中国腐霉属的水生种类

腐霉属是一个世界广布分类群,迄今为止已报道约100种.它们主要分布于土坡中,条件适合时侵染植物引起病害,造成多种经济植物的重大损失.腐霉也见水中,它们既可寄生于藻类及一些小动物上,也可腐生于动植物残骸上.在研究微生物与水质关系时,腐霉的作用是不可忽视的.在发酵工业上,腐霉可望用于生产多种酶类及转化街族化合物,所以它是一个经济意义较重要的类群.     

中国大陆的腐霉属菌物

综述了中国大陆腐霉属的研究进展,内容包括腐霉属菌物的发生、分布、分类鉴定、致病性、所致植物病害防治及腐霉的利用等方面.至今,中国已报道的腐霉属菌物有64个种,其中有13个种作为世界新种进行了描述,这13个新种分别为:顶生腐霉Pythium acrogynum,孤雌腐霉P.amasculinum,百色腐霉P.baisense,北方腐霉P.boreale,短枝腐霉P.breve,壁合腐霉P.connatum,镰雄腐霉P.falciforme,贵阳腐霉P.guiyangense,广西腐霉P.guangxiense,下雄腐霉P.hypoandrum,昆明腐霉P.kummingense,南宁腐霉尸nanningense和中国腐霉P.sinensis.瓜果腐霉Pythium aphanidermatum为优势种,在全国普遍引起多种植物严重的猝倒病和根、茎、叶、果腐烂病.其中,大多数腐霉种对植物具有致病性,44个种寄生植物,1个种:紫菜腐霉P.porphyrae寄生红藻和紫菜,2个种:卡地腐霉P.carolinianum 和贵阳腐霉P.guiyangense寄生蚊子幼虫,2个种:努氏腐霉P.nunn和寡雄腐霉P.oligandrum 是真菌的重寄生菌.相比之下,台湾报道的腐霉种有48种(其中新种1个:四季腐霉P.sukuiense),海南报道的腐霉种有28种.对中国大陆腐霉属的研究前景也进行了讨论.     

卵菌的系统学和分类学

引言卵菌Oomycetes生于水中、土内和陆上;淡水比海水中多,静水比流水中多;陆生多为植物寄生菌。腐生、寄生、专性寄生和共生均有。寄生于藻类、蕨类、种子植物、昆虫、鱼类及其他真菌上,引起病害。如水霉危害鱼苗和鱼卵,绵霉引起烂秧,疫霉和腐霉摧毁多种栽培植物,霜霉和白锈为害各种经济植物,病害流行时常造成严重的经     

水霉菌总DNA提取方法研究

本实验采用EP管反复冻融和研磨的破壁方式,利用溶菌酶法、CTAB法、改良CTAB法、尿素法、十二烷基硫酸钠(SDS)法等5种方法,分别对5种鱼类致病性水霉菌(寄生水霉、多子水霉、异株水霉及两未定种)的基因组DNA进行提取,并采用紫外分光光度计和ITS区基因(包括5.8S rDNA)PCR扩增对DNA进行了评价.紫外分光光度计检测结果表明,5种方法均可提取到水霉菌DNA,其中改良CTAB法提取的5种水霉菌的DNA产量和质量最高,A260/A280在1.79-1.82之间,浓度为45μg/mL;PCR检测结果表明,只有改良CTAB法提取的DNA全部扩增到明亮、整齐、无拖尾的特异性条带,其他几种方法均存在暗带或无带现象.因此,改良CTAB法可以作为水霉菌DNA提取以开展分子生物学研究的首选方法.     

中国单轴霉分类的研究——Ⅰ.寄生在唇形科和苋科植物上的新种

本文描述了单轴霉属(Plasmopara)霜霉菌的二个新种。它们是:寄生于唇形科植物香薷(Elsholtzia ciliata(Thunb.)Hyland)上的香薷单轴霉(Plasmopara elsholtziae spnov.)及苋科植物牛膝(Achyranthes bidentata Bl.)上的牛膝单轴霉(Plasmopara achyranthis sp. nov.)。牛膝单轴霉是本属真菌寄生于苋科植物的首次记录。上述二新种都有拉丁文及汉文描述,还分别讨论了新种与其同科或不同科寄主上的近似种之间的区别。     

