Allelic reduction and genetic shift in the Canadian hard red spring wheat germplasm released from 1845 to 2004

Analysis of genetic diversity changes in existing gene pools of cultivated crops is important for understanding the impact of plant breeding on crop genetic diversity and developing effective indicators for genetic diversity of cultivated plants. The objective of this study was to assess genetic diversity changes in 75 Canadian hard red wheat (Triticum aestivum L.) cultivars released from 1845 to 2004 using 31 simple sequence repeats (SSRs) markers. A total of 267 SSR alleles were detected, and their allelic frequencies ranged from 0.01 to 0.97, with an average of 0.14. Significant allelic reduction was observed at only four SSR loci for the cultivars released from 1970 onwards. However, 51 alleles (about 19%) present in pre-1910 cultivars were undetected in cultivars released after 1990 and were spread over 27 SSR loci. The proportion of SSR variation accounted for by six breeding periods was 12.5%, by four ancestral families, 16.5%, and by eight breeding programs, 8.4%. The average genetic diversity measured by three different band-sharing methods did not change significantly among cultivars released from different breeding periods, breeding programs, and ancestral families. However, genetic shift was obvious in the cultivars released over the six breeding periods, reflecting well the various breeding efforts over years. These results clearly show the allelic reduction and genetic shift in the Canadian hard red spring wheat germplasm released over time. Consequently, more effort needs to be made to broaden the wheat breeding base and conserve wheat germplasm.     

部分耐盐小麦品种(系)SSR位点遗传多样性研究

选择有多态性的32对SSR引物对80个小麦耐盐品种(系)进行遗传差异研究,共检测出155个等位变异,平均每个位点上有4.75个等位变异;供试80份耐盐小麦品种(系)来源广泛,遗传基础丰富,表现出较高的遗传多样性,遗传相似系数范围在0.26～0.81;聚类分析结果显示,冬性小麦品种(系)聚为一大类;春性小麦品种(系)也聚为一大类;一些系谱相同或相近的品种(系)遗传相似系数较大;A、B、D基因组中SSR位点平均等位变异差异不大,以B基因组较高.     

EST-SSR DNA polymorphism in durum wheat (Triticum durum L.) collections

SSRs derived from EST were molecular markers belonging to the transcribed region of the genome. Therefore, any polymorphism detected using EST-SSRs might reflect the better relationship among species or varieties. Using wheat EST-SSR markers, 60 durum wheat (Triticum durum L.) accessions from seven countries were investigated. Twenty-five primer pairs could amplify successfully in the 60 durum wheat accessions, of which tri-nucleotide repeats were the dominant type, and revealed 26 loci on all seven wheat homologous chromosome groups. A total of 87 eSSR alleles were detected, and the number of alleles detected by a single pair of primers ranged from 1 to 11, with an average of 3.3 alleles per locus. Higher numbers of alleles and PIC were identified on the B genome than those on the A genome.     

Diversity changes in an intensively bred wheat germplasm during the 20th century

Plant breeding may lead to narrowing genetic diversity of cultivatedcrops, thereby affecting sustained selection gains in crop improvement. A totalof 47 microsatellite primer pairs (mapped to the 21 wheat genetic linkagegroups) were assessed in 75 Nordic spring wheat cultivars bred during the20^th century to determine the variation of genetic diversity in thisgermplasm throughout this period. The number of alleles ranged from one toseven, with an average of 3.6 alleles per microsatellite marker. A dendrogramresulting from analysis of the matrix of dissimilarities using the unweightedpair-group method with arithmetic average discriminated all cultivars andrevealed clusters of accessions released both from some geographical area inthe Nordic Region and the breeding era, i.e. before and after World War II. Geneticdiversity in this wheat material increased from 1900 to 1940 and again from1960 onwards. In between these two periods there was a loss of diversity, whichcould not be explained by changes in a single genome or in one or few chromosomesets. Effects of different selection within countries are revealed by cleardifferences in frequency of some microsatellite alleles. In adition somemicrosatellite alleles were lost during the first quarter of the century whileseveral new alleles were introduced in the Nordic spring wheat material duringthe second half of the century. These results suggest that genetic diversity inNordic spring wheat was enhanced by plant breeding in the first quarter of the20^th century and following a decrease during the second quarter wasincreased again by plant breeding.     

