基于多基因序列和形态性状的牡丹组种间关系

牡丹被认为是中国的国花,具有很高的医学、观赏和经济价值.野生牡丹被认为是栽培牡丹的野生祖先,因此弄清牡丹组的种间亲缘关系具有重要的理论和实践意义.由于受到信息量的限制,根据单基凼数据或形态数据往往无法对牡丹组的种间关系得到明确的结果.本研究用12份样品代表野生牡丹组(Paeonia section Moutan DC.,Paeoniaceae)8个种,利用包括核基因(Adh1A、Adh2和GPAT)和叶绿体基因(trnS-trnG和rps16-trnQ)的DNA序列以及形态性状的多套数据来探讨野牛牡丹的种间关系.合并分析得到具高支持率的牡丹组物种间的系统发育关系.结果表明,芍药属牡丹组8个野生种分为两个亚组,即肉质花盘亚组subseet.Delavayanae和革质花盘亚组subsect.Vaginatae.肉质花盘亚组包括滇牡丹P delavayi和大花黄牡丹P.ludlowii;革质花盘亚组包括其余6个种.革质花盘亚组中,四川牡丹P.decomposita ssp.decomposita和紫斑牡丹P. rockii ssp. rockii关系密切;卵叶牡丹P.qiui和矮牡丹P. jishanenMs关系密切;银屏牡丹P. suffruticosa ssp.yinpingmudan与风丹P. ostii关系 密切,并且后两个分支为姊妹群.     

基于ITS和matK基因对牡丹组序列分析及其亲缘关系的研究

以芍药属牡丹组全部9个野生种、5个紫斑牡丹栽培品种及3个中原牡丹品种为试材,进行核糖体内转录间隔区（ITS）和叶绿体成熟酶K （matK）基因片段测序分析,探讨ITS和matK序列为牡丹组所有野生种种间关系提供分子证据。从GeneBank中选取了1个牡丹及3个外类群芍药、川赤芍和草芍药的ITS及matK序列。对试验样品进行DNA提取、PCR扩增并双向测序得到44条序列,人工校正后将所得44条序列进行比对;计算碱基组成频率、变异位点、简约信息位点数、转换/颠换比率、种内及种间遗传距离,以邻接法进行系统发育分析。结果表明,牡丹组所有个体ITS序列长度在750~800 bp,含有86个多态位点,74个简约信息位点,转换/颠换比率（R）为1.2;而matK序列含有20个简约信息位点,转换/颠换比率（R）为1.7。ITS序列分析将牡丹组野生种分为两大枝,稷山牡丹、紫斑牡丹、卵叶牡丹和杨山牡丹聚为一枝,狭叶牡丹、滇牡丹、黄牡丹和大花黄牡丹聚为另一枝,这两枝分别与革质花盘亚组和肉质花盘亚组相对应,而四川牡丹位于革质花盘亚组最底端,支持前人研究将四川牡丹归为革质花盘亚组。matK序列分析的牡丹组野生种间遗传距离结果不理想,未能清晰的表明野生种之间的亲缘关系。由此说明,ITS序列更适合牡丹组野生种间亲缘关系的研究分析。     

革质花盘亚组野生牡丹资源的调查及保护利用建议

革质花盘亚组野生种参与了栽培牡丹的起源,在生物多样性保护和牡丹品种改良中具有重要价值。为了科学合理地保护和开发利用我国特有的野生牡丹资源,本研究通过野外实地调查,结合文献记录,在全国尺度上对革质花盘亚组野生牡丹资源的生物学特性、地理分布特点进行了系统分析。结果表明:革质花盘亚组共有5个野生种和1个杂交种,分布在秦巴山区、陕甘黄土高原和川西北黄土高原地区,其中陕西省野生牡丹资源最为丰富,并在陕西省商南县和旬阳县发现了卵叶牡丹新的原生地分布点;同时对革质花盘亚组的种群特征进行了分析,并针对革质花盘亚组种质资源的药用、观赏和油用潜在利用价值和濒危现状提出了开发利用建议和保护对策。     

