量天尺根部内生真菌的多样性

为了解不同量天尺（火龙果）品种根部内生真菌菌群组成及多样性,采集GHL-1、GHL-2、GHL-3、ML-1和DL 5个量天尺品种健康根部样品,进行内生真菌分离,采用形态观察和ITS序列分析相结合的方法进行鉴定、归类。共分离得到内生真菌菌株117株,总体分离率为25.71%,分别隶属于13个属,其中Trichoderma、Fusarium、Chaetomium和Phoma为量天尺内生真菌的优势种群,分别占总菌株数的24.79%、35.04%、10.26%和10.26%;不同量天尺品种内生真菌的结构和组成存在一定差异,GHL-2、GHL-3和DL 3个品种中分离频率最高的内生真菌类群为Fusarium,GHL-1和ML-1分离频率最高的类群为Trichoderma;多样性分析结果反映出不同量天尺品种内生真菌菌群的多样性指数、丰富度指数和均匀度指数水平存在差异,其中GHL-2的3项指数均为最高。表明品种差异对内生真菌的组成和多样性均有影响。     

The ability of the biological control agent Bacillus subtilis, strain BB, to colonise vegetable brassicas endophytically following seed inoculation

The ability of Bacillus subtilis, strain BB, to colonise cabbage seedlings endophytically was examined following seed inoculation. Strain BB was recovered from different plant parts including leaves (cotyledons), stem (hypocotyl) and roots. While high bacterial populations persisted in the roots and lower stem, they were lower in the upper stem and leaves through time. In addition to cabbage, strain BB colonised endophytically the roots of 5 other vegetable brassicas. Fatty acid methyl ester (FAME) and PCR fingerprinting analysis confirmed the reliability of the detection method. Studies conducted with transmission electron microscope (TEM) showed that BB mainly colonised intercellular spaces of cortical tissues including intercellular spaces close to the conducting elements of roots and stem of cabbage seedlings. Gold labelling was specifically associated with BB and the fibrillar material filling the intercellular spaces where bacterial cells were found.     

一株耐盐碱乳酪短杆菌G20响应盐碱胁迫的差异代谢物分析

【目的】探究耐盐碱乳酪短杆菌G20响应盐碱胁迫的代谢物组成以及代谢物合成潜力,为潜在功能分子和盐碱诱导的快速稳定响应逻辑门基因线路的挖掘提供参考。【方法】利用液相色谱-质谱联用技术(LC-MS)检测乳酪短杆菌G20盐碱环境与正常环境下4个生长时期的代谢产物。着重对富含高差异变化倍数代谢物的适应期与指数期进行分析。【结果】乳酪短杆菌G20可以在pH 10.0、9%NaCl环境中正常生长,同时环境pH值会随菌株生长逐步下降。综合正负离子2种模式,乳酪短杆菌G20在盐碱环境下各生长时期间差异代谢物数量分别为正常环境的0.69、0.75和0.81倍。盐碱胁迫诱导下适应期与指数期差异代谢物主要为苯环型化合物、有机酸及其衍生物与有机杂环类化合物。其中上调的有机酸化合物吲哚-3-乙酸、犬尿酸和葡萄糖酸指数期质谱信号强度低于适应期。菌株中可能存在的渗透保护剂有L-瓜氨酸、L-脯氨酸、N-乙酰鸟氨酸和左旋肉碱等。适应期变化倍数较大或质谱信号强度较高的差异化合物有毛果芸香碱、植物鞘氨醇和柠檬酸等,指数期有组胺、L-脯氨酸和硫胺素等。菌株差异代谢通路集中在氨基酸代谢与碳水化合物代谢。菌株代谢物中存在甜菜碱和...     

青藏高原垫状点地梅叶际及内生可培养微生物多样性

【背景】垫状点地梅作为青藏高原最具代表性的垫状植物,其叶际和内生微生物对适应极端环境有重要意义,同时也是一种独特的资源。【目的】探究垫状点地梅叶际和叶内可培养微生物多样性,以及不同生存状态个体之间的微生物差异。【方法】采用纯培养方法分离和纯化3个不同地区垫状点地梅叶际和叶内的细菌、酵母菌和丝状真菌,并用16S rRNA基因和ITS区域序列进行分析鉴定。【结果】最终得到叶际微生物350株,鉴定为22属49种,优势种为Penicillium sajarovii;内生微生物274株,鉴定为19属45种,优势种为Bacillusmycoides;两者的优势属均为Penicillium。垫状点地梅叶际和叶内之间及不同生存状态个体之间微生物的α多样性大多无显著差异,各群落间的成员也有重叠,但物种组成存在显著的空间异质性。【结论】垫状点地梅叶际和叶内有着丰富的可培养微生物资源,来源于不同生存状态的个体或不同部位的微生物物种组成差别较大,微生物对不同环境的选择偏好形成了不同的群落模式。但这些不同来源的微生物群落之间同样存在高比例的共有菌株,这些共有菌株的异养方式和生态位并不固定,可兼共生和腐生生存,生...     

