硬核资源遗传多样性的AFLP分析

为了解云南省硬核[Scleropyrum wallichianum（Wight et Arn.）Arn.]的遗传多样性,采用AFLP标记分析了7个居群84份种质材料的遗传变异。结果表明,从64对引物组合中挑选出多态性较好的引物8对,共扩增出1 728条带,其中多态性条带1 388条,多态性百分率为80.14%。硬核在物种水平的多样性指数分别为Na=1.416,Ne=1.179,H=0.137,I=0.225,在居群水平上分别为H=0.111,I=0.175;在遗传相似性系数为0.52时,这些种质材料可分为3组,其中易武居群具有丰富的遗传变异,大部分的遗传变异存在于居群内,而在0.05置信区间内居群间遗传变异仅为11.5%;居群间的遗传距离和地理距离无显著相关性（r=0.0323,P=0.5820）。因此,硬核资源可采用就地和迁地保护策略,以增加其遗传多样性。     

黄芩种质资源ISSR遗传多样性的分析及评价

采用ISSR分子标记技术对6个野生或栽培居群共147份黄芩种质进行遗传多样性分析和评价。分析结果表明,51个ISSR引物中筛选出18条扩增条带清晰、重复性好和多态性高的引物,共扩增出485条清晰的条带,其中466条具有多态性,平均多态性位点比率为96.08%,平均Nei’s基因多样性指数和Shannon’s信息指数分别为0.244 4和0.388 9,等位基因数（Na）和有效等位基因数（Ne）分别为1.993 8和1.383 9,遗传分化指数G_st=0.122 3,遗传一致度（I）和遗传距离（D）分别为0.951 5和0.050 1,说明收集的黄芩种质资源在总体上具有较高的遗传多样性,不同居群间存在一定的遗传分化和基因交流,遗传变异主要存在于居群内。分子聚类结果表明,同一地区的种质并没有按照收集来源完全聚类,可能与种质不同起源或民间栽培引种有关。在DNA分子水平揭示黄芩种质资源的遗传多样性水平,将为进一步黄芩种质资源评价、保存和新品种选育等利用提供依据。     

野生扁蓿豆种质资源AFLP遗传多样性的分析

本研究采用AFLP标记对河北省、辽宁省、吉林省、山西省、内蒙古自治区和西藏自治区的10份野生扁蓿豆居群进行遗传多样性分析。从64对AFLP引物组合中,筛选出8对扩增条带清晰、多态性高的引物组合,8对引物共扩增出640个条带,其中多态性条带有472个,多态性位点百分率为73.8%;Nei's基因多样性指数平均为0.157,Shannon's多态性信息指数平均为0.099;居群间遗传相似系数(GS)平均值为0.827。聚类和主成分分析可将10个扁蓿豆居群聚为3大类,结果与居群的地理分布大致相符,呈一定的地域性分布规律。由此可见,AFLP分子标记研究结果能较好地揭示扁蓿豆居群间的遗传多样性。对我国各地的野生扁蓿豆资源的广泛收集、评价和基因资源的保护有重要的意义。     

广西地不容种质资源的ISSR分析

采用简单序列重复区间扩增(ISSR)分子标记技术对广西地不容3个野生居群和1个引种居群共92个个体进行了遗传多样性研究。10个引物共扩增出61条带,其中60条具多态性,多态性位点百分率为98.36%。4个居群多态性百分率在73.77%～86.89%。Nei’s基因多样性指数(H)为0.3379,Shannon信息多样性指数(Ⅰ)为0.5055。3个野生居群Nei’s遗传分化系数(Gst)表明:83.87%遗传变异分布在居群内,16.13%的遗传变异分布在居群间。引种居群与3个野生居群间的遗传一致度达0.8846。引种居群有效地保护了广西地不容的遗传多样性。     

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。     

硬核资源遗传多样性的AFLP分析（英文）

为了解云南省硬核[Scleropyrum wallichianum (Wight et Arn.) Arn.]的遗传多样性,采用AFLP标记分析了7个居群84份种质材料的遗传变异。结果表明,从64对引物组合中挑选出多态性较好的引物8对,共扩增出1 728条带,其中多态性条带1 388条,多态性百分率为80.14%。硬核在物种水平的多样性指数分别为Na=1.416, Ne=1.179, H=0.137, I=0.225,在居群水平上分别为H=0.111,I=0.175;在遗传相似性系数为0.52时,这些种质材料可分为3组,其中易武居群具有丰富的遗传变异,大部分的遗传变异存在于居群内,而在0.05置信区间内居群间遗传变异仅为11.5%;居群间的遗传距离和地理距离无显著相关性(r=0.0323, P=0.5820)。因此,硬核资源可采用就地和迁地保护策略,以增加其遗传多样性。     

