Reproductive biology and population genetic structure of Kandelia candel (Rhizophoraceae), a viviparous mangrove species

The pollination biology, mating system, and population genetic structure of Kandelia candel were investigated. Field observations on its pollination and reproductive biology suggested that this species is pollinator dependent for fruit set, and bee activities can lead to substantial geitonogamous selfing. Quantitative analysis of the mating system parameters was performed using progeny arrays assayed for allozyme markers. Multilocus outcrossing rates (t(m)) were estimated to be 0.697 ± 0.091 and 0.797 ± 0.062 in two populations. In comparison to other plant species with mixed-mating system, the level of allozyme variation was very low in the 13 populations sampled along the coastlines of Hong Kong. At the species level, the proportion of polymorphic loci was 20%, number of alleles per locus was 1.2, and heterozygosity was 0.0362. The total gene diversity was primarily distributed within populations (H(S )= 0.0339), and the coefficient of genetic differentiation among populations was low (G(ST )= 0.064). This pattern of population genetic structure suggests that gene flow, primarily in the form of water-dispersed seedlings in viviparous mangrove species, is not as limited as previously thought. However, microgeographic pattern in allele frequency at the marker loci could still be detected between the western and eastern coastal populations.     

Development of microsatellite markers in a mangrove tree species Aegiceras corniculatum (Myrsinaceae)

Aegiceras corniculatum is an ecologically important mangrove tree species. We isolated 18 polymorphic microsatellite loci from this species. These loci provided microsatellite markers with polymorphism of two to eight alleles per locus. The observed and expected heterozygosities ranged from 0.050 to 0.550, and from 0.097 to 0.736, respectively. These markers will contribute to research on the conservation, genetic diversity and mating patterns of A. corniculatum.     

中国卵叶海桑遗传多样性的ISSR研究

卵叶海桑 (Sonneratiaovata)是海桑科濒危红树植物 ,在我国仅分布于海南文昌清澜自然保护区内。采用简单序列重复区间扩增 (ISSR)分子标记技术对该天然居群和东寨港红树林自然保护区引种的人工居群共 3个居群 3 9个个体进行了遗传变异分析。 1 1个引物共扩增出 1 85条带 ,其中 1 2 7条具多态性 ,多态位点百分率为 68.65 %。在居群水平上相对较低 ,多态位点百分率 3 6.76%～ 5 4.5 9% ,平均值为 47.2 1 %。Nei的基因多样性、Shannon信息指数在物种水平上分别为 0 .1 41 1和 0 .2 2 92 ;在居群水平上平均值分别为 0 .1 2 0 9和0 .1 91 0。Nei的遗传分化系数Gst表明 :87.5 8%遗传变异分布在居群内 ,1 2 .42 %的遗传变异分布在居群间。居群间的遗传一致度达 0 .970 7。东寨港迁地保护的人工居群有效地保护了卵叶海桑的遗传多样性。     

Clonal diversity in the rare Calamagrostis porteri ssp. insperata (Poaceae): comparative results for allozymes and random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) markers

Four populations of the rare, highly clonal grass Calamagrostis porteri ssp. insperata were examined using allozymes and the two polymerase chain reaction (PCR)-based markers, random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) bands. Only one of the 15 allozyme loci was variable and two alleles were detected, both of which were found in two populations, while only one genotype was detected in the other two populations. ISSR and RAPD markers detected more genotypes within populations than did allozymes. ISSR markers detected more diversity than RAPD markers in three of the four populations examined. In one population, no RAPD diversity was found whereas eight different genotypes were found among the 10 plants with ISSR markers. This diversity is present despite rare flowering, no documented occurrence of seed set in natural populations and very low seed set with experimental pollinations, all of which suggest that sexual reproduction rarely occurs. The subspecies is self-compatible, but seed initiation is lower in selfed ovules; also, there is high embryo abortion regardless of pollen source. Variation detected by RAPD and ISSR primers may reflect higher levels of sexual reproduction in the past, very rare sexual reproduction in extant populations, somatic mutations, or a combination of the three. Although the PCR-based markers identify several multilocus genotypes within populations, it is not known whether these all represent distinct genets generated by sexual reproduction or result from somatic mutations in the old, perennial and highly clonal plants.     

