8�C8.0. The seeds were sown in 2m2 plots with a row space of 20cm. The sowing dates were early sown (ES) (15 October), timely sown (TS) (15 November), and late sown (LS) (15 December) in 2007 (year I) and 2008 (year II). The experimental design was a randomized complete block with three replications. Under the irrigated treatment, plants were watered throughout the period selleck inhibitor from sowing to maturity according to the recommended agronomic practices [12]. All irrigations were withheld from the plants subjected to the drought treatment except the presowing irrigation for field preparation. Therefore, the drought-treated plants received water only available through rainfall. The weather data of total rainfall and evapotranspiration rate for the crop season in both years was collected from the field meteorological observatory.
Total rainfall as an average of two years received by the crop from sowing to maturity was 94, 81, and 82mm, while during grain filling period was 23, 55, and 45mm for early, timely and late sown crops, respectively. During the crop season rainfall was scanty while evapotranspiration was higher, which helped in the development of severe drought stress.2.2. Grain PlumpnessWheat grains were separated according to the grain diameters (>2.8mm and <2.5mm) using grain sorter (Sortimat, Germany) fitted with respective sized meshes. The grains of different diameters were collected and then weighed on an electronic balance and expressed in percentage.2.3. Test Weight Test weight was determined using the apparatus developed by the Directorate of Wheat Research, Karnal, India, which employs a standard container of 100mL capacity.
The container is filled with the sample of wheat grains by removing all shrunken and broken kernels and other foreign material. The grains were weighed and the Drug_discovery test weight expressed in kg/hectoliter (hl) [13].2.4. Milling and Protein FractionationGrains from each treatment were finally milled using Cyclotec 1093 sample mill (Foss, Tecator, Sweden) to obtain wheat whole meal in both the years. Protein fractionation was carried out from wheat whole meal according to Triboi et al. [10] with a little modification. Briefly, the protein fractions albumin-globulin, amphiphil, gliadin, and glutenin were sequentially extracted from 800mg of flour. During each extraction step, the samples were continuously shaken in a volumetric flask (100mL) placed in a temperature controlled shaker (orbital shaking incubator CIS-24, Remi, India) for 60min. Soluble and insoluble fractions were separated by centrifugation at 8,000 �� g (Sigma, USA) for 30min at the extraction temperature. Albumins-globulins were extracted at 4��C with 25mL 0.05M NaCl, 0.05M sodium phosphate buffer pH 7.8.