新疆哈密地区盐湖放线菌的多样性及其功能酶的筛选

[目的]本研究旨在了解新疆哈密地区盐湖放线菌多样性及产功能酶的特性.[方法]采用含有5%与10%NaC1的4种分离培养基,利用稀释平板涂布法对盐湖土壤样品进行分离;通过形态特征、耐盐性实验、抑菌实验及16S rRNA基因测序的基础上进行系统发育学分析;利用五种筛选培养基定性检测放线菌的产酶活性.[结果]从盐湖土壤样品中分离到63株放线菌,其中中等嗜盐放线菌47株;抑菌活性实验结果表明:23株放线菌对痢疾杆菌和/或其它病原菌有抑菌活性;功能酶筛选结果表明:3株放线菌产蛋白酶、46株产淀粉酶、14株产酯酶、34株产半乳糖苷酶、5株产纤维素酶.16S rRNA基因的系统发育学分析结果表明盐湖放线菌类群比较丰富.[结论]新疆哈密地区盐湖放线菌资源丰富,产酶特性良好,为开发利用极端环境微生物资源奠定了基础.     

塔克拉玛干沙漠腹地胡杨林土壤细菌多样性分析

【目的】对塔克拉玛干沙漠腹地胡杨林土壤细菌多样性进行初步探索,为下一步从中筛选可用于生物饲料或生物肥料的微生物奠定基础。【方法】采用可培养方法,进行细菌的分离纯化。对各菌株进行革兰氏染色及淀粉酶、酯酶、纤维素酶和NaCl耐受浓度的测定,并提取各菌株基因组DNA,进行16SrRNA基因扩增、测序及系统进化树的绘制,分析其多样性。【结果】共分离得到27株菌,其中放线菌门(Actinobacteria)16株,变形菌门(Proteobacteria)4株,厚壁菌门(Firmicutes)6株,拟杆菌门(Bacteroidetes)1株。革兰氏染色结果表明,5株菌为革兰氏阴性,其余为革兰氏阳性;酶活测定结果表明,15株菌具有淀粉酶活性,9株菌具有酯酶活性,9株菌具有纤维素酶活性;NaCl耐受浓度测定结果显示,NaCl浓度为2%时所有菌株均能生长,5%时能生长的有22株,15%时能生长的有1株。【结论】塔克拉玛干沙漠腹地胡杨林土壤中存在较丰富的细菌类群,且具有一定的酶学活性和NaCl耐受性,具有进一步研究开发的价值。     

津巴布韦烟叶中淀粉酶和蛋白酶产生菌的分离及鉴定

目的:从津巴布韦烟叶中分离产蛋白酶菌和产淀粉酶能力最高的菌株,并对其进行鉴定。方法:采用淀粉富集培养基和酪蛋白富集培养基分别分离津巴布韦烟叶中的产淀粉酶和产蛋白酶菌株,通过生理生化实验和16SrRNA序列分析鉴定分离的菌株。结果:产蛋白酶菌株菌体不透明、表面有褶皱,蛋白酶酶活为52.10±0.13 U/mL;产淀粉酶菌株菌体表面呈黏状,淀粉酶酶活为3.69±0.07 U/mL;产蛋白酶与产淀粉酶的2株菌均与枯草芽孢杆菌的16S rRNA序列有100%的相似性,结合生理生化指标初步鉴定为枯草芽孢杆菌。结论:获得的2株菌在降解烟叶的蛋白质和淀粉过程中可能起重要作用。     

Rope-producing strains of Bacillus spp. from wheat bread and strategy for their control by lactic acid bacteria