Comparative analysis of the genetic diversity dynamics at gliadin loci in the winter common wheat Triticum aestivum L. cultivars developed in Serbia and Italy over 40 years of scientific breeding

The dynamics of genetic transformations at gliadin-coding loci in the winter common wheat cultivars produced in Serbia and Italy over 40 years of scientific breeding was studied. It was demonstrated that a number of alleles unique for the wheat cultivars of each country were substituted with the alleles of a limited number of donor cultivars, in particular, cultivar Bezostaya 1 and its derivatives. On the background of preserved heterogeneity values during various time periods of breeding in each country, the genetic diversity in the total region decreased, as demonstrated by similarity in the sets of alleles of gliadin loci and their frequencies in the modern cultivars of these two Southern European countries. This decrease in the genetic diversity is an erosion of genetic resources within the region, which results in a loss of unique coadapted gene complexes.     

Comparative analysis of the genetic diversity dynamics at gliadin loci in the winter common wheat Triticum aestivum L. cultivars developed in Serbia and Italy over 40 years of scientific breeding

The dynamics of genetic transformations at gliadin-coding loci in the winter common wheat cultivars produced in Serbia and Italy over 40 years of scientific breeding was studied. It was demonstrated that a number of alleles unique for the wheat cultivars of each country were substituted with the alleles of a limited number of donor cultivars, in particular, cultivar Bezostaya 1 and its derivatives. On the background of preserved heterogeneity values during various time periods of breeding in each country, the genetic diversity in the total region decreased, as demonstrated by similarity in the sets of alleles of gliadin loci and their frequencies in the modem cultivars of these two Southern European countries. This decrease in the genetic diversity is an erosion of genetic resources within the region, which results in a loss of unique coadapted gene complexes.     

用EST-SSR检测不同年代小麦育成品种基因多样性的变化趋势

EST—SSRs是一种基于基因表达序列有关基因功能的“真质”标记,其多态性直接反映了基因的多样性。本研究采用37对小麦EST—SSRs引物检测了42份我国不同年代小麦选育品种相关基因位点多样性的变化趋势。结果表明：基于表达序列设计的小麦EST—SSRs引物具有较好的扩增效果,其中37对引物共检测到134个位点,绝大多数引物有2～5个等位变异;在相似系数为0.9的水平上这些引物可将除燕大1817和旱选3号外的40份材料区分开来;供试的42份小麦选育品种的基因多样性从20世纪60年代以前到90年代呈递增趋势,但不同年代的增幅不同：20世纪60～80年代增长最快,80年代至90年代增长十分缓慢;同时,材料间相似系数变化呈下降趋势,60年代以前的选育品种相似性最高,70年代最低,70年代以后至90年代略有增加,但远低于60年代以前,这与引人大量的外来品种的育种策略有关。     

Wheat genetic diversity trends during domestication and breeding

It has been claimed that plant breeding reduces genetic diversity in elite germplasm which could seriously jeopardize the continued ability to improve crops. The main objective of this study was to examine the loss of genetic diversity in spring bread wheat during (1) its domestication, (2) the change from traditional landrace cultivars (LCs) to modern breeding varieties, and (3) 50 years of international breeding. We studied 253 CIMMYT or CIMMYT-related modern wheat cultivars, LCs, and Triticum tauschii accessions, the D-genome donor of wheat, with 90 simple sequence repeat (SSR) markers dispersed across the wheat genome. A loss of genetic diversity was observed from T. tauschii to the LCs, and from the LCs to the elite breeding germplasm. Wheats genetic diversity was narrowed from 1950 to 1989, but was enhanced from 1990 to 1997. Our results indicate that breeders averted the narrowing of the wheat germplasm base and subsequently increased the genetic diversity through the introgression of novel materials. The LCs and T. tauschii contain numerous unique alleles that were absent in modern spring bread wheat cultivars. Consequently, both the LCs and T. tauschii represent useful sources for broadening the genetic base of elite wheat breeding germplasm.     

