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1.
随着生物科技的进步,大量的植物表达序列标签（expressed sequence tags,ESTs）已经成为开发SSR标记的重要资源。EST-SSR作为一种新型分子标记,其多态性可能与基因功能直接相关,而且在相近植物间具有良好通用性,使得EST-SSR标记在实际应用中更具价值。采用计算机方法大规模发掘EST-SSR多态性位点极大地提高了SSR标记开发效率。尤其随着新一代测序技术的成熟以及测序成本的急剧下降,利用新一代测序技术产生大量的转录组数据进行EST-SSR多态性标记的计算机大规模发掘将对SSR标记的开发带来深远影响。本文简要介绍了植物EST-SSR分布特点,并对EST-SSR标记开发现状以及相关应用作了综合评述,此外,还对EST-SSR标记的计算机开发新策略进行了展望。  相似文献   

2.
为了明确菠菜EST序列中SSR的总体特点,开发菠菜EST-SSR引物;为利用EST-SSR引物进行菠菜性别相关特异序列的克隆奠定基础,本文从NCBI上获得1093条EST,利用在线软件SSRIT检测所含SSR序列,并进行分析。共检索出68条SSR序列,分布于64条EST中,检出率为6.22%,包括22种重复基元。其中二核苷酸重复基元的EST-SSR占主导地位,占总SSR数目的32.3%。利用在线引物设计软件Primer3.0设计了7对EST-SSR引物,在适合的PCR反应体系下,分别以雌、雄菠菜DNA基因组为模板,对设计的EST-SSR引物进行筛选,结果显示以EST序列HS097148设计的一对引物从菠菜雌雄基因组中扩增出一条雄性特异的条带,表明通过菠菜EST-SSR引物获得菠菜性别相关特异序列是可行的。  相似文献   

3.
杨旭  杨志玲  谭美  程小燕 《核农学报》2019,33(7):1318-1329
为探明厚朴的基因组信息,开发合适的生物学工具以促进厚朴的遗传育种,本研究利用Illumina-Solexa测序技术对厚朴根、茎皮、叶不同组织的9个样本进行转录组分析,获得大量的序列信息,并以转录组序列为基础进行厚朴SSR分子标记的大规模发掘。结果表明,共获得了109 526条厚朴转录组序列,平均序列长度为1 023 bp,通过与蛋白数据库NR、Swiss-Prot、GO、KOG和KEGG五个数据库比对,分别有6 0361、39 942、65 535、51 351和27 310条厚朴转录组的Unigene被注释;厚朴转录组表达信息进行大通量SSR位点的发掘,发现了含SSR位点的序列25 925条,共27 721个SSR位点,SSR出现的频率为23.67%。厚朴转录组共发现了374种碱基重复模式,且CT/GA和AG/TC是主要的双碱基重复序列,AGA/TCT和AAG/TTC是主要的三碱基重复序列;随机挑选的180对SSR引物中有104对产生清晰可重复的条带,21对引物具有多态性,厚朴SSR引物多态性较高。本研究结果为厚朴及近缘种的遗传多样性分析、遗传图谱构建及比较基因组研究奠定了一定的理论基础。  相似文献   

4.
The development of EST-SSR in foxtail millet (Setaria italica) for polymorphism and transferability study was reported here. From 1213 EST sequences, 30 SSRs were obtained and primers were designed for 26 SSRs. Among them, four pairs of SSR primers amplified polymorphic products in 12 foxtail millet cultivars and one accession of Setaria viridis, a wild relative of foxtail millet, with 10 alleles detected for the four loci and 2.5 alleles per locus. In addition, ten SSR markers could be transferred to other nine Gramineae species. The putative functions of 11 ESTs containing polymorphic and transferable SSRs were also identified.  相似文献   

