Combined use of gliadins and SSRs to analyse the genetic variability of the Spanish collection of cultivated diploid wheat (Triticum monococcum L. ssp. monococcum)

This work studied the combined use of gliadins and SSRs to analyse inter- and intra-accession variability of the Spanish collection of cultivated einkorn (Triticum monococcum L. ssp. monococcum) maintained at the CRF-INIA. In general, gliadin loci presented higher discrimination power than SSRs, reflecting the high variability of the gliadins. The loci on chromosome 6A were the most polymorphic with similar PIC values for both marker systems, showing that these markers are very useful for genetic variability studies in wheat. The gliadin results indicated that the Spanish einkorn collection possessed high genetic diversity, being the differentiation large between varieties and small within them. Some associations between gliadin alleles and geographical and agro-morphological data were found. Agro-morphological relations were also observed in the clusters of the SSRs dendrogram. A high concordance was found between gliadins and SSRs for genotype identification. In addition, both systems provide complementary information to resolve the different cases of intra-accession variability not detected at the agro-morphological level, and to identify separately all the genotypes analysed. The combined use of both genetic markers is an excellent tool for genetic resource evaluation in addition to agro-morphological evaluation.     

Characterisation and Variation of Morphological Traits and Storage Proteins in Spanish Emmer Wheat Germplasm (<Emphasis Type="Italic">Triticum Dicoccon</Emphasis>)

Emmer wheat is hulled wheat that was wide cultivated in Spain at the past. Actually, the most of this germplasm is conserved in Germplasm Banks, and only two small populations have been found in Asturias (North of Spain) in a recent collecting mission. In this work, a collection of 31 Spanish emmer lines developed from identical number of accessions of two Germplasm Banks was analysed for morphological spike traits and seed storage protein composition. Up to seven different botanical varieties were detected, which suggest the presence of a wide diversity, although lower than the historically described 10 botanical varieties. At level of seed storage proteins, the lines showed a high diversity, although the new alleles were present with low frequency in materials with scarce agronomic interest for the farmers (var. atratum, var. lagascae or var. pycnurum). This last circumstance could translate in a lost of variability by genetic drift.     

Analysis of Genetic Variability in a Sample of the Durum Wheat (Triticum durum Desf.) Spanish Collection Based on Gliadin Markers

In this work gliadin proteins were used to analyse the genetic variability in a sample of the durum wheat Spanish collection conserved at the CRF-INIA. In total 38 different alleles were identified at the loci Gli-A1, Gli-A3, Gli-B5, Gli-B1, Gli-A2 and Gli-B2. All the gliadin loci were polymorphic, possessed large genetic diversity and small and large differentiation within and between varieties, respectively. The Gli-A2 and Gli-B2 loci were the most polymorphic, the most fixed within varieties and the most useful to distinguish among varieties. Alternatively, Gli-B1 locus presented the least genetic variability out of the four main loci Gli-A1, Gli-B1, Gli-A2 and Gli-B2. The Gli-B1 alleles coding for the gliadin γ-45, associated with good quality, had an accumulated frequency of 69.7%, showing that the Spanish germplasm could be a good source for breeding quality. The Spanish landraces studied showed new gliadin alleles not catalogued so far. These new alleles might be associated with specific Spanish environment factors. The large number of new alleles identified also indicates that durum wheat Spanish germplasm is rather unique.     

Variation in Spanish cultivated einkorn wheat (<Emphasis Type="Italic">Triticum monococcum</Emphasis> L. ssp. <Emphasis Type="Italic">monococcum</Emphasis>) as determined by morphological traits and waxy proteins

A collection of 29 lines of Spanish einkorn wheat (Triticum monococcum L. ssp. monococcum) were analysed for morphological traits and waxy proteins. Four different botanical varieties were detected in the collection on the basis of the morphological traits. For the waxy proteins, no polymorphism has been described in previous works in wheat diploid species. In the current materials, two different alleles were found, namely Wx-A ^m 1a and Wx-A ^m 1a′. The allele Wx-A ^m 1a′ that presents less mobility than the Wx-A ^m 1a, has not been previously described and was only found in one line.     

