Molecular characterization of sweet potato(Ipomoea batatas[L.]Lam)germplasms for desirable traits by using simple sequence repeats markers

Every breeding program that aims to create new and improved cultivars with desired traits mostly relies on information related to genetic diversity.Therefore,molecular characterization of germplasms is important to obtain target cultivars with desirable traits.Sweet potato[Ipomoea batatas(L.)Lam]is widely considered the world’s most important crop,with great diversity in morphological and phenotypic traits.The genetic diversity of 20 sweet potato germplasms originating from Bangladesh,CIP,Philippines,Taiwan,and Malaysia were compared,which was accomplished by genetic diversity analysis by exploring 20 microsatellite DNA markers for germplasm characterization and utilization.This information was effective in differentiating or clustering the sweet potato genotypes.A total of 64 alleles were generated using the 20 primers throughout the 20 germplasm samples,with locus IBS97 having the highest number of alleles(5),whereas locus IbU33 had the fewest alleles(2).The alleles varied in size from 105(IbU31)to 213 base pairs(IBS34).The Polymorphism Information Content(PIC)values for the loci IbL46 and IBS97 varied from 0.445 to 0.730.IBS97 has the highest number of effective alleles(3.704),compared to an average of 2.520.The average Shannon’s diversity index(H)was 1.003,ranging from 0.673 in IbU3 to 1.432 in IBS97.The value of gene flow(Nm)varied between 0.000 and 0.005,with an average of 0.003,whereas genetic differentiation(FST-values)ranged between 0.901 and 1.000.The sweet potato germplasm included in this study had a broad genetic base.SP1 vs.SP9 and SP12 vs.SP18 germplasm pairings had the greatest genetic distance(GD=0.965),while SP1 vs.SP2 germplasm couples had the least genetic diversity(GD=0.093).Twenty genotypes were classified into two groups in the UPGMA dendrogram,with 16 genotypes classified as group“A”and the remaining four genotypes,SP10,SP18,SP19,and SP20,classified as group“B.”According to cluster analysis,the anticipated heterozygosity(gene diversity)of Nei(1973)was 0.591 on average.In summary,SSR markers successfully evaluated the genetic relationships among the sweet potato accessions used and generated a high level of polymorphism.The results of the present study will be useful for the management of germplasm,improvement of the current breeding strategies,and the release of new cultivars as varieties.

Genetic diversity of Oryza rufipogon Griff. in Hainan Province analyzed by ISSR and SSR markers

Assessment of genetic diversity is an essential component in germplasm characterization and conservation.There are three wild rice species in Hainan Province,including Oryza rufipogon Griff.In order to detect the genetic diversity of different populations of Oryza rufipogon in Hainan,ISSR(inter-simple sequence repeat)and SSR(simple sequence repeat)markers were used to investigate 180 accessions from six localities in Hainan.Fourteen ISSR primers amplified 185 alleles with 171(92.43%)polymorphic,the number of alleles ranged from 8 to 17,with an average of 13.14 alleles per locus.Thirty-eight pairs of SSR primers used in this study amplified 213 alleles with 190(89.20%)polymorphic,the number of alleles ranged from 2 to 14,with an average of 5.66 alleles per locus.Both ISSR and SSR analyses revealed a high level of genetic diversity in the wild populations.The population with the highest genetic diversity is Wanning(WN),and the population with lowest genetic diversity is Wenchang(WC).The results of a UPGMA cluster using the NTSYS program showed that each population has a low degree of genetic differentiation.Furthermore,the Mantel test revealed that the genetic similarities detected by ISSR and SSR were significantly correlated(r=0.8634,t=93.67)when detecting genetic diversity at the species level.The two molecular marker systems were able to determine the genetic diversity among Oryza rufipogon,and the two groups of indexes obtained by using the two markers have a high level of consistency.

Genetic diversity and population structure of Gossypium arboreum L. collected in China

Background: Gossypium arboreum is a diploid species cultivated in the Old World. It possesses favorable characters that are valuable for developing superior cotton cultivars.Method: A set of 197 Gossypium arboreum accessions were genotyped using 80 genome-wide SSR markers to establish patterns of the genetic diversity and population structure. These accessions were collected from three major G. arboreum growing areas in China. A total of 255 alleles across 80 markers were identified in the genetic diversity analysis.Results: Three subgroups were found using the population structure analysis, corresponding to the Yangtze River Valley, North China, and Southwest China zones of G.arboreum growing areas in China. Average genetic distance and Polymorphic information content value of G. arboreum population were 0.34 and 0.47, respectively, indicating high genetic diversity in the G. arboreum germplasm pool. The Phylogenetic analysis results concurred with the subgroups identified by Structure analysis with a few exceptions. Variations among and within three groups were observed to be 13.61% and 86.39%, respectively.Conclusion: The information regarding genetic diversity and population structure from this study is useful for genetic and genomic analysis and systematic utilization of economically important traits in G. arboreum.

