菊花起源的RAPD分析

利用ＲＡＰＤ分析技术,选取２２个２０个碱基长度的随机引物,对７种野生菊花、１４种栽培菊花和５个种间杂种进行了随机扩增。通过实验建立了ＰＣＲ随机扩增实验体系。在观察到的２２４个扩增条带中,３４条（１５％）表现多态性。利用ＵＰＧＭＡ法对扩增数据进行分析,结果表明：在７种野生种中,Ｄｅｎｄｒａｎｔｈｅｍａｉｎｄｉｃｕｍ、Ｄ．ｖｅｓｔｉｔｕｍ和Ｄ．ｎａｎｋｉｎｇｅｎｓｅ与栽培菊花亲缘关系最近,而Ｄ．ｚａｗａｄｓｋｉ与栽培菊花亲缘关系最远。前３种野菊与栽培菊花间的遗传距离小于０．４０,而Ｄ．ｚａｗａｄｓｋｉ与栽培菊花间的遗传距离大于０．５０。根据上述数据及以往研究结果,使用ＲＡＰＤ数据对菊花起源问题进行了探讨,提出栽培菊花主要起源于Ｄ．ｉｎｄｉｃｕｍ、Ｄ．ｖｅｓｔｉｔｕｍ和Ｄ．ｎａｎｋｉｎｇｅｎｓｅ．     

Origins of cultivars of Chrysanthemum—Evidence from the chloroplast genome and nuclear LFY gene

The origins of cultivated chrysanthemums have attracted considerable attention, but they remain poorly known. Here, we reconstructed the phylogeny of representative well‐known cultivars and wild species of the genus Chrysanthemum using chloroplast genomes and the nuclear LEAFY gene. Our results suggest that geographic and ecological factors may determine the opportunities for wild species to be involved in the origin of the cultivars. The wild species C. indicum, C. zawadskii, C. dichrum, C. nankingense, C. argyrophyllum, and C. vestitum were likely directly or indirectly involved as paternal species of most of the chrysanthemum cultivars examined in this study. Yet, the maternal species is supported to be a lineage of an extinct wild Chrysanthemum species and its subsequent cultivars, as all accessions of chrysanthemum cultivars sampled formed a strongly supported clade, distinct from all other species of Chrysanthemum in the plastome tree. Thus, the cultivated chrysanthemums originated from multiple hybridizations involving several paternal species rather than only two or a few wild species, with an extinct species and its subsequent cultivars serving as the maternal parents. This finding is consistent with Chrysanthemum having high rates of hybridization and gene flow, which has been demonstrated within previous studies; nevertheless, it is important to unravel the role of an extinct wild Chrysanthemum species as the ultimate maternal parent species for all the chrysanthemum cultivars. Our results also suggest that C. vestitum from Tianzhu and Funiu Mountains in Anhui and Henan Provinces of China represent two distinct cryptic species.     

安徽野生菊属植物叶下表面特征的研究

利用光学显微镜对5种安徽野生菊属(Chrysanthemum)植物(野菊、甘菊、菊花脑、毛华菊和紫花野菊)的叶下表面特征进行研究,并探索与植物分布、生态的相关性.结果显示,这些类群非腺毛、腺毛的形态、大小和密度均有差异;气孔的大小和密度、表皮细胞的大小和角质纹理等方面也存在差异.叶片表面的特征与植物生长环境有相关性.依据这些特征讨论了菊花脑的分类并尝试性地编制了这些类群的检索表.     

