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Cereal Research Communications
Authors: J. Bányai, P. Szűcs, I. Karsai, K. Mészáros, Cs. Kuti, L. Láng, and Z. Bedő

A total of 96 winter wheat ( Triticum aestivum L.) cultivars registered in Hungary were analysed using 15 wheat microsatellite markers located on different chromosome arms. Analyses revealed 91 SSR alleles with sizes ranging from 123–239 base pairs. The total number of alleles per locus ranged from 2 (Gwm664 and Gwm415) to 11 (Gwm219) with an average number of 6.1. The polymorphic information content (PIC) values ranged from 0.06 to 0.85 with an average number of 0.60 for all markers. Several markers included allele sizes characteristic of a single or a small number of cultivars. At most 9 SSR markers were required to distinguish the 96 cultivars, so the simple sequence repeats could serve as a relatively cheap, rapid method for identifying winter wheat cultivars.

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A quick and reproducible tool for cultivar identification is useful to assess in certified seed production programs and to resolve legal conflicts. Simple Sequence Repeats (SSRs) have been the elected markers to carry out cultivar identification studies. The main aim of this research was to define the minimum number of SSR markers to distinguish 80 durum wheat cultivars. Preliminary, an analysis of 11 SSRs informativeness was carried out on a subset of 28 durum wheat cultivars. The discriminating ability of each primer was estimated both with Polymorphism Information Content (PIC) and with Resolving power (Rp). Rp resulted the best parameter for assessing the discriminatory power of SSR primers (r=0.94***; P≤0.001). The marker Xwmc597 was able to discriminate all the 28 cultivars. Successively, 80 genotypes were analysed using three SSR markers with the higher Rp value. Two SSRs were able to distinguish all the 80 genotypes. Particularly, Xwmc597 was able to distinguish 69/80 genotypes while Xwmc415 identified the other cultivars. An identification key was obtained combining the data of these two markers.

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During the last century wheat landraces were replaced by modern wheat cultivars leading to a gradual process of genetic erosion. Landraces genotyping and phenotyping are strategically useful, as they could broaden the genetic base of modern cultivars. In this research, we explored Single Nucleotide Polymorphism (SNP) markers diversity in a collection of common and durum wheats, including both landraces and Italian elite cultivars. A panel of 6,872 SNP markers was used to analyze the genetic variability among the accessions, using both the Principal Components Analysis (PCA) and the Neighbour Joining clustering method. PCA analysis separated common wheat accessions from durum ones, and allowed to group separately durum landraces from durum elite cultivars. The Neighbour joining clustering validated PCA results, and moreover, separated common wheat landraces from common elite cultivars. The clustering results demonstrated that Italian durum landraces were poorly exploited in modern breeding programs. Combining cluster results with heterozygosity levels observed, it was possible to clarify synonymy and homonymy cases identified for Bianchetta, Risciola, Saragolla, Timilia and Dauno III accessions. The SNP panel was also used to detect the minimum number of markers to discriminate the studied accessions. A set of 33 SNPs were found to be highly informative and used for a molecular barcode, which could be useful for cultivar identification and for the traceability of wheat end-products.

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DNA methylation polymorphism among nine elite maize inbred lines was assessed by inter-simple sequence repeat (ISSR) fingerprinting on intact DNA and DNA digested by either of a pair of methylation-sensitive isoschizomers, Hpa II and Msp I. It was found that, along with distinct genetic differentiation, extensive DNA methylation polymorphism exists among the nine inbred lines. The line-specific methylation patterns are homogeneous within each line, indicating their usefulness as molecular markers for cultivar identification. DNA sequences underlying the DNA methylation variations include a high proportion of coding genes. Cluster analysis, however, indicates the existence of incongruence between DNA methylation polymorphism and known-pedigree of the maize inbred lines.

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Acta Alimentaria
Authors: G. Balázs, I. Baracskai, M. Nádosi, A. Harasztos, F. Békés, and S. Tömösközi

The utilisation of microchip capillary electrophoresis has the potential to improve the capability of high throughput sample analysis of biomolecules. The aim of this study was to review this capability for cereal protein analysis.The commercially available lab-on-a-chip (LOC) technology was characterised in the separation of total proteins extracted from whole wheat meals. Important analytical parameters (such as repeatability) of both qualitative (molecular size estimation) and quantitative (relative percentage of total protein) aspects of LOC data were determined and discussed in the light of the need of possible applications. It revealed that the LOC has very good repeatability and reproducibility parameters; however the non-globular structure of the proteins can highly affect the sizing accuracy. Among other applications, the profiles were found to be suitable for wheat cultivar identification and to monitor environment related alterations on protein composition.After a confirmation process the LOC can be an appropriate tool for fast protein profile screening in cereal science and technology in diverse applications, and it can complement the conventional methods of analysis.

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Seed protein pattern of control and M 10 mutant soybean ( Glycine max [L.] Merr.) lines in defatted and non-defatted raw flour was studied after 60% 2-propanol extraction, SDS-PAGE separation, colloidal staining and densitometric evaluation to detect a new variant of the protein KTI and/or BBI, furthermore to find new protein(s) of low molecular weight. Electrophoretic separation of defatted and non-defatted control soybean samples showed the same protein patterns. On the densitograms of mutant lines quantitative and qualitative differences could be observed. Defatted raw soy samples reflected more differences in the number of peaks than non-defatted ones. Beside soy trypsin inhibitors, several more soy proteins of low molecular weight are dissolved. KA mutant line 9 has a unique 2-propanol soluble protein pattern, and a new protein band of Rf=0.37 compared to the control line. Sixty percent 2-propanol soluble soybean seed proteins are suitable for cultivar identification and characterization, furthermore to distinguish soybean lines of the same origin.

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14 Bushuk W and R R Zillman, 1978: Wheat cultivar identification by gliadin electrophoregrams. I Apparatus, method and nomenclature. Can. J. Plant Sci. 58 : 505

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De Villiers, O.T., Bosman, M. 1993. Wheat cultivar identification by electrophoretic analysis of gliandin protein. S. Afr. J. Plant Soil 10 :99–104. Bosman M. Wheat cultivar

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. 2007 87 763 777 Zillman, R.R., Bushuk, W. 1979a. Wheat cultivar identification by gliadin electrophoregrams. II. Effects

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. Barley cultivar identification by electrophoretic analysis of hordein proteins. II. Catalogue of electrophoregram formulae for Canadian-grown barley cultivars. Can. J. Plant Sci. 60: 859–870. Laberge D

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