云南若干地区土壤放线菌区系及资源考察 X.滇东南地区

从云南省文山、红河等地采集不同植被、不同海拔高度的土壤样品,用不同方法分离高温、中温放线菌,按国内外通用的方法进行鉴定。滇东南地区的土壤放线菌种类丰富,分离到了13个菌属,稀有放线菌的数量所占比例较高。原始森林中的放线菌种属最丰富。本文对该地区放线菌的生态分布进行了探讨。     

Differentiation and protease production in Micromonospora echinospora (ATCC 15837)

Micromonospora echinospora differentiates in both submerged and surface cultures producing abundant dark spores after a period of vegetative mycelial growth. In submerged batch cultures, under either carbon or nitrogen limiting conditions, protease activity was found to coincide with sporulation indicating a relationship between proteolytic activity and differentiation in this organism. Further evidence for this link was provided from surface grown cultures wherein sporulation was inhibited by the serine protease inhibitors TLCK and TPCK. The association between proteolysis and differentiation apparent in this organism correlates with evidence of a similar phenomenon observed in the streptomycetes, suggesting that this may be a common response associated with differentiation in filamentous actinomycetes.     

云南若干地区土壤放线菌区系及资源考察——Ⅸ.滇西和滇东北地区

云南省西部属横断山脉的南端,该地区内温泉分布十分广泛。云南省的东北部则属中山山原区,是乌蒙山系向北伸延的末端。这两个地区的地貌、气候、植被区系均有很大差异。从上述两个样区的原始森林、次生林、荒地、旱地、蔬菜地、水田采集200份土样,用多种培养基分离和鉴定其中的放线菌,并从中筛选出有用的菌株,研究了上述两地区的土壤放线菌的区系组成及异同。     

The influence of fertilizers on root rot of field peas caused by Fusarium oxysporum,Pythium vexans and Rhizoctonia solani inoculated singly or in combination

A Frankia strain ISU 0224887 was isolated from spore negative root nodules of Gymnostoma sumatranum and was grown in pure culture. It was infective and effective for Gymnostoma species but failed to nodulate Allocasuarina and Casuarina seedlings. Light and scanning electron microscopy of it in nitrogen free medium revealed a filamentous mat of septate and branched hyphae bearing sporangia and vesicles capable of fixing nitrogen. The strain also produced an orange pigment after 2 weeks culture. The strain utilized only TWEEN 80 and propionate as sole carbon sources. The different antibiotics used showed varying effects on its growth.     

Endophyte sporulation in root nodules of actinorhizal plants

All strains of isolated Frankia possess the genetic capacity to form sporangia since, when grown in vitro, they usually sporulate freely, depending on the physical and chemical environment in which they are cultured. Endophytic sporulation involving Frankia differentiation of sporangia within root nodules has been described in only 16 host species in 9 genera within six families of actinorhizal plants. From studies published to date, endophytic sporulation cannot be correlated with specific environmental conditions surrounding the host plants. Based on the literature and on previously unpublished observations from field and greenhouse studies, an account is given of the occurrence of sporulation in actinorhizal plants with emphasis on Alnus, Casuarina, Comptonia, Elaeagnus and Myrica . The possible role of the host plant in controlling Frankia sporulation as contrasted to the control exerted by the genetic constitution of the microbial symbiont is explored.     

Nitrogen-fixing root nodules inDatisca Cannabina L.

Summary The occurrence of root nodules inDatisca cannabina is reported. The nodules are typically of the Alnus type, forming dichotomously-branched coralloid clusters. The enlarged cortical cells contain vesicle clusters of the endophyte. The nodules reduced acetylene to ethylene at a rate of 5.5 moles ethylene per g fresh wt of nodules per hour.     

An actinomycete producing L-3,4-dihydroxyphenylalanine from L-tyrosine

An actinomycete was isolated during a soil screening programme to obtain L-3,4-dihydroxyphenylalanine producers. A mutant of this organism was isolated by chemical mutagenesis and it accumulated 1 g/litre L-dihydroxyphenylalanine when grown on L-tyrosine. Resting cells converted 30% of tyrosine in the reaction mixture. The use of resting cells for dihydroxyphenylalanine production is advantageous as it eliminates interfering substances which accumulate during fermentation.     

