岷江流域王百合和通江百合多酚氧化酶同功酶的研究

应用聚丙烯酰胺凝胶垂直平板电泳技术,对岷江流域王百合3个采样点的14个样品和通江百合4个采样点的4个样品的多酚氧化酶(PPO)同功酶进行了研究。结果表明,两种百合的多酚氧化酶在种间和种内均有一定差异。酶谱的聚类分析在遗传相似系数约0.5处,将两种百合18个样品分成5个类群,其中王百合的13个样品聚成了4类,样品12则与通江百合的4个样品聚成了1类;5个类群在外部形态上也呈现出相似性。     

ISSR标记技术及其在遗传多样性研究中的应用

ISSR(Inter-Simple Sequence Repeat)技术是在PCR中直接使用微卫星序列进行DNA扩增的一种DNA分子标记。章主要介绍了ISSR标记的原理、方法、特点及其在遗传多样性研究中的应用。ISSR标记方法具有无需知道任何靶标序列的微卫星背景信息、遗传多态性高、检测快速等特点,在遗传多样性研究中具有广泛的应用前景。     

Assessment of Genetic Diversity in Ziziphus mauritiana Using Inter-Simple Sequence Repeat Markers

Genetic diversity among 47 ber accessions belonging to cultivated species (Ziziphus mauritiana Lam) and one wild accession of Ziziphus nummularia (Burm F) Willed was investigated using Inter-Simple Sequence Repeat (ISSR) markers. A total of 167 amplification products were detected with 18 ISSR primers of which 152 (89.96%) were polymorphic. Most of the primers that produced distinct bands (14 primers out of 18) contained dinucleotide repeats. Primers based on (AC)n and (AG)_n repeats produced more polymorphic bands. Genetic similarity ranging from 43.07% to 90.30% suggested that the 48 Ziziphus genotypes used in the study were divergent. Cluster analysis based on UPGMA method and Bootstrap analysis separated all the 48 genotypes in four distinct clusters. The present study has successfully distinguished morphologically similar genotypes that emphasize the use of molecular markers to the taxonomists. Morphologically similar but genetically distinct genotypes, identified using ISSR markers could be potential sources for genotype identification and to resolve controversies over misnomination of ber genotypes. Present study is the first report on the exploitation of ISSR markers in ber for genetic diversity analysis.     

白沙蒿不同地理种群遗传分化的ISSR分析

运用ISSR技术,对白沙蒿的5个种群进行遗传分化的分析。12个引物在108个个体中共扩增出222个位点,其中,多态位点为218个,多态位点百分率为98.20%,Shannon多样性指数为0.315 4,具有高的多态性。种群间的遗传分化系数（G_st）为0.076 7,与AMOVA分析的结果一致（Ф_st为7.96%）,表明绝大多数的遗传变异存在于种群内部。各种群间遗传一致度高达98%以上,遗传距离很小,基因流达3.008 2,均表现出白沙蒿各种群存在着广泛的基因交流,有着很小的遗传分化。     

Genetic Diversity of Kenaf (Hibiscus cannabinus) Evaluated by Inter-Simple Sequence Repeat (ISSR)

Understanding genetic diversity is very useful for scientific utilization for breeding. In this study, we estimated the genetic distances in a panel of 84 kenaf accessions collected from 26 countries and regions using ISSR markers. The results of UPGMA analysis showed that kenaf germplasm had abundant genetic variation, with genetic dissimilarity coefficients ranging from 0.01 to 0.62. The in-group dissimilarity coefficient (0.29) was observed in 84 kenaf accessions, and all the accessions could be divided into three groups: cultivars (L_1–1), relatively wild species (L_1–2 and L_1–3), and wild species (the others). Further in-group analysis in group L_1–1 (0.19) revealed that the kenaf cultivars could be divided into five subgroups with distinct regional characteristics. It is imperative that genes be exchanged among all kinds of tested varieties from different origins. The results provide a useful basis for kenaf germplasm research and breeding.     

