黔东南地方玉米品种SSR分子标记指纹图谱构建

利用SSR分子标记技术研究了黔东南地方玉米品种,筛选出48对引物,共检测出309个等位基因变异,每对引物检测出3～12个,平均为6.44个。每个位点的SSR多态信息量(PIC)变化为0.37～0.88,平均为0.70,标记索引系数平均为4.68。21个材料间的遗传相似性系数变化为0.57～0.90,平均为0.68。利用4对多态信息量高的引物建立了21个地方品种的DNA指纹图谱。     

Molecular diversity and population structure of the Ethiopian lentil (<Emphasis Type="Italic">Lens Culinaris</Emphasis> Medikus) genotype assessment using SSR markers

Knowledge of genetic diversity in germplasm is essential for formulating effective germplasm collection, conservation, utilization strategies in and crop improvement programs. It also provides an opportunity to take corrective steps infusing new genes to avoid risks associated with a narrow genetic bases. Genetic diversity analysis of 119 lentil genotypes of including 83 germplasm and 36 exotic genotypes from International Center for Agricultural Research in the Dry Areas was studied using 27 primers of simple sequence repeat (SSR) marker. Molecular analysis of variance showed variations of 82% within and 18% of the among population variance was explained. Degree of polymorphism observed among the populations was 100%. A total 122 alleles were detected, with 2 to 7 alleles per locus, with a mean of 4.52 alleles per locus. The estimated gene diversity value for 27 loci was 0.64. The average Shannon’s information index value of 1.19 was obtained showed the existence of high genetic variation within the genotypes. The genetic similarity indices ranged from 0.21 to 1.00. The SSR markers showed an average polymorphic information content (PIC) value of 0.58. Cluster analysis grouped the genotypes into five major clusters as distinct genetic populations. Diversity analyses revealed the existence of a high level of genetic variation among genotypes. This molecular diversity information provides a basis for future germplasm collection, utilization, and conservation strategies in gene banks and introducing exotic germplasm to widen the genetic base of the current lentil breeding population.     

利用SSR标记分析栽培种花生多态性及亲缘关系

利用11对SSR引物对24个花生栽培品种（包括四大类型）进行PCR扩增分析，其中4对检测到明显的多态性，共检测到33个等位基因变异，每一个位点上检测到的等位变异数为5～13个，平均为8.25个。根据扩增结果可以将24个品种中的21个相互区分。供试品种间的遗传相似系数值在0.2～1.0之间，平均为0.4788。根据UPGMA聚类分析结果,供试     

应用SSR和ISSR标记分析栽培香稻品种的遗传多样性

本研究利用24对SSR引物和36个ISSR引物,分析33份来源于亚洲10个国家的香稻品种的遗传多样性。分别获得93条和181条多态性片段,每个SSR座位可检测3～8个等位基因,平均为4．23个;每个ISSR引物可检测3～8个多态性位点,平均为5．03个。根据SSR和ISSR标记计算的品种间遗传相似系数分别在0．294～0．884之间和0．595～0．867之间。聚类分析表明,利用两种标记所得的聚类结果基本上一致,与品种所处的3种气候类型变化基本相符。进一步证实SSR和ISSR标记是研究水稻种质资源分类有效的工具。     

贵州省花生地方品种的遗传多样性

为进一步了解贵州花生地方品种的遗传多样性,合理、高效利用花生资源,用50对SSR引物评价了贵州不同地理来源的68份花生地方品种,结果多态性较好的41对引物共扩增出79个等位基因,平均每个引物1.93个;多态性信息量(PIC)变幅为0.045(PM43)~0.951(PM169);Shannon信息指数变幅为0.2518(PM79)~0.6926(PM188),平均0.5268;Nei遗传多样性指数变幅为0.1699(PM79)~0.4995(PM188),平均0.3556。采用类平均法对欧氏距离聚类分析表明,在遗传距离为0.94时将68个花生地方品种分成2个大类,同一地理来源、同一粒型的地方品种的亲缘关系并不是最近的,表明地方品种间亲缘关系与地理来源关系不大。说明贵州花生地方品种具有丰富遗传多样性。     

