利用RAPD标记构建木耳属种间关系的研究

利用RAPD标记以银耳属的2个种为外类群研究了木耳属8个种的分子系统发育关系,根据UPGMA构建的树状图结果表明木耳属（Auricularia)和银耳属（Tremella)分别为单一的分类单元,木耳属的8个种被分成3个明显不同的组,第一组包括循形木耳（A.peltata Lloyd)和角质木耳（A.cornes Spreng.),第二组包括所有供试的黑木耳（A.auricula Underw.),第三组包括皱木耳（A.delicata Henn.)、琥珀木耳（A.fuscosuccinea Farl.)、毛木耳（A.polytricha Sacc.)、大木耳（A.maximaY.R.Guo)和网膜木耳（A.reticulataL.J.Li)五个形态种,种间发育关系表明角质木耳和毛木耳应是两个不同的种而非同种异名,子实体横切面中髓层的有无并不表明不同种间亲缘关系的远近。     

利用RAPD技术对木耳属菌株进行分类鉴定的研究

利用ＲＡＰＤ技术对木耳属不同种和种内不同菌株进行了分子鉴定。结果表明,在供试的２６个随机引物中,有１０个引物可扩增出清晰、稳定的ＤＮＡ带型,其中引物Ｓ４和Ｓ６与供试木耳属菌株的基因组ＤＮＡ结合位点少而保守,扩增图谱具有种的特异性,可用于种的快速准确鉴定;其综随机引物的扩增ＤＮＡ多态性较强,可用于种内不同菌株的鉴定;聚类分析表明,在７５％相似水平下,可将供试的木耳属菌株分成四大类,其中有三大类各代表一个形态鉴定的种。研究结果表明了ＲＡＰＤ技术可有效地用于木耳属种或菌株的快速准确鉴定。     

利用RAPD技术对木耳属菌株进行分类鉴定的研究

利用ＲＡＰＤ技术对木耳属不同种和种内不同菌株进行了分子鉴定。结果表明,在供试的２６个随机引物中,有１０个引物可扩增出清晰、稳定的ＤＮＡ带型,其中引物Ｓ４和Ｓ６与供试木耳属菌株的基因组ＤＮＡ结合位点少而保守,扩增图谱具有种的特异性,可用于种的快速准确鉴定;其综随机引物的扩增ＤＮＡ多态性较强,可用于种内不同菌株的鉴定;聚类分析表明,在７５％相似水平下,可将供试的木耳属菌株分成四大类,其中有三大类各代表     

木耳属种间杂交技术研究 Ⅰ.木耳菌丝原生质体的形成与再生

本文比较了不同酶液、渗透压稳定剂、酶解温室及菌丝培养基成份等因素对木耳属(Auricularia)中木耳(Auricularia auricula)和毛木耳(Auricularia polytricha)菌丝释放原生质体的作用及影响。用0.5%纤维素酶加0.5%蜗牛酶的混合酶液,以0.6M的MgSO_4为稳定剂,在34℃下可自两种菌丝体获得大量原生质体。对原生质体再生条件的研究表明,纤维二糖和菌丝体培养物浸提物对再生有明显促进作用,再生率达20%左右。本文还用VBL型荧光增白剂观察了菌丝脱壁以及原生质体细胞壁再生的过程。     

利用RAPD技术对木耳属菌株进行分类鉴定的研究*

利用ＲＡＰＤ技术对木耳属不同种和种内不同菌株进行了分子鉴定。结果表明,在供试的２６个随机引物中,有１０个引物可扩增出清晰、稳定的ＤＮＡ带型,其中引物Ｓ４和Ｓ６与供试木耳属菌株的基因组ＤＮＡ结合位点少而保守,扩增图谱具有种的特异性,可用于种的快速准确鉴定;其综随机引物的扩增ＤＮＡ多态性较强,可用于种内不同菌株的鉴定;聚类分析表明,在７５％相似水平下,可将供试的木耳属菌株分成四大类,其中有三大类各代表一个形态鉴定的种。研究结果表明了ＲＡＰＤ技术可有效地用于木耳属种或菌株的快速准确鉴定。     

用RAPD分子标记探讨沙拐枣属的种间关系

利用随机扩增多态性DNA（RAPD）技术分析了14种沙拐枣属（Calligonum L.)植物,通过对16个Sangon公司十聚体随机引物进行PCR扩增,3个引物能产生多态性带。对3个引物扩增产生的45条扩增产物,计算单匹配系数,应用UPGMA方法构建亲缘关系树状图。分析结果表明：（1）物种间遗传差异明显,具有丰富的遗传多样性;（2）14种沙拐枣属植物明显聚为4类,与传统的形态学分类结果基本一致。     