Phylogenetic Relationship of Dendranthema (DC.) Des Moul. Revealed by Fluorescent In Situ Hybridization

Phylogenetic relationships of the different species in the genus Dendranthema (DC.) Des Moul. were estimated based on chromosome fluorescent in situ hybridization (FISH) with 18S-26S rDNA of Arabidopsis and genomic DNA of Dendranthema as probes. The results revealed that there was no positive correlation between the number of nuclear organization region (NOR) loci and the ploidy of Dendranthema.The exact cytogenetic information of NORs about 14 operational taxonomic units (OTUs) indicated that D.vestitum (Hemsl.) Ling et Shih was closer to the cultivars than other putative species, whereas D. zawadskii (Herb.) Tzvel. was the most distinct. The ambiguously distributed signals of genomic in situ hybridization (GISH) with genomic DNA of lower ploidy species as probes suggested that different genomes among Dendranthema were mixed. The result also indicated the limitation of GISH in studies on the phylogenetic relationships of the different species in this genus Dendranthema and on the origin of cultivated chrysanthemums. Based on these results and previous research, the origin of Chinese cultivated chrysanthemum is discussed.     

Phylogenetic Relationship of Dendranthema （DC.） Des Moul. Revealed by Fluorescent In Situ Hybridization

Phylogenetic relationships of the different species in the genus Dendranthema (DC.) Des Moul. were estimated based on chromosome fluorescent in situ hybridization (FISH) with 18S-26S rDNA of Arabidopsis and genomic DNA of Dendranthema as probes. The results revealed that there was no positive correlation between the number of nuclear organization region (NOR) loci and the ploidy of Dendranthema.The exact cytogenetic information of NORs about 14 operational taxonomic units (OTUs) indicated that D.vestitum (Hemsl.) Ling et Shih was closer to the cultivars than other putative species, whereas D. zawadskii (Herb.) Tzvel. was the most distinct. The ambiguously distributed signals of genomic in situ hybridization (GISH) with genomic DNA of lower ploidy species as probes suggested that different genomes among Dendranthema were mixed. The result also indicated the limitation of GISH in studies on the phylogenetic relationships of the different species in this genus Dendranthema and on the origin of cultivated chrysanthemums. Based on these results and previous research, the origin of Chinese cultivated chrysanthemum is discussed.     

部分菊属植物及其种间杂种减数分裂异常现象观察

对不同倍数性菊属植物及其部分种间杂种的减数分裂异常现象进行观察统计,并分析其形成机制以及在菊属系统演化中的作用。结果表明,菊属减数分裂异常现象包括分裂不同步、二价体提前解离、二价体互锁、染色体桥、落后染色体等。减数分裂不同步现象普遍存在于菊属植物减数分裂过程。二倍体的菊花脑、甘菊、异色菊的部分二价体在终变期提前解离为单价体。菊花脑及其部分杂种中观察到了互锁二价体。四倍体菊花脑、南京野菊、‘黄英’、‘滁菊’在AI和AII都出现了染色体桥,毛华菊有1.5%的PMC在AI出现染色体桥。四倍体菊花脑AI、AII期出现落后染色体的频率分别为10.6%和7.3%;毛华菊AI期出现落后染色体的频率为4.4%;栽培菊‘黄英’和‘滁菊’在AI、AII期出现落后染色体的频率高于毛华菊。杂种出现染色体桥及落后染色体的频率普遍高于亲本。倒位以及由其引起的各种染色体结构变异可能在菊属系统演化过程中起着重要作用。     

用RAPD和ISSR检测的橡胶树野生种质和栽培品种的遗传多样性

用19个RAPD引物和12个ISSR引物对14份野牛橡胶树种质和我国的37份栽培品种进行了遗传多样性分析。RAPD引物共产生132条带,多态性带占88．6％,相似系数变化范围在0．432—0．947。ISSR引物其产生101条带,多态性带占87．1％,相似系数为0．505—0．941。平均基因杂合度分析表明野生种质比栽培品种具有较高的遗传多样性。根据UPGMA法对51份材料进行聚类分析,结果表明,ISSR分析中所有材料可分为2类：第一类为野生种质,第二类为栽培品种：而RAPD分析中野牛种质和栽培品种不能被分为明显的两人类。虽然ISSR和RAPD的聚类分析结果存在差异,但对两种方法进行的相关分析表明,他们之间仍存在极显著相关性,相关系数为0．574。品种PR107、热研217等一些栽培品种可以通过特异带在51份供试材料中被区分开。这些结果可以对橡胶树的育种上作起到一定的指导作用,同时RAPD和ISSR技术也是进行橡胶树品种鉴定和遗传多样性研究的有效手段。     