迁地保护的金花茶遗传多样性评价

采用ISSR分子标记技术对迁地保护的金花茶3个人工栽培居群进行遗传多样性分析。揭示了迁地保护金花茶的遗传多样性水平和结构。11条引物共扩增出87条带,其中36条具有多态性,多态位点比例(P)为41.38%。单个居群的多态带百分率P从21.84%到25.29%,平均值为24.14%。在物种水平上,期望杂合度和Shannon信息指数分别为0.1525和0.2273;在居群水平上:期望杂合度和Shannon信息指数分别为0.1069和0.1528。通过与野生居群遗传参数比较,表明该种质圃基本有效地保护金花茶的遗传多样性总水平。遗传分化系数为0.2990。表明迁地保护金花茶遗传变异发生在居群内的个体间占70.10%、29.90%的遗传变异发生在居群间。     

基于SSR和ISSR的鄱阳湖流域野生菰资源的遗传多样性分析

用SSR和ISSR标记对鄱阳湖流域30个野生菰居群的遗传多样性与遗传结构进行了分析。筛选出的19对SSR引物共扩增出多态性条带253条,平均多态性条带比率(PPB)为91.67%,Nei's基因多样性指数(He)和平均Shannon信息指数(I)分别为0.2712和0.4144,遗传相似性系数(GS)为0.5590~0.8368,遗传距离(GD)的变化范围为0.1632~0.4410;筛选出的14个ISSR标记引物共扩增出83条条带,平均多态性条带比率(PPB)为78.29%,Nei's基因多样性指数(He)和平均Shannon信息指数(I)分别为0.2386和0.4174,遗传相似性系数(GS)为0.5132~0.9342,遗传距离(GD)变化范围为0.0658~0.4868。根据SSR和ISSR基因型数据,采用UPGMA法分别在阈值为0.698和0.728时可将30个野生菰居群聚为3类。可能受人为、水流、动物活动、风等多种因素的影响,居群间的亲缘关系与地理分布无明显相关性。本研究表明,鄱阳湖流域野生菰居群间SSR和ISSR基因型的多样性丰富,居群间的这种遗传差异或变异,对该地区乃至更大范围内野生菰的遗传进化、基因资源的开发利用和种质资源的保护有着重要意义。     

野生毛樱桃SSR遗传多样性和遗传结构分析

利用11对SSR多态性引物对秦岭山脉5个居群11个亚居群104份野生毛樱桃材料进行遗传多样性和遗传结构分析,以明确中国樱桃的种质资源,为樱桃育种以及生产中改良樱桃砧木提供理论依据。结果显示:(1)11对引物共检测出110个等位位点,各引物扩增的多态性条带在5~13个之间;观察到的平均等位基因数(A)为10,有效等位基因数(Ae)为7.586 3。(2)秦岭山脉天水居群的多态性比率(PPB)为69.55%、长安居群为76.53%、太白居群为81.36%、华阴居群为85.00%、眉县居群为92.17%,5个居群总的多态性比率为81.62%,各位点平均期望杂合度(He)为0.864 0。(3)秦岭山脉野生毛樱桃各居群的基因分化系数(Gst)为0.081 2,居群内的遗传分化占91.88%,居群间的遗传分化占8.12%;基于Gst测得基因流(Nm)为2.827 3,秦岭山脉Nei遗传距离为0.489 7,香农信息指数为2.101 9。研究认为,秦岭山脉野生毛樱桃的遗传多样性水平较高,遗传分化主要存在于居群内,且基因交流没有受到阻碍,居群内现存的遗传多样性水平对遗传漂变不敏感,但居群间存在适中的基因交流可能会降低整个种群的再分化。     

