将Thinopyrum bessarabicum和Thinopyrum elongatum的种质导？…

２个双倍体即Ｃ．Ｓ－Ｔｈｉｎｏｐｙｒｕｍ ｂｅｓｓａｒａｂｉｃｕｍ（ＡＡＢＢＤＤＪＪ ２ｎ＝８ｘ＝５６）和ＧＨＫ－Ｔｈｉｎｏｐｙｒｕｍ ｅｌｏｎｇａｔｕｍ（ＡＡＢＢＤＤＥＥ ２ｎ＝８ｘ＝５６）与普通小麦“中国春”杂交,获得了２个七倍体杂种。对２３个双倍体、中国春、杂种Ｆ１和部分Ｆ２进行麦谷蛋白ＳＤＳ－Ｐａｇｅ电泳,及结合染色体分带技术和田间赤霉病抗性鉴定筛选出由Ｔｈ．ｂｅｓｓａｒａｂｉｃｕｍ和Ｔｈ     

Synthesis and cytological characterization of trigeneric hybrids involving durum wheat,Thinopyrum bessarabicum,and Lophopyrum elongatum

Summary In an attempt to transfer genes for salt tolerance and other desirable traits from the diploid wheatgrasses, Thinopyrum bessarabicum (2n=2x=14; JJ genome) and Lophopyrum elongatum (2n=2x=14; EE genome), into durum wheat cv Langdon (2n=4x=28; AABB genomes), trigeneric hybrids with the genomic constitution ABJE were synthesized and cytologically characterized. C-banding analysis of somatic chromosomes of the A, B, J, and E genomes in the same cellular environment revealed distinct banding patterns; each of the 28 chromosomes could be identified. They differed in the total amount of constitutive heterochromatin. Total surface area and C-banded area of each chromosome were calculated. The B genome was the largest in size, followed by the J, A, and E genomes, and its chromosomes were also the most heavily banded. Only 25.8% of the total chromosome complement in 10 ABJE hybrids showed association, with mean arm-pairing frequency (c) values from 0.123 to 0.180 and chiasma frequencies from 3.36 to 5.02 per cell. The overall mean pairing was 0.004 ring IV + 0.046 chain IV + 0.236 III + 0.21 ring II + 2.95 rod II + 20.771. This is total pairing between chromosomes of different genomes, possibly between A and B, A and J, A and E, B and J, B and E, and J and E, in the presence of apparently functional pairing regulator Ph1. Because chromosome pairing in the presence of Ph1 seldom occurs between A and B, or between J and E, it was inferred that pairing between the wheat chromosomes and alien chromosomes occurred. The trigeneric hybrids with two genomes of wheat and one each of Thinopyrum and Lophopyrum should be useful in the production of cytogenetic stocks to facilitate the transfer of alien genes into wheat.     

Multidisciplinary approach to genome analysis in the diploid species,Thinopyrum bessarabicum and Th. elongatum (Lophopyrum elongatum), of the Triticeae

Summary The J and E genome species of the Triticeae are invaluable sources of salt tolerance. The evidence concerning the phyletic relatedness of the J genome of diploid Thinopyrum bessarabicum and the E genome of diploid Th. elongatum (=Lophopyrum elongatum) is discussed. Low level of chromosome pairing between J and E at different ploidy levels, suppression of J-E pairing by the Ph1 pairing regulator that inhibits homoeologous pairing, complete sterility of the diploid hybrids (JE), karyotypic divergence of the two genomes, differences in total content and distribution of heterochromatin along their chromosomes, and marked differences in gliadin proteins, isozymes, 5S DNA, and rDNA indicate that J and E are distinct genomes. Well-defined biochemical markers have been identified in the two genomes and may be useful in plant breeding. The level of distinction between J and E is comparable to that among the universally accepted homoeologous genomes A, B, and D of wheat. Therefore, the J and E genomes are homoeologous and not homologous, although some workers continue to call them homologous. The previous workers' data on chromosome pairing in diploid hybrids and/ or karyotypic differences in the conventionally stained chromosomes do not provide sufficient evidence for the proposed merger of J and E genomes (and, hence, of the genera Thinopyrum and Lophopyrum) specifically and for establishing genome relationships generally. Extra precautions should be exercised before changing the designation of an established genome and before merging two genera. A uniform, standardized system of genomic nomenclature for the entire Triticeae is proposed, which should benefit cytogeneticists, plant breeders, taxonomists, and evolutionists.Cooperative investigations of the USDA-Agricultural Research Service and the Utah Agricultural Experiment Station, Logan, UT 84322, USA. Approved as Journal Paper no. 3832     

