Studies on the microbial populations of the rhizosphere of big sagebrush (<Emphasis Type="Italic">Artemisia tridentata</Emphasis>)

Prolonged use of broad-spectrum antibiotics has led to the emergence of drug-resistant pathogens, both in medicine and in agriculture. New threats such as biological warfare have increased the need for novel and efficacious antimicrobial agents. Natural habitats not previously examined as sources of novel antibiotic-producing microorganisms still exist. One such habitat is the rhizosphere of desert shrubs. Here, we show that one desert shrub habitat, the rhizosphere of desert big sagebrush (Artemisia tridentata) is a source of actinomycetes capable of producing an extensive array of antifungal metabolites. Culturable microbial populations from both the sagebrush rhizosphere and nearby bulk soils from three different sites were enumerated and compared, using traditional plate-count techniques and antibiotic activity bioassays. There were no statistical differences between the relative numbers of culturable non-actinomycete eubacteria, actinomycetes and fungi in the rhizosphere versus bulk soils, but PCR amplification of the 16S rRNA gene sequences of the total soil DNA and denaturing gradient gel electrophoresis showed that the community structure was different between the rhizosphere and the bulk soils. A high percentage of actinomycetes produced antimicrobials; and the percentage of active producers was significantly higher among the rhizosphere isolates, as compared with the bulk soil isolates. Also, the rhizosphere strains were more active in the production of antifungal compounds than antibacterial compounds. 16S rRNA gene sequence analysis showed that sagebrush rhizospheres contained a variety of Streptomyces species possessing broad spectrum antifungal activity. Scanning electron microscopy studies of sagebrush root colonization by one of the novel sagebrush rhizosphere isolates, Streptomyces sp. strain RG, showed that it aggressively colonized young sagebrush roots, whereas another plant rhizosphere-colonizing strain, S. lydicus WYEC108, not originally isolated from sagebrush, was a poor colonizer of the roots of this plant, as were two other Streptomyces isolates from forest soil. These results support the hypothesis that the rhizosphere of desert big sagebrush is a promising source of habitat-adapted actinomycetes, producing antifungal antibiotics.     

Topographic control of vegetation in a mountain big sagebrush steppe

Mountain big sagebrush steppes in Wyoming have strong spatial patterning associated with topography. We describe the spatial variability of vegetation in a sagebrush steppe, and test the relationship between topography and vegetation using canonical correlation. Results of the analysis suggest that the main control over vegetation distribution in this system is wind exposure. Exposed sites are characterized by cushion plant communities and Artemisia nova, and less exposed sites by the taller sagebrush species Artemisia tridentata ssp. vaseyana. Topographic depressions and leeward slopes are characterized by aspen stands and nivation hollows. Measurements of soil microclimate suggest that a major influence of topographic position on vegetation is snow redistribution and its effect on soil moisture and temperature.Abbreviations ARNO Artemisia nova - ARTRW Artemisia tridentata ssp. wyomingensis - ARTRV Artemisia tridentata ssp. vaseyana - PUTR Purshia tridentata - RIP riparian community - POTR Populus tremuloides - NIV nivation hollow community     

Antagonistic actinomycetes from Moroccan soil to control the grapevine gray mold

One hundred and forty-two different actinomycete strains were isolated from rhizosphere soil of Vitis vinifera L. sampled from four Moroccan areas. To evaluate the antifungal effect of the different collected actinomycete isolates, five fungi known to be phytopathogens (Pythium ultimum, Fusarium oxyysporum f. sp. albedinis, Sclerotium rolfsii, Verticillium dahliae and Botrytis cinerea) were used. Results showed that 24 isolates had an in vitro inhibitory effect toward at least 4 of the indicator fungi, but only 9 inhibited all these phytopathogens. These nine isolates were subsequently evaluated individually using in vitro grapevine plantlets for their ability to protect against plant gray mold. We demonstrate here that pre-inoculation of plantlets with these isolates allow them to withstand Botrytis cinerea. Six of these strains were shown to belong to the genus Streptomyces and three to the genus Micromonospora. These findings indicate the potential of developing effective actinomycetes from Moroccan habitats for the biological control of Botrytis cinerea. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Isolation of Bacterial Antagonists of Aspergillus flavus from Almonds

