Analysis of molecular genetic diversity and population structure in Amaranthus germplasm using SSR markers

We assessed the molecular genetic diversity and population structure of Amaranthus species accessions using 11 simple sequence repeat markers. A total of 122 alleles were detected, and the number of alleles per marker (N_A) ranged from 6 to 21 with an average of 11.1 alleles. The frequency of major alleles per locus ranged from 0.148 to 0.695, with an average value of 0.496 per marker. The overall polymorphic information content values were 0.436–0.898, with an average value of 0.657. The observed heterozygosity (H_O) and expected heterozygosity (H_E) ranged from 0.056 to 0.876 and from 0.480 to 0.907, with average values of 0.287 and 0.698, respectively. The average H_O (0.240) was lower than the H_E and gene flow (Nm), and showed substantial genetic variability among all populations of amaranth accessions. The sample groupings did not strictly follow the geographic affiliations of the accessions. A similar pattern was obtained using model-based structure analysis without grouping by species type. Knowledge of the genetic diversity and population structure of amaranth can be used to select representative genotypes and manage Amaranthus germplasm breeding programs.     

Plant genetic diversity and grazing management on the Qinghai-Tibetan Plateau: A case study of a dominant native wheatgrass (Elymus nutans)

Grazing management on the Qinghai-Tibetan Plateau in China concerns farmers, scientists and policy makers because of its geographical uniqueness. To investigate the effects of grazing on genetic diversity in native plants, the dominant perennial wheatgrass Elymus nutans was used as a model to examine genetic diversity and spatial population structure under different grazing intensities. Intra-population genetic diversity in three grazed areas was higher than un-grazed area; however, the genetic diversity decreased with increasing grazing intensity. Genotypes from four areas were separated into two groups (grazed and un-grazed) through a clustering analysis. Approximately 12% (G_st = 0.1487) of the DNA variation was apportioned among populations by AMOVA indicating a relatively low degree of inter-population genetic differentiation. Significant spatial auto-correlations and subpopulation structure were also observed within the four areas, and the genetic ‘patch’ size decreased with increasing grazing intensity. On the high and cold Qinghai-Tibetan Plateau, grazing is suitable for the maintenance of genetic diversity, and a light grazing intensity is optimal for the conservation of perennial grass germplasm and long-term grassland management.     

Examining the genetic diversity of Stipa grandis under various grazing pressures

The Stipa grandis steppe in the Inner Mongolia Autonomous Region occupies an area of 2798081 hm2. On the basis of the genetic variation, it was found that its adaptability to the environmental conditions under grazing pressure was significant. Using the Inter-Simple Sequence Repeat (ISSR) procedure, the changes to the genetic diversity of the Stipa grandis population under different grazing pressures were observed. Plant samples were collected from a series of grazing gradients of the Stipa grandis steppe in Dalinuoer National Nature Reserve in the Inner Mongolia (located at 116°38′–116°41′E and 43°25′–43°27′N.), which has the following vegetation types in abundance: Leymus chinensis is the constructive species; the dominant species include Stipa grandis, Cleistogenes squarrosa, and Artemisia frigida; the companion species is Potentilla acaulis and others. According to the grazing pressure, the following four grazing gradients were identified from the dwellings of the herdsmen to the enclosure site: (1) no grazing (CK enclosure site); (2) light grazing (LG); (3) moderate grazing (MG); (4) heavy grazing (HG). Young leaves of each Stipa grandis were collected during the growing season. The results showed that the Stipa grandis showed abundant genetic diversity despite the fact that certain polymorphic loci were lost; at the same time, new polymorphic loci emerged when grazing pressure increased; a total of 10 primers were used, and 74 bands were produced in total, of which 65 bands were polymorphic; the total percentage of polymorphism was 89%; the percentage of polymorphic loci of the Stipa grandis population decreased with the increase of grazing pressure; the percentage of polymorphic loci was 62.2% in the no-grazing (CK) population, 64.9% in the light-grazing (LG) population, 58.1% in the moderate-grazing (MG) population, and 56.8% in the heavy-grazing (HG) population; the genetic diversity of the population in the descending order using the Shannon's information index is as follows: (1) light grazing (0.3486); (2) no grazing (0.3339); (3) moderate grazing (0.3249); (4) heavy grazing (0.2735) with the same distributional pattern as the Nei's genetic diversity index. The test showed the following: As the grazing pressures increased, the change of genetic diversity decreased; the genetic differentiation coefficient among the population (Gst) was 0.1984, which showed the presence of small partial genetic diversity (19.8%) among populations; gene flow (Nm*) between primers varied from 0.9806 to 3.4463, and the mean gene flow (Nm*) was 2.0202; the UPGMA cluster figure that was constructed on the basis of the genetic distance matrix showed four populations that became genetically closer at each step: (1) The first group was the moderate-grazing (MG) population and the heavy- grazing (HG) population; (2) The second group consisted of the no-grazing (CK) population and the light-grazing (LG) population; (3) The two groups gathered together.     