Protein Heterogeneity in European Wheat Landraces and Obsolete Cultivars: Additional Information II

The endosperm storage protein of 46 European wheat (Triticum aestivum L.) landraces and obsolete cultivars have been fractionated by SDS-PAGE to determine the composition of high molecular weight glutenin subunits (HMW-GS) composition. It has been discovered that about 46% of the wheats were heterogeneous, comprising 2–11 different glutenin profiles. Eighteen of them were observed to be homogeneous. A total of 13 HMW-GS alleles, including 3 at the Glu-A1, 8 at the Glu-B1, and 3 at the Glu-D1 loci were revealed. HMW-GS null controlled by locus Glu-A1, subunits 7 + 8 by Glu-B1, and 2 + 12 by Glu-D1 predominated. However low frequented alleles such as 17 + 18, 20, 6, and 7 were observed. Furthermore, other new alleles encoding HMW-GS at the locus Glu-B1 have been found in one of France cultivar (Saumur d’Automne). The glutenin-based quality score ranged from 4 to 10.     

Microsatellite mapping of genes for branched spike and soft glumes in Triticum monococcum L.

The locus responsible for branched spike in a branched spike mutant of Triticum monococcum L. (2n = 2x = 14, A^mA^m genome) and soft glume in Triticum sinskajae Filat. et Kurkiev (2n = 2x = 14, A^mA^m genome) were mapped by genotyping F₂ populations using microsatellite markers. Phenotypic analysis in the cross T. sinskajae PI 418587/a branched spike mutant KT3-24 confirmed that both characters were under control of a recessive allele at a single locus, and they were linked with 26.6 cM. The branched head in T. monococcum (bh ^m ) locus was located on chromosome 2A^mS and the marker Xgwm122 flanked the bh ^m gene distally. Soft glume locus in T. sinskajae was allelic to the soft glume (sog) locus in mm09, a soft glume mutant of T. monococcum. The sog locus was linked with Xwmc644 distally. In the F₂ hybrids of T. monococcum #252/PI 418587 and T. monococcum KT 3-21/PI 418587, sog was linked with Xgwm71. The gene fg which determines a false glume was also located on chromosome 2A^mS and the recombination between sog and fg (1.6 cM) was obtained in F₂ hybrid of KT 3-21/PI 418587.     

六倍体小黑麦品种资源Glu-1位点的多态性

利用SDS-PAGE技术分析了我国新疆101份和波兰11份六倍体小黑麦品种资源高分子量谷蛋白亚基(HMW-GS)组成,共检测到17种高分子量谷蛋白亚基,其中Glu-A1位点编码的HMW-GS有3种变异类型,即1(a)、2*(b)和Null(c),Glu-B1位点编码的HMW-GS有8种变异类型,即7(a)、7+8(b)、7+9(c)、6+8(d)、20(e)、13+19(g)、7+18(r)和6.8+20y(s),Glu-R1位点编码的HMW-GS有6种变异类型,即1r+4r(a)、2r+6.5r(b)、6r+13r(c)、2r+9r(d)、6.5r(e)和0.8r+6r(f)。这些小黑麦品种的HMW-GS组成以Null(c)、7+18(r)和6r+13r(c)为主,分别占58.93%、67.90%和58.00%。在112份供试材料中共检测到30种HMW-GS组合变异类型,其中[Null,7+18,6r+13r(c,r,c)]和[2*,7+18,6r+13r(b,r,c)]出现频率较高,分别为16.91%和16.02%,其他类型组合出现频率较低,个别材料具有少见的特殊亚基组合,如[2*,7+18,2r+9r(b,r,d)]、[2*,6.8+20y,2r+6.5r(b,s,b)]等类型。分析2个地域品种的遗传多样性,发现新疆品种的遗传变异范围小于波兰品种,波兰品种在Glu-1位点上的遗传变异更丰富。结果还显示,在六倍体小黑麦的人工进化过程中Glu-B1位点发生了很大变异,产生了小黑麦特有的7+18(r)和6.8+20y(s)亚基,而且频率很高,为小麦品质改良提供了丰富的基因资源。     

Genetic diversity of HMW glutenin subunit in Chinese common wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) landraces from Hubei province

The high molecular weight (HMW) glutenin subunit composition of 111 common landraces of bread wheat collected from Hubei province, China has been determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Ninety six of the accessions were homogeneous for HMW glutenin subunit composition and 15 were heterogeneous. For the Glu-1 loci, 16 alleles were detected, 3 at the Glu-A1locus, 9 at the Glu-B1and 4 at the Glu-D1. Three novel alleles were identified, two at the Glu-B1 and one at the Glu-D1locus. Combination of these 16 alleles resulted in 14 different HMW subunit patterns. The distribution of HMW glutenin subunit alleles in a subset of 105 of the 111 accessions representing six populations was assessed both at the individual population and whole population levels. The results demonstrated that the distribution of allelic patterns varied among populations. Taken together, 62.5% of the alleles detected were considered to be rare alleles while the Glu-A1c (null), Glu-B1b (1Bx7 + 1By8) and Glu-D1a (1Dx2 + 1Dy12) alleles were found most frequently in the six populations. The subset exhibited relatively high genetic diversity (A = 5.33, P = 1.00, Ae = 1.352 and He = 0.238) with 81.5% of the diversity being within populations and 18.5% between populations.     