野生牡丹的生存状况和保护

芍药属(Paeonia L.)的木本植物属于牡丹组(Paeonia sect.Moutan DC.),共有9个野生物种和1个包含上千品种的栽培种。所有野生牡丹都为中国特有,是非同寻常的资源植物。1992年出版的《中国植物红皮书》收录了388个保护类群,野生牡丹就占了4个。本文回顾了野生牡丹保护的历史、调查和研究的经过、研究结果以及存在的问题,着重介绍了野生牡丹的生物学背景、分布和生境,并就其濒危等级和保护措施提出了建议。根据我们的调查和零星的资料,凤丹(P.ostii)和中原牡丹(P.cathayana)都只剩单株,亟待挽救;紫斑牡丹(P.rockii)、卵叶牡丹(P.qiui)、四川牡丹(P.decomposita)和圆裂牡丹(P.rotundiloba)处于濒危状态;大花黄牡丹(P.ludlowii)和矮牡丹(P.jishanensis)应属于易危等级,只有滇牡丹(P.delavayi)尚不在濒危之列。总结以往保护的进程,我们强调,如果没有科学的物种划分,保护单元不明确,生态和生物学信息不够充实,就难以制定科学的保护策略和可行的保护措施。因此,呼吁有关部门高度重视这一问题,开展深入的野生牡丹现状的调查,制定切实可行的保护策略,并实行有力的保护措施。     

基于系统发育分析的DNA条形码技术在澄清芍药属牡丹组物种问题中的应用

清晰的物种概念是材料正确鉴定的基础,是DNA条形码技术应用的前提.本文通过芍药属牡丹组植物DNA条形码数据的系统发育分析,揭示牡丹组植物的进化谱系及其与分类上“物种”的关系.在此基础上分析DNA条形码技术在牡丹组植物中应用的可能性.同时,以芍药科芍药属牡丹组植物为例,讨论DNA条形码技术在应用中存在的问题与解决方案.研究材料包括牡丹组植物目前已知的几乎所有的变异类型（种或种下分类群）共40份.DNA序列来自叶绿体基因组ndhF,rps16-trnQ,trnL-F和trnS-G4个基因,共有（含插入/缺失）5040个位点,包含96个变异位点,其中信息位点69个;核基因组GPAT基因2093~2197bp,变异位点（含插入/缺失）总数279个,其中信息位点148个.叶绿体基因组与核基因组所揭示的包括四川牡丹、矮牡丹、卵叶牡丹和紫斑牡丹在内的进化线与根据形态所建立的物种限定不吻合：（ⅰ）叶绿体基因分化明显但核基因没有明显分化——四川牡丹和紫斑牡丹的各居群系统;（ⅱ）核基因分化显著而叶绿体基因分化很小——矮牡丹和卵叶牡丹.因为这些居群系统之间存在一定程度的地理隔离,但不存在生殖隔离,出现这种两个基因组数据背离的现象可能是由于不同居群系统在进化历史中捕获了其他居群系统的叶绿体基因组.这些基因作为牡丹组植物DNA条形码的适合性分析显示,叶绿体基因在种间或居群系统之间的变异是种或居群系统内变异的4.82倍,GPAT基因在种间或居群系统之间的变异是种或居群系统内变异的2.21倍,说明这些基因可作为牡丹组植物的DNA条形码.种间或居群系统之间完全分化位点的统计结果表明,这些种或居群系统之间存在相互区别所必需的位点.通过牡丹组植物DNA条形码分析,认为：（ⅰ）物种概念对DNA条形码技术能否成功应用有十分重要的影响,拟应用DNA条形码的类?     

