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1.
本文分析了梅奇酵母属Metschnikowia chrysoperlae JA30与其同属已知菌种的属内种间系统发育关系。提取M.chrysoperlae JA30的基因DNA,对其内转录间隔区(ITS)序列进行PCR扩增和序列测定。采用BLAST方法将内转录间隔区(ITS)序列在GenBank中进行同源搜索,运用邻接距离距阵法,建立梅奇酵母属共33种的ITS序列的系统发育树,并探讨了其亲缘关系。结果表明,梅奇酵母属的M.chrysoperlae、M.pulcherrima、M.fructicola在ITS基因序列上的差异表现为种内变异水平,这与传统分类的结论存在分歧。  相似文献   

2.
正在众多的花卉品种中,有一些花卉的形态十分相似,如不仔细分辨,一般人很难区分。现将几种常见而又相似的花卉如何区分,作一些简要介绍。矮牵牛与牵牛矮牵牛和牵牛二者名虽相近,花形、果形也相同,但它们并无亲缘关系。二者的差别相当明显,即矮牵牛为茄科矮牵牛属植物,多年生草木,株高仅为40至60厘米,花期为4月到霜降,是布置花坛和盆栽的好料;而牵牛属旋花科牵牛属,为1年生缠绕茎草本,茎攀援而生,可长至3米左右,花期6至10月,适于做花架栽培,用于屋顶及廊架的绿化。  相似文献   

3.
在青篱竹属Arundinaria 系统分类中由于对该属的形态分类标准取舍不一,至今仍存在着严重的意见分歧。本文采用PCR 扩增产物直接测序的方法分析青篱竹属Arundinaria 及近缘属(如苦竹属Pleioblastus、矢竹属Pseudosasa、少穗竹属Oligostachyum、巴山木竹属Bashania、肿节竹属Clavinodum 等)有关争议属的代表种或模式种(毛竹为外类群)等18 种竹种的核糖体DNA ITS 区段序列和叶绿体DNA trnL-F 序列。分别对ITS 序列、trnL-F 序列和ITS 与trnL-F 的组合序列进行系统发育分析。结果表明,和trnL-F 区段序列相比,核糖体DNA ITS 区段序列有较高的系统发育信息位点。通过最简约分析产生的ITS 树、trnL-F 树和ITS 与trnL-F 的组合序列树表明,所得树形基本相似。供试竹种(斑苦竹A. oleosa、仙居苦竹A. hsienchuensis、茶秆竹A. amabilis、长叶苦竹A. chino、苦竹A. amara、宜兴苦竹A. yixingensis、菲白竹A. fortunei、翠竹A. pygmaea、大明竹A. gramineus、巴山木竹A. fargesii、冷箭竹A. faberi、凤竹A. hupehense、鼓节矢竹P. japonica cv. Tsutsumiana、矢竹P. japonica、短穗竹B. densiflorum、肿节竹A. oedogonata、少穗竹A. sulcata)形成一个单系类群,且分为2 个不同的分支。图3 表2 参11。  相似文献   

4.
以开红花和黄花木棉属木棉Bombax malabaricum的14株个体的核糖体DNA ITS序列测序结果,构建了系统发育树,并选取与木棉属较近亲缘关系的吉贝属Ceiba Mill.emend.Gaertn.、轻木属Ochroma Swartz的种作为外类群,基于Maximum likelihood analysis(最大似然法)、Maximum parsimony analysis(最大简约法)和Bayesian inference(贝叶斯方法 )对木棉属红花和黄花的木棉个体植株进行分子系统发育重建。结果表明,开红花木棉和开黄花木棉的植物学特征描述基本一致,来源于不同地点或同一地点供试木棉植株分子序列与Genbank中记录的木棉的ITS全长序列相似度为99%,与同属的Bombax buonopozense亲缘关系较近,并且与ITS序列对红花木棉和黄花木棉所有物种以100%的Bootstrap聚在一起。对两者的主要植物学性状和分子生物学基因序列综合评价,确认红花木棉和黄花木棉同属一个种。  相似文献   

5.
柳属植物ITS序列分析   总被引:2,自引:0,他引:2  
本文分析了垂柳、曲枝垂柳、旱柳和龙爪柳以及其他柳属植物属内种间系统发育关系。提取垂柳、曲枝垂柳和龙爪柳基因组DNA,对其内转录间隔区(ITS)序列进行PCR扩增和序列测定。采用BLAST方法将植物内转录间隔区(ITS)序列在GenBank中进行同源搜索,应用邻接距离距阵法,建立柳属共19种的ITS序列的系统发育树,并探讨了其亲缘关系。结果表明,垂柳、曲枝垂柳、旱柳和龙爪柳应属于同一种,其关系为种内变异水平,与经典分类方法中垂柳与旱柳为两个不同种存在分歧。  相似文献   

