基于AFLP分子标记的桂花品种核心种质的构建

用100个桂花品种荧光AFLP分子标记信息构建核心种质。利用获得的指纹信息,运用UPGMA聚类取样法,采用Kimura 2-parameter遗传距离,多次聚类随机抽样。结果表明:(1)8对引物共获得514条带,平均每对引物获得64条带。(2)从100个桂花品种中筛选了30个样本的核心种质。(3)比较核心种质和全部种质的Shan-non-Wiener指数(H′)和Simpson指数(D),t检验值均说明核心种质的遗传多样性指数与全部种质遗传多样性没有明显差异,表明所构建的核心样品能够很好地保留原始100个桂花品种的遗传多样性。     

江西全南古桂和野生桂花资源调查

桂花[ Osmanthus fragrans(Thunb.)Lour.]为木犀科(Oleaceae)木犀属(Osmanthus Lour.)植物,是中国传统十大名花之一,已有2 500余年的栽培历史.桂花是一个长寿树种,在中国很多地方都有树龄上百年甚至上千年的古桂存在[1]18,[2].中国是世界上桂花古树保存最多、分布最广、最集中的国家[1]20.古桂不仅具有文物价值和旅游价值,还具有重要的科研价值,是研究古气候、古环境、古树种质资源以及新品种培育的绝好材料[3].     

桂花品种资源的遗传多样性分析

根据桂花(Osmanthus fragrans)品种的形态特征, 结合AFLP分子标记, 对部分桂花栽培品种进行了遗传多样性分析。结果表明, 桂花品种之间存在着较为丰富的遗传多样性, AFLP分子标记检测到的多态性条带占总扩增条带的57.46%。 根据桂花品种的主要性状特征, 利用数值分类法对其进行分类, 并用UPGMA法对AFLP结果进行聚类分析, 结果均显示桂林地区的桂花品种存在明显的区域性, 花色可以作为重要的分类标准, 同时对桂花品种的分类系统进行了探讨。     

桂花品种资源的遗传多样性分析

根据桂花（Osmanthus fragrans）品种的形态特征,结合AFLP分子标记,对部分桂花栽培品种进行了遗传多样性分析。结果表明,桂花品种之间存在着较为丰富的遗传多样性,AFLP分子标记检测到的多态性条带占总扩增条带的57.46%。根据桂花品种的主要性状特征,利用数值分类法对其进行分类,并用UPGMA法对AFLP结果进行聚类分析,结果均显示桂林地区的桂花品种存在明显的区域性,花色可以作为重要的分类标准,同时对桂花品种的分类系统进行了探讨。     

Characterization of microsatellites in wild and sweet cherry (Prunus avium L.)--markers for individual identification and reproductive processes.

Nuclear microsatellites were characterized in Prunus avium and validated as markers for individual and cultivar identification, as well as for studies of pollen- and seed-mediated gene flow. We used 20 primer pairs from a simple sequence repeat (SSR) library of Prunus persica and identified 7 loci harboring polymorphic microsatellite sequences in P. avium. In a natural population of 75 wild cherry trees, the number of alleles per locus ranged from 4 to 9 and expected heterozygosity from 0.39 to 0.77. The variability of the SSR markers allowed an unambiguous identification of individual trees and potential root suckers. Additionally, we analyzed 13 sweet cherry cultivars and differentiated 12 of them. An exclusion probability of 0.984 was calculated, which indicates that the seven loci are suitable markers for paternity analysis. The woody endocarp was successfully used for resolution of all microsatellite loci and exhibited the same multilocus genotype as the mother tree, as shown in a single seed progeny. Hence, SSR fingerprinting of the purely maternal endocarp was also successful in this Prunus species, allowing the identification of the mother tree of the dispersed seeds. The linkage of microsatellite loci with PCR-amplified alleles of the self-incompatibility locus was tested in two full-sib families of sweet cherry cultivars. From low recombination frequencies, we inferred that two loci are linked with the S locus. The present study provides markers that will significantly facilitate studies of spatial genetic variation and gene flow in wild cherry, as well as breeding programs in sweet cherry.     

Isolation and characterization of 11 microsatellite loci from Camellia sinensis in Taiwan using PCR-based isolation of microsatellite arrays (PIMA)

We report 11 novel microsatellite primer pairs for the wild tea, Camellia sinensis (L.) O. Kuntze forma formosensis Kitamura. These simple sequence repeat markers were tested in 24 samples collected from wild tea populations, and in cultivars and C. japonica. The number of alleles ranged from 4 to18. The expected (H _E) and observed (H _O) heterozygosity were 0.687–0.946 and 0.042–0.792, respectively. All loci were significantly deviated from Hardy-Weinberg expectations due to the heterozygote deficiency, indicating a dramatic loss of genetic polymorphisms in the rare species. Significant LD was discovered in most loci. These primers may provide a tool for understanding demography and population structure in wild tea.     