Identification of wild and cultivated Hordeum species using two-primer RAPD fragments

Random amplified polymorphic DNAs (RAPD) analysis has been adapted to assess the degree of RAPD polymorphism within the genus Hordeum to determine if this approach can distinguish wild and cultivated species. Nineteen wild and seven cultivated accessions were evaluated using 4 random 10-mer primers. The potential of the RAPD assay was further increased by combining two primers in a single polymerase chain reaction (PCR). RAPD fragments generated by two pairs of arbitrary 10-mer primers discriminated six wild species and one cultivated species by banding profiles. The size of the amplified DNA fragments ranged from 150 to 2300 base pairs. 33 %percent of the fragments were common to both wild and cultivated species; 67% were specific to either wild or cultivated species. The average difference in fragments was less within the species than among the species. By comparing RAPD fingerprints of wild and cultivated barley, markers were identified among the set of amplified DNA fragments which could be used to distinguish wild and cultivated Hordeum species. This revised version was published online in July 2006 with corrections to the Cover Date.     

海南主要普通野生稻的调查及利用RAPD技术进行鉴别分析

通过对海南现存的主要普通野生稻资源进行考察,报道了海南主要普通野生稻的最新生存状况;并用RAPD技术对其进行分析。从供试材料中筛选出具有多态性的RAPD引物13条,共扩增出155条带,其中多态性条带120条,多态性条带比率(PPB)值为77.42%。UPGMA聚类结果表明:海南主要普通野生稻之间遗传关系较复杂、差异较明显,并将海南主要普通野生稻资源划分为5个小类群,可以很好地将海南本地野生稻与收集保存的野生稻鉴别开来,为海南普通野生稻资源的合理利用与有效保护提供了重要依据。     

野生稻和栽培稻的随机多态DNA(RAPD)分析

应用 RAPD方法对药用野生稻、普通野生稻、粳稻和籼稻进行基因组多态性分析。 1 2个随机引物共扩增出 1 3 2条 RAPD带 ,片段大小在 3 0 0～ 3 5 0 0 bp之间 ,其中有 1 0 6条表现出多态性 ,占总扩增片段的86.4%。根据遗传距离分析 ,用 UPGMA法构建了聚类树状图 ,结果表明 ,普通野生稻的遗传特性比药用野生稻更接近于栽培稻。     

AFLP analysis of genetic polymorphism and evolutionary relationships among cultivated and wild Nicotiana species.

Amplified fragment length polymorphism (AFLP) analysis was used to determine the degree of intra- and inter-specific genetic variation in the genus Nicotiana. Forty-six lines of cultivated tobacco (Nicotiana tabacum L.) and seven wild Nicotiana species, including N. sylvestris, N. tomentosiformis, N. otophora, N. glutinosa, N. suaveolens, N. rustica, and N. longiflora, were analyzed, using at least eight different oligonucleotide primer combinations capable of detecting a minimum of 50 polymorphic bands per primer pair. The amount of genetic polymorphism present among cultivated tobacco lines (N. tabacum) was limited, as evidenced by the high degree of similarity in the AFLP profiles of cultivars collected worldwide. Six major clusters were found within cultivated tobacco that were primarily based upon geographic origin and manufacturing quality traits. A greater amount of genetic polymorphism exists among wild species of Nicotiana than among cultivated forms. Pairwise comparisons of the AFLP profiles of wild and cultivated Nicotiana species show that polymorphic bands present in N. tabacum can be found in at least one of three proposed wild progenitor species (i.e., N. sylvestris, N. tomentosiformis, and N. otophora). This observation provides additional support for these species contributing to the origin of N. tabacum.     