基于RAPD标记的南苍术居群遗传多样性分析

采用RAPD标记技术对分布于江苏小九华山、小汤山和湖山,安徽金寨和芜湖以及湖北保康和英山的7个南苍术〔Atractylodes lancea(Thunb.)DC.〕野生居群的28个单株基因组总DNA进行PCR扩增,在此基础上分析居群的遗传多样性及遗传分化,并采用聚类分析法对居群的遗传关系进行分析。结果表明:用18条RAPD引物共扩增出193条带,其中多态性条带111条,多态性条带百分率(PPB)为57.51%;平均每条引物扩增出10.72条带,其中多态性条带6.17条。从省级水平看,安徽居群的PPB、有效等位基因数(Ne)、Nei’s基因多样性指数(H)和Shannon信息指数(I)均最低,而湖北居群的Ne、H和I均最高,但江苏居群的PPB最高;从居群水平看,湖北保康居群的PPB、Ne、H和I均最高,而安徽金寨居群均最低。7个居群的基因分化系数和基因流分别为0.206 5和1.921 5,说明7个居群总遗传变异的20.65%存在于居群间、79.35%存在于居群内。7个居群间的遗传距离为0.150 7～0.252 1,其中,安徽金寨和芜湖居群间最小(0.150 7),江苏湖山和安徽芜湖居群间最大(0.252 1)。基于遗传距离的聚类分析结果表明:7个居群可分为2组,湖北保康居群单独成组,其他6个居群聚为另一组;来自同一居群的单株均聚在一起。研究结果提示:南苍术居群间的遗传多样性较低,居群间无明显的遗传分化。     

广东省猴耳环遗传多样性研究

为了解猴耳环(Archidendron clypearia)种质资源的遗传多样性,以广东省12个野生猴耳环群体的146份种质资源为材料,采用SSR分子标记技术对其遗传多样性和亲缘关系进行分析。结果表明,21对SSR引物共检测到249个等位基因,平均每对SSR引物检测的等位基因数(Na)为11.857,有效等位基因数(Ne)为3.500,期望杂合度(He)为0.718,多态信息含量(PIC)为0.676;12个群体中博罗群体的Shannon多样性指数(I=0.528)和有效等位基因数(Ne=0.716)均最大,是遗传多样性最丰富的群体;群体间的遗传分化系数为0.071,AMOVA分析表明,猴耳环的遗传变异主要在群体内(97%),群体内的遗传分化大于群体间。聚类分析表明,遗传系数在0.16时,可将12个群体分为6大类,与主坐标分析的结果大致相同。这为发掘、利用与保护猴耳环群体种质资源,开展猴耳环优良品种的遗传育种提供重要的理论依据。     

Genetic relationships among South-East Turkey wild barley populations and sampling strategies of Hordeum spontaneum

To assess the genetic diversity and the genetic structure of Turkish wild barley (Hordeum spontaneum Tell.) populations, 76 genotypes from ten ecologically and geographically different locations were analyzed by means of amplified fragment length polymorphism (AFLP) markers. Five primer combinations produced 187 scorable bands, of which 117 (62.6%) were polymorphic. Several population-specific and genotype-specific bands were identified, which differentiate populations or genotypes. Genetic distance, determined by Nei’s distance coefficient, varied from 0.07 to 0.21 with an average of 0.13. In the UPGMA dendrogram based on Nei genetic distances, the Hordeum spontaneum populations were separated into two major clusters. Genetic diversity was larger among (68%) than within (32%) populations. Eight AFLP bands were strongly correlated to the altitude of the collecting site, while no clear trend was detected between geographical origin and genetic diversity. Our results strongly suggest the need for a change in Hordeum spontaneum sampling strategy: more populations, rather then more individuals within population, should be sampled to appraise and safeguard genetic diversity in the wild barley gene pool.     

AFLP Analysis of Genetic Diversity of the Endangered Species Sinopodophyllum hexandrum in the Tibetan Region of Sichuan Province,China

Amplified fragment length polymorphism (AFLP) markers were used to estimate the genetic diversity of seven wild populations of Sinopodophyllum hexandrum (Royle) Ying from the Tibetan region of Sichuan Province, China. Six primer combinations generated a total of 428 discernible DNA fragments, of which 111 were polymorphic. The percentage of polymorphic bands (PPB) was 25.93 at the species level, and PPB within population ranged from 4.91 to 12.38%. Genetic diversity (H _E) within populations varied from 0.01 to 0.04, averaging 0.05 at the species level. As revealed by the results of AMOVA analysis, 58.8% of the genetic differentiation occurred between populations, and 41.2% within populations. The genetic differentiation was, perhaps, due to the limited gene flow (N _m=0.43) of the species. The correlation coefficient (r) between genetic and geographical distance using Mantel's test for all populations was 0.698 (P=0.014). The UPGMA cluster analysis revealed a similar result in that the genetic distances among the populations show, to a certain extent, a spatial pattern corresponding to their geographic locations. On the basis of the genetic and ecological information, we propose some appropriate strategies for conserving the endangered S. hexandrum in this region.     

