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Molecular and Biochemical Characterization of Bangladeshi Wheat Varieties for Bread Making Quality (2014-2015)

Sbatie Lama
Biotechnology Division, BARI, Joydebpur, Gazipur

M.A.Y. Akhond
Biotechnology Division, BARI, Joydebpur, Gazipur

Abstract/ Executive Summary:

The experiment was conducted with a view to characterizing 26 wheat varieties for bread-making quality through Polymerase Chain Reaction (PCR) method using gene-specific markers for 7 High Molecular Weight Glutenin Subunit (HMW-GS). These varieties were previously characterized through Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) method. Ax2*, Ax1, Dx5, Bx7, BxFp, ZSBy8F5/R5, ZSBy9Af1/R3 gene-specific markers were used to identify Glu-Ax2*, Glu-Ax1, Glu-Dx5, Glu-Bx7, Glu-Bx17, Glu-By8 and Glu-By9 alleles respectively. For A1 locus 22 varieties were found containing the Glu-Ax2* allele whereas, the rest of the four varieties containing the Glu-Ax1 allele. In Glu-D1 locus, 16 varieties were identified containing the Dx5+Dy10 allele. For the Glu-B1 locus, 5 varieties were found with the Bx17 allele. Bx7 allele is usually associated with either of two subunits, By8 or By9. Twenty-two varieties were found with the Bx7 allele however no PCR amplifications were found for By8 and By9.

Keywords: DNA extraction, Genomic DNA, Agarose gel, Wheat

Location:

Start Date: 00-00-2014

End Date: 00-00-2015

Program Area: Variety and Species

Commodity: Wheat

Objectives:

The study was aimed to identify HMW-GS of wheat varieties grown in Bangladesh in relation to the bread-making quality using PCR-based techniques.

Methodology:

The 26 genotypes were established at the greenhouse. Young-tender leaves with 10 days old were harvested for DNA extraction. Genomic DNA was isolated by following the protocol of the Genomic DNA Mini Kit. Polymerase chain reaction (PCR) was carried out in a 20 μl reaction volume. The PCR products were separated by electrophoresis in 1% agarose gel and visualized over ultraviolet light in a gel documentation system.

Source of Information: Annual Research Report 2014-2015, Biotechnology Division, BARI, Joydebpur, Gazipur

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The 26 wheat varieties were tested to ensure the presence of specific HMW-GS using gene-specific primers for Ax1, Ax2*, Dx5, Bx17+By18, Bx7, By8 and By9. Ax gene: The PCR produced one sharp band for Ax2* genotypes in 22 varieties with no PCR amplification for Ax1 genotypes. Specific primer for Ax1 produced a similar size product as Ax2* for the presence of Ax1 genotype in 5 varieties. Dx gene: According to Liu et al. (2008) the Glu-D1locus has the largest effect on bread-making quality. Using the Dx5 primers, a sharp DNA band (478 bp) was amplified only in the presence of the Dx5 gene in 16 varieties (Fig. 2). Bx gene: PCR primers specific for the By17 genotype produced one strong DNA band for varieties containing the Bx17 gene (product size 669 bp) in 5 varieties (Fig. 3) and two bands for non-Bx17 genes (630 and 766 bp). Bx17 allele is always present in the combination with the By18 allele. The presence of allelic pair Bx17+1By18 in wheat varieties is good for bread-making quality (Xu. et. al., 2008).

Knowledge Management: Report/Proceedings