利用28S rDNA D1/D2区和ITS rDNA序列鉴定甜瓜白粉病病原菌

为了明确宁夏干旱带压砂甜瓜白粉病病原菌,从病原菌分生孢子中提取DNA,PCR扩增ITSrDNA和28S rDNA D1/D2区段,测序后进行BLAST比对.结果表明,病原菌的ITS rDNA和28SrDNA D1/D2序列与菜豆叉丝单囊壳白粉菌Podosphaera phaseoli、凤仙花又丝单囊壳白粉菌P.bal-saminae、菊科叉丝单囊壳白粉菌P.fwca、苍耳单囊壳白粉菌P.xanthii、瓜类单囊壳白粉菌P.fuligi-nea等叉丝单囊壳属Podosphaera的多个种的ITS rDNA和28S rDNA D1/D2序列之间相似度均大于99%,鉴定甜瓜白粉病病原菌为叉丝单囊壳属Podosphaera.     

Grouping of Colletotrichum Species in Japan Based on rDNA Sequences

Internal transcribed spacers (ITS) of the ribosomal RNA gene (rDNA) were sequenced for 236 isolates covering 25 Colletotrichum species collected in Japan. The Japanese isolates could be grouped into 20 ribosomal groups (RGs) based on the sequences of ITS1, correlating the species identified by the morphology. Colletotrichum gloeosporioides sensu lato separated into three RGs that were morphologically different. Colletotrichum destructivum, C. linicola and C. higginsianum were possibly conspecific. Colletotrichum dematium sensu lato including C. capsici and other species that produce falcate conidia except for graminicolous ones were separated into three RGs that were difficult to distinguish morphologically. In the phylogenetic study using ITS2 and the 28S rDNA domain 2 region, topologies compiled by neighbor-joining and maximum-parsimony methods were almost the same, reflecting the conidial morphology. The phylogenetic group 1 (PG1) produced conidia with acute ends, e.g., C. acutatum, C. destructivum and C. graminicola; PG2 produced those with obtuse ends, e.g., C. gloeosporioides, and C. orbiculare. Colletotrichum theae-sinensis, which produced the smallest conidia, was grouped as PG3, far from other species, indicating it should not belong to Colletotrichum. Grouping and phylogenetic analysis using ribosomal DNA was an effective tool to classify and identify Colletotrichum species without using morphology. Received 15 July 2002/ Accepted in revised form 12 November 2002     

甘肃小麦禾谷孢囊线虫rDNA-ITS和28S rDNA-D2/D3区序列特征及ITS-RFLP分析

禾谷孢囊线虫是危害禾谷类作物的重要病原,严重威胁我国小麦主产区的小麦产量和品质。利用通用引物对甘肃、河南、安徽禾谷孢囊线虫群体28SrDNA-D2/D3区和rDNA-ITS区进行PCR扩增和序列测定,利用UPG-MA方法分析了甘肃省7个种群、河南1个种群、安徽1个种群的禾谷孢囊线虫群体D2/D3区和ITS区的系统发育关系;用9种限制性内切酶对7个甘肃禾谷孢囊线虫群体的rDNA-ITS区进行了RFLP分析。结果表明:中国甘肃的禾谷孢囊线虫rDNA-D2/D3区片段长度约为780bp,rDNA-ITS片段长度约为1040bp。7个甘肃禾谷孢囊线虫群体、1个河南安阳群体、1个安徽蚌埠群体的D2/D3区和新西兰的H.aucklandica群体亲缘关系很近;其ITS区同澳大利亚的H.australis、北京通州的H.avenae(AY148382)的亲缘关系很接近。RFLP分析表明,9种限制性内切酶酶切禾谷孢囊线虫群体的rDNA-ITS共产生了22个酶切片段,不同酶切的RFLP分布型在7个种群间没有差异。甘肃省禾谷孢囊线虫群体的rDNA-ITS区具有高度的保守性,与中国的C型群体相近,但不同于欧洲的A型群体和印度的B型群体。这是首次报道甘肃CCN种群分子特征。     

马铃薯腐烂茎线虫28S rDNA-D2/D3区序列分析

 我国危害甘薯的马铃薯腐烂茎线虫rDNA-ITS基因片段长度存在A型(900bp)和B型(1100bp)2个类型,A型腐烂茎线虫群体的ITS片段比B型群体少200bp。为了进一步研究这2种类型群体的发育关系,本文用线虫通用引物D2A和D3B对来自国内的21个马铃薯腐烂茎线虫群体和1个韩国的马铃薯腐烂茎线虫28SrDNA-D2/D3区进行了PCR扩增,均产生大小一致的片段,长度约为780bp,克隆、测序后用DNAMAN5.2软件和MEGA4软件进行分析比对,结果表明我国21个马铃薯腐烂茎线虫群体28SrDNA-D2/D3区只有20余个碱基的差异,相似率达97.2%。基于UPGMA构建的系统发育树很好地区分了马铃薯腐烂茎线虫A型和B型群体,展现出了群体的来源及发育关系。     

Comparative Analysis of Japanese and Foreign Strains of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> Based on 16S Ribosomal RNA Gene Sequences

We determined nearly the complete sequences of the 16S ribosomal RNA gene (rDNA) for Japanese strains of R. solanacearum. The comparison of 1471 nucleotide positions separated the Japanese strains into two groups, group 1 with biovars 1, 2, 3 and 4 strains which belonged to race 1, and group 2 with biovar 2 strains corresponding to race 3. Group 1 strains all had identical sequences, and strains representing the four biovars within the group did not differ from each other. Group 2 strains had characteristic nucleotides which differed at seven positions from group 1 strains. Comparative analysis of Japanese and foreign strains based on 16S rDNA sequences showed that Japanese group 1 was closely related to Asian and Australian biovars 3, 4 and 5, and belonged to the known division 1. Japanese group 2 was homogeneous to Indonesian biovars 2 and N2 in subdivision 2b. Since the differences in the nucleotides corresponded to restriction sites for the AluI, RFLP analysis of PCR-amplified 16S rDNA efficiently differentiated not only Japanese group 1 from group 2, but also differentiated three types of foreign strains which differed in biovar and geographic origin. Received 26 July 1999/ Accepted in revised form 19 November 1999     

Phylogenetic relationships between the lettuce root rot pathogen Fusarium oxysporum f. sp. lactucae races 1, 2, and 3 based on the sequence of the intergenic spacer region of its ribosomal DNA

The genetic relationship between the vegetative compatibility groups (VCGs) and between physiological races of Fusarium oxysporum f. sp. lactucae (FOL), the causal pathogen of lettuce root rot, was determined by analyzing the intergenic spacer (IGS) region of its ribosomal DNA. A total of 29 isolates containing a type strain were tested: 24 Japanese isolates, 2 Californian isolates, and 3 Italian isolates. Three races (races 1, 2, and 3) were found in Japan, and race 1 was also distributed in California and Italy. Races 1, 2, and 3 each belonged to a distinct VCG: VCG-1, VCG-2, and VCG-3 (VCG-3-1, VCG-3-3), respectively. Phylogenetic (neighbor-joining) analysis of the IGS sequences revealed that races 1, 2, and 3 coincided with three phylogenetic groups (PG): PG-1, PG-2, and PG-3, respectively. These results indicate that the three races are genetically quite different and have a strong correlation with VCGs and phylogenetic groupings. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession no. AB195218