Genetic diversity in wild Spanish populations of Thinopyrum junceum and Thinopyrum junceiforme using endosperm proteins and PCR-based markers

The genetic variation of sixteen wild, Spanish populations of Thinopyrum junceum and Thinopyrumjunceiforme and their interspecific relationships were analyzed. The relationships between these species and the diploids T. bessarabicum and T. elongatum were also investigated. The number of phenotypes and the composition of bands yielded by the electrophoretic separation of endosperm proteins were used to estimate intra- and interpopulational variability. DNA polymorphism generated by 24 arbitrary 10-mer primers and 14 specific 20-mer primers was used to determine interpopulational variability and interspecific relationships. Jaccard's coefficient of similarity was used to analyze presence and absence data in the DNA polymorphism and endosperm protein determinations of individual plants. Pearson's product-moment correlation coefficient was used to analyse interpopulational variation using endosperm protein band frequency data. Dendrograms were constructed using an unweighted pair group method with arithmetical average (UPGMA). The high level of intrapopulational variability found in T. junceum and T. junceiforme was inconsistent with the traditional classification of these species as self-pollinating. The level of interpopulational variation varied according to the degree of polymorphism of the corresponding markers. The endosperm proteins and random amplified polymorphic DNAs (RAPDs) proved to be the most polymorphic markers to those used although only the former were able to distinguish between the different populations. Interspecific relationships were consistently confirmed by all the PCR-based markers, and were also in agreement with the results of other authors.     

Biochemical and molecular diagnostics of Thinopyrum bessarabicum chromosomes in Triticum aestivum germ plasm

Thinopyrum bessarabicum (2n=2x=14, JJ) is a self-fertile salt-tolerant grass species, and its hybridization with Triticum aestivum to achieve the transfer of this attributes has been promoted. For the detection of alien introgression, development of diagnostic markers of Th. bessarabicum chromosomes in the wheat background has emerged as an important aspect in our intergeneric hybridization program. Six proteins/isozymes-high-molecular-weight glutenins, superoxide dismutase, grain esterase, β-amylase, glutamate oxaloacetate transaminase and α-amylase —were identified as positive markers for detecting the presence of Th. bessarabicum chromosomes in the advanced backcross derivatives of T. aestivum/Th. bessarabicum//n^* T. aestivum. Fluorescent in situ hybridization further enabled the detection of complete and translocated arms of Th. bessarabicum chromosomes in the T. aestivum background. These diagnostic markers served for tentatively characterizing a distinct set of Th. bessarabicum disomic additions to wheat (2n=44) and have facilitated establishing the homoeology of these added chromosomes.     

Genome constitutions of Thinopyrum curvifolium, T. scirpeum, T. distichum, and T. junceum (Triticeae: Gramineae).

The objective of this study is to elucidate genome constitutions of Thinopyrum curvifolium (Lange) D.R. Dewey, T. scirpeum (K. Presl) D.R. Dewey, T. distichum (Thunb.) A. L?ve, and T. junceum (L.) A. L?ve. Hybrids of T. sartorii (Boiss. &Heidr.) A. L?ve with T. scirpeum and T. junceum, as well as the hybrid between T. curvifolium and Pseudoroegneria geniculata ssp. scythica (Nevski) A. L?ve, were made and chromosome pairing at metaphase I was studied. The karyotype analyses of mitotic cells stained by aceto-orcein were conducted for both hybrids and the four target species. The Giemsa C-banding following acetocarmine staining was carried out for the above species and the triploid hybrid T. curvifolium x T. bessarabicum (Savul &Rayss) A. L?ve. Meiotic data indicate that all target species have two sets of the basic genome J, but they behave like true allopolyploids because of bivalentization. Karyotypes of T. curvifolium and its triploid hybrid with T. bessarabicum indicate that T. curvifolium contains two different versions of the Jb genome, designated as Jb3 and Jb4, rather than two Je genomes as previously believed. Thinopyrum scirpeum and T. elongatum (4x) have similar karyotypes. Both are segmental allotetraploids carrying two forms of the Je genome. Their genome formulae are Je2 Je3 and Je1 Je3, respectively. Thinopyrum distichum has a karyotype similar to T. junceiforme, which has the Jb2 Je2 genome formula. However, the two species differ in C-banding patterns, reflecting their geographical separation. Thinopyrum junceum is a hexaploid with two pairs of Jb2 genomes and one pair of the Je2 genome, and it has a C-banding pattern similar to that of T. junceiforme, which has one pair each of the Jb2 and Je2 genomes.     