Isozymes,plant population genetic structure and genetic conservation

Summary The exploration, conservation and use of the genetic resources of plants is a contemporary issue which requires a multidisciplinary approach. Here the role of population genetic data, particularly those derived from electrophoretic analysis of protein variation, is reviewed. Measures of the geographic structure of genetic variation are used to check on sampling theory. Current estimates justify the contention that alleles which have a highly localised distribution, yet are in high frequency in some neighbourhoods, represent a substantial fraction of the variation. This class, which is the most important class in the framing of sampling strategies, accounts for about 20–30% of variants found in 12 plant species. The importance of documenting possible coadapted complexes and gene-environment relationships is discussed. Furthermore, the genetic structure of natural populations of crop relatives might suggest the best structure to use in the breeding of crops for reduced vulnerability to pest and disease attack, or for adaptation to inferior environments. The studies reported to date show that whilst monomorphic natural populations do occur, particularly in inbreeding colonisers, or at the extreme margins of the distribution, polymorphism seems to be the more common mode. It is stressed here that the genetic resources of the wild relatives of crop plants should be systematically evaluated. These sources will supplement, and might even rival, the primitive land races in their effectiveness in breeding programmes. We may look forward to a wider application of gel electrophoresis in the evaluation of plant genetic resources because this technique is currently the best available for detecting genetic differences close to the DNA level on samples of reasonable size.     

蒙新区子午沙鼠种群的遗传多样性和遗传结构

为了探讨蒙新干旱区景观和环境因素对子午沙鼠种群遗传结构的影响,我们利用8 个多态性的微卫星位点,对子午沙鼠14 个局域种群的160 个个体进行了种群遗传多样性和遗传结构的分析。微卫星结果显示种群具有高的遗传多样性,微卫星位点的等位基因数(A)为22. 50 ±3.02 (19 ～ 28),多态信息含量(PIC)为0.912 ±0. 02 (0.872 ～ 0. 929);种群观测杂合度(HO )为0.68 ± 0. 19 (0. 52 ～0.85),期望杂合度(HE )为0.79 ± 0. 08(0. 71 ～ 0.85)。分子变异分析(AMOVA)结果显示,所得到的3 个组之间已发生显著的遗传分化(P < 0. 001)。
Mantel 检测结果表明,子午沙鼠种群遗传结构与地理距离无显著的相关性(P = 0.270 0),而在多元线性模型中,蒙新干旱区的海拔高度是影响种群遗传结构的关键因素。     