Analysis of gene-derived SNP marker polymorphism in US wheat (Triticum aestivum L.) cultivars

In this study, we developed 359 detection primers for single nucleotide polymorphisms (SNPs) previously discovered within intron sequences of wheat genes and used them to evaluate SNP polymorphism in common wheat (Triticum aestivum L.). These SNPs showed an average polymorphism information content (PIC) of 0.18 among 20 US elite wheat cultivars, representing seven market classes. This value increased to 0.23 when SNPs were pre-selected for polymorphisms among a diverse set of 13 hexaploid wheat accessions (excluding synthetic wheats) used in the wheat SNP discovery project (). PIC values for SNP markers in the D genome were approximately half of those for the A and B genomes. D genome SNPs also showed a larger PIC reduction relative to the other genomes (P < 0.05) when US cultivars were compared with the more diverse set of 13 wheat accessions. Within those accessions, D genome SNPs show a higher proportion of alleles with low minor allele frequencies (<0.125) than found in the other two genomes. These data suggest that the reduction of PIC values in the D genome was caused by differential loss of low frequency alleles during the population size bottleneck that accompanied the development of modern commercial cultivars. Additional SNP discovery efforts targeted to the D genome in elite wheat germplasm will likely be required to offset the lower diversity of this genome. With increasing SNP discovery projects and the development of high-throughput SNP assay technologies, it is anticipated that SNP markers will play an increasingly important role in wheat genetics and breeding applications. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Genetic structure of wild emmer wheat populations as reflected by transcribed versus anonymous SSR markers.

Simple sequence repeat (SSR) markers have become a major tool in population genetic analyses. The anonymous genomic SSRs (gSSRs) have been recently supplemented with expressed sequence tag (EST) derived SSRs (eSSRs), which represent the transcribed regions of the genome. In the present study, we used 8 populations of wild emmer wheat (Triticum turgidum subsp. dicoccoides) to compare the usefulness of the two types of SSR markers in assessing allelic diversity and population structure. gSSRs revealed significantly higher diversity than eSSRs in terms of average number of alleles (14.92 vs. 7.4, respectively), polymorphic information content (0.87 vs. 0.68, respectively), and gene diversity (He; 0.55 vs. 0.38, respectively). Despite the overall differences in the level of diversity, Mantel tests for correlations between eSSR and gSSR pairwise genetic distances were found to be significant for each population as well as for all accessions jointly (RM=0.54, p=0.01). Various genetic structure analyses (AMOVA, PCoA, STRUCTURE, unrooted UPGMA tree) revealed a better capacity of eSSRs to distinguish between populations, while gSSRs showed a higher proportion of intrapopulation (among accessions) diversity. We conclude that eSSR and gSSR markers should be employed in conjunction to obtain a high inter- and intra-specific (or inter- and intra-varietal) distinctness.     

Allele-specific amplification of polymorphic sites for the detection of powdery mildew resistance loci in cereals

Primers for the polymerase chain reaction (PCR) were tailored to selectively amplify RFLP marker alleles associated with resistance and susceptibility for powdery mildew in cereals. The differentiation between marker alleles for susceptible and resistant genotypes is based on the discrimination of a single nucleotide by using allele-specific oligonucleotides as PCR primers. The PCR assays developed are diagnostic for RFLP alleles at the loci MWG097 in the barley genome and Whs350 in the wheat genome. The first marker locus is closely linked to MlLa resistance in barley, while the latter is linked to Pm2 resistance locus in wheat. PCR analysis of 31 barley and 30 wheat cultivars, with some exceptions, verified the presence or absence of the resistance loci investigated. These rapid PCR-based approaches are proposed as an efficient alternative to conventional procedures for selecting powdery mildew-resistant genotypes in breeding programs.     

Molecular characterization of vernalization response genes in Canadian spring wheat.

Vernalization response (Vrn) genes play a major role in determining the flowering/maturity times of spring-sown wheat. We characterized a representative set of 40 western Canadian adapted spring wheat cultivars/lines for 3 Vrn loci. The 40 genotypes were screened, along with 4 genotypes of known Vrn genes, using previously published genome-specific polymerase chain reaction primers designed for detecting the presence or absence of dominant or recessive alleles of the major Vrn loci: Vrn-A1, Vrn-B1, and Vrn-D1. The dominant promoter duplication allele Vrn-A1a was present in 34 of 40 cultivars/lines, whereas the promoter deletion allele Vrn-A1b was present in only 1 of the western Canadian cultivars (Triticum aestivum L. 'Rescue') and 2 of its derivative chromosomal substitution lines. The intron deletion allele Vrn-A1c was not present in any line tested. Only 4 of the western Canadian spring wheat cultivars tested here carry the recessive vrn-A1 allele. The dominant allele of Vrn-B1 was detected in 20 cultivars/lines. Fourteen cultivars/lines had dominant alleles of Vrn-A1a and Vrn-B1 in combination. All cultivars/lines carried the recessive allele for Vrn-D1. The predominance of the dominant allele Vrn-A1a in Canadian spring wheat appears to be due to the allele's vernalization insensitivity, which confers earliness under nonvernalizing growing conditions. Wheat breeders in western Canada have incorporated the Vrn-A1a allele into spring wheats mainly by selecting for early genotypes for a short growing season, thereby avoiding early and late season frosts. For the development of early maturing cultivars with high yield potential, different combinations of Vrn alleles may be incorporated into spring wheat breeding programs in western Canada.     