Two types of white wheat bread (high- and low-type loaves) were investigated for rope spoilage. Thirty of the 56 breads tested developed rope spoilage within 5 days; the high-type loaves were affected by rope spoilage more than the low-type loaves. Sixty-one Bacillus strains were isolated from ropy breads and were characterized on the basis of their phenotypic and genotypic traits. All of the isolates were identified as Bacillus subtilis by biochemical tests, but molecular assays (randomly amplified polymorphic DNA PCR assay, denaturing gradient gel electrophoresis analysis, and sequencing of the V3 region of 16S ribosomal DNA) revealed greater Bacillus species variety in ropy breads. In fact, besides strains of B. subtilis, Bacillus licheniformis, Bacillus cereus, and isolates of Bacillus clausii and Bacillus firmus were also identified. All of the ropy Bacillus isolates exhibited amylase activity, whereas only 32.4% of these isolates were able to produce ropiness in bread slices after treatment at 96 degrees C for 10 min. Strains of lactic acid bacteria previously isolated from sourdough were first selected for antirope activity on bread slices and then used as starters for bread-making experiments. Prevention of growth of approximately 10(4) rope-producing B. subtilis G1 spores per cm(2) on bread slices for more than 15 days was observed when heat-treated cultures of Lactobacillus plantarum E5 and Leuconostoc mesenteroides A27 were added. Growth of B. subtilis G1 occurred after 7 days in breads started with Saccharomyces cerevisiae T22, L. plantarum E5, and L. mesenteroides A27.     

Isolation and characterization of antagonistic Bacillus subtilis strains from the avocado rhizoplane displaying biocontrol activity

AIM: This study was undertaken to isolate Bacillus subtilis strains with biological activity against soil-borne phytopathogenic fungi from the avocado rhizoplane. METHODS AND RESULTS: A collection of 905 bacterial isolates obtained from the rhizoplane of healthy avocado trees, contains 277 gram-positive isolates. From these gram-positive isolates, four strains, PCL1605, PCL1608, PCL1610 and PCL1612, identified as B. subtilis, were selected on the basis of their antifungal activity against diverse soil-borne phytopathogenic fungi. Analysis of the antifungal compounds involved in their antagonistic activity showed that these strains produced hydrolytic enzymes such as glucanases or proteases and the antibiotic lipopeptides surfactin, fengycin, and/or iturin A. In biocontrol trials using the pathosystems tomato/Fusarium oxysporum f.sp. radicis-lycopersici and avocado/Rosellinia necatrix, two B. subtilis strains, PCL1608 and PCL1612, both producing iturin A, exhibited the highest biocontrol and colonization capabilities. CONCLUSIONS: Diverse antagonistic B. subtilis strains isolated from healthy avocado rhizoplanes have shown promising biocontrol abilities, which are closely linked with the production of antifungal lipopeptides and good colonization aptitudes. SIGNIFICANCE AND IMPACT OF THE STUDY: This is one of the few reports dealing with isolation and characterization of B. subtilis strains with biocontrol activity against the common soil-borne phytopathogenic fungi F. oxysporum f.sp. radicis-lycopersici and R. necatrix.     

西双版纳保护区植物根际细菌的筛选及其促生能力研究

【背景】西双版纳保护区具有丰富的生物多样性,而该区域植物根际细菌特别是放线菌及其促生能力相关报道较少。【目的】从西双版纳保护区根际土壤中筛选出植物根际促生菌,并检测其促生能力。【方法】采用5种不同培养基筛选出植物根际促生菌并通过16S rDNA序列分析进行分类学鉴定,运用Salkowski法测定菌株产IAA的能力,CAS法测定菌株产铁载体能力,钼锑抗显色法测定菌株的解磷能力,CMC-Na法测定菌株产纤维素酶能力和改良的Young法测定产淀粉酶能力,综合评价所得菌株的促生能力。【结果】从土样中分离纯化得到14株典型促生菌,经鉴定分别归属于链霉菌属(Streptomyces)、诺卡菌属(Nocardi)、杆菌属(Bacillus)、中华根瘤菌属(Ensifer)、中慢生根瘤菌属(Mesorhizobium)、固氮螺菌属(Azospirillum)和狭单胞菌属(Stenotrophomonas)。其中菌株B433产吲哚乙酸的能力在培养12 d时达到最大值9.23 mg/L;菌株B351、B453、B546这3株菌株产铁载体的能力较强,其Su80%,最高可达86.67%,强度为+++++;菌株B541的解磷能力最强,磷酸根的浓度达到9.79 mg/L;菌株B442综合产纤维素酶能力最强为31.86 U/mL;菌株B412淀粉酶活力为16.07 U/mL。【结论】西双版纳保护区植物根际土壤促生细菌种类丰富,且具有较强的广谱促生能力,有潜在的开发价值,本研究可为此地的微生物资源开发提供可靠的菌株资源依据。     

