小麦族披碱草属、鹅观草属和猬草属模式种的C带研究

采用改良的Giemsa C带技术,分析了小麦族披碱草属、鹅观草属和猬草属模式种的染色体C带带型。Elymus sibiricus、Roegneria caucasica和Hysrix patula的染色体在Giemsa C带带型上存在明显的差异,显示了这3个属模式种的物种特异性。3个模式种的Giemsa C带核型表明,C带带纹主要分布在染色体的末端和着丝粒附近,而中间带相对较少。对E.sibiricus、R.caucasica和H.patula的St、H、Y染色体组C带带型与其它物种的St、H、Y染色体组C带带型的差异进行了讨论。     

鹅观草属、披碱草属、猬草属和仲彬草属物种的RAMP分析及系统学意义

对鹅观草属、披碱草属、猬草属和仲彬草属4属23个物种进行了RAMP分析。结果表明属间变异极大,多态性极高。31个引物组合产生的286条DNA扩增片段均具有多态性。聚类分析显示鹅观草属、披碱草属、猬草属和仲彬草属物种各自聚为一类;Roegneria alashanica、R．elytrigioides和R．magnticaespes聚类在一起;猬草属的模式种Hystrix patula与披碱草属物种聚类在一起,而Hystrix duthiei和H．longearistata单独聚为一类;形态相似、染色体组相同、地理分布一致的物种聚类在一起。本研究结果基本上同形态学和细胞学研究结果相吻合,将鹅观草属、披碱草属和仲彬草属作为属分类等级处理比较恰当,而猬草属的系统地位有待进一步确认。RAMP标记可作为评价多年生小麦族物种遗传多样性和亲缘关系的一种分子标记技术。     

小麦族鹅观草属、披碱草属、猬草属和仲彬草属细胞质基因组PCR-RFLP分析

对鹅观草属、披碱草属、猬草属和仲彬草属23个物种和1份外类群共24份材料进行了细胞质基因组PCR-RFLP分析。3个叶绿体和3个线粒体通用引物扩增出的片段,用15种限制性内切酶对其进行酶切。在47种引物/酶组合中,获得329条DNA片段,其中304条具有多态性,占92.4%。结果表明鹅观草属、披碱草属、猬草属和仲彬草属材料存在属间和种间多态性,遗传相似系数较高。聚类分析显示仲彬草属单独聚为一类,鹅观草属R. grandis、R. aristiglumis、R. elytrigioides、R. alashanica、R. magnicaespes聚为一类,R. caucasica、R. ciliaris、R. amurensis、R. japonensis聚在一起,5个披碱草属材料、3个猬草属材料和1个鹅观草属物种R. kamoji聚为一类。这些结果与前人对其进行的RAPD和RAMP分析的结果基本一致。仲彬草属作为属分类等级处理是恰当的,对于鹅观草属、披碱草属和猬草属的系统地位和一些物种的分类处理,有待进一步研究。     

用RAPD分子标记探讨鹅观草属的种间关系

通过34个10碱基随机引物对鹅观草属(RoegneriaC.Koch)26个物种进行PCR扩增,28个引物能产生多态带。在NTSYS程序中,对16个引物扩增的186条扩增产物,计算Jaccard遗传相似系数,建立UPGMA聚类图。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)StY和StYH基因组的物种存在着一定的遗传差异,并各有一定程度的分化,这种分化是与地理位置相联系的。相距越远,物种相似程度越低;(3)形态差异较小,基因组同源,地理分布一致的物种聚类在一起,表现出较密切的亲缘关系;(4)无芒类群的R.alashanicaKeng和R.magnicaespes(D.F.Cui)L.B.Cai与其余分析的24个Roegneria物种存在着极大的遗传差异;(5)分布于西亚的R.caucasicaC.Koch.与分布于东亚和中亚的物种存在着较大的RAPD变异,亲缘关系较远;(6)在对R.ciliaris(Trin)Nevski和R.japonensis(Honda)Keng、R.tenuispicaJ.L.YangetY.H.Zhou和R.pendulinaNevski、R.tsukushiensis(Honda)Ohwi和R.kamojiOhwi等的分类处理上,基本上与形态学和细胞学的研究结果一致。对RAPD标记在Roegneria系统分类研究中的应用进行了讨论。     