Retrotransposon Insertional Polymorphism in Iranian Bread Wheat Cultivars and Breeding Lines Revealed by IRAP and REMAP Markers

Inter-retrotransposon amplified polymorphisms (IRAPs) and retrotransposon-microsatellite amplified polymorphisms (REMAPs) were used to detect retrotransposon integration events and genetic diversity in 101 Iranian bread wheat (Triticum aestivum L.) cultivars and breeding lines. The 9 IRAP primers amplified 128 loci, and 20 REMAP primers amplified 263 loci. Percentage of polymorphic loci, average expected heterozygosity, number of effective alleles, and Shannon’s information index for the REMAP markers were slightly higher than those for the IRAP markers. The same estimated parameters calculated for native and nonnative retrotransposons were not considerably different. A Mantel test between IRAP and REMAP cophenetic matrices evidenced no significant correlation. Cluster analysis based on the Dice similarity coefficient and complete linkage algorithm using IRAP+REMAP loci identified five groups among the genotypes studied that could be applied as crossing parents in T. aestivum breeding programs.     

Assessment of genetic diversity in Brazilian barley using SSR markers

Barley is a major cereal grown widely and used in several food products, beverage production and animal fodder. Genetic diversity is a key component in breeding programs. We have analyzed the genetic diversity of barley accessions using microsatellite markers. The accessions were composed of wild and domesticated barley representing genotypes from six countries and three breeding programs in Brazil. A total of 280 alleles were detected, 36 unique to Brazilian barley. The marker Bmag120 showed the greatest polymorphism information content (PIC), with the highest mean value found on chromosome three, and the lowest on chromosomes four and six. The wild accessions presented the highest diversity followed by the foreign genotypes. Genetic analysis was performed using Principal Coordinates Analysis, UPGMA clustering, and Bayesian clustering analysis implemented in Structure. All results obtained by the different methods were similar. Loss of genetic diversity has occurred in Brazilian genotypes. The number of alleles detected in genotypes released in 1980s was higher, whereas most of the cultivars released thereafter showed lower PIC and clustered in separate subgroups from the older cultivars. The use of a more diverse panel of genotypes should be considered in order to exploit novel alleles in Brazilian barley breeding programs.     

Trends in genetic diversity change of spring bread wheat cultivars released in Russia in 1929-2003

Using genealogy analysis, we studied genetic diversity of 340 cultivars of spring bread wheat that were released on the territory of Russia in 1929-2003. Trends in the temporal change of genetic diversity were inferred from analysis of a set of n x m matrices, where n is the number of the released cultivars and m is the number of original ancestors. The pool of original ancestors of the spring bread wheat cultivars for the total period of study included 255 landraces, of which 88 were from the former USSR and modern Russia. The original ancestors showed great differences in their presence in the cultivar sets examined and, consequently, in their importance for the gene pool of Russian spring wheats. The distributions of contributions of dominant original ancestors to cultivar diversity were significantly different in different regions, indicating that the ancestors were specific for the cultivation conditions. During the last 75 years, the genetic diversity of the spring bread wheat cultivars has been increasing owing to the wide use of foreign material in Russian breeding programs. However, our analysis showed that about 60 landraces, including the Russian ones, were lost during the studied time period. The lost part makes up 35% of the gene pool of the Russian original ancestors. It is reasonable to assume that the lost landraces carried a gene complex f or adaptation to specific Russian environments. Specificity of the contributions of the original ancestors in the sets of cultivars produced in different breeding centers was established. A comparative analysis of genetic similarity of cultivars was carried out using coefficients of parentage. Significant differences in this parameter between breeding institutes and regions of cultivation were revealed.     

