Genetic structure of Tunisian natural carob tree (Ceratonia siliqua L.) populations inferred from RAPD markers

? Seven RAPD markers were used to assess the genetic diversity and structure of ten Tunisian natural Ceratonia siliqua L. populations from different geographic and bioclimatic zones.

? The species maintain a high diversity within population as estimated by the percentage of polymorphic loci and Shannon’s index (P% = 76.31, $\bar H'_{pop} = 0.569$ ). The range of variation between populations was large. Populations from the upper semi-arid bioclimates, with more continuous distribution area showed the highest level of variation.

? A high genetic differentiation among populations (Φ_ST = 0.250 and $\bar G_{ST} = 0.347$ ), as a result of population isolation was revealed. Nevertheless, the genetic structure is in accordance with bioclimate indicating that ecological factors also should influence differentiation. Populations from the sub-humid, upper semi-arid and mean semi-arid zones clustered together and were distinct from those of the lower semi-arid ones.

? Conservation strategy should be made according to the level of polymorphism within population and bioclimate.

     

Assessment of diversity among populations of Rauvolfia serpentina Benth. Ex. Kurtz. from Southern Western Ghats of India,based on chemical profiling,horticultural traits and RAPD analysis

Genetic, morphological and chemical variations of ten natural populations of Rauvolfia serpentina Benth. Ex. Kurtz. from Southern Western Ghats of India were assessed using RAPD markers reserpine content and morphological traits. An estimate of genetic diversity and differentiation between genotypes of breeding germplasm is of key importance for its improvement. Populations were collected from different geographical regions. Data obtained through three different methods were compared and the correlation among them was estimated. Statistical analysis showed significant differences for all horticultural characteristics among the accessions suggesting that selection for relevant characteristics could be possible. Variation in the content of Reserpine ranges from 0.192 g/100 g (population from Tusharagiri) to 1.312 g/100 g (population from Aryankavu). A high diversity within population and high genetic differentiation among them based on RAPDs were revealed caused both by habitat fragmentation of the low size of most populations and the low level of gene flow among them. The UPGMA dendrogram and PCA analysis based on reserpine content yielded higher separation among populations indicated specific adaptation of populations into clusters each of them including populations closed to their geographical origin. Genetic, chemical and morphological data were correlated based on Mantel test. Given the high differentiation among populations conservation strategies should take into account genetic diversity and chemical variation levels in relation to bioclimatic and geographic location of populations. Our results also indicate that RAPD approach along with horticultural analysis seemed to be best suited for assessing with high accuracy the genetic relationships among distinct R. serpentina accessions.     

濒危小灌木长叶红砂种群的遗传多样性

采用RAPD和ISSR2种分子标记对濒危小灌木长叶红砂5个种群的遗传多样性进行检测。18个RAPD引物和14个ISSR引物分别扩增出118和114个位点,多态位点比率(P)分别为88.98%和89.47%。在物种水平上,RAPD标记的结果为:Shannon’s信息多样性指数(I)为0.4656,Nei’s指数(H)为0.3303;ISSR检测的结果:I=0.4688,H=0.3083。2种分子标记均表明濒危小灌木长叶红砂具有较高的遗传多样性水平。Nei基因多样性指数表明,大部分遗传变异存在于种群内。RAPD分析发现86.22%的遗传变异发生在种群内;ISSR分析发现89.29%的遗传变异发生在种群内。种群间遗传变异低的主要原因是种群间存在较强的基因流(Nm)分别为3.0097和4.1787)。长叶红砂较高的遗传多样性水平与物种特性和对高胁迫环境的长期适应有关,濒危植物并不一定表现为遗传变异水平的降低。     

分子标记法和形态学特征对杏仁栽培种及野生种基因型的遗传关系构建的辨别

应用23个形态学特征,19个扩增片段长度多态性(AFLP)引物组合,80个随机扩增多态DNA(RAPD)引物和32个简单序列重复(SSR)引物对,比较三种分子标记法在29个杏仁栽培种和3个野生种遗传关系构建中的信息景和效率.根据预期杂合度的评价,与AFLPs和RAPDs相比,SSRs具有较高水平的多态性和较大的信息量.AFLPs预期朵合度值最低,但其辨别效率值最高,因为AFLPs能揭示每个反应中的大量条带,导致各种类型的多样性指数均较高.三种分子标记法对杏仁基因型的辨别效率均较高,只是SSRs无法辨别‘Monagha'和‘Sefied'杏仁基因型.三种分子标记法基因型相似性相关系数统计上显著,但SSR数据要低于RAPDs和AFLPs的值.尽管三种分子标记法树形图拓扑结构存在一些差异,但相似性水平均较高.SSRs、RAPDs和AFLPs的系统树图及其综合数据都能依据地理散布反映大多数栽培种的关系.AMOVA检测到每个地理组中栽培种和野生种的变异.辅助程序分析表明,实验所应用的标记物数量足以保证基因相似性估计的可靠性和标记法间的比较是有意义的.     

