Intraspecific and interspecific variation in Setaria revealed by RAPD analysis

Random amplified polymorphic DNA (RAPD) analysis was applied to foxtail millet (Setaria italica) and other Setaria species in order to assess the degree of polymorphism among and within the species and to investigate their genetic relationships. Twenty accessions of foxtail millet, five of green foxtail (S. viridis), five of yellow foxtail (S. glauca), one each of S. germanica, S. verticilliformis, S. pallidifusca, S. macrostachya, S. sphacelata and S. faberi, and one unidentified, were evaluated for variability using a set of 19 random 10-mer primers. A total of 148 markers were scored for all the accessions. Wide variations in banding profiles among species were observed with nearly every primer used. The results showed that foxtail millet and S. glauca had a considerable degree of variation. The geographic differentiation of RAPD patterns in foxtail millet accessions examined seemed to exist. The present study confirmed that foxtail millet is more closely related to green foxtail than to other species in Setaria, providing support for the origin of foxtail millet from S. viridis and the conspecificity of these two species. The results confirmed that S. germanica and S. italica are conspecific and the former can be replaced by the latter. Setaria glauca and S. sphacelata are distinct from foxtail millet, implying that elite traits found in these two species are relatively difficult to be used in foxtail millet improvement. This study demonstrates that RAPD analysis can be used in assessing intraspecific and interspecific variation in foxtail millet and its wild relatives.     

Comparison of genetic variation among accessions of <Emphasis Type="Italic">Aegilops tauschii</Emphasis> using AFLP and SSR markers

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.     

不同生态区骨干谷子品种表型鉴定与遗传分析

骨干谷子品种对谷子产业发展发挥了重要的作用,为了解其优异性状遗传基础及利用价值,选用来源于不同生态区的12个骨干谷子品种,进行表型鉴定和遗传差异分析。结果表明,12个来源于不同生态区的骨干品种在株高、穗长、穗粗、穗下节间长、单穗重、生育期以及黄色素含量等方面差异较大。79对SSR多态性标记共检测到258个多态性变异位点,平均3.265 8个;SSR标记多态性(PIC)的变幅为0.141 1~0.711 6,平均为0.510 1;其中PIC>0.5的SSR标记有50对,占多态性标记的63.3%。12份材料的遗传距离变幅为0.140 2~0.801 1,平均为0.473 7;来自华北夏谷区的豫谷1号和沧谷4号遗传距离最近,而沧谷4号和来自内蒙高原的地方品种金香玉距离最远;总体而言,华北夏谷区品种和西北早熟春谷区品种遗传距离相对较大。相似性系数为0.594时,12个品种可聚为两类,华北夏谷区的豫谷1号、矮88、冀谷19号、沧谷4号和济谷12聚为一类,来自西北春谷早熟区、晚熟区和东北春谷区的7个品种聚为一类,聚类分析结果和品种的生态类型具有较高的一致性。群体结构分析和聚类分析的结果相似,华北夏谷区品种和春谷区品种之间存在基因交流。总之,不同生态区来源品种间遗传差异较大,同一生态区来源品种遗传差异较小;华北夏谷区骨干品种之间遗传差异较春谷区品种遗传差异相对较小;华北夏谷区品种和西北早熟春谷区品种间遗传差异较大,丰富华北夏谷区遗传变异,应加强与西北早熟春谷区品种之间的交流。本研究结果为优异谷子品种培育及种质资源利用提供了一定依据。     

Exploitation of <Emphasis Type="Italic">Malus</Emphasis> EST-SSRs and the utility in evaluation of genetic diversity in <Emphasis Type="Italic">Malus</Emphasis> and <Emphasis Type="Italic">Pyrus</Emphasis>

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.     

不同生态区谷子品种表型鉴定及SSR遗传多样性分析

为了明确不同生态区谷子品种的遗传多样性和遗传结构,本研究对175份来源不同的谷子品种进行29个表型性状鉴定和36对简单序列重复（SSR）标记分析。结果表明,谷子数量性状的平均遗传多样性指数最高,质量性状最低;不同类性状中,单穗码数、开花至成熟日数和粒色遗传多样性指数最高,分别为2.082 8、2.012 5和1.078 7。36对SSR引物中,共检测到401个等位基因,平均11.4个,Shannon指数和多态性信息含量（PIC）平均为2.017 2和0.810 6;标记B142、B225的Shannon指数和PIC值最高,是评价谷子遗传多样性的理想SSR标记。不同生态区表型和SSR标记多样性比较结果表明,春谷中晚熟区（ML）遗传多样性指数最高,春谷早熟区（EM）次之,夏谷区（SS）最低;春谷中晚熟区谷子品种有10个性状指标最高,且全生育期最长。春谷中晚熟区不同类型材料中,汾阳的质量性状遗传多样性指数和SSR标记Shannon指数均最高,长治数量性状遗传多样性指数最高,太原Ⅱ的生育期遗传多样性指数最高;不同类型材料间差异较低,其中7个数量性状指标差异未达显著水平;汾阳的株高最高,出谷率最...     