5.
Breeding progress in black raspberry (Rubus occidentalis L.) has been limited by a lack of genetic diversity in elite germplasm. Black raspberry cultivars have been noted for showing very few phenotypic differences and seedlings from crosses between cultivars for a lack of segregation for important traits. Despite these challenges, little molecular work has been done to explore genetic diversity and relationships in wild and cultivated black raspberry germplasm. Microsatellite, or simple sequence repeat (SSR), markers are highly polymorphic codominant markers useful for studying genetic diversity, population genetics, genetic fingerprinting and other applications. We examined genetic diversity in 148 wild and cultivated black raspberry accessions using 21 polymorphic SSR markers. Black raspberry cultivars clustered tightly and showed higher than expected heterozygosity while that of wild accessions was low. Relationships between wild black raspberry accessions were poorly resolved and regional clusters were mostly absent from our analysis. Our results indicated that wild black raspberry germplasm is a relatively untapped resource available for future breeding.  相似文献   

6.
Genetic diversity of 54 accessions of Aegilops tauschii from five countries was assessed using sequence-tagged microsatellites (or simple sequence repeats, SSRs) and amplified fragment length polymorphisms (AFLPs). In the case of AFLP analysis, a total of 256 amplification products obtained, 234 of them were polymorphic across all the 54 accessions. A total of 224 fragments were obtained from the 24 SSR primers and 219 of fragments were polymorphic across all the genotypes screened. Based on both AFLP and SSR markers, the highest percentage of polymorphisms were obtained in Iranian and accessions of unknown origin. The highest polymorphic information content (PIC) value was observed for SSRs (0.82) while the highest marker index (MI) value was for AFLPs (8.5) reflecting the hyper-variability of the first and the distinctive nature of the second system. Principal co-ordinate analysis (PCO) revealed congruent patterns of genetic relationships for both data sets, but did not group accessions strictly according to their geographical origins. Poor correlation was found between AFLP and SSR marker loci. This low association may be due to low number of AFLP and SSR markers. These results show that molecular markers can help to organize the genetic variability and expose useful diversity for breeding purposes.  相似文献   

7.
The origin of cultivated potatoes has remaining questions. In this study, 237 accessions of all cultivated species and 155 accessions of wild species closely related to cultivated potatoes, including their putative ancestors, were analyzed using 15 plastid microsatellites (SSRs) to investigate genetic diversity and their relationships with the wild species. We here used polymorphic plastid SSRs we developed from potato plastid genome sequences as well as already known plastid SSR markers. All 15 loci were polymorphic and identified a total of 127 haplotypes. Dramatic decreases in levels of genetic diversity were revealed in landraces in comparison with wild ancestor species. The plastid SSR results showed a decrease in haplotype number and diversity from Peru to both north and south. Phylogenetic analysis revealed two distinct groups. One of them, group A, contained the majority of accessions of cultivated species of the Solanum tuberosum Andigenum group including all accessions of cultivated diploid and triploid cytotypes of this group (S. chaucha, S. phureja, and S. stenotomum by a former taxonomic system) and most of tetraploid accessions of the S. tuberosum Andigenum group (S. tuberosum subsp. andigenum), and the majority of accessions of wild ancestors from the northern members of the S. brevicaule complex. Another group B comprised most of the wild species accessions and almost exclusively hybrid cultivated species which have introgressed plastid genomes from the other wild gene pools. Lack of clustering of traditional cultivated species (as used above) support a revised group classification of S. tuberosum.  相似文献   

