Microsatellite markers for assessing genetic diversity of the medicinal plant Paris polyphylla var. chinensis (Trilliaceae)

Paris polyphylla var. chinensis is a perennial herb with medicinal properties that is widely used in traditional Chinese medicine. However, this plant has been on the edge of extinction during the last few decades because of excessive deforestation based on the intense ethnopharmaceutical interest. We isolated 12 microsatellite loci from a (CT)(n)-enriched genomic library of P. polyphylla var. chinensis. The polymorphism at each locus was analyzed by screening 30 individuals from a natural population. The number of alleles ranged from 2 to 5. The observed and expected heterozygosities ranged from 0.000 to 0.467, with a mean of 0.247, and from 0.383 to 0.662 with a mean of 0.537, respectively. Six loci (Pp1, Pp3, Pp6, Pp7, Pp9, and Pp12) were found to significantly deviate from Hardy-Weinberg equilibrium. This may be due to the small population size, inbreeding or null alleles. Five of the pairwise comparisons (Pp1 and Pp4, Pp2 and Pp5, Pp2 and Pp9, Pp2 and Pp12, Pp11 and Pp12) exhibited significant linkage disequilibrium (P < 0.05). We conclude that these microsatellite markers will be useful for population genetic studies of P. polyphylla var. chinensis.     

Microsatellite loci from Taxus chinensis var. mairei (Taxaceae), an endangered and economically important tree species in China

Taxus chinensis var.mairei is an endangered and economically important tree species in China.Four polymorphic microsatellite markers were developed and characterized using the fast isolation by amplified fragment length polymorphism (AFLP) of sequences containing repeats (FIASCO) protocol.Polymorphism of each locus was quantified for five natural populations from Hubei Province,China.The number of alleles per locus varied from 6 to 24.The ranges of observed and expected heterozygosity were 0.493-1.000 and 0.440-0.845,respectively.These are the first microsatellites reported for the family of Taxaceae and will be helpful for the ongoing population and conservation genetics research of Taxus chinensis var.mairei.     

Isolation and characterization of microsatellites in the woody shrub, Banksia sphaerocarpa var. caesia (Proteaceae)

Microsatellite markers were developed for the Australian bird-pollinated woody shrub Banksia sphaerocarpa var. caesia to study gene flow among populations in a highly fragmented landscape. Eight loci were developed, and in a sample of 40 individuals from one population, the number of alleles per locus ranged from five to 21 and observed heterozygosities ranged from 0.385 to 0.914. All eight loci showed independent inheritance. Analysis of open-pollinated progeny arrays confirmed Mendelian inheritance at seven loci, while null alleles were suspected at the remaining locus.     

南方红豆杉丛枝菌根(AM)的研究

研究了南方红豆杉根部丛枝菌根真菌(AMF)侵染情况、菌根形态结构以及根际土中AMF孢子的种类与数量.结果显示:南方红豆杉可与AMF形成典型的丛枝-泡囊型菌根,侵染率在71.2%~94.4%,但是历山、蟒河自然保护区的侵染强度优于人工栽培区;在南方红豆杉根际土中共分离鉴定出5种AMF,无梗囊霉属1种、球囊霉属4种,分别是:光壁无梗囊霉、地表球囊霉、地球囊霉、缩球囊霉、明球囊霉,其中光壁无梗囊霉为优势种;南方红豆杉的根由表皮、外皮层、内皮层、中柱组成,AMF只侵染表皮层、内皮层,不能侵染中柱.这为将来利用AMF接种技术进行南方红豆杉的繁殖、移植栽培以及紫杉醇的积累等研究提供了理论依据.     

厚壳桂新微卫星体分子标记的开发与应用

生境破碎化是导致全球生物多样性危机的主要原因,即使常见物种也会受到很大影响.厚壳桂(Cryptocarya chinensis)是我国南亚热带季风常绿阔叶林群落演替顶极物种,由于这一森林植被的破坏而呈片断化分布.在以往的研究中,我们利用筛选到的一些微卫星体对其遗传多样性研究后发现其可能以无性生殖为主.为进一步证实这一发...     