6.
通过克隆和测定核糖体DNA内转录间隔区(Ribosomal DNA internal transcribed spacer,rDNA,ITS)基因序列,研究红榉不同种源间的亲缘关系。测定结果表明:红榉5个种源的ITS序列、ITS1和ITS2长度变异范围分别为561~623 bp、157~210 bp和215~222 bp。软件DNAMAN、遗传距离和UPGMA法系统发育树分析结果表明:南京种源与赣州种源的亲缘关系较近,湖州种源与怀化种源的亲缘关系较近;滁州种源单独聚为一类,与湖州、怀化种源的亲缘关系较远。  相似文献   

7.
提取刚竹属中毛金竹、毛竹、龟甲竹、人面竹、紫竹、白哺鸡竹、黄槽竹、芽竹、蓉城竹、红壳雷竹、假毛竹、金竹、黄纹竹等13个竹种的DNA,对其内转录间隔区(ITS)序列以及maturase K(mat K)序列进行PCR扩增和序列测定。构建了与其亲缘关系接近的刚竹属6个种、牡竹属3个种共22个竹种基于ITS序列的系统发育树和刚竹属7个种、牡竹属3个种共23个竹种基于mat K的系统发育树,并探讨了其亲缘关系。结果显示,实验所用的大部分竹种分类结果与传统分类类似,但黄纹竹、毛竹与其他刚竹属竹种的遗传距离较远,却与牡竹属的竹种聚类,与传统分类相左。通过测序分析认为其为杂种的可能性较高,并由此推测杂种的存在是造成竹子分子分类困难的一大原因。  相似文献   

8.
利用外部形态和解剖结构特征进行对比,选出樟子松Pinus sylvestris var.mongolica 2个类似块菌属Tuber的外生菌根类型,再采用CTAB法对其进行基因组DNA提取,并利用真菌特有引物ITS1F和ITS4,在PCR基础上对其r DNA的ITS区段进行碱基序列测定,并与Gen Bank数据库中已知的相关序列进行Blast比对得出:类型1,类型2均为块菌属的外生菌根,类型1可鉴定到种,为辽东块菌Tuber liaotongense。  相似文献   

9.
文章对泰国清迈蘑菇属真菌进行rDNA ITS区段序列测定和序列特征比较分析,并基于ITS序列进行核酸数据库GenBank同源性检索比对,最终构建系统发育树,旨在为从分子水平上探讨蘑菇属真菌的系统发育、亲缘关系和遗传多样性提供参考。  相似文献   

10.
记述采自中国境内危害白榆的突瓣叶蜂属(膜翅目:叶蜂科)一新种:白榆突瓣叶蜂 Nematus pumila Liu,Li & Wei,sp.nov.。该种幼虫取食白榆的叶片,当虫口密度较大时,可将寄主叶片蚕食殆尽,对寄主产生严重危害。本研究对林业有害生物的鉴定、监测与防治工作奠定了基础。新种与申氏突瓣叶蜂 N.sheni Wei,1999十分相似,二者的主要区别是:新种上唇、前胸背板和翅基片完全黑色;前足胫节、中足胫节基部3/4和后足胫节基部3/5白色;单眼后区宽长比为1.9;前翅无Rs脉;锯腹片15锯刃,锯刃内侧亚基齿明显,外侧亚基齿粗大;1-11节缝具刺毛带,刺毛带最宽处约2/3于锯节宽;锯根0.7倍于锯端;申氏突瓣叶蜂上唇黄白色;前胸背板后缘、翅基片基小部黄褐色;前中足胫节腹侧大部和背侧基部1/4、后足胫节基部1/3黄白色;单眼后区宽长比约等于3;前翅Rs脉痕状;锯腹片23锯刃,内齿亚基齿不明显,外侧亚基齿细小;2-15节缝具刺毛带,刺毛带最宽处约1/3于锯节宽;锯根0.4倍于锯端。采用DNA试剂盒法提取了新种基因组DNA,测序获得COⅠ基因序列,长度为810 bp。另外,从GenBank数据库中下载突瓣叶蜂属、槌缘叶蜂属、厚爪叶蜂属已知种类COⅠ序列10条,与新种序列构成COⅠ序列数据集,分析其碱基组成,结果显示: A+T含量均大于G+C含量,且均高于65%;基于贝叶斯法构建了COⅠ基因系统发育树,树图结果显示:白榆突瓣叶蜂与突瓣叶蜂属内其他6个已知种聚为一支,构成单系群。分子分析结果与形态鉴定结果一致,均支持新种成立。新种模式标本保存于湖南长沙中南林业科技大学昆虫模式标本室。  相似文献   