华木莲居群遗传结构与保护单元

华木莲(Sinomanglietia glauca)仅分布于江西宜春和湖南永顺, 是我国一级重点保护植物。前人采用RAPD、ISSR以及叶绿体SSCP(single-stranded conformation polymorphism)标记对华木莲进行了居群遗传学研究, 但未包括后发现的湖南居群或未检出居群内遗传变异。为了全面检测华木莲遗传多样性及其空间分布格局, 并据此确定保护单元, 本研究采用细胞核微卫星标记对华木莲所有4个居群共77个个体进行了居群遗传学分析。结果表明, 华木莲具有较低的遗传多样性(平均等位基因数A = 2.604, 平均期望杂合度H_E = 0.423)和较高的遗传分化(F_ST = 0.425)。STRUCTURE和主成分分析(Principal Coordinated Analysis, PCA)将4个居群首先分为江西、湖南两组, 江西的2个居群实际上是同一个自然繁育居群, 而湖南的2个居群则为2个分化明显的自然繁育居群。研究还发现湖南居群存在明显的杂合子过剩现象, 可能是小居群内随机因素造成的。研究结果表明华木莲可能在近期历史上遭受过强烈的瓶颈效应, 导致种群缩小、遗传多样性丧失和居群分化加剧, 需要加强对其进化潜力的保护。在制定保护措施时, 需要考虑其较高的遗传分化水平, 根据遗传结构可以将其划分为3个保护单元, 即湖南居群和江西居群分别为2个进化显著单元, 湖南居群进一步划分为2个管理单元(分别为朗溪乡云盘村和小溪乡鲁家村居群)。     

Population genetic structure of the Korean threadsail filefish (Stephanolepis cirrhifer) based on microsatellite marker analysis

The threadsail filefish Stephanolepis cirrhifer is one of the most important commercial fish resources in Korea. However, its natural populations have declined drastically over the last several decades. To investigate the genetic characteristics of the threadsail filefish for conservation and management purposes, its genetic variation was analyzed in Korean fishing grounds using 11 microsatellite loci. All populations showed moderate genetic diversity (mean number of alleles (N_A) = 8.6, expected heterozygosity (He) = 0.73), representing a slightly lower level of genetic variability than other congeneric species. The presence of a weak genetic population substructure was detected (F_ST = 0.023, R_ST = 0.030, P < 0.001), but this substructure did not feature significant isolation based on the distance between populations. This differentiation may be primarily attributable to genetic differences between populations from Geomundo and other localities, based on relative low level of gene flow, a high level of pairwise F_ST and R_ST or the position of this population in a phylogenetic tree. An analysis of molecular variance (AMOVA) also revealed a greater part of the variation within the population and genetic differentiation among the samples analyzed and identified two genetic clusters. The possibility of a recent genetic bottleneck was observed in some populations of S. cirrhifer. Given the prevailing ocean currents and the intensity of threadsail filefish harvesting activities in Korea, the possibility that human harvest and stock enhancement activities may have partially contributed to our detected genetic pattern cannot be excluded. Management strategies that take these findings into account might lower the risk of harmful genetic change in the species.     

Population genetic structure of banana corm weevil Cosmopolites sordidus (Germar) in India

The population genetic structure of Cosmopolites sordidus (Germar), an economically important pest of bananas, was studied using the sequence data of the internal transcribed rDNA (ITS1+ITS2) and the mitochondrial ‘COI-tRNA^Leu-COII’ region from seventy nine individuals collected from six sampling locations in India. The ITS data revealed 70% within population variation and non-significant genetic differentiation estimates suggesting lack of phylogeographic sub-structuring. 49% within population variation and highly significant genetic differentiation values were obtained with the mitochondrial data. The Mantel test revealed lack of correlation between genetic and geographic distance with both the markers. Demographic expansion of the populations was confirmed by the star shaped haplotype networks, demographic tests and mismatch distribution curves using both the markers. Molecular diversity indices show a high haplotype diversity (Hd) but low nucleotide diversity (π) suggesting that the populations are closely related. Considering the low self-dispersal ability of the weevils, these results suggest that the range expansion of this banana pest in India has taken place mainly through transport of infested corms and plant material resulting in the weevils forming localised populations which are not genetically distinct from each other. The high gene diversity (Hd) has enabled the weevils to adapt to varying environmental conditions which could explain the range expansion of this pest in India. The observed discrepancy in the genetic differentiation estimates using these two markers can be attributed to the evolutionary dynamics of the nuclear and mitochondrial genomes.     

