Genetic polymorphisms at Gli-Dt gliadin loci in Aegilops tauschii as revealed by acid polyacrylamide gel and capillary electrophoresis

Gliadin variation at Gli‐D^t1 and Gli‐D^t2 loci in 198 Aegilops tauschii accessions was studied by acid polyacrylamide gel electrophoresis (A‐PAGE) and capillary electrophoresis (CE). High genetic polymorphisms were found at both gliadin coding loci, revealing a total of 184 and 169 gliadin variants at the Gli‐D^t1 and Gli‐D^t2 loci, respectively. In particular, 12 gliadin blocks encoded by different alleles were apparently expressed and readily identified in six synthetic hexaploids produced by hybridization between Triticum durum and Ae. tauschii accessions. Compared with Ae. tauschii ssp. eusquarrosa, the gliadin profile of the D genome in Ae. tauschii ssp. strangulata more resembles that of T. aestivum, supporting the view that the subspecies strangulata is the most likely progenitor of bread wheat. Capillary electrophoresis analysis showed that the method is capable of separating and characterizing gliadins with speed, in high resolution using small sample amounts, and is well‐suited to detect protein alleles and to identify desirable genotypes in wheat quality improvement.     

Allelic variation and genetic diversity of HMW glutenin subunits in Chinese wheat (Triticum aestivum L.) landraces and commercial cultivars

Wheat landraces have abundant genetic variation at the Glu-1 loci, which is desirable germplasms for genetic enhancement of modern wheat varieties, especially for quality improvement. In the current study, we analyzed the allelic variations of the Glu-1 loci of 597 landraces and 926 commercial wheat varieties from the four major wheat-growing regions in China using SDS-PAGE. As results, alleles Null, 7+8, and 2+12 were the dominant HMW-GSs in wheat landraces. Compared to landraces, the commercial varieties contain higher frequencies of high-quality alleles, including 1, 7+9, 14+15 and 5+10. The genetic diversity of the four commercial wheat populations (alleles per locus (A) = 7.33, percent polymorphic loci (P) = 1.00, effective number of alleles per locus (Ae) = 2.347 and expected heterozygosity (He) = 0.563) was significantly higher than that of the landraces population, with the highest genetic diversity found in the Southwestern Winter Wheat Region population. The genetic diversity of HMW-GS is mainly present within the landraces and commercial wheat populations instead of between populations. The landraces were rich in rare subunits or alleles may provide germplasm resources for improving the quality of modern wheat.     

Genome discrimination and chromosome pairing in the Hordeum chilense × Aegilops tauschii amphiploid

Tritordeum (X Tritordeum Ascherson et Graebner) is a synthetic amphiploid belonging to the Triticeae tribe, which resulted from crosses between Hordeum chilense and wheat. It presents useful agronomic traits that could be transferred to wheat, widening its genetic basis. In situ hybridisation with total genomic DNA from H. chilense and cloned, repetitive DNA sequences (pTa71 and pAs1) probes were used to discriminate the parental origin of all chromosomes, to analyse the chromosome pairing and to identify the chromosomes in pollen mother cells (PMCs) at metaphase I of the tritordeum line HT251 (H^chH^chDD, 2n = 4x = 28). The H. chilense total genomic DNA and the ribosomal sequence pTa71 probes, allowed the unequivocal discrimination of the 14 chromosomes of H^ch genome-origin and the 14 chromosomes of D genome-origin. Chromosome pairing analysis revealed meiotic irregularities such as reduced percentage of PMCs with complete homologous pairing, high frequency of univalents, most of H. chilense-origin and a reduced frequency of intragenomic multivalents from both genomes. The H. chilense genome revealed high meiotic instability. After individual chromosome identification at metaphase I with the pAs1 probe, we found the occurrence of pairing between chromosomes of different homoeology groups. The possible interest of the tetraploid tritordeum in the improvement of other Triticeae species is also discussed.     

Disc polyacrylamide gel electrophoresis of pollen proteins in the oil palm (Elaeis)

Summary A preliminary screening of proteins and isozymes in the oil palm was investigated with a view to using the data in discriminating oil palm fruit forms. Protein and enzyme staining was carried out using pollen tissue. Repeatable bands of proteins which showed reproducable variation in banding patterns were obtained. Low percentage similarities between the groups compared for protein banding patterns were obtained. A single enzyme esterase extracted from pollen tissue exhibited repeatable variation between species only. It was intended to obtain information that would supplement morphological markers through electrophoretic data in the identification of different fruit forms of the oil palm.     