Analysis of Genetic Polymorphic SSR Markers in Germplasm Resources of the Natural Colored Cotton

Short sequence repeats(microsatellite,SSR) and expressed sequence tags-SSR(EST-SSR) markers were employed to analyze the genetic diversity of natural colored cotton varieties.About

Genetic diversity and differentiation of fall webworm (Hyphantria cunea Drury) populations

The molecular variation and genetic relationships among five populations of the fall webworm （Hyphantria cunea Drury） in China were assessed using microsatellite technology. Ten microsatellite primers, producing polymorphic bands, were used across 300 samples. The percentage of polymorphic loci （PPB） was 98.36%; the percentage of polymorphic loci in five populations ranged from high to low in the following order： Cangzhou population, Yantai population, Qinhuangdao population, Dandong population, and Shijiazhuang population. The results showed that 34.38% of the total genetic variation of the fall webworm （GsT） occurs among populations, while most variation （65.62%） exists within populations. Nei＇s genetic distances ranged from 0.1386 to 0.3224. Using the unweighted pair group method with arithmetic mean （UPGMA）, Nei＇s genetic distances were analyzed by a clustering technique and the dendrogram shows that population differentiation is closely related to the time and geographic origin of the invasion. The major factors impacting genetic diversity of fall webworm populations are longitude, the plain area ratio, annual precipitation, latitude and time of invasion. The formation of genetic structure is correlated with characteristics of the life history and invasion ecology of the species.

Simple Sequence Repeat Analysis of Genetic Diversity in Primary Core Collection of Peach(Prunus persica)

In this study, the genetic diversity of 51 cultivars in the primary core collection of peach （Prunus persica （L.） Batsch） was evaluated by using simple sequence repeats （SSRs）. The phylogenetic relationships and the evolutionary history among different cultivars were determined on the basis of SSR data. Twenty-two polymorphic SSR primer pairs were selected, and a total of 111 alleles were identified in the 51 cultivars, with an average of 5 alleles per locus. According to traditional Chinese classification of peach cultivars, the 51 cultivars in the peach primary core collection belong to six variety groups. The SSR analysis revealed that the levels of the genetic diversity within each variety group were ranked as Sweet peach 〉 Crisp peach 〉 Flat peach 〉 Nectarine 〉 Honey Peach 〉 Yellow fleshed peach. The genetic diversity among the Chinese cultivars was higher than that among the introduced cultivars. Cluster analysis by the unweighted pair group method with arithmetic averaging （UPGMA） placed the 51 cultivars into five linkage clusters. Cultivar members from the same variety group were distributed in different UPGMA clusters and some members from different variety groups were placed under the same cluster. Different variety groups could not be differentiated in accordance with SSR markers. The SSR analysis revealed rich genetic diversity in the peach primary core collection, representative of genetic resources of peach.

Assessment of genetic diversity by simple sequence repeat markers among forty elite varieties in the germplasm for malting barley breeding

The genetic diversity and relationship among 40 elite barley varieties were analyzed based on simple sequence repeat （SSR） genotyping data. The amplified fragments from SSR primers were highly polymorphic in the badey accessions investigated. A total of 85 alleles were detected at 35 SSR loci, and allelic variations existed at 29 SSR loci. The allele number per locus ranged from 1 to 5 with an average of 2.4 alleles per locus detected from the 40 badey accessions. A cluster analysis based on the genetic similarity coefficients was conducted and the 40 varieties were classified into two groups. Seven malting barley varieties from China fell into the same subgroup. It was found that the genetic diversity within the Chinese malting barley varieties was narrower than that in other barley germplasm sources, suggesting the importance and feasibility of introducing elite genotypes from different origins for malting barley breeding in China.