四倍体小麦背景中长穗偃麦草E染色体传递特征

通过细胞学方法和染色体特异分子标记鉴定六倍体小偃麦(AABBEE)与硬粒小麦(AABB)杂交的自交后代F₂和F₃植株,探讨长穗偃麦草染色体在硬粒小麦背景中世代间的传递特征,并筛选硬粒小麦-长穗偃麦草E染色体附加系。对218个F₂单株染色体数检测表明,2n=28植株占41.7%,2n=29植株占18.3%,其余40.0%植株的染色体数在2n=31~42范围内。分子标记鉴定表明,在F₂代2n=29单体附加植株中,不同的长穗偃麦草染色体传递率之间存在明显差异,1E传递率最高,3E和6E传递率最低。在F₂代2n=30单株中,1E、4E、7E和5E染色体相互组合产生的双单体多,6E参与组合较少,未检测到2E或3E与其他染色体的组合单株。在1E~7E单体附加株自交后代F₃中,E染色体传递率变化范围为9.1%~27.5%,1E传递率最高,6E传递率最低,与F₂的传递率一致。从F₃代中选育出1E~7E单体附加及少数二体附加,所有单体附加均可育。这些附加E染色体材料将对小麦代换系和易位系的创制提供有益的中间材料。     

Genomic rearrangement between wheat and Thinopyrum elongatum revealed by mapped functional molecular markers

Thinopyrum elongatum serves as an excellent gene pool for wheat improvement. Genes for resistance to many biotic and abiotic stresses have been transferred from Th. elongatum to wheat through chromosome manipulation. For breeding programs, molecular markers enable screening of a large number of genotypes for alien chromosome introgressions. The main objective of the present study was to develop and characterize EST (expressed sequence tags) and PLUG (PCR-based Landmark Unique Gene) markers that can distinguish Th. elongatum chromatin from the wheat genomes. A total of 258 mapped EST primer pairs and 46 PLUG primer pairs were tested on DNA from wheat Chinese Spring (CS) and CS-Th. elongatum addition lines. The results showed that 43 primer pairs could be effectively mapped to specific Th. elongatum chromosomes. Twenty-two of the 43 markers displayed similar homoeologous chromosome locations to hexaploid wheat. Nine markers mapped to different linkage groups between wheat and Th. elongatum, while 12 makers mapped on two or three different Th. elongatum chromosomes. A comparison of molecular marker locations indicated that Th. elongatum genome was closely related to the D genome of wheat, and chromosome rearrangements and duplication had occurred in Th. elongatum and the wheat genomes. The markers will be useful in comparative gene mapping, chromosome evolutionary analysis, and gene introgression for wheat improvement using Th. elongatum accessions as gene donors.     

基于高通量测序的长穗偃麦草功能分子标记发掘和分析

长穗偃麦草是小麦重要的近源物种,含有丰富的抗逆基因,广泛地应用于小麦的遗传改良育种。本研究利用高通量测序,获得长穗偃麦草的转录组测序信息,利用比较基因组学方法研究其与小麦、水稻和玉米等作物的遗传关系,评估它们之间的亲缘关系。同时,将长穗偃麦草的高通量序列比对到小麦基因,利用软件Freebayes和SAMtools/Bcftools发掘功能基因的变异位点,并对这些含有变异位点的功能基因进行注释分析,揭示长穗偃麦草优异性状形成的分子机制,这将为长穗偃麦草优异基因资源的开发和应用奠定重要基础。     

Thinopyrum bessarabicum: biochemical and cytological markers for the detection of genetic introgression in its hybrid derivatives with Triticum aestivum L.

