Protein quantification and enzyme activity estimation of Pakistani wheat landraces

Abstract

Wheat is a major food grain in Pakistan having a prominent role in agriculture as well as the economic status of the country. In the current study, seeds of 99 wheat landraces were characterized for the quantification of seed storage proteins (Albumins, Globulin, Gliadins, and Glutenin), enzyme activities of antioxidant enzymes i.e. Ascorbate peroxidase (APX), Catalase (CAT), Superoxide dismutase (SOD), Peroxidase (POD), one hydrolytic enzyme Protease (PROT) and non-enzymatic antioxidant enzyme Ascorbic acid (AsA). The landraces were categorized into low, medium, and high based on protein concentration and enzymes activities/content. The majority of the landraces were placed in the medium category. However, for the AsA parameter majority of the landraces were placed in the low category. The highest concentration of total extracted protein (184.88±0.7 mg/g. wt.), globulins (21.35 ±0.43 mg/g. wt.) and glutenin (20±0.04 mg/g. wt.) as well as the high activity of SOD (303 ±16.80 Units/g. wt.), and Ascorbic acid (533±36.1 Units/g. wt.) was identified in the wheat landrace "11757" collected from district Panjgur (Balochistan). The wheat landrace "11760", collected from district Kech (Balochistan), contained the highest albumins concentration (65.42±0.02 mg/g. wt.) and highest activity for CAT (589.5±61.20 Units/g. wt.). The highest activity of POD (32341± 91.3) and PROT was observed in seeds of the wheat landrace "11618" collected from the Gilgit Baltistan region of Pakistan. The principal component analysis showed that the great variations existed for the tested parameters among the wheat landraces. The landraces with a high concentration of seed storage proteins and antioxidant enzyme activities can be used for breeding purposes to improve the nutrimental quality of wheat cultivars.

Mining the Wheat Grain Proteome

“…The powder:volume ratio that produced the best results during our tests was 25 µL/mg, same ratio used by [ 19 , 20 ]. Usage of more concentrated ratios are often reported in the literature: 4 µL/mg (50 mg: 200 µL) [ 13 ]; 5 µL/mg (100 mg:0.5 mL) [ 21 ]; 6.7 µL/mg (30 mg:200 µL) [ 8 ]; 10 µL/mg [ 10 , 11 , 15 , 18 , 22 , 23 , 24 ]; 16 µL/mg (50 mg:800 µL or 100 mg:1.6 mL) [ 14 , 25 ]; and 20 µL/mg (0.5 g:10 mL) [ 26 ], although less frequently, more diluted ratios are mentioned: 33 µL/mg (30 mg:1 mL) [ 27 ]; 40 µL/mg; (100 mg:4 mL) [ 28 ]; 50 µL/mg (300 mg:15 mL) [ 29 ]; and 100 µL/mg (10 mg:1 mL) [ 30 ]. Our optimal ratio fitted within the 4 to 100 µL/mg range reported in published literature.…”

“…Both buffers yielded comparable identification rates, with a slight advantage to urea (8846 peptides; 1483 accessions) relative to Tris-HCl (8632 peptides; 1405 accessions); 79% of the accessions were shared across both buffers [ 23 ]. Other reports have employed the sequential extraction method devised by Osborne in 1924 [ 31 ] to recovery successive protein fractions using first an aqueous buffer to extract albumins, second a salt solution to retrieve globulins, third an ethanol/water mixture to solubilise gliadins, and finally a propanol solution to extract glutenins [ 8 , 10 , 12 , 15 , 19 , 20 , 21 , 22 , 24 , 26 , 27 , 29 , 30 , 32 ]. Another multistep procedure involved an initial phenol/ammonium acetate phase partition followed by solvent precipitation and Tris-HCl buffer resuspension [ 7 , 8 , 18 , 33 ].…”

Assessment of Four Portuguese Wheat Landrace Diversity to Cope With Global Warming

“…It is relevant that although these landraces were not submitted to breeding programs, their values of protein content are very acceptable and similar to the ones reported for commercial varieties. Recently, the screening of Pakistani wheat landraces also found several traditional genotypes with high storage protein concentration, pointing out their potential to improve the nutritional quality of modern wheat commercial genotypes ( Mughal et al, 2020 ).…”

Coupled online sequential extreme learning machine model with ant colony optimization algorithm for wheat yield prediction

“…The current wheat yield prediction and forecasting methods adopted by the Pakistan Government have been reported to be highly inaccurate 12 . In 2005, poor wheat yield predictions of Pakistan where the actual production was relatively small equated to the estimated yield, resulted in an inflated local market rate and prompted higher importation of wheat 13 , 14 .…”