Comparison of genomic SSR and EST-SSR markers for estimating genetic diversity in cucumber

Thirteen genomic microsatellite (gSSR) and sixteen expressed sequence tag (EST)-SSR (eSSR) markers were compared to estimate genetic diversity among 29 cucumber (Cucumis sativus L.) accessions. gSSR markers detected mean 4.46 alleles with a mean polymorphic information content (PIC) of 0.664, against eSSR markers with mean 3.38 alleles and a mean PIC of 0.397. gSSRs amplified more null alleles than eSSRs. Genetic diversity within the accession set was estimated by construction of dendrograms using gSSR or eSSR data. There was a clear consistency between gSSR and eSSR trees in terms of positioning of most cucumber germplasms. gSSR markers could separate various types of cucumber germplasms on the whole, although clustering of some accessions was not based on their geographical origins in eSSR tree. eSSR markers identified an independent sub-cluster containing five accessions resistant to downy mildew, suggesting a probable relationship between eSSRs and disease-resistance trait in cucumber. The Mantel test between gSSR and eSSR matrices revealed a good fit correlation (r = 0.836). The general dendrogram constructed using the combined data of gSSRs and eSSRs was similar to those obtained separately with each marker.     

Analysis of genetic diversity and relationships among waxy maize inbred lines in Korea using SSR markers

Information regarding the genetic diversity and genetic relationships among elite inbred lines is necessary to improve new cultivars in maize breeding programs. In this study, genetic diversity and genetic relationships were investigated among 84 waxy maize inbred lines using 50 SSR markers. A total of 269 alleles were identified at all the loci with an average of 5.38 and a range between 2 and 13 alleles per locus. The gene diversity values varied from 0.383 to 0.923 with an average of 0.641. The cluster tree generated using the described SSR markers recognized two major groups at 32% genetic similarity. Group I included 33 inbred lines while group II included 51 inbred lines. The clustering patterns of most of the waxy maize inbred lines did not clearly agree with their source, pedigree or geographic location. The average GS among all inbred lines was 35.7 ± 10.8. Analysis of waxy maize inbred lines collected from Korea and China at 50 SSR loci revealed higher values of average number of alleles (4.9) and gene diversity (0.638) in Korean inbred lines as compared to Chinese inbred lines (3.5 and 0.563, respectively). The information obtained from the present studies would be very useful for maize breeding programs in Korea.     

黄淮麦区小麦品种(系)的ISSR位点遗传多样性分析

选用11个ISSR引物,对黄淮麦区96个小麦推广品种（系）进行遗传多样性分析。共检测到96个多态性位点,每个引物多态性位点数平均为8．7个,变幅为3～23个;ISSR引物的多态性信息含量PIC变幅为0．601～0．941,平均0．791,表明ISSR具有较强的品种间区分能力,是研究小麦种质资源遗传多样性的有效分子标记技术之一。96个品种（系）间,Nei’s遗传相似系数变化范围为0．53～0．91,平均为0．60,品种间遗传相似性变幅较大,表明黄淮麦区不同小麦品种（系）间存在着不同程度的遗传多样性差异。根据品种间遗传相似系数聚类,96份材料被聚成8大类群,共14个亚类,类群与系谱和原产地无关。     

Analysis of diversity of Russian and Ukrainian bread wheat (Triticum aestivum L.) cultivars for high-molecular-weight glutenin subunits

The allelic diversity of high-moleculat-weght glutenin subunits (H WIGS) in Russian and Ukrainian bread wheat cultivars was analyzed. The diversity of spring wheat cultivars for alleles of the Glu-1 loci is characterized by medium values of the polymorphism index (polymorphism information content, PlC), and in winter wheats it varies from high at the Glu-A1 locus to low at the Glu-D1 locus. The spring and winter cultivars differ significantly in the frequencies of alleles of the glutenin loci. The combination of the Glu-A1b, Glu-B1c, and Glu-D1a alleles prevails among the spring cultivars, and the combination of the Glu-A1a, Glu-B1c, and Glu-D1d alleles prevails among the winter cultivars. The distribution of the Glu-1 alleles significantly depends on the moisture and heat supply in the region of origin of the cultivars. Drought resistance is associated with the Glu-D1a allele in the spring wheat and with the Glu-B1b allele in the winter wheat. The sources of the Glu-1 alleles were identified in the spring and wheat cultivars. The analysis of independence of the distribution of the spring and winter cultivars by the market classes and by the alleles of the HMWGS loci showed a highly significant association of the alleles of three Glu-1 loci with the market classes in foreign cultivars and independence or a weak association in the Russian and Ukrainian cultivars. This seems to be due to the absence of a statistically substantiated system of classification of the domestic cultivars on the basis of their quality.