农杆菌介导的携带egfp基因载体转化寡雄腐霉

【目的】寡雄腐霉(Pythium oligandrum Drechsler)是一种对动、植物和环境无害,兼具杀菌和增产效果的生防真菌。通过研究建立农杆菌介导的寡雄腐霉遗传转化体系。【方法】选用EHA105、AGL-1、LBA4404三种农杆菌菌株对寡雄腐霉进行遗传转化研究,通过对影响遗传转化效果的条件参数试验优化,确立适宜寡雄腐霉遗传转化的农杆菌菌株及转化条件,建立农杆菌介导的寡雄腐霉遗传转化体系。【结果】经研究发现,所选3种农杆菌菌株中EHA105菌株对寡雄腐霉的遗传转化效果最好,其次是AGL-1菌株,LBA4404菌株转化效果不好。EHA105菌株经IM(含300μmol/L AS)诱导培养至OD_(600)=0.6时,与浓度为10~6–10~7个/m L的寡雄腐霉孢子悬浮液以1–10:1的比例混合,在25–26°C以液体振荡的方式避光共培养72 h(pH 5.0,含300μmol/L AS),寡雄腐霉菌体液体振荡恢复培养24 h,涂布抗性选择平板筛选寡雄腐霉转化子,即可得到寡雄腐霉基因工程菌株,其转化率可达到130个转化子/106个孢子。【结论】本研究首次构建了农杆菌介导的寡雄腐霉遗传转化体系,研究结果可为寡雄腐霉的生防机制及分子育种研究提供技术支撑。     

应用RAPD技术鉴定地霉的实验研究

目的探讨RAPD技术在快速鉴定地霉中应用。方法用E.Z.N.A.yeastDNAkit提取地霉菌基因组DNA,采用随机引物AP3(5'-TCGTAGCCAA-3')、ATG(5'-ATGGATCGGC-3')、RP2(5'-AAGGATCAGA-3')、OPA-10(5'-GTGATCGCAG-3')对临床上致病性白地霉、林生地霉皮损株和血液株的基因组DNA进行扩增,对各病原菌的DNA指纹的特征进行分析。结果成功提取了地霉的基因组DNA,其纯度和浓度均能满足PCR反应的要求。利用4种引物对基因组DNA进行扩增,不同种真菌的DNA显示不同的DNA带型,分离自不同感染部位的同种不同株真菌的DNA显示的主要DNA带型基本一致。结论采用E.Z.N.A.yeastDNAkit提取的地霉基因组DNA可以用于PCR反应。RAPD法鉴定地霉菌简单、快速、特异,可用于临床诊断。     

中国单轴霉分类的研究Ⅱ.寄生在伞形科上的新种和已知种

本文报道在我国伞形科植物上寄生的4种单轴霉(Plasmopara)。其中寄生于鸭儿芹(Cryptotaenia japonic Hassk)上的鸭儿芹单轴霉(Plasmopara cryptotaeniae sp.nov.)和寄生于水芹[Oenanthe javanica(BI.)DC.]及卵叶水芹(O.rosthornii Diels)上的水芹单轴霉(Plasmopara oenantheae sp.nov.)是2个新种。寄生于变豆菜(Sanicula chinensis Bunge)上的变豆菜单轴霉(Plasmopara saniculae Traian et O.savulescu)是亚洲的新记录种。     

腐霉属三个中国新记录种(英文)

通过对分离自中国土壤中的腐霉菌进行形态特征和DNA序列分析,发现3个中国新记录种:钟形腐霉Pythium campanulatum、卵突腐霉P. oopapillum和多卵腐霉 P. plurisporium。根据采集材料对其进行了形态学特征描述和图解,研究标本保存于中国科学院微生物研究所菌物标本馆(HMAS)。     

中国霜霉的几个新种

本文描述了霜霉属(Peronospora)两个新种。即:寄生于唇形科植物香薷(Elsholtziapatrnia Garcke)上的香薷霜霉(Peronospora elsholtziae sp.nov.)和寄生于大花益母草(Leonurusmacranthus Maxim.)上的大花益母草霜霉(Peronospora leonuri sp.nov);单轴霉属(Plasmopara)一个新种,寄生于车前草科植物车前草(Plantago major L.)上的车前草单轴霉(Plasmoparaplantaginicola sp.nov.,);盘梗霉属(Bremia)一个新种,寄生于石竹科植物莫石竹(Moehringialateriflora L.)上的莫石竹盘梗霉(Bremia moehringiae sp.nov.).这四个新种均有拉丁文和中文描述。还分别讨论了新种与同科近似种之间的区别。     

Species-specific PCR primers for Pythium developed from ribosomal ITS1 region

AIMS: To develop a specific method for distinguishing and detecting Pythium species. METHODS AND RESULTS: Twenty PCR primers were designed from the sequences of the rDNA internal transcribed spacer 1 (ITS1) region from 34 Pythium species. The specificity of these forward primers paired with ITS2 or ITS4 and reverse universal primers was tested. Five species-specific primers were obtained, other primers showed different specificity to Pythium species. The specific amplifications enabled nine Pythium species to be differentiated. Specific detection of Pythium aphanidermatum from infested plants and P. dimorphum from soil was demonstrated. CONCLUSIONS: A method for identifying nine Pythium species using specific PCR amplification was achieved. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of its rapidness and ease, the results of PCR amplified with different primers can be a powerful method for identifying Pythium species and detecting or monitoring the target fungus directly from plant material, soil and water samples.     