Burkholderia cepacia XXVI siderophore with biocontrol capacity against Colletotrichum gloeosporioides

Colletotrichum gloeosporioides is the causal agent of anthracnose in mango. Burkholderia cepacia XXVI, isolated from mango rhizosphere and identified by 16S rDNA sequencing as a member of B. cepacia complex, was more effective than 6 other mango rhizosphere bacteria in inhibiting the model mango pathogen, C. gloeosporioides ATCC MYA 456. Biocontrol of this pathogen was demonstrated on Petri-dishes containing PDA by > 90 % reduction of surface colonization. The nature of the biocontrol metabolite(s) was characterized via a variety of tests. The inhibition was almost exclusively due to production of agar-diffusible, not volatile, metabolite(s). The diffusible metabolite(s) underwent thermal degradation at 70 and 121 °C (1 atm). Tests for indole acetic acid production and lytic enzyme activities (cellulase, glucanase and chitinase) by B. cepacia XXVI were negative, indicating that these metabolites were not involved in the biocontrol effect. Based on halo formation and growth inhibition of the pathogen on the diagnostic medium, CAS-agar, as well as colorimetric tests we surmised that strain XXVI produced a hydroxamate siderophore involved in the biocontrol effect observed. The minimal inhibitory concentration test showed that 0.64 μg ml(-1) of siderophore (Deferoxamine mesylate salt-equivalent) was sufficient to achieve 91.1 % inhibition of the pathogen growth on Petri-dishes containing PDA. The biocontrol capacity against C. gloeosporioides ATCC MYA 456 correlated directly with the siderophore production by B. cepacia XXVI: the highest concentration of siderophore production in PDB on day 7, 1.7 μg ml(-1) (Deferoxamine mesylate salt-equivalent), promoted a pathogen growth inhibition of 94.9 %. The growth of 5 additional strains of C. gloeosporioides (isolated from mango "Ataulfo" orchards located in the municipality of Chahuites, State of Oaxaca in Mexico) was also inhibited when confronted with B. cepacia XXVI. Results indicate that B. cepacia XXVI or its siderophore have the potential to be used as a biological control agent against C. gloeosporioides; thus diminishing environmental problems caused by the current practices to control this disease.     

Genotypic and phenotypic differentiation of an antifungal biocontrol strain belonging to Bacillus subtilis

Physiological and molecular fingerprints of biotechnologically relevant rhizobacteria are necessary for registration, patenting, recognition and quality checking of the strains. To characterize the biological control agent, Bacillus subtilis B2g, the strain was compared with other plant-associated B. subtilis isolates. Phenotypic characterization included biochemical and nutritional properties, in vitro activity and analysis of potential antagonistic mechanisms towards several plant pathogenic fungi. According to the phenotypic characteristics, it was not possible to differentiate the biocontrol agent from the other strains, although the enzymatic fingerprint was unique. Genotypic diversity among the isolates was characterized by molecular fingerprinting methods using REP-PCR (repetitive extragenomic palindromic PCR), and macrorestriction of genomic DNA and electrophoretic separation of DNA fragments by pulsed-field gel electrophoresis (PFGE). A protocol for PFGE analysis using restriction enzyme SfiI for B. subtilis was developed. PFGE typing of B. subtilis B2g resulted in a unique fingerprint. Therefore, it was possible to differentiate B. subtilis B2g, the biocontrol agent of Phytovit, from other antifungal B. subtilis isolates.     

