用核糖体DNA的ITS序列探讨裂叶沙参的系统位置——兼论ITS片段在…

本采用ＰＣＲ直接测序方法,对来自沙参属全部２组７亚组的１０个种和作为外类群的风铃草属２个种的核糖体ＤＮＡ ＩＴＳ片段进行了序列分析。在重点探讨裂叶沙参分类地位的同时,分析了ＩＴＳ片段序列的沙参属系统发育重建中的价值。结果表明,在沙参属中,ＩＴＳ片段在长度、ＧＣ含量和位点变异量上均比较一致;长度５３９ｂｐ￣５４１ｂｐ,ＧＣ含量５７％￣６０％,信息位点与只占总位点的３．９％。采用ＰＡＵＰ软件进行的系     

濒危物种裂叶沙参及其近缘广布种泡沙参的遗传多样性研究

根据对１２个形态性状的统计分析和１０个基因位点的等位酶检测,探讨了濒危植物裂叶沙参及其近缘广布种泡沙参的遗传多样性水平,对３个裂叶沙参和６个泡沙参天然群体的遗传分析表明,两种沙参属植物均具有很高的遗传变异水平,这种变异性既体现在形态学水平上,也体现在酶位点水平上,在２个茎叶形状以及１０个花果和种子形状上,濒危种裂叶沙参的变异性均与广布种相当,同样,根据７个酶系统１０个等位酶位点的度量,裂叶沙参群体     

根据ITS序列证据重建防己科蝙蝠葛族的系统发育

研究了国产防己科蝙蝠葛族tirb．Menispermeae9属20种和外类群青牛胆族trib．Tinosporeae 2属3种植物完整的ITS(包括5.8S rDNA)序列。trib．Menispermeae的ITS长527～601 bp,排序后长667bp。当gap处理为missing时具281个有信息位点。PAUP软件分析结果表明：①trib．Menispermeae是一个单系类群,该分支得到hootstrap l00％的支持;②确定了存疑种Pachygone valida的系统学位置,该种是Coc—culus属的成员;③Sinomenium和Menispermum两属有很近的系统学关系,组成族内稳定的一支,它们的ITS序列同源性极高,ITS1比族内其它属长41～73bp;④Stephania和Cyclea也是系统发育关系很近的两个类群。前者具两个主要分支,其IIS1、ITS2的G+C含量差异较大,在种类组成上,该两大支与传统上Stephania属内处理的2个亚属——千金藤亚属subgen．Stephania和山乌龟亚属subgen．Tuberiphania基本一致;Cyclea属内种间的ITS序列差异小,同源性极高。     

裂叶沙参与泡沙参种群分布格局分形特征的分析

 裂叶沙参与泡沙参种群在不同海拔区域分布格局的计盒维数与其在群落中占据空间的能力成正比,并且与环境因素密切相关。在裂叶沙参与泡沙参种群较为适生的区域,海拔2700～3100m,两者在群落中占据空间的能力最强,他们的计盒维数达到最高。在低于海拔2700m或高于海拔3100m,环境条件严酷的区域,裂叶沙参种群与泡沙参种群占据空间的能力不同程度的下降。在不同的海拔区域,广布种泡沙参种群占据空间的能力均比裂叶沙参种群高,而其受环境因素影响较弱。在海拔2700m以下地区,泡沙参种群水平分布格局的计盒维数比裂叶沙参种群高4.5倍;在两种群较为适生的区域,海拔2700～3100m,泡沙参计盒维数比裂叶沙参种群高1.5倍,差异最小;在泡沙参与裂叶沙参种群分布的上限,海拔3100m以上地区,两者计盒维数差异又增加,泡沙参计盒维数比裂叶沙参种群高1.8倍。这说明裂叶沙参种群的内在适应力、竞争力比泡沙参种群相对较弱。     