Summary Thinopyrum bessarabicum (2n = 2x = 14, JJ) with its unique property of salt tolerance provides a potential means for the transfer of this important and complex trait into cultivated wheat through intergeneric hybridization. To accomplish this, diagnostic markers for detecting the presence of Th. bessarabicum chromosomes in a wheat background have to be established. The C-banded karyotype of Th. bessarabicum distinctly identifies individual Th. bessarabicum chromosomes and separates them from those of Triticum aestivum. Also, seven protein/isozymes, i.e., malate dehydrogenase, high-molecular-weight glutenin, Superoxide dismutase, grain esterase, glutamate oxaloacetate transaminase, -amylase and -amylase, were identified as being positive markers specific to Th. bessarabicum; these were also expressed in the T. aestivum/Th. bessarabicum amphiploid. These diagnostic biochemical markers could be useful in detecting and establishing homoeology of Th. bessarabicum chromosomes in T. aestivum/Th. bessarabicum intergeneric hybrid derivatives.     

长穗偃麦草优异基因的染色体定位及应用

长穗偃麦草比较公认的有2个种,即二倍体长穗偃麦草(Thinopyrum elongatum,2n=2X)和十倍体长穗偃麦草(Thinopyrum ponticum,2n=10X),是重要的小麦近缘种,具有抗病、抗寒、抗旱、耐盐碱等优良性状。因其基因组中蕴含许多对小麦品种改良极为有用的基因,且易与小麦杂交等优势,多年来长穗偃麦草一直作为小麦遗传改良的优良种质资源而备受关注。本文对长穗偃麦草的基因组研究及其在小麦的抗逆、抗病和提高光合能力、产量及高分子量谷蛋白(HMW-GS)含量等方面的应用做了综述,为其基因组中优异基因的进一步开发和利用提供了理论依据。     

Introduction of Thinopyrum intermedium ssp. trichophorum chromosomes to wheat by trigeneric hybridization involving Triticum,Secale and Thinopyrum genera

Main conclusion

Fluorescence in situ hybridization and molecular markers have confirmed that several chromosomes from Thinopyrum intermedium ssp. trichophorum have been added to a wheat background, which originated from a cross between a wheat– Thinopyrum partial amphiploid and triticale. The lines displayed blue grains and resistance to wheat stripe rust.

Thinopyrum intermedium has been used as a valuable resource for improving the disease resistance and yield potential of wheat. With the aim to transfer novel genetic variation from Th. intermedium species for sustainable wheat breeding, a new trigeneric hybrid was produced by crossing an octoploid wheat–Th. intermedium ssp. trichophorum partial amphiploid with hexaploid triticale. Fluorescence in situ hybridization (FISH) revealed that Thinopyrum chromosomes were transmitted preferably and the number of rye chromosomes tended to decrease gradually in the selfed derivatives of the trigeneric hybrids. Four stable wheat–Th. intermedium chromosome substitution, addition and translocation lines were selected, and a 2J^S addition line, two substitution lines of 4J^S(4B) and 4J(4B), and a small 4J.4B translocation line were identified by FISH and molecular markers. It was revealed that the gene(s) responsible for blue grains may located on the FL0.60–1.00 of long arm of Th. intermedium-derived 4J chromosome. Disease resistance screenings indicated that chromosomes 4J^S and 2J^S appear to enhance the resistance to stripe rust in the adult plant stage. The new germplasm with Th. intermedium introgression shows promise for utilization of Thinopyrum chromosome segments in future wheat improvement.

     

RAPD and ISSR-assisted identification and development of three new SCAR markers specific for the Thinopyrum elongatum E (Poaceae) genome

Diploid Thinopyrum elongatum, a wild relative of wheat, contains many agronomically desirable traits and has potential for increasing genetic variability and introducing desirable characters in this crop. Few molecular markers are available for rapid screening of T. elongatum genome segments in the wheat genetic background. We used 36 RAPD primers and 33 ISSR primers to screen for polymorphisms in the common wheat variety Chinese Spring and in T. elongatum. Two RAPD markers and one ISSR marker, designated OPF03(1407), LW10(1487) and UBC841(701), were identified and were specific for the T. elongatum E genome. Three pairs of primers flanking these specific sequences were designed to produce SCAR markers. All three SCAR markers were T. elongatum E genome-specific. Two of these SCAR markers, SCAR(807) and SCAR(577), were present in all seven T. elongatum chromosomes, while SCAR(839) was specific for T. elongatum chromosomes 2E and 3E. These newly developed SCAR markers should be useful for detecting alien genome chromatin or chromosome segments in the genetic background of common wheat.     