Biological control of Sclerotinia sclerotiorum (Lib.) de Bary, the causal agent of white mold, by Pseudomonas species on canola petals

Sclerotinia sclerotiorum is an important pathogen on canola. Due to the public concern over pesticide use, alternative methods of disease control, such as biological control, should be considered. Several bacterial strains were isolated from canola and soja plants. Inhibition of S. sclerotiorum by bacterial strains in vitro was assayed on PDA medium in dual culture test. Eight Pseudomonas sp. strains (PB-3, PB-4, PB-5, PB-6, PB-7, PB-8, PB-10 and PB-11) caused inhibition zone against 5. sclerotiorum hyphal growth. The biocontrol potential of the bacteria was tested in a plant assay. Disease suppression was investigated using a petal inoculation technique. Canola petals were pretreated with bacteria, and then inoculated with 5. sclerotiorum ascospores 24 h later. Greenhouse experiment showed that application of Pseudomonas sp. strains (1 x 10(8) cfu ml(-1)) effectively suppressed S. sclerotiorum (1 x 10(5) ascospores ml(-1)) on petals and all of them achieved significant (P<0.01) disease suppression. Fourteen days after inoculation, strain PB-3 had 88/7% disease control and strain PB-4 had 69/9% disease control. Result from all studies indicates PB-3 to be effective biocontrol against S. sclerotiorum of canola. PB-3, PB-4, PB-7, PB-8, PB-10 and PB-11 were identified as Pseudomonas fluorescens biovar III. PB-5 and PB-6 was identified as Pseudomonas fluorescens biovar II. Strains PB-3, PB-4, PB-6, PB-10 and PB-11 produced protease and HCN. Strain PB-5 produce protease; no HCN.     

In vitro degradation of wheat straw by anaerobic fungi from small ruminants

Anaerobic ruminal fungi may play an active role in fibre degradation as evidenced by the production of different fibrolytic enzymes in culture filtrate. In the present study, 16 anaerobic fungal strains were isolated from ruminal and faecal samples of sheep and goats. Based on their morphological characteristics they were identified as species of Anaeromyces, Orpinomyces, Piromyces and Neocallimastix. Isolated Neocallimastix sp. from goat rumen showed a maximum activity of CMCase (47.9 mIU ml(-1)) and filter paper cellulase (48.3 mIU ml(-1)), while Anaeromyces sp. from sheep rumen showed a maximum xylanolytic activity (48.3 mIU ml(-1)). The cellobiase activity for all the isolates ranged from 178.0-182.7 mIU ml(-1). Based on the enzymatic activities, isolated Anaeromyces sp. from sheep rumen and Neocallimastix sp. from goat rumen were selected for their potential of in vitro fibre degradation. The highest in vitro digestibility of NDF (23.2%) and DM (34.4%) was shown for Neocallimastix sp. from goat rumen, as compared to the digestibility of NDF and DM in the control group of 17.5 and 25.0%, respectively.     

Application of endophytic bacteria for controlling anthracnose disease (Colletotrichum lindemuthianum) on bean plants

Over 100 endophytic bacterial isolates were isolated from surface-sterilised roots of the Fabaceae family in East Azerbaijan farms. These isolates were screened for their in vitro biocontrol activity against Colletotrichum lindemuthianum by dual culture technique using potato dextrose agar (PDA) medium. Eight bacterial isolates (Bacillus subtilis subsp. subtilis, Bacillus atrophaeus, B. tequilensis, B. subtilis subsp. spizizenii, Streptomyces cyaneofuscatus, S. flavofuscus, S. parvus, S. acrimycini) showed promising inhibition on mycelial growth of C. lindemuthianum , and thus, these isolates were selected for greenhouse experiments. The disease control rate using these selected endophytic bacteria was varied from 40 to 76.80% in greenhouse without any negative effects on different growth performance, suggesting that these selected endophytic bacteria are potential to be developed as biocontrol agents.     