中国野生牡丹研究 二：芍药属牡丹组新分类群

本文发表一个新亚种和一个新等级。新亚种林氏牡丹和紫斑牡丹同属一个种系,花瓣基部均具紫斑,花丝,花盘及柱头均为白色;但林氏牡丹的小叶多为披针形或窄卵形,全缘,而紫斑牡丹的小叶多为卵形或卵圆形,常１－３深裂。     

芍药属牡丹组基于形态学证据的系统发育关系分析

对芍药属牡丹组Paeonia L.sect.Moutan DC.（全部野生种）40个居群进行了基于形态学证据的系统学分析,试图建立组内种间的系统发育关系。利用PAUP （4.0）计算机程序分别构建了建立在25个形态学性状基础上的所有研究类群的距离树（UPGMA、NJ）和最大简约树（MP）。所得树的拓扑结构基本一致,差异只发生在距离树和简约树之间,在由形态和细胞学关系都很近的5个种（牡丹P.suffruticosa、矮牡丹P.jishanensis、卵叶牡丹P.qiui、紫斑牡丹P.rockii和凤丹P.o     

光照强度对紫斑牡丹生理及生长特性的影响

紫斑牡丹(Paeonia suffruticosa var. papaveracea)是集观赏、油用及药用价值为一体的经济作物,与牡丹(Paeonia suffruticosa)相比具有较强的耐寒、耐旱性,对光照条件的要求较为苛刻。为了解紫斑牡丹对光照的需求及适应规律,本研究以3年生紫斑牡丹幼苗为材料,模拟东北地区林下光强,研究了紫斑牡丹在4种光照强度(全光照、透光率55%、25%和15%)处理下的生理及生长特性。结果表明:与全光照相比,遮阴条件下紫斑牡丹通过提高叶片色素含量、表观量子效率(AQE)、PSII最大光化学效率(Fv/Fm)及光补偿点(LCP)来提高对光能的吸收利用效率,以适应弱光环境,其中25%透光率处理的紫斑牡丹具有最高的光合性能和净光合速率(Pn),为全光照的1.58倍;在全光照处理下,紫斑牡丹叶片受到强光抑制,丙二醛(MDA)和脯氨酸(Pro)含量显著增加(P0.05),光合作用系统受到严重损害,光合速率较低。遮阴处理时紫斑牡丹通过增大叶片比叶面积(SLA)、降低根冠比(R/S)并促进植株株高生长来获取更多的光照,其中,25%透光率处理的紫斑牡丹株高生长量显著高于全光照和15%透光率处理,分别为二者的4.57和3.87倍。本研究表明,25%透光率是最有利于紫斑牡丹生长的理想光照条件,该结果为紫斑牡丹在东北林下的引种栽培提供了科学依据。     

‘挽春’(Paeonia rockii‘Wan Chun’)——中国西北紫斑牡丹品种群中的一个优良品种

紫斑牡丹品种群是中国的主要牡丹品种群之一。文中描述了一个优良紫斑牡丹品种‘挽春’(Paeonia rockii‘Wan Chun’)的特征、习性和表现,该品种在紫斑牡丹品种群中具有代表性。紫斑牡丹品种可用于园艺观赏、经济林营造、荒山绿化以及荒漠治理。根据多年的调查和相关文献,按照紫斑牡丹品种的生长适应程度和引种栽培难易程度,将中国有可能引种栽植紫斑牡丹品种的地区划分为4类。介绍了各类地区在引种栽培方面需要注意的关键技术措施,分析了紫斑牡丹品种向全国各地推广过程中存在的问题,并提出了建议。     

'挽春'(Paeonia rockii 'Wan Chun')——中国西北紫斑牡丹品种群中的一个优良品种

紫斑牡丹品种群是中国的主要牡丹品种群之一。文中描述了一个优良紫斑牡丹品种‘挽春’(Paeoniarockii‘W an Chun’)的特征、习性和表现,该品种在紫斑牡丹品种群中具有代表性。紫斑牡丹品种可用于园艺观赏、经济林营造、荒山绿化以及荒漠治理。根据多年的调查和相关文献,按照紫斑牡丹品种的生长适应程度和引种栽培难易程度,将中国有可能引种栽植紫斑牡丹品种的地区划分为4类。介绍了各类地区在引种栽培方面需要注意的关键技术措施,分析了紫斑牡丹品种向全国各地推广过程中存在的问题,并提出了建议。     