白桦种群间木材纤维性状的表型变异与分子标记的遗传多态性

以东北地区的5个白桦天然种群为研究材料,在木材纤维形态性状测定的基础上,采用ISSR、RAPD分子标记研究了5个种群的遗传变异以及种群间纤维形态性状与DNA分子标记的相关性.结果表明:白桦天然林成熟材(18～31年生)的木材纤维长度在种群间差异不显著,纤维宽度和长宽比差异显著,帽儿山种群的木材纤维形态性状最好,汪清最差;利用ISSR和RAPD 2种标记检测5个白桦种群的遗传多样性,所得的研究结果基本一致,多态位点比率最高的为辽宁新宾种群,其次是帽儿山和汪清,而塔河和金山屯最低;ISSR标记的AMOVA分析表明白桦种群间的遗传变异占总遗传变异的10.82%;RAPD标记的AMOVA分析表明白桦种群间的遗传变异占总遗传变异的12.94%,研究结果均显示白桦的遗传变异主要发生在种群内部.利用2种标记技术的种群聚类结果基本一致,这一结果与基于木材纤维性状的表型聚类基本一致, 而且种群间遗传距离和地理距离之间存在一定的相关性.     

Genetic diversity in Tunisian Crataegus azarolus L. var. aronia L. populations assessed using RAPD markers

? The genetic diversity of nine wild Tunisian Crataegus azarolus var. aronia L. populations from different bioclimates was assessed using RAPD markers.

? Eight selected primers generated a total of 105 bands, 81 of which were polymorphic. Shannon’s index (H′) ranged from 0.222 to 0.278 according to a population with an average of 0.245. The genetic variation within the species (H _SP = 0.423) was relatively low. A high differentiation (G _ST = 0.421) among populations coupled with a low level of gene flow (N _m = 0.472) were observed. The analysis of molecular variance (AMOVA) revealed also significant differentiation among populations (Φ_ST = 0.371), even at a low scale space. The majority of variation occurred within populations (63.31%). The Mantel test performed on genetic (Φ_ST) and geographic distance matrices among population pairs did not reveal an isolation by distance.

? Interpretation of Neighbour-joining tree based on Nei’s and Li’s genetic distance among individuals showed distinct population groupings. The UPGMA dendrogram based on Φ_ST values revealed two population sub-clusters, each including populations from different bioclimates and/or geographic regions.

? The low level of genetic diversity and the high genetic structure of populations resulted from genetic drift caused both by habitat fragmentation and the low size of populations.

? The high differentiation among populations and the similar low level of diversity within populations suggest that in situ conservation should interest all populations. The ex situ conservation should be based on the collection of seeds rather within than among populations because of the maximum of variation was revealed within populations.

     

云杉属树种天然群体DNA标记的国外研究进展

本文综述了自21世纪初以来DNA标记技术在国外云杉属树种中的应用情况,涉及到的树种以挪威云杉为主,多至十几种,标记种类主要包括AFLP、RAPD、RFLP、SSR、ISSR、STS、EST、VNTR和SNPs等。另外,对几种群体遗传参数进行比较分析,表明云杉天然群体遗传变异丰富,其中利用ISSR检测到白云杉天然群体多态位点百分率可达90%,利用RAPD检测到挪威云杉群体总遗传变异高至0.941;群体间存在遗传分化,产生了适应环境的变异,除一个利用mtVNTR研究所得结果外,其他研究均表明,云杉群体间遗传分化程度并不是很高,云杉群体变异主要表现为群体内变异。同时,对DNA标记在云杉属种质资源保存及遗传改良方面的前景进行了展望。     

Analysis of genetic relationship among Arbutus unedo L. genotypes using RAPD and SSR markers

