秦岭山区美容杜鹃五个野生种群遗传多样性的ISSR分析

美容杜鹃是杜鹃花科杜鹃花属常绿观花植物,也是秦岭地区的特有属,在维持当地生态平衡方面发挥着重要作用,但目前资源正在遭受严重的破坏。该文通过ISSR分子标记技术对秦岭地区5个美容杜鹃野生种群进行遗传多样性分析,了解美容杜鹃不同种群的遗传分化,从而为美容杜鹃野生种质资源保护策略的制定提供理论依据。用8个ISSR引物对5个天然种群的90个单株进行扩增,共扩增出78条带(平均每条引物产生9.75条带),其中65条带是多态的,多态位点占总位点的百分率为83%。种群总的Nei’s基因多样性指数为0.3386,Shannon信息多态性指数为0.4972,表明美容杜鹃总的遗传多样性水平较高。种群多态位点百分率为82.71%~90.25%,Shannon信息多态性指数为0.4161~0.5867,Nei’s基因多样性指数为0.3044~0.4122,表明美容杜鹃不同种群遗传多样性水平差异较大。其中镇安木王种群和柞水牛背梁种群的遗传多样性水平较高。AMOVA分析表明种群内的遗传变异(91.22%)大于种群间的遗传变异(8.78%)。UPGMA聚类分析表明5个种群的遗传分化程度与地理距离没有相关性。因此建议尽可能地保护美容杜鹃所有的天然种群原生境条件,以最大限度保护其遗传变异。由于镇安和柞水种群具有较高的遗传多样性,因此建议优先对这2个种群实施就地保护和迁地保护。     

草河口林场红松人工林遗传多样性的ISSR分析

应用ISSR分子标记技术对本溪草河口林场的红松（Pinus koraiensis）人工林两个种群的60个个体进行遗传多样性分析。14个ISSR引物共检测到90个位点,其中多态位点65个,多态位点比率0.722 2。Nei指数统计结果表明,红松人工林的遗传多样性喜鹊沟种群（0.278 9）大于烈士墓种群（0.271 2）,Shannon指数统计结果与Nei指数相同。研究证实草河口林场红松人工林保存了较多的遗传多样性。     

古尔班通古特沙漠荒漠肉苁蓉遗传多样性分析

为了全面了解古尔班通古特沙漠荒漠肉苁蓉居群分布的遗传多样性特点,本研究通过ISSR分子标记技术,利用Nei和Shannon等多样性指数对古尔班通古特沙漠中5个居群166个个体的荒漠肉苁蓉遗传多样性、荒漠肉苁蓉种群和种内的遗传多样性进行分析。在供试材料中,8个引物共扩增出144个多态位点,多态位点百分率达100%,5个居群的多态位点百分率差异在46.53%～77.78%之间。在物种水平上,Nei基因多样度(h)为0.260 4,Shannon多样性指数(I)是0.411 0。遗传变异分析表明,物种水平的居群间遗传分化系数Gst为0.222 2,居群间的基因流Nm为1.750 7。研究显示古尔班通古特沙漠中荒漠肉苁蓉多态位点比例高,各居群基因交流较多,不同居群间遗传变异并不明显,这些对肉从蓉资源有效地保护和利用具有重要意义。     

珍稀濒危药用植物黄檗野生种群遗传多样性的AFLP分析

黄檗(Phellodendronamurense)是我国珍贵的药用和用材树种,现已被列为国家二级保护植物。为了研究其遗传多样性和遗传结构,以制定有针对性的保护策略,作者采用8对荧光引物,对来自10个野生种群共计129个黄檗个体进行了AFLP分析。共扩增出1,704条谱带,其中多态带1,581条。遗传分化系数Gst为0.3420,即有34.20%的遗传变异来自种群间。黄檗物种水平的遗传多样性高于种群水平:在物种水平上,多态位点百分率PPL=92.77%,有效等位基因数Ne=1.4636,Nei’s基因多样性指数H=0.2316,Shannon信息指数I=0.4275;在种群水平上,PPL=54.20%,Ne=1.2487,H=0.1524,I=0.2371。UPGMA聚类结果表明,黄檗野生种群可分为四类,并且来自同一地区的黄檗个体有聚为一类的趋势,说明其遗传变异可能与生境有关。建议在各种群内大量采样,实施迁地保护;在长白山、完达山、大兴安岭、小兴安岭、千山以及燕山山脉的云蒙山等地区,均匀布点划分区域进行就地保护,保护种群完整性,促进种群自我更新。     