Taxonomical history and revision of Paeonia sect. Moutan (Paeoniaceae)

The taxonomical history of the woody group(peonies) in the genus Paeonia is reviewed in the present paper. The group is endemic to China, but Paeonia suffruticosa is commonly cultivated throughout the north temperate region and has long been known as “the King of Flowers” in China. However, the taxonomy of the group had been neglected before the 1990' s. Since 1990, a number of new species and subspecies have been published. With the support from the National Geographic Society, the senior author of the present paper and his coworkers have made expeditions to all parts of the distribution area of the group, visited all the type localities and sampled a total of 64 populations. Based on the character analysis and examination of the type specimens or photos, each taxon is reviewed with its name checked nomenclaturally, and finally the taxonomy of the whole group is revised. As a result, eight species, three of which each contains two subspecies, are recognized. They are Paeonia suffruticosa Andrews subsp. suffruticosa and subsp. yinpingmudan D. Y. Hong, K. Y. Pan et Z. W. Xie, P. jishanensis T. Hong et W. Z. Zhao, P. qiui Y. L. Pei et D. Y. Hong, P. ostii T. Hong et J. X. Zhang, P. rockii (S. G. Haw et L. A. Lauener )T. Hong et J. J. Li subsp. rockii and subsp. taibaishanica D. Y. Hong, P. decomposita Hand.-Mazz. Subsp. decomposita and subsp. rotundiloba D. Y. Hong, P. delavayi Franeh. and P. ludlowii (Stern et Taylor)D. Y. Hong. P. baokangensis Z. L. Dai et T. Hong and P. yananensis T. Hong et M. R. Li are considered as hybrids between P. rockii and P. qiui and between P. rockii and P. jishanensis respectively. In addition, the reduction of a number of names recently published to synonyms is explained.     

Phylogenetic analysis of Paeonia sect. Moutan (Paeoniaceae) based on multiple DNA fragments and morphological data

Tree peony, being crowned the title “King of Flowers” in China, is of great medicinal, ornamental, and economic values. In the present study, the phylogeny of the wild tree peony species (section Moutan, Paeonia, Paeoniaceae), represented by twelve accessions collected from all eight species in the section, was investigated based on the DNA sequence in five DNA fragments from both nuclear (Adh1A, Adh2 and GPAT) and chloroplast (trnS-trnG and rps16-trnQ) genomes, as well as morphological characters. Both maximum parsimony (MP) and Bayesian inference of phylogeny (BI) trees were reconstructed based on the combined data of the DNA sequences and morphological data, respectively. The MP and BI trees have the similar topology, and the sect. Moutan clearly branched into two clades. One clade consists of two species, P. delavayi and P. ludlowii, corresponding to the subsect. Delavayanae, and another clade is composed of other six species. Within the second clade, the six species can be divided into three subclades consisting of P. rockii and P. decomposita, P. jishanensis and P. qiui, P. suffruticosa and P. ostii, respectively. Among the three subclades, P. jishanensis/P. qiui is most closely related to P. suffruticosa/P. ostii. These results provide up to date the clearest picture of the phylogeny of wild tree peony species in the sect. Moutan.     