基于SRAP和TRAP标记的河北野生狗牙根种质遗传多样性分析

以SRAP和TRAP 2种标记技术对36份狗牙根材料的遗传多样性及亲缘关系进行了分析,其中包含34份河北省野生狗牙根种质资源。分别由238对SRAP和85对TRAP引物组合中筛选获得具有多态性的SRAP和TRAP引物组合各10对,PCR扩增总条带分别为186和161条,多态性条带156和132条,平均每对引物扩增出多态性条带各15.6和13.2条,多态性位点比率分别为83.4%和81.0%。2种标记合并进行聚类分析,所有供试的36份狗牙根材料遗传相似系数GS=0.519~0.983,平均为0.7。当GS=0.68时,可将36份供试材料分为4个类群。本研究结果表明河北野生狗牙根种质资源存在较丰富的遗传多样性,可为种质资源保护和选育优良狗牙根新品种提供科学依据。     

菰核基因组SSR引物的开发及其在稻族植物中的通用性检测

利用数据库中已有的部分菰(Zizania latifolia Turcz.)核基因组序列,采用in silico方法开发其SSR引物,并选取我国不同纬度的5个菰野生种群,对合成的64对引物进行筛选。结果显示:64对引物中有15对至少在一个种群中表现出多态性;共发现84个等位基因,每个位点平均有5.6个等位基因。在5个种群中,观察杂合度为0.000~0.941,预期杂合度为0.072~0.625。种群间的基因流(Nm=0.576)水平较低导致了种群间表现出较高的遗传分化(FST=0.432)。进一步对稻族其他物种的通用性检测发现,15个多态位点中,有8个位点在亚洲栽培稻(Oryza sativa L.)中得到扩增,有9个位点在普通野生稻(O.rufipogon Griff.)中得到扩增。     

基于SSR分子标记分析云南月季种质资源亲缘关系(英文)

利用简单重复序列SSR(simple sequence repeat)标记技术对48份月季种质资源(包括14份野生种、20份古老月季品种和14份栽培品种)的遗传多样性和亲缘关系进行了分析.结果表明,(1)18对SSR引物在18个SSR位点上共检测到160个等位基因,每一位点的等位基因变幅为6～13个,平均8.9个,材料间遗传相似系数变化范围为0.157 8～0.754 9,表明在分子水平上云南月季种质资源具有丰富的遗传多样性.(2)聚类分析结果显示,在相似系数为0.44处,可将 14个野生种明显分为6个组,这与植物形态学分类结果上基本一致;在遗传相似系数为0.40水平上将48份供试材料分为八大组群.(3)亲缘关系分析结果显示,5个野生种和所有古老月季品种与大多数栽培品种的亲缘关系较近.     

Nuclear and chloroplast SSR markers in Paeonia delavayi (Paeoniaceae) and cross-species amplification in P. ludlowii

? Premise of the study: Microsatellite primers were developed for Paeonia delavayi and P. ludlowii (Paeoniaceae) to study their population genetics and phytogeography. ? Methods and Results: Nine polymorphic nuclear microsatellite loci were isolated from an enriched library of P. delavayi and primers were designed. The number of alleles per locus ranged from two to 16; the observed and expected heterozygosities ranged from 0.014 to 0.687 and 0.042 to 0.875, respectively. Six polymorphic chloroplast microsatellite loci were identified in P. delavayi and primers were provided. The number of alleles per locus ranged from two to six and the polymorphic information content ranged from 0.08 to 0.716. Both nuclear and chloroplast primers were successfully applicable to P. ludlowii. ? Conclusions: The markers developed here will facilitate analyses of genetic diversity, population genetic structure, phytogeographical patterns, and conservation for P. delavayi and P. ludlowii.     

Isolation and characterization of new microsatellite markers in mango (Mangifera indica)

Six microsatellite loci that were isolated from a microsatellite‐enriched genomic library of mango (Mangifera indica) along with their specific primer sets were each characterized by using 36 cultivars collected mainly in Thailand. The observed and expected heterozygosities ranged from 0 to 0.83 and from 0.29 to 0.73, respectively. The number of putative alleles are two to six. Three of the six alleles have frequencies of over 75%. The high frequency may be attributed to the bias in the origin of cultivars. Among 36 mango cultivars tested, 29 cultivars showed a unique pattern by six primer sets, whereas seven cultivars cannot be identified because of genotype similarities. This suggests the potentials for identification of mango cultivars by microsatellite markers.     