Assessing hybridization in natural populations of Penstemon (Scrophulariaceae) using hypervariable intersimple sequence repeat (ISSR) bands

Inferences regarding hybridization rely on genetic markers to differentiate parental taxa from one another. Intersimple sequence repeat (ISSR) markers are based on single-primer PCR reactions where the primer sequence is derived from di- and trinucleotide repeats. These markers have successfully been used to assay genetic variability among cultivated plants, but have not yet been tested in natural populations. We used genetic markers generated from eight ISSR primers to examine patterns of hybridization and purported examples of hybrid speciation in Penstemon (Scrophulariaceae) in a hybrid complex involving P. centranthifolius , P. grinnellii , P. spectabilis and P. clevelandii . This hybrid complex has previously been studied using three molecular data sets (allozymes, and restriction-site variation of nuclear rDNA and chloroplast DNA). These studies revealed patterns of introgression involving P. centranthifolius , but were unsuccessful in determining whether gene flow occurs among the other species, and support for hypotheses of diploid hybrid speciation was also lacking. In this study, we were able to fingerprint each DNA accession sampled with one to three ISSR primers and most accessions could be identified with a single primer. We found population- and species-specific markers for each taxon surveyed. Our results: (i) do not support the hybrid origin of P. spectabilis ; (ii) do support the hypothesis that P. clevelandii is a diploid hybrid species derived from P. centranthifolius and P. spectabilis ; and (iii) demonstrate that pollen-mediated gene flow via hummingbird vectors is prevalent in the hybrid complex.     

广西红树植物桐花树种群遗传多样性分析

采用RAPD-PCR方法探讨广西3个不同生境下桐花树种群的遗传多样性和遗传分化。结果表明:3个不同生境桐花树RAPD扩增多态百分率为20.2%,3个不同生境桐花树种群两两之间的遗传距离分别为0.195、0.169、0.26,平均遗传距离为0.208。同一种群不同个体的扩增多态百分率最高为37.28%,其次为20.93%,最小的为19.32%。Shannon’s遗传多样性指数3个种群分别为0.331、0.225和0.17,其大小顺序与多态百分率的结果一致。种群内遗传多样性比率为62.3%,种群间遗传多样性比率为37.7%。说明广西3个不同生境的桐花树种群的遗传变异大部分存在种群内,种群间遗传变异较小。     

Mating system and genetic diversity of a rare desert legume Ammopiptanthus nanus (Leguminosae)

Ammopiptanthus nanus is an endangered evergreen shrub endemic to the deserts of central Asia and plays an important role in delaying further desertification. We examined allozyme variation and AFLP diversity in A. Nanus populations and investigated the mating system of this species using progeny arrays assayed for poly-morphic allozyme loci. Mating system analysis in the Keyi'eryongke'er population showed low levels of out-crossing, and strong inbreeding depression. Low levels of genetic variation were detected at both population (allozyme, Pp=14.0%,A=1.14, He=0.031; AFLP, Pp=14.5%, Shannon's information index I=0.063) and species (allozyme, Pp=21.1%,A=1.21, He=0.040; AFLP, Pp=20.9%, I=0.083) levels; while moderate genetic differentia-tion existed among populations, as indicated by allozymes (GST=0.081) and AFLP (GST=0.151-0.193). Founder effect, bottlenecks in evolutionary history, the mixed mating system and co-ancestry may have influenced the level of genetic diversity in A. Nanus. Markers of both types provide new insights for conservation management, indicating that the Biao'ertuokuoyi and Keyi'eryongke'er populations should be given priority for in situ conser-vation and regarded as seed sources for ex situ conservation.     

