宁波地区野生换锦花遗传多样性的ISSR分析

为了解宁波地区野生换锦花的生存状况及对目前环境的适应能力,本研究采用ISSR分子标记对6个野生居群的遗传多样性水平和遗传结构进行分析。结果表明,10条ISSR引物共扩增出74个条带,其中多态性条带67个,多态性位点百分比(PPL)为90.54%。宁波地区野生换锦花在物种水平上的遗传多样性较高,PPL为90.54%,Nei’s基因多样性(He)为0.2700,Shannon’s指数(Ho)为0.4144;但在居群水平上遗传多样性较低,PPL平均值为76.35%,He平均值为0.2517,Ho平均值为0.3800。基因分化系数(Gst)为0.0679,基因流(Nm)为6.8654。AMOVA分子变异分析结果表明,在总的遗传变异中,有95.85%的变异发生在居群内,仅有4.15%的变异发生在居群间。UPGMA聚类结果显示6个居群间的亲缘关系较近,其可分为两大类群,其中北仑、慈溪和象山3个居群组成一类;奉化、镇海和舟山3个居群组成一类。Mantel检验的结果表明,居群间的地理距离和遗传距离之间无显著相关性。本研究结果为宁波地区野生换锦花种质资源的保护及合理开发利用提供了理论依据。     

华东竹黄菌不同居群遗传分化的RAPD分析

为了解竹黄地理居群间的遗传分化,本研究采用随机引物扩增多态性DNA分子标记技术对江苏、安徽和浙江3省的8个居群共32个竹黄样本进行了遗传多样性分析。从50个RAPD引物中筛选得到了5个随机引物,对供试材料的DNA进行扩增,共检测出77个位点,其中多态性位点52个,多态性位点比率为67.53%。UPGMA聚类分析结果表明,这8个居群分为三类：安吉居群、临安居群、宜兴居群、广德居群和泾县居群聚为一类;宁国居群和休宁居群聚为一类;淳安居群单独为一类。遗传多样性分析表明8个竹黄居群中,淳安居群的遗传多样性水平最高,安吉居群的遗传多样性水平最低。Nei＇s基因多样性指数和Shannon信息指数均表明竹黄物种水平的遗传多样性高于居群水平。     

泰国产方斑东风螺养殖群体遗传多样性的RAPD分析

用随机扩增多态性DNA(RAPD)技术对泰国产方斑东风螺养殖群体的遗传多样性进行检测,从100个随机引物中筛选出21个引物对方斑东风螺的DNA进行扩增,结果表明:21个引物共检测到222条清晰且重复性好的条带,每个引物可扩增出4~16条带,分子量在200~2200bp之间,其中多态位点为156个,占70.27%;群体的Shannon多样性指数为0.2818,Nei基因多样性指数为0.2491;个体间最大遗传距离为0.291,最小遗传距离为0.066。通过与其他贝类遗传多样性的研究结果比较,可初步判断泰国产方斑东风螺养殖群体的遗传多样性比较丰富。     

2个中国藏獒群体遗传多样性及遗传分化的研究

本研究以青海土种犬和藏狮犬为对照,采用RAPD技术对河曲藏獒和青海藏獒群体的遗传多样性及遗传分化进行了分析。研究结果表明,河曲藏獒和青海藏獒群体的多态性位点百分率分别为85.53%和98.16%,平均多态性位点百分率为91.85%。藏獒群体遗传变异分析显示两个藏獒群体的有效等位基因数(Ne)、Nei氏平均预期基因杂合度(H)和Shannon遗传信息指数(I)的平均值分别为1.5354、0.3191和0.4807;且两个藏獒群体93.09%的变异来自群体内,仅6.91%变异来自群体间;两个群体间的基因流为3.3679。研究还发现两个藏獒群体之间的Nei氏标准遗传距离(D)为0.0505。本研究结果说明藏獒群体内遗传变异丰富,不同地域的藏獒群体之间存在广泛的基因交流,群体之间遗传分化程度很低,这为藏獒品种资源保护与合理开发利用提供了参考。     