8.
Lolium and Festuca are two important genera of cool-season forage and turf grasses worldwide. Lolium temulentum L. (darnel ryegrass) has been proposed as a model species for genomics studies of cool-season forage and turf grasses. A study with 41 darnel ryegrass, three tall fescue (Festuca arundinacea Schreb.), two tetraploid fescue (F. glaucescens), and two meadow fescue (F. pratensis) genotypes was initiated to (i) identify a set of microsatellite (simple sequence repeats) markers useful for L. temulentum L., and (ii) to utilize such markers for assessing the genetic variability of L. temulentum accessions collected from different geographical regions of the world. A total of 40 tall fescue (TF) EST-SSRs and 60 Festuca–Lolium (F × L) genomic SSRs were screened on a subset of eight genotypes. The selected 30 tall fescue EST-SSRs and 32 F × L genomic SSRs were used for further analysis of genotypes. The TF-EST- and the F × L genomic-SSRs identified 10.3 and 9.3 alleles per marker, respectively with an average polymorphic information content (PIC) value of 0.66. The phenogram based on 319 EST-SSR and 296 genomic SSR fragments, grouped L. temulentum accessions into three major clusters except for accession ABY-BA 8892.78. Lolium temulentum accession ABY-BA 8892.78 did not cluster with any other accession. The Festuca clusters were distantly related with darnel ryegrass clusters with a similarity coefficient of 0.26. The selected set of tall fescue EST- and F × L genomic SSRs were useful in assessing L. temulentum genetic diversity and could benefit the genetic improvement of members of the Festuca–Lolium complex.  相似文献   

9.
A barley core collection can be studied extensively and the derived information can be used to identify loci/genes for the genetic improvement of quantitative and qualitative traits. To assess genetic diversity, allelic variation and population structure of Egyptian barley, 134 barley genotypes collected from a different region along with 19 cultivated genotypes obtained from of the Egyptian Agricultural Research Center. All genotypes were analyzed with 261 polymorphic SSR and SNP alleles. The mean number of alleles per locus was 4 and PIC was 0.49, while the level of genetic diversity was 0.55 ranging from 0.03 to 0.82. The genotypes were assigned to three subpopulations that were consistent with their origins. The genetic variation within population was higher (51%) than among population at the molecular levels (FST =?0.491 when P?<?0.10). The level of polymorphic variation was highest in subpopulations-II, due to collected from different regions with different ear-types thus, expected to contain more diversity than local genotypes in subpopulations-I and subpopulations-III. The structured study found that the 153 barley genotypes are in harmony with clustering approaches using the SSR and SNP genotypic data in a neighbor-joining tree and principal components analysis, which identified three subpopulations. These results demonstrated genetic diversity among the Egyptian barley genotypes can be applied to suggest approaches, such as association analysis, classical mapping population development, and parental line selection in breeding programs. Therefore, it is necessary to use the exotic genotypes as the genetic resources for developing new barley cultivars in Egypt.  相似文献   

10.
Chawali and Lekali are two common farmer’s barley varieties or landraces in Jumla, Nepal (2,240–3,000 m) with small to bold grains and wide adaptation from irrigated low lands to high hills. This study was undertaken to test whether features of the traditional seed system can significantly influence the diversity of a crop and its conservation on-farm. In Jumla (high-hill), the barley seed system is completely informal and is mainly from farmer to farmer. In the present study, the seed flows and the pattern of genetic diversity in barley were investigated to detect differences between the two varieties and test the divergence among populations of each variety These data suggested that Chawali, the more common variety, was less subject to homogenising gene flow between farms than was Lekali. A total of 128 farming households were surveyed for seed supply information and 128 populations for each landrace from two villages: Kartikswami and Talium were collected for SSR diversity analysis. Some 92 SSRs were screened in an initial sample of 20 barley populations of both landraces and 2 improved varieties (LG-51 and Soluwa). Of the 81 SSRs that consistently amplified, only 15 SSRs (19%) were polymorphic with gene diversity values ranging from 0.09 to 0.71. A medium to low diversity was detected among the landrace populations of barley varieties. Chawali populations were less polymorphic within ecological groups, and more divergent between than were Lekail populations. This result accords with Chawali having a more conservative local seed system.  相似文献   