应用ISSR-PCR分析蒙古栎种群的遗传多样性

本研究应用ISSR标记技术对东北地区的优势树种蒙古栎(Quercus mongolica)的25个种群遗传多样性进行了分析, 目的是为蒙古栎早期选择提供依据。从60条ISSR引物中筛选出10个特异性强、稳定性好的引物进行ISSR分析。共获得位点数71个, 其中多态位点数56个, 多态位点百分率为78.87%。PopGene分析结果表明: 种群的平均多态位点百分率为45.2%, Shannon表型多样性指数(I)平均值为0.25, 具有较高的遗传多样性, 种群间存在一定程度的基因流(N_m为1.3818)和遗传分化(Nei’s信息多样性指数平均值为0.1068, G_st平均值为0.2657), 种群内的基因多样度占总种群的73.43%, 种群间占26.57%, 表明蒙古栎种群的变异主要来源于种群内。结合聚类分析和地理变异规律把种群划分为两个大的种群组: 小兴安岭种群组和长白山种群组。以上结果可为栎属种质资源的保护和利用以及物种分化研究提供基础资料。     

利用AFLP分子标记探讨蜡梅种质资源的遗传多样性

利用AFLP分子标记技术,对中国蜡梅种质资源7个野生种群的遗传多样性进行了研究。利用筛选出的3对引物,共扩增出253条谱带,其中218条多态带,多态位点占86.17% ;种群间的基因分化系数为0.2906,说明蜡梅基因多样性主要存在于种群内;种群总的Nei s基因多样性指数为0.2933,Shannon信息多态性指数为0.4487,蜡梅总的遗传多样性水平较高。蜡梅不同种群遗传多样性水平差异较大,种群多态位点百分率在65.44% ～87.16%之间,Nei s基因多样性指数为0.1653 ～0.4012,Shannon信息多态性指数为0.3132 ～0.5603。神农架种群(SN)和保康种群(BK)的遗传多样性水平较高。用NTSYS2.01版软件对样品进行UPGMA聚类分析,结果7个种群并没有按地理距离进行聚类。最后提出要对各蜡梅野生群体采取相应的迁地和就地保护措施。     

地被植物鹅绒委陵菜的遗传多样性研究

采用RAPD方法对不同海拔和植被盖度下的鹅绒委陵菜（Potentilla anserinaL.）6个种群的遗传多样性水平进行了研究。结果表明,13条随机引物共扩增出132条带,多态性位点为117;鹅绒委陵菜物种水平的遗传多样性较高,且种群间分化明,多态位点比率（88.64%）、Nei’s基因多样性（0.3180）、Shannon信息多样性指数（0.4732）均远高于多数克隆植物。6个种群内的遗传多样性水平变化较大,多态位点在各种群中分布不均衡,种群间分化系数（GST）为47.7%,基因流较低（Nm=0.5482）,多态位点比率在31.06%～74.24%之间,Shannon信息指数在0.1711～0.3625之间,Nei基因多样性指数在0.1164～0.2425之间。多样性水平的变化与海拔没有明显的相关性,而与生境盖度呈显著正相关。从而推断在盖度低、资源丰富的环境中,该物种可能更倾向于克隆繁殖。     

枫香自然种群遗传多样性的ISSR分析

利用ISSR分子标记技术,对浙江省内枫香自然种群的遗传多样性进行分析。用10个引物对5个枫香种群共100个个体的样品DNA进行扩增,共测得135个位点,其中多态位点为118个,多态位点百分率(P)为87.41%,Shannon指数(I)为0.4646,Nei指数(h)为0.3122,表明枫香总体水平的遗传多样性较高。各种群的多态位点百分率平均为59.11%,Shannon指数平均为0.3660,Nei指数平均为0.2543。P、I、h均显示北山种群最高,天台山种群最低。AMOVA分子差异分析显示:85.49%的变异存在于种群内,14.51%存在于种群间,基因分化系数(Gst)为0.1856,种群间的遗传分化程度较低。种群间的基因流(Nm)为2.1944。5个种群间的平均遗传距离为0.1199。利用UPGMA法对5个种群进行聚类分析,结果分为两大类群:白云山、天台山、北山和安岱后4个种群组成一大类群;大明山种群单独为另一类群。     