橡胶树种质资源遗传多样性研究——Ⅰ．速生种质遗传多样性RAPD分析

应用RAPD技术对8份野生种质和12份栽培种质进行遗传多样性分析,筛选到18个具有多态性扩增的引物．共扩增出128条带。据Nei-Li相似系数将20份材料分别聚为野生种质和栽培种质两大类。5个野生种质聚为野生种质类群,12个栽培种质和3个野生种质聚为栽培种质类群。研究结果表明,RAPD技术用于橡胶树种质资源研究,能够为野生种质优良特性导入栽培种质提供分子水平的参考依据。     

Physical mapping of rDNA in Dendranthema nankingense and its close related species by fluorescent in situ hybridization

Distribution of 18S-26S rDNA on the chromosomes of Dendranthema nankingense and its two close-related species D. nankingense and D. lavendulifolium was studied by fluorescent in situ hybridization with a JHD 2-15A DNA clone. Six rDNA loci were found in D. nankingense, eight in D. lavendulifolium, and twelve in D. indicum. The rDNA loci number and pattern were applied to analyze the phylogenetic relationship of the three closely related species.     

野生稻基因组随机扩增多态性DNA(RAPD)分析

用18个随机引物对2份栽培稻、12份包含有六个基因组型的野生稻DNA进行了扩增,共获得147个多态性DNA片断,把这些多态性DNA片断作为遗传位点用UPGMA法计算出各材料间的遗传相似性系数,并作了聚类分析．主要结果如下：1普通野生稻同栽培稻的亲缘关系很近,其中江永普通野生稻更接近于粳稻．2．CCDD组的Oryzalatifolia和EE组的O．australiensis遗传多态性相似。3．B、C、D、E组的遗传多态性相似,组成一个复合体,此复合体与A组的遗传多态性也相似,而F组则相距较远．4．O．mcyeriana和Rhynchofyzasabulata尚未确定组型,RAPD测定结果表明,前者与其它组型的种亲缘关系较远,后者则与AC复合体的种较近．     

Efficacy of random primer-pair arrays in plant genome analysis: a case study of Cucumis (Cucurbitaceae) for identification of wild and cultivated species

The efficacy of random primer-pair arrays compared to conventional RAPD method with a single decamer primer was evaluated using DNA from two species of Cucumis. The banding patterns of amplicons revealed enhanced utility of primer-pair arrays over conventional RAPDs, producing more bands and a higher degree of polymorphism, both at intra- and inter-specific levels. Amplification produced by both methods clearly distinguished a wild from a cultivated species of the genus Cucumis. The main advantage of the primer-pair RAPD over single-primer-based RAPD is the increase in the number of reactions and amplification products in the form of novel/unique bands with a limited number of primers. It also enables the generation of reliable amplicons with a large number of polymorphic bands, which can be linked to gene-governing traits, allowing sequence-characterized partial genome analysis.     

羊肚菌栽培菌株遗传多样性分析及种特异性RAPD-SCAR标记开发

近年来中国的羊肚菌Morchella spp.栽培技术取得了长足进步,但基础研究薄弱影响其稳产和高产,国内外尚无羊肚菌栽培菌株种质资源遗传多样性的研究报道。本文对来自全国12省份的36个羊肚菌栽培菌株进行了ITS系统发育分析,并采用RAPD进行了遗传多样性评价。结果表明,结合有效的参考菌株序列,通过ITS序列分析可以将供试菌株进行区分和鉴定,在36个菌株中,26个菌株属于梯棱羊肚菌Morchella importuna,其他10个菌株属于六妹羊肚菌M. sextelata;将自40条RAPD引物中筛选出的14条用于供试菌株遗传多样性分析,共扩增出124条多态性条带;UPGMA聚类可将供试菌株分为两大类群,分别对应于ITS系统发育分析中的梯棱羊肚菌和六妹羊肚菌两个物种,梯棱羊肚菌种内菌株多态性高于六妹羊肚菌。OPA17引物和OPA18引物分别在AA02和AA15菌株中扩增出具有唯一性的特征条带,对两个特征条带进行回收测序后,设计出两个特异性SCAR的引物,它们能有效地从36个供试菌株群体中将菌株AA02和AA15鉴别出来。本文首次全面系统地采用ITS分析鉴别了我国羊肚菌栽培菌株的种性,采用RAPD分子标记系统地评价了羊肚菌栽培菌株的遗传多样性,并验证了RAPD分子标记转化为菌株特征性SCAR标记的可行性。     