Molecular Cytogenetics of an Amphiploid Trigeneric Hybrid between Triticum durum, Thinopyrum distichum and Lophopyrum elongatum

In situ hybridization of total genomic DNA was used to analyselines derived from an amphiploid between tetraploid wheat,Triticumdurum Desf. (2n =4x =28), and the wheatgrassesThinopyrum distichum(Thunb.) A. Löve (2n =4x =28) andLophopyrum elongatum (Host)A. Löve (2n =2x =14). A range of chromosome numbers wasdetected, arising from loss or gain of chromosomes. Total genomicDNA probes fromThinopyrum species,L. elongatum andTriticum monococcumL. were able to discriminate chromosomes from the A and B genomesof tetraploid wheat and those of wheatgrass-origin. The methoddid not discriminate the two wheatgrass genomes, J and E, indicatingtheir close similarity. Chromosomal aberrations—includingtelocentric and ring chromosomes—were frequent. Distalinter-genomic translocations of parts of A and B genome chromosomearms, unusual in wheat itself, were more frequent than translocationsbetweenT. durum and wheatgrass.In situ hybridization of an rDNAprobe most frequently revealed four sites associated with secondaryconstrictions onT. durum chromosomes and four onTh. distichumorL. elongatum chromosomes, although there was variation inthe number of loci between and within plants. Within interphaseand prophase nuclei, the three genomes were not intermixed andoften lay in distinct sectors. Wheat; hybrids; Triticum ; Triticeae; evolution; introgression; nuclear architecture; rDNA; in situ hybridization     

Salt Tolerance in the Triticeae: Growth and Solute Accumulation in Leaves of Thinopyrum bessarabicum

Gorham, J., McDonnell, E., Budrewicz, E. and Wyn Jones, R. G.1985. Salt tolerance in the Triticeae: growth and solute accumulationin leaves of Thinopyrum bessarabicum.—J. exp. Bot. 36:1021–1031. The diploid wheatgrass Thinopyrum bessarabicum was found towithstand prolonged exposure to 350 mol m^–3 NaCl in hydroponicculture. During the gradual addition of salt to the externalmedium, osmotic adjustment was rapidly achieved by the accumulationof Na and Cl. Following osmotic adjustment constant leaf Naand Cl concentrations were maintained, and K was retained ata high level. Thinopyrum bessarabicum may be described as anosmoconformer, adjusting its internal osmotic pressure to 400–500mOsmol kg^–1 above that of the external medium in hydroponicculture. Both slower shoot initiation and reduced leaf lengthcontributed to the reduced growth rates at higher salinities.Leaf width was not affected. Increasing salinity resulted inincreases in leaf concentrations of phosphate, glycinebetaine,sucrose and proline, and in decreases in the concentrationsof nitrate, sulphate, magnesium, calcium, total amino acidsand organic acids. Thinopyrum bessarabicum exhibits salt tolerancecharacters which may be useful in wheat breeding. Key words: Salt stress, solute accumulation, osmotic adjustment, Thinopyrum     

Characterization of Thinopyrum bessarabicum chromosome segments in wheat using random amplified polymorphic DNAs (RAPDs) and genomic in situ hybridization

Ten random amplified polymorphic DNA (RAPD) markers specific to chromosome 5E^b of Thinopyrum bessarabicum were detected. Genomic in situ hybridization and standard cytological observations revealed that six of the markers are located on the 5E^b short arm and four are located on the 5E^b long arm. These RAPD markers have been used to confirm the identity of putative 5E^b (5A) and 5E^b (5D) substitution individuals. The potential of RAPDs for the detection of wheat/alien recombinants is discussed.     

Analysis of hybrids of durum wheat with Thinopyrum junceiforme using RAPD markers

