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Pakistan Journal of Biological Sciences
Year: 2004  |  Volume: 7  |  Issue: 6  |  Page No.: 984 - 994

The Roles of Seed Proteins and RAPD-PCR in Genotyping Variabilities of Some Wheat (Triticum vulgare L.) Cultivars

Maher M. Shehata    

Abstract: Traditional identification of wheat cultivars relies on the assessment of agronomic traits of the adult plant. This leads to a significant delay of time, constraints to breeders in the surveillance of germplasm and a risk for growers and exporters. Seed storage protein and DNA fingerprinting diversity as revealed by variation in SDS-PAGE and RAPD-PCR, respectively has been used to reassess the genotypic differences, relationships and discriminating between eight wheat (Triticum vulgare L.) cultivars (Banisewef 3, Gemmiza 7, Giza 164 and 168, Sakha 8, 69 and 93 and Sids 1) grown in Egypt. A total number of 46 protein bands, with molecular weight ranging between 160.52 and 3.86 kDa were recorded in the electrophenograms of the cultivars studied. SDS-PAGE profiles showed slight differences at high and low molecular weights and high differences at mid molecular weights. The highest number of bands (30) was recorded in Banisewef 3 and the lowest number (21) was recorded in Sids 1. Eight common bands (M.wts.; 155.32, 139.50, 76.35, 29.25, 19.25, 8.20, 7.20 and 6.38 kDa) were recorded in all samples. Six unique bands were recorded in cultivars Banisewef 3 (M.wts: 132.69 and 73.50 kDa), Sakha 8 (M.wt.: 38.50 kDa) and Sakha 69 (M.wts.: 88.59, 63.20 and 24.85 kDa). In the RAPD-PCR analysis twenty five random decamer (10 mer) primers were screened, but only eight primers (OPB-01, -02, -03, -08 and OPI-03, -09, -14 and -18) were able to generate distinguishable, reproducible and repeatable informative products among the DNA samples of the studied genotypes. A total of 67 polymorphic bands out of 86 ones were generated by the eight primers. Nearly every cultivar has a different DNA fingerprint. Each cultivar has one or more novel sequences not found in other cultivars. These bands may be used as genetic markers for identification of these cultivars using the same primer. A total of 8 polymorphic bands were scored as unique ones. Primers OPI-09 and -03 were found to be the most effective in generating unique bands. The former primer generated 4 unique bands in Banisewef and Giza 164 cultivars while the latter primer produced 2 unique bands in Giza 169 and Sakha 93 cultivars. Increasing and changing the number of cycles in PCR from 30 to 40, improve our results. These bands were used as binary characters and analyzed by the NTSYS-pc. Program Package using the UPGMA clustering method. This analysis indicates that, the eight cultivars are distinct. SDS-PAGE and RAPD-PCR data were combined together and used to construct a dendrogram that estimates the relationship and the relative genetic similarities among the studied wheat cultivars. Based on this dendrogram, the studied cultivars were separated into two main groups. The first group comprises three cultivars, while the second includes five cultivars. The first group was further subdivided into two subgroups; the first subgroup comprises cultivar Sakha 8, whereas the second subgroup includes cultivars Sakha 69 and Sakha 93. The second group was further subdivided into two subgroups; the first subgroup comprises cultivars Giza 169 and Giza 164, whereas the second subgroup includes cultivars Sids 1, Gemmiza 7 and Banisewef 3. Combination of all data provides a considerable potentiality for discriminating each wheat cultivar by one or more unique bands or a group of combined class pattern. As a result of this investigation, we may expect that the SDS-PAGE, RAPD and the subsequent banding patterns computed using appropriate programs, would be useful for the establishment of phylogenetic relationships among a set of Egyptian wheat cultivars. These tools could be used as complementary of traditional methods of identification of phenotypic traits for the control of registered cultivars in the trade market. This will help in collection and cataloguing of the germplasm in the form of a germplasm bank.

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