The strawberry tree (Arbutus unedo L.) is an underutilized, drought tolerant, fire resistant species with a south western distribution in Europe, and with ecological and putative socio-economical impact in Portugal and Mediterranean countries. Our aim was to develop an appropriate set of molecular markers to enable genetic diversity to be assessed and to fingerprint Arbutus unedo genotypes for breeding and conservation purposes in Portugal. Twenty-seven trees from a broad geographic range were screened with 20 random amplified polymorphic DNA (RAPD primers) and 11 microsatellite markers (SSR). The RAPDs generated 124 bands, 57.3% of which were polymorphic, with an expected heterozygosity of 27%. We cross-amplified 11 SSR primers developed for Vaccinium spp., and 5 were found to be polymorphic in A. unedo, with 75% of expected heterozygosity, a number of alleles of 11.6, a null allele frequency of 7.6% and a polymorphic information content of 71%. Although the SSRs were more polymorphic and informative than the RAPDs, both markers displayed high genetic variability with the gathered data. No geographic pattern was observed in the genetic variation distribution based on both marker systems, and the lack of correlation between genetic and geographical matrices was confirmed by Mantel tests. Likely, no correlation was found between pairwise SSR and RAPD band-sharing matrices. These results and their implications on A. unedo breeding and conservation programs are discussed.     

Isozyme and RAPD polymorphisms in Heterobasidion annosum in Italy

Isozyme and random amplified polymorphic DNA (RAPD) polymorphisms were used to study variability in a group of 41 isolates from the Italian population of Heterobasidion annosum. The isolates belonged to the intersterility groups P and S, and particularly to the group that is most widely distributed in Italy, group F. Isozyme analysis was effective in identifying the three intersterility groups and revealed a high degree of genetic divergence within the P group isolates; the mannose phosphate isomerase (MPI-2) locus was diagnostic in the attribution of isolates to the more correlated F and S groups. RAPDs were detected following amplification by the polymerase chain reaction (PCR). 74 RAPD fragments, obtained through amplifications with eight primers, were scored. Isolates from the 3 intersterility groups were clearly divergent based on analysis of RAPD markers. However, a similarity index calculated for the isolates within the F population indicated a high uniformity of the isolates collected throughout the Italian peninsula.     

Molecular characterization and genetic relationships among most common identified morphotypes of critically endangered rare Moroccan species Argania spinosa (Sapotaceae) using RAPD and SSR markers

? The objective of this work is the molecular characterization of most common identified morphotypes of critically endangered rare Moroccan species Argania spinosa.

? Eighteen RAPD markers and twenty SSR markers have been assayed in 38 argan tree accessions from the three most commonly identified morphotypes: oval, spherical and spindle fruit types.

? A total of 140 polymorphic RAPD bands were detected out of 146 bands. The number of presumed alleles revealed by the SSR analysis ranged from 1 to 5 alleles per locus with a total number of 32 alleles identified. Results demonstrated an extensive genetic variability within the tested argan accessions. RAPDs presented a high level of polymorphism and greater information content than SSRs.

? Our results could indicate that Vitellaria paradoxa is genetically closer to argan than Manilkara huberi. In addition, the correlation between the clustering based on RAPD and SSR markers were in general low. The observed clustering could be better explained according to geographic proximity than morphotype. For this reason the traditional morphological characterization of the argan accessions in morphotypes (according to fruit phenotype) seem not to be in correlation to the real genetic background (genotype) of this specie. The implications of these results in the creation of effective germplasm core collection in argan have been also discussed.

     

Molecular characterization and marker based chemotaxonomic studies of Podophyllum hexandrum Royle

Detailed chemical studies and RAPD analysis were done in different populations of Podophyllum hexandrum collected from high altitude regions of North Western Himalayas. Random amplified polymorphic DNA (RAPD) analysis revealed a high degree of genetic diversity among the 12 collected accessions, attributed to their geographical and climatic conditions. HPLC analysis also revealed variation in the concentration of two major marker compounds which lead to the identification of a chemotype. The study demonstrated that RAPD and chemical markers are very useful tools to compare the genetic relationship and pattern of variation among such prioritized and endangered medicinal plants.     

Variation in Heterobasidion annosum detected by Random Amplified Polymorphic DNAs

The production of Random Amplified Polymorphic DNAs (RAPDs) by Polymerase Chain Reaction (PCR) was used to detect variation in the isolates of Heterobasidion annosum with various geographical origins. Specific RAPD products were detected for each of the intersterility groups: the European S, F, P and the North American S group. There was considerably more polymorphism found among European S than P isolates. The analysis of RAPDs was shown to be a simple and a fast way to generate DNA markers specific to the previously established intersterility groups and thus useful for diagnostic purposes.     