Genetic Variation of Blast Resistance in Foxtail Millet (<Emphasis Type="Italic">Setaria italica</Emphasis> (L.) P. Beauv.) and its Geographic Distribution

To clarify the geographic variation and isolate specificity in blast resistance, 20 cultivars of foxtail millet (Setaria italica (L.) P. Beauv.) originating from Eurasia were examined for their resistance using 11 Japanese Setaria isolates of blast fungus. Cultivars from the countries east of Pakistan generally showed resistance to most of the 11 fungus isolates, whereas those from the countries west of Afghanistan indicated higher susceptibility. The origin of this geographical regularity was discussed in relation to the deficiency or specificity of relevant resistance gene(s). No identical reaction pattern was observed among cultivars, indicating their highly distinctive isolate specificity. The virulence of isolates was also diverse, since only 2 out of 10 virulent isolates were identical for their reaction patterns. Preliminary genetic analysis of blast resistance to four fungus isolates suggests that they are governed by more than two dominant genes.     

Analysis of genetic diversity in Indian robusta coffee genepool (<Emphasis Type="Italic">Coffea canephora</Emphasis>) in comparison with a representative core collection using SSRs and AFLPs

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.     

Geographical variation of foxtail millet, Setaria italica (L.) P. Beauv. based on rDNA PCR–RFLP

The rDNA PCR–RFLP of foxtail millet (Setaria italica) germ-plasm collected throughout Eurasia and from a part of Africa was investigated with five restriction enzymes according to our previous study. Foxtail millet germ-plasms were classified by length of the rDNA IGS and RFLP; clear geographical differentiation was observed between East Asia, the Nansei Islands of Japan-Taiwan-the Philippines area, South Asia and Afghanistan-Pakistan. We also found evidence of migration of foxtail millet landraces between the areas. We calculated diversity index (D) for each region and found that center of diversity of this millet is East Asia such as China, Korea and Japan.     

rDNA polymorphism of foxtail millet (Setaria italica ssp. italica) landraces in northern Pakistan and Afghanistan and in its wild ancestor (S. italica ssp. viridis)

Ribosomal DNA (rDNA) spacer length polymorphism was studied in foxtail millet (Setaria italica ssp. italica) landraces from Pakistan and Afghanistan and in its wild ancestor (S. italica ssp. viridis) from Pakistan by PCR-based methods. Sequence polymorphism was also investigated for accessions selected based on the observed length polymorphism. The PCR-based length polymorphism and sequence polymorphism of rDNA intergenic spacer (IGS) clearly demonstrated genetic differentiation between cultivated and wild forms in the region. Genetic differentiation was observed between different areas to some extent in the cultivated form, and between different regions in the wild form of northern Pakistan. Based on the results, we discuss the genetic differentiation of foxtail millet and wild ancestor in this region and possible utility of rDNA markers to trace the dispersal of this crop in the region.     

Assessment of Genetic Diversity among Finger Millet (<Emphasis Type="Italic">Eleusine coracana</Emphasis> (L.) Gaertn.) Accessions using Molecular Markers

Finger millet (Eleusine coracana), an allotetraploid cereal, is widely cultivated in the arid and semiarid regions of the world. Being rich in protein and calcium, finger millet serves as an important staple food for rural populations in developing tropical countries where calcium deficiency and anemia are wide spread. Thirty-two finger millet genotypes were fingerprinted using 50 random amplified polymorphic DNA (RAPD) markers. Out of the total 529 loci generated using the 50 RAPD primers, 479 loci (91%) were polymorphic and informative to differentiate the accessions. Cluster analysis grouped the 32 finger millet accessions into two major clusters. Among the 32 finger millet genotypes, GEC 182 and CO 12 were distantly related with a low similarity index of 0.315. These two accessions also differed considerably in days to flowering and grain weight; GEC 182 is early flowering and has bold grains, while CO 12 is late flowering and has smaller grains. These two accessions with higher diversity at molecular level, phenology and grain weight will be ideal as parents in hybridization programme, to develop improved finger millet varieties suitable for peninsular region of India.     