Cloning and expression of an endocellulase gene from a novel streptomycete isolated from an East African soda lake

Alkaline cellulase-producing actinomycete strains were isolated from mud samples collected from East African soda lakes. The strains were identified as novel Streptomyces spp. by 16S rDNA sequence analysis. A cellulase gene (cel12A) from Streptomyces sp. strain 11AG8 was cloned by expression screening of a genomic DNA library in Escherichia coli. From the nucleotide sequence of a 1.5-kb DNA fragment, an open reading frame of 1,113 nucleotides was identified encoding a protein of 371 amino acids. From computer analysis of the sequence, it was deduced that the Cel12A mature enzyme is a protein of 340 amino acids. The protein contained a catalytic domain, a glycine-rich linker region, and a cellulose-binding domain of 221, 12, and 107 amino acids, respectively. FASTA analysis of the catalytic domain of Cel12A classified the enzyme as a family 12 endoglucanase and the cellulose-binding domain as a family IIa CBD. Streptomyces rochei EglS was determined as nearest neighbor with a similarity of 75.2% and 61.0% to the catalytic domain and the cellulose-binding domain, respectively. The cell2A gene was subcloned in a Bacillus high-expression vector carrying the Bacillus amyloliquefaciens amylase regulatory sequences, and the construct was transformed to a Bacillus subtilis host strain. Crude enzyme preparations were obtained by ultrafiltration of cultures of the Bacillus subtilis recombinant strain containing the 11AG8 cell2A gene. The enzyme showed carboxymethylcellulase (CMCase) activities over a broad pH range (5-10) with an optimum activity at pH 8 and 50 degrees C. The enzyme retained more than 95% of its activity after incubation for 30 min under these conditions.     

Unrelatedness of Bacillus amyloliquefaciens and Bacillus subtilis

Eight strains of highly amylolytic, sporeforming bacilli (hereafter referred to as Bacillus amyloliquefaciens) were compared with respect to their taxonomic relationship to B. subtilis. The physiological-biochemical properties of these two groups of organisms showed that B. amyloliquefaciens differed from B. subtilis by their ability to grow in 10% NaCl, characteristic growth on potato plugs, increased production of alpha-amylase, and their ability to ferment lactose with the production of acid. The base compositions of the deoxyribonucleic acid (DNA) of the B. subtilis strains consistently fell in the range of 41.5 to 43.5% guanine + cytosine (G + C), whereas that of the B. amyloliquefaciens strains was in the 43.5 to 44.9% G + C range. Hybrid formation between B. subtilis W23 and B. amyloliquefaciens F DNA revealed only a 14.7 to 15.4% DNA homology between the two species. Transducing phage, SP-10, was able to propagate on B. subtilis W23 and B. amyloliquefaciens N, and would transduce B. subtilis 168 (indole(-)) and B. amyloliquefaciens N-10 (arginine(-)) to prototrophy with a frequency of 3.9 x 10(-4) and 2.4 x 10(-5) transductants per plaque-forming unit, respectively. Attempts to transduce between the two species were unsuccessful. These data show that Bacillus amyloliquefaciens is a valid species and should not be classified as a strain or variety of B. subtilis.     

Construction of novel Ruminococcus albus, strains with improved cellulase activity by cloning of Streptomyces rochei endoglucanase gene

The 6.3 kb plasmid pCRB1 containing the eglS gene from Streptomyces rochei, coding for a -1,4 glucanase, was constructed in Bacillus subtilis by using the plasmid vector pIL253 with subsequent activity of the cellulase activity. Strictly anaerobic cellulolytic and xylanolytic strains of Ruminococcus albus, isolated from the rumen of cows and water buffaloes, were used for transformation experiments. The plasmid pCRB1 was introduced by means of electroporation into five freshly isolated strains of R. albus, with frequencies ranging from 10 to 10 /mg of plasmid DNA. Northern analysis demonstrated the expression of the eglS gene in R. albus. All the strains harbouring the heterologous cellulase gene showed an increase of the secreted cellulase activity.     

