They were Shiluan 02-1 (HMW-GS 1Ax1, 1Bx7 + 1By9, 1Dx5 + 1Dy10) and Jinan 17 (1Ax1, 1Bx7 + 1By8, 1Dx4 + 1Dy12) with strong gluten strength, Yannong 24 (1Ax1, 1Bx7 + 1By8, 1Dx5 + 1Dy10) with medium gluten strength, Lumai 21 (1Ax1, 1Bx7 + 1By8, 1Dx5 + 1Dy10)

with weak gluten strength. Shiluan 02-1, Yannong 24, and Lumai 21, were used in the growing season of 2010–2011. The 0–20 cm soil layer contained 83.6 mg kg− 1 of available nitrogen, 18.2 mg kg− 1 of available phosphate and 95.2 mg kg− 1 of available potassium. Wheat cultivars Jinan 17 and Lumai 21 were used in the 2009–2010 growing season when the soil contained available nitrogen-phosphate-potassium at 81.5, 17.6 PD0325901 concentration and 93.6 mg kg− 1, respectively. Two contrasting water regimes (irrigated and rainfed) were used. The irrigated treatment was two irrigations with the total water amount of 1500 m3 ha− 1 over the whole growth period (750 m3 ha− 1 each at jointing and booting stages, respectively), whereas the rainfed treatment had no irrigation. The moisture content in soil after anthesis is shown in Fig. 1. The experiment was a complete randomized block design with three replicates. Plot dimension was 3 m × 3 m. Plants were sown on 12 October 2010 and 15 October 2009, respectively, at a density of 180 seeds m− 2. Normal crop farming practices were implemented to minimize pest, disease and weed incidence.

CH5424802 After full heading, spikes flowering on the same date were labeled with thread. At maturity (14 June 2011 and 15 June 2010, respectively), the labeled heads were sampled and used to determine the GMP particle distributions. GMP and HMW-GS contents were also determined. The content Wilson disease protein of GMP was analyzed as follows: 0.05 g of flour was dispersed into and mixed with 1 mL of SDS and then centrifuged at 15,500 ×g for 15 min using an Allegra X-64R centrifuge (Beckman, San Francisco, CA, USA) and the supernatant was retained. Glutenin macropolymer content was measured using TU-1901

dual-wavelength spectrophotometer (Persee Instruments, Beijing, China). Glutenin macropolymer content was calculated using a set of Kjeldahl protein values. Glutenin macropolymer-gel was isolated by dispersing 1.4 g of defatted flour in 0.05 mol L− 1 SDS (pasteurized, 28 mL) and then centrifuged at 80,000 ×g for 30 min at 20 °C using a Beckman L-60 ultracentrifuge (Beckman, San Francisco, CA, USA) as described [16]. The GMP gel-layer was collected from the top of the supernatant. For Coulter laser particle size analysis, 1 g of GMP-gel was added to 8 mL of 0.05 mol L− 1 SDS solvent. The tube was sealed and placed on a roller-bank for 3 h at room temperature and analyzed with a Coulter Laser LS13320 (Beckman Coulter Instruments, San Francisco, CA, USA). The GMP surface area distribution and volume distribution were measured and calculated from the resulting pattern. Quantification of HMW-GS was performed according to the following method [17].