Heterosis is observed for most phenotypic traits and developmental stages in many plants. In this study, the embryos, from germinating seeds after 24 h of soaking, for five elite maize hybrids and their parents were selected to unravel the genetic basis of heterosis using 2-D proteomic method. In total, 257 (80.06%), 363 (58.74%), 351 (79.95%), 242 (54.50%), and 244 (46.30%) nonadditively expressed proteins were identified in hybrids Zhengdan 958, Nongda 108, Yuyu 22, Xundan 20, and Xundan 18, respectively. The nonadditive proteins were divided into above high-parent (++; 811, 55.66%), high-parent (+; 121, 8.30%), partial dominance (+-; 249, 17.09%), low-parent (-; 30, 2.06%), below low-parent (- -; 62, 4.26%), and D (different; 184, 12.63%) expression patterns. The observed patterns indicate the important roles of dominance, partial dominance, and overdominance in regulating seed germination in maize. Additionally, 54 different proteins were identified by mass spectrometry and classified into nine functional groups: metabolism (9), cell detoxification (8), unknown functional proteins (8), chaperones (7), signal transduction (6), development process (5), other (5), transporter (3), and stress response (3). Of these, the most interesting are those involved with germination-related hormone signal transduction and the abscisic acid and gibberellin regulation networks.
The grain filling rate is closely associated with final grain yield of maize during the period of maize grain filling. To identify the key microRNAs (miRNAs) and miRNA-dependent gene regulation networks of grain filling in maize, a deep-sequencing technique was used to research the dynamic expression patternsof miRNAs at four distinct developmental grain filling stages in Zhengdan 958, which is an elite hybrid and cultivated widely in China. The sequencing result showed that the expression amount of almost all miRNAs was changing with the development of the grain filling and formed in seven groups. After normalization, 77 conserved miRNAs and 74 novel miRNAs were co-detected in these four samples. Eighty-one out of 162 targets of the conserved miRNAs belonged to transcriptional regulation (81, 50%), followed by oxidoreductase activity (18, 11%), signal transduction (16, 10%) and development (15, 9%). The result showed that miRNA 156, 393, 396 and 397, with their respective targets, might play key roles in the grain filling rate by regulating maize growth, development and environment stress response. The result also offered novel insights into the dynamic change of miRNAs during the developing process of maize kernels and assistedin the understanding of how miRNAs are functioning about the grain filling rate.
Grain filling during the linear phase contributes most of the dry matter accumulated in the maize kernel, which in turn determines the final grain yield. Endosperms and embryos of three elite maize hybrids (Zhengdan 958, Nongda 108, and Pioneer 335) were sampled 17, 22, 25, and 28 days after pollination, during the linear phase of grain filling, for proteomic analysis to explore the regulatory factors critical for grain filling rate. In total, 39 and 43 protein spots that showed more than 2-fold changes in abundance at P<0.01 between any two sampling stages in the endosperm and embryo were analyzed by protein mass spectrometry. The changing patterns in expression index of these proteins in the endosperm were evenly distributed, whereas up-regulation patterns predominated (74%) in the embryo. Functional analysis revealed that metabolism was the largest category, represented by nine proteins in the endosperm and 12 proteins in the embryo, of the proteins that significantly changed in abundance. Glycolysis, a critical process both for glucose conversion into pyruvate and for release of free energy and reducing power, and proteins related to redox homeostasis were emphasized in the endosperm. Additionally, lipid, nitrogen, and inositol metabolism related to fatty acid biosynthesis and late embryogenesis abundant proteins were emphasized in the embryo. One protein related to cellular redox equilibrium, which showed a more than 50-fold change in abundance and was co-localized with a quantitative trait locus for grain yield on chromosome 1, was further investigated by transcriptional profile implying consistent expression pattern with protein accumulation. The present results provide a first step towards elucidation of the gene network responsible for regulation of grain filling in maize.
The influence of nitrogen (N) deficiency on tolerance mechanisms in seedlings of two maize hybrids (Xu178× Huang-C and Xu178×Zong3) and their parental inbred lines (Xu178, Huang-C and Zong3), which show different nitrogen use efficiency (NUE), was investigated using physiological measurements combined with global proteomics profiling. The root fresh weight and chlorophyll a/b ratio were reduced significantly in Huang-C (low NUE) under 0.002 mM nitrate treatment for 10 days, whereas no significant change in these two traits was observed in Xu178 (high NUE) under the same treatment compared with N-sufficient treatment. Fifty and 56 protein spots, which showed more than two-fold changes in abundance at P<0.01 under low-N treatment compared with the control in the roots and leaves, respectively, were analyzed by protein mass spectrometry. Analysis of protein expression patterns revealed that proteins associated with carbohydrate metabolism, nucleotide metabolism, amino acid metabolism, disease/ defense, and photosynthesis may be involved in N-deficiency responses. Low-N treatment led to an increased abundance of glutamine synthetase and transcripts in the root to improve the efficiency of N assimilation in the inbred line with HNE, and affected photosynthetic carbon fixation and starch metabolism in the leaves and consequently seedling growth.
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