PERMANENT GENETIC RESOURCES: Twelve new microsatellite markers for the Chinese shrimp Fenneropenaeus chinensis

Twelve new microsatellite markers were isolated by sequencing random clones from a genomic library of Fenneropenaeus chinensis. The number of alleles per locus ranged from five to 15. The polymorphism information content ranged from 0.568 to 0.898, and observed and expected heterozygosities from 0.344 to 0.882 and from 0.691 to 0.915, respectively. All loci except one conformed to Hardy-Weinberg equilibrium. These markers, described here for Chinese shrimp, will be further used to analyse the species' population genetics.     

Isolation and characterization of nine polymorphic microsatellite loci in the endangered shrub Disanthus cercidifolius var. longipes (Hamamelidaceae)

Nine polymorphic microsatellite loci were isolated and characterized from an AC-enriched genomic library of Disanthus cercidifolius var. longipes. Microsatellite polymorphism was investigated using 24 individuals from one natural population. The observed number of alleles per locus ranged from two to four. Observed and expected heterozygosities ranged from 0.17 to 0.92 and from 0.16 to 0.72, respectively. These polymorphic microsatellite loci provide useful tools for the ongoing population genetic studies of D. cercidifolius var. longipes.     

Isolation and characterization of microsatellite loci from Weigela coraeensis (Caprifoliaceae)

Weigela coraeensis var. coraeensis is a deciduous shrub species distributed in Japan on the mainland, Honshu, whereas its variety W. coraeensis var. fragrans is endemic to the Izu Islands located south of Honshu. We isolated eight polymorphic microsatellite loci from the species and characterized these loci for 20 individuals from a population in Honshu. The number of alleles per locus ranged from 7 to 15 and the observed and expected heterozygosities ranged from 0.60 to 0.90 and from 0.65 to 0.90, respectively. These eight polymorphic microsatellites will be useful for examining intraspecific genetic differentiation in W. coraeensis.     

南方红豆杉天然林群落结构特征研究

本文对福建三明陈大镇南方红豆杉天然林群落进行研究,测定了该群落11种主要树种的重要值,分析了该群落的种类组成、物种多样性、群落结构及其动态。结果表明,人为干扰较轻的南方红豆杉天然林群落比较稳定,天然更新良好,处于旺盛生长阶段。     

南方红豆杉无性系种群结构和动态研究

研究了元宝山南方红豆杉克隆种群的结构和动态．结果表明,种群S1～S6级个体均存在,S1、S2级个体占总个体数的64％．从元宝山南方红豆杉种群的大小级结构图看,结构呈较规则的金字塔形;南方红豆杉无性系幼树幼苗的贮备较为丰富,种群的年龄结构呈较规则的金字塔形,存活曲线近于倒J形;种群大小级动态指数Vpi=38．26％,考虑外部干扰时,Vpi′：6．38％>0,表现为稳定型种群．南方红豆杉种群更新的主要途径是无性繁殖,种群的数量动态主要受克隆内部调节机制以及外部环境的影响．生存分析结果表明,种群幼树幼苗的生存状态不稳定,死亡的可能性很大,而成年个体的生存状态则较为稳定．     

Isolation and characterization of microsatellite markers for Thujopsis dolabrata var. hondai (Cupressaceae)1

? Premise of the study: Microsatellite markers were developed for the endemic Japanese species Thujopsis dolabrata var. hondai for studies on forest ecology and tree breeding. ? Methods and Results: We characterized 10 dinucleotide microsatellite loci by screening primers developed using a simple sequence repeat-enriched library. The number of alleles per locus ranged from eight to 44 with an average of 20.3. The observed and expected heterozygosities ranged from 0.326 to 0.854 and from 0.670 to 0.976, respectively. ? Conclusions: The development of these markers can be used to assess useful genetic information for ecological studies and tree breeding in T. dolabrata var. hondai.     

