Microsatellite markers in the tree peony, Paeonia suffruticosa (Paeoniaceae)

? Premise of the study: Microsatellite primers were developed in the tree peony, Paeonia suffruticosa, to perform paternity tests as well as assignment to variety in special Austrian collections. ? Methods and Results: Using SSR-enriched libraries and EST-mining, 8 polymorphic primer sets were identified in Austrian collections of Paeonia sect. Moutan DC. The primers amplified di- and trinucleotide repeats with 2-6 alleles per locus. All primers also amplified in P. ostii, P. pontaninii var. trolloides, P. delavayi, and P. lutea, and in the herbaceous species P. peregrina and P. tenuifolia (Paeonia sect. Paeon). ? Conclusions: These results show the usefulness of primers in P. suffruticosa for population genetic studies and their ability to cross amplify in related taxa across the genus.     

Cytogeography and taxonomy of the Paeonia obovata polyploid complex (Paeoniaceae)

 The taxonomy of the Paeonia obovata polyploid complex in E-Asia has been controversial with treatments ranging from one species and two varieties to five species and numerous infraspecific taxa. Extensive observations in the field, analysis of morphological characters based on 32 populations sampled, and the examination of a large amount of herbarium specimens show that all the characters used by previous authors for distinguishing the species are variable between or even within populations: Leaf position, petal colour, stamen number (from 21 to 110 in the red-flowered and from 58 to 239 in the white-flowered form), filament and anther colour, style length as well as follicle number and position. Therefore, all these characters are of no value for species delimitation. However, an extensive chromosome survey and the analysis of morphological characters show that the ploidy level is basically correlated with the geographical distribution and with the indumentum on the lower leaf surface. Thus, the recognition of one species with two subspecies is proposed: Paeonia obovata subsp. obovata, mostly diploid, widely distributed in the north, east and south of the distribution range and glabrous to sparsely pubescent or hirsute on the lower leaf surface, and subsp. willmottiae, tetraploid, limited to the west and with leaves densely hirsute or pubescent below. Received October 30, 1998 Accepted June 21, 2000     

Paeonia anomala的核型研究

报道了Paeonia anomala L．的核型,澄清了新疆阿尔泰地区分布的该物种的染色体数目。分布于该地区的Paeonia anomala L．的核型组成：2n=2x=10=6m＋2sm＋2st。该类群核型与该属其它类群一致——2A型。在综合比较分析该属染色体参数以及核型不均一性参数,包括最长,最短染色体比（L1/Ln）和染色体不对称系数（CKOA）的基础上,我们发现该属三个组在核型上没有明显分化,仅在木本类群（Sect．Mouton DC．）和草本类群（Sect．Onaepia Lindley和Sect．Paeonia）之间存在微小差异。此外,作为二倍体类群,新疆阿尔泰地区分布的Paeonia anomala L．很可能是二倍体杂种,这将为研究被子植物的父母本同倍化杂交式物种起源提供一个很好的研究材料。     

川赤芍的化学成分研究

从川赤芍Paeonia anomala subsp·veitchii根皮的70%丙酮提取物中,分离鉴定了22个化合物,其中包括一个新的24,30位降常春藤皂苷三萜衍生物,命名为paeonenolide H(1)。化合物2,4,9,10为首次从该植物中分离得到。     

滇牡丹花色类群性状变异分析

对滇牡丹（Paeonia delavayi）10个花色类群的13个数量性状和5个质量性状进行了调查及统计分析,结果表明：（1）未发现所有花部性状均为同色的花色类群;（2）从花色与其它质量性状的连锁看,绿色类群均具绿色柱头,且是唯一存在直立花朵、绿色花丝、绿色花药的类群;（3）10个花色类群13个数量性状的平均变异系数范围为32．80％～50．56％,说明不同类群内个体问的性状稳定性差异较大;（4）10个花色类群平均表型分化系数为42．25％,表明滇牡丹10个花色类群表型变异主要来源于类群内,类群间的表型变异也很突出;（5）据花色演化趋势,初步推断云南省迪庆州中甸、德钦一带可能为滇牡丹的起源中心。     

Molecular phylogenetic evidence for the origin of a diploid hybrid of Paeonia (Paeoniaceae)