Development of diversity array technology (DArT) markers for assessment of population structure and diversity in Aegilops tauschii

Aegilops tauschii Coss. is the D-genome donor to hexaploid bread wheat (Triticum aestivum) and is the most promising wild species as a genetic resource for wheat breeding. To study the population structure and diversity of 81 Ae. tauschii accessions collected from various regions of its geographical distribution, the genomic representation of these lines were used to develop a diversity array technology (DArT) marker array. This Ae. tauschii array and a previously developed DArT wheat array were used to scan the genomes of the 81 accessions. Out of 7500 markers (5500 wheat and 2000 Ae. tauschii), 4449 were polymorphic (3776 wheat and 673 Ae. tauschii). Phylogenetic and population structure studies revealed that the accessions could be divided into three groups. The two Ae. tauschii subspecies could also be separately clustered, suggesting that the current taxonomy might be valid. DArT markers are effective to detect very small polymorphisms. The information obtained about Ae. tauschii in the current study could be useful for wheat breeding. In addition, the new DArT array from this Ae. tauschii population is expected to be an effective tool for hexaploid wheat studies.     

A proposed method of polyacrylamide gel electrophoresis in acid environments applied to gliadins of wheat grains

Summary In this paper we describe the method of wheat gliadin electrophoresis in use at RIVRO, Wageningen. It differs from other techniques mainly by the application of an alternative buffer system, making it possible to polymerise the gels in a buffered alkaline environment and to perform the run at pH 3.1 without extensive buffer changing steps. Advantages are a greater gel reproducibility and the ease of gel handling. Furthermore, a rationalised protein extraction procedure, a cheap shaking system for staining baths and a better (slower moving) tracking dye are described.     

The use of polyacrylamide gradient gel electrophoresis to identify variation in isozymes as markers for Lactuca species and resistance to the lettuce root aphid Pemphigus bursarius

Summary This paper describes simple, routine polyacrylamide gradient gel electrophoresis of allozyme patterns surveyed in wild populations of four Lactuca species, obtained from the Wellesbourne Vegetable Gene Bank. Using multivariate statistical techniques, four allozyme bands were identified, which distinguished the four species, and an additional two bands, which were related to the resistance of Lactuca species to the lettuce root aphid.     

Genetic diversity of European spelt wheat (Triticum aestivum ssp. spelta L. em. Thell.) revealed by glutenin subunit variations at the Glu-1 and Glu-3 loci

Summary High and low molecular weight glutenin subunit (HMW-GS and LMW-GS) compositions of 270 European spelts, 15 Iranian spelts and 25 bread wheat cultivars were analyzed by one- and two-dimensional gel electrophoresis. The results revealed a total of 22 HMW-GS alleles (4 at Glu-A1, 11 at Glu-B1 and 7 at Glu-D1) and 32 allele combinations among the three Glu-1 loci. Two major genotypes of HMW-GS: 1, 13+16, 2+12 and 1, 6.1+22.1, 2+12 were found to occur in Central European spelt wheat cultivars and landraces at higher frequencies of 35 and 28%, respectively. The Glu-B1 locus displayed the greatest variation and genetic diversity index (H) was 0.69 whereas Glu-A1 and Glu-D1 showed H index values of 0.26 and 0.19, respectively. The dendrogram constructed by HMW and LMW glutenin subunit bands revealed that European spelts form a separated cluster from common wheat suggesting that spelt and common wheat form distinct groups. In addition, all 15 Iranian spelt land variety accessions differed from European spelts and possessed similar HMW-GS alleles to common wheat. Because of a wider polymorphism Central European spelt wheats are an important genetic reserviour for improving common wheat quality. Both authors contributed equally to this work     

SDS-PAGE of the high-molecular-weight subunits of wheat glutenin and the characterization of 1R (1B) substitution and 1BL/1RS translocation lines

Summary The high-molecular-weight subunits of glutenin from wheat 1R(1B) substitution and 1BL/1RS translocation lines were fractionated by SDS-PAGE. Two new subunits denoted R₁ and R₂ were characterized in 1R(1B) substitution, but not in 1BL/1RS translocation lines. R₁ and R₂ were proved to be rye proteins by 2d electrophoresis (NEPHGE x SDS-PAGE).In contrast to literature citations it was demonstrated that the cultivar Winnetou is a 1R(1B) substitution line and the cultivars Clement and Mildress both are 1BL/1RS translocation lines.     

Identification of cultivars of pasture legumes (Centrosema macrocarpum,C. pubescens and C. sp.n.) by acid polyacrylamide gel electrophoresis of cotyledons storage proteins

Summary An electrophoretic procedure was developed for storage proteins which can discriminate cultivars of forage legumes, Centrosema macrocarpum, C. pubescens and C. sp.n. Proteins extracted from cotyledons were separated in 10% polyacrylamide gel containing 10% sucrose. Electrophoretic patterns are presented to illustrate the results that can be obtained with the procedure described. Cultivars of all three species were distinguishable based upon the variation in their acid soluble seed proteins.Joint publiccation of the Department of Food Science, University of Manitoba (No 101) and Centro Internacional de Agricultura Tropical-CIAT.     