青藏高原野生老芒麦种质醇溶蛋白遗传多样性分析

采用酸性聚丙烯酰胺凝胶电泳(A-PAGE) 对采集自青藏高原的54份野生老芒麦（Elymus sibiricus L.）种质进行遗传多样性分析,获得下述结果:（1）供试材料共分离出42条带纹,多态率达92.86%。4个电泳分区（α、β、γ、ω）的平均Shannon指数为0.4627,Nei-Li遗传相似系数（GS）变异范围为0.2424~0.9767,平均值为0.5822。说明供试野生老芒麦材料具有较为丰富的醇溶蛋白遗传多样性。（2）对所有材料的聚类分析和主成分分析发现,在GS值为0.562的水平上供试材料可聚成4个大类,绝大部分来自于相同或相似生态地理环境的材料聚成一类,主成分分析显示了相似的结果,即醇溶蛋白图谱类型与材料的生态地理环境具有一定的相关性。（3）基于Shannon 多样性指数估算了5个老芒麦地理类群内和类群间的遗传分化,发现类群内遗传变异占总变异的68.17%,而类群间的遗传变异占总变异的31.83 %。（4）对各地理类群基于Nei氏无偏估计的遗传一致度的聚类分析表明,各地理类群间的遗传分化与其所处的地理生态环境具有较高的相关性。     

紫茎泽兰群体遗传多样性及遗传结构的AFLP分析

紫茎泽兰是我国最为严重的入侵物种之一。本研究以中国境内的紫茎泽兰为材料，用AFLP方法研究了其群体遗传多样性及遗传分化。用筛选出的3对荧光引物，对5个地区62个群体进行AFLP分析，共扩增出490条带，其中多态性带328条，群体的遗传多样性较高(P = 66.9%，H = 0.171，I = 0.268)，主要存在于群体内，群体间的遗传分化系数Fst为0.287。AMOVA分析表明，在地区水平紫茎泽兰的遗传分化主要存地区内，群体内遗传分化占 70.25%，群体间遗传分化占21.71%，地区间的遗传分化仅占8.04%。UPGMA聚类分析结果表明，62个地理群体主要分为四大类群，并且有明显的地缘关系。Sigmaplot分析表明海拔是影响紫茎泽兰遗传多样性主要地理因子。Mantel检验表明遗传距离与地理距离成正相关(r = 0.34)，但也有例外。由此推断风媒传播可能是紫茎泽兰的主要传播方式，水媒传播可能是紫茎泽兰的另一主要传播途径；随着紫茎泽向东、向北扩散，新入侵地区的遗传多样性逐步降低。     

细鳞鱼三个野生种群的遗传多样性AFLP分析

细鳞鱼是我国一种珍贵的淡水经济鱼类,在人类和环境因素的影响下数量急剧下降。本文采用扩增片段长度多态性（Amplified Fragment Length Polymorphism, AFLP）技术对牡丹江（Mudanjiang River, MD）、鸭绿江（Yalujiang River, YL）和乌苏里江（Wusuli River, WSL）的三个细鳞鱼野生种群共72个个体进行了遗传多样性分析。结果显示,12对选择性扩增引物共扩增得到559个位点,其中多态性位点541个,多态性百分比为96.78％。文中对3个群体的Shannon多样性指数,Nei氏基因多样性等参数进行了分析。其总基因多样性Ht平均值为0.3512±0.0208;种群内基因多样性Hs平均值为0.2137±0.0152;群体间的基因多样性Dst为0.1375。基因分化系数Gst为0.3914,种群内的基因多样性占总群体的60.85%,种群间为39.15%,而基因流系数Nm 为0.7776。分子方差分析（AMOVA）表明：群体平均遗传分化系数Fst为0.55336,变异来源有44.84%来自群体间,55.16%来自群体内。三个群体中牡丹江群体的细鳞鱼种内多态性比例最高,而乌苏里江群体最低。     