美国山核桃群体遗传多样性的RAPD分析

运用随机扩增多态性DNA（RAPD）标记技术，采用Operon公司的10种随机引物，以采自美国东南部、北部、西部和我国江苏、浙江、湖北、湖南、云南等地的9个美国山核桃栽培群体45个单株的种子为材料，检测出多态位点42个。以这些多态位点为依据，计算了估测了多态性位点百分比、Shannon表型多样度指数、Virk群体内遗传距离和Nei氏群体间遗传距离，并用它们作为参数进行群体遗传多样性的RAPD分析。结果表明，参试的美国山核桃品种群体遗传变异明显，且群体间差异大于群体内差异，我国引种的群体遗传差异大于原产地美国的群体遗传差异。     

福建光皮桦野生种群遗传变异及其与生境的关系

采用表型性状测定结合RAPD标记,研究福建境内8个光皮桦野生种群的遗传变异,探讨遗传变异与环境因子间的相关性.结果表明:光皮桦野生种群在DNA和表型水平上的遗传变异基本一致,即约2/3的变异存在于种群内,1/3存在于种群间;叶、果序和种子3类性状的变异系数(CV)分别为0.063 2,0.070 6和0.069 3,各类性状的形状指数(长宽比)的变异系数均比单个性状小;种群平均多态位点百分比(PPL)、Nei's基因多样性(h)和Shannon信息指数(I)分别为65.27%,0.246 1和0.362 8.相关分析表明:表型性状间及其与经纬度、年均温、年日照时数、海拔及有机质等环境因子间存在显著或极显著相关,与各环境因子相关的主要表型性状有叶长宽比、侧脉数、叶基角、种翅宽、种子千粒质量和果序长宽比等;PPL,h和I与年日照时数呈显著相关,与经纬度、年降水量呈负相关;种群间的遗传距离与地理距离相关不显著,而与年均温分异显著相关.     

Effect of Regeneration Method on RAPD-Based Genetic Variation of Cyclobalanopsis glauca (Fagaceae)

Cyclobalanopsis glauca is a dominant species of evergreen broad-leaved forests in mainland China. This study compares the genetic variation of an artificially regenerated population with its donor population and two other wild populations, by using RAPD markers. A total of 74 clear, reproducible bands were scored for 12 RAPD primers; 72 were polymorphic (P = 97.3%). AMOVA revealed that most genetic variation was within populations and only 10.35% was among populations. Various measures indicated that there is no difference in genetic diversity between the planted and the original populations. Φ_ST between the planted offspring population and the donor population was larger than those between the planted and other two natural populations, indicating that artificial regeneration might lead to biased genetic composition, given that temporal differentiation is usually lower than spatial differentiation. This divergence may be due to unequal seed production among the maternal individuals and viability differences among seeds.     

An analysis of the pattern of genetic variation in Vitellaria paradoxa using RAPD markers

Vitellaria paradoxa C.F.Gaertn. is one of the most economically and socially important tree species in the Sudano-Sahelian region. Little is known of the pattern of variation within its natural range. Eight populations covering most of the natural range from Senegal to Uganda were sampled and leaves of 118 individual trees were collected. An analysis of molecular diversity was carried out using random amplified polymorphic DNA (RAPD) markers. Fifteen random primers generated 67 polymorphic and 15 monomorphic RAPD loci ranging from size 1670 bp to 280 bp. Shannon's diversity index varied from Central Africa/Ndele (0.374) to Uganda/Amoya (0.350) but the differences between populations were smaller than the population standard errors. Correspondence analysis of unrooted neighbour-joining trees suggested that genetic distances between populations were correlated with geographic distances. This trend was confirmed by a Mantel test giving a coefficient of correlation between genetic and geographic distances of R = 0.88 (P = 0.0001). Result of AMOVA (analyses of molecular variance) showed that 14.8% (P = 0.002) of the RAPD variation was distributed among populations. Nested analysis of variance indicated that variance between the western and eastern groups of population represented 8.7% (P = 0.001) of the total variation and the variation amongst populations within group was 9.5% (P = 0.001). Eighty two percent of the variation was explained by variation amongst individuals within populations. The origin of genetic structure and level of diversity may be explained by the glacial refugia, the biological traits of Vitellaria paradoxa and by the impact of semi-domestication. Based on these results, sampling options of the natural populations are suggested for in or ex situ conservation. For the development of Vitellaria paradoxa breeding population, the sampling should consist of many individual trees selected within a few populations to capture a large proportion of variation.This revised version was published online in November 2005 with corrections to the Cover Date.     