海南岛木耳属的研究

子实体单生,鲜时坚韧的胶质,不透光,耳状,1-2×2.5-3.5cm,厚2-3.1mm,基部发达,呈黄褐色,不孕面有短柔毛和许多短的裂缝,栗褐色,干时棕黑色;子实层有粗条状隆起密集的结节,紫红色,干后紫黑色。柔毛层:毛长30-80(-100)μm,粗4-5.5μm,无色到褐黄色,不成簇,具狭窄的腔,顶园,易截断,基部明显膨大或再突然收缩成细而长的根状;致密层:20-50μm     

篮状菌属的四个中国新记录种

在系统调查我国篮状菌属Talaromyces物种资源的工作中,分离得到了10株篮状菌.基于形态学和BenA、CaM及rDNA ITS1-5.8S-ITS2序列的分子种系学分析确定它们分属于6个种,即艾米斯托克篮状菌T.amestolkiae、暗玫瑰篮状菌T.atroroseus、暗绿篮状菌T.fuscoviridis、...     

利用RAPD分子标记评价仲彬草属的种间关系

利用随机扩增多态性DNA(RAPD)技术分析了14种仲彬草属Kengyilia植物的种间关系。对34个OPRON公司十聚体随机引物进行多态性筛选,20个(58．8％)能产生多态性。14个引物产生的112个DNA片断,用于计算种间Jaccard遗传相似性系数分析,在NTSYS程序中利用UPGMA构建系统发育树状图。分析结果表明：(1)14个Kengyilia物种存在较大的遗传多样性;(2)青藏高原的物种与新疆的物种 的RAPD变异极大;(3)形态相似、地理分布一致的物种有一定的亲缘关系,聚类在一起;(4)RAPD结果与形态学和细胞学等分析结果一致。RAPD分析方法将为Kengyilia系统分类提供DNA水平上丰富的资料。     

光木耳和琥珀木耳种间营养缺陷型原生质体融合研究

利用~(60)Co—γ射线辐照木耳担孢子,获得了9株营养缺陷型突变体。采用光木耳[Auricularia auricula(L,ex Hook.)Underw.]组氨酸缺陷型菌株Aa-γH-9和琥珀木耳[Auricularia fuscosucinea(Mont.)Farlow]腺嘌呤缺陷型菌株Af-γH-1进行种间原生质体融合实验,根据营养互补原理,在基础培养基(MM)上检出融合子,获得了稳定的融合子,融合子频率为3.34×10~(-4)—3.76×10~(-4)。初步遗传分析表明:融合子细胞核为单核,是单核异核体,融合子氨基酸含量、酯酶同功酶酶谱均与双亲不同。     

MOLECULAR PHYLOGENY OF GRACILARIA SPECIES INFERRED FROM MOLECULAR MARKERS BELONGING TO THREE DIFFERENT GENOMES1

The nucleotide sequence data of molecular markers 18S rRNA, RUBISCO spacer, and cox2‐3 intergenic spacer were integrated to infer the phylogeny of Gracilaria species, collected from the western coast of India, reducing the possibility of misidentification and providing greater phylogenetic resolution. A phylogenetic tree was constructed using cox2‐3 and RUBISCO spacer sequences, exhibiting the same clustering but differing slightly from that of the rRNA‐based phylogenetic tree. The phylogeny inferred from the combined data set confers an analogous pattern of clustering, compared with those of trees constructed from individual data sets. The combined data set resulted in a phylogeny with better resolution, which supported the clade with higher consistency index, retention index, and bootstrap values. It was observed that Gracilaria foliifera (Forssk.) Børgesen is closer to G. corticata (J. Agardh) J. Agardh varieties, while G. salicornia (C. Agardh) E. Y. Dawson and G. fergusonii J. Agardh both originated from the same clade. The position of G. textorii (Suringar) De Toni faltered and toppled between G. salicornia and G. dura (C. Agardh) J. Agardh; however, G. gracilis (Stackh.) M. Steentoft, L. M. Irvine et W. F. Farnham was evidently distant from the rest of the species.     

