植物内生菌的研究进展

植物内生菌是存在于植物内部与植物密不可分的一类微生物,包括内生真菌和内生细菌两大类。随着研究的不断深入,发现内生菌不仅产生多种有益的生物学作用,如防病、促生、固氮等,还能对寄主植物的生长发育产生不利影响。近年来,植物内生菌已成为我国微生物学领域研究的热点问题之一。本文综述了植物内生真菌和内生细菌的研究概况,以期更好地研究和了解植物内生菌。     

禾草内生真菌生物碱的研究进展

综述了国内外近20a以来在禾草内生真菌生物碱方面的研究进展。目前,已发现至少4大类10余种生物碱与内生真菌有关。各类生物碱中典型代表物的分子结构已完全清楚,部分内生真菌在离体条件下可产生除黑麦草碱外的生物碱,但产碱量较其在植物体中所产生的低很多,可相差150余倍之多。随着生物技术的发展,美国、新西兰等国在波胺、麦角碱和loline的生物合成途径方面已有了初步的进展,对个别具有重要功能基因以及其所编码的酶已有了深入研究。各种生物碱的致毒机理尚未完全清楚,除饱和吡咯化合物为新陈代谢类毒素和神经性毒素外,其余3类生物碱均为神经性毒素。生物碱可增加禾草对40余种害虫的抗性,并可增加对某些线虫和病害的抗性。诸多因素均可影响寄主植株中生物碱的种类和浓度,包括寄主植株：种群和生态型,植物品种和基因型,植株的部位和生长期;环境：气候因素,土壤养分,季节和年度变化;内生真菌菌株和草地管理利用方式等。用于生物碱检测的主要方法为预分离检测法和直接检测法,其中高效液相色谱法以其分离能力强、选择性高、测定灵敏度高,操作简单,可在室温下进行,应用范围极广的优点而广泛应用。目前,国际关于禾草内生真菌生物碱研究的重点包括创造不含对家畜有毒素的有益禾草-内生真菌共生体,开展基因工程研究以及合理利用生物碱,使其成为新一代的“生物农药”。     

一种大连细叶早熟禾内生真菌的形态学和系统发育学研究

【目的】鉴定分离自辽宁省大连市细叶早熟禾中的内生真菌。【方法】在辽宁省大连市的黄金山和白玉山采集细叶早熟禾(Poa awgws啡L.),通过显微镜观察植物是否含菌,统计样品中内生真菌的检出率;从含菌样品中分离内生真菌,纯化后在马铃薯葡萄糖琼脂(PDA)培养基上培养3周,测定分离菌株的形态学特征;运用IQS法提取分离菌株的总DNA,利用特异性引物妒增分离菌株的P-Tubulin(tubB)和Translation elongation factor 1-a(碎4)基因片段,利用DNAssist 2.2和ClustalX 1.81软件进行序列的多重比对和分析,运行PAUP 4 beta 10、MrModeltest 2.3选择合适的模型,用MrBayes 3.1.2构建最大似然树,分析分离菌株的系统发育学特征。【结果】共釆集植物样品262个,其中内生真菌的检出率达57.3%;根据分离地点和菌落特征筛选和纯化了10个分离菌株,选用其中6个分离菌株进行形态学特征的调查,发现分离菌株呈典型的Neotyphodium属形态学特征,进而和其他宿主为早熟禾属植物的内生真菌进行比较,发现分离菌株的特征与Epichlo谷liyangensis相近,但分生抱子较小[(5.3±0.3)|jmx(2.9±0.2)Mm],生长速度稍慢(每周8.7±1.4 mm);基于5个分离菌株tubB和tefA片段序列的系统发育学特征显示,它们的如65和对^序列分别为单拷贝,并与五.中的第二拷贝聚为一类。【结论】分离菌株在原始宿主植物上没有形成子座,分离菌株的形态学特征和系统发育学特征均与五./zyawge/ts/j?有明显差异。兼顾其它早熟禾属植物内生真菌的宿主植物的种类和地理分布特征,这些菌株有可能属于一个新的内生真菌类群。     

Fungal endophyte diversity from tropical forage grass Brachiaria

ABSTRACT

Background

Microbial endophytes can be beneficial to host plants, improving resistance against biotic and abiotic stress. While beneficial fungal endophytes have been well characterised in temperate grasses, knowledge about fungal endophytes of tropical grasses is limited.     

Lignicolous freshwater ascomycota from Thailand: 1.Ascotaiwania sawada and its anamorph stateMonotosporella

Single ascospore isolates ofAscotaiwania sawada gave rise to conidia of aMonotosporella species. Repeated isolations confirmed that theMonotosporella was the anamorph ofA. sawada. These findings are discussed and compared with the other known anamorph-teleomorph stages of freshwater fungi. This species ofMonotosporella is new to science but because it has not been found in nature it is referred to as theMonotosporella state ofA. sawada.     

