冬虫夏草培养子实体ITS,5.8S的分析及系统发育研究

对野生冬虫夏草的人工培养子实体的ITS1,ITS2和5.8S间区序列进行了克隆,并结合已有的序列对ITS1,ITS2和5.8S进行了系统发育分析。结果表明,人工培养子实体与中华被毛孢(Hirsutella sinensis L95DBM-1)具有96%的同源性,在NJ邻接树上形成明显的一支,与虫草属其它类群种间具有明显差异,表明分离培养的虫草子实体为中华被毛孢。遗传距离分析结果显示,培养虫草序列与已知虫草在种内也存在一定差异,可能暗示虫草种群在不同区域具有一定的遗传分化。     

中国木蓝属两个分布新记录

报道并描述了采自云南南部西双版纳具有1小叶的新分布种美脉木蓝Indigofera caloneura Kurz和采自广东具有单叶的新分布种心叶木蓝I. cordifolia Heyne ex Roth.。在中国还有另外3个具有单叶的种类, 即单叶木蓝I. linifolia、刺荚木蓝I. nummularifolia和远志木蓝I. squalida。通过从植物习性、茎、叶的形态与大小及毛被、托叶、花序、花冠形状与毛被、花药、子房毛被、果实的形态与大小、所含种子数目等特征的显微观察, 比较了国产单叶类木蓝属Indigofera植物的形态学特征差异, 并针对茎、叶、毛被、花冠旗瓣和果实形状等5个最重要的特征进行了讨论。     

应用5．8SrDNA及ITS区序列分析链格孢种级分类

对9个链格孢小孢子种和3个大孢子种共20个链格孢菌株的5．8SrDNA及其两侧的ITS1区和ITS2区进行了序列分析。聚类分析结果表明形态差异大的种可以明确加以区分,而供试的9个小孢子种之间差异很小,不能根据对所选区段的序列分析加以区分,传统分类上的滨菊链格孢不属于Alternaria,其分类地位需进一步研究。     

基于形态和分子证据的大花忍冬复合群的种间关系(英文)

大花忍冬复合群主要分布在中国南部及周边地区,是忍冬属的主要疑难类群之一,包含大花忍冬、灰毡毛忍冬、细毡毛忍冬、菰腺忍冬和锈毛忍冬5种。该复合群是中国南部金银花的重要野生替代资源。本研究通过形态和生境观察、叶表皮微形态扫描和分子系统学分析相结合的方法,探讨了大花忍冬复合群的种间关系。结果表明,分子系统学方法没有解决该复合群之间的关系,仅灰毡毛忍冬的区分较为明显,基于形态学证据大致可将大花忍冬复合群分为以下3小群:菰腺忍冬以其独有的蘑菇状腺体与其他4种显著不同,毛被长度也明显长于其他种类;锈毛忍冬与大花忍冬的毛被较稀疏,但锈毛忍冬的花冠筒较短,气孔特征独特;灰毡毛忍冬与细毡毛忍冬的叶背密覆柔毛,二者的上表皮角质膜结构有差异。该复合群的物种形成和传播机制仍待深入研究。     

七种木蓝属植物的染色体研究

本文采用根尖压片法对豆科（Leguminosae）木蓝属（IndigoferaL.）植物的7个种：多花木蓝（Indigofera amblyantha Craib）、河北木蓝（I.bungeana Walp.）、滇木蓝（I.delavayi Franch.）、腺毛木蓝（I.scabrida Dunn）、四川木蓝（I.szechuensis Craib）、刺序木蓝（I.silvestrii Pamp.）、尖叶木蓝（I.zollingeriana Miq.）的染色体数目和核型进行了研究。结果表明：除了尖叶木蓝染色体数目为2n=4x=32（四倍体）外,其余6种木蓝的染色体数目均为2n=2x=16（二倍体）。尖叶木蓝和刺序木蓝核型分类为2A型,其余5种木蓝的核型均为1A型。种间核型差异很小。供试种主要包含中部着丝点区染色体。滇木蓝、腺毛木蓝、刺序木蓝、四川木蓝和尖叶木蓝的核型分析为首次报道。     