蜡梅种质资源遗传多样性的ISSR分析

利用ISSR分子标记技术,对蜡梅种质资源7个野生种群和2个栽培种群的遗传多样性进行了研究。用11条引物,共扩增出124条谱带,其中110条多态带,多态位点占88.70％。用POPEGEN1.31版软件对数据进行分析,结果显示：种群总的Nei’s基因多样性指数为0.272 6,Shannon信息多态性指数为0.411 7,蜡梅总的遗传多样性水平较高。蜡梅不同种群遗传多样性水平差异较大,种群多态位点百分率在52.94％~90.00％之间,Nei’s基因多样性指数为0.143 4~0.378 2,Shannon信息多态性指数为0.232 2~0.546 6。神农架种群（SN）和保康种群（BK）的遗传多样性水平较高。种群间的基因分化系数为0.353 6,种群内的遗传变异大于种群间的遗传变异。用NTSYS2.01版软件对样品进行UPGMA聚类分析,结果9个种群并没有按地理距离进行聚类。种群间的地理距离和遗传距离之间没有显著的相关性(r＝0.437 1,P＝0.921 3)。     

四川六个厚朴种群遗传结构

利用ISSR技术对四川6个野生厚朴种群的遗传结构进行了分析,以评估野生厚朴资源的遗传现状。10条ISSR引物共测到114个位点,其中多态位点93个,多态位点百分率(PPB)为81.58%,Nei遗传多样性指数(H)和Shannon信息指数(I)分别为0.320和0.469,表明厚朴物种水平具有较高的遗传多样性;相比之下,种群水平遗传多样性则相对较低(PPB、H、I分别平均为48.25%、0.189和0.277)。分子变异分析(AMOVA)表明,种群内部和种群间均存在极显著遗传分化(P0.001),其中,36.82%的变异存在于种群间,63.18%存在于种群内,种群间的基因分化系数(GST)为40.42%。厚朴种群间的基因流(Nm)为0.737,平均遗传距离为0.211,算术加权平均数法(UPGMA)显示厚朴6个种群被分为3大类群。Structure分析表明,厚朴种群间遗传结构具有独立性特点。     

Patterns of Genetic Variation in Swertia przewalskii, an Endangered Endemic Species of the Qinghai-Tibet Plateau

Swertia przewalskii Pissjauk. (Gentianaceae) is a critically endangered and endemic plant of the Qinghai-Tibet Plateau in China. RAPD and ISSR analyses were carried out on a total of 63 individuals to assess the extent of genetic variation in the remaining three populations. Percentage of polymorphic bands was 94% (156 bands) for RAPD and 96% (222 bands) for ISSR. A pairwise distance measure calculated from the RAPD and ISSR data was used as input for analysis of molecular variance (AMOVA). AMOVA indicated that a high proportion of the total genetic variation (52% for RAPD and 56% for ISSR) was found among populations; pairwise Φ _ST comparisons showed that the three populations examined were significantly different (p < 0.001). Significant genetic differentiation was found based on different measures (AMOVA and Hickory θ_B) in S. przewalskii (0.52 on RAPD and 0.56 on ISSR; 0.46 on RAPD and 0.45 on ISSR). The differentiation of the populations corresponded to low average gene flow (0.28 based on RAPD and 0.31 based on ISSR), whereas genetic distance-based clustering and coalescent-based assignment analyses revealed significant genetic isolation among populations. Our results indicate that genetic diversity is independent of population size. We conclude that although sexual reproduction and gene flow between populations of S. przewalskii are very limited, they have preserved high levels of genetic diversity. The main factors responsible for the high level of difference among populations are the isolation and recent fragmentation under human disturbance.     

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。     

Model‐based conservation planning of the genetic diversity of Phellodendron amurense Rupr due to climate change

Climate change affects both habitat suitability and the genetic diversity of wild plants. Therefore, predicting and establishing the most effective and coherent conservation areas is essential for the conservation of genetic diversity in response to climate change. This is because genetic variance is a product not only of habitat suitability in conservation areas but also of efficient protection and management. Phellodendron amurense Rupr. is a tree species (family Rutaceae) that is endangered due to excessive and illegal harvesting for use in Chinese medicine. Here, we test a general computational method for the prediction of priority conservation areas (PCAs) by measuring the genetic diversity of P. amurense across the entirety of northeast China using a single strand repeat analysis of twenty microsatellite markers. Using computational modeling, we evaluated the geographical distribution of the species, both now and in different future climate change scenarios. Different populations were analyzed according to genetic diversity, and PCAs were identified using a spatial conservation prioritization framework. These conservation areas were optimized to account for the geographical distribution of P. amurense both now and in the future, to effectively promote gene flow, and to have a long period of validity. In situ and ex situ conservation, strategies for vulnerable populations were proposed. Three populations with low genetic diversity are predicted to be negatively affected by climate change, making conservation of genetic diversity challenging due to decreasing habitat suitability. Habitat suitability was important for the assessment of genetic variability in existing nature reserves, which were found to be much smaller than the proposed PCAs. Finally, a simple set of conservation measures was established through modeling. This combined molecular and computational ecology approach provides a framework for planning the protection of species endangered by climate change.     