Bacteria were isolated from California almond orchard samples to evaluate their potential antifungal activity against aflatoxin-producing Aspergillus flavus. Fungal populations from the same samples were examined to determine the incidence of aflatoxigenic Aspergillus species. Antagonistic activities of the isolated bacterial strains were screened against a nonaflatoxigenic nor mutant of A. flavus, which accumulates the pigmented aflatoxin precursor norsolorinic acid (NOR) under conditions conducive to aflatoxin production. Using solid and liquid media in coculture assays, 171 bacteria isolated from almond flowers, immature nut fruits, and mature nut fruits showed inhibition of A. flavus growth and/or inhibition of NOR accumulation. Bacterial isolates were further characterized for production of extracellular enzymes capable of hydrolyzing chitin or yeast cell walls. Molecular and physiological identification of the bacterial strains indicated that the predominant genera isolated were Bacillus, Pseudomonas, Ralstonia, and Burkholderia, as well as several plant-associated enteric and nonenteric bacteria. A set of 20 isolates was selected for further study based on their species identification, antifungal phenotypes, and extracellular enzyme production. Quantitative assays using these isolates in liquid coculture with a wild-type, aflatoxin-producing A. flavus strain showed that a number of strains completely inhibited fungal growth in three different media. These results indicate the potential for development of bacterial antagonists as biological control agents against aflatoxigenic aspergilli on almonds.     

Actinomycetes isolated from medicinal plant rhizosphere soils: diversity and screening of antifungal compounds,indole-3-acetic acid and siderophore production

A total of 445 actinomycete isolates were obtained from 16 medicinal plant rhizosphere soils. Morphological and chemotaxonomic studies indicated that 89% of the isolates belonged to the genus Streptomyces, 11% were non-Streptomycetes: Actinomadura sp., Microbispora sp., Micromonospora sp., Nocardia sp, Nonomurea sp. and three isolates were unclassified. The highest number and diversity of actinomycetes were isolated from Curcuma mangga rhizosphere soil. Twenty-three Streptomyces isolates showed activity against at least one of the five phytopathogenic fungi: Alternaria brassicicola, Collectotrichum gloeosporioides, Fusarium oxysporum, Penicillium digitatum and Sclerotium rolfsii. Thirty-six actinomycete isolates showed abilities to produce indole-3-acetic acid (IAA) and 75 isolates produced siderophores on chrome azurol S (CAS) agar. Streptomyces CMU-PA101 and Streptomyces CMU-SK126 had high ability to produced antifungal compounds, IAA and siderophores.     

Molecular and phenotypic characterization of potential plant growth-promoting <Emphasis Type="Italic">Pseudomonas</Emphasis> from rice and maize rhizospheres

In this study, Pseudomonas species were isolated from the rhizospheres of two plant hosts: rice (Oryza sativa cultivar Pathum Thani 1) and maize (Zea mays cultivar DK888). The genotypic diversity of isolates was determined on basis of amplified rDNA restriction analysis (ARDRA). This analysis showed that both plant varieties selected for two distinct populations of Pseudomonas. The actual biocontrol and plant promotion abilities of these strains was confirmed by bioassays on fungal (Verticillum sp., Rhizoctonia solani and Fusarium sp.) and bacterial (Ralstonia solanacearum and Bacillus subtilis) plant pathogens, as well as indole-3-acetic acid (IAA) production and carbon source utilization. There was a significant difference between isolates from rice and maize rhizosphere in terms of biological control against R.  solanacearum and B.  subtilis. Interestingly, none of the pseudomonads isolated from maize rhizosphere showed antagonistic activity against R.  solanacearum. This study indicated that the percentage of pseudomonad isolates obtained from rice rhizosphere which showed the ability to produce fluorescent pigments was almost threefold higher than pseudomonad isolates obtained from maize rhizosphere. Furthermore, the biocontrol assay results indicated that pseudomonad isolated from rice showed a higher ability to control bacterial and fungal root pathogens than pseudomonad isolates obtained from maize. This work clearly identified a number of isolates with potential for use as plant growth-promoting and biocontrol agents on rice and maize.     