基于形态特征和ITS序列对7个鹅膏菌属菌株的分类鉴定

以采自浙江省丽水地区的7个鹅膏菌属菌株作为研究材料,在基于形态特征进行初步鉴定的基础上,对7种鹅膏菌的rDNAITS区段进行克隆测序和序列特征比较分析。进一步对ITS序列进行核酸序列数据库GenBank同源性检索比对,将从GenBank检索获得的9个最相似物种的ITS序列连同7种鹅膏菌的ITS序列一起作系统发育分析。结果表明:基于ITS序列对f6、f9和f493个菌株的分子鉴定支持了基于形态特征的鉴定结果,对f5的分子鉴定不支持形态鉴定的结果,f8为鹅膏菌属内某种,f66为鹅膏菌属内某种,并与Amanitafulva,A.atrofusca,A.orientifulva3种鹅膏菌的亲缘关系较近,f7与另外6种鹅膏菌的亲缘关系相差甚远。研究结果提示基于分子水平上的ITS序列分析不能单方面作为大型真菌分类鉴定的可靠依据,可以作为基于传统形态学分类鉴定的辅助参考依据。     

麻花艽和管花秦艽( 龙胆科) 之间自然杂交类型的分子验证

野外考察发现麻花艽( Gentiana straminea Maxim. ) 和管花秦艽( G. siphonantha Maxim . ex Kusn. ) 同域分布时存在大量形态位于二者之间的个体。经形态变异研究后发现它们可能是这两个物种之间的杂交后代。对两个亲本种以及假设杂交群体共55 个个体的核糖体ITS 序列和叶绿体trnS-G 序列的分析结果表明:中间形态个体是麻花艽和管花秦艽的自然杂交后代。此外分析了两个亲本种以及杂交群内个体间trnS-G和ITS 序列的变异状况以及分子标记结果与形态鉴定不一致的可能原因; 指出可能是杂交诱导的叶绿体基因组重组以及早期物种分化中的谱系筛选不彻底等原因造成了亲本种群体内序列变异的多样化。     

麻花艽和管花秦艽(龙胆科)之间自然杂交类型的分子验证

野外考察发现麻花艽(Gentiana stramineaMaxim.)和管花秦艽(G.siphonantha Maxim.ex Kusn.)同域分布时存在大量形态位于二者之间的个体。经形态变异研究后发现它们可能是这两个物种之间的杂交后代。对两个亲本种以及假设杂交群体共55个个体的核糖体ITS序列和叶绿体trnS-G序列的分析结果表明:中间形态个体是麻花艽和管花秦艽的自然杂交后代。此外分析了两个亲本种以及杂交群内个体间trnS-G和ITS序列的变异状况以及分子标记结果与形态鉴定不一致的可能原因;指出可能是杂交诱导的叶绿体基因组重组以及早期物种分化中的谱系筛选不彻底等原因造成了亲本种群体内序列变异的多样化。     

基于cpDNA和ITS序列对庭藤复合群的居群遗传学研究

采用2个叶绿体基因组片段(ndh J-trn F和trn D-trn T)和核基因组ITS序列对分布于中国东部和相邻地区的庭藤复合群(Indigofera decora complex)的24个居群进行分析,探讨该复合群的遗传多样性、单倍型分布、遗传结构和居群历史动态。cp DNA和ITS序列分析表明,庭藤复合群的单倍型多样性较高,而总体遗传多样性偏低(cp DNA片段:Hd=0.778,π=0.00123;ITS:Hd=0.909,π=0.01290);皖浙地区居群的遗传多样性相对较高,推测其可能是该复合群遗传多样性的分布中心。遗传变异主要存在于复合群居群间,种内居群间遗传分化较大(cp DNA片段:FST=0.848;ITS:FST=0.787);基因流较小(cp DNA片段:Nm=0.228;ITS:Nm=0.241),主要是地理隔离(遗传漂变)或生境片段化促进了物种形成;复合群的不同物种间存在由基因渐渗引起的单倍型共享现象,可能是种间不完全的生殖隔离所致。花木蓝的居群单独聚为一枝,且与其他种无共享单倍型,因此应保留花木蓝种的分类地位,而其他种的分类地位有待进一步研究。     