五种缘毛类纤毛虫遗传关系的ISSR研究

应用ISSR分子标记揭示了5种缘毛类纤毛虫(Carchesium polypinum,Epistylis chrysemydis,E.plicatilis,E.urceolata和Vorticella campanula)的遗传关系.从34个引物中筛选到13个多态性高的引物进行研究.得到的遗传距离(0.666 7～1.000 0)显示ISSR技术具有较高的分辨率.依据构建的UPGMA聚类树,V.campanula首先和其他种类分开;C.polypinum和E.chrysemydis聚在了一起;在3种Epistylis纤毛虫中,E.plicatilis和E.urceolata聚在了一起.对比由核糖体小亚基RNA基因序列构建的系统发育树,发现:1)ISSR引物在缘毛类纤毛虫基因组中可以得到多态扩增;2)C.polypinum与Epistylis的关系近于V.campanula,在缘毛类纤毛虫分类中,单生或群居是重要的系统发育特征;3)E.plicatilis和E.urceolata的关系近于E.chrysemydis.在Epistylis属纤毛虫中,柄的中空与否是一个有用的系统发育和分类特征.本研究表明ISSR方法在纤毛虫相近和相似种遗传关系研究中是一种新的有用方法.     

PHYLOGENETIC RELATIONSHIPS AMONG ISOLATES OF EUDORINA SPECIES (VOLVOCALES, CHLOROPHYTA) INFERRED FROM MOLECULAR AND BIOCHEMICAL DATA

Phylogenetic analyses of 19 strains representing five species of Eudorina, one strain of Pleodorina indica, and seven strains of Yamagishiella unicocca were carried out by sequencing the internal transcribed spacer region (ITS 1 and ITS 2) of the nuclear ribosomal DNA (rDNA) repeats. The sequence data resolved five phylogenetic groups, one consisting of Y. unicocca and the other four encompassing all the Eudorina species. Two isolates, Eudorina sp. (ASW 05157) and Pleodorina indica (ASW 05153), were of uncertain affiliation. Whereas one monophyletic group included strains of E. elegans only, the other strains of E. elegans appeared alongside E. cylindrica, E. illinoisensis, and E. unicocca var. unicocca in the other Eudorina clades. The distribution pattern of the carotenoid loroxanthin ([3R,3'R,6'R]-β,ε-carotene-3,19,3'-triol), a systematically useful biochemical marker within chlorophycean flagellates, was shown to match the evaluated molecular data. Whereas it was either totally absent or universally present in six of the deduced phylogenetic lines, it occurred randomly in the E. elegans clade containing only E. elegans isolates. The results substantiated the current hypothesis that the unique vegetative morphology of E. elegans has independently arisen at various times during evolution and that it is not a marker of a monophyletic group.     

用RAPD标记研究对虾属六个种间的亲缘关系

用ＲＡＰＤ技术对对虾属中六种对虾基因组ＤＮＡ的多态性进行研究。在事先优化的反应条件下用２０个随机引物扩增,共得到３６４条清晰稳定的多态性片段,片段长度在２３０￣２８００ｂｐ之间。统计这些片段,根据扩增片段的共享度计算出相对遗传距离指数,然后用ＵＰＧＭＡ和ＮＪ等聚类方法对其进行分析,构建系统树,确定了它们相互间的亲缘关系。从聚类分析的结果可以看到,外部形态比较相似的种类在基因组ＤＮＡ上表现出较大的相     

TAXONOMIC REVISION OF SARGASSUM SPECIES (FUCALES,PHAEOPHYCEAE) FROM NEW CALEDONIA BASED ON MORPHOLOGICAL AND MOLECULAR ANALYSES1

Sargassum C. Agardh (1820) is a taxonomically difficult genus distributed worldwide and reported as the most species‐rich genus of the Fucales. It is especially abundant in the Pacific where decreasing species richness is reported to occur from west to east. New Caledonia has been recognized as one of the hotspots of Sargassum diversity; however, species lists available for this region are old and incomplete and have not yet been updated with regard to the latest taxonomic revisions published. This study aimed at revising Sargassum diversity in New Caledonia and to assess its geographic affinities with neighboring Pacific regions. We used combined morphological and DNA analyses on new collections and examined numerous type specimens. Although 45 taxa have been listed in the literature, most of them have been either transferred to synonymy since or misidentified, and in this study, only 12 taxa were recognized as occurring in New Caledonia. They belong to the subgenus Sargassum sect. Binderianae (Grunow) Mattio et Payri (2), sect. Ilicifoliae (J. Agardh) Mattio et Payri (2), sect. Polycystae Mattio et Payri. (1), sect. Sargassum (4), sect. Zygocarpicae (J. Agardh) Setch. (2), and subgenus Phyllotrichia (Aresh.) J. Agardh (1). New Caledonian Sargassum flora appeared as the second richest in the region after the Pacific coast of Australia, with which it has shown high similarity, and shared species with all neighboring regions. One species, S. turbinarioides Grunow, is considered as endemic to New Caledonia. The low genetic diversity detected among several polymorphic species belonging to sect. Sargassum is also discussed.     