濒危种裂叶沙参种群生命表和存活曲线的研究及其与广布种泡沙参的对照

以近缘广布种泡沙参为对照,本文编制了濒危种裂叶沙参种群的标准生命表、存活曲线以及海拔２３００～２４００ｍ、２７００～２９００ｍ、２９００～３１００ｍ和３１００～３４００ｍ区域４个种群的生命表,存活曲线。根据其生活史,讨论了裂叶沙参各种群特定年龄的存活量、死亡量和死亡率等重要参数。裂叶沙参种群存活曲线属ＤｅｖｅｙＣ型,１～５年生死亡率最高,个体最长寿命２５年。而泡沙参种群的死亡高峰期是１～３年,个体最长寿命２１年。导致裂叶沙参种群幼龄个体死亡的内在原因是其较弱的抗逆性和适应能力;外在原因是严重的干旱、外界干扰和群落内的光照不足。裂叶沙参种群适应不良环境的能力较泡沙参弱：在海拔２７００ｍ以下地区,由于外界干扰和环境干旱,除水份条件较好的特殊生境,大多种群呈严重衰退趋势,不能满足编表条件,只有海拔２７００ｍ以上区域的种群呈稳定或扩展趋势;而泡沙参各种群在不同海拔区域均呈稳定或扩展趋势,满足编表条件。从海拔２３００ｍ到３４００ｍ,不同裂叶沙参种群个体寿命呈：短长短的格式,与外界严酷较好严酷的环境相对应。这表明裂叶沙参种群在严酷环境条件下改变了其生存对策,缩短了个体寿命。     

不同产地瓠瓜品种ITS序列的遗传多样性分析

对国产29个瓠瓜也Lagenariasiceraria（Molina）Standl.页品种的ITS序列进行了扩增及测序,并结合引自GenBank的国产9个瓠瓜品种以及国外6个瓠瓜品种和3个同属种类的ITS序列,对它们的ITS序列长度和GC含量以及变异位点进行比较,在此基础上构建系统发育树并对47个样本间的遗传关系进行研究。结果显示：供试47个样本的ITS序列均由ITS1、5.8SrDNA及ITS2组成,各样本间的ITS序列长度、GC含量以及变异位点差异明显。国产38个瓠瓜品种的ITS序列（包括ITS1、5.8SrDNA及ITS2）长度为619-627bp、GC含量为58.00%-63.32%;国外9个样本的ITS序列长度为591-626bp,GC含量为54.17%-63.26%。序列比对结果显示：国产38个瓠瓜品种的ITS序列同源率为84.6%-100.0%,包含221个变异位点;其中,来源于山东的品种‘砧木2’（‘ZhenmuNo.2’）的ITS序列包含的变异位点最多,与其他品种间的同源率也最低。在系统发育树上,国产38个瓠瓜品种可分为3个分支,来源于山东的品种‘砧木2’和来源于河南的品种‘西瓜砧木1’（‘XiguazhenmuNo.1’）各自聚为第1和第2分支;其余36个品种聚为第3分支。而供试的47个样本则可分为2个分支和5个亚组,第1分支可分为2个亚组,包括国产品种‘砧木2’和产自日本的2个品种;第2分支包含的44个样本则进一步分为3个亚组,国产品种‘西瓜砧木1’和产自法国的品种‘白花瓠瓜’（‘White-floweredgourd’）各自聚为第1和第2亚组,其余的42个样本聚为第3亚组。研究结果表明：供试的不同产地瓠瓜品种间存在丰富的遗传变异和地理分化现象,其ITS序列差异与地理分布有一定关系。     

裂叶沙参和泡沙参种群有性生殖和无性繁殖的比较

通过对裂叶沙参(Adenophora lobophylla Hong)和泡沙参(A.potaninii Korsh.)种群有性生殖和无性繁殖特性的对比,揭示濒危植物裂叶沙参的种群内部致濒机制。结果表明:裂叶沙参种群幼苗抢占草本层上层空间能力弱于泡沙参种群;裂叶沙参种群是以相对长的生殖期和高产籽量来适应环境;裂叶沙参种群开花结实量虽高于泡沙参种群,但其中成熟种子少,种子质量差,致使其种群由种子到一年生幼苗的转化率极低,并且幼苗生活力弱,是导致其种群濒危的重要内部原因。裂叶沙参种群除有性生殖外还兼有较弱的无性繁殖,是对其有性生殖的一个重要补充。     