11.
基于ITS和cpDNA序列的梭梭和白梭梭物种分化   总被引:2,自引:0,他引:2  
【目的】基于nrDNA和cpDNA 2种基因,探讨中国同域分布的梭梭和白梭梭的物种分化,分析2种基因的单片段及其片段组合对2种植物的物种鉴别力,并分析生态位分化的程度及其对物种进化的影响,为梭梭和白梭梭的系统发育和谱系地理等研究提供基础数据。【方法】基于2个nrDNA序列(ITS2、ITS1-ITS4)和3个cpDNA非编码区序列(trnS-trnG、rps4和trnV),对古尔班通古特沙漠南缘同域分布的梭梭和白梭梭共30个个体进行序列分析;利用距离法(基于Kimura 2-parameter)研究2种植物的遗传分化;基于贝叶斯方法分析个体间的分子系统关系;并利用距离法、系统发育树法、BLAST比对法和诊断特征法评价ITS和cpDNA的单序列及其组合对2种植物的物种鉴别力。在此基础上,利用生态位分析软件ENMTools V1.4定量分析梭梭与白梭梭生态位分化的程度。【结果】1)对于梭梭和白梭梭,2个ITS序列拼接比对的总长均为1 117 bp,G+C含量平均分别为34.74%和34.82%,总的信息位点数分别占片段总长的2.33%和1.43%; 3个cpDNA序列拼接比对的总长均为2 344 bp,G+C含量平均分别为59.62%和59.39%,信息位点数分别占片段总长的0.68%和0.43%。2)基于ITS和cpDNA序列组合构建的贝叶斯系统发育树都表明梭梭和白梭梭各聚为一支。3)基于ITS和cpDNA序列组合计算的2种植物的种间最小遗传距离均大于种内的最大遗传距离,并且种间种内都存在1个明显的距离间隙,表明本研究所用的ITS和cpDNA的序列组合可以作为鉴定梭梭和白梭梭的DNA条形码序列。4)基于4种方法评价的ITS和cpDNA序列对梭梭和白梭梭的物种鉴别力表明:2个ITS的单序列及其序列组合的鉴别率均为100%; cpDNA序列中,rps4的单序列及其与trnS-trnG,trnV的两两序列组合的鉴别率均为0,而trnS-trnG,trnV的单序列及其序列组合,以及trnS-trnG+trnV+rps4组合的鉴别率均为100%。5)生态位参数D值和I值的观察值和一致性检验的模拟值都集中在0.65和0.90左右,表明梭梭和白梭梭的生态位分化不明显。【结论】基于ITS和cpDNA序列组合,梭梭和白梭梭物种间的遗传分化明显,分子系统关系清楚; ITS和cpDNA序列组合都可以作为鉴定梭梭和白梭梭的DNA条形码序列;梭梭和白梭梭的生态位分化不明显,说明生态因素很可能对2种植物的分化与进化没有起到明显的作用,2种植物很可能也具有相近的进化历史。本研究的结论可以为梭梭和白梭梭的系统发育、遗传和进化研究提供重要的理论依据和数据支撑。  相似文献   

12.
Pseudodidymella fagi is a leaf blotch pathogen of Fagus crenata in Japan. This pathogen is now reported for the first time on F. sylvatica in Switzerland and Germany. Species identity was verified by morphological assessment of the asexual and sexual morphs and by comparing the internal transcribed spacer (ITS) sequence of the type material from Japan and of freshly collected samples from Europe. ITS sequences proofed to be completely identical. The asexual morph Pycnopleiospora of Ps. fagi is formed on necrotic leaf spots during summer and autumn. In early spring, its sexual morph is formed in the litter of F. sylvatica. The connection between sexual and asexual morphs was verified by sequencing the ITS region of single conidium and ascospore isolates. The pathogenicity of Ps. fagi on F. sylvatica was tested by inoculations on detached leaves in vitro and Koch's postulates were fulfilled. The second pathogen we report in central Europe for the first time is Petrakia deviata, which causes a leaf blotch disease of field maple (Acer campestre). This species was collected only once in the central Caucasus region in 1929 and was never found again after its first discovery. Now it was rediscovered in two different locations in Switzerland on field maple (A. campestre) and on the new host Norway maple (A. platanoides). Species identity was verified by morphology and by comparison with the ITS sequence of the holotype specimen and freshly collected samples. Phylogenetic analysis based on ITS sequences suggested that Ps. fagi and P. deviata are closely related to each other. Whether these species were simply overlooked so far, profit from climate change, or represent newly introduced invasive species remains to be studied. Moreover, deeper phylogenetic analysis using multiple sequence markers should be conducted to verify species identities.  相似文献   