两种菊属植物对渐进与非渐进盐胁迫的响应

以两种菊属野生植物'菊花脑'(Dendranthema nankingense)和'乙立寒菊'(D.indicum var.maruyamanum)幼苗为材料,通过Hogland营养液水培试验研究了120和180 mmol·L NaCl的渐进和非渐进胁迫处理对幼苗叶片受害程度及植株生物量、根系活力、叶片叶绿素含量、相对电导率和丙二醛含量的影响,以探讨它们对盐冲击的生理响应规律.结果显示:(1)在两种浓度盐胁迫下,两种菊属植物的整株鲜重、根系活力、叶片叶绿素含量大多显著下降,而同期的叶片丙二醛(MDA)含量、相对电导率显著增加;渐进盐胁迫下材料受害程度均轻于非渐进胁迫;盐胁迫浓度越大,各指标变化幅度越大,且两种胁迫方式下各指标的差异亦越大;(2)'菊花脑'对盐胁迫敏感,而'乙立寒菊'对盐胁迫的耐性较强;相同盐胁迫浓度下,非渐进胁迫与渐进胁迫间的差异在盐敏感材料上表现较明显.研究表明,两种菊属植物均存在盐冲击现象,并以耐盐性弱的材料表现更加突出,且高盐胁迫下植物所受的冲击伤害更明显.     

用RAPD技术探讨中国枣的种下划分

利用随机扩增多态性DNA(RAPD)技术对14个枣Ziziphus jujuba 品种和1个野生种——泰山酸枣Z．spinosus的遗传变异进行了研究。从120个10-碱基随机引物中筛选出37个多态性引物用于正式扩增,共扩增出429条DNA带,其中多态性带214条,占49．88％。根据DNA扩增结果计算了品种及类型间遗传距离,并用UPGMA构建了聚类树状图。分析结果表明：龙爪枣 Z．jujuba var．tortuosa、葫芦枣 Z. jujuba var．lageniformis、无核枣 Z.jujuba var．anucleatus 等几个变种内的遗传距离大于变种间遗传距离,认为枣的变种划分是不自然的,宜并入其原变种;枣种下不宜设变种,对枣种下的众多品种,应根据品种间的遗传关系,直接划分品种群。     

应用RAPD分子标记技术探讨3种石斛属植物的种间关系

采用RAPD分子标记技术,分析了金钗石斛、铁皮石斛和齿瓣石斛三种石斛属植物的种间关系。10个引物产生的113条DNA扩增片段中,106条（93.81%）具有多态性,利用113个RAPD标记,计算遗传距离,利用非加权组平均法建立聚类图。结果表明,RAPD标记技术较好地从分子水平揭示金钗石斛、铁皮石斛和齿瓣石斛三种石斛属植物的遗传背景、亲缘关系,并为后期在DNA水平上对药用石斛的开发利用提供资料。     

不同金银花种源间遗传关系的RAPD分析

利用RAPD分析技术对金银花(Lonicera japonicaThunb.)6个种源的遗传多样性及遗传关系进行了研究。结果表明,23个引物共扩增出105条DNA片段,其中,多态性片段83条,占扩增总数的79.05%。聚类分析结果表明,金银花的6个种源可聚为2大类,产自山东平邑、湖南隆回和江苏南京的野生金银花种源遗传距离较近,聚为一类;来自河南封丘的2个野生种源和来自河南密县的1个栽培种源聚为一类。不同金银花种源间的遗传关系与地理分布有一定的相关性。