(英文)

用SDS-PAGE方法测定了我国10个小麦主产省份171份小麦品种和高代品系的高分子量麦谷蛋白亚基（HMW-GS）组成。鉴定出18种HMW-GS，40种HMW-GS组成形式，其中20种亚基其组成形式只在一个品种（系）中出现。Glu-A1位点亚基1和Null出现最多，Glu-B1位点7＋8和7＋9亚基对占绝对优势，Glu-D1位点2＋12亚基对出现频率最高。Null、7＋9、2＋12、Null，7＋8，2＋12，1、7＋8，2＋12，1、7＋9、2＋12等亚基组成形成出现频率最高，占分析品种的49.71％。与前人研究相比，新育成品种HMW-GS亚基组成发生了明显变化，面包优质亚基（对）1、5＋10出现的频率显著升高，亚基多态性增加，组成形式明显改善，这些对于品质改良和品种选育是非常有利的，新育成品种Glu-1品质评分已超过7。尽管个别品种亚基组成好，品质优良，但总体上看，我国小麦品种与其它国家相比品质还存在一定差距，提高5＋10、17＋18等优质亚基的频率是改善我国小麦面包品质的重要措施。     

(英文)

对来源于美、中、俄及埃塞阿比亚等22个国家的142份硬粒小麦材料的种子贮藏蛋白位点及遗传变异进行了研究。供试的硬粒小麦(Triticum durum Desf )材料共检测出37条醇溶蛋白条带，无1条带纹为所有材料共有，多态性达到100％，说明硬粒小麦具有丰富的醇溶蛋白等位变异。聚类分析将142份供试材料分为3个大类，材料间遗传差异大小在不同的国家有所不同，表明醇溶蛋白带型与地理来源有一定关系。高分子量谷蛋白电泳共分离出14种亚基和15种亚基组合，但是优质亚基所占比例不高，这可能是因为硬粒小麦加工用途的特殊性，使得多年的育种并未太多改变硬粒小麦高分子量谷蛋白亚基等位变异的频率，促成优质亚基的累计。     

The effect of soil-climate conditions on yielding parameters,chemical composition and baking quality of ancient wheat species Triticum monococcum L., Triticum dicoccum Schrank and Triticum spelt L. in comparison with modern Triticum aestivum L.

Triticum monococcum L., Triticum dicoccum Schrank and Triticum spelt L. nowadays offer an alternative to Triticum aestivum L. We analyzed grain and straw yield, yielding parameters, chemical composition and bakery quality of these species and compared them with modern T. aestivum at three sites with different soil-climate conditions. The average grain yield varied from 0.41 t ha^?1 (T. monococcum) to 5.17 t ha^?1 (T. aestivum), straw yield varied from 1.50 t ha^?1 (T. dicoccum) to 5.83 t ha^?1 (T. aestivum). The yielding parameters and chemical composition of the grain were significantly influenced by soil-climate conditions and wheat species. The highest average crude protein content was recorded in T. spelta (20.55%), while the lowest in T. aestivum (11.20%). The Zeleny’s sedimentation test ranged from 9.0 ml (T. monococcum) to 34.5 ml (T. aestivum) and the value of the Gluten index varied from 7.45 (T. dicoccum) to 89.75 (T. aestivum). According to the results, ancient wheat species provides lower grain and straw yields, higher protein content and mineral concentrations. Concentration of proteins and grain’s baking quality strongly depends on wheat species and soil-climate conditions.     

Genetic diversity of HMW glutenin subunits in diploid,tetraploid and hexaploid <Emphasis Type="Italic">Triticum</Emphasis> species

The genetic variations of high-molecular-weight (HMW) glutenin subunits in 1051 accessions of 13 Triticum subspecies were investigated using sodium dodecyl sulfate polyacrylamide-gel electrophoresis. A total of 37 alleles were detected, resulting in 117 different allele combinations, among which 20, 68 and 29 combinations were observed in diploid, tetraploid and hexaploid wheats, respectively. Abundance and frequency of allele and combinations in tetraploid wheats were higher than these in hexaploid wheats. Allele Glu-A1c was the most frequent subunit at Glu-A1 locus in tetraploid and hexaploid wheats. Consequently, the results also suggested that the higher variations occurred at Glu-B1 locus compared to Glu-A1 and Glu-D1. Therefore, carthlicum wheat possessing the allele 1Ay could be presumed a special evolutional approach distinguished from other tetraploid species. Furthermore, this provides a convenient approach of induction of the 1Ay to common wheat through direct cross with carthlicum wheat. Alleles Glu-B1c and Glu-B1i generally absent in tetraploid wheats were also found in tetraploid wheats. Our results implied that tetraploid and hexaploid wheats were distinguished in dendrogram, whereas carthlicum and spelta wheats and however displayed the unique performance. In addition, founder effect, no-randomness of diploidization, mutation and artificial selection could cause allele distribution of HMW-GS in Triticum. All alleles of HMW-GS in Triticum could be further utilized through hybrid in the quality improvement of common wheat.     