中国主要棉花栽培群体的遗传多样性分析

利用ISSR分子标记对我国棉花栽培品种中的38个陆地棉(Gossypium hirsutum)和6个海岛棉(Gossypium bar-badense)共44个品种(按来源划分为6个品种群,6个海岛棉品种均属于新疆品种群体)进行遗传多样性分析表明,筛选出的11个引物在所有品种中共扩增出90条带,其中70条为多态性条带,多态性条带百分率(PPB)达77.78%,Popgene32软件分析结果表明,新疆品种群的遗传多样性水平最高[PPB=91.43%,H(基因多样性指数)=0.3769,I(信息指数)=0.5464],其次是中棉所品种群,河南品种群排位第三,而河北、山东和山西3个品种群体的遗传多样性水平相差较小,其中山西品种群遗传多样性水平最低(PPB=58.57%,H=0.2484,J=0.3587).6个地方棉花栽培群体在总的遗传变异中有82.42%存在于群体内,群体间的遗传变异仅占总变异的17.58%,群体间分化系数Gst=0.1759,群体间总的基因流的估算值.Nn=2.34.聚类分析结果表明,山西、山东和河北品种群体的亲缘关系最近,先聚合在一起,然后依次与河南和中棉所品种群体聚类,最后与亲缘关系最远的新疆品种群体聚合.     

湖南典型茶树地理种群遗传多样性

利用RAPD分子标记技术,对湖南典型茶树安化云台山种(Camellia sinensis var.sinensis)、城步峒茶(C.sinensis var.assamica cv.Duntsa)、汝城白毛茶(C.ptilophylla)和江华苦茶(C.sinensis var.assamica cv.Jianghua)4个地理种群的240个单株的遗传多样性、种群内和种群间的遗传变异进行了研究。结果表明,21个10碱基随机引物共检测到226条谱带,其中多态性谱带为201条,占88.9%。遗传多样性分析结果显示:Shannon's多样性指数为0.43,种群内变异占74.7%,而种群间变异占25.3%;Nei's指数群体总基因多样性(HT)为0.37,群体内平均基因多样性(HS)为0.28,群体间的基因多样性(HST)为0.09,群体Nei's基因分化系数(GST)为0.23,说明76.7%的变异存在于种群间,群体内的变异占了总变异的23.3%,与Shannon's多样性指数相比基本一致,均表明种群内有较丰富的遗传变异;种群间的基因流(Nm)为0.74,显示种群间的基因交流有限。     

利用SSR和RAPD标记分析中国部分地区晚疫病菌群体遗传多样性

利用SSR和RAPD对来自福建、黑龙江、河北和内蒙古4个地理群体的80个马铃薯晚疫病菌（phytophthora infestans）株进行了遗传多样性分析。13对SSR引物共扩增出76条谱带,多态性条带比率78.9%,相似系数变化范围0.00～0.42之间;筛选出的14条RAPD引物共扩增出189条谱带,多态性条带比率95.2%,相似系数变化范围0.04～0.66之间。遗传多样性分析表明,在4个群体中,福建群体的多样性更为丰富。遗传相似性分析显示,黑龙江和内蒙古两个群体间的遗传相似性最高,而福建和河北两个群体间的遗传相似性最低。聚类分析显示,来自南方福建的菌株与来自北方黑龙江、河北和内蒙古的菌株亲缘关系较远,且福建群体分布于更多的聚类组,显示出更高的遗传变异度。     

Conservation genetics and implications for restoration of Hemigenia exilis (Lamiaceae), a serpentine endemic from Western Australia

