Genetic diversity and population structure of Taxus cuspidata in the Changbai Mountains assessed by chloroplast DNA sequences and microsatellite markers

Taxus cuspidata, a tree species with high economic value because of its anticancer properties, is experiencing severe reduction in populations across its range in China. We examined one chloroplast DNA (cpDNA) region (petA-psbE) and 9 nuclear simple sequence repeats (SSRs) loci variations among seven populations in the Changbai Mountains of China to investigate the levels of genetic diversity and population structure. A moderate level of haplotype diversity (H_T = 0.625), low nuclear microsatellite diversity (H_E = 0.261 ± 0.028), significant genetic differences (F_ST = 0.065) and substantial gene flow (Nm = 2.806) were observed. Most of the total genetic variation was partitioned within the population (87.8% and 94.0% for cpDNA and SSRs, respectively). Our haplotype identification permutation tests revealed that G_ST > N_ST, indicating an absence of phylogeographic structure in T. cuspidata. Neither STRUCTURE nor UPGMA analyses showed any geographic pattern in T. cuspidata populations. By comparatively analyzing the genetic diversity and survival situation of T. cuspidata, our results provide a theoretical foundation for the resource protection, utilization cultivation and breeding of this valuable plant.     

箭叶淫羊藿居群形态及遗传多样性比较研究

箭叶淫羊藿（Epimedium sagittatum）是淫羊藿属中分布最广、形态变异最大、分类学最难处理的一个物种,不同箭叶淫羊藿居群其形态、活性成分等差异较大,质量极其不稳定。本实验选择湖北罗田等12个不同的箭叶淫羊藿地理居群,在武汉植物园进行同园栽培,分析其主要形态数量性状及遗传多样性。结果发现箭叶淫羊藿不同居群在形态上表现出各自明显的差异。基于AFLP数据进行的遗传多样性分析显示,各居群聚类关系与地理分布密切相关,柳州居群（LZ）、西南居群（CL、HH、ZY）、华东华中居群（YT、WN、NF、HS、QZ、SG、LY、LT）依次分出。而形态分层聚类分析显示各居群形态变异复杂,只有部分与遗传多样性有一定的相关性。本研究结果对箭叶淫羊藿分类研究及资源筛选具有重要指导作用。     

Population genetic structure of Bactrocera dorsalis based on cox1 sequences from Bangladesh and neighboring countries

Oriental fruit fly, Bactrocera dorsalis (Hendel), is a destructive and highly polyphagous invasive fruit fly species of numerous fruit crops in global agriculture. Population genetic structure of this species from five different locations of Bangladesh was examined with other samples (collected from GenBank) from 15 sites of neighboring Asian countries. A fragment of 770 bp mitochondrial DNA cox1 was used to investigate the genetic diversity and the relationship between genetic patterns and geographical distribution of B. dorsalis. A total of 232 variable sites (33.23% of the 698 bp aligned consensus sequences) and 419 unique haplotypes were identified from 710 individuals. Indices of genetic diversity suggested that without exclusion from geographical areas, B. dorsalis retained a relatively high degree of genetic diversity. A demographic assessment [Tajimas’ D test, Fu’s Fs test and sum of square deviation (SSD values)] revealed that both current and historical variables performed a significant role in deciding the weak genetic structure with some exceptions. In Bangladesh, high levels of genetic diversity with a weak genetic structure indicated that the severity of this pest might increase in the future. Proper management techniques should be taken to overcome the future severity of this kind of destructive insect.     

Genetic divergence in nuclear genomes between populations of <Emphasis Type=Italic>Fagus crenata</Emphasis> along the Japan Sea and Pacific sides of Japan

Genetic diversity and structure in Fagus crenata were studied by analyzing 14 nuclear microsatellite loci in 23 populations distributed throughout the species’ range. Although population differentiation was very low (F _ST = 0.027; R _ST = 0.041), both neighbor-joining tree and Bayesian clustering analyses provided clear evidence of genetic divergence between populations along the Japan Sea (Japan Sea lineage) and Pacific (Pacific lineage) sides of Japan, indicating that physical barriers to migration and gene flow, notably the mountain ranges separating the populations along the Japan Sea and Pacific sides, have promoted genetic divergence between these populations. The two lineages of the nuclear genome are generally consistent with those of the chloroplast genome detected in a previous study, with several discrepancies between the two genomes. Within-population genetic diversity was generally very high (average H _E = 0.839), but decreased in a clinal fashion from southwest to northeast, largely among populations of the Japan Sea lineage. This geographical gradient may have resulted from the late-glacial and postglacial recolonization to the northeast, which led to a loss of within-population genetic diversity due to cumulative founder effects.     