特有珍稀濒危植物景东报春遗传多样性的ISSR分析(英文)

景东报春(Primula interjacens Chen)是一分布在中国云南中南部的稀有的、分布区狭窄的特有种。该种含2个变种:原变种景东报春(P.interjacens Chen var.interjacens),现存一个居群;和光叶景东报春(P.interjacens Chen var.epilosa C.M.Hu),有2个居群。本文用ISSR分子标记对景东报春现存的3个居群进行了遗传多样性分析。和我们最初的预计相反,尽管居群小、分布区狭窄,景东报春的遗传多样性却很高(在居群水平的多态位点率P=59.75％,基因多样性H_E=0.2368,Hpop=0.3459:在种的水平的多态位点率P=75.47％,遗传多样性H_T=0.3205,总的遗传多样性Hsp=0.4618)。推测异型花和杂交可能是维持高的遗传多样性的原因。Nei的遗传多样性水平(26.13％)和Shannon多样性指数(25.09％)显示居群间存在中度分化。尽管景东报春原变种和变种在形态上有区别,但用非加权配对算数平均法(UPGMA)聚类分析的结果表明,两者并没有明显的遗传分化,可处理为一个单一的分类单元。基于观察到的居群遗传信息,建议采取就地保护和迁地保护的保护措施。     

特有珍稀濒危植物景东报春遗传多样性的ISSR分析

景东报春(Primula interjacens Chen)是一分布在中国云南中南部的稀有的、分布区狭窄的特有种.该种含2个变种:原变种景东报春(P.interjacens Chen var.interjacens),现存一个居群;和光叶景东报春(P.interjacens Chen var.epilosa C.M.Hu),有2个居群.本文用ISSR分子标记对景东报春现存的3个居群进行了遗传多样性分析.和我们最初的预计相反,尽管居群小、分布区狭窄,景东报春的遗传多样性却很高(在居群水平的多态位点率P=59.75%,基因多样性HE=0.236 8,Hpop=0.345 9;在种的水平的多态位点率P=75.47%,遗传多样性HT=0.320 5,总的遗传多样性Hsp=0.4618).推测异型花和杂交可能是维持高的遗传多样性的原因.Nei的遗传多样性水平(26.13%)和Shannon多样性指数(25.09%)显示居群间存在中度分化.尽管景东报春原变种和变种在形态上有区别,但用非加权配对算数平均法(UPGMA)聚类分析的结果表明,两者并没有明显的遗传分化,可处理为一个单一的分类单元.基于观察到的居群遗传信息,建议采取就地保护和迁地保护的保护措施.     

利用SSR分子标记分析秦岭冷杉自然居群的遗传多样性

利用10对SSR引物对濒危植物秦岭冷杉(Abies chensiensis)6个自然居群的120个个体进行了遗传多样性研究,旨在分析秦岭冷杉6个自然居群的遗传多样性、遗传结构及基因流变化.研究结果表明,120个个体在10个位点上共检测到149个等位基因,平均每个位点的等位基因数(A)为14.9,每个位点的有效等位基因数(e)为7.7,每个位点的平均预期杂合度(He)和平均观察杂合度(Ho)分别为0.841和0.243,Shannon多样性指数(Ⅰ)为2.13,自然居群杂合性基因多样度的比率(FsT)为6.7％,居群间的基因流(Nm)为3.45.利用Mantel检测到自然居群的遗传距离与地理距离间无显著相关性(r=0.490 6,P＞0.05).秦岭冷杉自然居群的遗传多样性水平较低,遗传变异主要存在于居群内部.结合研究数据、实地调查及相关资料,推测秦岭冷杉自然居群间基因流较原来增大可能是因为居群间发生了远交衰退.     