益生枯草芽胞杆菌B15的性能检测及培养条件研究

对筛选得到的枯草芽胞杆菌B15进行了产酶及拮抗性能的检测,结果表明该菌株能够分泌淀粉酶、蛋白酶、纤维素酶,同时对常见病原体有一定的拮抗作用,具有作为益生菌进行发酵和研制微生态制剂的潜力。采用单因子试验,对B15菌株的培养液组成（碳源、氮源）及培养条件进行了研究,结果表明：以1.5%葡萄糖及0.5%酵母浸出物制成的培养液,在35℃、初始pH6.5左右,装液量为20 ml/250 ml,接种量2%的条件下培养B15,可将活菌数目从原始的1.15×109CFU/ml提高至1.61×109CFU/ml。     

Seed Bacterization with Bacillus subtilis AF 1 Increases Phenylalanine Ammonia-lyase and Reduces the Incidence of Fusarial Wilt in Pigeonpea

Cell free culture filtrate of a biocontrol PGPR strain of Bacillus subtilis showed a concentration-dependent growth and conidiation inhibition of Fusarium udum . the incitant of pigeonpea wilt. In vitro interaction of cell free filtrate of AF 1 induced bulb-like structures in the hyphae of F. udum . Seed treatment with B. subtilis significantly reduced the incidence of pigeonpea wilt, but not with the cell free culture filtrate of AF 1. Seeds bacterized with B. subtilis AF 1 showed an increase in phenylalanine ammonia-lyase (PAL) from day 1, while the peroxidase activity increased from day 6. The results are discussed in terms of induction of host plant resistance in biological control of plant pathogens.     

Evaluation of potential biocontrol rhizobacteria from different host plants of Verticillium dahliae Kleb

AIMS: A screening approach was developed to assess the potential of rhizobacterial strains to control Verticillium wilt caused by Verticillium dahliae Kleb. METHODS AND RESULTS: Sixty randomly chosen antagonistic bacterial strains originally isolated from rhizosphere of three different host plants of V. dahliae--strawberry, potato and oilseed rape--were evaluated for biocontrol and plant growth promotion by analysing in vitro antagonism towards V. dahliae and other plant pathogenic fungi, production of fungal cell wall-degrading enzymes and plant growth-promoting effects on strawberry seedlings. To test the plant growth-promoting effect, a microplate assay with strawberry seedlings was developed. Although the rhizobacterial strains were isolated from different plants they showed effects on the growth of strawberry seedlings. According to the in vitro biocontrol and plant growth-promoting activity, the three best candidates Pseudomonas putida B E2 (strawberry rhizosphere), Ps. chlororaphis K15 (potato rhizosphere) and Serratia plymuthica R12 (oilseed rape rhizosphere) were selected for greenhouse experiments to verify the in vitro screening results. Under greenhouse conditions the isolates selected according to this strategy were as effective, or more effective than commercial biocontrol agents and may therefore possibly be valuable as antagonists of V. dahliae. CONCLUSIONS: In this study, the screening strategy resulted in a selection of three interesting biocontrol candidates against Verticillium: Ps. putida B E2 (strawberry rhizosphere), Ps. chlororaphis K15 (potato rhizosphere) and Ser. plymuthica R12 (oilseed rape rhizosphere). SIGNIFICANCE AND IMPACT OF THE STUDY: A new combination of in vitro screening methods including a microplate assay with strawberry seedlings to test the plant growth promoting effect which allow to more efficiently select potential biological control agents was developed successfully.     