基于ITS序列的不同产地裂叶荆芥系统发育分析

采用PCR法扩增来自国内5个不同产地的裂叶荆芥ITS全序列,测序后以产自河北的裂叶荆芥代表中国裂叶荆芥与来自不同国家的裂叶荆芥ITS序列进行比较,构建分子系统发育树,探讨国内5个不同产地及不同国家的裂叶荆芥的亲缘关系和系统进化。结果表明:国内5个不同产地的裂叶荆芥ITS1和ITS2序列均有较高的G/C含量;5个产地的裂叶荆芥的扩增序列长度均为749bp,且序列完全相同;其中ITS1序列长231bp,5.8S序列长168bp,ITS2序列长236bp。中国裂叶荆芥与日本、韩国裂叶荆芥ITS序列一致性为100%,与美国裂叶荆芥ITS序列一致性为99.0%。与美国裂叶荆芥相比,中国裂叶荆芥ITS序列有7个碱基发生变异。来自不同国家的裂叶荆芥形成单系群和2个分支(中、日、韩3国为1个分支,美国单独形成1个分支)。ITS序列的一致性表明国内5个不同产地裂叶荆芥为同一个种。     

遮光条件下裂叶沙参和泡沙参气孔行为的对比研究

通过遮光实验研究了裂叶沙参（Ａｄｅｎｏｐｈｏｒａ ｌｏｂｏｐｈｙｌｌａ）和泡沙参（Ａ．ｐｏｔａｎｉｎｉｉ）叶片的气孔行为,比较了不同遮光处理对裂叶沙参和泡沙参气孔的生理特性以及生态适应的影响。结果表明：裂叶沙参对不同遮光处理的适应性较差,相反,泡沙参具有较强的适应性和抗逆性;在自然光照和轻度遮光条件下,中午时泡沙参叶片的孔开度逐渐减小和部分气关闭;裂叶沙参光合作用的非气孔限制占主要地位,泡沙参光合     

枇杷属内栽培种和部分野生种ITS序列分析

对不同栽培区的25种普通枇杷品种以及7种枇杷属野生种的ITS序列进行扩增并测序,采用邻接法和最大简约法进行系统发育树的构建并对枇杷属内不同种间的遗传关系进行了分析。结果表明:枇杷属植物ITS序列ITS1+5.8S rDNA+ITS2总长度为592 bp或594 bp,长度变化发生在ITS2。所有样本的ITS1和5.8S rDNA长度一样,都是223 bp和168 bp;而ITS2为201 bp或203 bp。5种枇杷属野生种的ITS序列长度为594 bp,包括栎叶枇杷、大渡河枇杷、南亚枇杷、南亚枇杷窄叶变种和大瑶山枇杷;其余2种枇杷属野生种(麻栗坡枇杷、小叶枇杷)和普通枇杷栽培种的ITS序列长度都为592 bp。所有样本ITS序列的GC含量为64.2%~64.5%,其中ITS1为64.1%~65.5%,ITS2为68.1%~72.6%。对所有样本的ITS序列比对产生44个可变位点,其中38个为简约信息位点,其中11个位于ITS1,5个位于5.8S rDNA,22个位于ITS2。最大的种间序列差异为7.7%,最小的种间差异发生在麻栗坡枇杷和小叶枇杷之间,仅为0.2%。普通枇杷种内的ITS序列差异很低,25种普通枇杷栽培种之间的序列差异为0~1.5%。所研究的枇杷属植物可分为3个分支。分支Ⅰ包括所有普通枇杷品种,分支Ⅱ包含5种野生枇杷种,包括栎叶枇杷、大渡河枇杷、南亚枇杷、南亚枇杷窄叶变种和大瑶山枇杷;分支Ⅲ由2个野生枇杷种(麻栗坡枇杷、小叶枇杷)组成。该研究结果表明ITS序列对枇杷种间鉴定和系统发育分析具有一定意义,但对普通枇杷栽培种间的鉴定作用不大。     