Assessing the genetic diversity of tobacco germplasm using intersimple sequence repeat and inter-retrotransposon amplification polymorphism markers

The genetic diversity of 118 tobacco accessions, including flue‐cured tobacco, sun‐/air‐cured tobacco, burley tobacco, oriental tobacco and wild tobacco, was characterised using intersimple sequence repeat (ISSR) and inter‐retrotransposon amplification polymorphism (IRAP) markers. ISSR and IRAP banding patterns and genetic distance (GD) values showed the low level of genetic diversity within and among cultivated tobacco types. There was higher GD and average heterozygosity among wild tobacco types than those among cultivated tobacco. Genetic diversity of tobacco germplasm was low, with a high level of genetic identity (>0.77) between the different types. However, neighbour‐joining cluster analysis of marker‐based GDs showed that the accessions from the same tobacco type, as classified by manufacturing quality traits, were nearly clustered into the same group. These results will help in the formulation of appropriate strategies for variety improvement in tobacco, and ISSR and IRAP markers of the genetic diversity will contribute to further study and improvement of tobacco.     

Analysis of the genetic structure of Rhodiola rosea (Crassulaceae) using inter-simple sequence repeat (ISSR) polymorphisms

The genetic diversity and differentiation of eleven R. rosea populations from different parts of its wide area of occurrence were studied by ISSR markers. Using eight primers, 252 DNA fragments were generated, and 243 of those DNA fragments were found to be polymorphic, indicating a high genetic variability at the species level (P = 96.4%, h = 0.176, SI = 0.291). Relatively low levels of diversity were determined at the population level (P 30.6-59.1%, h 0.088-0.165, SI 0.137-0.257). AMOVA analysis revealed that the majority of the genetic variation was within populations (65.42%), and the variance among populations was 34.58%. Cluster analysis revealed two groups of R. rosea populations; these groups likely represent distinct evolutionary lines in the species, which are different in genetic structure, evolutionary history and chorological migration routes.     

Analysis of genetic relationship in mutant silkworm strains of Bombyx mori using inter simple sequence repeat （ISSR） markers

Amplified inter simple sequence repeats (ISSR) markers were used to determine genetic relationships among mutant silkworm strains of Bombyx mori. Fifteen ISSR primers containing simple sequence repeat (SSR) motifs were used in this study. A total of 113 markers were produced among 20 mutant swains, of which 73.45% were found to be polymorphic. In selected mutant genetic stocks, the average number of observed allele was (1.7080±0.4567), effective alleles (1.5194±0.3950) and genetic diversity (Ht) (0.2901±0.0415). The dendrogram produced using the unweighted pair group method with arithmetic means (UPGMA) and cluster analysis made using Nei's genetic distance resulted in the formation of one major group containing 6 groups separated 20 mutant silkworm strains. Therefore, ISSR amplification is a valuable method for determining the genetic variability among mutant silkworm swains. This efficient molecular marker would be useful for characterizing a considerable number of silkworm swains maintained at the germplasm center.     

甘肃省冬虫夏草遗传多样性的ISSR分析

对采自甘肃省3个冬虫夏草主产区域6个具有代表性地方的18个样本进行ISSR分析。15条ISSR引物共扩增得到条带清晰并呈多态性的95条谱带,每一引物扩增获得的ISSR条带数在3－9条之间,扩增片段集中在300－3,000bp。基于遗传相似性系数（GS）、不加权成对群算术平均法（UPGMA）构建的系统树分析,可以看出：分离自同一株虫草上不同部位的样本无显著遗传差异;同一地点样本间的遗传分化较小;不同地域的样本间存在着较大的遗传分化,遗传多样性较丰富。同时6个地方的冬虫夏草明显地分为3个区域,而在同一区域不同     

Assessment of genetic diversity in Salvadora persica L. based on inter simple sequence repeat (ISSR) genetic marker

Studies on the genetic variation in marginal populations and differentiation between them are essential for assessment of best gene conservation strategies and sampling schemes. In this study, ISSR markers were used to establish the level of genetic relationships and polymorphism 50 genotypes of Salvadora persica collected from 6 different regions of Hormozgan province. The ISSR analysis with 9 anchored primers also generated 105 scorable loci, of which 85 were polymorphic (80.95%). Parameters of genetic diversity and its partitioning were calculated. The genetic analysis demonstrated that S. persica maintain relatively high genetic diversity (PIC was 0.63, Na was 1.27 and Ho and He were 0.15 and 0.17 respectively). The coefficient of genetic differentiation among populations based on FST equaled 0.20. Genetic identities between population's pairs were high (mean I?=?0.88). These values are high as compared with other widespread congener species. Cluster analysis based on the Unweighted Pair Group Method with Arithmetic Averages (UPGMA) revealed 3 main clusters for the ISSR data. The levels of genetic diversity maintained within populations of S. persica indicate that an appropriate sampling design for ex situ safeguarding should capture the majority of genetic diversity found within these taxa to help ensure the long term viability of this species. Furthermore, it could be inferred that ISSR markers are suitable tools for the evaluation of genetic diversity and relationships within the Salvadora persica.     