元宝山南方红豆杉种群结构——Ⅱ、高度结构

在资源调查的基础上选择样地 ,采用样方调查法研究红豆杉种群高度结构 ,结果表明南方红豆杉种群是通过茎干萌生幼苗 (构件 )来延续和扩大的 ,其结构为塔型 ,存活曲线为中间微凸而近于直线 ,是稳定型种群结构。幼苗幼树级占 95.3% ,其次为高度 9.1～ 11m的成年树级 ,占 1.6 %。为了扩大和延续 ,南方红豆杉种群通过大量萌生 (繁殖 )构件并进行自我调节 ,使幼树幼苗有较高的死亡率 ;种群个体进入优势层后 ,高度生长缓慢 ,而径向生长加快 ,因此 9.1～ 11m级数量增多。种间种内竞争的结果形成南方红豆杉种群世代不同大小个体的高度结构。南方红豆杉种群高度与胸径的关系为幂函数相关关系 ,高度与年龄 (<6 0 a)的关系为线性相关关系     

Eight PCR primers to amplify EST‐linked microsatellites in the Eastern oyster,Crassostrea virginica genome

Twenty‐four microsatellite repeat sequences were identified by screening a total of 4446 eastern oyster, Crassostrea virginica, expressed sequence tags. Polymerase chain reaction primers were designed to amplify 12 of these loci. After optimizing reaction parameters, eight loci showed high variability with consistent amplification that could be scored unambiguously. Ninety two C. virginica from the James River, VA, were genotyped at these loci. Number of alleles per locus ranged from 11 to 53, expected and observed heterozygosities ranged from 0.69 to 0.97, and from 0.30 to 0.99, respectively. Discrepancies between expected and observed heterozygosities were common and likely caused by null alleles.     

Isolation and characterization of 12 polymorphic microsatellite markers for the frog Pelophylax hubeiensis

Twelve polymorphic microsatellite loci were isolated from an (AC)n- and (AG)n-enriched DNA library for the endemic Chinese frog Pelophylax hubeiensis (Ranidae). The number of alleles per locus ranged from two to eight, with a mean of 5.17. The observed and expected heterozygosities ranged from 0.226 to 0.839 and from 0.204 to 0.826, with means of 0.568 and 0.656, respectively. No significant linkage disequilibrium was detected among these loci. However, two significant deviations from HWE were discovered at loci Pehu-11 and Pehu-12 (P<0.05). MICRO-CHECKER tests showed that null alleles could be present at locus Pehu-12. These polymorphic microsatellite loci can be employed for exploring mating mechanisms, population genetic structure and other relevant genetic investigations of P. hubeiensis.     

Isolation and characterization of polymorphic microsatellite loci from Brandt's voles (Lasiopodomys brandtii)

Ten polymorphic microsatellite loci from Lasiopodomys brandtii have been isolated and characterized. Two to 11 alleles per locus were detected from 52 Brandt's voles samples collected from a single population. Expected heterozygosities ranged from 0.406 to 0.840. For the majority of loci observed heterozygosities were similar to or greater than the expected heterozygosity. One locus pair appeared to be in linkage disequilibrium. The microsatellite markers will enable the studies of genetic diversity, population structure and relatedness in this species, and perhaps in closely related species of vole.     