13.
A method for identification and typing of wood rotting fungi using the melting temperature [T(m)] of DNA fragments coding rRNA (rDNA) was examined. The T(m)s of four DNA fragments, inter transcribed spacer (ITS) I, ITS II, and two partial fragments of 28S rDNA from each of 20 species of wood rotting fungi, were measured by melting curve analysis. The T(m) variation was large enough between species to enable identification based on the T(m) values. A pair of T(m)s of the ITS I region (between the primers ITS1 and ITS2) and the ITS II region (between the primers ITS3 and ITS4) had the highest resolution for identifying wood rotting fungi. To assess about the diversity of the T(m), intraspecific diversity of these DNA fragment sequences was evaluated using test strain sequences and data from the GenBank/EMBL/DDBJ biological database. The intraspecific diversity of T(m) was considered to be small because the nucleotide diversity of each fragment was small within the species.  相似文献   

14.
Poplar rusts due to Melampsora larici‐populina (Mlp), M. allii‐populina (Map) and M. medusae f. sp. deltoidae (Mmd) are the most serious disease in Europe on cultivated poplars, that is, Populus × euramericana and P. × interamericana hybrids. These pathogenic species can be identified by the observation of morphological characteristics of urediniospores but this method is not appropriate for high‐throughput analysis and cannot be used on other spore stages, such as aeciospores or teliospores, that are morphologically similar. The aim of this study was to develop a rapid and sensitive molecular method based on PCR amplification that was able to specifically detect these species on various hosts for routine analysis. Three primer pairs ITS‐MLP‐F/ITS‐MLP‐R, ITS‐MAP‐F/ITS‐MAP‐R and ITS‐MMD‐F/ITS‐MMD‐R were designed within the internal transcribed spacer (ITS) sequences of ribosomal DNA to target Mlp, Map and Mmd, respectively, and their specificity were confirmed on a wide range of isolates and species. ITS‐MLP‐F/ITS‐MLP‐R and ITS‐MAP‐F/ITS‐MAP‐R primers proved to be highly specific to Mlp and Map, respectively, whereas ITS‐MMD‐F/ITS‐MMD‐R cross‐reacted with DNA from M. larici‐tremulae and M. pinitorqua. However, these species are not pathogenic on cultivated poplars that all belong to sections Aigeiros and Tacamahaca of the genus Populus. Specific Mmd primers proved to be very sensitive as a positive signal could be obtained with DNA extracts from 6 target urediniospores mixed with 800 000 urediniospores of Mlp. An internal amplification control (IAC) was included to discriminate false negative results due to the potential presence of inhibitory compounds in DNA extracts. ITS‐MMD‐F/ITS‐MMD‐R primers are therefore efficient for the detection of the quarantine pathogen Mmd on samples collected on poplar or larch and are fit for use in official tests. This new PCR assay has been used in routine for ten years, and Mmd has hitherto never been detected in commercial poplar nurseries in France.  相似文献   

15.
  • ? Powdery mildew is a major fungal disease of oaks in Europe. Recent studies using internal transcribed spacer (ITS) sequences suggested the presence of four different lineages (putative species). The objective of the study was to investigate the spatial distribution of these lineages/species and, in particular, to test the hypothesis of a spatial differentiation, at various scales: regional (France), altitudinal (a Pyrenean transect) and local (within a forest plot).
  • ? Detection methods for the four ITS types were developed: (1) single strand conformation polymorphism analysis (SSCP); (2) PCR amplifications, for which specific primers were designed. SSCP proved to be efficient for the detection of Erysiphe alphitoides and E. quercicola types. In contrast, the rarer ITS types corresponding to E. hypophylla and Phyllactinia guttata (sensu lato) were only detected by specific amplification.
  • ? The study confirmed the strong predominance of the ITS sequence associated with E. alphitoides at all spatial scales (with a frequency higher than 80%). Isolates presumably belonging to E. quercicola (i.e. with same ITS type), a recently described species not yet recorded in Europe, were also found in all French regions at a significant frequency (15% at national level).
  • ? No pattern of spatial differentiation between the putative species could be demonstrated: E. alphitoides was often found in association with different ITS types in the same region, the same tree, and even in the same lesion.
  •   相似文献   