基于EST-SSR标记的沙棘品种鉴定及指纹图谱构建

以沙棘(Hippophae rhamnoides)优良品种“实优1号”为材料,对其叶片进行转录组测序,利用微卫星识别软件(microsatellite identification tool,MISA)和Primer 3(version 2.3.4)对获得的序列进行简单重复序列(simple sequence repeat,SSR)位点挖掘和引物设计,以收集的42份沙棘品种为研究材料,开展聚合酶链式反应(PCR)和毛细管电泳检测,旨在开发一套多态性高、稳定性好和通用性强的表达序列标签微卫星(express sequence tags from simple sequence repeat,EST-SSR)引物,构建沙棘指纹图谱,从而实现沙棘品种的快速准确鉴定,并对沙棘品种间亲缘关系进行分析。“实优1号”转录组测序共获得6196个SSR位点,其中,重复基元类型为182种,SSR基序长度主要分布在10~21 bp区间,占全部SSR的81.58%,主要SSR重复类型为单核苷酸重复(48.72%)、二核苷酸重复(22.68%)和三核苷酸重复(18.85%)。利用筛选出的28对引物在42份沙棘品种中共检测出193个等位基因,等位基因数(Na)、有效等位基因数(Ne)、观测杂合度(Ho)、期望杂合度(He)、多态信息含量(PIC)和Shannon信息指数(I)等遗传多样性参数的均值分别为6.964、3.495、0.617、0.671、0.623和1.384。UPGMA聚类分析表明,42份沙棘品种间的遗传相似性系数为0.601~0.990,当遗传相似性系数为0.694时,供试品种可分为2组;当遗传相似性系数约为0.7402时,供试品种可分为3组。优选6对引物构建指纹图谱,可以实现沙棘品种的快速准确鉴定。该研究可为沙棘的良种鉴定、指纹图谱构建以及遗传多样性和亲缘关系分析等提供分子水平的理论基础和数据支撑。

Molecular Diversity and Association Analysis of Drought and Salt Tolerance in Gossypium hirsutum L. Germplasm

Association mapping is a useful tool for the detection of genes selected during plant domestication based on their linkage disequilibrium（LD）. This study was carried out to estimate genetic diversity, population structure and the extent of LD to develop an association framework in order to identify genetic variations associated with drought and salt tolerance traits. 106 microsatellite marker primer pairs were used in 323 Gossypium hirsutum germplasms which were grown in the drought shed and salt pond for evaluation. Polymorphism（PIC=0.53） was found, and three groups were detected（K=3） with the second likelihood ΔK using STRUCTURE software. LD decay rates were estimated to be 13-15 cM at r2 0.20. Significant associations between polymorphic markers and drought and salt tolerance traits were observed using the general linear model（GLM） and mixed linear model（MLM）（P 0.01）. The results also demonstrated that association mapping within the population structure as well as stratification existing in cotton germplasm resources could complement and enhance quantitative trait loci（QTLs） information for marker-assisted selection.

Grey correlational and SSR analyses of cotton hybrids

Ten upland cotton strains exhibiting 3 fiber quality traits and 8 yield traits, were grown for two years in an investigation of the correlation between grey relational analysis （GRA） and genetic identity in heterosis of cot- ton hybrid. The aim was to establish the optimal approach for heterosis prediction and parent selection. Plant traits data were collected and analyzed for GRA. In addition, 72 simple sequence repeat （SSR） markers were examined and 148 polymorphisms were detected. Correlation analysis of GRA, genetic identity, Ft fiber quality and yield heterosis was conducted. Significant differences were observed between the two analytic methods, whereas compa- rable predictions were given for yield heterosis. GRA for yield exhibited slightly higher correlation than genetic identity analysis, with a correlation coefficient of 0.49. GRA and genetic analysis exhibited overlapping yet dis- tinct advantages in heterosis prediction. Therefore, these analytical methods should be integrated to achieve the op- timal heterosis prediction and parent selection.

水芹SSR分子标记开发与遗传多样性分析

水芹是伞形科水芹属多年生草本植物,是一种重要的药食两用蔬菜作物。在中国,水芹的种植区域十分广泛,然而目前对其种质资源的鉴定、培育及遗传信息的研究较少。本研究利用溧阳白芹基因组开发水芹简单重复序列(SSR)分子标记,分析55份水芹的遗传多样性并用非加权组平均法(UPGMA)构建系统进化树,同时用SSR扩增条带数据构建DNA指纹图谱。结果显示,共鉴定到325699个SSR位点,其中单核苷酸SSR重复单元、二核苷酸SSR重复单元、三核苷酸SSR重复单元、四核苷酸SSR重复单元、五核苷酸SSR重复单元、六核苷酸SSR重复单元的出现频率分别为33.94%、54.62%、9.31%、1.66%、0.17%、0.29%,其中二核苷酸SSR重复单元数最多,有177887个,且A/T(占比为29.98%)和AT/AT(占比为35.70%)是较丰富的重复类型。UPGMA分析结果表明,33对高多态性引物[多态信息含量(PIC)>0.25]可将55份水芹材料分为4组。利用筛选出的4对引物(Oj-084、Oj-110、Oj-112、Oj-156)可以将55份水芹材料完全区分开,并且可构建指纹图谱。研究结果可为水芹种质资源鉴定、保护及分子遗传育种提供有力依据。