西班牙桉树林下土小平盘菌复合群中两个独立的种——暗柔膜菌属一新组合及一新种:分类、分子生物学、生态与分布(英文)

土小平盘菌复合群原产澳大利亚,其中两个土生种在西班牙报道,很可能是在18世纪中叶引入西班牙。根据形态学特征和序列分析的结果,建立一个新组合和一个新种,分别定名为Phaeohelotium undulatum comb.nov.和Phaeoh.succineoguttulatum sp.nov.。对其宏观和微观形态特征进行了详尽的描述和图示,ITS rDNA序列分析的结果显示,它们以100%的支持率与Phaeohelotium monticola聚类在同一个分支。文中提供了该复合群Phaeohelotium属5个土生种的分种检索表,并对它们的生态习性与世界分布进行了深入讨论。     

紫乌头复合体nrDNA的ITS序列与系统发育分析

对乌头属 Aconitum L. 27个类群的nrDNA ITS序列进行了简约法与邻接法分析,两种方法得到的系统发育树基本一致。乌头亚属subgen. Aconitum 的蔓乌头系 ser. Volubilia 不是一个单系类群,显柱乌头系 ser. Stylosa 与兴安乌头系ser. Ambigua 各自作为单系类群也没有得到支持。特产于云南西北部横断山区的一些种之间存在非常近的系统发育关系,说明这些种可能是近期物种形成的产物。紫乌头 A. delavayi 复合体的不同类群在系统发育树上位     

Systematic study of basidiomycetous yeasts - evaluation of the ITS regions of rDNA to delimit species of the genus Rhodosporidium

Nucleotide sequences of internal transcribed spacer (ITS) regions were determined to establish the guidelines for species identification in the genus Rhodosporidium. Forty-two strains of nine species of the genus Rhodosporidium were used for ITS (ITS1 and ITS2) analysis. Intraspecific length polymorphisms and sequence variations were observed within R. azoricum, R. diobovatum, R. paludigenum, R. sphaerocarpum and R. toruloides, while no variation was observed within R. babjevae and R. kratochvilovae. Based on comparison of the levels of intraspecific and interspecific sequence similarity, strains with identical sequences were considered to represent a single species and strains with 92% or lower similarity of ITS sequences were considered to be distinct species in the genus Rhodosporidium.     

Characterization of Gibberella fujikuroi complex isolates by fumonisin B1 and B2 analysis and by RAPD and restriction analysis of PCR-amplified internal transcribed spacers of ribosomal DNA

Twenty nine isolates of Fusarium spp. (twenty four of them belonging to the Gibberella fujikuroi complex) isolated from banana and corn from different geographical regions were analyzed for their ability to produce fumonisins B1 and B2 and for genetic relatedness using random amplified polymorphic DNA (RAPD) and restriction analysis of PCR amplification products of the 5.8s ribosomal DNA-intervening internal transcribed spacer regions (ITS I-5.8S-ITS II). For RAPD analysis, six of twenty oligonucleotide primers were selected after testing with five Fusarium spp. isolates and used to characterize 24 additional isolates. DNA fragments from the 29 isolates of Fusarium spp., which were approximately 560 bp, were amplified with the universal primers ITS1 and ITS4. The restriction enzymes HaeIII, MboI, HpaII and MspI were useful for distinguishing the isolates. The RAPD analysis permitted to find interspecific differences among the isolates of Fusarium spp., between isolates with low and high capacity of fumonisin production and among isolates from different hosts. The restriction fragment length polymorphism (RFLP-PCR) analysis permitted to distinguish among different species of Fusarium. In combination with morphological analysis, the results of this research may find an application for the diagnosis of unknown Fusarium spp. and, particularly, for the characterization of fumonisin-producing isolates, which may be very useful in the food technology field.     