西南地区6种魔芋属植物基于cpDNA序列的遗传多样性研究

研究野生作物资源的遗传变异及分化机制对种质资源的收集与改良具有重要意义。魔芋是我国西南地区的特色经济作物,但由于受到人为活动干扰,野生种群不断衰退。为评估西南地区魔芋属(Amorphophallus)野生群体的遗传多样性,探究代表性物种的系统发育地位,该研究利用3个叶绿体DNA(cpDNA)片段,分析了魔芋6个物种的遗传多样性,重建了种间系统发育关系。结果表明:(1)西南地区野生魔芋群体的遗传多样性普遍较低,虽然单倍型多样性(H_d)均值为0.428,但近一半群体只有1个单倍型,6个物种整体水平上的单倍型多样性在0.704到0.983之间。(2)在6个物种间检测到高水平的遗传分化,遗传分化系数(F_ST)值在0.481到0.967之间。(3)系统发育分析表明,选取的27个魔芋种主要聚成3个分支:非洲分支、东南亚分支和东亚大陆分支。疣柄魔芋(A. paeoniifolius)隶属于东南亚分支,而东亚大陆分支A包含花魔芋(A. konjac)和西盟魔芋(A. krausei),东亚大陆分支B由东亚魔芋(A. kiusianus)、滇魔芋(A. yunnanensis)和东京魔芋(A. tonkinensis)构成。生境隔离与人为干扰造成了西南地区野生魔芋群体较低的遗传多样性,魔芋属东亚大陆分支的分化可能与早期的快速扩张和生态适应有关。该研究为西南地区魔芋资源的合理保护、可持续利用和杂交育种提供了参考资料。     

Genetic Variability and Genetic Structure of Wild and Semi-domestic Populations of Tasar Silkworm (Antheraea mylitta ) Ecorace Daba as Revealed through ISSR Markers

The genetic diversity in the wild and semi-domestic populations of Daba ecorace of Antheraea mylitta was studied to ascertain the distribution of variability within and among populations of semi-domestic bivoltine (DB), trivoltine (DT) and nature grown wild populations (DN) with inter-simple sequence repeat (ISSR) markers. A total of 138 markers were produced among 56 individuals of the three populations, of which 98% were polymorphic. For the individual populations, the percentage polymorphism was 58.69, 52.9 and 77.54 for DB, DT and DN, respectively. Average number of observed (1.791 ± 0.408) and effective alleles (1.389 ± 0.348) was also high in the wild populations in comparison to the bivoltine and trivoltine semi-domestic populations. Genetic diversity (H_t) in DB, DT and DN was 0.180 ± 0.033, 0.153 ± 0.032 and 0.235 ± 0.033, respectively and within-population genetic diversity (H_s) ranged from 0.166 to 0.259 with a mean of 0.189. Mean gene differentiation (G_ST) was found to be 0.25. Shanon’s diversity index was 0.278, 0.237 and 0.361 for DB, DT and DN and overall it was 0.391. Gene flow (N_m) among the populations was 1.509. The dendrogram produced by UPGMA with Dice’s genetic distance matrices resulted in the formation of three major clusters separating the three populations. Considerable intra- and inter-population variability is found in all three populations. The population structure analysis further suggests that the semi-domestic populations of Daba ecorace are at the threshold of differentiating themselves. The high genetic variability present within wild Daba population of A. mylitta is of much importance for conservation as well as utilization in systematic breeding program.     