A Reevaluation of the Status of A. lobophylla Based on ITS Sequence,with Reference to the Utility of ITS Sequence in Adenophora

Adenophora Fisch. (Campanulaceae)is a medium-sized and diverse genus, which occurs mainly in eastern Asia, particularly in China. Although several taxonomic systems have been proposed, there is still much debate on the delimitation and systematic arrangement of the species within the genus, due mainly to high variation in morphology, habitats, phenotypic plasticity, and potential hybridization. A. lobophylla Hong is a rare perennial endemic to several counties of Sichuan, China. Since this species was recognized, much work has been done on its taxonomic status, including morphology, crossing experiment and allozymes. However, evidence available is not concordance so far, and its relationship with other species within the genus remains largely uncertain. In the present paper, the internal transcribed spacer(ITS) region of 18s～26s nuclear ribosomal DNA was sequenced in 10 representatives of Adenophora and two outgroup species of Campanula to assess its utility for phylogenetic reconstruction in Adenophora, in addition to reevaluation of the taxonomic status of A. lobophylla. The results indicate that there exist high sequence alignability and length conservation among ITS sequences, with informative sites being 3.9% and length ranging from 539 to 541 bp and G + C content varying from 57% to 60%. Phylogenetic　analyses using Fitch parsimony show that A. lobophylla is closely related to A. himalayana instead of A. potaninii or A. stenanthina as revealed by previous morphological work and crossing experiment. The ITS phylogeny is generally congruent with other evidence in that A. lobophylla should be removed from the A. potaninii complex or even subsect. Microdiscus. Its appropriate placement, however, should be further explored. In pairwise comparisons at unambiguously aligned sites, sequence divergence was relatively low among Adenophora species with the values ranging from 0.0 to 3.9% in spite of the fact that the species surveyed covered all of the sections and subsections of the genus. In contrast, high sequence divergence is found among Adenophora and Campanula species with the values from 17.8 % to 19.2 %. The low divergence among Adenophora species is justified considering its recent origin, and also suggests that ITS sequence would provide a promising source of nuclear phylogenetic markers to reconstruct the phylogeny within Campanulaceae on the tribe and family levels.     

Phylogeny of Hanabusaya (Campanulaceae), a Korean endemic,based on ITS sequences of nuclear ribosomal DNA

The phylogenetic position of the genus Hanabusaya was verified using the internal transcribed spacer (ITS) sequences data of 181 individuals of 176 taxa in Campanulaceae, including allied genera such as genus Campanula, Adenophora, Symphyandra, and Zeugandra. The phylogenetic analysis showed thatHanabusaya is more closely allied to Adenophora than to Symphyandra, Zeugandra, and Campanula. Accordingly, the connate anthers of Hanabusaya are considered to have evolved independently of Symphyandra or Zeugandra. However, Hanabusaya and sect. Remotiflorae of Adenophora were unresolved within a clade. Therefore, the relationship between Hanabusaya and its allies should be more accurately determined through a broader study on diverse genetic regions.     

Biosystematic studies on Adenophora potaninii Korsh.complex (Campanulaceae) V.A taxonomic treatment