DNA profiling of pineapple cultivars in Japan discriminated by SSR markers

We developed 18 polymorphic simple sequence repeat (SSR) markers in pineapple (Ananas comosus) by using genomic libraries enriched for GA and CA motifs. The markers were used to genotype 31 pineapple accessions, including seven cultivars and 11 breeding lines from Okinawa Prefecture, 12 foreign accessions and one from a related species. These SSR loci were highly polymorphic: the 31 accessions contained three to seven alleles per locus, with an average of 4.1. The values of expected heterozygosity ranged from 0.09 to 0.76, with an average of 0.52. All 31 accessions could be successfully differentiated by the 18 SSR markers, with the exception of ‘N67-10’ and ‘Hawaiian Smooth Cayenne’. A single combination of three markers TsuAC004, TsuAC010 and TsuAC041, was enough to distinguish all accessions with one exception. A phenogram based on the SSR genotypes did not show any distinct groups, but it suggested that pineapples bred in Japan are genetically diversed. We reconfirmed the parentage of 14 pineapple accessions by comparing the SSR alleles at 17 SSR loci in each accession and its reported parents. The obtained information will contribute substantially to protecting plant breeders’ rights.     

大麦农艺性状与SSR标记的关联分析

为了解大麦亲本材料遗传特性和主要农艺性状特征,采用156份不同来源的大麦材料,在86个多态性SSR位点上检测遗传多样性,同时对7个农艺性状在两试验点作表型鉴定,利用GLM和MLM模型进行分子标记与表型性状的关联分析。结果共检测出392个等位变异,平均每个标记4.6个,PIC值变异范围为0.0612~0.8560。群体遗传结构分析将156份材料分为2个亚群。利用GLM模型分析结果表明,与株高、穗长、芒长、穗粒数和千粒重5个性状相关联的标记有18个,单个标记对表型变异的解释率为4.81%~20.75%;利用MLM模型分析,与株高、穗长、芒长、分蘖数、穗粒数和千粒重6个性状相关联的标记有14个,单个标记对表型变异的解释率范围为6.64%~31.55%。这些关联标记对后续研究有参考价值。     

利用SSR标记划分糯玉米的杂种优势群

利用SSR标记研究了30份糯玉米(Zea mays ceratinaKulesh)自交系的遗传变异。用21对扩增带型稳定的引物,从供试材料中检测出101个等位基因变异,每对引物检测等位基因2~10个,平均4.81个,平均多态性信息量0.60。经遗传多样性和聚类分析把供试糯玉米自交系划分为6个类群。     

陆地棉半野生种系的遗传多样性和亲缘关系分析

2000-2010年以陆地棉标准系TM-1和海岛棉标准系3-79作为对照,将陆地棉的7个半野生种系64份种质资源进行了简单重复序列(SSR)分子标记的遗传多样性和亲缘关系研究.112对SSR引物共检测出502个等位位点,其中多态性位点392个,约占总位点数的78%.每对引物扩增出2～10个等位位点,平均约为4.5个,其...     

Isolation and characterization of 28 polymorphic SSR loci from castor bean (<Emphasis Type="Italic">Ricinus communis</Emphasis> L.)

Castor bean (Ricinus communis) is cultivated for seed oil throughout tropical and subtropical regions but the understanding of its genetic variability is limited. Because applicable microsatellite markers are not sufficient, we isolated and characterized polymorphic simple sequence repeat (SSR) loci acquired from a microsatellite-enriched genomic DNA library of castor bean. Finally, 28 SSR loci revealed polymorphisms in a castor bean collection consisting of 72 accessions. A total of 73 alleles were detected, with an average of 3.18 alleles per locus, and the polymorphism information content (PIC) ranged from 0.03 to 0.47 (mean = 0.26). Values for observed (H_O) and expected (H_E) heterozygosity ranged from 0.00 to 0.19 (mean = 0.11) and from 0.04 to 0.54 (mean = 0.31), respectively. To understand genetic relationships within the castor bean collection, a dendrogram was constructed based on profiles of the 28 SSR loci. These newly developed SSRs will be useful tools for assessing genetic diversity and population structure in castor bean.     

Identification of SSR markers which could differentiate blast disease resistance accessions in finger millet (<Emphasis Type="Italic">Eleusine coracana</Emphasis> (L.) Gaertn.)