Assessment of genetic variability in hexaploid wheat landraces of Pakistan based on polymorphism for HMW glutenin subunits

Summary Sixty hexaploid wheat landraces collected from five regions of Pakistan were assessed for genetic variability in terms of high molecular weight (HMW) glutenin subunits as revealed by SDS-PAGE. The germplasm appeared to be diverse and unique on the basis of HMW glutenin subunit compositions. Out of 24 alleles detected at all the Glu-1 loci, four belonged to Glu-A1, 12 to Glu-B1 and eight to Glu-D1 locus. The number of novel HMW glutenin subunits detected were 1, 4 and 6 at the three loci (Glu-A1, Glu-B1, Glu-D1), respectively. The frequency distribution patterns of 24 allelic variants detected at the three Glu-1 loci in 1080 samples analysed for 60 accessions were determined both on the basis of individual accessions and on the basis of regions (accessions pooled across the regions). One allele (null) at the Glu-A1 locus, three alleles (17+18, 7+8, 14) at the Glu-B1 locus and, two alleles (2+12 and 2^**+12) at the Glu-D1 locus were found most frequently distributed in the 60 populations. Maximum variation was observed in the Baluchistan and Gilgit regions of Pakistan in terms of distribution of novel Glu-1 alleles. A higher gene diversity was observed between the populations as compared to the gene diversity within the populations while, a reverse pattern of gene diversity was observed when populations were pooled across the regions (higher within the regions than between the regions). A data base has been generated in this study which could be expanded and usefully exploited for cultivar development or management of gene bank accessions.     

Genetic Diversity of Glutenin Protein Subunits Composition in Durum Wheat Landraces [Triticum turgidum ssp. turgidum Convar. durum (Desf.) MacKey] from the Mediterranean Basin

The genetic diversity of high and low molecular weight glutenin subunits of 63 durum wheat landraces from different geographical regions in the Mediterranean Basin was studied using SDS-PAGE. Great variability in glutenin composition was found, with 42 high and low molecular weight glutenin haplotypes, 20 allele combinations at the HMW-GS loci, and 18 at the LMW-GS. All five possible LMW models were detected in all Mediterranean regions. Rare alleles were found at Glu-B1 locus in high frequencies and a priori related alleles to grain quality were also observed. Global genetic diversity index was relatively high (0.67); it ranged from 0.33 to 0.66. Cluster analysis on the frequency patterns of origins grouped genotypes following a geographical structure. Rogers’ distance coefficient on frequency pattern for each region of origin showed two germplasm pools with distinct quality profiles, where South West Asian landraces were very different from the landraces of other Mediterranean areas. The relationship between different regions of origin is discussed and two possible ways of introduction of wheat in the Iberian Peninsula (N Africa and SE Europe) are hypothesized. The use of Mediterranean durum wheat landraces as source of genetic variability for grain quality improvement is highly recommended.     

Genetic variability in bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) of Pakistan based on polymorphism for high molecular weight glutenin subunits

Variation in bread wheat including pre and post green revolutions varieties of Pakistan along with landraces was investigated for high molecular weight Glutenin subunits (HMW Gs) encoded at three genes (Glu-A1, Glu-B1, Glu-D1) with SDS-PAGE. The germplasm was diverse and unique on the basis of HMW Gs compositions and out of 14 alleles detected at all the Glu-1 loci, three belonged to Glu-A1, nine to Glu-B1 and two to Glu-D1 locus. High variation was observed in the landraces and higher gene diversity was observed between the populations as compared to the gene diversity within populations, whereas a reverse pattern of gene diversity was observed when populations were pooled across the region (higher within the regions than between the regions). A lack of relationship between the HMW Gs diversity and the altitude of collection site was observed. A data base has been generated in this study which could be expanded/exploited for cultivar development or management of gene bank.     