白马雪山云南黄芪与灰毛康定黄芪根际微生物物种多样性及抗生物膜活性菌株筛选

【背景】细菌生物膜是造成病原菌耐药性增强和持续感染的主要因素,但目前尚无针对抗菌膜的特效药物。特境植物根际微生物可产生大量具有提高宿主免疫功能的活性成分,极具抗生物膜药源开发潜力。【目的】了解滇西北高寒特境白马雪山分布的云南黄芪与灰毛康定黄芪植物根际微生物的物种多样性,并对可培养菌株进行抑菌与抗生物膜活性筛选。【方法】采用宏基因组技术结合传统微生物培养方法,对采自我国云南迪庆藏族自治州德钦县白马雪山的云南黄芪与灰毛康定黄芪的根际微生物进行物种多样性研究,并通过“孔板法”测定其可培养菌株发酵液乙酸乙酯粗浸膏的抗菌、抗生物膜活性。【结果】宏基因组测序结果显示,云南黄芪根际土壤样本中的微生物来自6门7纲8目8科9属10种,其中栖热菌属为优势菌群;灰毛康定黄芪根际土壤样本中的微生物来自6门8纲10目11科14属15种,其中慢生根瘤菌属为优势菌群。通过纯培养共获得145株可培养菌株,包括112株细菌和33株真菌。其中,云南黄芪根际细菌59株,共计16属35种,优势属为假单胞菌属和链霉菌属;根际真菌19株,共计4属5种,优势属为曲霉属;灰毛康定黄芪根际细菌53株,归属于16属29种,优势属为芽孢杆菌属与寡养单胞菌属;根际真菌14株,归属于3属4种,优势属为曲霉属。从不同种水平上选择51株细菌和7株真菌为代表菌株进行抗生素药源评估,发现5株细菌及1株真菌发酵液的乙酸乙酯粗浸膏具有中等至较强的抗革兰阳性菌活性,而且其中4株具有抗MRSA生物膜活性,最终确定了链霉属放线菌Streptomyces fulvissimus KTA1和曲霉属真菌Aspergillus fumigatus YNF5为潜力活性菌株。【结论】首次报道了滇西北地区高寒特境黄芪属植物根际微生物具有较好的物种多样性,而且具有一定的抗生素药用资源开发潜力。本研究对滇西北高寒特境特色植物来源的微生物资源开发利用与保护具有重要的借鉴意义。     

Pathogenicity and specificity of Neozygites tanajoae and Neozygites floridana (Zygomycetes: Entomophthorales) isolates pathogenic to the cassava green mite

The cassava green mite (CGM), Mononychellus tanajoa, a native of South America was accidentally introduced into Africa where it causes serious crop losses. The possibility of introducing classical biological agents from the native home of CGM into Africa was investigated. Thus, we conducted a series of laboratory assays of the native fungal pathogens, Neozygites tanajoae from Brazil and Neozygites floridana from Colombia and Brazil, and compared them with N. tanajoae isolates from Benin. Infectivity of both fungal species, was assayed against the twospotted spider mite, Tetranychus urticae, and against the red mite, Oligonychus gossypii. Pathogenicity against CGM and host range studies were conducted by transferring adult females of each mite species to leaf discs containing sporulated cadavers with a halo of conidia of each fungal isolate. All isolates caused some degree of infectivity to CGM. None of the isolates of N. floridana and N. tanajoae tested were pathogenic to O. gossypii, and only two isolates infected T. urticae. Most isolates from Brazil were highly virulent and infected only CGM. Sixteen N. tanajoae isolates caused more than 89% mortality and more than 62% of the CGM became mummified. A mummified CGM is characteristically a swollen, brown fungus-killed mite that has great potential to produce conidia. However, high mortality was not always associated with high mummification. The median mummification time ranged from 4.4 to 6.7 days. Five Brazilian isolates caused >75% mummification with a median mummification time <5 days. Isolates that cause high mummification in a short period of time would be more likely to cause epizootics and to establish in the new environment. Therefore, these isolates would be the best candidates for introduction to Africa.     