云南河谷构树的遗传多样性研究

以云南省境内金沙江、元江、红河和怒江流域自然分布的构树为试材,采用AFLP分子标记技术对90份构树种质资源进行了遗传多样性分析研究.结果表明:筛选出的7对引物组合共扩增获得786条清晰可辩的条带,其中,多态性带632条,多态性条带百分率达80.4％,平均每对引物组合检测出90.3个多态位点.分布于4条水系流域的构树居群间,金沙江流域构树居群的遗传多样性水平最高,Nei's基因多样性指数为0.145 5,而元江流域构树居群的遗传多样性水平最低,Nei's基因多样性指数为0.1129.4个构树居群间的遗传分化系数为0.038 6,表明构树的遗传变异主要存在于居群内不同个体之间.在遗传距离为0.003时,4个构树居群可分为2组,第1组由金沙江流域的构树居群构成,第2组包含分布于红河、怒江和元江流域的3个构树居群.     

RAPD标记研究马褂木地理种群的遗传分化

利用RAPD标记对马褂木全分布区 15个种群的遗传多样性进行了分析。研究发现 ,马褂木具有丰富的遗传多样性 ,但东部种群因现有种群小 ,其遗传多样性明显地低于西部种群。种群遗传学研究揭示 ,由于小种群效应 ,以及缺乏有效的基因流和特有的致濒机制 ,马褂木种群间的遗传分化巨大 ,30 %以上的遗传变异存在于种群间。但在不同的地理区域 ,种群间的遗传分化程度不同。在西部分布区 ,地域上相距不远的种群其遗传距离较小 ,而在东部即使两个相邻的种群 ,其遗传距离也非常大。使用Nei’s遗传距离进行算术平均数的非加权成组配对法分析 (UPGMA) ,其聚类结果可以很好地将马褂木划分为西部和东部两个种源区 ,而西部种源又可再划分为西南亚区和华中亚区     

天然蒙古栎群体遗传多样性的RAPD分析

蒙古栎 (ＱｕｅｒｃｕｓｍｏｎｇｏｌｉｃａＦｉｓｃｈ)为壳斗科、栎属植物 ,主要分布区包括中国的华北、东北、内蒙古东部 ,朝鲜半岛 ,俄罗斯远东和蒙古的连续分布区及日本桦太、北海道的间断分布区 (吴晓春 ,1 993)。蒙古栎是我国东北的典型植被类型———红松阔叶林的主要伴生树种之一 ,同时在红松林被干扰破坏后是形成次生阔叶林面积较大的主要树种 ,也是分布区超过北纬 45°的唯一栎属植物。近年来 ,蒙古栎的分布区和群体数量呈扩张趋势 ,其原因除人为干扰外 ,可能涉及气候变化、物种特性、种内遗传多样性、种间互作等等。对蒙古栎群体…     

Effect of fluoride pollution on genetic variability of <Emphasis Type="Italic">Larix gmelinii</Emphasis> (Pinaceae) in East Siberia

The genetic variability of four natural populations of Larix gmelinii from East Siberia was determined and compared by RAPD analysis. Comparison of the RAPD profiles provided an estimation of variability in 193 RAPD fragments. More than 89% of these fragments were found to be polymorphic. The main genetic variability parameters of the two populations from Central Yakutia, a region free of fluoride pollution, had considerably higher values than those from East Transbaikalia, a region potentially affected by fluoride pollution (FLU, near a fluorite quarry growing on soils with a high natural content of fluorides). AMOVA revealed that 72.94% of the variation was within populations, while only 7.05% of the variation was between populations within geographical regions. The genetic diversity of the FLU fluoride-tolerant population was the lowest, but only slightly lower than that of a fluoride non-tolerant population from Chita, 50 km distant from FLU. Although this study demonstrates the absence of fundamental alterations of genetic structure within the populations of L. gmelinii growing on soils with a high content of fluorides, it is presumed that the reduction of genetic diversity was the genetic response of the FLU population to such an environmental stress as a constantly high concentration of fluorides within the soil.