紫斑牡丹及其一新亚种

本文回顾了有关紫斑牡丹的调查和分类历史。它曾被混同于 P．suffruticosa,P．papaveracea 及P．suffruticosa var．papaveracea。它以叶2至3回羽状复叶,小叶17～33,花瓣白色,基部有大紫斑, 花丝黄色,花盘和柱头淡黄色区别于近缘种。种下分化为两个异域的亚种;秦岭北坡的紫斑牡丹小叶全 部或大部分分裂,是一个新亚种,P．rockii subsp．taibaishanica,而P.rockii subsp．linyanshanii T.Hong et G．L.ostii则是P．rockii subsp．rockii的多余名。     

Paeonia rockii and Its One New Subspecies from Mt. Taibai,Shaanxi of China

The history of exploration and classification of Paeonia rockii is reviewed in the present paper. Two allopatric subspecies in this species are recognized. The new subspecies, subsp, taibaishanica Hong, found on northern slope of the Qingling Range has leaflets ovate or rounded, totally or mostly lobed. P. rockii subsp, linyanshanii T. Hong et G. L. Ostiis found to be a superfluous name of P. rockii subsp, rockii.     

A revision of the Paeonia suffruticosa complex (Paeoniaceae)

The taxonomical concept of the Paeonia suffruticosa complex i.e. Sect. Moutan Subsect. Vaginatae, has changed greatly since 1990. Six species and four subspecies have been described as new and two subspecies raised to specific level. Five species and two subspecies are recognized in the present revision, viz. P. suffruticosa subsp. suffruticosa and subsp. yinpingmudan, P. jishanensis, P. qiui, P. ostii, P. rockii subsp. rockii and subsp. taibaishanica. P. yananensis, P. ridleyi, P. spontanea, P. moutan subsp. atava, P. suffruticosa subsp. atava, P. rockii subsp. linyanshanii and P. ostii var. lishizhenii are treated as synonyms. P. papaveracea and P. baokangensis are proposed to be interspecific hybrids. A key to the recognized species and subspecies is provided. Biological features of the species are described and their distributions are mapped. The relationships between species are inferred and the origins of commonly cultivated tree peonies ( P. suffruticosa and P. ostii ) are discussed.     

Preliminary Report of RAPD Analysis in Paeonia suffruticosa Subsp. spontanea and Paeonia rockii

Ten different oligonucleotide primers of arbitrary sequence were used in Capillary and Air Thermocycler to amplify genomic total DNA of Paeonia suffruticosa subsp. spontanea and P. rockii isolated from several local populations in Shanxi, Shaanxi and Gansu provinces. Under the strictly standardized amplification condition for all the primers, these primers yielded clear and reproducible bands corresponding to amplified products and separable by agarose gel electrophoresis. Among a total of 71 bands amplified, 16(22. 5%) were polymorphic in a single individual of P. suffruticosa subsp. spontanea, while among a total of 76 bands 21(27.6%)were polymophic in a single individual of P. rockii. On an average,the pairwise marker difference between band profiles of conspecific individuals (different populations) was 7.9 for P. suffruticosa subsp. spontanea and 8.7 for P. rockii respectively. The average marker difference between P. suffruticosa subsp. spontanea and P. rockii was 10.3. Obviously, greater number of plants and primers will be required to detect satisfactorily level of genetic diversity. These preliminary results showed that intraspecific genetic diversity was low for the two endangered species. RAPD as a molecular marker was useful and feasible for detecting the genetic variation within species of wild moutans. And it was also potential for studying evolution and relationships between species.     

矮牡丹与紫斑牡丹RAPD分析初报

用10个任意序列的寡核苷酸片段作为引物,将采自陕西、山西、甘肃等地矮牡丹与紫斑牡丹基因组DNA,在毛细管气浴式PCR热循环仪上随机扩增。在对所有引物扩增条件严格标准化的条件下,这些引物可产生清楚的、可重复的与扩增产物相应的琼脂糖凝胶电泳区带。这10个有效扩增的引物在矮牡丹平均每一个体中扩增出71条带,其中多态的带16条(22．5％),在紫斑牡丹平均每一个体中扩增出76条带,其中多态的带2l条(27．6％)。对每两个个体,每条多态带进行成对比较,累加后求分子标记差异的平均值。在矮牡丹3个居群间的平均差异是7．9,在紫斑牡丹4个居群间的平均差异是8．7,在矮牡丹与紫斑牡丹2个种间的平均差异是10.3。显然,若用来扩增的植物个体数目和引物数目增加,将会得到更满意的结果。初步结果表明濒危植物矮牡丹与紫斑牡丹种内低水平的DNA多态性。RAPD技术用于检测野生牡丹居群内与居群间的遗传变异是有用的与可行的;用于研究种间的进化和亲缘关系也有潜力。     