辽宁碱蓬根际土壤真菌多样性的季节变化及其耐盐性

采用稀释平板法和形态学鉴定法进行了辽宁碱蓬(Suaeda liaotungensis)根际土壤真菌的分离鉴定及季节、盐度对真菌多样性的影响分析。结果表明:春、夏、秋、冬四季辽宁碱蓬根际土壤中的真菌菌落数分别为6410、4180、5730和3340,种类分别为6属13种、9属16种、11属31种、6属12种,共分离鉴定出13属42种真菌;其根际土壤真菌的多样性指数、丰富度指数和均匀度指数,从大到小的次序均为秋夏春冬,3种指数均在秋季达到峰值;多样性指数和丰富度指数差异较大,均匀度指数波动较小,春季和冬季的均匀度指数相近。辽宁碱蓬根际土壤真菌种类明显多于无植被土壤,是无植被土壤的7倍,多样性指数和均匀度指数分别是无植被土壤的4—8倍和1—1.2倍;其根际土壤真菌优势种群包括青霉属(Penicillium)、葡萄穗霉属(Stachybotrys)、枝孢属(Cladosporium)、木霉属(Trichoderma)、曲霉属(Aspergillus)和镰孢属(Fusarium);4个季节的优势菌不尽相同,但均有一个共同的优势菌属青霉属。以Na Cl浓度梯度法制造盐胁迫生境,检测辽宁碱蓬根际土壤真菌对盐胁迫的响应,结果表明:60%左右的菌种能耐受5%以下盐度、15%左右的菌种能耐10%—20%盐度;筛选出了6株高度耐盐菌:细交链孢霉、草酸青霉、产黄青霉、烟曲霉、细极链格孢和赭曲霉,其中最高耐盐菌种赭曲霉可耐20%盐度。创新之处在于以盐度和季节为变量检测辽宁碱蓬根际土壤真菌的种群构成,为其深入研究奠定了理论基础。     

不同生境黑果枸杞根际与非根际土壤微生物群落多样性

研究典型生境黑果枸杞根际与非根际土壤微生物群落多样性及其与土壤理化性质间的关系,为进一步研究黑果枸杞抗逆性提供理论数据。采集新疆精河县艾比湖地区(EB)盐碱地、乌苏市(WS)路旁荒地、五家渠市(WQ)人工林带的黑果枸杞根际与非根际土壤,利用Illumina-MiSeq高通量测序技术分析细菌和真菌群落组成和多样性。结果表明:根际土壤细菌多样性高于非根际土壤(WQ除外),而根际真菌多样性低于非根际土壤。WQ非根际土壤细菌和真菌多样性均高于EB和WS;根际细菌多样性排序为EBWSWQ,根际真菌多样性排序为WSEBWQ。根际土壤优势细菌门依次是变形菌门、拟杆菌门、放线菌门、酸杆菌门,真菌优势门为子囊菌门、担子菌门。根际土壤细菌变形菌门、拟杆菌门、酸杆菌门的相对丰度高于非根际土壤,而厚壁菌在根际土壤中的丰度显著降低,真菌优势门丰度在根际土和非根际土中的变化趋势因地区而异; Haliea、Gp10、Pelagibius、Microbulbifer、假单胞菌属、Thioprofundum、Deferrisoma是根际土壤细菌优势属;多孢子菌属、支顶孢属、Corollospora、Cochlonema是根际真菌优势属。细菌、真菌优势类群(门、属)的组成以及丰富度存在地区间差异,厚壁菌门在EB地区的丰富度显著高于含盐量较低的WS、WQ;盐碱生境EB中根际土壤嗜盐细菌的丰度高于非盐碱生境(WQ、WS),如盐单胞菌属、动性球菌属、Geminicoccu、Pelagibius、Gracilimonas、Salinimicrobium等。小囊菌属是EB根际真菌的最优势属,Melanoleuca是WQ和WS的最优势属,地孔菌属、Xenobotrytis、Brachyconidiellopsis、多孢子菌属等在EB根际土壤中的丰度显著高于WQ和WS。非盐碱生境(WS和WQ)的微生物群落之间的相似性较高,并且高于与盐碱环境(EB)之间的相似性,表明土壤含盐量对微生物群落组成丰度具有重要的影响。     