基于SSR标记的MCID法鉴定新疆地方梨品种及其遗传多样性分析

为深入探究新疆地方梨品种的遗传关系,本研究采用简单重复序列(SSR)标记技术对28个新疆地方梨品种进行遗传多样性分析。同时,利用人工绘制品种鉴别示意图(MCID)法建立品种鉴定图(CID)。结果表明,新疆地方梨品种间遗传差异较小,遗传分化为中等程度。通过构建新疆地方梨品种的聚类树状图,发现28个新疆地方梨具有较高的遗传多样性,遗传相似性系数为0.52~0.98。群体结构分析结果表明,最佳群体群组数值(K)=3时新疆地方梨品种被划分为3个组,各品种之间存在普遍的基因交流。采用MCID法构建新疆地方梨品种的CID图谱,并验证了该方法的有效性。本研究结果可为今后梨种质资源评价和利用提供理论依据。

SSR分子标记在玉米研究中的应用

SSR简单重复序列也称微卫星DNA,是由特异性的引物进行PCR扩增分析的一种分子标记技术。简要介绍了SSR分子标记的原理,分析了SSR分子标记在玉米的种质资源、品种纯度鉴定、真伪鉴定、遗传多样性、种质性状等方面的应用。通过研究表明:能够利用SSR分子标记技术对玉米的品种纯度鉴定、真伪性鉴定、遗传结构、亲缘关系、优劣群体的划分、种质性状等方面进行分析,同时也能为以后研究玉米的遗传连锁图谱构建、分子标记辅助育种、基因定位和种质资源等方面提供参考。

基于果形对草莓品种遗传多样性SSR分析

本研究利用SSR分子标记技术对国家果树种质北京草莓圃的173份草莓品种进行区分,根据果形利用PopGene软件对其做了聚类分析。结果显示,长圆锥形、短圆锥形、阔圆锥形、圆锥形/长楔形、圆锥形/平楔形、长楔形、近圆形、圆球形/卵圆形的观测纯合度都为0.846 2,扁球形、圆球形/半圆球形、短圆锥形/短楔形的观测纯合度都在0.923 1~0.948 7之间,但是短圆形纯合度最低,为0.692 3,由此发现与其他形状的亲缘关系较远,其余形状相互之间的亲缘关系较近。根据果形分析SSR扩增等位基因的多态性发现,173份草莓品种整体纯合体偏多,其中楔形/长圆锥形的无杂合体,而短圆形的纯合体最少,草莓品种整体亲缘关系较近。

Determination of the genetic diversity of vegetable soybean [Glycine max(L.) Merr.] using EST-SSR markers

The development of expressed sequence tag-derived simple sequence repeats(EST-SSRs) provided a useful tool for investigating plant genetic diversity.In the present study,22 polymorphic EST-SSRs from grain soybean were identified and used to assess the genetic diversity in 48 vegetable soybean accessions.Among the 22 EST-SSR loci,tri-nucleotides were the most abundant repeats,accounting for 50.00% of the total motifs.GAA was the most common motif among tri-nucleotide repeats,with a frequency of 18.18%.Polymorphic analysis identified a total of 71 alleles,with an average of 3.23 per locus.The polymorphism information content(PIC) values ranged from 0.144 to 0.630,with a mean of 0.386.Observed heterozygosity(H o) values varied from 0.0196 to 1.0000,with an average of 0.6092,while the expected heterozygosity(H e) values ranged from 0.1502 to 0.6840,with a mean value of 0.4616.Principal coordinate analysis and phylogenetic tree analysis indicated that the accessions could be assigned to different groups based to a large extent on their geographic distribution,and most accessions from China were clustered into the same groups.These results suggest that Chinese vegetable soybean accessions have a narrow genetic base.The results of this study indicate that EST-SSRs from grain soybean have high transferability to vegetable soybean,and that these new markers would be helpful in taxonomy,molecular breeding,and comparative mapping studies of vegetable soybean in the future.