11.
Eight genomic SSR markers with a M13 tail attached were used to assess the genetic diversity of 72 Ussurian Pear accessions (Pyrus ussuriensis Maxim.) in China. The M13-tailed method was effective in discriminating all the 32 wild accessions. All the 40 Ussurian Pear cultivars could be successfully discriminated with the exception of 4 sets of synonymies or spots. A total of 108 alleles were obtained with an average of 13.5 per locus. The expected heterozygosity, observed heterozygosity, and power of discrimination were 0.78, 0.63, and 0.86 respectively. Three triploid cultivars (‘Anli’, ‘Ruan’er’, and ‘Pitaiguo’), and one wild accession, P. ussuriensis ‘Xilin-3’, showed three alleles at some SSRs. The number of alleles and observed heterozygosity per locus for 40 Ussurian Pear cultivars were 9.1 and 0.62, respectively, lower than the values of 32 wild accessions which were 11.3 and 0.65, respectively. A dendrogram based on the SSR genotypes was obtained, showing two major groups corresponding to cultivated group and wild group. All the cultivars fell into the cultivated group. Some subgroups (Nanguoli subgroup, Zhibazi subgroup, Xiangshuili subgroup, Balixiang subgroup, Anli subgroup) could be found in the cultivated group. A very close relationship between ‘Huagaili’ and ‘Miansuan’, and a close relationship between ‘Anli’ and a wild accession, P. ussuriensis ‘Huangshanli’ could be found in Anli subgroup. ‘Nanguoli’ and ‘Xiaowuxiang’ showed a close relationship with at least one identical allele at each locus with the exception of NH015a.  相似文献   

12.
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。  相似文献   

13.
A total of 8117 suitable SSR-contaning ESTs were acquired by screening from a Malus EST database, among which dinudeotide SSRs were the most abundant repeat motif, within which, CT/TC followed by AG/GA were predominant. Based on the suitable sequences, we developed 147 SSR primer pairs, of which 94 pairs gave amplifications within the expected size range while 65 pairs were found to be polymorphic after a preliminary test. Eighteen primer pairs selected randomly were further used to assess genetic relationship among 20 Malus species or cultivars. As a result, these primers displayed high level of polymorphism with a mean of 6.94 alleles per locus and UPGMA cluster analysis grouped twenty Malus accessions into five groups at the similarity level of 0.6800 that were largely congruent to the traditional taxonomy. Subsequently, all of the 94 primer pairs were tested on four accessions of Pyrus to evaluate the transferability of the markers, and 40 of 72 functional SSRs produced polymorphic amplicons from which 8 SSR loci selected randomly were employed to analyze genetic diversity and relationship among a collection of Pyrus. The 8 primer pairs produced expected bands with the similar size in apples with an average of 7.375 alleles per locus. The observed heterozygosity of different loci ranged from 0.29 (MES96) to 0.83 (MES138), with a mean of 0.55 which is lower than 0.63 reported in genome-derived SSR marker analysis in Pyrus. The UPGMA dendrogram was similar to the previous results obtained by using RAPD and AFLP markers. Our results showed that these EST-SSR markers displayed reliable amplification and considerable polymorphism in both Malus and Pyrus, and will contribute to the knowledge of genetic study of Malus and genetically closed genera.  相似文献   

14.
Kunzea pomifera F.Muell. (muntries) is a native food crop in Australia that produces spicy apple-flavoured berries. Orchards were established in the 1990s using a variety of wild-collected selections. No published records are currently available of formal plant breeding although the Rivoli Bay selection is the most widely planted. DNA markers are needed to reliably assess genetic diversity and validate plant selections, however genomic resources of K. pomifera are not available. Initially, based on the large MYB gene family, eight MYB gene fragments were amplified and cloned using a degenerate PCR primer strategy. Targeted amplicon sequencing was subsequently used to evaluate diversity of 19 K. pomifera plants and this revealed the presence of single nucleotide polymorphisms and simple sequence repeats (SSRs). Three genic SSR markers were developed from K. pomifera MYB gene fragments and another three genic SSRs were successfully transferred from K. pulchella (Lindl.) A. S. George. A high transfer rate (75%) was obtained due to selecting SSRs in genic regions (introns or intergenic regions) and using Eucalyptus grandis W.Hill sequences to identify conserved regions. The six SSRs were highly polymorphic with observed heterozygosity (Ho) (average Ho = 0.63) and polymorphic information content (PIC) (average PIC = 0.54). Using the six genic SSRs, 15 unique genotypes were identified from the K. pomifera collections in two orchards. Importantly, plants in a new diversity panel were genotyped and suspected mis-labeling events confirmed. The genic SSRs and approach developed will benefit marker assisted selection programs by increasing the proportion of genic markers that could be linked to important horticultural traits.  相似文献   