Systematics of the high mountain taxa of the genus Sideritis L. section Sideritis, subsection Fruticulosae Obón & D. Rivera (Lamiaceae)

Section Sideritis is an extremely diversified group which is formed mainly by species growing at low altitude. The group of high altitude taxa of this section is polyphyletic and has been taxonomically divided in different subsections in which these taxa appear normally associated with low-lying planitiary ones, the latter being the probable ancestors. The subsections comprising high altitude taxa are: subsect. Gymnocarpae , subsect. Fruticulosae; subsect. Hyssopifoliae , subsect. Luridae; subsect. Boigiae and subsect. Aranensis . Most of the high altitude taxa of section Sideritis are endemics with small distribution areas; they are incompletely known and threatened with extinction. The upper altitude limit of this section is attained at over 3000 m in Sierra Nevada (Spain) by Sideritis glacialis Boiss., which has been included in subsect. Fruticulosae Obón and D. Rivera. The taxonomy of this complex group of high altitude endemic taxa of Sideritis subsection Fruticuhsae is discussed on the basis of macro-morphological and micro-morphological characters. These are used in identification keys and for analysis of dissimilarity. The different habitats, allied species and plant communities are described. The following taxa are recognized: Sideritis glacialis subsp. glacialis, S. glacialis subsp. vestita, S. glacialis subsp. virens and S. glacialis subsp. fontquerii .     

Paeonia rotundiloba (D.Y.Hong) D.Y.Hong: A new status in tree peonies (Paeoniaceae)

Paeonia decomposita Hand.-Mazz.subsp,rotundiloba D.Y.Hong is here raised to the specific rank,P.rotundiloba (D.Y.Hong) D.Y.Hong,based on its distinctiveness of four diagnostic characters:number of carpels;height of disk; number of leaflets of the lower leaves,and shape of the terminal leaflets; and unpublished molecular data.Paeonia rotundiloba differs distinctly from P.decomposita in having carpels mostly 3,less often 2 or 4,very rarely 5 (vs.almost always 5,very occasionally 4 or 3),disk 8-15 mm high (vs.4-9.6 mm),leaflets mostly 19-39(vs.29-63) in number,and ratio of length to width of the terminal leaflets 1.09-1.93 (vs.1.81-2.99).     

Systematic studies on Paeonia sect. Moutan DC. based on RAPD analysis

Plants in Paeonia sect. Moutan DC., whose wild types are endemic to China, are deciduous subshrubs. Taxonomic treatments of most species in this section have long been in dispute. To address this question, both intraspecific and interspecific relationships of the species in this section were analyzed using RAPD markers. The dendrogram constructed by UPGMA showed that the accessions of the same species were always grouped together earlier than those of different species. The intraspecific similarity coefficients ranged from 0.60 to 0.90, grouping precisely those species of the same subsection together. Hence, the seven species under question can be well distinguished from each other. The similarity coefficient between P. delavayi and P. ludlowii was 0.60, and they were clustered in a clade. The similarity coefficients between P. jishanensis and the three species P. rockii, P. ostii, P. qiui, and between P. jishanensis and P. decomposita were both 0.48. These five species were clustered in another clade. These two clades corresponded well to Subsect. Delavayanae and Subsect. Vaginatae. Our results support the taxonomic treatment of Sect. Moutan re- cently proposed by Hong (1998,1997).