequences between M. alpina and the three closely related species (Mortierella macrocystis, Mortierella gamsii and Mortierella humilis) showed 74–84% sequence identity. When a phylogenetic tree was constructed with a neighbour‐joining algorithm, four clades of M. alpina isolates were clearly distinct with high bootstrap values. In addition, isolates from the West‐Central Europe were found to have the highest gene and nucleotide diversities. Conclusions: The ITS region was a suitable tool for distinguishing M. alpina from other closely related species. The region also provided information of the diversity of M. alpina. Significance and Impact of the Study: The study established a genetic background of M. alpina for identification and the diversity of M. alpina provided information for further isolation and screening of the fungus.     

Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii

Sclerotium rolfsii (strain CBS 350.80) was found to produce extraordinary high amounts of polygalacturonases (PGs). Two of these extracellular enzymes were purified by a recently introduced preparative electrophoretic device (isoelectric focusing mode of free flow electrophoresis). PG 1 (39.5 kDa, pI 6.5) and PG 2 (38 kDa, pI 5.4) exhibited quite similar properties, they were found to be both endo-acting enzymes. Both PGs cleaved penta- and trigalacturonic acid while tetragalacturonic acid was only cleaved when trigalacturonic acid was present. The latter substrate was hydrolysed much faster by PG 2. Both enzymes were active on pectins with different degrees of esterification, they were sensitive towards Ca-cations and not glycosylated. The kinetic properties were measured by viscosimetry with polygalacturonic acid as a substrate. NMR experiments on a model substrate revealed an inverting mechanism of carbohydrate hydrolysis for both enzymes.     

Probenazole-induced accumulation of salicylic acid confers resistance to Magnaporthe grisea in adult rice plants

Probenazole (PBZ) is the active ingredient of Oryzemate, an agrochemical which is used for the protection of rice plants from Magnaporthe grisea (blast fungus). While PBZ was reported to function upstream of salicylic acid (SA) in Arabidopsis, little is known about the mechanism of PBZ-induced resistance in rice. The role of SA in blast fungus resistance is also unclear. The recommended application period for Oryzemate is just before the Japanese rainy season, at which time rice plants in the field have reached the 8-leaf stage with adult traits. Thus, the involvement of SA in PBZ-induced resistance was studied in compatible and incompatible blast fungus-rice interactions at two developmentally different leaf morphology stages. Pre-treatment of inoculated fourth leaves of young wild-type rice plants at the 4-leaf stage with PBZ did not influence the development of whitish expanding lesions (ELs) in the susceptible interaction without the accumulation of SA and pathogenesis-related (PR) proteins. However, PBZ pre-treatment increased accumulation of SA and PR proteins in the eighth leaves of adult plants at the 8-leaf stage, resulting in the formation of hypersensitive reaction (HR) lesions (HRLs). Exogenous SA induced resistance in adult but not young plants. SA concentrations in blast fungus-inoculated young leaves were essentially the same in compatible and incompatible interactions, suggesting that PBZ-induced resistance in rice is age-dependently regulated via SA accumulation.     

栎金钱菌活性提取物抗衰老研究

用栎金钱菌活性提取物灌胃衰老模型小鼠,观察其对小鼠血清SOD活性、肝组织MDA和脂褐素的含量、脾指数、胸腺指数和肾指数的影响。结果表明,栎金钱菌活性提取物能使雄性小鼠血清SOD活性提高40.16%,肝MDA含量减少47.12%,肝脂褐素含量减少89.48%,脾指数增加35.13%,胸腺指数增加48.92%,肾指数增加11.26%,表明栎金钱菌活性提取物具有良好的抗衰老作用。栎金钱菌活性提取物800mg/kg·d的剂量具有最好的抗衰老效果,且具有性别的差异,对雄性小鼠的抗衰老作用略优于对雌性小鼠的。