我国赫坎按蚊复合体成员种的rDNA-ITS2区序列差异及系统发育分析

测定了我国赫坎按蚊复合体 9成员种的核糖体DNA第二内转录间隔区 (rDNA ITS2 )序列 ,根据序列差异分析各蚊种间的系统发育关系。结果显示 :( 1 )ITS2区序列最长的是中华按蚊 ( 4 6 8bp) ,最短的是克劳按蚊和赫坎按蚊 ( 4 36bp) ;GC含量为 4 4 9%～ 4 6 8% ;( 2 )发现该复合体 4成员种的ITS2区序列存在种内个体间差异 ,幅度为 0～ 3 8% ,明显小于种间差异 ;( 3)将各蚊种的ITS2区序列进行同源排序比较 ,发现其变异大多是简单重复单元的拷贝数不同 ;种间差异性最大的是克劳按蚊与嗜人按蚊( 32 3% ) ,最小的是贵阳按蚊与凉山按蚊 ( 9 0 % )平均差异率为 2 2 3% ;( 4 )根据ITS2区序列特征 ,用 3种方法构建的树状图拟合一致。以上结果表明赫坎按蚊复合体各成员种rDNA ITS2序列在种内非常保守 ,以种间序列差异分析为基础的分子鉴别技术是甄别蚊种分类地位混淆和错误的有效方法。     

Phylogenetic relationships among the Caribbean members of the Cliona viridis complex (Porifera, Demospongiae, Hadromerida) using nuclear and mitochondrial DNA sequences

Species complexes - groups of closely related species in which intraspecific and interspecific variability overlap - have generated considerable interest and study. Frequently, members of a species complex do not have complete reproductive isolation; therefore, the complex may go through extensive gene flow. In the Caribbean Sea, some encrusting and excavating sponges of the genus Cliona (Porifera, Hadromerida, Clionaidae) are grouped within the great "Cliona viridis" complex because of their morphological similarities. This study examined the evolutionary relationships of the Caribbean members of this complex (C. caribbaea, C. tenuis, C. aprica and C. varians) and related taxa based on nuclear (ITS1 and ITS2) and mitochondrial (3' end of ND6) DNA sequences. The intragenomic ITS variation and its secondary structures were evaluated using a mixed approach of Denaturing Gradient Gel Electrophoresis (DGGE), DNA sequencing and secondary structure prediction. Considerable intragenomic variation was found in all the species, with apparently functional ITS1 and ITS2 secondary structures. Despite the subtle but clear morphological differentiation in these excavating sponges, the intragenomic copies of C. caribbaea, C. tenuis and C. aprica had a polyphyletic placement in the ITS1 and ITS2 genealogies and very low divergence. Therefore, it is clear that these species constitute a species complex (herein called Ct-complex). Genetic distances within the Ct-complex revealed that an important part of the interspecific variation overlapped with intraspecific variation, suggesting either incomplete lineage sorting or extensive gene flow. In contrast, C. varians and an unidentified "Pione" species emerged as monophyletic clades, being the closest sister groups to the Ct-complex. Additionally, our results support that C. laticavicola and C. delitrix conform a monophyletic group, but absence of reciprocal monophyly in these species suggests they may be life stages or ecophenotypes of a single species or they have diverged recently. Our work showed that the 3' end of the ND6 mitochondrial gene was highly conserved and not suitable for phylogenetic analysis at the interspecific level.     