基于核基因和叶绿体基因的台湾水韭遗传多样性和遗传结构的分析

利用核基因和叶绿体基因对台湾水韭（Isoetes taiwanensis Devol）2个居群20个样本的遗传多样性和遗传结构进行了分析,并对台湾水韭的保育提出了建议。核基因的检测结果显示,2个居群共存在18个单倍型,单倍型多样性为0.9842,核苷酸多样性为0.00215,居群间基因流N_m为4.26,居群间分化系数G_st值为0.05543;叶绿体DNA的检测结果显示,单倍型数目为6,单倍型多样性为0.6211,核苷酸多样性为0.00326。其中台北居群的单倍型多样性和核苷酸多样性均比金门居群高。核基因和叶绿体基因的AMOVA分析显示,居群内的遗传差异远高于居群间。台湾水韭的核苷酸多样性相对其他水韭属植物较低,可能与台湾水韭的染色体倍型以及狭域分布有关。居群结构的形成可能和基因流以及奠基者效应有关。宜采用原地和迁地保护的方法对居群进行保护。     

Population genetic structure of the medicinal plant Vitex rotundifolia in China: implications for its use and conservation

Vitexrotundifolia L.is an important plant species used in traditional Chinese medicine.For its efficient use and conservation,genetic diversity and clonal variation of V.rotundifolia populations in China were investigated using inter-simple sequence repeat markers.Fourteen natural populations were included to estimate genetic diversity,and a large population with 135 individuals was used to analyze clonal variation and fine-scale spatial genetic structure.The overall genetic diversity (GD) of V.rotundifolia populations in China was moderate (GD=0.190),with about 40% within-population variation.Across all populations surveyed,the average within-population diversity was moderate (P = 22.6%; GD = 0.086).A relatively high genetic differentiation (Gst=0.587)among populations was detected based on the analysis of molecular variance data.Such characteristics of V.rotundifolia are likely attributed to its sexual/asexual reproduction and limited gene flow.The genotypic diversity (D=0.992) was greater than the average values of a clonal plant,indicating its significant reproduction through seedlings.Spatial autocorrelation analysis showed a clear within-population structure with gene clusters of approximately 20 m.Genetic diversity patterns of V.rotundifolia in China provide a useful guide for its efficient use and conservation by selecting particular populations displaying greater variation that may contain required medicinal compounds,and by sampling individuals in a population at >20 m spatial intervals to avoid collecting individuals with identical or similar genotypes.     

大石鸡兰州亚种的mtDNA种群遗传结构、单倍型分布和遗传多样性

大石鸡(Alectorismagna)是中国西北部的特有种。我们测定了大石鸡兰州亚种(A.m.lanzhouensis)8个地理种群106个样本的线粒体DNA控制区5′端458bp序列,研究其种群遗传结构和遗传多样性。27个变异位点共确定25种单倍型,其中单倍型M2广泛分布,而许多单倍型为一些地方种群特有。单倍型分布沿着南北方向变化,存在明显的地理结构。8个种群中核苷酸多样性最高的是定西种群,0.0069,最低的是海原种群,0.0028;单倍型多样性最高的是武山种群,0.86,最低的是北道种群,0.52。北方种群比南方种群具有更高的遗传多样性。系统发生树和单倍型分布表明,大石鸡兰州亚种存在两个明显的分支。溯祖理论、更新世冰期和花粉支持兰州亚种起源于兰州盆地,这个盆地是其遗传多样性的中心。     

High genetic differentiation and low genetic diversity in Incarvillea younghusbandii, an endemic plant of Qinghai-Tibetan Plateau,revealed by AFLP markers

Incarvillea younghusbandii Sprague (Bignoniaceae) is a perennial herbaceous plant endemic to Qinghai-Tibetan Plateau. As a species of medical and horticulture importance, I. younghusbandii is threatened by over exploitation and habitat fragmentation. In this study, we analyze the genetic diversity and population structure of I. younghusbandii using amplified fragment length polymorphism (AFLP) markers. Our data reveal very low levels of genetic diversity in seven natural populations across Tibet. Specifically, at population level, the average Nei's genetic diversity index (H_E) and Shannon's diversity index (I) were 0.063 and 0.096, respectively. In contrast, high genetic differentiation among populations (Gst = 0.6238, Φ_ST = 0.614) is detected. The results of Neighbor-joining cluster, PCO, and STRUCTURE assignment reveal consistent pattern, suggesting seven well-defined genetic groups that are concordant with their geographical origins. The possible mechanisms and implications of these findings for conservation are discussed.     