非洲哈茨木霉LMNS-M9的鉴定、生物学特性及其对藜麦的促生作用

【背景】藜麦作为风靡全球的功能性食品,其种子内生非洲哈茨木霉的研究尚未见报道。【目的】从藜麦种子中筛选出有抑菌活性的内生真菌,分析其抑菌和促生作用。【方法】通过形态学及ITS、tef1和rpb2基因序列等方法对筛选出的内生真菌进行鉴定;采用平板对峙和平板倒扣等方法测定内生真菌的抑菌活性;利用盆栽试验测定内生菌株对藜麦生长的影响。【结果】综合形态学特征和系统发育分析结果,确定菌株LMNS-M9为非洲哈茨木霉(Trichoderma afroharzianum)。30℃、pH 5.0和镁离子适宜菌株LMNS-M9菌丝生长和产孢;葡萄糖、乳糖、蛋白胨、磷酸二氢钾适宜菌株LMNS-M9菌丝生长;葡萄糖、硝酸铵、磷酸氢二钾适宜菌株LMNS-M9产孢。菌株LMNS-M9对Botrytis cinerea、Ascochyta caulina、Fusarium citri、Alternaria alternata和Trichothecium roseum的抑制率分别为12.53%、51.96%、52.38%、59.25%和62.04%,挥发物的抑制率分别为35.86%、61.54%、33.33%、41...     

光枝勾儿茶内生真菌多样性及抑菌活性研究

为探索贵州苗药光枝勾儿茶内生真菌类群特征、分布部位及其抑菌活性,该研究采用传统方法对贵州省贵阳市和黔西市光枝勾儿茶内生真菌进行分离,并基于分子生物学及统计学对其分类地位进行鉴定及多样性评价,最后通过微量肉汤倍比稀释法筛选具有抑菌活性的菌株。结果表明:(1)从光枝勾儿茶中分离到191 株内生真菌,隶属于3 个门5 个纲10 个目15 个科19 个属,优势属为叶点霉属(Phyllosticta)、间座壳属(Diaporthe)、葡萄座腔菌属(Botryosphaeria)和刺盘孢属(Colletotrichum)。(2)黔西光枝勾儿茶内生真菌香农-维纳多样性指数(H''_Q=2.112)较贵阳(H''_G=1.801)高,索伦森相似性指数Cs_G-Q为0.923,不同组织香农-维纳多样性指数为茎(H''_S=2.004)>根(H''_R=1.764)>叶(H''_L=1.654)>果实(H''_F=1.473),茎和叶内生真菌的索伦森相似性最高(Cs_S-L=0.667)。(3)筛选出的21株内生真菌对供试菌大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)和沙门氏菌(Salmonella enterica)具有抑菌效果,其中Diaporthe sp. QX4G6对大肠杆菌、金黄色葡萄球菌和沙门氏菌的最小抑菌浓度分别为12.5、6.25、12.5 mg·mL^-1,最小杀菌浓度分别为12.5、6.25、12.5 mg·mL^-1。以上研究结果揭示了光枝勾儿茶蕴藏丰富的内生真菌资源,不同地区及组织内生真菌类群组成有差异,多个分离菌株具有抗菌活性,为光枝勾儿茶内生真菌天然抗菌药物或药源研发奠定了基础。     

蛇足石杉内生真菌产黄青霉菌MT-40中xanthocillin类似物生物合成基因簇的挖掘及鉴定

Xanthocillin具有显著的抗菌活性,结构中含有独特的异腈基。本文通过对蛇足石杉(Huperzia serrata)内生真菌产黄青霉菌(Penicillium chrysogenum)MT-40基因组的测序分析,利用本地BLAST等生物信息学分析工具挖掘具有合成xanthocillin类似物潜力的基因簇,结合米曲霉(Aspergillus oryzae)NSAR1异源表达技术实现基因簇中关键基因的功能鉴定。结果成功从内生真菌P.chrysogenum MT-40中发现一个合成xanthocillin类似物的生物合成基因簇(命名为for),for基因簇中的关键生物合成基因forB编码的异腈基合成酶可以催化合成2-formamido-3-(4-hydroxyphenyl)acrylic acid,基因forG编码的P450酶可以催化2-formamido-3-(4-hydroxyphenyl)acrylic acid的二聚化生成xanthocillin类似物N,N′-(1,4-bis(4-hydroxyphenyl)buta-1,3-diene-2,3-diyl)diformamide。本文研究结果为进一步从真菌中发现xanthocillin类似物提供参考。     