分析中国栽培大豆遗传多样性所需SSR引物的数目

我国拥有极其丰富的大豆资源 ,传统的方法是根据农艺性状来分析其遗传变异 ,但农艺性状受自然环境和人为因素影响明显。随着大豆育成品种的增加 ,有限的表型变异已难以详细阐明我国 2万余份大豆品种的遗传变异情况 ,需要从DNA分子水平深入研究我国大豆资源遗传变异分布规律。本研究以 190份为大豆为初选核心种质的一个无偏样本 ,用 6 0对SSR引物扩增 ,获得 6 0 6个等位变异 ,平均每个位点有 10个等位变异 ,位点多态信息量范围从 0 5 5到 0 99,平均为 0 83。对 190份大豆相似系数矩阵的标准误分析表明 ,SSR引物数增加到 5 0左右时 ,再增加引物 ,标准误变化很小。共表型矩阵之间的相关性测验显示 ,当等位变异数达到 5 70以上 ,相互之间相关性极显著。从实验材料中选取东北春大豆类型作为一个小样本进行共表型矩阵相关性分析也有类似结果。用SSR方法分析中国栽培大豆 (G max)遗传变异关系时 ,只有等位变异数达到一定的范围时 ,才能真实地反映出品种之间的遗传变异关系。当群体的遗传变异范围变得相对较小时 ,分析个体之间的遗传变异关系所需的等位变异数目也相应降低。结合SSR位点在大豆基因组中的分布和基因多样性水平 ,能够找到分析栽培大豆遗传多样性的核心SSR引物。只有获得等位变异数在 5 70以上 ,

利用SSR标记研究85个玉米自交系的遗传多样性

利用均匀分布在玉米基因组上的70对SSR引物研究了73个国内外早熟类群玉米自交系、6份CIM-MYT标准测验种和6份国内标准测验种的遗传多样性。研究结果表明,70对引物在供试材料中共检测出286个等位基因变异,每对引物检测出2～8个等位基因,平均4.1个。每个位点的多态性信息量(PIC)变化为0.18～0.81,平均为0.58。85个自交系之间的遗传相似系数变化范围在0.43～0.93之间,平均为0.66。UPGMA聚类分析表明,85份供试自交系划分为6个亚群合并后为A、B两大类群,主坐标分析结果与聚类分析结果相似,均与自交系系谱来源关系基本一致。外来种质和未知自交系被划分到相应的杂种优势类群。

青藏高原东南缘老芒麦自然居群遗传多样性的SRAP和SSR分析

基于SRAP和SSR分子标记分析了青藏高原东南缘8个老芒麦自然居群遗传变异及群体遗传结构,获得下述结果:1)16对SRAP引物在90个单株中共扩增出384条可统计条带,其中多态性条带334条,占86.98%;16个SSR位点共检测出等位变异221个,平均每个位点13.8个,其中具有多态性的位点数192个,占86.88%。2)2种分子标记检测到老芒麦居群水平的基因多样性(He)分别为0.1092和0.1296,而物种水平的基因多样性达0.2434和0.3732。3)基于2种标记的Nei氏遗传分化指数Gst(0.5525和0.5158)表明老芒麦居群出现了较大程度的遗传分化,居群间的基因流非常有限,分别为0.4050和0.4694,老芒麦的遗传变异主要分布在居群间,居群内变异相对较小,Shannon多样性指数和分子方差变异(AMOVA)分析显示了相似的结果。4)基于聚类分析结果表明各居群间存在较为明显的地理分化,8个居群分化为采集地范围内的南、北和中部3个分支。通过对老芒麦遗传多样性和遗传结构的分析提出了对该物种遗传多样性的保护策略。

基于SSR标记和生物学性状进行桃遗传多样性的比较分析

利用生物学性状和SSR分子标记对50个桃品种进行遗传多样性研究,并将两种标记所得结果进行比较。14个生物学性状揭示出品种间的遗传相似系数范围为0～0.9,均值为0.4;而13对SSR引物揭示出遗传相似系数范围为0.52～0.97,均值为0.75。对两种标记进行共表型相关系数检测,结果显示两种结果无明显差异。经Mental检测,两种标记之间相关性较差,这可能与形成机理差异大有关。进行聚类和主成分分析,生物学性状方面品种主要分为3个组且与果肉性状相关,而分子方面品种主要分为2组,与地域相关。

茄子SSR遗传多样性及其农艺性状的关联分析

为分析茄子的遗传多样性,并挖掘与茄子农艺性状和品质性状相关的分子标记位点,选用18对高多态性SSR引物对70份国内外茄子材料进行多态性扫描、遗传多样性分析和群体结构分析。在此基础上采用Tassel 2.1 GLM方法进行标记位点与农艺性状和品质性状的关联分析。结果显示,18对SSR引物共检测到130个等位变异位点,平均每个引物7.2个等位变异。各种质遗传相似系数为0.56~0.93,平均0.68。基因多样性指数和Shan-non指数变幅分别为0.0700~0.4044和0.1036~0.5848。通过群体遗传结构分析将供试材料分为5个亚群。与农艺相关性状显著相关（P〈0.05）的位点有17个。供试材料之间存在一定的遗传差异,但遗传基础比较狭窄。