15.
The aim of this study was to identify the group of highly polymorphic microsatellite markers for the identification of six pear cultivars (P. communis) and two individuals of wild pear (P. pyraster). From among 40 tested SSR markers, 19 were selected to profile genetic diversity in pear genotypes due to high polymorphisms. These markers showed high heterozygosity levels (0.5–1) and, on average, 6.4 alleles per marker were found. The set of microsatellite markers employed in this study demonstrated usefulness of microsatellite markers for the identification of pear genotypes. The examined wild forms were represented in this study by only two individuals of P. pyraster. It can be assumed that these forms were distinctly different from the cultivated pear cultivars.  相似文献   

16.
Thirty-four durum wheat cultivars representing the Tunisian durum (Triticum durum Desf.) wheat collection and seven wild species of wheat relatives (Triticum turgidum L., T. dicoccon Schrank., T. dicoccoides (Körn) Schweinf., T. araraticum Jakubz., T. monococcum L., Aegilops geniculata Roth, and Aegilops ventricosa Tausch) were analysed with amplified fragment length polymorphism (AFLP) and microsatellite (SSR) markers. Both marker systems used were able to differentiate durum wheat cultivars from the wild relatives and to specifically fingerprint each of the genotypes studied. However, the two marker systems differed in the amount of detected polymorphisms. The 15 SSR markers were highly polymorphic across all the genotypes. The total number of amplified fragments was 156 and the number of alleles per locus ranged from 3 to 24 with an average of 10.4. Two SSR markers alone, Xwms47 and Xwms268, were sufficient to distinguish all 34 durum wheat genotypes. The five AFLP primer pair combinations analysed yielded a total of 293 bands, of which 31% were polymorphic. The highest polymorphic information content (PIC) value was observed for SSRs (0.68) while the highest marker index (MI) value was for AFLPs (7.16) reflecting the hypervariability of the first and the distinctive nature of the second system. For durum wheat cultivars, the genetic similarity values varied between 31.3 and 81% for AFLPs (with an average of 54.2%), and between 3.6 and 72.7% for SSRs (with an average of 19.9%). The rank correlation between the two marker systems was moderate, with r = 0.57, but highly significant. Based on SSR markers, highest genetic similarity (GS) values were observed within the modern cultivars (37.3%), while the old cultivars showed a low level of GS (19.9%). Moreover, the modern cultivars showed low PIC and MI values. UPGMA Cluster analysis based on the combined AFLP and SSR data separated the wild wheat species from the durum wheat cultivars. The modern cultivars were separated from the old cultivars and form a distinct group.  相似文献   

17.
航恢七号空间诱变变异株系的稻瘟病抗性研究   总被引：2，自引：0，他引：2  
本研究对卫星搭载水稻品种航恢七号SP3代株系进行稻瘟病抗性鉴定研究,并对抗性变异株系作分子生物学分析。结果表明,250个航恢七号SP3代农艺经济性状优良株系经接种后,抗性变异株系H24对菌株GD3286表现抗性分离,分离比例为119∶108,其抗性遗传可能受2对互补抗病基因控制,且在SP4代仍存在抗性分离;H24SP5代株系的抗谱较原种对照显著提高,其抗谱达到84.4%,而原种对照的抗谱仅为40.6%,且H24对部分致病谱较广或专化性致病菌株表现抗病突变。经全基因组内微卫星多态性分析,H24与原种对照间未表现DNA多态性。  相似文献   