Microsatellites also known as SSR are the class of repetitive DNA sequences present throughout the genome of all eukaryotic organisms. The present study identified SSRs using biotinylated beads capture method. Ten sets of primers were designed based on sequences having at least (CT)10 repeats. A total of 27 accessions having a mix of both African (resistant) and Indian origin (resistant and susceptible) were assessed using 30 microsatellite markers. Amplification products were obtained for all 30 primers studied; 25 out of these primers were found to be polymorphic with 13 primers showing two alleles per locus. The current study identified markers which could differentiate between resistant and susceptible accessions and also segregate accessions based on geographical region. These informative SSR markers can be used in finger millet genetic improvement projects.     

基于SSR标记的雪茄烟种质资源指纹图谱库的构建及遗传多样性分析

为从分子水平研究我国雪茄烟种质资源的遗传多样性差异并建立雪茄烟品种的DNA指纹图谱数据库,本研究利用43对多态性好的SSR引物对220份雪茄烟种质进行遗传多样性分析,筛选出14对核心引物对雪茄烟种质进行指纹图谱的构建。结果表明,43对SSR引物在220份雪茄烟种质材料中共扩增出243个等位基因,平均每个标记5.65个,变幅为2~13,每个位点的多态性信息量(polymorphism information content,PIC)变化为0.2078~0.9087,平均为0.6360。有效等位基因数(number of effective alleles,N_e)范围为1.3081~11.7876,平均有效等位基因数为3.9077;观测杂合度(observed heterozygosity,H_o)变化范围为0.0828~0.7639,平均为0.3191;预期杂合度(expected heterozygosity,H_e)的变化范围为0.2361~0.9172,平均为0.6809;种群平均Shannon遗传多样性指数(Shannon genetic diversity index,I)为1.3756,遗传距离在0.0233~0.9286之间,平均遗传距离0.6816。聚类分析表明,在遗传距离为0.74处,可将供试雪茄烟资源分为3个类群。Structure群体遗传结构分析和主成分分析将所有的供试材料划分为2个类群。根据引物的分析和表型鉴定结果,确定良种、辅善和满耳朵,山东大叶和牡丹江05-1,Florida513和CA0701为异名同种,一个品种保留一份种质,剩余216份不同种质。从43对SSR引物中筛选出14对可区分所有供试材料的SSR引物作为核心引物构建了216个雪茄烟品种的指纹图谱。我国雪茄烟种质资源具有较高水平的遗传多样性,本研究构建的雪茄烟种质资源SSR指纹图谱库及遗传分析的结果在分子水平上为筛选、鉴定优质雪茄烟种质资源、挖掘重要基因以及拓宽雪茄烟遗传育种基础等工作提供科学依据。     

Identification of QTLs for agronomic traits using association mapping in lentil

A diverse panel of 96 genotypes of lentil was used in this study to identify QTL for nine agronomic traits through marker-trait association analysis. This study showed significant genetic variability among the lentil genotypes for nine agronomic traits and had medium to large broad sense heritability estimates (h² =?0.58–0.95). Screening of 534 SSR markers resulted in 266 polymorphic loci that generated 697 alleles ranging from 2 to 16 alleles per locus across the genotypes. The model-based population structure analysis identified two distinct subpopulations among lentil genotypes and each subpopulation did not show any admixture. Marker trait association (MTA) analysis following ML model resulted in the identification of 24 MTAs for nine traits at P?<?0.01. The per cent of phenotypic variation explained by each associated marker with particular agronomic trait ranged from 7.3 to 25.8%. The highest proportion of total phenotypic variation (23.1–25.8%) was explained by the QTLs controlling the primary branches/per plant. In the present study, few EST-SSR markers showed significant association with days to maturity, pods/plant, secondary branches/plant, 100 seed weight, yield/plant and reproductive duration and explained large phenotypic variation (7.3–23.8%). Hence, these markers can be used as functional markers in lentil breeding program for developing improved cultivars.     