利用AFLP分析西南特色猕猴桃的遗传多样性

为分析西南地区特色猕猴桃(Actinidia Lindl.)的遗传多样性,建立并优化了猕猴桃DNA 多态性分析的AFLP 体系,AFLP 标记体系为300 ng 基因组DNA 用EcoR Ⅰ/Mse Ⅰ (15 U/5 U)于37℃下酶切2 h,加接头后的混合物稀释5 倍用于预扩增,预扩增产物再稀释10 倍后进行选择性扩增.结果表明,筛选的22 对引物进行扩增反应,共得到979 条条带,其中多态性条带为649 条.采用优化的AFLP 体系,对西南地区的10 种猕猴桃50 个样本进行了遗传多样性分析,包括普通品种中华猕猴桃、美味猕猴桃,及特色品种硬毛猕猴桃、毛被猕猴桃、革叶猕猴桃、四萼猕猴桃、狗枣猕猴桃、葛枣猕猴桃、京梨猕猴桃和紫果猕猴,结果表明种内和种间的聚类关系明显;斑果组(sect. Maculatae)和净果组(sect. Leiocarpae)的种间有聚类交叉,并呈现地理区域性分布.     

中国壳斗科植物空间多样性格局研究

采用地理信息系统技术,制作空间分布图、从空间上计算多样性格局指数,研究中国壳斗科植物属、种的空间多样性分布格局。结果显示,云南南部、广西北部和广东北部的属、种数量均较多,是中国壳斗科植物多样性的重要分布地区,甘肃南部、陕西南部、河南西部及南部是壳斗科植物向南、向北扩散的重要通道;从多样性指数来看,种的多样性指数值均比属的值高,但均匀度指数却是属的值高;当属或种的数量为1时,其所占面积、占景观的比例、斑块数量、最大斑块指数、景观形状指数均最大,随着属或种的数量逐渐增加,其多样性明显提高,但其各项指标基本呈依次降低的趋势。通过对壳斗科植物空间多样性格局进行量化研究,获取了中国壳斗科植物空间多样性分布规律及多样性格局数量特点,利用地理信息系统技术可以使多样性研究体现出空间性和定量化的特征。     

Population genetics of threatened wild plants in Japan

Approximately one-fourth of Japan's native plant species are threatened with extinction. To conserve these species, it is critical to evaluate genetic diversity at species-level and population-level. Some factors, including population size and geographic distribution, are known to influence the population genetic diversity of wild plant species. This article briefly reviews the population genetic studies that have been conducted on wild threatened plants in Japan. A large population size or wide geographic distribution does not always lead to large genetic diversity, suggesting that historical factors such as speciation processes and population expansion often play more important roles in determining genetic diversity than the number of remnant individuals. The mating system of a species also affects genetic diversity; predominantly selfing species tend to have smaller genetic diversity than outcrossing congeners. Another issue of concern in the conservation genetics of wild plants in Japan is the genetic diversity of insular endemics, because Japan consists of many islands, and the insular flora contains many endemic and threatened species. Previous studies on endemic plants on the Bonin and the Ryukyu Islands are reviewed. Compared to the cases of the Bonin Islands or other oceanic islands, there is much larger genetic diversity in plants endemic to the Ryukyu Islands. This difference is probably the result of the differences in the geological history of these islands. Electronic Publication     

Genetic structure of Himalayan marmot (Marmota himalayana) population alongside the Qinghai–Tibet Railway

In order to verify the effect of social behavior and geographical isolation on the genetic structure of the Himalayan marmot (Marmota himalayana) population, we examined the genetic diversity of Himalayan marmots alongside the Qinghai–Tibet Railway using microsatellite markers. Eight microsatellite loci were used to examine 120 animals of 4 populations: Ulan (U), Delhi (D), Tuotuohe (T) and Ando (A). The results show that: (1) Himalayan marmots alongside the Qinghai–Tibet Railway are highly genetically diversified. The allele number (Na), effective allele number (Ne), observed heterozygosity (Ho), Nei’s expected heterozygosity (He) and polymorphism information content (PIC) of the total Himalayan marmot population were 4.75, 3.0332, 0.6990, 0.6672, 0.6102, respectively. (2) Himalayan marmots may be able to avoid inbreeding by a mechanism that will prevent the genetic diversity reduction caused by their social lifestyle. Heterozygote excess was observed at most loci. The inbreeding coefficients within the subpopulation (F_IS), in the total population (F_IT), the differentiation index of population (F_ST), and the gene flow (Nm) were ?0.2265, ?0.0477, 0.1458, and 1.4646, respectively. (3) The genetic differentiation of the Himalayan marmot population was in accordance with Wright’s “isolation by distance” theory. The Mantel test indicates that the correlation between genetic distance and geographic distance was significant (P < 0.05, r = 0.698). (4) Each of the four geographical populations had moderate differentiation. Both geographic distance and isolation could affect the population genetic structure of the Himalayan marmot. The maximum gene flow (3.5915), the smallest genetic differentiation index (0.0651), the lowest genetic distance (0.0700) and the highest genetic identity (0.9526) were all between the Ulan population and Delhi populations. (5) The cluster analysis, based on Nei’s standard genetic distance, showed that the populations of Delhi and Ulan were first merged in a cluster, and then Tuotuohe population was merged in the clustering. The Ando population was the last element in the clustering.     