木耳属二新种

本文报道了在西沙群岛采集的木耳属二新种,命名为西沙木耳和网脉木耳,并附汉文和拉丁文描述。此外,还讨论了新种与近似种之间的区别。     

五株纤毛虫遗传关系的ISSR分析

采用ISSR分子标记技术,尝试对四种五株纤毛虫(褶累枝虫(Epistylis plicatilis)、绿草履虫(Paramecium bursaria)、多态喇叭虫(Stentor polymorphus)、嗜热四膜虫BF1株(Tetrahymena thermophilaBF1)和嗜热四膜虫BF5株(T.ther-mophilaBF5))进行遗传关系研究。用13个ISSR引物对五株纤毛虫进行扩增,六个ISSR引物获得多态片段。根据Nei s遗传距离矩阵构建了五株纤毛虫的遗传关系树状图。UPGMA,NJ聚类图表明:两株嗜热四膜虫最先聚在一起;其次是褶累枝虫和多态喇叭虫聚在一起,然后再与嗜热四膜虫聚在一起;咽膜亚纲的绿草履虫形成独立的一枝。结果显示:①缘毛亚纲纤毛虫可能是寡膜纲中较独特的一个类群,建议提升缘毛亚纲纤毛虫的分类地位;②缘毛亚纲褶累枝虫与膜口亚纲嗜热四膜虫的亲缘关系近于咽膜亚纲绿草履虫,在寡膜纲中绿草履虫处于原始地位;③五株纤毛虫基因组中均含有微卫星DNA序列:(GTG)4(、GACA)4(、AG)8(、CAA)6和(GAA)6。     

基于rDNA ITS序列探讨部分腐霉种的系统发育与其形态特征

基于对73株计58种腐霉和6种疫霉的核糖体DNA的ITS序列分析,以海生疫霉为外围群,按邻接法构建系统发育树,对腐霉的系统发育关系进行了研究。结果表明:在58种腐霉中,Pythium ostracodes,P.chamaehyphon,P.carbonicum,P.montanum和P.vexans归为同一组,介于其它腐霉和疫霉之间,这5种腐霉的孢子囊均为球形;现已归为疫霉属的P.undulatum 单独为一组,它与腐霉的亲缘关系比疫霉更近;其余52种腐霉聚成一大组,这52种腐霉基本上按孢子囊或菌丝膨大体形态分成Ⅰ、Ⅱ两组:第1组31种腐霉, 其中30种腐霉的孢子囊或菌丝膨大体为球形;第Ⅱ组21种腐霉,其中19种腐霉的孢子囊为丝状、瓣状或裂片状。基于ITS序列分析,腐霉属的其它性状如藏卵器壁是否光滑、卵孢子是否满器、雄器的着生方式和数量、异宗配合等呈多元演化。     

木耳属二新种*

本文报道了在西沙群岛采集的木耳属二新种,命名为西沙木耳和网脉木耳,并附汉文和拉丁文描述。此外,还讨论了新种与近似种之间的区别。     

基于16S rDNA序列探讨十种臂尾轮虫的系统关系和分类地位

通过对角突臂尾轮虫、尾突臂尾轮虫、裂足臂尾轮虫、剪形臂尾轮虫、方形臂尾轮虫、壶状臂尾轮虫、红臂尾轮虫、镰肜臂尾轮虫、萼花臂尾轮虫和十指臂尾轮虫等十种臂尾轮虫和大肚须足轮虫的线粒体16s rDNA进行扩增和序列测序,结合Genebank 中十指臂尾轮虫的16S rDNA序列,使用MAGE软件构建了NJ(neighbor-joining method)树,使用mrbayes软件构建了贝叶斯树,探讨了十种臂尾轮虫之间的系统关系.结果表明,本研究所涉及的轮虫16S rDNA序列差异百分比均值为14.6%,可作为分子标记应用于轮虫属内种间系统关系研究;系统树均支持将十指臂尾轮虫、裂足臂尾轮虫隶属于臂尾轮属;壶状臂尾轮虫和红臂尾轮虫是两个独立的种.此外,还依据16S rDNA序列变异百分比推测了十种臂尾轮虫的分化时间.