Genetic diversity of the D-genome in <Emphasis Type="Italic">T. aestivum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species using SSR markers

Simple sequence repeats (SSRs), highly dispersed nucleotide sequences in genomes, were used for germplasm analysis and estimation of the genetic relationship of the D-genome among 52 accessions of T. aestivum (AABBDD), Ae. tauschii (D^tD^t), Ae. cylindrica (CCD^cD^c) and Ae. crassa (MMD^cr1D^cr1), collected from 13 different sites in Iran. A set of 21 microsatellite primers, from various locations on the seven D-genome chromosomes, revealed a high level of polymorphism. A total of 273 alleles were detected across all four species and the number of alleles per each microsatellite marker varied from 3 to 27. The highest genetic diversity occurred in Ae. tauschii followed by Ae. crassa, and the genetic distance was the smallest between Ae. tauschii and Ae. cylindrica. Data obtained in this study supports the view that genetic variability in the D-genome of hexaploid wheat is less than in Ae. tauschii. The highest number of unique alleles was observed within Ae. crassa accessions, indicating this species as a great potential source of novel genes for bread wheat improvement. Knowledge of genetic diversity in Aegilops species provides different levels of information which is important in the management of germplasm resources.     

Geographic distribution of Waxy gene SNPs and indels in foxtail millet, Setaria italica (L.) P. Beauv.

To elucidate diversity and evolution of the Waxy gene in foxtail millet, Setaria italica, we analyzed sequence polymorphism of Waxy gene in 83 foxtail millet landraces collected from various regions covering the entire geographical distribution of this millet in Europe and Asia. We found a unique geographic distribution pattern at the sequence level of gene haplotypes and also found a large diversity in East Asian landraces. We also found a higher degree of genetic polymorphism in a non-waxy phenotype than in other low amylose types, supporting the hypothesis that low amylose types recently originated from non-waxy type.     

Microsatellite markers and genetic diversity assessment in Lolium temulentum

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.     

Characterization of European hazelnut (<Emphasis Type="Italic">Corylus avellana</Emphasis>) cultivars using SSR markers

World hazelnut production is based primarily on selections from the wild. In this study, we used 21 pairs of simple sequence repeat (SSR) primers to investigate genetic diversity in 270 clonal accessions of European hazelnut (Corylus avellana) representing a wide geographic range. Of the 270 accessions, 198 had unique fingerprints while 72 were duplicates. Based on the 198 unique accessions, the number of detected alleles per locus averaged 9.81 and observed heterozygosity (H_o) averaged 0.67. Of the 206 total alleles amplified, 20 were unique to a single accession. A genetic similarity matrix was constructed and the resulting dendrogram revealed four major geographical groups: Central European, Black Sea, English and Spanish-Italian. SSR alleles indicated the parentage of 31 accessions. The fingerprints are publicly available through the Germplasm Resources Information Network (GRIN) database. The identification of duplicate and mislabeled accessions will improve management of hazelnut genebanks, and information on genetic variation in hazelnut will assist the international research community. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Geographical variation of nuclear genome RFLPs and genetic differentiation in foxtail millet, Setaria italica (L.) P. Beauv.

Sixteen RFLP loci in 62 landraces were assayed to study geneticdifferentiation in foxtail millet, Setariaitalica (L.) P. Beauv. Among 52 bands, 47 werepolymorphic among foxtail millet landraces. A dendrogram constructedbased on RFLPs was divided into five major clusters (clusterI–V). Clusters I and II contained strains mainly from EastAsia. Cluster III consisted of strains from subtropical and tropicalregions in Asia such as Nansei Islands of Japan, Taiwan and thePhilippines and India and cluster IV consisted of some strains fromEast Asia, a strain from Nepal and a strain from Myanmar. Cluster Vcontained strains from central and western regions of Eurasia such asAfghanstan, Central Asia and Europe. Chinese landraces wereclassified into four clusters. These results indicate that foxtailmillet landraces have differentiated genetically between differentregions and that Chinese landraces were highly variable.     