Genetic structure of Himalayan marmot (Marmota himalayana) population alongside the Qinghai–Tibet Railway

In order to verify the effect of social behavior and geographical isolation on the genetic structure of the Himalayan marmot (Marmota himalayana) population, we examined the genetic diversity of Himalayan marmots alongside the Qinghai–Tibet Railway using microsatellite markers. Eight microsatellite loci were used to examine 120 animals of 4 populations: Ulan (U), Delhi (D), Tuotuohe (T) and Ando (A). The results show that: (1) Himalayan marmots alongside the Qinghai–Tibet Railway are highly genetically diversified. The allele number (Na), effective allele number (Ne), observed heterozygosity (Ho), Nei’s expected heterozygosity (He) and polymorphism information content (PIC) of the total Himalayan marmot population were 4.75, 3.0332, 0.6990, 0.6672, 0.6102, respectively. (2) Himalayan marmots may be able to avoid inbreeding by a mechanism that will prevent the genetic diversity reduction caused by their social lifestyle. Heterozygote excess was observed at most loci. The inbreeding coefficients within the subpopulation (F_IS), in the total population (F_IT), the differentiation index of population (F_ST), and the gene flow (Nm) were ?0.2265, ?0.0477, 0.1458, and 1.4646, respectively. (3) The genetic differentiation of the Himalayan marmot population was in accordance with Wright’s “isolation by distance” theory. The Mantel test indicates that the correlation between genetic distance and geographic distance was significant (P < 0.05, r = 0.698). (4) Each of the four geographical populations had moderate differentiation. Both geographic distance and isolation could affect the population genetic structure of the Himalayan marmot. The maximum gene flow (3.5915), the smallest genetic differentiation index (0.0651), the lowest genetic distance (0.0700) and the highest genetic identity (0.9526) were all between the Ulan population and Delhi populations. (5) The cluster analysis, based on Nei’s standard genetic distance, showed that the populations of Delhi and Ulan were first merged in a cluster, and then Tuotuohe population was merged in the clustering. The Ando population was the last element in the clustering.     

中国大鲵四种群的遗传结构和地理分化

为了确定栖息地的破碎化和片断化引起中国大鲵的地理分化和遗传结构变异,本文测定了来自广西、河南、陕西和湖南4个地理种群的28条大鲵的mtDNAD loop基因全序列。根据这4个地理种群的地理分布,分成珠江单元(广西种群)、黄河单元(河南种群)和长江单元(湖南和陕西种群)。通过ClustalX、MEGA2.0、DnaSP4.0、Arlequin1.1分析发现,全序列长度为771bp,其中64个多态性核苷酸变异位点,占全部碱基数的8.26%。转换和颠换分别为6和2个,插入/缺失11个。27个单倍型间的序列差异平均为1.32%。3个单元的单倍型多样性指数和核苷酸多样性指数值都偏低,而且珠江单元的这两个指数值都低于长江和黄河两个单元。珠江单元和黄河、长江单元之间分化程度显著(P<0.001),而长江和黄河单元之间差异不显著(P>0.05)。地理单元内分化程度占99.31%,而单元间只有0.69%,表明遗传差异主要发生在单元内,而且各地理单元之间的基因流较频繁。构建的NJ树和MP树显示,27个单倍型呈现一种混杂的分布格局,并未分成代表3个地理单元的聚合群。     

缢蛏种群遗传多样性的周年变异分析

利用线粒体COI标记分析了福建省宁德市漳湾镇横屿村滩涂5个月份缢蛏群体(S3,S5,S7,S9,S11)的遗传结构,以期评估不同月份时期种群的遗传多样性差异。基于线粒体COI基因的结果表明,5个缢蛏群体的平均核苷酸差异数位于2.7836-3.6894之间,核苷酸多样性指数位于0.0050-0.0066之间,遗传多样性水平大致表现为S3和S5群体较高于S7和S9群体,明显高于S11群体。AMOVA分析结果显示,群体间遗传变异量占总变异的7.18%,而群体内变异达到了92.82%,说明遗传差异主要来自于群体内部。由此可见,从3月份到11月份,缢蛏群体的遗传多样性水平呈现出下降趋势,尤其是在11月份,差异最为明显。     

Natural hybridization within seed sources of shortleaf pine (Pinus echinata Mill.) and loblolly pine (Pinus taeda L.)