Investigation of actinomycete diversity in the tropical rainforests of Singapore

Five thousand actinomycetes were isolated from soil samples collected from rainforests in Singapore and the generic identities of these isolates were determined by using a procedure that combined morphological, chemotaxonomic and 16S rDNA sequence-based phylogenetic analyses. Actinomycetes belonging to a total of 36 genera were identified. The most abundant isolates are members of Streptomyces, Micromonospora, Actinoplanes, Actinomadura, Nonomuria, Nocardia and Streptosporangium. By phylogenetic analysis of 16S rDNA sequences of our isolates together with those of known actinomycete species, we also evaluated the species diversity of several genera including Streptomyces, Micromonospora, Nonomuria, and Actinomadura. We found that: first, the tropical isolates are present in most clades represented by known species; and second, many tropical isolates form new clades distant from the known species, indicating the presence of unidentified taxa at both species and genus levels. Based on these results, we conclude that actinomycete diversity in the tropical rainforest is very great and should represent an excellent source for discovery of novel bioactive compounds. Received 17 March 1999/ Accepted in revised form 24 June 1999     

Growth stimulation in bean (Phaseolus vulgaris L.) by Trichoderma

Trichoderma species are commonly used as biological control agents against phytopathogenic fungi and some strains are able to produce metabolites that enhance plant growth. In the current study we evaluated the production of potential growth-promoting metabolites, rhizosphere competence and endophytism for 101 isolates of Trichoderma from Colombia, and assessed the relationship of these factors to the enhancement of early stages of growth on bean seedlings. Twenty percent of these Trichoderma strains were able to produce soluble forms of phosphate from phosphoric rock. Only 8% of the assessed strains showed consistent ability to produce siderophores to convert ferric iron to soluble forms by chelation. Sixty percent of isolates produced indole-3-acetic acid (IAA) or auxin analogues. The production of any of these metabolites was a characteristic of specific strains, as the ability to produce these metabolites varied greatly within species. Moreover, the production of these substances did not correlate with enhanced growth on bean seedlings, measured as the combined increase in length of roots and aerial parts in the V3 stage of growth. Seven Trichoderma isolates significantly improved the growth of bean seedlings. However, metabolite production varied widely in these seven strains, and some isolates did not produce any of the assessed growth-promoting metabolites. Results indicated that growth was enhanced in the presence of rhizosphere competent and endophytic strains of Trichoderma, and these characteristics were strain-specific and not characteristic for species.     

Variable responses of insects to hybrid versus parental sagebrush in common gardens

Both ecological and genetic mechanisms have been proposed to explain patterns of herbivore attack on interspecific plant hybrids, but distinguishing among them can be difficult in natural hybrid zones. We performed a common-garden experiment to evaluate four genetic hypotheses: dominance, additivity, elevated hybrid susceptibility, and elevated hybrid resistance. Censuses and cage experiments were used to compare insect responses to basin big sagebrush (Artemisia tridentata spp. tridentata), mountain big sagebrush (A. t. vaseyana), and their F₂ progeny. After two growing seasons, hybrid shrubs resembled mountain big sagenbrush in size, but were more similar to basin big sagebrush in flower production. Censuses of naturally colonizing insects (the gall midge Rhopalomyia obovata, the bagworm moth Apterona helix, and the aphid Obtusicauda coweni) tended to support the dominance hypothesis: if the insect clearly discriminated between the two parents, its frequency on hybrids closely resembled that on one of the parents. Moreover, colonization of hybrids in all three cases suggested a dominance deviation toward the susceptible parent rather than toward the resistant parent. In contrast to the censuses, cage experiments involving two insects supported the hybrid-susceptibility hypothesis; both survival and growth of the grasshopper Melanoplus sanguinipes and growth of the leaf beetle Trirhabda pilosa were higher on hybrid shrubs than on either parent. Because many secondary compounds have been determined to occur at intermediate concentrations in F₂ shrubs, dominance for susceptibility may indicate that insects respond to plant traits (e.g., oviposition stimulants and deterrents) in a threshold manner. Mechanisms underlying increased hybrid susceptibility are less clear, but our experimental design makes environmental explanations (e.g., the plant-stress hypothesis) unlikely. Although we eliminated several confounding factors, our results agree with the conclusion from natural hybrid zones that insect responses to hybrid plants are likely to be idiosyncratic; even congeneric species did not respond similarly to hybrid and parental plants.     