Hemigenia exilis (Lamiaceae) is a rare plant endemic to serpentine soils of the Goldfields of Western Australia. The species was presumed extinct until 1995, when it was re-discovered on a nickel ore deposit. To delineate the origin and extent of seed collection for rehabilitation after mining, and to identify the impact of removing one population due to mining, we assessed level and partitioning of genetic variation and differentiation. Twelve populations were sampled for DNA fingerprinting using the random amplified polymorphic DNA (RAPD) technique. Ten primers produced 89 bands, 97% being polymorphic. Genetic diversity within populations ranged from 0.197 to 0.409, averaging 0.38 at the species level, which is high compared with most other endemic species. Heterozygosity within populations ranged from 0.355 to 0.431, averaging 0.27 over the species. AMOVA partitioned over 80% of the total variation within populations. Multidimensional scaling revealed weak but significant differentiation into a northern and southern provenance. Despite selective sampling, the genetic data provided useful information for the management of Hemigenia exilis. For restoration, seed should be collected from a range of habitats of several populations, while keeping the two provenances separate. This strategy is likely to maintain high genetic diversity and locally adapted populations.     

Abundant genetic diversity in cultivated <Emphasis Type="Italic">Codonopsis pilosula</Emphasis> populations revealed by RAPD polymorphisms

Genetic diversity of seven cultivated populations of Codonopsis pilosula Nannf. from Longxi County, Gansu Province of China was estimated using randomly amplified polymorphic DNA (RAPD) markers. The 17 selected RAPD primers amplified 205 polymorphic bands out of a total of 235 (87.2%). Nei’s gene-diversity statistics and population differentiation parameters based on AMOVA analysis indicated that the cultivated C. pilosula populations remained a high level of genetic diversity with Hs = 0.299 and I = 0.450. A greater proportion of genetic diversity was found within (77%) rather than among (23%) the populations. In addition, we also detected that populations from different altitudes had a considerable genetic differentiation after 40 years of cultivation at the same site. Populations from higher altitude had lower genetic diversity than those from lower altitude. Our results suggested that irregular and sparse cultivation practices, i.e., random collecting, preserving, and planting seeds of the medicinal species without deliberate selection, might be an efficient way to conserve genetic resources of medicinal plants, in addition to their effective uses.     

Genetic diversity of anchote (Coccinia abyssinica (Lam.) Cogn.) from Ethiopia as revealed by ISSR markers

Anchote (Coccinia abyssinica) is plant endemic in Ethiopia with a high calcium content grown for its edible tuberous roots. In spite of its importance as food security crop, there is no information available on molecular genetic diversity of this crop. Therefore, the aim of this study was to assess genetic diversity within and among 12 populations of anchote using ISSR markers. Using nine ISSR primers, a total of 87 scorable bands was generated of which 74 were polymorphic. Within population diversity based on polymorphic bands ranged from 13.8 to 43.53 % with a mean of 33.05 %, Nei’s genetic diversity of 0.04–0.156 with a mean of 0.12, Shannon information index of 0.07–0.23 with a mean of 0.175 and analysis of molecular variation (AMOVA) of 51.4 % were detected. With all diversity parameters, the highest diversity was obtained from Gimbi, Bedele and Ale populations, whilst the lowest was from Manna. AMOVA showed a 48.56 % between populations variability and significantly lower than that of within population variation. Population differentiation with F_ST was 48.56 %. From Jaccard’s pairwise similarity coefficient, Decha and Nedjo were most related populations exhibiting 0.76 similarity and Manna and Nedjo were the most distantly related populations with similarity of 0.52. The only pentanucliotide primer used in the study, Primer 880 (GGAGA)3, showed a unique band in some individuals that appeared to be associated with morphological quantitative traits (lowest seed number, high protein content, largest fruit size and smallest vine length). Illubabor and Gimbi populations exhibited highest genetic diversity so that the populations should be considered as the primary sites in designing conservation areas for this crop.     