醉马草内生菌的分离、鉴定及杀虫效果

【目的】探明醉马草内生菌的种类,筛选对农作物虫害有毒杀作用的菌株。【方法】采用研磨法从健康醉马草植物的根、茎、叶和种子中进行菌种分离;通过对其形态、培养特征、生理生化及其他生物学特性的研究;16SrDNA及ITS序列的系统发育学分析进行鉴定;用玻片浸渍法和喷雾法筛选产杀虫活性物质菌株。【结果】获得细菌89株,分别属于枯草芽孢杆菌属(Bacillus)、链霉菌属(Streptomyces)、棒状杆菌属(Corynebacterium)、叶杆菌属(Phyllobacterium)、鞘脂单胞菌属(Sphingomonnas)、类芽孢杆菌(Paenibacillus)、假单胞菌属(Pseudomonas)和不动杆菌属(Acinetobacter)8个属;真菌2株,分别属于麦角菌属(Claviceps)和毛壳菌属(Chaetomium)。经初筛及复筛,内生菌娄彻氏链霉菌Streptomyces rochei(GA)和黑麦麦角菌Claviceps purpurea(PF-2)发酵液粗提物对棉蚜虫(Aphis gossypii)致死率达85%以上。【结论】醉马草内生菌株PF-2和GA的粗提代谢物对棉蚜虫有明显的毒杀作用,为开发新的生物源农药提供了生物源物质。     

Neoplaether, a new cytotoxic and antifungal endophyte metabolite from Neoplaconema napellum IFB-E016

A new diphenyl ether, named neoplaether, together with five known compounds monomethylsulochrin, physcion, helvolic acid, ergosterol and ergosterol peroxide, was isolated from the culture of Neoplaconema napellum IFB-E016, an endophytic fungus residing in the healthy leaves of Hopea hainanensis. The structure of neoplaether was elucidated by a correlative interpretation of its infrared spectrometry, mass spectroscopy and nuclear magnetic resonance spectra, and confirmed by its single-crystal X-ray diffraction analysis. Neoplaether exhibited significant cytotoxic activity against the human nasopharyngeal epidermoid tumor KB cell line, with an IC(50) value of 13.0 microg mL(-1), comparable to that of 5-fluorouracil (2.5 microg mL(-1)) co-assayed as a positive reference. In addition, it showed antifungal activity against Candida albicans, with a minimal inhibitory concentration value of 6.2 microg mL(-1) (amphotericin as a positive control had a value of 1.5 microg mL(-1)).     

The genusBrachydesmiella from submerged wood in the tropics, including a new species and a new combination

During studies of fungi occurring on wood submerged in freshwater streams in the tropics, five species ofBrachydesmiella, namely,B. anthostomelloidea, B. biseptata, B. caudata, B. orientalis comb. nov., andB. verrucosa sp. nov., were collected. There are described and illustrated from natural substrata, wooden test blocks, and from pure culture, based mainly on collections made in Thailand.Brachydesmiella verrucosa differs from the other four species in having 3-celled, coarsely verrucose, ampoule-shaped conidia, with a long, thin, cylindrical, verruculose apical cell. A key toBrachydesmiella species is provided.     

云南红豆杉内生真菌Talarom yces sp.T1BF的化学成分研究

采用色谱技术从云南红豆杉内生真菌Talaromyces sp.T1BF的PDA平板发酵物中分离得到4个化合物.经波谱数据分析及与文献对照,分别鉴定为5α,6α-环氧-24( R)-甲基胆甾-7,22-二烯-3β-醇(1)、醌茜素(2)、苔色酸(3)和间甲基苯酚(4).化合物1为首次从该属真菌中分离获得.     

植物内生菌研究进展及其存在的问题

植物内生菌是一个多样性十分丰富的微生物类群 ,分布于没有外在感染症状的健康植物组织内 ,并与宿主植物协同进化 ,其存在和作用长期以来一直为人们所忽视。随着研究领域的不断拓宽和研究方法的不断深入 ,植物内生菌的生态和生理作用及其作为潜在的生防资源和外源基因载体 ,在农业和医药领域中的巨大应用潜力 ,已逐渐成为国内外研究的热点。本文综述了近 10年来国内外植物内生菌研究的概况和最新进展 ,并对该研究领域中存在的若干问题进行了探讨。     

醉马草内生菌的分离、鉴定及杀虫研究

摘要：【目的】探明醉马草内生菌的种类,筛选对农作物虫害有毒杀作用的菌株。【方法】采用研磨法从健康醉马草植物的根、茎、叶和种子中进行菌种分离;通过对其形态、培养特征、生理生化及其他生物学特性的研究;16S rDNA及ITS序列的系统发育学分析进行鉴定;用玻片浸渍法和喷雾法筛选产杀虫活性物质菌株。【结果】获得细菌89株,分别属于枯草芽孢杆菌属（Bacillus）、链霉菌属(Streptomyces)、棒状杆菌属（Corynebacterium）、叶杆菌属（Phyllobacterium）、鞘脂单胞菌属（Sph     

Book review

Summary

A new coprophilous species of Coniochaeta, C. burtii, with 512-spored asci, is described from deer dung from Glen Dochart, Perthshire.     

