Comparative Proteomic Analysis to Reveal Key Maize Low Nitrogen Responsive Genes

Two maize hybrid cultivars contrasting in low nitrogen tolerance (low nitrogen-tolerant XY335 and low nitrogen-sensitive HN178) were used in this study . The experiment was carried out at Xinji Experimental Station (43º31′N, 124º48′E) of Hebei Agricultural University. The top 0-20 cm of the soil used contained organic matter 17.79 g·kg-1, total nitrogen 1.21 g·kg-1, alkali hydrolyzed nitrogen 64.9 mg.kg-1, available phosphorus 23.8 mg·kg-1, and available potassium 120.6 mg·kg-1. The experiment adopted a split plot design, with varieties as the main plot and nitrogen fertilizer as the sub-plot. There were 2 varieties for testing: XY335 and HN138. Two levels of nitrogen supply: N0 (0 kg N ha-1) and N240 (240 kg N ha-1), replicated three times. Each plot had 6 rows, with the row length measuring 20 m, and the row spacing of 60 cm, giving the plot area of 72 m2. The planting density was 67,500 plants ha-1. Nitrogen fertilizer used was urea (46% N), and 50% was applied before sowing and at the flared stage, respectively. During the grain filling stage, leaf tissues of three biological replicates were collected from control and treatment conditions, and immediately frozen in liquid nitrogen for subsequent proteomics analysis.