Ex situ genetic conservation of endangered Vatica guangxiensis (Dipterocarpaceae) in China

RAPD polymorphisms were applied to check the efficiency of ex situ genetic conservation of endangered Vatica guangxiensis X. L. Mo. endemic to southwestern China. Low level of genetic variation was revealed in three remaining natural populations. Twenty random primers, each with 10 base pairs, generated 231 bands with 53.68% being polymorphic, and with an average of 32.46% being polymorphic in each natural population. Strong population differentiation was revealed by AMOVA (analysis of molecular variance) and Gst value was 0.3764. The population ML ex situ conserved in the Xishuangbanna Tropical Botanical Garden contained an intermediate genetic variation compared with natural populations, with 30.74% bands being polymorphic. Of the total 231 bands generated in V. guangxiensis, 204 bands were also detected in population ML, indicating that 88.31% of the total genetic variations of this species were conserved in ex situ population. If only the alleles with moderate to high frequency (P>0.05) were considered, 204 out of 209 bands (97.61%) occurred in ex situ population ML. RAPD analysis also detected one exclusive band in natural population NS, and five in natural population NP, three of these exclusive bands were generated in every samples of natural population (NP), and other three had moderate to high frequencies. While none of these exclusive bands were detected in ex situ conserved population ML. Our conclusions are that the ex situ conserved population ML contains representative genetic variation to maintain long-term survival and evolutionary process of V. guangxiensis, and that more extensive ex situ sampling in natural population NS and NP is needed to conserve more exclusive alleles in ex situ population. The tropical area in the Botanical Garden would play a more important role in the ex situ conservation of rare and endangered plants.     

Genetic diversity of <Emphasis Type="Italic">Guizotia abyssinica</Emphasis> (L. f.) Cass. (Asteraceae) from Ethiopia as revealed by random amplified polymorphic DNA (RAPD)

Genetic diversity of 70 populations of niger (Guizotia abyssinica) representing all its growing regions in Ethiopia was investigated using random amplified polymorphic DNA (RAPD) to reveal the extent of its populations genetic diversity. Ninety-seven percent of the loci studied was revealed to be polymorphic for the whole data set. The within population diversity estimated by Shannon diversity index and Nei gene diversity estimates was revealed to be 0.395 and 0.158, respectively. The extent of genetic variation of populations from major niger producing regions was significantly lower than that of populations from other regions; however, it is distributed regardless of altitude of growth. Genetic differentiation between populations was estimated with Shannon index as G^′ _ST (0.432), Nei’s G _ST (0.242) and AMOVA based F _ST (0.350) and appears to be equivalent to the average values calculated from various RAPD based studies on outcrossing species. Higher proportion of the variation detected by AMOVA resided within populations (64.58%) relative to the amount of variation among populations (35.42%). UPGMA cluster analysis showed that most of the populations were clustered according to their region of origin. However, some populations were genetically distant from the majority and seem to have unique genetic properties. It is concluded that the crop has a wide genetic basis that may be used for the improvement of the species through conventional breeding and/or marker assisted selection. Collection of germplasm from areas not yet covered and/or underrepresented is the opportunity to broaden the genetic basis of genebank collection.     

Genetic variation in the threatened South American conifer Pilgerodendron uviferum (Cupressaceae), detected using RAPD markers

Pilgerodendron uviferum (Cipres de las Guiatecas, Cupressaceae) is a long-lived conifer, endemic to southern Chile and Argentina, reaching a southern limit in Tierra del Fuego. Remnant populations are generally fragmented and highly disturbed because of exploitation for timber, grazing and fire. The extent of genetic variation within and between 16 populations of this species, distributed throughout its range, was assessed using random amplified polymorphic DNA (RAPD) markers. Eight 10-mer primers produced a total of 84 scorable markers, 30 of which (35.7%) were polymorphic. AMOVA indicated that 18.6% of the variation recorded was attributable to differences between populations, a relatively high value compared with other conifers from the region. Pair-wise Phi_st comparisons between populations were all significant at P<0.05, with one exception, highlighting a high degree of population differentiation. Values of Shannon's diversity index (S) differed significantly among populations (P=0.002, ANOVA), values ranging from 0.337 to 0.716, suggesting that some populations are currently characterised by very low genetic variability. Current patterns of genetic variation were related to biogeographic history and human impact. The high degree of population differentiation recorded here highlights the need for additional conservation measures for this species, both in terms of incorporating further populations into the protected areas, and the restoration of severely degraded populations, to ensure their continued viability.     