栗属中国特有种居群的遗传多样性及地域差异

Genetic variability in 30 populations of three endemic Castanea (Tourn.) L. species (C. moUissima BI., C. seguinii Dode. and C. henryi (Skan) Rehd. et Wils. ) in China was investigated using isoelectric focusing in thin-layer polyacrylamide slab gels at 20 loci coding for 12 enzyme systems. C. mollissima was found to possess a significant higher value of genetic variability than that of the other two species. The percentage of polymorphic loci (P) and the expected heterozygosity (He) were 90.0% and 0. 311 at the species level, respectively; while P = 84.7 %, He = 0.295 at the population level. Comparison of the population genetic variability of C. mollissima in three regions of China, the populations from the Yangtze River valley showed a markedly higher mean expected gene heterozygosity. The results provided additional evidence that the Yangtze River valley, particularly in Shennongjia area, was the center of the genetic diversity of C. moUissima. Genetic relationships among populations and species were assessed by Nei genetic identity (I) and standard genetic distance (D), suggesting that C. mollissima and C. seguinii have a closer relationship, and genetic distances were correlated with geographical distances among populations. Genetic differentiation between populations in C. mollissima, C. seguinii and C. henryi was 7.5%, 10.9% and 22.1%, respectively, and the gene flow rate (Nm) was 3.20, 2.05 and 0.88 respectively. The study provides useful imfonnation for understanding of the origin and evolutionary events in genus Castanea and planning an effective conservation strategy.     

Genetic Diversity and Phylogenetic Relationships in the Genus Lycopersicon (Tourn.) Mill. as Revealed by Inter-Simple Sequence Repeat (ISSR) Analysis

Inter-simple sequence repeat (ISSR) analysis was for the first time used to study the genetic diversity and phylogenetic relationships in 54 wild accessions and cultivars of the genus Lycopersicon. Analysis involved 14 ISSR primers homologous to microsatellite repeats and containing additional selective anchor nucleotides. In total, 318 ISSR fragments were amplified for the wild and cultivated tomato genomes. The interspecific polymorphism revealed with the ISSR primers was 95.6%. Species-specific ISSR fragments were detected for each tomato species. The highest number (more than 20) of species-specific fragments were obtained for L. esculentum sensu lato, although the intraspecific variation of ISSR patterns was low. UPGMA cluster analysis was used to construct a dendrogram and to estimate the genetic distances between the species of the genus Lycopersicon; between populations ofL. peruvianum, L. pimpinellifolium, and L. esculentum; and between tomato cultivars. The ISSR-based phylogeny was generally consistent with Lycopersicon taxonomy based on morphological and molecular evidence, suggesting the applicability of ISSR analysis for genotyping and phylogenetic studies in tomato.     

Multilocus Sequence Analysis of Nectar Pseudomonads Reveals High Genetic Diversity and Contrasting Recombination Patterns

The genetic and evolutionary relationships among floral nectar-dwelling Pseudomonas ‘sensu stricto’ isolates associated to South African and Mediterranean plants were investigated by multilocus sequence analysis (MLSA) of four core housekeeping genes (rrs, gyrB, rpoB and rpoD). A total of 35 different sequence types were found for the 38 nectar bacterial isolates characterised. Phylogenetic analyses resulted in the identification of three main clades [nectar groups (NGs) 1, 2 and 3] of nectar pseudomonads, which were closely related to five intrageneric groups: Pseudomonas oryzihabitans (NG 1); P. fluorescens, P. lutea and P. syringae (NG 2); and P. rhizosphaerae (NG 3). Linkage disequilibrium analysis pointed to a mostly clonal population structure, even when the analysis was restricted to isolates from the same floristic region or belonging to the same NG. Nevertheless, signatures of recombination were observed for NG 3, which exclusively included isolates retrieved from the floral nectar of insect-pollinated Mediterranean plants. In contrast, the other two NGs comprised both South African and Mediterranean isolates. Analyses relating diversification to floristic region and pollinator type revealed that there has been more unique evolution of the nectar pseudomonads within the Mediterranean region than would be expected by chance. This is the first work analysing the sequence of multiple loci to reveal geno- and ecotypes of nectar bacteria.     