Ribosomal DNA difference between species A and D of the Anopheles dims complex of mosquitoes from China

Abstract. Species A and D of the Anopheles dints complex were found in China. Ribosomal DNA second internal transcribed spacers (ITS2) of both species A and D were sequenced and found to be 716 and 710 base-pairs in length, respectively, with 699c GC content. No evidence of intraspecific variation was detected in the ITS2 sequence of species A, whereas the sequence of species D showed variation at one position in the ITS2. A large number of simple repeat motifs were dispersed throughout the ITS2 sequences. The level of interspecific difference was 5.4% of the nucleotide sequences. Some of the interspecific differences were located in regions with subrepeat structure.     

基于叶绿体DNA ndhA和nrDNA ITS序列变异分析栽培柿及其近缘种亲缘关系

以柿属植物（Diospyros spp.）中与柿近缘的8种共30个基因型为试材,进行核糖体DNA(nrDNA)内转录间隔区（ITS）和叶绿体DNA ndhA序列变异分析,并通过软件计算两个序列及合并后的进化模型,依据进化模型采用ML法(maximum likelihood method)分析进化关系。为进一步弄清柿属植物种间亲缘关系和供试柿（Diospyros kaki Thunb.）种内分子差异提供了理论依据。结果表明：（1）ndhA序列长度变异范围在1 492~1 511,14个信息位点;ITS序列长度变异范围在660~761,56个信息位点。ITS、ndhA和ndhA+ITS（ndhA和ITS合并）最适碱基进化模型分别为（TrN+I+G）、（F81+I）和（GTR+I+G）。综合ITS和ndhA序列分析表明：柿与油柿和云南野毛柿亲缘关系最近,与美洲柿和乌柿最远。（2）21份柿品种材料的ITS长度均为730,包括4个变异位点,据此4个变异位点对供试柿种内21个品种进行聚类分析。研究认为,ndhA和ITS能较清楚解释了柿与其近缘种间的亲缘关系,并通过柿品种ITS的差异位点分析鉴别出栽培柿种内的差异。     

The use of the polymerase chain reaction and restriction fragment length polymorphism technique associated with the classical morphology for characterization of Lymnaea columella, L. viatrix, and L. diaphana (Mollusca: Lymnaeidae)

The specific identification of Lymnaeid snails is based on a comparison of morphological characters of the shell, radula, renal and reproductive organs. However, the identification is complicated by dissection process, intra and interspecific similarity and variability of morphological characters. In the present study, polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) techniques targeted to the first and second internal transcribed spacers (ITS1 and ITS2) rDNA and to the mitochondrial 16S ribosomal gene (16S rDNAmt) were used to differentiate the species Lymnaea columella, L. viatrix, and L. diaphana from some localities of Brazil, Argentina, and Uruguay as well as to verify whether the molecular results corroborates the classical morphological method.PCR-RFLP analysis of the ITS1, ITS2, and 16S using 12 restriction enzymes revealed characteristic patterns for L. columella and L. diaphana which were concordant with the classical morphology. On the other hand, for L. viatrix populations a number of 1 to 6 profiles were generated while morphology provided the species pattern results.     

中国秋海棠属 (秋海棠科) 植物的DNA条形码评价

秋海棠属植物种类繁多,形态变异多样,导致种类的系统放置混乱,近缘种类鉴定困难。利用DNA条形码实现物种快速准确的鉴定技术具有不受形态特征约束的优势,为秋海棠属植物的分类鉴定提供了新的方法。本研究选择4个DNA条形码候选片段（rbcL,matK,trnH psbA,ITS）对中国秋海棠属26种136个个体进行了分析。结果显示：叶绿体基因rbcL,matK和trnH psbA种内和种间变异小,对秋海棠属植物的鉴别能力有限;ITS/ITS2种内和种间变异大,在本研究中物种正确鉴定率达到100%/96%,可考虑作为秋海棠属DNA条形码鉴定的候选片段。研究结果支持中国植物条形码研究组建议将核基因ITS/ITS2纳入种子植物DNA条形码核心片段中的观点。