    16.
    Bursaphelenchus fungivorus is reported for the first time in Portugal, identified as associated with Pinus pinaster bark and characterized on the basis of morphological and morphometrical characters for this species. Species identification was confirmed using restriction fragment length polymorphism analysis and sequencing of the internal transcribed spacer (ITS) regions of ribosomal DNA. Intraisolate genetic variability was detected among ITS sequences of the Portuguese B. fungivorus isolate. Phylogenetic analysis, obtained from multiple sequence alignment between ITS sequences of Bursaphelenchus species, revealed that the Portuguese B. fungivorus isolate clusters with other B. fungivorus isolates, forming a separate group close to B. seani highlighting a molecular proximity of these two species.  相似文献   

    17.
    采集巨桉林下马勃子实体,培养其菌丝体,提取子实体及菌丝体基因组DNA,进行rDNA-ITS区序列的扩增、克隆和序列测定,并对rDNA-ITS不同区域作序列分析,首次构建马勃的系统发育树.结果表明:野外采集获得马勃子实体l0种,其中成功培养6种,测序结果表明马勃rDNA-ITS区长度在607~766 bp之间,系统发育分析表明硬皮马勃属(Scleroderma)与豆包菌(Pisolithus)亲缘关系较近,秃马勃属(Calvatia)、马勃属(Lycoperdon)及横膜马勃属(Vascellum)之间亲缘关系较近,ITSl-5.8S rDNA-ITS2区可建立马勃类真菌属间系统发育树,ITS2区可用于建立马勃类真菌属内系统发育树,3个待定种硬皮马勃Scleroderma sp.11-1,Scleroderma sp.2-2和Scleroderma sp.5-2为金黄硬皮马勃(S.aurantium)的可能性较大.此研究可为探讨巨桉人工林下外生菌根种类与作用机制、马勃分类系统学及菌丝体的开发研究奠定基础.  相似文献   

    18.
    用形态分类和ITS序列分析法对湖南桂东黄菌进行了鉴定,用基于Bayians理论和Markov chain Monte Carl(MCMC)算法的MrBayes3.1.2软件,将桂东黄菌与Genbank中牛肝菌科40个物种85个ITS序列进行系统发育树分析.结果表明,以欧洲缘盖牛肝菌(Boletus appendicul...  相似文献   

    19.
    Lophodermium piceae, the dominant needle endophyte of Norway spruce (Picea abies), is known to be extremely diverse. This work aimed to test the possible occurrence of cryptic species within the morphological species L. piceae. Genetic variation in 36 South Finnish L. piceae isolates originating from six localities was investigated by comparing DNA sequences of three genetic markers. One of the markers was the internal transcribed sequence (ITS) of the ribosomal DNA and the other two (LP1 and LP2) were based on sequence characterized amplified regions designed for L. piceae. The LP2 marker could be detected only from isolates of L. piceae but not from 20 other ascomycete species tested. This sequence, therefore, is considered as a species‐specific marker for L. piceae. For comparison, ITS sequences of isolates representing two other Lophodermium species, L. pinastri and L. seditiosum, were also investigated. In a neighbor‐joining analysis of ITS sequences all L. piceae isolates fell into one cluster, which was clearly separate from those of L. pinastri and L. seditiosum. Dendrograms of the three markers were incongruent indicating that the L. piceae population examined consisted of a single phylogenetic species. No geographical differentiation was observed. Our results confirm that L. piceae is a genetically highly diverse endophytic species.  相似文献   

    20.
    杓兰属植物具有很高的观赏和药用价值,但因被过度采集,已成为濒危植物。菌根真菌是杓兰栽培与繁育是否成功的关键因子。本实验采用rDNA ITS序列扩增技术和MEGA软件对滇西北的云南杓兰和紫点杓兰的菌根真菌进行研究,结果表明:云南杓兰和紫点杓兰的菌根真菌存在显著的物种层面的专一性关系,即菌根真菌与杓兰属植物有较强的共生趋势。分离得到的菌根真菌序列已上传至 NCBI,登录号为 lcl51879、 lcl25153、lcl48033、 lcl38377、 lcl14203、 lcl24557、 lcl52287以及lcl46937。  相似文献   

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