湖南典型茶树地理种群遗传多样性研究

本文利用RAPD分子标记技术，对4个湖南典型茶树地理种群的240个单株的遗传多样性、种群内和种群间的遗传变异进行研究，结果表明：21个10碱基随机引物共检测到226条谱带，其中多态性谱带为201条，占88.9%。遗传多样性分析结果显示：Shannon's多样性指数为0.43，74.7%的变异分布于群体内，而种群间变异占了25.3%；Nei's指数群体总基因多样度(HT)为0.37，群体内平均基因多样度(HS)0.28，群体间的基因多样度(HST)0.09，群体Nei's基因分化系数（GST）为0.23，说明76.7%的变异存在于种群间，群体内的变异占了总变异的23.3%，与Shannon's多样性指数相比基本一致，均表明种群内有较丰富的遗传变异；种群间的基因流（Nm）为0.74，显示种群间的基因交流有限。     

Genetic structure among earthworms (Lumbricus terrestris L.) from different sampling sites in western Germany based on random amplified polymorphic DNA

Earthworms are being used as bio-indicators to assess terrestrial pollution. However, it is often not known whether their populations possess a uniform genetic structure, which would allow comparison of residues or biological properties of earthworms from different sampling locations. In order to investigate this point, random amplified polymorphic DNA (RAPD) variation was surveyed in earthworms (Lumbricus terrestris) from five different sampling sites in Germany. Forty oligonucleotide RAPD primers (10 base pairs in length) were screened, three of which produced high polymorphic band patterns. A total of 61 DNA fragments were detected in 90 individuals of L. terrestris from five sampling sites with 49 (80.3%) RAPD markers being polymorphic. The genetic similarities within (band sharing rates between 0.756 and 0.795) and among the L. terrestris populations (0.635) were similar even at widely separate locations. Inter-population variation in the RAPD pattern for all five earthworm populations accounted for 37.9% of the total variation, while intra-population variation for three adjacent Saarland populations accounted for only 18.0% of the total variation. Principal component analysis (PCA) and the genetic distances of the populations confirm these results. Twenty-four percent of the genetic distance is caused by geographical isolation as shown by a test for isolation by distance. These results show that L. terrestris fulfils the genetic qualifications for a bio-indicator particularly at closely located sampling sites. However, the results also suggest that earthworm studies of widely separated locations should include genetic characterisation of the earthworm samples.     

Genetic diversity in <Emphasis Type="Italic">Eremostachys superba</Emphasis> Royle ex Benth. (Lamiaceae), an endangered Himalayan species,as assessed by RAPD

Eremostachys superba Royle ex Benth. (Lamiaceae) has undergone a severe decline in population size since its discovery in the North-western Himalayas in late 19th century. One hundred and seventy-two plants from six populations in the Indian states of Uttar Pradesh and Jammu & Kashmir, located between 0.45 km and 455.72 km apart from each other were evaluated for RAPD polymorphism. Sixteen random primers generated 92 bands overall, 77 of which were polymorphic. Shannon’s index of genetic diversity within populations (H _o) ranged between 0.305 and 0.421; the average within-population diversity (H _pop) was 0.389; and the total species diversity (H _sp) was 0.478. The population from Mohand (representing the type locality) had the fewest plants, at 18, and was genetically the most depauperate. Among the other populations, ranging in size between 52 and 1,022 individuals, no relation between population size and genetic diversity was evident. It is suggested that these six populations represent relics of a larger, extended population, in which the presence of perennating rootstocks has helped preserve historic patterns of genetic diversity. AMOVA revealed that 83.01% of the variation exists within populations, which was consistent with earlier studies on the reproductive biology of E. superba, which indicated this species is predominantly allogamous. FST distances between all populations were significant, indicating geographic differentiation despite some of them being closely separated. Habitat restoration and protection from indiscriminate harvesting are proposed as primary strategies for conserving E. superba. Rejuvenation of the Mohand population through intrapopulation crossing between plants bearing diverse molecular phenotypes is also suggested.