小麦族四个属模式种的醇溶蛋白分析

利用酸性聚丙烯酰胺凝胶电泳 (APAGE)对小麦族披碱草属、鹅观草属、猬草属和仲彬草属 4个属的模式种进行了醇溶蛋白电泳分析 ,结果表明 :(1 ) 4个模式种具有明显的醇溶蛋白遗传多样性 ,其种间醇溶蛋白多态性高达 92 .3 % ;(2 ) Elymus sibiricus和 H ystrix patula具有相似的醇溶蛋白带型 ,而 Roegneria caucasica和Kengyilia gobicola的带型基本相似 ,其醇溶蛋白图谱能够反映一定的系统关系 ;(3 )不同收集地的 E. sibiricus材料间也存在明显的醇溶蛋白遗传差异 ,新疆的 E. sibiricus具有较丰富的醇溶蛋白带纹 ,而甘肃的 E. sibiricus的醇溶蛋白带纹较少。     

利用RAPD特异标记分析东北猬草染色体组成

选用5个染色体组特异的RAPD引物(St、H、Ns、Ee、Eb),对东北猬草[Hystrix komarovii (Roshev.) Ohwi]等5个猬草属及其8个近缘属物种进行PCR扩增,以探讨东北猬草的染色体组组成.结果显示:Hy.komarovii具有Ns染色体组特异的RAPD标记,而没有St、H、Ee和Eb特异的RAPD标记.表明Hy.komarovii含有Ns染色体组,而不含St和H染色体组,认为其染色体组组成可能与Hy.duthiei、Hy.coreana和Leymus arenarius一样,具有NsXm染色体组.根据染色体组分类原理,支持将东北猬草归于赖草属中.     

用RPAD分子标记探讨鹅观草属的种间关系

通过３４个１０碱基随机引物对鹅观草属（Ｒｏｇｅｇｎｅｒｉａ Ｃ．Ｋｏｃｈ）２６个物种进行ＰＣＲ扩增,２８个引物能产生多态带。在ＮＴＳＹＳ程序中,对１６个引物扩增的１８６条扩增产物,计算Ｊａｃｃａｒｄ遗传相似系数,建立了ＵＰＧＭＡ聚类图。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）ＳｔＹ和ＳｔＹＨ基因组的物种存在着一定的遗传差异,并各有在一定程度的分化,这种分化是地理位置相联系的     

石蒜属种间亲缘关系RAPD分析

采用RAPD标记构建了石蒜属Lycoris植物13个种的指纹图谱。从520个随机引物中筛选出清晰且多态性高的41个引物,共产生了350条DNA片段,分子量在0.3-3.0kh之间,其中253条谱带具有遗传多态性,约占72．3％,平均每个引物扩增的DNA片段数为6．2条。采用TFPGA数据分析软件,计算Nei氏相似性系数和遗传距离,建立了UPGMA聚类图。结果表明：石蒜属13个种明显聚为两大类,即具有单型核型结构（I型）,染色体基数为x=11的5个物种：玫瑰石蒜L.rosea,红蓝石蒜.L.haywardii,稻草石蒜L.straminea,换锦花L.sprengeri和石蒜L.radiata聚为一类;具有两型核型结构（V型和I型）8个物种即江苏石蒜L.houdyshelii,乳白石蒜L.albiflora,中国石蒜L.chinensis,长筒石蒜L.longituba,安徽石蒜Lanhuiensis,夏水仙L.squmigera,短蕊石蒜L.caldwellii和忽地笑L.aurea聚为一类。其中玫瑰石蒜和红蓝石蒜的亲缘关系最近,石蒜和忽地笑的亲缘关系较远,与细胞学的研究结果相吻合。通过RAPD分析,对玫瑰石蒜,红蓝石蒜和稻草石蒜是否天然杂交起源以及乳白石蒜,稻草石蒜和江苏石蒜的分类地位进行了探讨。     

