Sequence and Phylogenetic Analysis of rDNA ITS of Three Eimeria Species in Rabbit

In order to study whether the internal transcribed spacers (ITS) sequence could be used as a molecular marker for the species identification of rabbit coccidian, the rDNA ITS of Eimeria intestinalis, Eimeria flavescens and Eimeria magna were amplified by polymerase chain reaction (PCR), and were cloned into pGEM-T Easy vector subsequently. The positive recombinant plasmids were identified by PCR and then sequenced. By sequence comparison and comparative analysis with the relative sequences of rabbit Eimeria spp. available in GenBank, the results showed that the lengths of Eimeria intestinalis, Eimeria flavescens and Eimeria magna were 1065, 1009 and 1047 bp, respectively, and the sequence homologies with the same species sequences were 99.2%, 99.0% and 94.5%, respectively, while were 55.3% to 82.1% compared with corresponding sequences of other different species sequences. The phylogenetic analysis using software Mega 5.0 showed that all rabbit coccidia clustered together in a clade, which was divided into two sister lineages, corresponding to the presence or absence of oocyst residuum. The result demonstrated ITS could be used as a molecular marker for the species identification of rabbit coccidia.     

河南商丘地区棉花黄萎病菌分离鉴定和致病力分析

为探讨河南商丘地区棉花黄萎病菌的致病型群体变异,对该地区棉花上分离的8株单孢菌株的菌落形态、显微结构、致病力、ITS序列、系统进化及菌体蛋白等方面进行了研究。结果表明:这些菌株均属于棉花黄萎病菌Verticilliumdahliae;系统进化树显示8株菌株并没有聚在同一进化枝上;8株黄萎菌菌株存在致病力差异,SQ4菌株致病力最强,属于落叶型,而其它致病力较弱的7个菌株属于非落叶型;不同致病力的菌株间蛋白谱带存在差异。     

Genetic characteristics of Streptococcus dysgalactiae isolated from cage cultured cobia,Rachycentron canadum (L.)

Disease outbreaks occurred during 2007–2013 in Taiwan with 2.5–10% mortality among the cage cultured cobia, Rachycentron canadum (L.), characterized by the presence of polyserositis, pericarditis and peritonitis. The micro‐organisms isolated from internal organs were Gram‐positive cocci. The isolates were confirmed as Streptococcus dysgalactiae by a polymerase chain reaction assay that yielded the expected specific 259 bp amplicon. Additionally, partial sequence of the 16S–23S rDNA intergenic spacer region of the GCS strain isolates from fish was also compared and produced 100% sequence identity with S. dysgalactiae (GenBank accession number AB252398 ). The genetic characterization was then determined by pulsed‐field gel electrophoresis (PFGE) analysis. Based on PFGE, the Apa I or Sma I digestion patterns of chromosomal DNA of these isolates were grouped into three main clusters. Taiwanese strains were divided into two clusters, and the tet(M) gene was detected in cluster 1 (pulsotypes: A1–A2 and S1–S3), but not in cluster 2 strains (pulsotypes: A3–A4 and S4–S5). Three Japanese strains from amberjack, Seriola dumerili (Risso), were grouped into cluster 3 (pulsotypes: A5–A7 and S6–S8) and displayed no mortality to cobia in the challenge experiment. Conversely, Taiwanese strains from cobia and snubnose pompano, Trachinotus blochii (L.), displayed a mortality rate of 50–87.5% in cobia.     

小耳花猪蛔虫ITS及5.8S rDNA序列扩增与分析

以分离自小耳花猪消化道内的蛔虫为研究对象,提取虫体的DNA片断,然后用引物对核糖体DNA(rDNA)的内转录间隔区ITS-1、ITS-2及5.8S序列进行PCR扩增。结果显示,目的片段ITS总长为1441 bp,2个不同样品之间的I TS序列没有差异。通过BLAST检索,与相关蛔虫I TS序列进行比较,发现与拜林蛔线虫(Bayl i sascari s t ransfuga)、猪蛔虫(Ascari s suum)和人蛔虫(Ascari s l umbri coi des)的ITS序列相似性分别为88%、98%和86%,与其他蛔虫的相似性均小于90%。     

西藏自治区首次发现螟黄赤眼蜂

为发掘西藏地区本土的赤眼蜂资源,本研究利用灭活米蛾卵卡在西藏林芝嘎玛地区的梨园采集到一个赤眼蜂种群。采用ITS2序列比对鉴定所采集的赤眼蜂种类,并测定该蜂的生物学指标。结果表明：西藏林芝嘎玛地区所诱集的赤眼蜂鉴定为螟黄赤眼蜂Trichogramma chilonis Ishii,羽化率为80%,雌性比为57.14%,耐饥饿时间为1.08 d,后足径节长度为157.6μm。本研究将为发掘西藏地区赤眼蜂资源提供参考,并为利用赤眼蜂在西藏自治区开展生物防治实践奠定基础。     