等位酶分析的遗传学基础

在等位酶分析中,对酶谱的正确解释是获取遗传学资料的基础,酶谱作为酶基因的表现型,是由该种酶蛋白质的四级结构情况（亚基的数目）、在亚细胞分室中的分布（位点的数目）以及所分析样品的倍性和基因型的情况所决定的。术语“酶型”被建议用来记录和描述各种情况下的酶谱的不同。在进行生物多样性和分子系统学研究中,如果直接把酶谱上带的数目的多少作为遗传多样性大小的指标,或把带的多少及迁移率的大小作为数量性状进行聚类和分枝分析将会得出非常错误的结论。     

新疆地区番茄潜叶蛾遗传多样性的ISSR分析

[目的]番茄潜叶蛾已成为世界性番茄的重要害虫,对番茄产业的发展造成了严重的威胁,研究其遗传多样性有利于揭示不同地理种群的遗传变异结构。[方法]采用ISSR分子标记技术分析了20个地理种群番茄潜叶蛾的遗传多样性和遗传结构特征。[结果]15条引物扩增出137条ISSR条带,其中多态性条带占96.35%,所有个体显示了各自独特的ISSR图谱。ISSR的标记遗传多样性结果表明,20个番茄潜叶蛾地理种群遗传距离大小范围为0.0065~0.1623,遗传一致度范围为0.8502~0.9935。种群变异来源分析表明,25.36%的遗传变异来自种群间,74.64%的变异来源于种群内部。UPGMA系统发育分析结果表明,各地理种群的聚类与地理位置无较强的关联性。[结论]番茄潜叶蛾尚处在入侵早期阶段,且具备频繁入侵和多点的特征。防控上要注意加强检疫,阻绝多点入侵来源。     

闽江流域野生蕉(Musa itinerans)遗传多样性和遗传结构的ISSR分析

采用ISSR分子标记技术对福建省闽江流域10个野生蕉自然居群的遗传多样性和遗传结构进行分析,结果显示:12条ISSR引物共检测出117个条带,105个多态性条带,多态性百分率为89.7%;野生蕉Nei遗传多样性指数为0.244,Shannon's信息指数为0.381,其中三明野生蕉遗传多样性水平最高,且不同自然居群间遗传多样性指数之间具有显著性差异;总遗传变异系数为0.589,基因流为0.349,居群间的遗传分化程度高于居群内;基于遗传距离的聚类结果与模型聚类结果均聚为3大类,分别为沙溪支流的三明野生蕉类群、闽江上游及附近支流的南平野生蕉类群和闽江下游的福州野生蕉类群。研究认为,闽江流域野生蕉资源丰富的遗传多样性主要来源于自然居群间生境异质化所引起的高频率遗传变异,且三明野生蕉类群的遗传多样性和遗传分化程度最高,可能是福建野生蕉的起源中心,也是野生蕉资源开发和利用的最主要群落。此外,水流是闽江流域野生蕉遗传迁移最关键的自然主导因素。     

等位酶分析的遗传学基础（续）

在等位酶分析中,对酶谱的正确解释是获取遗传学资料的基础,酶谱作为酶基因的表现型,是由该种酶蛋白质的四级结构情况（亚基的数目）、在亚细胞分室中的分布（位点的数目）以及所分析样品的倍性和基因型的情况所决定的。术语“酶型”被建议用来记录和描述各种情况下酶谱的不同。在进行分子系统学研究中,如果直接把酶谱上带的数目的多少作为遗传多样性大小的指标,或把带的多少及迁移率的大小作为数量性状进行聚类和分枝分析将会得出非常错误的结论。     

Genetic diversity in Arabidopsis thaliana L. Heynh. investigated by cleaved amplified polymorphic sequence (CAPS) and inter-simple sequence repeat (ISSR) markers