Development of Multiplex PCR to Detect Five Pythium Species Related to Turfgrass Diseases

The objective of this study was to develop multiplex PCR detection method for five Pythium species associated with turfgrass diseases, Pythium aphanidermatum, Pythium arrhenomanes, Pythium graminicola, Pythium torulosum and Pythium vanterpoolii. Species‐specific primers and two common primers were designed based on the sequences of the internal transcribed spacer region of ribosomal DNA. Another primer set by which all organisms would be amplified in 18S rDNA was used as a positive control. When these total nine primers were applied to the multiplex PCR, all species were individually discriminated in the mixture of five species culture DNA. Furthermore, all five Pythium species were detected in naturally infected plants using the multiplex PCR.     

PYTHIUM SYLVATICUM鉴定及其专一性PCR引物

从杭州采集的水稻、棉花和大豆猝倒苗中分离到国内新记录腐霉种Pythium sylvaticum。该腐霉为异宗配合种,菌丝膨大体球形或柠檬形,雄器异丝生,藏卵器光滑,每个藏卵器上1~3个雄器,雄器常在接近藏卵器处形成二叉状分枝,卵孢子不满器。测定了该种4 个菌株的核糖体内转录间隔区(ITS)的序列,根据与59种腐霉ITS序列的比较,设计了P. sylvaticum种专一性引物PSF1和PSR2。实际结果表明:该引物能从11种共14株腐霉DNA中特异性地扩增P. sylvaticum,从而与其它10种腐霉区分。     

Pythium species associated with cavity spot on carrots in Norway

Carrot roots with cavity spot lesions from eight different counties in Norway were sampled and Pythium species were isolated on selective medium. Pythium spp. were characterised morphologically and by species-specific PCR. Laboratory experiments with inoculations of carrot roots were performed. A total of 130 isolates out of 230 Pythium -like isolates tested with PCR were identified as pathogenic species of Pythium. These were P. intermedium (29%), P. sulcatum (23%), P. sylvaticum (16%), P. violae (15%) and a possible new Pythium species designated P . ' vipa ' (18%). There were some differences between geographical regions and ages of cavities regarding the frequency of the different species isolated. When rating sunken lesions in the laboratory inoculation experiments, P. ' vipa ' was the most aggressive and P. violae the least aggressive species. P. intermedium and P. ' vipa ' caused more discolouration of the infected carrot tissue than the other species. The importance of the different Pythium spp. as agents of cavity spot in Norway is discussed.     

Comparative studies on the DNA of Pythium species and some possibly related taxa

DNA from 14 Pythium species, Verrucalvus flavofaciens and Zoophagus insidians was characterized by CsCl-bisbenzimide density gradients in order to investigate its taxonomic potential. A few incomplete analyses were made for other species. All clearly assignable Pythium species produced three DNA bands in the gradient. Pythium undulatum along with Verrucalvus and Zoophagus produced only two bands. Another possible exception, which needs further investigation, is P. vexans. The DNA had a relatively constant banding pattern in CsCl gradients. The small number (eight) of DNA criteria that were available were subjected to cluster analysis to assess the relationships between replicates and species. This restricted database, similar in size to the number of criteria used in morphological taxonomy, provided an independent assessment of the values that have been attached to generic and subgeneric classifications. This approach enabled assessments to be made of relationships between species that have incomplete life-histories and which therefore lack features essential for traditional taxonomic decisions.     

腐霉属分类性状评价及其中国的种

本文对腐霉属Pythium Pringsheim的研究历史作了简单的回顾,对该属的分类性状和系统进行了论述和评价,最后对中国已发现的55种腐霉,以检索表的方式进行了分类、检索。     

Pythium ornacarpum: a new species with ornamented oogonia isolated from soil in France

Pythium ornacarpum sp. nov. was isolated from a soil sample taken from Genlis in the Burgundy region of France. This species is unique because of its ornamented oogonia which are completely surrounded by antheridial filaments. The fungus is closely related to Pythium echinulatum Matthews. Morphological and reproductive aspects of this species as well as a study by PCR of the sequence of the internal transcribed spacer (ITS1) of the nuclear ribosomal gene and its comparison with related species are described here. The nucleotide sequence of the ITS1 region flaking the 5.8S rRNA of this species and other related species are also given here.