Adenophora is an extremely variable genus,and its taxonomy is very controversial.Of the genus,Adenophora potaninii complex,including A.potaninii,A.bockiana,A.wawreana,A.lobophylla,A.biformifolia,A,polydentata,and A.wawreana var.lanceifolia,is a typical group with different taxonomical treatments due to high level of morphological diversity.We carried out extensive biosystematic studies based on population sampling,transplantation experiments and offspring tests,cluster analysis,and a crossing experiment.The results reveal four main findings.(i) Leaf forms of the A.potaninii complex were extremely polymorphic; the leaf form of A.potaninii and A.bockiana,and that ofA.wawreana and A.biformifolia could be found,respectively,on a single population or among the offsprings of a single plant.(ii) Cluster analysis and a crossing experiment indicated that A.bockiana and A.polydentata could not be separated from A.potaninii,nor d.biformifolia from A.wawreana.(iii) Adenophora potaninii and A.wawreana were gradational in morphology and their compatibility value was slightly reduced compared to that within each entity.(iv) Adenophora lobophylla was distinct from the other members of the complex in shape and size of corolla,relative length of style,and shape of capsule.This species was incompatible reproductively with the other members of the complex,but partly compatible with A.stenanthina,a species in another section.Therefore,we recognized only one species with two subspecies in the complex,A.potaninii subsp.potaninii and subsp,wawreana,moved A.lobophylla out of the complex,and reduced all the other names as new synonyms.     

裂叶沙参与泡沙参解剖生态学的比较研究

借助于光镜和扫描电镜,系统观察了濒危植物裂叶沙参(Adenophora lobo-phylla Hong.)的根、芦头、茎和叶的解剖特征及茎、叶、种子的表面形态特征,并将有关性状与广布种泡沙参(A.potaninii Korsh.)作了对比研究。结果表明,两种沙参在叶表皮细胞形状,垂周壁式样,角质层初级雕纹及次级雕纹;叶片气孔形状及外拱盖外缘纹饰及内缘形状,保卫细胞上角质层薄厚及蜡质纹饰;叶表毛状体形状及纹饰,茎中皮层占茎横切面的比例;茎中有无髓纤维等方面存在着明显差异。相对于泡沙参,裂叶沙参单株叶面积大,生物量高,表皮角质层薄,单位面积内被毛数量少,栅栏组织薄,气孔周围蜡质沉积颗粒少;茎的直径大,皮层较薄等。这些特征表明裂叶沙参对旱生生境适应能力弱,而泡沙参对旱生生境适应能力强。     

中国特有植物同钟花( 桔梗科) 的细胞学研究

对分布我国西南的特有属同钟花属的同钟花( Homocodon brevipes) 进行细胞学研究, 其染色体数目为68 , 核型为2n = 68 = 56m+ 10sm + 2st, 可能是四倍体。通过比较桔梗科已有的大量细胞学资料认为该类群应该与风铃草属和沙参属所代表的类群关系较近。     

中国特有植物同钟花(桔梗科)的细胞学研究

对分布我国西南的特有属同钟花属的同钟花(Homocodon brevipes)进行细胞学研究,其染色体数目为68,核型为2n=68=56m 10sm 2st,可能是四倍体。通过比较桔梗科已有的大量细胞学资料认为该类群应该与风铃草属和沙参属所代表的类群关系较近。     

Sporogenesis and Gametogenesis of Endangered Adenophora lobophylla Hong (Campanulaceae)

Reported in this paper are the microsporogenesis, megasporogenesis and the development of male and female gametophytes of the endangered species Adenophora lobophylla. The anther is four-sporangiate with its wall composed of four layers: epidermis, fibrous endothelium, middle layer and glandular tapetum of binucleate cells. The cytokinesis of microspore mother cell in meiosis is simultaneous, and tetrads are tetrahedral. Pollen grains are 2-celled at the dispersal stage. Ovule is anatropous, unitegmic and tenuinucellate with a hypodermal archesporial cell developing directly as the megaspore mother cell which undergoes meiotic division and then forms a linear tetrad. The embryo sac is of Polygonum type. The polar nuclei fuse before fertilization. An endothelium differentiates when the uninucleate embryo sac forms. Comparative studies were made with the closely related and widely distributed species A. potaninii. No differences between them were found. No obstacle to sexual reproduction in this endangered species was observed. As a conclusion, no endangering factors were found to influence the sporogenesis and gametogenesis of A. lobophylla. According to our observision and the references on embryological studies of the Campanulaceae(s. l. ), there is no evident differentiation in sporogenesis and gametogenesis in this family.