There is growing evidence that hybridization not only by means of allopolyploidy but also at the homoploidy level was a major driving force of plant diversification. While allopolyploidy is known to be a common mode of speciation in Paeonia (Paeoniaceae), hybrid speciation at the diploid level needs further evaluation. Paeonia anomala was previously considered to be an interspecific hybrid but with an unknown ploidy level. In this study P. anomala is identified as a diploid (2n = 10). With increased sampling of populations and molecular markers, we showed that P. anomala is a homoploid hybrid that originated from a cross between P. veitchii and P. lactiflora. Five populations of P. anomala were sequenced for the following molecular markers: the matK gene and two intergenic spacers, psbA-trnH and rps16-trnQ, of the chloroplast genome; the internal transcribed spacers (ITS) of nuclear ribosomal DNA; and three low-copy nuclear genes, Adh1, Adh2, and Gpat. The populations of P. anomala were grouped together with P. veitchii on the ITS and Gpat phylogenies but with P. lactiflora on the chloroplast phylogeny. Sequence polymorphism was found at the Adh1 and Adh2 loci within individuals of P. anomala. These polymorphic sequences were grouped with P. veitchii and P. lactiflora, respectively. Phenetic analysis indicated that P. anomala is morphologically similar to P. veitchii. Phenotypic evolution resulting from the combination of two diverged genomes might have occurred primarily at the physiological level and allowed P. anomala to adapt to geographic regions different from those of its parents.     

Sex allocation under simulated herbivory in the generalist perennial herb Paeonia broteroi (Paeoniaceae)

Theories of optimal resource allocation in flowering plants postulate that allocation to sexual functions are balanced. While many studies have found such evidence in hermaphroditic species among flowers, plants or populations, or at different phenological stages, it has not been supported by other studies. This has been explained by differences among genotypes in ability to acquire resources, resource partitioning among traits unrelated to reproduction or strong selection to maintain positive genetic correlation among traits. I studied how herbivory affected resource allocation to sexual functions in the perennial herb Paeonia broteroi (Paeoniaceae) by measuring a number of floral traits in control plants and in experimental plants under simulated herbivory. The species shows very little plasticity in resource allocation between sexual structures and functions, and appears to be highly sensitive to alterations in the balance of resource acquisition and allocation, with an immediate outcome in terms of seed production and a mid-term response in terms of sprouting and flowering. Plants' ability to collect resources for growth and reproduction before their senescence at the end of the reproductive season, are compromised. This may limit their reproductive potential, the maintenance and growth of their populations, and may influence their demographic state and the plants' ability to respond to selective pressures by external biotic agents.     

Enzyme inhibition and radical scavenging activities of aerial parts of Paeonia emodi Wall (Paeoniaceae)

The ethanolic extract derived from aerial parts of an indigenous medicinal plant Paeonia emodi was screened for enzyme inhibition activities against Urease (jack bean and Bacillus pasteurii) and alpha-Chymotrypsin. The extract was also investigated for its radical scavenging activity using DPPH assay. The crude extract was found to possess significant enzyme inhibition activities against jack bean (74%) and Bacillus pasteurii (80%) urease and a moderate activity (54%) against alpha-Chymotrypsin. The extract also displayed excellent (83%) radical scavenging activity. On the basis of these results, the crude extract was subsequently fractionated into n-hexane, chloroform, ethyl acetate, n-butanol and water fractions and tested independently for the aforesaid activities. Significant inhibitory activity against urease enzyme was observed for the ethyl acetate, n-butanol and water fractions while the n-hexane and chloroform fractions were devoid of any such activity. In the alpha-Chymotrypsin enzyme inhibition studies the activity was concentrated into the ethyl acetate fraction. All the fractions displayed potent radical scavenging activity. The crude extract and fractions thereof were also subjected to total phenolic content determination. A correlation between radical scavenging capacities of extracts and total phenolic content was observed in the majority of cases.     