Molecular characterization and evaluation of mycorrhizal capacity of Suillus isolates from Central Spain for the selection of fungal inoculants

Suillus fungal specimens of pine forests from a Mediterranean area of central Spain (Madrid region) were studied based on molecular and physiological analysis of sporocarps to obtain fungal native inocula to produce mycorrhizal Pinus halepensis Miller in nursery. Variation within the internal transcribed spacer (ITS) region of the ribosomal RNA genes of Suillus was examined by restriction fragment length polymorphism (RFLP) and direct sequencing of polymerase chain reaction products. Ribosomal DNA (rDNA) spacers were amplified from pure cultures obtained from fruit bodies of a range of Suillus species: Suillus bellinii (Inzenga) Watling, Suillus bovinus (Pers.) Kuntze, Suillus collinitus (Fr.) Kuntze, Suillus granulatus (L.) Snell, Suillus mediterraneensis (Jacquet. & Blum) Redeuil, Suillus luteus L. (Gray), and Suillus variegatus (Sw.) Kuntze. Interspecific variation in the length and number of restriction sites of the amplified ITS region was observed. This variation was confirmed by sequencing, which allowed us to identify some isolates. This is the first time that the ITS sequence of S. mediterraneensis is completely described. No intraspecific rDNA variation was observed within isolates of S. collinitus, S. mediterraneensis, and S. luteus. The phylogenetic analysis established the close relationship among these Mediterranean fungal species. As a further step to characterize the different isolates and to understand the relation between genetic and functional diversity, some physiological variables were evaluated. Intraspecific variation in axenic fungal growth and in mycorrhizal capacities was detected, especially within S. collinitus isolates. The fungal isolates stimulated the growth of P. halepensis in different rates. These studies indicated that ITS analysis, in conjunction with mycorrhizal tests, provides suitable combined tools for the analysis of Suillus spp. in a small geographic area for selecting isolates with final afforestation purposes.     

Pathogenicity of fungal isolates (Ascomycota: Hypocreales) against Peregrinus maidis, Delphacodes kuscheli (Hemiptera: Delphacidae), and Dalbulus maidis (Hemiptera: Cicadellidae), vectors of corn diseases

Preliminary screening assays were carried out on 17 isolates from five fungal species Beauveria bassiana, Lecanicillium muscarium, Metarhizium anisopliae, Isaria farinosa, and I. fumosorosea. The three most effective isolates against Peregrinus maidis (Hemiptera: Delphacidae) were B. bassiana CEP 147, CEP 150, and CEP 189. There were no consistent differences found in males and females regarding fungal susceptibility. However, more females than males were proportionally infected. There was not a correlation between the percentage of conidial germination and the percentage of mortality caused by fungal infection in any of the treatments. Only B. bassiana CEP 147, which caused a cumulative mortality of 69.8 ± 6.4% after 7 days post-inoculation, was selected to be assayed against adults of P. maidis, Delphacodes kuscheli (Hemiptera: Delphacidae), and Dalbulus maidis (Hemiptera: Cicadellidae). In pathogenicity tests significant differences were observed among treatments. After 2 weeks post-inoculation, both D. kuscheli (cumulative mortality of 73.3 ± 9.0%) and P. maidis (cumulative mortality of 68.6 ± 6.7%) were significantly more susceptible than D. maidis (cumulative mortality of 49.9 ± 9.7%) to the selected isolate.     