 The objective of this study was to detect the presence of alien chromatin in intergeneric hybrids of durum wheat (Triticum turgidum, 2n=4x=28; AABB genomes) with the perennial grass Thinopyrum junceiforme (2n=4x=28; J₁J₁J₂J₂) using RAPD markers. The first step was to identify amplification of species-specific DNA markers in the parental grass species and durum wheat cultivars. Initially, the genomic DNA of five grass species (Thinopyrum junceiforme, Th. bessarabicum, Lophopyrum elongatum, Leymus karataviensis and Elytrigia pycnantha) and selected durum cultivars (‘Langdon’, ‘Durox’, ‘Lloyd’, ‘Monroe’, and ‘Medora’) was screened with 40 oligonucleotide primers (nano-mers). Three oligonucleotides that amplified DNA fragments specific to a grass species or to a durum cultivar were identified. Primer PR21 amplified DNA fragments specific to each of the five durum cultivars, and primers PR22 and PR23 amplified fragments specific to each of the grass species. Intergeneric hybrids between the durum cultivars ‘Langdon’, ‘Lloyd’ and ‘Durox’ and Th. junceiforme, and their backcross (BC) progeny were screened with all 40 primers. Six primers amplified parent-specific DNA fragments in the F₁ hybrids and their BC₁ progeny. Three primers, PR22, PR23 and PR41, that amplified Th. junceiforme DNA fragments in both F₁ and BC₁ were further analyzed. The presence of an amplified 1.7-kb Th. junceiforme DNA fragment in the F₁ hybrids and BC₁ progeny was confirmed using Southern analysis by hybridization with both Th. junceiforme genomic DNA and Th. junceiforme DNA amplified with primer PR41. With the exception of line BC₁F₂ no. 5, five selfed progeny of BC₁ and a BC₂ of line 3 (BC₁F₂ no. 3בLloyd’) from a cross of ‘Lloyd’×Th. junceiforme showed the presence of the 1.7-kb DNA fragment. All selfed BC₁ and BC₂ lines retained the 600-bp fragment that was confirmed after hybridization with Th. junceiforme DNA amplified with primer PR22. Other experiments using RFLP markers also showed the presence of up to seven Th. junceiforme DNA fragments in the F₁ hybrids and their BC progeny after hybridization with Th. junceiforme DNA amplified with primer PR41. These studies show the usefulness of molecular markers in detecting alien chromatin/DNA fragments in intergeneric hybrids with durum wheat. Received: 21 November 1996 / Accepted: 21 March 1997     

Analysis of diversity and relationships among Chinese orchid cultivars using EST-SSR markers

Chinese orchid (Cymbidium spp.) is an important potted flower with extremely high ornamental value in China. It is important to identify the genetic diversity of its germ plasma resources for development and evaluation of its cultivars. A set of 13 expressed sequence tag (EST)-derived simple sequence repeat (SSR) was used to analyze 103 cultivars of six species of Chinese orchid, namely Cymbidium goeringii, C. faberi, Cymbidium ensifolium, C. kanran, C. sinense and C. goeringii var. longibracteatum. The 13 SSR primer pairs generated a total of 168 polymorphic bands, with an average of 12.92 bands per primer and a range of 6–24 bands which clearly revealed the difference between cultivars inter- or intra-species of Chinese orchid. Cluster analysis based on UPGMA, NJ and PCoA method showed a dendrogram with three basic clusters and splitting feature of C. ensifolium and C. goeringii which partially congruent with the current taxonomic classification.     

Assessment of genetic diversity in the sorghum reference set using EST-SSR markers

Selection and use of genetically diverse genotypes are key factors in any crop breeding program to develop cultivars with a broad genetic base. Molecular markers play a major role in selecting diverse genotypes. In the present study, a reference set representing a wide range of sorghum genetic diversity was screened with 40 EST-SSR markers to validate both the use of these markers for genetic structure analyses and the population structure of this set. Grouping of accessions is identical in distance-based and model-based clustering methods. Genotypes were grouped primarily based on race within the geographic origins. Accessions derived from the African continent contributed 88.6 % of alleles confirming the African origin of sorghum. In total, 360 alleles were detected in the reference set with an average of 9 alleles per marker. The average PIC value was 0.5230 with a range of 0.1379–0.9483. Sub-race, guinea margaritiferum (Gma) from West Africa formed a separate cluster in close proximity to wild accessions suggesting that the Gma group represents an independent domestication event. Guineas from India and Western Africa formed two distinct clusters. Accessions belongs to the kafir race formed the most homogeneous group as observed in earlier studies. This analysis suggests that the EST-SSR markers used in the present study have greater discriminating power than the genomic SSRs. Genetic variance within the subpopulations was very high (71.7 %) suggesting that the germplasm lines included in the set are more diverse. Thus, this reference set representing the global germplasm is an ideal material for the breeding community, serving as a community resource for trait-specific allele mining as well as genome-wide association mapping.