分子标记用于赤眼蜂分子监测的研究

通过对松毛虫赤眼蜂（Trichogramma dedrolimi Matsumura）以及玉米螟赤眼蜂（Trichogramma ostriniae Pang et Chen）rDNA-ITS2基因的克隆测序，并联机检索GenBank核酸序列库中其它赤眼蜂的相关序列，利用核酸分析软件找到松毛虫赤眼蜂和玉米螟赤眼蜂ITS2序列中一段有鉴别意义的标志，并据此设计出蜂种的特异PCR引物，实现了松毛虫赤眼蜂和玉米螟赤眼蜂rDNA特异区带的PCR扩增。研究表明该特异区带具有种的特异性。我们认为该分子标记技术可用于目前我国生防中常用的松毛虫赤眼蜂和玉米螟蒌眼蜂的分子监测，如田间种群动态监控和寄生效果评估。     

库姆塔格沙漠北界阿奇克谷地土壤可培养细菌多样性

为明确库姆塔格沙漠北界阿奇克谷地的土壤细菌多样性,探索该地区土壤微生物与植物群落的相互作用关系,采用NA、R2A培养基分离培养土壤细菌,通过16S rDNA序列系统发育分析鉴定菌株。结果表明,R2A平板菌落数量和种类较多;发现13株潜在新种菌株,已知菌种分属3大类群、4属、20种,厚壁菌门(Firmicutes)为优势类群,芽胞杆菌属(Bacillus)为优势属,枯草芽胞杆菌(Bacillus subtilis subsp.inaquosorum)为优势菌种。与其他类似环境相比,阿奇克谷地土壤中可培养细菌生物量偏小,可培养的细菌多为抗性、耐性强的极端微生物。该地区特有的微生物资源、功能微生物和新菌资源丰富,为防沙治沙、微生物菌剂开发等研究提供物质基础。本研究结果对库姆塔格沙漠的自然生态保护以及极端环境微生物资源的应用具有重要意义。     

基于ITS序列探讨西瓜种下分化

【研究目的】对源自中国和非洲的15个西瓜Citrullus lanatus的核糖体基因内转录间隔区(in-ternal transcribed spacers,ITS)全序列进行比较分析。供试材料包括Mucosospermus亚种、Vulgaris亚种、源自中国的3个品种、源自加纳的3个品种和1个来自布基纳法索的品种。【结果】西瓜种内总变异位点27个,占总碱基数的5.93%,6个信息位点,占总碱基数的1.32%。不同品种西瓜的ITS序列全长455～473bp,G+C含量61.49%～63.41%。【结论】从ITS序列全长构建的系统树可以看出西瓜具有地理分化现象,中国西瓜和非洲西瓜分属不同分支。此研究对西瓜的引种、育种以及研究西瓜的地理演化都具有一定的指导意义。     

广西北海红树林土壤放线菌的分离与鉴定

[目的]鉴定广西北海红树林土壤放线菌的物种多样性.[方法]从广西北海红树林土壤中分离、筛选10株典型放线菌菌株,提取基因组DNA,进行16S rDNA PCR扩增与测序,并构建进化树.[结果]通过Blast比对,10株放线菌属于2个属,其中8株为链霉菌属(80%),2株为拟诺卡氏菌属(20%).[结论]广西北海红树林土壤蕴减着种类丰富的放线菌. Abstract： [Objective] The aim was to identify the species diversity of Actinomycetes from Mangrove forest in Beihai,Guangxi Province.[Method]10 strains of typical Actinomycetes were isolated from Mangrove forest soil,and the Actinomycetes genomic DNA was successful extracted.16S rDNA was amplified by PCR and sequenced by Sanger dideoxy sequencing method.[Rcsult]All the sequences were blasted in genbank,eight strains belonged to the genus of Streptomyces (80%),and two strains belonged to the genus of Nocardiopsis (20%).[Coacluslon]There are many different Actinomycetes species in Mangrove forest soil samples in Beihai,Guangxi Province.     

RNA- and DNA-based profiling of soil fungal communities in a native Australian eucalypt forest and adjacent Pinus elliotti plantation

Total and active soil fungal communities in a native eucalypt forest and first rotation Pinus elliotti plantation were investigated by direct extraction of DNA and RNA from soil. Terminal restriction fragment length polymorphism (T-RFLP) analysis of internal transcribed spacer (ITS) and 18S rRNA profiles indicated that total and active fungal communities differed significantly in both forest types. This was supported by DGGE profile analysis on an individual plot basis for both forest types and when groups in the canonical analysis were redefined to allow comparison between forest types. Analyses of both ITS and 18S T-RFLP profiles indicated that conversion from native eucalypt forest to P. elliottii plantation may significantly alter total and active soil fungal communities. ITS DGGE (DNA) and 18S (RNA) profiles also suggested differences in fungal communities in the two forest types. No significant separation of the fungal communities in the two forest types was observed, however, when ITS DGGE (RNA) profiles were compared. Overall, the data suggest that conversion from native eucalypt forest to P. elliottii plantation at the Beerburrum State Forest in subtropical Australia has significantly altered soil fungal communities.