β-Lactamases from Actinopolyspora halophila,an extremely halophilic actinomycete

Actinopolyspora halophila, an extremely halophilic actinomycete, produced both cellular and exocellular -lactamases when grown in liquid media containing 20% (w/v) NaCl. Maximal exocellular -lactamase production occurred at 48 h growth and exceeded cellular enzyme levels 7-fold. Maximal cellular -lactamase was observed at 72 h as cultures achieved full growth.Both -lactamases were purified to molecular homogeneity by a sequence involving gel filtration on Bio-Gel P-100, DEAE Bio-Gel A chromatography, preparative isoelectric focusing, and gel filtration on Sephadex G-75. Cellular -lactamase was purified 99-fold with 12% recovery and had a molecular weight of 42,200, and an isoelectric point of 4.15. Exocellular -lactamase was 12-fold purified with 1.2% recovery of initial activity and had a molecular weight of 38,000 and an isoelectric point of 3.85. Its specific activity was 7-fold greater than that of the cellular enzyme.A variety of penicillin and cephalosporin substrates were degraded by both enzymes. While the cellular -lactamase degraded phenoxymethylpenicillin, methicillin, and cephaloglycin most efficiently, the exocellular enzyme was most active against methicillin, carbenicillin, ampicillin, and cephalosporin C. Both enzymes were stimulated and protected from thermal deactivation by NaCl but not KCl or MgCl₂-Neither enzyme was inhibited by iodine.Abbreviations IEF Isoelectric focusing - SDS-PAGE sodium dodecylsulphate-polyacrylamide electrophoresis - PEG polyethyleneglycol - DTT dithiothreitol - DEAE diethylaminoethyl Issued as NRCC 25164     

辐射污染区土壤中放线菌的分离及多样性

从辐射污染区采集42份土样, 分别采用6种分离培养基进行放线菌的分离, 共获得152株放线菌。经形态、核糖体DNA扩增片段限制性内切酶(Amplifed ribosomal DNA restriction analysis, ARDRA)分析比较, 选取其中的60株进行16S rRNA基因测序。通过序列比对、聚类分析, 60株菌分布在放线菌纲中的12个属, 链霉菌属占大多数, 有大量稀有放线菌, 这表明辐射污染区具有较丰富的放线菌属种多样性。     

Isolation and characterization of actinobacteria ectosymbionts from Acromyrmex subterraneus brunneus (Hymenoptera, Formicidae)

The ectosymbiont actinobacterium Pseudonocardia was isolated from the integument of Acromyrmex leaf-cutter ants and seems to play a crucial role in maintaining asepsis of the nest. Currently, there has been an intensive search for Pseudonocardia associated with several attine species, but few studies have indicated that other actinobacteria may be associated with these ants as well. We therefore characterized the culturable actinobacteria community associated with the integument of the fungus-growing ant Acromyrmex subterraneus brunneus Forel, 1893 (Hymenoptera: Formicidae). Ectosymbionts were isolated using four different media and characterized by morphological and molecular (16S rDNA) methods. A total of 20 strains were isolated, of which 17 were characterized as Streptomyces spp., and one isolate each as Pseudonocardia, Kitassatospora and Propionicimonas. Unlike other Acromyrmex species, A. subterraneus brunneus is associated with a diversity of actinobacteria. Even though Pseudonocardia is present on this leaf-cutting ant’s integument, the number and diversity of Streptomyces spp. found differs from those of previous studies with other attine ants and suggest that different culturing approaches are needed to characterize the true diversity of microbes colonizing the integument of attine ants. Moreover, understanding the diversity of the culturable actinobacteria associated with A. subterraneus brunneus should increase our knowledge of the evolutionary relationship of this intricate symbiotic association.