Random amplified polymorphic DNA (RAPD) variation in Fagopyrum tataricum Gaertn. accessions from China and the Himalayan region

The diversity among 52 landraces and cultivars of tartary buckwheat (Fagopyrum tataricum Gaertn.) and one accession of its wild ancestor, F. tataricum ssp. potanini Batalin, from diverse geographic origins was examined using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) markers. Eighteen primers produced a total of 240 fragments, of which 153 (63.75%) were monomorphic and 87 (36.25%) polymorphic bands. UPGMA-based pairwise Jaccard’s coefficient of similarity was used to deduce the relationships among 53 genetically diverse accessions. The similarity between cultivated tartary buckwheat accessions ranged from 0.61 to 1.00. Four distinct clusters were formed which corresponded well with the geographic distribution of the tartary buckwheat. Nepalese accessions showed maximum diversity followed by Chinese accessions. Tartary buckwheat accessions from the Himalayan region of northwestern India revealed a narrow gene pool. The wild buckwheat accession did not group with any of the three cultivated tartary buckwheat groups, and formed its own single-entry group. Genetic similarity (0.59) of Chinese buckwheat accessions with the wild ancestor reaffirmed that cultivated tartary buckwheat originated in the Yunnan province of northwestern China. Consistent with some earlier reports, our study demonstrated the usefulness of the RAPD technique for the characterization of plant genetic resources and assessment of diversity between species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Predicting interspecific compatibilities in beans (Phaseolus) by seed protein electrophoresis

Summary Seed proteins of 17 wild species of Phaseolus were separated by electrophoresis on SDS polyacrylamide gels. There was very little variation of the protein pattern within most species, while considerable variation among species was evident. Relative interspecific similarities of protein patterns were estimated using Jaccard's similarity index, and a cluster analysis was performed on these values. The resultant dendrogram generally agreed with previous research on interspecific relationships in Phaseolus based on morphological characteristics, and also generally agrees with current information on interspecific similarities based on hybridization studies. Suggestions are made regarding interspecific hybridization among Phaseolus species which have not been attempted, but which may be possible based on cluster analysis. These hybrids include P. vulgaris × P. polystachyus, P. vulgaris × (P. polystachyus × P. metcalfei) and P. anisotrichos × P. angustissimus.Supported by the USDA Specific Cooperative Agreement to the University of Puerto Rico 458-7B30-0-203 Bean Improvement by Interspecific Crosses.     

Allelic variation, sequence determination and microsatellite screening at the XGWM261 locus in Chinese hexaploid wheat (Triticum aestivum) varieties

Wheat microsatellite XGWM261, due to its closely linked to the dwarfing gene Rht8, has been adopted as the diagnostic molecular marker of Rht8. Screening 408 Chinese and 98 exotic varieties showed 13 allele variants in locus of XGWM261, with 6 alleles only to be found in Chinese varieties and 2 only in exotic varieties, respectively. Sequencing results of the 13 alleles revealed their absolute fragment sizes with 216, 212, 210, 206, 204, 202, 200, 196, 194, 192, 190, 174, and 164 bp, respectively. Allelic distribution analysis showed that the 204, 192, 174, and 164 bp alleles were prevailing in Chinese varieties, and the diagnostic 192 bp allele to Rht8 had a very high percentage in the Yellow and Huai River Valleys Facultative Wheat Zone than in the Northern Winter Wheat Zone in China. The GT → AC substitution at position 35 was found in 216, 200, and 174 bp alleles. Moreover, the AG insertion immediately at the end of CT-repeat region was also found in 216, 200, 174, and 164 bp alleles.     

Ploidy variation in the cultivated yams (Dioscorea cayenensis-Dioscorea rotundata complex) from Cameroon as determined by flow cytometry

The ploidy levels of the twenty-two yam (Dioscorea cayenensis-D. rotundata complex) cultivars within germplasm Cameroon Guinea yam were determined by flow cytometry. Three different ploidy levels (4x,6x, 8x) were detected within the samples analysed. Fifteen cultivars were tetraploids, five were hexaploids, and two were octoploids. The cultivar group EKOTO showed a high level of ploidy variation with tetraploid, hexaploid and octoploid cultivars. The hexaploid nature of cultivars Dobnawo and Bilougnou supported the hypothesis that they are hybrids between cultivars of the EKOTO group and either the KPE or BAKOKAE groups. This revised version was published online in July 2006 with corrections to the Cover Date.