Isozyme analysis of genetic variability and population structure of Lactuca L. germplasm

Isozymes were used to investigate the genetic variability, population structure, and relationships of Lactuca germplasm. The isozyme systems revealed 16 putative loci of a total of 31 alleles. Out of these 16 loci, 11 were polymorphic. The average values of expected heterozygosity (H_e), observed heterozygosity (H_o), mean number of alleles per locus (A) and effective number of alleles per locus (A_e) were 0.2227, 0.266, 1.3005 and 1.369, respectively. The average fixation indices were lower than zero for most of the accessions studied, indicating an excess of heterozygotes. Genetic differentiation among accessions (F_ST) exhibited that 51.3% of the isozyme variation was recorded among accessions, and 48.7% of the genetic variation resided within accessions. The average values of total heterozygosity (H_T) and intra-accessional genetic diversity (H_S) were 0.352 and 0.171, respectively. Moreover, the inter-accessional genetic diversity (D_ST) ranged from 0 to 0.424 with an average of 0.18. Cluster analysis revealed that L. sativa cultivars were distributed throughout different Lactuca species. Thereby, isozymes results confirms the hypothesis of the polyphyletic origin of L. sativa. This high level of genetic variation proved that isozymes are efficient for polymorphism analysis of Lactuca germplasm.     

Population structure and diversity in cultivated and wild Luffa species

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基于粪便DNA的青藏高原雪豹种群调查和遗传多样性分析

基于非损伤性取样,利用mtDNA和微卫星DNA遗传标记,对来自青藏高原三江源国家级自然保护区、羌塘国家级自然保护区和甘肃党河南山地区等地的277 份疑似雪豹粪便样品进行了物种鉴定、个体识别和遗传多样性研究。结果显示：在190份成功扩增的mtDNA cyt b基因片段中,确定有89份属于雪豹;微卫星分析确定其属于48个不同的个体。在48个雪豹mtDNA cyt b基因片段中,共检测出13个多态性位点,定义了9个单倍型,单倍型多态性为0.776,核苷酸多态性为 1.50%, 9个单倍型的遗传距离为 0.009-0.058。研究表明在研究区域均存在雪豹分布,并具有一定的遗传多态性,地理距离分布较远的羌塘国家级自然保护区和甘肃党河南山地区的雪豹样品具有一定的遗传差异。     

克氏针茅草原群落物种多样性与生物量关系对放牧强度的响应

放牧强度引起的草原植物群落物种多样性与地上生物量变化是近年来草地生态系统研究的热点问题。以内蒙古锡林郭勒克氏针茅草原为研究对象,探究植物群落结构特征、物种多样性与地上生物量之间相互关系及其对不同放牧强度的响应。结果表明：随着放牧强度的增加植物群落结构逐步向退化方向演替;植物群落高度逐渐降低（P<0.05）,密度逐渐增加（P<0.05）,盖度总体呈下降趋势（P<0.05）;植物群落和原有群落优势种地上生物量总体呈下降趋势（P<0.05）,而退化指示物种的地上生物量逐渐增加（P<0.05）;轻度、中度放牧条件下群落物种Margalef指数、Pielou指数、Simpson指数均显著高于重度放牧（P<0.05）;地上生物量与Shannon-Wiener指数、Margalef指数和Pielou指数呈正相关关系,而与Simpson指数呈负相关关系。综上所述,克氏针茅草原植物群落结构和功能在不同放牧强度下产生不同的响应,适度放牧有利于提高群落物种多样性与生物量。     

Population structure and genetic diversity of black redhorse (Moxostoma duquesnei) in a highly fragmented watershed