Shortleaf and loblolly pine trees (n = 93 and 102, respectively) from 22 seed sources of the Southwide Southern Pine Seed Source Study plantings or equivalent origin were evaluated for amplified fragment length polymorphism (AFLP) variation. These sampled trees represent shortleaf pine and loblolly pine, as they existed across their native geographic ranges before intensive forest management. Using 17 primer pairs, a total of 96 AFLPs between shortleaf pine and loblolly pine were produced and scored on the sample trees and two control-pollinated F1 interspecies hybrids and their parents. In addition, the well known isocitrate dehydrogenase (IDH) isozyme marker was scored for all trees. IDH detected two putative hybrids among the loblolly pine samples and two among the shortleaf pine samples, while either 13 or 12 putative hybrids were detected using all AFLP markers and IDH and either NewHybrids or Structure software, respectively. Results of this study show that later generation hybrids can be reliably identified using AFLP markers and confirmed that IDH is not a definitive marker for detecting hybrids; that is, at least in some seed sources, the alternative species’ IDH allele resides in the source species. Based on all the markers, hybridization frequency varied geographically, ranging from 30% in an Arkansas seed source to 0% in several other seed sources. The hybridization level was higher in populations west of the Mississippi River than in populations east of the river; the shortleaf pine hybridization rates were 16.3% and 2.4% and the loblolly pine rates were 4.5% and 3.3%, west and east of the river, respectively. The results suggest that hybridization between these two species is significant but varies by seed source and species, and the potential for the unintended creation of hybrids should be considered in forest management decisions regarding both natural and artificial regeneration.     

Genetic variation of Cordyceps sinensis, a fruit-body-producing entomopathogenic species from different geographical regions in China

Cordyceps sinensis is one of the most valuable medicinal fungi in the Orient. It is naturally distributed in the eastern extension area of the Qinghai-Tibet plateau, at an altitude over 4000 m high. In order to investigate genetic variation and evolutionary relationships of C. sinensis from different geographical regions, 17 isolates of C. sinensis were collected from different provinces and the complete sequences of rDNA ITS were determined. On the basis of 5.8S rDNA and ITS region analysis, it was clearly shown that the ITS sequences within C. sinensis are highly homologous regardless of geographical origin. The distance values between the sequences in this study were lower than 0.03. This implied that C. sinensis from different geographic regions are the same species; they are not different species or a species complex. The results also showed that distance values between C. sinensis and Hirsutella sinensis are of the same order as those within C. sinensis from different geographic regions. This confirmed our previous results that C. sinensis should only have H. sinensis as its asexual stage whatever the geographic region from which the samples were collected. An rDNA ITS clone library was established to obtain further evidence for the interpretation of the fungal community structure from C. sinensis and to confirm the accuracy of the taxonomic identities produced by directly sequencing the rDNA ITS region. The discrimination between intraspecies of C. sinensis might provide additional data for the authentication of medicinal material at the species or intraspecies level.     

Genetic diversity analysis of Rhodothermus reflects geographical origin of the isolates

The genetic diversity and relationships of 81 Rhodothermus isolates from different geothermal environments in Iceland were examined by analysis of electrophoretically demonstrable allelic variation of 13 genes encoding enzymes. All the enzymes were polymorphic. A total of 71 distinctive multilocus genotypes (electrophoretic types, ETs) were identified. The mean genetic diversity per locus (H _t ) was 0.586. The relatively high genetic variance observed within Rhodothermus isolates from different locations is most likely the result of genetic changes occurring independently in the locations studied. A high G _st value (0.284) indicates that a considerable part of the variance observed is due to differences between locations. Cluster analysis revealed two major groups of ET clusters diverging at a genetic distance of 0.75, reflecting strongly the geographic origin of isolates. Estimation of the association index (I _A) indicates that Rhodothermus marinus is a clonal species in which recombination events occur rarely. Partial or whole sequencing of the 16S rRNA genes of Rhodothermus isolates grouping at genetic distance of 0.40 confirmed that all the isolates belonged to the species Rhodothermus marinus. The results of this study confirm that, despite phylogenetic and phenotypic similarity, genetic diversity within Rhodothermus marinus is quite high. Received: January 21, 2000 / Accepted: April 27, 2000