Effect of open-air fumigation with sulphur dioxide and ozone on phyllosphere and endophytic fungi of conifer needles

The phyllosphere microbial populations inhabiting the needles of three conifer species, Scots pine (Pinus sylvestris L.), Sitka spruce (Picea sitchensis L.) and Norway spruce (Picea abies (L.) Karst.), exposed to SO₂ and O₃, in an open-air fumigation experiment were analysed over a 3 year period using serial dilution after washing, direct plating and a fluorescein diacetate (FDA) enzyme assay. Total fungal populations ranged from 10² to 10⁵ colonyforming units (CPU) g^?1 fresh weight of needles. The dominant fungi isolated from needles varied with tree species and isolation technique; Aureobasidium pullulans (de Bary) Arnaud was most common on Scots pine and Norway spruce and white yeasts on Sitka spruce using the dilution plating method. However, direct plating of needle segments onto culture media indicated that Sclerophoma pythiophila (Corda) Hohnel was dominant on Scots pine and A. pullulans on Sitka and Norway spruce. Green needles of Sitka spruce were found to be endophytically colonized by Rhizosphaera kalkhoffii Bubak, but seldom by Lophodermium piceae (Fuckel) Hohn during extensive sampling in 1990. Statistical analyses revealed significant differences (P<0.05) between plots in the 3 year mean of the total fungal populations or the fungal biomass (FDA assay) on all three tree species. Differences between plots were also observed for a number of dominant component species. Data were also analysed for treatment effects. A significant effect of SO₂ treatment was observed on the total fungal populations on Sitka spruce (P<0.05) which were reduced markedly by the low-SO₂ treatment, while the O₃ treatment caused a significant increase in total fungal numbers on Scots pine (P<0.05). The FDA activity on needles of both Scots pine and Sitka spruce was noticeably higher in the 0₃-only treatment plot, but the overall O₃ effect was not significant. Treatment effects were also detected on the occurrence of component species. The serial dilution method revealed an SO₂ effect (P<0.05) of a reduction in the occurrence of pink yeasts on Sitka spruce and an O₃ effect (P<0.05) of an increase in the occurrence of S. pythiophila on Sitka spruce (P<0.01) but a decrease of Epicoccum nigrum Link and Cladosporium spp. on Scots pine. The direct-plating method revealed an SO₂ effect of an increase in S. pythiophila on Norway spruce (P<0.05). Ozone treatment caused a significant increase in the isolation of a black strain of A. pullulans on Norway spruce (P<0.05). Endophytic colonization of Sitka spruce needles by R. kalkhoffii was found to be increased on two occasions by O₃ exposure.     

Potential and pitfalls of trying to extend symbiotic interactions of nitrogen-fixing organisms to presently non-nodulated plants,such as rice

It has been a long-standing goal in the field of biological nitrogen fixation to extend nitrogen-fixing symbioses to presently non-nodulated cereal plants, such as rice. A number of researchers have recently described the induction of nodule-like structures on the roots of cereals primarily by rhizobia, in either the presence or absence of plant cell-wall-degrading enzymes or plant hormones. We briefly review this research and discuss the potential problems associated with the introduction of nitrogen-fixing microbes in novel physiological environments, such as rice roots. The results of experiments carried out in China on the induction of nodule-like structures on rice roots by rhizobia are highlighted. In addition, we present preliminary results of a series of experiments designed to repeat and evaluate these results using a variety of microscopic techniques and molecular genetic approaches.     

Temporal relationships betweenZea mays,ostrinia nubilalis (Lep.: Pyralidae) and endophyticBeauveria bassiana

The entomopathogenic fungus,Beauveria bassiana (Balsamo) Vuillemin, was applied to whorl-stage (V7) corn,Zea mays L., by foliar application of a granular formulation of corn grits containing conidia or by injection of a conidial suspension. All plants were infested with European corn borer larvae,Ostrinia nubilalis (Hübner), at the V7 (whorl), V12 (late-whorl), or V17 (pretassel) stage of plant development. Plants infested at whorl and late-whorl stages had significantly more European corn borer tunneling than did plants infested at the pretassel stage. The percentage of plants colonized byB. bassiana did not differ significantly among the whorl, late-whorl, and pretassel stages. As the plants matured,B. bassiana was isolated from different plant areas, with the pith more frequently colonized than the leaf collars. Foliar application ofB. bassiana provided immediate suppression ofO. nubilalis in those plants infested at whorl stage. The reduced efficacy ofB. bassiana at the intermediate plant stages relative to efficacy at harvest is discussed. The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.