贵州野生猕猴桃表型遗传多样性分析

通过对146份贵州猕猴桃种质资源表型遗传多样性的研究,为贵州猕猴桃品种选育打下基础,对146份不同区域猕猴桃种质资源48个质量性状进行描述、赋值,并结合单果重等9个数量性状,使用主成分分析、聚类分析等进行综合评价。结果表明,57个质量性状和数量性状均存在丰富的多样性和变异性;146份资源的Shannon’s遗传多样性指数为1.28,从各区域遗传多样性指数可以看出,演化进程上从遗传多样性指数高的地区向低的地区进行演化;主成分分析结果显示,前14个主成分累计贡献率达75.076%,其中前6个主成分的累计贡献率达51.911%,为提高品种选育和评价的工作效率,可将猕猴桃种质资源的描述指标简化为新梢被毛、果实被毛等18个性状;聚类结果显示,贵州猕猴桃种质的欧氏距离为1.00~25.00之间,在D=19.25处,146份野生猕猴桃分为6个类群。本文有效揭示了贵州省猕猴桃种质资源丰富的多样性和亲缘关系,明确野生猕猴桃各种群间性状区分特征和部分优良资源性状的地理分布,对贵州猕猴桃种质资源的种质鉴定、保护利用和品种选育有重要意义。     

中国东南沿海15个秋茄种群遗传多样性的SRAP分析

为进一步保护和恢复红树林资源提供分子方面的基础资料和科学依据,本研究采用SRAP标记对15个中国东南沿海红树植物秋茄种群的亲缘关系进行了分析.从120对参试引物组合中选出46对重复性好、条带清晰的引物组合对供试的15份材料基因组DNA进行PCR扩增,得到大小在50～1 000 bp之间条带270,其中多态性条带107条,多态性位点率为39.63%.15个秋茄种群的遗传相似系数在0.004～0.845之间,平均为0.412,说明中国东南沿海秋茄种群存在较丰富的遗传多样性.聚类分析把15份供试种群材料划分为4个类群.15个秋茄种群的遗传多样性没有明显的地域性差异,同一样地域的不同种群之间具有遗传上的差别.建议在今后我国的红树林保护中,应加强对海南东寨港、深圳福田、湛江东北大堤和湛江附城的秋茄种群保护.     

秦巴山区野板栗居群遗传多样性AFLP分析

利用荧光AFLP标记技术对来自秦巴山区的野板栗10个居群共262个单株进行遗传多样性研究.10对AFLP引物共扩增出1297条谱带,其中多态性位点数1011个,多态位点百分率为77.95%;Nei's基因多样性指数为0.1439～0.2046,总体为0.2518;Shannon信息指数的变异范围为0.1972～0.2895,总体为0.4089;甘肃地区野板栗居群遗传多样性水平最高,陕西宝鸡居群的遗传多样性水平最低.AMOVA分析表明野板栗居群闻的遗传变异占总变异的17.51%,居群内变异占69.76%.UPGMA聚类可将供试10个居群划分为3类,聚类结果表现出明显的地域性.     

High Genetic Diversity of Newcastle Disease Virus in Wild and Domestic Birds in Northeastern China from 2013 to 2015 Reveals Potential Epidemic Trends

Newcastle disease (ND), caused by the virulent Newcastle disease virus (NDV), is one of the most important viral diseases of birds globally, but little is currently known regarding enzootic trends of NDV in northeastern China, especially for class I viruses. Thus, we performed a surveillance study for NDV in northeastern China from 2013 to 2015. A total 755 samples from wild and domestic birds in wetlands and live bird markets (LBMs) were collected, and 10 isolates of NDV were identified. Genetic and phylogenetic analyses showed that five isolates from LBMs belong to class I subgenotype 1b, two (one from wild birds and one from LBMs) belong to the vaccine-like class II genotype II, and three (all from wild birds) belong to class II subgenotype Ib. Interestingly, the five class I isolates had epidemiological connections with viruses from southern, eastern, and southeastern China. Our findings, together with recent prevalence trends of class I and virulent class II NDV in China, suggest possible virus transmission between wild and domestic birds and the potential for an NDV epidemic in the future.     