新疆加工番茄品种遗传多样性的SSR分析

选用已筛选出带型稳定的44对SSR引物,对新疆主栽以及美国引进的加工番茄品种进行遗传多样性分析,结果表明：在24个品种间共扩增了283个等位基因,每对引物的等位基因数变化范围在2~17之间,平均为4.422个;有效等位基因为189.638,平均为4.310;每个SSR位点的多态性信息量变化范围为0.140~0.915,平均为0.662;基因型多样性变化范围为0.325~2.620,平均值为1.438;24个品种间的遗传相似系数变幅为0.715~0.924,平均值为0.820,且95%的供试品种其遗传相似系数在0.715~0.800之间,亲缘关系较近。以遗传相似系数为原始数据,按UPGMA方法将供试品种划分为4大类群,结合系谱分析结果表明,新疆的加工番茄品种遗传多样不够丰富,多数品种间的亲缘关系较近,欲进一步提高新疆加工番茄的产量及品质性状还需拓宽亲本选择范围,扩大遗传背景。     

Assessment of genotypic variation among cultivated durum wheat based on EST-SSRS and genomic SSRS

Genotypic variation of 64 durum lines, landraces, and varieties wereinvestigated using three sources of microsatellites (SSRs). 245 primer pairswere used to detect polymorphism in this collection. A total of 42polymorphic SSR primer pairs consisting of 22 EST-SSR, 11 XGWM and9 WMC were used for genotyping. The EST-SSRs primers produced highquality markers, but were least polymorphic (25%) compared to 53%for the other two sources. The 42 primers detected 189 polymorphicalleles with an average of 4.5 alleles per locus. The coefficient of similaritybetween accessions ranged from 0.28 to 0.70. The estimate of similaritydiffered when each source of SSRs was analyzed independently. This studyshowed that SSRs are highly polymorphic in durum. The data presentedhere provides a platform to develop a genotypic database for durum wheatthat will facilitate the exploitation of its genetic resources.     

普通豇豆应用核心种质的SSR指纹图谱构建及多样性分析

不同种质资源群体的多样性及遗传背景分析可为种质资源收集保存及创新利用提供有效信息。对不同来源的88份普通豇豆应用核心种质资源进行了14个SSR位点的指纹图谱构建及多样性分析,共检测到39个等位变异,每对引物检测到2~5个,平均为2.79。多态信息含量（PIC）变幅为0.25~0.72,平均为0.58。UPGMA聚类结果显示,除了3对种质外,39个等位变异可将其他材料有效区分,且在遗传相似系数为0.63时,88份种质可分为4个类群,地理来源相同的材料有聚在一起的趋势。     

Identification and characterization of EST-SSRs in finger millet (<Emphasis Type="Italic">Eleusine coracana</Emphasis> (L.) Gaertn.)

In recent years, microsatellites have become the most used markers for studying population genetic diversity. The increased availability of the DNA sequences has given the possibility to develop EST-derived SSR markers. A total of 1,927 ESTs of Eleusine coracana available in the NCBI database were mined for SSRs. Di-nucleotides are the most occurring motifs accounting for more than 50% of the repeats, of which GA was the most abundant motif and tetra-nucleotides are the least occurring motifs. Of the 132 markers identified, 30 primer pairs based were synthesized. SSR markers were used for variety discrimination and genetic assessment in 15 finger millet accessions; 20 primers showed polymorphism and 13 primers were identified as having a PIC value above 0.5. On the basis of the distribution of these polymorphic alleles, the 15 accessions were classified into two groups. This study has demonstrated the potential of EST-derived SSR primer pairs in finger millet. These primers will serve as valuable source for further breeding programs.     

Analysis of molecular variance and population structure in southern Indian finger millet genotypes using three different molecular markers

The genetic relationship among 42 genotypes of finger millet collected from different geographical regions of southern India was investigated using random amplified polymorphic DNA (RAPD), inter simple sequence repeats (ISSR), and simple sequence repeats (SSR) markers. Ten RAPD primers produced 111 polymorphic bands. Five ISSR primers produced a total of 61 bands. Of these, 23 bands were polymorphic. The RAPD and ISSR fingerprints revealed 71.3 and 37.4% polymorphic banding patterns, respectively. Thirty-six SSR primers yielded 83 scorable alleles in which 62 were found to be polymorphic. Out of 36 SSR primers used, 14 primers (46.6%) produced polymorphic bands. The SSR primer UGEP7 produced a maximum number of six alleles. Mean polymorphic information content (PIC) of RAPD, ISSR and SSR were 0.44, 0.28, and 0.14, respectively. Molecular variances among the population were 2, 11, and 1% for RAPD, ISSR, and SSR markers, respectively. SSR produced 99% molecular variance within individuals. RAPD and ISSR markers produced a low level of molecular variance within individuals. The STRUCTURE (model-based program) analysis revealed that the 42 finger millet genotypes could be divided into a maximum of four subpopulations. Based on the Bayesian statistics, each RAPD and SSR marker produced three subpopulations (K=3), while ISSR marker showed four subpopulations (K=4). This study revealed that RAPD and SSR markers could narrow down the analysis of population structure and it may form the basis for finger millet breeding and improvement programs in the future.     