10种披碱草属植物的RAPD分析及其系统学意义

利用随机扩增多态性DNA(RAPD)技术分析了10种披碱草属植物,即：Elymus sibiricus L.,E． caninus(L．)L¨E．lanceolatus(Scribner et Smith)Gould,E．nutans Griseb,E．dahuricus Turcz,, E．tangutorum(Nevski)Hand．-Mazz., E．brachyaristatus ALöve,E.submuticus(Keng)Keng f.,E． cylidricus(Franch)Honda和E．excelsus Turcz. 。对34个OPRON公司十聚体随机引物进行多态性筛 选,25个(75．53％)能产生多态性。16个引物产生的136个DNA片断,用于计算种间Nei氏遗传相似 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)四倍体和六倍体 物种各自聚为一支,它们之间的亲缘关系较远;(2)E．sibiricus和E. caninus;亲缘关系较近,支持把 Roegneria caninus (L．)Nevski归入Elymus;(3)E．nutans和E．dahuricus亲缘关系密切;(4)形态相 似、地理分布一致的E．brachyaristatus和E．submuticus存在着一定程度的核苷酸序列的差异,它们与 E．nutans和E．dahuricus有一定的亲缘关系;(5)E．excelsus 与 E．cylindricus的亲缘关系较近,它们 又与E．tangutorum有亲缘关系;(6)RAPD结果与形态学和细胞学等分析结果基本一致,RAPD分析方法将为Elymus系统分类提供DNA水平上丰富的资料。     

大鹅观草与蛇河披碱草属间杂种的细胞遗传学研究

为探讨大鹅观草(Roegneria grandis,2n=4x=28)的染色体组组成,为其正确的分类处理提供细胞学依据。该研究通过人工远缘杂交,成功获得3株大鹅观草与蛇河披碱草(Elymus wawawaiensis,2n=4x=28)属间杂种F1植株。杂种植株形态介于两亲本之间,不育。亲本及杂种经I2-IK溶液染色后进行花粉育性检测,结果显示Roegneria grandis和Elymus wawawaiensis的花粉可育,育性高达94.6%和90.5%;杂种F1不育。花粉母细胞减数分裂中期I染色体配对结果显示,亲本花粉母细胞配对正常,均形成14个二价体,以环状二价体为主,Roegneria grandis有频率很低(0.04/细胞)的单价体出现;杂种F1平均每个花粉母细胞形成6.46个二价体,变化范围为5~8;在观察的83个花粉母细胞中,有35.2%的花粉母细胞形成了7个二价体,形成6个二价体的细胞占42.59%,较少细胞形成8个二价体;平均每个细胞形成14.66个单价体,变化范围为10~18;平均每细胞观察到0.14个三价体;杂种花粉母细胞染色体构型为14.66 I+6.46 II+0.14 III;平均每细胞交叉数为9.83,C值为0.35。结果表明:(1)R.grandis与Elymus wawawaiensis有一组染色体组同源的St染色体组,另外一组染色体组不是St或者H染色体组,Roegneria grandis的染色体组组成不是St Stg;(2)较低频率的三价体(平均0.14个/细胞),可能是由于R.grandis的St和Y染色体组间具有一定的同源性,也可能是染色体易位等原因导致,对于Y染色体组的起源还需深入地研究;(3)在不同地理来源的披碱草属和鹅观草属物种中St染色体组同源性不同,R.grandis与来自于北美的Elymus lanceolatus与E.wawawaiensis的St染色体组较与分布于亚洲的E.sibiricus和E.caninus的St染色体组同源性反而更高,其原因还需要进一步地研究。     

RAPD Study on Inter species Relationships in Roegneria (Poaceae: Triticeae)

To assess the relationships among 26 species in Roegneria C. Koch, 34 random decamer primers were screened for RAPD fragments. 28 primers produced polymorphic RAPD products. Data from 16 primers were used for RAPD assay. By NTSYS-pc program, Jaccard' s genetic similarity coefficients were generated and dendrogram was constructed using UPGMA. It is concluded as follows: (1) Distinct genetic differences and extensive genetic diversity were present among the species. (2) There were some genetic differences between StY and StYH genomes, and StY and StYH had a certain degree of differentiations respectively which were related to geographic regions, the farther the geographic distribution between species, the less the similarity to each other. (3) When different accessions in a species, such as species with similar morphological characters, homologous genomes and similar geographic distribution, were clustered together respectively, it suggusted that they had closer relationships. (4) The awnless species R. alashanica Keng and R. magnicaespes (D. F. Cui) L. B. Cai, in Roegneria, were separated from the other species analysed in this study, indicating that these two species had intensive genetic differences from the others. (5) R. caucasica C. Koch, a species from Western Asia, was quite different from the other species contained StY genomes in Roegneria from Eastern Asia and Central Asia. (6) The results were in consistance with that of the analysis of morphology and chromosome pairing in the taxonomic treatments for R. ciliaris (Trin) Nevski and R. japonensis (Honda) Keng, R. tenuispica J. L. Yang et Y. H. Zhou and R. pendulina Nevski, and R. tsukushiensis (Honda) Ohwi and R. kamoji Ohwi. The present study discussed the usefulness of RAPD markers in the systematic study of Roegneria.     