A Study of Conservation Genetics in Cupressus chengiana,an Endangered Endemic of China,Using ISSR Markers

ISSR markers were used to analyze the genetic diversity and genetic structure of eight natural populations of Cupressus chengiana in China. ISSR analysis using 10 primers was carried out on 92 different samples. At the species level, 136 polymorphic loci were detected. The percentage of polymorphic bands (PPB) was 99%. Genetic diversity (H _e) was 0.3120, effective number of alleles (A _e) was 1.5236, and Shannon’s information index (I) was 0.4740. At the population level, PPB = 48%, A _e=1.2774, H _e=0.1631, and I=0.2452. Genetic differentiation (G _st) detected by Nei’s genetic diversity analysis suggested 48% occurred among populations. The partitioning of molecular variance by AMOVA analysis indicated significant genetic differentiation within populations (54%) and among populations (46%; P < 0.0003). The average number of individuals exchanged between populations per generation (N _m ) was 0.5436. Samples from the same population clustered in the same population-specific cluster, and two groups of Sichuan and Gansu populations were distinguishable. A significantly positive correlation between genetic and geographic distance was detected (r=0.6701). Human impacts were considered one of the main factors to cause the rarity of C. chengiana, and conservation strategies are suggested based on the genetic characters and field investigation, e.g., protection of wild populations, reestablishment of germplasm bank, and reintroduction of more genetic diversity.     

Inter-simple sequence repeat (ISSR) diversity within Monarda fistulosa var. brevis (Lamiaceae) and divergence between var. brevis and var. fistulosa in West Virginia

Inter-simple sequence repeat (ISSR) banding patterns were used to examine genetic diversity within and among populations ofMonarda fistulosa var.brevis, a rare taxon restricted to several populations in limestone glades and barrens in eastern West Virginia and Virginia. More than 34% of the total ISSR diversity in var.brevis occurred among populations, which is high when compared to the few other rare species that have been examined for ISSR variation. Prior studies demonstrated that var.brevis is morphologically distinct from the more widespread var.fistulosa, and that the differences are maintained when the two varieties are grown together in a uniform environment. The present study utilizing ISSR markers indicated that the two varieties are distinct, though quite similar genetically, and this is concordant with prior investigations documenting their morphological and habitat differences. However, the ISSR results suggest that the two varieties have diverged relatively recently and/or there is a low level of gene flow between them.     

The impact of genetic diversity on the accuracy of DNA barcoding to identify species: A study on the genus Phellodendron

DNA barcoding is widely used in species identification, but there is considerable controversy regarding the extent of sampling in research methods. Some scholars have proposed that this small sample size underestimates the intraspecific genetic diversity, which would impact on the accuracy of DNA barcoding to identify species. In study, we selected all Phellodendron species (including P. amurense Rupr., P. chinense Schneid., and P. chinense var. glabriusculum Schneid.) as the materials, collected 59 P. amurense samples from 35 populations greatly to represent the genetic diversity, and analyzed the haplotype, genetic distance, barcoding gap, and Neighbor‐Joining (NJ) trees based on psbA‐trnH and internal transcribed spacer gene sequences. Additionally, a sampling simulation was conducted to assess the correlation between genetic diversity and the number of populations. Finally, analysis of critical geographical populations was performed. Based on analysis of haplotype, genetic distance, barcoding gap, and NJ trees, we found that eight P. amurense samples impacted on the effectiveness of DNA barcoding, which genetic information were very important to identify Phellodendron species. Moreover, the result of the NJ tree analysis performed the small‐scale P. amurense sample size did not completely match the objective phylogenetic relationship in Phellodendron. In simulation sampling analysis, the data showed the genetic diversity indexes at the same population level gradually decreased and stabilized as the number of simulation sampling populations increased. We found that 1–2 samples from over 24 populations based on uniform geographical distribution could represent 80% of the genetic diversity of P. amurense and ensure authenticity and reliability of DNA barcoding. Thus, we proposed it is particularly important adequately samples to cover infraspecific genetic diversity in order to ensure identification accuracy of DNA barcoding.     