亚洲棉种质资源的SSR遗传多样性分析

 对我国棉花中期库保存的200份不同地理来源的亚洲棉代表性样本进行了SSR遗传多样性分析,结果表明：亚洲棉分子水平的遗传多样性较高。83个多态性位点共检测到368个等位基因变异,其中多态性的等位基因数为329个,平均每个SSR位点3.964个。位点多态性信息量（PIC）变幅为0.010～0.882,平均0.578,PIC值大于0.7的标记有33个（占39.8%）。基因多样性（H′）变幅为0.031～2.163,有效等位基因数（Ne）变幅为1.010～8.496。华南棉区基因遗传多样性最高,其次为长江流域棉区、黄河流域棉区,从理论上支持被广泛接受的亚洲棉在我国的传播路线是由南到北,华南棉区是中棉种系的遗传多样性富集中心。利用软件NYSTS-pc2.20,采用类平均法(UPGMA)进行聚类分析,种质间SSR相似系数变幅为0.58～0.997,平均0.745,在阈值0.73处200份亚洲棉聚为8个类群,贵池小子棉白子单独聚为一群,与其他种质遗传距离较远。遗传距离和地理距离没有必然联系,但种质间亲缘关系处于极端远或极端近时,则地理距离一般也趋于较远或较近。     

甘蔗栽培种单倍体基因组SSR位点的发掘与应用

甘蔗是世界上最重要的糖料作物之一,由于尚未完全破译栽培种基因组,导致SSR标记匮乏,难以覆盖全基因组,限制了甘蔗遗传研究的进展。本研究以栽培种R570的4660个BAC文库片段序列(累计总长为382 Mb,预测到25,316个编码蛋白基因)组装成的一套甘蔗单倍体基因组的模板,利用MISA (Microsatellite identification tool)软件,发掘SSR位点;并综合分析其与4种禾本科植物(高粱、玉米、水稻和二岁短柄草)SSR位点的分布特征;选取50对以TG和AG重复基序的SSR引物,分别利用4个甘蔗属材料(R570、ROC1、LA purple和SES208)和24个重要甘蔗亲本,对SSR引物进行扩增效率验证和多态性分析。共发掘到27,241个SSR位点,平均每个BAC片段有6.29个SSR位点,平均密度为71.33个SSR Mb?1,远低于高粱的平均密度(350.00个SSR Mb?1)。在重复基序中,占比前2位的分别为单核苷酸基序(11,079个)和三核苷酸重复基序(6447个),合计占总SSR位点数的64.33%。与甘蔗不同的是, 4种禾本科植物中的三核苷酸基序类型数量最多、占比最大。此外,在单核苷酸重复基序中, A/T所占比例最高,为84.8%, C/G所占比例最低,为15.2%;在三核苷酸重复基序中, TGT/ACA所占比例最高,为16.04%。总之,禾本科植物基因组富含A/T的基序。在50对SSR引物(TG基序41对和AG基序9对)的多态性验证中,共有45对(90%)能够扩增出清晰的条带,其中35对(70%)在4个甘蔗材料上呈现多态性。进一步利用20对多态性较高的SSR引物对24个甘蔗重要亲本材料进行分析,共扩增到95个等位基因,平均每对引物扩增4.75个,验证了这些引物应用于甘蔗遗传多样性研究的可行性。本研究鉴定的甘蔗栽培种单倍体基因组SSR标记,有效增加了甘蔗遗传研究中可用的分子标记数量,可直接用于甘蔗群体遗传多样性分析和重要性状遗传机制的解析,为甘蔗分子育种的深入研究奠定了基础。