植物内生菌抗虫工程研究进展

生物防治已成为植物虫害防治的发展方向,越来越多地受到人们的关注。利用植物内生菌进行抗虫性的研究不断深入。本着重论述了内生菌的抗虫机制及其影响因素。同时,对抗虫策略的实施也进行了探讨。     

Cocoicola livistonicola, sp. nov., and notes onCocoicola cylindrospora from palms

Cocoicola livistonicola, a new species from senescent petioles ofLivistona chinensis in Hong Kong, is described. It differs from previously reportedCocoicola species in having differently shaped ascospores. Notes onCocoicola cylindrospora which was recently collected onCocos nucifera in the Seychelles are also provided.     

内生真菌感染对黑麦草抗盐性的影响

以感染内生真菌的多年生黑麦草（Lolium perenne L．）（SR4000）为实验材料,建植内生真菌感染（EI）和不感染（EF）的黑麦草种群,并对其进行盐胁迫实验,通过观察生长和生理生态指标的变化,分析内生真菌对宿主植物抗盐性的影响。结果表明,内生真菌感染对宿主黑麦草的营养生长没有增益效应,相反在高盐浓度下,EI种群的分蘖能力和地上部分生物量均低于EF种群;但内生真菌能够改变宿主种群生物量的分配格局,将更大比例的生物量分配于根系。在高盐浓度下,内生真菌感染可导致黑麦草叶内的脯氨酸含量显著增加、可溶性糖含量显著降低,但对PSⅡ光化学效率Fv/Fm值的变化没有影响。总体来看,内生真菌感染并未改善宿主黑麦草的抗盐性。     

内生真菌感染对宿主植物高羊茅锌耐受性的影响

以感染内生真菌(Neotyphodium coenophialum)和未感染内生真菌的高羊茅(Festuca arundinacea Schreb.)为实验材料,在营养液中加入ZnSO4进行锌胁迫实验,分析内生真菌对宿主植物锌耐受性的影响。与未感染内生真菌的植株相比,内生真菌感染对高羊茅的总生物量没有显著增益作用,但增加了分蘖数和叶片延伸生长累积值。内生真菌感染降低了高羊茅中Zn2+的总含量,改变了锌在高羊茅中的分配,增加叶鞘中锌的含量,减少叶片中锌的含量。在高锌浓度下,内生真菌感染对净光合速率的变化没有影响,但是显著提高了其宿主的PSⅡ光化学效率(Fv/Fm)。总体来看,内生真菌感染改善宿主高羊茅的锌耐受性。     

Investigation into the ability of Gluconacetobacter sacchari to live as an endophyte in sugarcane

The relatively low numbers and sporadic pattern of incidence of the acetic acid bacterium Gluconacetobacter sacchari with the pink sugarcane mealybug (PSMB) Saccharicoccus sacchariCockerell (Homoptera: Pseudococcidae) over time and from different sugarcane-growing regions do not indicate that Glac. sacchari is a significant commensal of the PSMB, as has been previously proposed. This study was conducted to investigate the hypothesis that Glac. sacchari is, like its closest relative Glac. diazotrophicus, an endophyte of sugarcane (Saccharum officinarum L.). In this study, bothGlac. sacchari and Glac. diazotrophicus were isolated from internal sugarcane tissue, although the detection of both species was sporadic in all sugarcane-growing regions of Queensland tested. To confirm the ability of Glac. sacchari to live endophytically, an experiment was conducted in which the roots of micropropagated sugarcane plantlets were inoculated with Glac. sacchari, and the plantlets were subsequently examined for the presence of the bacterium in the stem cells. Pure cultures of Glac. sacchari were grown from homogenized surface sterilized sugarcane stems inoculated withGlac. sacchari.Electron microscopy was used to provide further conclusive evidence that Glac. sacchari lives as an endophyte in sugarcane. Scanning electron microscopy of (SEM) sugarcane plantlet stems revealed rod-shaped cells of Glac. sacchari within a transverse section of the plantlet stem cells. The numbers of bacterial cells inside the plant cell indicated a successful infection and colonization of the plant tissue. Using transmission electron microscopy, (TEM) bacterial cells were more difficult to find, due to their spatial separation. In our study, bacteria were mostly found singularly, or in groups of up to four cells inside intercellular spaces, although bacterial cells were occasionally found inside other cells.