黄土丘陵沟壑区红砂灌丛土壤种子库及其自然更新潜力评估

[目的]对黄土丘陵沟壑区红砂灌丛植被土壤种子库的特征及红砂灌丛植被自然更新潜力进行分析和评估,以说明红砂灌丛土壤种子库在植被恢复与重建中的重要作用。[方法]采用土壤种子库"萌发法"(每个地段10个2m×2m样方内分表土层0—2cm和2—5cm土层采集土样)及野外植被调查方法,对黄土丘陵沟壑不同坡位红砂灌丛土壤种子库进行研究。[结果]黄土丘陵沟壑区红砂灌丛植被土壤种子库发芽试验共观察到的4 251株幼苗,分属于9个物种。土壤种子库密度在100~1 000粒/m2,物种数在0.8~1.2种/0.01m2,下坡段的土壤种子库平均种子密度和平均物种数均比上坡段高。3个坡段的2个层次的平均种子密度和平均物种数均随土层加深而减小。3个坡段土壤种子库和地上植被的组成物种多为草本植物和红砂灌丛,地上植被与其土壤种子库的密度及物种数均呈不显著相关;物种组成的Sorensen相似性指数较高,均达到0.60以上,且土壤种子库比地上植被具有更高的物种丰富度。[结论]黄土丘陵沟壑区红砂灌丛具有依靠土壤种子库实现自然更新的潜力,但由于物种组成种类较少,现存灌丛植被一旦遭到破坏,仅靠土壤种子库恢复现存植被是困难的,需要在对自然恢复潜力评估的基础上,积极采取人工保护促进自然恢复的策略。     

DNA Fingerprinting and Genetic Characterization of Anatolian Triticum spp. using AFLP Markers

In this study, genetic analysis of Triticum spp. was carried out using AFLP markers. Six AFLP selective combinations were scored as presence and absence of bands for all the individual samples obtained from a single seed of each accession (70 accessions); T. baeoticum (21), T. monococcum (5), T. urartu (16), T. araraticum (7), T. dicoccoides (16) and T. dicoccon (5), resulting in 506 polymorphic AFLP bands. The phylogenetic tree showed two major clusters; one was composed of T. monococcum (AA) and T. baeoticum (AA), and the other cluster included T. araraticum (AAGG), T. dicoccon (AABB), T. dicoccoides (AABB), and T. urartu (AA). T. urartu, although having a diploid AA genome, did not cluster with other A genome diploids such as T. monococcum and T. baeoticum; instead it clustered together with the tetraploid species, confirming that T. urartu is the A genome progenitor. The extent of variations within and among species is discussed.     

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.     

Variability and Genetic Diversity for Gliadins in Natural Populations of Hordeum Chilense Roem. et Schult.

Hordeum chilense Roem. et Schult. is wild diploid barley that has been used for the synthesis of the new cereal, tritordeum (× Tritordeum Ascherson et Graebner). This species is distributed in Chile and Argentina where it is possible to find in natural populations. In this work, the gliadin fraction of the seed storage protein have been used for evaluating the genetic diversity in nine natural populations of H. chilense. The total diversity (Ht) in the nine populations was 0.879 (ranging from 0.926 for the ω-gliadins to 0.795 for the β-gliadins). Data showed that the 71.3% of this genetic diversity was between populations, whereas the 28.7% was within populations. This species is considered as self-pollinating, where gene flow within populations was mainly attributed to seed dispersal because of these materials appear distributed in cattle zones where the pasturing could be an efficient mechanism of dispersion.     

Population diversity of the Vicia sativa agg. (Fabaceae) in the flora of the former USSR deduced from RAPD and seed protein analyses

The genetic diversity of 58 wild and weedy populations representing taxa within the V. sativa aggregate from the former USSR, 4 cultivars of V. sativa, 2 accessions of V. cordata and 3 accessions of V. macrocarpa from Mediterranean countries were analysed using randomly amplified DNA fragments (RAPDs) and seed protein electrophoresis (SDS-PAGE). Interspecific variation between taxa in the V. sativa aggregate could readily be detected using both techniques. RAPDs and seed protein patterns were found to be an effective means of identifying accessions that cannot be identified clearly by morphological criteria alone. RAPD and seed protein analysis revealed a clear relationship between observed genetic variation of populations and their geographical distribution. Populations from each region had their own gene pools. Geographical variation was detected in V. segetalis. The degree of genetic divergence between local populations was usually related to proximity. In several locations where wild and weedy populations of different V. sativa agg. taxa grow sympatrically, intermediate forms could be detected at the DNA and protein levels. Both RAPD and seed protein analysis support the view that the V. sativa aggregate consists of 8 taxa warranting recognition at the species level. Several species in this aggregate are evolving intra-specific groups which can readily be detected at the molecular level.