Hydrolytic activities of anaerobic fungi from wild blue bull (Boselaphus tragocamelus)

The anaerobic fungi play an active role in the plant fibre degradation by producing a wide array of potential hydrolytic enzymes in the rumen. In present study, 12 anaerobic fungal strains were isolated from the faecal samples of wild blue bull, and identified as species of Piromyces, Anaeromyces, Orpinomyces and Neocallimastix based on their morphological characteristics. Isolate WNG-12 (Piromyces sp.), showed maximum filter paper cellulase (23 mIU ml(-1)) and xylanase (127 mIU ml(-1)) activity, while WNG-5 (Piromyces sp.) showed maximum carboxymethyl cellulase activity (231 mIU ml(-1)). Based on the results obtained, it can be stated that Piromyces sp. WNG-12 may be a promising isolate in utilizing fibre rich diets in the rumen as evidenced by the production of hydrolytic enzymes in vitro.     

Strategy to select and assess antagonistic bacteria for biological control of Rhizoctonia solani Kühn

A screening strategy was developed to assess the potential of plant-associated bacteria to control diseases caused by Rhizoctonia solani Kühn. About 434 already characterized antagonistic bacterial strains isolated from diverse plant species and microenvironments were evaluated for biocontrol and plant growth promotion by a hierarchical combination of assays. Analyzing in vitro antagonism towards different Rhizoctonia isolates resulted in a selection of 20 potential biocontrol agents. The strains were characterized by their antagonistic mechanisms in vitro as well as their production of the plant growth hormone indole-3-acetic acid. The plant growth promoting effect by antagonistic bacteria was determined using a microtiter plate assay on the basis of lettuce seedlings. Lettuce and sugar beet as host plant were included in the biocontrol experiments in which the antagonistic effect of 17 bacterial isolates could be confirmed in vivo. Sequencing of the 16S rDNA gene and (or) fatty acid methyl ester gas chromatography was used to identify the antagonistic isolates. Molecular fingerprints of isolates obtained by BOX-polymerase chain reaction were compared to avoid further investigation with genetically very similar strains and to obtain unique molecular fingerprints for quality control and patent licensing. According to our strategy, an assessment scheme was developed and four interesting biological control agents, Pseudomonas reactans B3, Pseudomonas fluorescens B1, Serratia plymuthica B4, and Serratia odorifera B6, were found. While S. plymuthica B4 was the best candidate to biologically control Rhizoctonia in lettuce, P. reactans B3 was the best candidate to suppress the pathogen in sugar beet.     

黔西南薏苡内生真菌多样性及其抗菌活性研究

为进一步了解贵州黔西南薏苡(Coix lacryma-Jobi)可培养内生真菌的多样性组成及其抗菌活性。通过组织块分离法,选取薏苡根、叶和种仁为分离对象,进行内生真菌分离,并通过形态学观察,分子生物学特征对所分离内生真菌进行鉴定。采用平板菌块对峙法对薏苡内生真菌进行抗菌活性筛选。从薏苡根、叶和种仁中共分离纯化获得76株薏苡内生真菌,根据形态和分子生物学特征,它们属于10个目、14个科、26个属和1个不明属以及未知菌;以大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphyloccocus aureus)、枯草芽胞杆菌(Bacillus subtilis)、酿酒酵母(Saccharomyces cerevisiae)为指示菌株,对薏苡内生真菌进行抗菌活性筛选。其中有25株对大肠杆菌具有抗菌活性,占分离菌株的比例为32. 9%;有34株对金黄色葡萄球菌具有抗菌活性,其比例为44. 7%;有24株对枯草芽胞杆菌具有抗菌活性,比例是31. 6%;有23株对酿酒酵母具有抗菌活性,占比例为30. 3%;对4种标准菌株都有抗菌活性的有13株,占总株数的17. 1%。薏苡的不同组织结构中存在丰富的内生真菌资源,部分内生真菌具有抑制其他微生物生长的活性,具有产生天然活性产物成分的潜力,为工业应用奠定了资源基础,具有进一步发掘和研究的价值。