Simple sequence repeats marker polymorphism in emmer wheat (<Emphasis Type="Italic">Triticum dicoccon</Emphasis> Schrank): Analysis of genetic diversity and differentiation

Genetic diversity was investigated in 73 accessions of emmer wheat (Triticum dicoccon Schrank) from 11 geographical regions using a set of 29 simple-sequence repeat (SSR or microsatellite) markers, representing at least two markers for each chromosome. The SSR primers amplified a total of 357 different alleles with an average of 12.31 alleles per locus. The number of fragments detected by each primer ranged between 6 (Xgwm1066) and 21 (Xgwm268). Null alleles were detected in nine of the 29 primers used. A high level of gene diversity index was observed. Across the 29 primers, gene diversity ranged from 0.60 (Xgwm46) to 0.94 (Xgwm655), with a mean of 0.82. There was a highly significant correlation (r=0.882; p<0.01) between gene diversity index and the number of loci, showing the number of loci per se is a strong indicator of diversity. Analysis of genetic diversity within and among eleven geographical regions revealed most of the genetic diversity of the total sample resided within regions. The coefficient of gene differentiation (Gst = 0.27) showed that the genetic variation within and among the 11 geographical regions was 73 and 27%, respectively. High value of mean number of alleles per locus was found in Iran (4.86) followed by Morocco (4.10) and Armenia (4.03). On the contrary, lower mean number of alleles per locus was detected in Yemen (2.83). The average gene diversity index across regions ranged from 0.52 (Slovakia) to 0.67 (Morocco) with an average of 0.60. Multivariate techniques of principal component analysis and clustering were employed to examine genetic relationship among the 73 emmer wheat accessions vis-à-vis geographical regions of collections. The genetic distance coefficients for all possible 55 pairs of regional comparisons ranged from 0.63 (between Iran and Armenia, Georgia and Azerbaijan, Georgia and Slovakia) to 0.97 (between Morocco and Yemen, Spain and Georgia, and Turkey and Iran) with a mean of 0.82. From the PCA results, a two dimensional plot of PC1 versus PC2 was constructed. The scatter plot of the first two principal components which explained altogether 27% of the total variation depicted the presence of a clear pattern of geographical differentiation except in few cases like accessions from Caucasian region. Similar pattern of genetic relationships among accessions was observed in cluster analysis. The study provided genetic information of emmer wheat in relation to geographical regions of origin. The information could be utilized in crop improvement, germplasm conservation programs, and in further investigation.     

Assessing genetic diversity of Polish wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis>) varieties using microsatellite markers

Fifty-three wheat cultivars have been genotyped using 24 SSR (simple sequence repeat) markers in order to evaluate genetic similarities among Polish wheats, i.e. 53 spring and winter cultivars; ‘Chinese Spring’ was taken as reference. ll but one SSR marker allowed to identify DNA polymorphisms, giving in total 166 alleles (including nulls), from 3 to 13 alleles per marker with mean of 7.22. Based on marker data, genetic similarities were calculated and a dendrogram was created. ‘Spring’ cultivars were less diverse than winter ones, showing the biggest similarity to ‘Chinese Spring’. Four sister cultivars (Nutka, Tonacja, Zyta and Sukces), formed a cluster of very similar materials, of which Zyta and Sukces had the highest similarity indices. Parental lines Jubilatka and SMH 2182 were more distant from each other (genetic similarity of 0.227). It was possible to differentiate all the wheats using only four SSR markers: Xgwm186, Xgwm389, Xgwm459 and Xgwm577.     

Analysis of genetic diversity in Tunisian durum wheat cultivars and related wild species by SSR and AFLP markers

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.     

基于石蒜属转录组序列的SSR分子标记开发应用

为开发石蒜属简单重复序列(SSR)分子标记,并研究SSR引物在石蒜属内的通用性,本研究对石蒜属石蒜、忽地笑、中国石蒜、长筒石蒜、换锦花、香石蒜6个种质转录组测序,检测SSR位点并设计引物,通过PCR扩增和毛细管电泳判断引物的有效性和多态性,绘制石蒜属17个种质资源的指纹图谱并对杂交后代的真实性进行早期检测。结果表明,共获得404 481条Unigenes,利用数据库进行同源比对和功能注释,并对Unigene进行SSR位点挖掘和分析,共检测到59 612个SSR位点。其中,单核苷酸重复>二核苷酸重复>三核苷酸重复,分别占SSR总数的62.88%、20.06%和14.66%,四核苷酸及以上重复单元相对较少。选取并合成8对荧光引物进行PCR扩增,通过毛细管电泳检测发现,8对荧光引物共检测到60个多态位点,多态位点数平均为7.50,多态性信息含量(PIC)值变化范围为0.148 0~0.940 8,平均值为0.593 0。利用引物扩增带型组合法构建了石蒜属17个种资源的指纹图谱,其中引物QZ209可区分所有供试材料,并可用于杂交后代鉴定。本研究开发的SSR标记具有丰富的多态性,在石蒜属植物的资源多样性分析、杂交种鉴定及遗传图谱的构建应用中具有重要意义。