Studies on Paeonia cultivars and hybrids identification based on SRAP analysis

Plants of Paeonia are valuable for their ornamental and medicinal values. Genetic relations and hybrids identification among different sections of Paeonia were studied using sequence related amplified polymorphism (SRAP) markers. A total of 29 cultivars including 2 intersectional hybrids, 13 sect. Moutan and 14 from sect. Paeonia were used. A total of 197 bands were produced using 24 primer combinations, among which 187 bands showed polymorphism. From the bands amplified, we can identify the peony cultivars using unique SRAP markers and specific primer combinations. Fourteen peony cultivars were distinguished among each other by using totally 35 SRAP markers, which were generated by 16 primer pairs. Two specific primer pairs of Me8/Em8 and Me8/Em1 can be used to identify cultivars from different sections. The mean genetic similarity coefficient (GS), the gene diversity (GD), and the Shannon's information index of peony cultivars were 0.45, 0.19 and 0.32, respectively. Both UPGMA (unweighted pair-group method of arithmetic average) dendrogram and PCA (principle component analysis) analysis showed clear genetic relationships among the 29 peony cultivars, and within section and its intersectional hybrids. The above results are valuable for estimating and analyzing genetic background of Paeonia, parent selection in crossing breeding programs, molecular marker assisted selection (MAS) breeding for further germplasm innovation programs.     

Identification and characterization of microsatellite markers derived from expressed sequence tags (ESTs) of radish (Raphanus satvius L.)

Radish, Raphanus satvius, is a very important commercial crop in the world, especially in East Asian countries. In this study, radish microsatellite markers have been developed for the first time from expressed sequence tags (ESTs). A total of 8 primer pairs were able to produce clear amplifications, respectively, which gave 2–5 polymorphic loci between 43 cultivars of Japanese radish. Observed and expected heterozygosity ranged from 0 to 0.4186, and from 0.1779 to 0.6205, respectively. Polymorphic information content ranged from 0.171 to 0.575. The eight simple sequence repeats were also polymorphic when tested by wild radish (Raphanus raphanistrum). Deviation from Hardy–Weinberg equilibrium was analysed in three populations, and no linkage disequilibrium was observed. It is anticipated that these newly developed microsatellite markers can advance further genomic analyses. This accomplishment may in turn accelerate and simplify radish breeding programs.     

利用RAPD研究桂林桂花品种间的亲缘关系

采用随机扩增多态性DNA(RAPD)技术,从100个随机引物中筛选出扩增效果较好的20个引物,分 析桂林市23个桂花品种的基因组多态性。20个随机引物共检测到193个位点,其中多态位点114个,占 59.1%。并进行了聚类分析,构建出树状聚类图,将这些品种划分为4个品种群,与传统分类学结果一致。结 果表明,以基因型而不是以表现型为基础,分析桂花品种间的区别是可能的。该技术为解决桂林市的桂花品 种分类问题提供了重要依据。     

Development and characterization of new microsatellite markers in <Emphasis Type="Italic">Panax</Emphasis><Emphasis Type="Italic">ginseng</Emphasis> (C.A. Meyer) from BAC end sequences

Panax ginseng, commonly known as Korean ginseng, is a valued source of herbal medicine in Korea and China. We have developed and characterized 35 microsatellite markers in P. ginseng from available BAC end sequences. Characterization of these 35 SSR primer pairs in 14 cultivars of P. ginseng showed 12 primer pairs to be polymorphic and 19 primer pairs to be monomorphic, while the remaining four primer pairs did not produce any product. The number of alleles amplified ranged from 4 to 8 per primer pair, with an average of six alleles per locus. The expected and observed heterozygosities ranged from 0.7500 to 0.9678 and 0.5645 to 0.7109, respectively. None of these loci deviated from Hardy–Weinberg equilibrium. All of the functional primer pairs of P. ginseng showed cross-species transferability with Panax quinquefolium. The cross-species transferable markers could be used for ginseng cultivar identification, for genomic studies, including mapping of specific trait QTL/genes, and for measuring conservation of ginseng.     