18.
Genetic diversity among 40 accessions (Coffea canephora) of robusta coffee genepool available in India was determined in comparison with 14 representative samples from a C. canephora core collection and three accessions of C. congensis, using amplified fragment length polymorphism (AFLP) and simple sequence repeats (SSR) markers. Both these molecular approaches were able to generate unique fingerprints for each of the accessions analysed. All the 12 SSR primers used in the present study were found polymorphic, with an average of six alleles per primer pair. Comparative analysis revealed the higher amount of diversity in representatives from a core collection than in the Indian genepool. Moreover, a total of 205 polymorphic AFLP bands were scored in all the 57 accessions analysed. The genetic relationship among 57 accessions was compared on the basis of SSR and AFLP polymorphisms. Genetic similarity dendrograms showed high correlation between the two marker systems. This study clearly established the high amount of diversity present in core samples, which is not represented in Indian genepool. Furthermore, the three accessions of C. congensis did not exhibit any significant diversity from other robusta accessions supporting the school of thought that C. congensis forms a biotype of C. canephora. The potential use of SSRs and AFLP markers in genetic diversity analysis for better ex situ management and also for exploitation of diversity in breeding programmes is discussed.  相似文献   

19.
A set of 24 microsatellite markers was identified in the artichoke genome, using various approaches. A genomic library allowed the development of 14 SSR markers, whereas the other 10 were obtained from gene intron/UTR regions or from other species. Allelic variation was scored in C. cardunculus (artichoke, cultivated cardoon, and wild cardoon) samples, and in other wild Cynara allies. For the 23 polymorphic loci, a total of 165 alleles were scored, 135 of which in the artichoke primary genepool, and the remaining ones in the other Cynara species. Some allele combinations were able to identify artichoke varietal types, and some alleles were unique to specific groups. This makes these markers potentially useful in product traceability and in contributing to the saturation of genetic maps. The percentage of shared alleles between C. cardunculus taxonomic groups, and Nei’s genetic distances indicated that wild cardoons from the Eastern Mediterranean were more closely related to artichoke and less to cultivated cardoon in comparison to wild cardoons from the Western Mediterranean, and the genetic distance between the two wild cardoon genepools was rather high. The UPGMA dendrogram based on Nei’s genetic distances revealed that artichokes formed a fairly defined cluster, whereas Eastern wild cardoons occupied another branch, and Western wild cardoons were clustered together with cultivated cardoons. The transferability of microsatellite markers to other Cynara wild species was quite good. Sequencing alleles at three loci showed that, apart from microsatellite length variation, point mutations and insertion/deletions were quite abundant especially when comparing C. cardunculus to the other Cynara species. In the sequenced regions, some SNPs were identified which distinguished artichoke on one side, and cultivated and wild cardoon on the other, while other SNPs were apportioned according to the geographic distribution of Cynara wild species.  相似文献   

20.
为了开发铁皮石斛EST-SSR分子标记,利用SSRIT软件对铁皮石斛近缘物种-金钗石斛的15 383条EST序列进行SSR位点查找,利用Primer 5和Oligo 7软件进行引物设计和评价,之后经PCR扩增和琼脂糖凝胶电泳对引物进行初步筛选,并通过聚丙烯酰氨凝胶电泳对所筛选的引物进行有效性检测。结果表明,SSRIT软件共查找到370条SSR位点序列,共有379个SSR位点,候选SSR位点出现的频率为2.46%。二核苷酸、三核苷酸和六核苷酸重复是最主要的重复类型,分别占41.42%、26.91%和27.44%,其中二核苷酸重复中AG/CT(20.84%)和GA/TC(18.73%)最丰富;三核苷酸重复中TCT/AGA(4.22%)和GA/TC(3.96%)最丰富;而六核苷酸重复基元较多。EST-SSR平均长度为24 bp,平均每8.5条EST序列包含1个SSR位点。设计的106对EST-SSR引物中有50对能扩增出理想的PCR产物,引物有效扩增率为47.17%,其中有43对能够扩增出多态性条带,占可扩增引物的86%。本研究开发的43对引物为铁皮石斛遗传多样性分析、种质鉴定、遗传图谱构建和功能基因研究等提供了重要的参考价值。  相似文献   

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