A revision of the salvia dorrii complex (Lamiaceae)

Expressions of theSalvia dorrii (Kellogg) Abrams complex, a member of subg.Audibertia Benth., are distributed over much of the western United States. The complex consists of two species,S. dorrii andS. pachyphylla Epling ex Munz.Salvia dorrii is divided into subsp.dorrii and subsp.mearnsii (Britton) McClintock, the latter endemic to central Arizona. Subspeciesdorrii is composed of four varieties: var.carnosa (Douglas ex Greene) Cronq., a large-leaved erect shrub of Washington and Idaho southward through Oregon into extreme north-central California: var.dorrii, a small-leaved erect shrub of southern Oregon and Idaho southward through the Great Basin of Nevada and western Utah to southeastern California and northern Arizona: var.pilosa (A. Gray) Strachan & Reveal, a small-leaved erect shrub differing from var.dorrii by pilose bracts and calyx, in southern California and western Arizona with disjunct populations in the Lahontan Basin of northwestern Nevada and northestern California; and var.clokeyi Strachan, var. nov., a low mat-forming subshrub of the high mountains of Clark Co., Nevada.Salvia pachyphylla ranges from southern California southward to the high mountains of Baja California Norte, Mexico, and eastward across extreme southern Nevada to (as a disjunct) northeastern Arizona. Each entity is described in detail, mapped and illustrated.     

Phylogenetic utility of the glycerol-3-phosphate acyltransferase gene: evolution and implications in Paeonia (Paeoniaceae)

The nuclear-encoded chloroplast-expressed glycerol-3-phosphate acyltransferase (GPAT) gene has been found to be single-copy in a number of angiosperm families. In this study we investigated the phylogenetic utility of the GPAT gene at the interspecific level using the genus Paeonia (Paeoniaceae) as an example. An approximately 2.3- to 2.6-kb fragment of the GPAT gene, containing a large intron of more than 2 kb, was amplified, cloned, and sequenced from 19 accessions representing 13 Paeonia species. The GPAT gene phylogeny inferred by parsimony analysis supported interspecific relationships that were previously unresolved, suggesting that large introns of low-copy nuclear genes are particularly informative in the resolution of close relationships at low taxonomic levels. Whereas the GPAT phylogeny is largely congruent with the previous phylogenetic hypothesis of Paeonia, it shows a significant discordance involving the paraphyly of section Paeonia. Given evidence of an ancient duplication and the subsequent silencing of one GPAT locus in P. anomala, this discordance is most likely the result of paralogy. Two distinct genomic clones containing partial GPAT genes were isolated from P. anomala. The GPAT sequence from one clone corresponded to the functional copy of the gene, and the second genomic clone was determined to contain a GPAT pseudogene. The insertion of a retrotransposon in an intron of this pseudogene may have been responsible for the silencing of this GPAT locus in P. anomala. This study suggests that, although it is unlikely that universal nuclear gene markers free from paralogy are usually available, low-copy nuclear genes can be very useful in plant phylogenetic reconstruction, especially at low taxonomic levels, as long as the evolutionary dynamics of the genes are carefully examined.     

牡丹试管苗玻璃化的研究

以MS为基本培养基,研究品种、琼脂、6-BA、大量元素、蔗糖、光照等因素对牡丹试管苗玻璃化的影响。结果显示：不同品种的牡丹试管苗在组织培养的过程中,出现不同程度的玻璃化,增加培养基中琼脂浓度、降低细胞分裂素6-BA的浓度、降低大量元素的浓度和增加光照强度或采用自然光照均可以有效地降低试管苗玻璃化。     

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).     

利用新生子叶制备牡丹染色体的研究

首次报道了利用牡丹(P．suffruticosa)种子的新生子叶制备染色体,同时比较了根尖染色体的核型,结果表明:牡丹子叶和根尖染色体核型数据一致,2种方法获得的染色体数均为2n=10,核型公式为2n=2x=10=6m 2sm 2st,全套染色体未见随体;2种方法获得的染色体形态结构基本相同。相对根尖而言,牡丹种子的新生子叶面积大,容易获得更多的试材,说明利用新生子叶制备染色体更适合遗传学实验研究。     

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.     

牡丹籽的化学成分研究

从毛茛科芍药属牡丹Paeonia suffruticosa Andr．种子甲醇提取物中分离得到13个化合物,通过理化性质及波谱分析鉴定为齐墩果酸（oleanolic acid,1）、12,13-dehydromicromeficacid（2）、常春藤皂甙元（hederagenin,3）、山奈酚（kacmpferol,4）、木犀草素（luteolin,5）、芹菜素（apigenin,6）、柯伊利素（chrysoeriol,7）、反式葡根素（trans-ε-viniferin,8）、顺式葡根素（cis-β-viniferln,9）、反式白藜芦醇（trans-resveratrol,10）、β-谷甾醇（β—sitosterol,11）、豆甾醇（stigmasterol,12）和β-胡萝卜苷（β-daucosterol,13）。所有化合物均首次从该植物的种子部位中得到。