Assessing genetic diversity of Elymus sibiricus (Poaceae: Triticeae) populations from Qinghai-Tibet Plateau by ISSR markers

Inter-simple sequence repeats (ISSR) markers were used to assess the genetic diversity and population structure in eight populations of Elymus sibiricus L. from the southeast of Qinghai-Tibet Plateau of China. Of the 100 primers screened, 13 produced highly reproducible ISSR bands. Using these primers, 193 discernible DNA fragments were generated with 149 (77.2%) being polymorphic, indicating considerable genetic variation at the species level. In contrast, there were relatively low levels of polymorphism at the population level with the percentage of polymorphic bands (PPB) ranging from 44.04 to 54.92%. The mean gene diversity (H_E) was estimated to be 0.181 within populations (range 0.164–0.200), and 0.274 at the species level. A high level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (33.1%), Shannon's index analysis (34.5%), Bayesian method (33.2%) and AMOVA analysis (42.5%). No significant statistical differences (analysis of molecular variance [AMOVA], P = 0.08) in ISSR variation were found between the sample collection regions. However, among populations (42.5% of the variance) and within populations (57.5% of the variance), there were significant differences (P < 0.001). Populations shared high levels of genetic identity. This pattern of genetic variation was different from that for most of inbreeding Triticeae species reported. In addition, a geographical pattern of population differentiation, where the populations from south and north of sampling sites were clearly separated from each other, was revealed by both the cluster and principal coordinates analyses. Generally, the result of this study indicates that E. sibiricus contains high molecular variation in its populations. The implications of these results for the conservation of the species are discussed.     

Genetic structure analysis of natural <Emphasis Type="Italic">Sargassum muticum</Emphasis> (Fucales,Phaeophyta) populations using RAPD and ISSR markers

Sargassum muticum is important in maintaining the structure and function of littoral ecosystems, and is used in aquaculture and alginate production, however, little is known about its population genetic attributes. In this study, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to investigate the genetic structure of four populations of S. muticum and one outgroup of S. fusiforme (Harv.) Setchell from Shandong peninsula of China. The selected 24 RAPD primers and 19 ISSR primers amplified 164 loci and 122 loci, respectively. Estimates of genetic diversity with different indicators (P%, percentage of polymorphic loci; H, the expected heterozygosity; I, Shannon’s information index) revealed low or moderate level of genetic variations within each S. muticum population, and a high level of genetic differentiations were determined with pairwise unbiased genetic distance (D) and fixation index (F _ST ) among the populations. The Mantel test showed that two types of matrices of D and F _ST were highly correlated whether from RAPD (r = 0.9706, P = 0.009) or ISSR data (r = 0.9161, P = 0.009). Analysis of molecular variance (AMOVA) was conducted to apportion the variations among and within the S. muticum populations. It indicated that variations among populations were higher than those within populations, being 55.82% verse 44.18% by RAPD and 55.21% verse 44.79% by ISSR, respectively. Furthermore, the Mantel test suggested that genetic differentiations among populations were related to the geographical distances (r > 0.6), namely, conformed to the IBD (isolation by distance) model, as expected from UPGMA (unweighted pair group method with arithmetic averages) cluster analysis. On the whole, the high genetic structuring among the four S. muticum populations along the distant locations was clearly indicated in RAPD and ISSR analyses (r > 0.9, P < 0.05) in our study.     

珍稀濒危植物太行菊遗传多样性的ISSR分析

利用ISSR分子标记技术对太行山特有濒危物种太行菊11个自然居群的遗传多样性进行研究。用10个引物对11个居群的122个样品进行扩增,共得到150个扩增位点,其中多态性位点149个,多态位点百分率(PPL)为99.33%。POPGENE分析显示,太行菊具有较高的遗传多样性(H=0.2149,I=0.3455)。沁阳市大西天居群的遗传多样性水平最高(H=0.1910,I=0.2969),山西陵川县大双村居群的遗传多样性水平最低(H=0.1356,I=0.2155)。Nei’s遗传多样性分析表明,11个自然居群间出现了较高的遗传分化(基因分化系数Gst=0.2566,基因流Nm=1.4488)。生境的的片段化和基因流障碍可能是导致太行菊居群间遗传分化显著的主要原因。通过对太行菊居群遗传多样性和遗传结构的分析,该文提出了一些保护策略。