PERMANENT GENETIC RESOURCES: Microsatellite markers for the threatened Bliss Rapids snail (Taylorconcha serpenticola) and cross-amplification in its congener, T. insperata

We developed and tested microsatellite markers to investigate population structure of a threatened North American freshwater gastropod, Taylorconcha serpenticola. Of the 21 primer pairs that were evaluated, 11 were readily optimized and scored, providing amplification of 12 loci that were screened for 820 specimens from 29 populations. The number of alleles across 11 of these polymorphic loci ranged from three to 20 and the observed heterozygosity varied from 0.0061 to 0.7561. All loci yielded suitable amplification products in the second species of Taylorconcha (T. insperata) and three proved to be diagnostic for these congeners, demonstrating that these markers are also useful for species identification studies.     

Microsatellite primers for the endangered aquatic herb, Ottelia acuminata (Hydrocharitaceae)

? Premise of the study: Microsatellite primers were developed in the endangered aquatic herb, Ottelia acuminata, to characterize its genetic diversity and understand its population structure. ? Methods and Results: Eight polymorphic microsatellite markers were developed from two populations of O. acuminata in China. The number of alleles per locus ranged from one to 15; the observed and expected heterozygosities ranged from 0 to 0.885 and from 0 to 0.888, respectively, in the two populations. Selected loci also amplified successfully in O. sinensis. ? Conclusions: These microsatellite markers will facilitate further studies on the conservation genetics and evolutionary history of O. acuminata.     

Microsatellite loci for Gossypium davidsonii (Malvaceae) and other D-genome, Sonoran Desert endemic cotton species

? Premise of the study: Microsatellite primers previously developed for domesticated cotton (Gossypium hirsutum; tetraploid) were screened for their utility in investigating genetic structure and gene flow within G. davidsonii and five other wild, Mexican, D-genome cotton species (all diploid). ? Methods and Results: We screened 50 microsatellite primer pairs from the Cotton Marker Database, identifying 10 loci as polymorphic within G. davidsonii. In genotyping approximately 200 individuals from four populations, we found that the number of alleles per locus ranged from seven to 17, and mean observed and expected heterozygosity ranged from 0.145 to 0.492 and from 0.436 to 0.734, respectively. We genotyped six to 20 individuals from each of the remaining species, finding these 10 loci to cross-amplify in all cases and in most cases to be polymorphic. ? Conclusions: These markers may be useful for further investigation of population genetics of G. davidsonii and other wild D-genome cotton species.     

29份云南甘蔗创新种质的SSR遗传多样性分析和指纹图谱构建

该研究以16份甘蔗骨干亲本为参照,对29份云南甘蔗创新种质进行SSR指纹图谱构建和遗传多样性分析,以明确创新种质与16份亲本间的遗传基础和多样性水平。结果表明:6对引物共扩增出104条带,其中101条为多态性条带,多态性条带比例为97.25%;45份材料的遗传相似性系数为0.235 3~0.891 3,平均值为0.563 3;其中16份甘蔗骨干亲本的遗传相似性系数为0.301 6~0.755 6,甘蔗创新种质与甘蔗骨干亲本的特异条带比例为14∶1,涵盖了割手密、大茎野生种、斑茅和滇蔗茅等基因源。根据骨干亲本间的相似性系数范围,在相似性系数为0.43处,可将种质分为6大类群,亲缘关系相对较远,适宜作为种质间的杂交利用。通过引物区分效率分析,6对引物扩增的多态信息量为0.967 9~0.975 8,其中MSSCIR21引物区分效率最高,利用MSSCIR21和SMC1047HA引物组合构建了云南甘蔗创新种质标准指纹图谱,在相似性系数为0.85处即可区分所有种质,图谱的鉴别准确率为100%,每份资源都有唯一的指纹图谱,可将29份创新种质和16份骨干亲本区分鉴别出来。该研究能够为后续杂交利用、种质鉴定和知识产权保护提供依据。     

Sixteen novel microsatellite markers developed for Dendrocalamus sinicus (Poaceae), the strongest woody bamboo in the world

? Premise of the study: Microsatellite primers were developed in the strongest and most economically important bamboo species, Dendrocalamus sinicus, to investigate its genetic variability. ? Methods and Results: Using the Fast Isolation by AFLP of Sequences COntaining Repeats (FIASCO) protocol, 16 polymorphic primer sets were identified within 24 individuals from two geographic locations. The number of alleles per locus ranged from two to four, with a mean of 2.6. The observed and expected heterozygosities varied from 0 to 1.000 and from 0.311 to 0.754, respectively. The cross-amplification of the 16 primer pairs was tested in four other Dendrocalamus species, and nine to 13 (56.3-81.3%) primer pairs were successfully amplified. ? Conclusions: These markers will be useful for studies on the genetic diversity of D. sinicus, which may facilitate conservation of this species.