双孢蘑菇遗传多样性分析

应用AFLP指纹技术对双孢蘑菇的20个野生菌株和5个栽培品种的遗传多样性进行了研究。AFLP指纹揭示出20个野生异核体菌株所固的基因型。5个商业品种表现出比较一致的AFLP指纹,但也显示出它们之间的一些差别。由单孢分离获得的同核体菌株携带着部分异核体菌株的AFLP指纹;由同一子实体分离得到的大部分单孢菌株是异核体菌株,它们具有与其亲本一致的AFLP指纹。UPGMA分析揭示出2个与地理分布（美洲、欧洲）和相对应的组。研究结果表明：（1）在野生菌株之间存在着明显的遗传差异;（2）大多数单孢分离的菌株具有与母本一致的遗传物质;（3）野生菌株间的遗传变异大于栽培品种间的变异;（4）在双孢蘑菇的遗传多样性分析中,AFLP技术是非常有效的工具。     

珊瑚菜居群遗传多样性的SRAP分析

利用SRAP对伞形科单种属珊瑚菜7个野生居群和1个栽培居群进行了研究。结果表明:共筛选出8个引物组合,在珊瑚菜8个居群中共扩增出168条条带,其中多态性条带为118条,多态性比率为70.23%;平均每对引物扩增的多态性条带为14.75。各居群之间珊瑚菜遗传相似性系数范围为0.8306~0.9836,遗传距离范围为0.0165~0.1856。聚类分析表明,以相似性系数大于0.8并结合地理分布来看,所研究的野生珊瑚菜居群可以大体分为3类,辽宁大连的野生居群为一类,山东威海—山东青岛的居群为一类,而山东日照—广州深圳之间的为一类。     

Genetic diversity and phylogenetic relationships in alder,Alnus firma, revealed by AFLP

Molecular markers for alder,Alnus firma Sieb. et Zucc, have not been studied extensively. Here, we used amplified fragment length polymorphism (AFLP) to investigate genetic relationships among 15 natural populations. EcoRI-ACG + Msel-CTG combinations revealed the highest polymorphism (62.2%). A total of 171 DNA fragments were identified. On average, 58.1% of the AFLP markers that were generated using four primer pairs were polymorphic. Diversity was insignificant among the populations. The combination of a wind-pollinated, outcrossing breeding system along with large population sizes, and the ability to regenerate by stump sprouting may explain the high level of genetic diversity within this species. The majority (98%) of the genetic variance resided within populations. The average number of individuals that were exchanged between populations per generation was very high (N _em = 12.3). Gene dispersal in alder is apparently by seed dispersalvia water and human activity as well as through pollen. Five individuals per population were claded in the same cluster.     

用扩增片段的长度多态性分子标记探讨盾叶薯蓣的遗传多样性

用扩增片段的长度多态性(amplified fragment length polymorphism,AFLP)标记分析研究了中国5个盾叶薯蓣居群30个个体的遗传多样性。筛选出9对AFLP引物,从中检测到14698条清晰可见的条带,其中多态性带12628条,多态性比率85.92%。Shannon信息指数(I)为0.3656±0.1721,物种水平的Nei基因多样性(H)为0.2322±0.2200。遗传变异分析表明,物种水平的遗传分化系数Gst为0.4827,说明其群体间存在一定的遗传分化,居群间的基因流Nm为0.5358,居群间遗传交换较小。聚类分析结果显示5个居群盾叶薯蓣有较为丰富的遗传变异,且与地理分布有相关性。     

资源昆虫角倍蚜遗传多样性及遗传分化的AFLP分析

本研究采用AFLP分子标记技术对资源昆虫角倍蚜Schlechtendalia chinensis 6个种群共102个个体的样本进行遗传多样性分析, 探讨角倍蚜主要分布区不同种群之间的遗传分化及其变异程度, 为合理利用和保护该经济昆虫提供分子方面的证据。结果表明: 经过摸索实验筛选出的4对选择性扩增引物共扩增条带126条, 多态性条带比例为100%, 种群多态性位点比例介于23.81%～66.67%之间; Nei's基因多样性指数介于0.0942～0.1980之间; Shannon's多样性数介于0.1381～0.3027之间; AMOVA分析显示57.99%的变异来源于种群内, 42.41%的变异来源于种群间（P<0.01）; 总群体的Fst值为0.4242; NJ聚类树显示角倍蚜6个种群共形成两个大的聚类簇, 阳雀、丹寨和汉中种群聚为一支, 安县、竹山和龙胜3个种群聚为另一支。总体上, 角倍蚜种内的遗传多样性较低, 而种群间的遗传分化较大, 遗传距离与地理距离之间没有显著的相关关系（P>0.05）。