Actinomycetes and fungi isolated from plant-parasitic nematode infested soils: screening of the effective biocontrol potential,indole-3-acetic acid and siderophore production

Root-knot nematodes are serious pathogens that severe damage to major crops. They damage plant root system that caused significant yield losses. Moreover, the predisposition of nematode-infected plants is secondary infection from fungal plant pathogen that additional adverse effects on plant growth. Our target is to find the antagonist for control nematode, and secondary infection agents and stimulate plant growth. Twenty-three plant-parasitic nematode infested soils were taken from some provinces in the northern and center of Thailand and actinomycetes and fungi were isolated. Eighty-three isolates belong to actinomycete and 67 isolates were fungi. The predominant actinomycete taxa was Streptomyces (97.6%). The predominant fungal taxa were Penicillium (37.3%) and Fusarium (32.8%). All actinomycete and fungal isolates were subjected for primary screening in vitro for their effects on egg hatching and juvenile mortality of Meloidogyne incognita. Secondary screening was evaluated for antagonist effect on plant pathogenic fungi collected from nematode-infected plant, plant growth hormone (indole-3-acetic acid; IAA) and siderophore production. From primary screening, 7 actinomycete and 10 fungal isolates reduced egg hatching and kill juveniles of M. incognita after 7 days incubation. In secondary screening, 10 nematophagous microbes produced IAA and 9 isolates produced hydroxamate siderophore. Streptomyces sp. CMU-MH021 was selected as a potential biocontrol agent. It reduced egg hatching rate to 33.1% and increased juvenile mortality rate to 82% as contrasted to the control of 79.6 and 3.6%, respectively. This strain had high activity to against tested fungi and high ability on IAA (28.5 μg ml⁻¹) and siderophore (26.0 μg ml⁻¹) production.     

一株抗茶炭疽菌和魔芋镰刀菌的淀粉酶产色链霉菌的分离、鉴定及生防潜能

【背景】放线菌是一类重要的生防菌,具有强大的代谢活性,能产生抗生素、酶、酶抑制剂和激素等天然产物抑制病原物生长。【目的】从茶树根际分离得到放线菌,研究候选放线菌对茶炭疽菌和魔芋镰刀菌的抑菌活性及其生防潜能。【方法】分别以茶炭疽病致病菌Colletotrichum camelliae和魔芋茎腐病致病菌Fusarium solani为指示菌,采用土壤稀释涂布法、平板对峙法和菌丝生长速率法,从茶树根际土壤中分离、筛选拮抗放线菌,并根据菌株的形态特征、生理生化特性和系统发育分析结果对其进行分类鉴定,并开展候选放线菌的产促生相关物质和分泌细胞壁水解酶能力的定性检测试验。【结果】共分离得到14株拮抗放线菌,菌株A-dyzsc04-2的拮抗效果最强,被鉴定为淀粉酶产色链霉菌（Streptomyces diastatochromogenes）。该菌株的活菌体对C. camelliae和F. solani的抑制率分别为66.71%±1.23%和71.59%±2.46%,其无菌发酵滤液对2种指示菌的抑菌率均大于90%;此外,菌株A-dyzsc04-2还具有产嗜铁素和葡聚糖酶以及溶解无机磷的能力。【结论】菌株A-dyzsc04-2是一株优良的生防菌,具有较高的开发利用价值,研究结果为菌株A-dyzsc04-2防治茶炭疽病和魔芋茎腐病提供了理论支撑。     

川蔓藻内生及根际细菌多样性与抑菌活性研究

该研究采用稀释涂布法结合形态观察、16S rRNA基因序列分析,对广西北海川蔓藻(Ruppia maritima)内生及根际细菌的物种多样性进行了研究,并采用琼脂扩散法和光度计法分析了其粗提物抑制马尔尼菲青霉菌活性。结果表明:从川蔓藻中分离到可培养内生细菌26株,根际可培养细菌31株。分别将内生细菌归属为10科12属13种,根际分离出细菌归属为9科14属19种,其中5株根际细菌可能为潜在新种。获得8株细菌对马尔尼菲青霉菌有抑制活性,总阳性率为25.0%。其中,菌株BGMRC 2015、BGMRC 2059、BGMRC 2043的粗提物表现出较强的抑制马尔尼菲青霉菌效果,其MIC分别为(1.800±0.045)、(1.881±0.061)、(1.604±0.021)mg·m L~(-1)。川蔓藻中可培养细菌具有较高的物种多样性,蕴藏着丰富的新物种资源,且富含抑菌活性良好的菌株。     