In this study, we investigated genetic diversity among 37 accessions in Arabidopsis thaliana from Eurasia, North Africa and North America using morphological traits and two polymerase chain reaction (PCR)-based marker systems: cleaved amplified polymorphic sequences (CAPS) and inter-simple sequence repeats (ISSR). Cluster analysis based on genetic similarities calculated from CAPS data grouped the accessions roughly according to their geographical origin: one large group contained accessions from Western, Northern and Southern Europe as well as North Africa, a second group consisted of Eastern European and Asian continental accessions. North American accessions were interspersed into these groups. Contrary to the CAPS analysis, the dendrogram obtained from the ISSR data did not reflect the geographical origin of the accessions, and the calculated genetic distances did not match the CAPS results. This could be attributable to an uneven genomic distribution of ISSR markers as substantiated by a database search for ISSR binding sites in A. thaliana genomic DNA sequence files, or to the ISSR's different mode of evolution. We recommend CAPS markers for diversity analysis in A. thaliana because a careful selection of markers can ascertain an even representation of the entire genome.     

Intersimple sequence repeat (ISSR) polymorphisms as a genetic marker system in cotton

We studied the applicability of intersimple sequence repeat (ISSR) polymorphism in cotton. We found that: (i) the resolving power of agarose gels is poor relative to that provided by sequencing gels; (ii) fluorescent labelling of ISSR amplification primers produced numerous scorable bands; (iii) primer mixing (double priming) generated more bands than the sum of fragments resulting from two single primers, although an unexplained disappearance of several larger fragments also reproducibly occurred; (iv) ISSR fingerprinting patterns are highly heritable; and (v) double priming ISSR is an easy and informative genetic marker system in cotton for revealing both inter‐ and intraspecific variations.     

不同龙眼资源遗传多样性的SCoT和ISSR 比较分析

应用SCoT和ISSR标记对36份龙眼资源和1份近缘种龙荔的遗传多样性进行分析。结果表明:12对SCoT引物共扩增出127条带,平均每条引物扩增10.58条带;15条ISSR引物共扩增出117个条带,平均扩增7.8条带。UPGMA聚类结果表明:SCoT标记和ISSR标记分别在相似系数0.672和0.685水平上,均可将37份材料分成6大类群,SCoT和ISSR标记均适用于龙眼材料的遗传多样性分析,如果将两种标记的数据进行综合分析,可以缩小单一标记的误差。研究结果为龙眼种质资源的保存和利用提供了重要的依据。     

海南岛红树植物海桑遗传多样性的ISSR分析

海桑 (Sonneratia caseolaris)是海桑科红树植物 ,在我国仅天然分布于海南万宁、琼海、文昌等地。采用 ISSR分子标记技术对所有天然种群和海南东寨港红树林自然保护区引种的人工种群共 4个种群 86个个体进行了遗传变异分析。 11个引物共扩增出 2 39条带 ,其中 194条具多态性 ,多态位点百分率为 81.17%。在种群水平上多态位点百分率 4 0 .5 9%～ 5 0 .2 1% ,平均值为4 5 .71%。 Nei的基因多样性、Shannon信息指数在物种水平上分别为 0 .2 10 0和 0 .32 5 6 ,在种群水平上平均值分别为 0 .14 6 8和0 .2 2 10。 Nei的遗传分化系数 Gst和 AMOVA分析表明种群间已发生了较高的遗传分化。种群间的遗传一致度为 0 .90 11。估测的种群间的基因流为 0 .5 787。依据 Nei的遗传距离对不同种群进行 U PGMA聚类 ,聚类结果为横山种群 (HS)和东寨港种群(DZG)聚在一起 ,万宁种群 (WN)和琼海种群 (QH)聚为一类。Mantel检验表明遗传距离与地理距离之间有一定的正相关 ,但不显著。种群遗传多样性与环境因子间的相关性分析表明 :海桑种群遗传多样性水平与各环境因子间相关性均不显著。因东寨港引种种群的遗传多样性明显低于天然种群 ,为保护遗传多样性 ,应加强对琼海、万宁种群的就地保护和迁地保护工作。