南方红豆杉毛状根诱导体系的建立及毛状根中 紫杉醇的分离纯化研究

目的:研究发根农杆菌对南方红豆杉转化的转化体系和毛状根中紫杉醇的提取纯化工艺;方法:采用发根农杆菌对南方红豆杉不同外植体进行诱导,并对影响毛状根转化的因素进行分析。采用超临界CO2流体萃取法、硅胶柱色谱法以及高效液相法对毛状根中的紫杉醇进行分离提纯;结果:成功构建了南方红豆杉毛状根诱导体系。其中农杆菌的种类、外植体类型、预培养和共培养时间、培养基中激素浓度均影响毛状根的转化率。毛状根经高压纸电泳检测显示表达冠瘿碱。毛状根中紫杉醇经超临界CO2法提取及硅胶柱色谱法分离提纯,采用高效液相法分析显示其纯度可达到98%。结论:发根农杆菌诱导南方红豆杉产生的毛状根中可产生紫杉醇,可作为紫杉醇的主要来源。     

Microsatellite markers for Spergularia media (L.) C. Presl. (Caryophyllaceae) and their cross-species transferability

We developed 11 polymorphic microsatellite markers for Spergularia media (Caryophyllaceae), a perennial halophyte of coastal salt meadows and continental areas of western Eurasia. The number of alleles per locus observed in a single population of 20 individuals from the German North Sea coast ranged from 3 to 20. Observed and expected heterozygosities ranged from 0.200 to 0.850 and from 0.278 to 0.936 respectively. Observed heterozygosities were lower than expected heterozygosities at all loci, presumably as a consequence of inbreeding. All markers cross-amplified in the closely related S. salina. Of these, nine were polymorphic in the heterologous species. Only two primer pairs generated PCR products in S. diandra.     

不同产地南方红豆杉枝叶的紫杉醇含量及其指纹图谱的比较分析

采用HPLC法对来源于6个产地(陕西周至和汉中;甘肃天水;安徽胡乐、聂家山和仙寓山)的12份野生南方红豆杉[ Taxus chinensis var.mairei( Lemée et Lévl.) Cheng et L.K.Fu]枝叶中的紫杉醇含量进行了比较分析,并对不同产地南方红豆杉指纹图谱的相似度进行了比较和特征指纹图谱的构建.结果表明:不同产地南方红豆杉枝叶中紫杉醇的含量有明显差异,来源于陕西和甘肃的枝叶样品中紫杉醇含量明显高于来源于安徽的样品,以产自陕西周至的枝叶样品中紫杉醇含量最高(0.019 3 mg·g-1).各样品的指纹图谱均有12个共有峰,其中9号共有峰的保留时间(32.75 min)与紫杉醇标准品一致,被确定为南方红豆杉指纹图谱的参照峰(s).各共有峰相对保留时间的RSD值均低于1％,但相对峰面积的RSD值均高于40％,说明各样品共有峰的保留时间稳定,但相对含量差异较大.除来源于安徽聂家山的2个样品外,其他样品的相似度值均在0.8以上,说明各样品间指纹图谱的相似度较高.根据12个样品的指纹图谱建立了南方红豆杉枝叶的特征指纹图谱,可用于南方红豆杉的鉴定及质量控制.由测定结果推测:不同产地南方红豆杉枝叶中紫杉醇含量的差异可能与生长环境的差异有关.     

Inoculations with Pseudomonas fluorescens and Bacillus cereus affect the soil enzyme activity,growth and rhizosphere microbial diversity of Taxus chinensis var. mairei

Plant and Soil - Taxus chinensis var. mairei is a rare tree endemic to China. Pseudomonas fluorescens CLW17 and Bacillus cereus CLY07 were isolated from the T. chinensis var. mairei rhizosphere and...     

濒危植物南方红豆杉种群生命表及谱分析

Based on the investigation in Longxi Mountain National Nature Reserve and the theory of survival analysis, a static life table of Taxus chinensis var. mairei population was worked out, the curves of its survival rate, mortality rate and killing power were drawn, and the population dynamics was analyzed by spectral analysis. The results showed that the survival curve of the population appeared to be a type of Deevey-Ⅲ, and the high mortality of seeding was one of the important reasons which caused Taxus chinensis var. mairei to be endangered. The spectral analysis of the population showed that there was a marked periodic regularity in the process of natural regeneration of Taxus chinensis var. mairei.