高寒特境中甸黄芪植物根际微生物物种多样性及其抗生物膜活性成分

滇西北高寒地区分布着丰富的黄芪属植物资源，该属植物“根际效应”明显，其根际微生物极具抗菌药用资源研究价值。【目的】认知滇西北高寒特境中甸黄芪根际微生物的物种多样性，探究其可培养菌株次生代谢产物的化学多样性及抗菌、抗生物膜活性。【方法】采用宏基因组和微生物纯培养方法对中甸黄芪植物根际微生物进行物种多样性分析，同时采用高效液相色谱（high performance liquid chromatography，HPLC）、超高效液相色谱-质谱联用法（ultra-performance liquid chromatography-mass spectrometry，UPLC-MS）结合“微量肉汤稀释法” “孔板法”等多级联合筛选策略综合评估可培养菌株的抗菌活性药源研究价值。【结果】对中甸黄芪根际土壤样本的微生物分类操作单元（operational taxonomic units，OTU）进行分类注释，得到22门54纲105目187科316属856种微生物，其中优势菌群为慢生根瘤菌属。纯培养共获得27属54种95株可培养菌株，包括20属33种54株细菌和7属21种41株真菌，优势属分别为芽孢杆菌属和青霉属。其中，1株细菌Pseudomonas tolaasii ZTB4和3株真菌Aspergillus tabacinus ZNF17、Lecanicillium aphanocladii ZNF15、Umbelopsis nana ZTF31的次生代谢产物具有广谱抗菌活性。同时，菌株ZTB4和ZNF17的次生代谢产物也显示出优秀的抗耐甲氧西林金黄色葡萄球菌（methicillin-resistant Staphylococcus aureus，MRSA）生物膜活性，并已验证这2株菌株的主要活性成分分别为环脂肽类与黄酮类。【结论】中甸黄芪植物根际微生物物种多样性较为丰富，其可培养菌株次生代谢产物有较好的化学多样性和抗菌、抗生物膜活性。研究结果为我国特境特色微生物药用资源的开发利用提供理论依据。     

Nodulation patterns of actinorhizal plants in the family rosaceae

Patterns of nodulation, growth, andFrankia — host specificity have not been well characterized for the actinorhizal genera in the family Rosaceae because of the scarcity ofFrankia isolates from these taxa. Furthermore, the few isolates available from actinorhizal Rosaceae have consistently failed to nodulate plants from the host genus. In a series of experiments, species of rosaceousDryas, Cowania, Cercocarpus, Fallugia, andPurshia were inoculated withFrankia isolates, crushedDryas actinorhizae, and neoglacial soils to ascertain whether any of these inocula would effectively induce nodulation. Neoglacial soils from Alaska and Canada nodulated not only the localDryas drummondii, but alsoCercocarpus betuloides, Cowania mexicana andPurshia tridentata from distant and ecologically diverse locales as well as nonrosaceous, actinorhizal species ofAlnus, Elaeagnus, Myrica, andShepherdia. But of eightFrankia isolates, including two fromPurshia tridentata and one fromCowania mexicana, none were able to induce nodulation onPurshia orCowania species. Globular, actinorhizae-like nodules incapable of acetylene reduction were produced onC. betuloides inoculated withFrankia isolates. Crushed nodule suspensions fromDryas drummondii nodulated rosaceousCowania, Dryas andPurshia, as well as non-rosaceousElaeagnus, Myrica, andShepherdia species. Nodules produced by inoculation ofCowania mexicana andPurshia tridentata with crushed, dried nodule suspensions fromDryas drummondii reduced acetylene to ethylene, indicating nitrogenase activity for these nodulated plants. These data suggest that a similar microsymbiont infects the actinorhizal genera in the family Rosaceae.