云南栽培稻种SSR遗传多样性比较

采用64个SSR标记对96份云南水稻（Oryza sativa）地方品种和选育品种的遗传多样性进行比较分析。结果发现64个标记都具有多态性,共检测到741个等位基因,每个多态性位点检测到的等位基因数为2—29个,平均11．57个：Nei基因多样性指数（He）范围在0．345（RM321）-0．932（RM1）之间,平均为0．56。水稻品种的遗传多样性并非按地理位置均匀分布,而是在相似系数为0．17的水平上明显分为2个不同类群,即籼稻类群和粳稻类群,且籼粳亚种间的SSR多样性差异不明显,籼稻平均等位基因数（Ap）和Nei基因多样性指数（Ap=10．6,He=0．46）与粳稻品种（Ap=10．7,He=0．48）十分接近,可能与这些品种间存在一定频率的基因交流有关。糯稻和非糯稻在籼稻群和粳稻群中都有表现,没有特别的分布规律。云南栽培稻选育品种与地方稻亲缘关系较近,其遗传基础可能来源于云南水稻地方品种。本研究结果表明,SSR标记能较好地区分云南栽培稻品种,且云南水稻地方品种遗传多样性丰富,存在大量的优质性状可供育种实践选择。     

云南栽培稻种SSR 遗传多样性比较

采用64个SSR标记对96份云南水稻(Oryz a sativa)地方品种和选育品种的遗传多样性进行比较分析。结果发现64个标记都具有多态性, 共检测到741个等位基因, 每个多态性位点检测到的等位基因数为2-29个, 平均11.57个; Nei基因多样性指数(He)范围在0.345(RM321)-0.932(RM1)之间, 平均为0.56。水稻品种的遗传多样性并非按地理位置均匀分布, 而是在相 似系数为0.17的水平上明显分为2个不同类群, 即籼稻类群和粳稻类群, 且籼粳亚种间的SSR多样性差异不明显, 籼稻平均等位基因数(Ap)和Nei基因多样性指数(Ap=10.6, He=0.46)与粳稻品种(Ap=10.7, He=0.48)十分接近, 可能与这些品种间存在一定频率的基因交流有关。糯稻和非糯稻在籼稻群和粳稻群中都有表现, 没有特别的分布规律。云南栽培稻选育品种与地方稻亲缘关系较近, 其遗传基础可能来源于云南水稻地方品种。本研究结果表明, SSR标记能较好地区分云南栽培稻品种, 且云南水稻地方品种遗传多样性丰富, 存在大量的优质性状可供育种实践选择。     

西南地区硬枝野荞麦(Fagopyrum urophyllum)居群的遗传多样性研究

采用等位酶电泳技术了云南省中北部昆明、富民、宾川3县（市）及四川省西南布拖县的硬枝野荞麦（Fagopyrum urophyllum)6个天然居群的遗传多样性和分化。硬枝野荞麦居群内维持有较高的遗传多样性,多态位点比率为50．0％,预期杂合度和观察杂合度分别为0．251和0．471。并对硬枝野荞麦（F.urophyllum)与栽培荞麦之间遗传变异作了比较。     

湖南新田野生大豆自然居群遗传多样性分析

利用SSR分子标记技术对8个来自湖南新田的野生大豆自然居群遗传多样性进行了分析.结果表明:(1)所分析的材料中73对SSR引物共检测到397个等位变异,等位变异数范围为2～10个,平均为5.4个;期望杂合度(He)的变化范围从0.16～0.82,平均为0.64.(2)分子方差分析发现,居群间存在着严重的遗传分化,群体69%的变异存在于居群间,31%的遗传变异存在于居群内.(3)新田的8个居群中桑梓路边(SZLB)和桑梓(SZ)两个居群的遗传多样性比其他群体的高,而新田1 km处(XT1)、新田2 km处(XT2)和新田6 km处(XT6)野生大豆居群的遗传多样性较低.(4)根据遗传距离可将8居群分为3类:新田1 km处和新田2 km处为一类;新田6 km处单独为一类;大冠岭上龙秀、龙秀后山、桑梓、桑梓路边和青龙等处为一类.(5)居群遗传距离和地理距离之间存在线性相关,相关系数为0.837(P<0.01);海拔与期望杂合度呈显著正相关,相关系数为0.92(P=0.001).研究结果表明,湖南新田野生大豆具有较高的遗传多样性,但不同居群的遗传多样性差异很大;位于该县高海拔山区大冠岭一带的野生大豆居群是湖南新田地区的一个多样性中心.