披碱草属(Elymus L.)两个物种之订正

披碱草属Elymus L．是小麦族Triticeae Dumort．中最大的属,全世界共有150多个种,广泛分布于 温带地区。我国约有80余种(包括鹅观草属Roegneria C．Koch),是披碱草属的重要分布区和多样性分 化中心。由于披碱草属植物种类繁多、分布广泛、生境多样以及形态变异较为复杂,导致该属在分类上 存在许多问题。同时,由于地域的局限和地区间交流的缺乏,致使同一种披碱草属植物被不同的学者多 次发表及同物异名现象的出现。本文对E．antiquus和E．burchan-buddae这两个曾多次被订名的披碱草 属物种进行了分类订正。     

Phylogeny of Hystrix and related genera (Poaceae: Triticeae) based on nuclear rDNA ITS sequences

The taxonomic status of Hystrix and phylogenetic relationships among Hystrix and its related genera of Pseudoroegneria (St), Hordeum (H), Psathyrostachys (Ns), Elymus (StH), Leymus (NsXm), Thinopyrum bessarabicum (E(b)) and Lophopyrum elongatum (E(e)) were estimated from sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA. The type species of Hystrix, H. patula, clustered with species of Pseudoroegneria, Hordeum, Elymus, Th. bessarabicum and Lo. elongatum, while H. duthiei ssp. duthiei, H. duthiei ssp. longearistata, H. coreana and H. komarovii were grouped with Psathyrostachys and Leymus species. The results indicate that: (i) H. patula is distantly related to other species of Hystrix, but is closely related to Elymus species; (ii) H. duthiei ssp. duthiei, H. duthiei ssp. longearistata, H. coreana and H. komarovii have a close affinity with Psathyrostachys and Leymus species, and H. komarovii might contain the NsXm genome of Leymus; and (iii) the St, H and Ns genomes in Hystrix originate from Pseudoroegneria, Hordeum and Psathyrostachys, respectively, while the Xm in Hystrix and Leymus has a complex relationship with the E or St genomes. According to the genomic system of classification in Tiritceae, it is reasonable to treat Hystrix patula as Elymus hystrix L, and the other species of Hystrix as species of a section of Leymus, Leymus Sect. Hystrix.     

八个四倍体鹅观草属物种的核型研究

对8个鹅观草属物种的核型进行了研究。核型公式如下:纤穗鹅观草2n=4x=28=20m+8sm(2sat);紫穗鹅观草2n=4x=28=22m(2sat)+6sm;假花鳞草2n=4x=28=24m(2sat)+4sm;肃草2n=4x=28=22m+6sm(2sat);小颖鹅观草2n=4x=28=22m+6sm(2sat);纤瘦鹅观草2n=4x=28=24m(4sat)+4sm;变颖鹅观草2n=4x=28=20m+8sm(2sat);毛花鹅观草2n=4x=28=24m(2sat)+4sm。它们的核型属1A或2A型。其中后5个物种的核型为首次报道。     

Genome analysis of species in the genus Hystrix (Triticeae; Poaceae)

Genomic in situ hybridisation (GISH) and Southern genomic hybridisation were applied in order to gain further knowledge regarding generic delimitation of the genus Hystrix as well as to clarify the genomes of the Hystrix species H. patula, H. longearistata, H. coreana, H. duthiei and H. komarovii. The hybridisation intensity of different genomic probes was compared among the Hystrix species and with other Triticeae species. The Southern- and GISH results confirm that H. patula contains the StH genome and show that H. komarovii most likely has a variant of this StH genome. The other Hystrix species under study, i.e. H. longearistata, H. coreana and H. duthiei, contain an Ns basic genome, and most probably two variants of this basic genome, Ns ¹ Ns ² . The genus Hystrix is thus not a monophyletic group of species.     

用RAPD标记研究蚱属五个种间的亲缘关系

用RAPD技术对蚱属5种蚱基因组DNA的多态性进行研究。在事先优化的反应条件下用12个随机引物扩增, 共得到84条清晰稳定的多态性片段,片段长度为200～2 000 bp。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。