Dams have the potential to affect population size and connectivity, reduce genetic diversity, and increase genetic differences among isolated riverine fish populations. Previous research has reported adverse effects on the distribution and demographics of black redhorse (Moxostoma duquesnei), a threatened fish species in Canada. However, effects on genetic diversity and population structure are unknown. We used microsatellite DNA markers to assess the number of genetic populations in the Grand River (Ontario) and to test whether dams have resulted in a loss of genetic diversity and increased genetic differentiation among populations. Three hundred and seventy-seven individuals from eight Grand River sites were genotyped at eight microsatellite loci. Measures of genetic diversity were moderately high and not significantly different among populations; strong evidence of recent population bottlenecks was not detected. Pairwise F_ST and exact tests identified weak (global F_ST = 0.011) but statistically significant population structure, although little population structuring was detected using either genetic distances or an individual-based clustering method. Neither geographic distance nor the number of intervening dams were correlated with pairwise differences among populations. Tests for regional equilibrium indicate that Grand River populations were either in equilibrium between gene flow and genetic drift or that gene flow is more influential than drift. While studies on other species have identified strong dam-related effects on genetic diversity and population structure, this study suggests that barrier permeability, river fragment length and the ecological characteristics of affected species can counterbalance dam-related effects.     

Fine-scale genetic structure of life history stages in the food-deceptive orchid Orchis purpurea

In natural plant populations, fine-scale spatial genetic structure can result from limited gene flow, selection pressures or historical events, but the role of each factor is in general hard to discern. One way to investigate the origination of spatial genetic structure within a plant population consists of comparing spatial genetic structure among different life history stages. In this study, spatial genetic structure of the food-deceptive orchid Orchis purpurea was determined across life history stages in two populations that were regenerating after many years of population decline. Based on demographic analyses (2001-2004), we distinguished between recruits and adult plants. For both sites, there was no difference in the proportion of polymorphic loci and expected heterozygosity between life history stages. However, spatial autocorrelation analyses showed that spatial genetic structure increased in magnitude with life history stage. Weak or no spatial genetic structure was observed for recruits, whereas adult plants showed a pattern that is consistent with that found in other species with a predominantly outcrossing mating system. The observed differences between seedlings and adults are probably a consequence of changes in management of the two study sites and associated demographic changes in both populations. Our results illustrate that recurrent population crashes and recovery may strongly affect genetic diversity and fine-scale spatial genetic structure of plant populations.     

高寒草原优势种紫花针茅叶片解剖结构对青藏高原高寒干旱环境适应性分析

随着气候变暖, 高寒草原分布面积逐步增加, 高寒草原植物如何适应高寒干旱环境的研究还比较缺乏。该研究通过分析高寒草原优势种紫花针茅(Stipa purpurea)不同地理种群叶片解剖结构特征差异及其与气候因子的相关性, 阐明紫花针茅叶片适应高寒环境的策略, 为理解高寒植物对高寒干旱胁迫环境的适应机制提供科学依据。在青藏高原不同地理位置选择8个紫花针茅种群, 选择成熟健康叶片用卡诺氏固定液固定, 将固定好的叶片带回实验室进行石蜡切片和染色, 用显微镜观察叶片结构, 并用数码相机拍摄, 然后用软件Image-pro plus 6对叶片结构进行测量。结果显示: 紫花针茅叶片普遍具有较厚的角质层, 可减少水分散失和抵御较强的辐射; 不同地理种群紫花针茅叶片解剖结构在厚壁细胞厚度、叶片厚度、导管直径、主脉导管腔面积/主脉维管束面积和维管束面积/叶横切面积等特征上存在较大差异, 以适应不同区域的生境。Pearson相关性和聚类分析结果表明紫花针茅叶片解剖结构与气候因子密切相关; 主成分和冗余分析结果表明在干旱区域紫花针茅叶片解剖结构主要受到蒸发量的影响, 而在相对湿润区域紫花针茅叶片解剖结构主要受生长季降水量、湿润系数和年降水量/年蒸发量影响。综上所述, 紫花针茅通过增加厚壁细胞减少水分散失, 同时增加导管直径、主脉导管面积/主脉维管束面积和维管束面积/叶横切面积等输水组织面积适应高寒干旱气候。