Cloning and characterization of maize ZmSPK1, a homologue to nonfermenting1-related protein kinase2

SnRK2s play important roles in plant stresses responses. One full-length cDNA encoding a SnRK2b homologue was isolated from maize by RT-PCR and named as ZmSPK1 (for stress-induced protein kinase). The ZmSPK1 protein has 364 amino acids with an estimated molecular mass of 41.8 KD and an isoelectric point of 5.8. The deduced protein sequence has the closest identities to the members of SnRK2b group. RT-PCR analysis showed that the ZmSPK1 expression was induced by mannitol, salt and abscisic acid (ABA). Furthermore, in different tissues the ZmSPK1 showed different expression patterns and was most abundant in reproductive organs. These results suggested that ZmSPK1 might play multiple roles in abiotic stress resistance pathways, as well as in plant reproductive development.Key words: Zea mays L., SnRK2b, expression pattern, abiotic stres

Development of Omni InDel and supporting database for maize

Insertions–deletions (InDels) are the second most abundant molecular marker in the genome and have been widely used in molecular biology research along with simple sequence repeats (SSR) and single-nucleotide polymorphisms (SNP). However, InDel variant mining and marker development usually focuses on a single type of dimorphic InDel, which does not reflect the overall InDel diversity across the genome. Here, we developed Omni InDels for maize, soybean, and rice based on sequencing data and genome assembly that included InDel variants with base lengths from 1 bp to several Mb, and we conducted a detailed classification of Omni InDels. Moreover, we screened a set of InDels that are easily detected and typed (Perfect InDels) from the Omni InDels, verified the site authenticity using 3,587 germplasm resources from 11 groups, and analyzed the germplasm resources. Furthermore, we developed a Multi-InDel set based on the Omni InDels; each Multi-InDel contains multiple InDels, which greatly increases site polymorphism, they can be detected in multiple platforms such as fluorescent capillary electrophoresis and sequencing. Finally, we developed an online database website to make Omni InDels easy to use and share and developed a visual browsing function called “Variant viewer” for all Omni InDel sites to better display the variant distribution

Full Length Research - Cloning and characterization of maize ZmSPK1, a homologue to nonfermenting1-related protein kinase2

SnRK2s play important roles in plant stresses responses. One full-length cDNA encoding a SnRK2b homologue was isolated from maize by RT-PCR and named as ZmSPK1 (for stress-induced protein kinase). The ZmSPK1 protein has 364 amino acids with an estimated molecular mass of 41.8 KD and an isoelectric point of 5.8. The deduced protein sequence has the closest identities to the members of SnRK2b group. RT-PCR analysis showed that the ZmSPK1 expression was induced by mannitol, salt and abscisic acid (ABA). Furthermore, in different tissues the ZmSPK1 showed different expression patterns and was most abundant in reproductive organs. These results suggested that ZmSPK1 might play multiple roles in abiotic stress resistance pathways, as well as in plant reproductive development

Genome‐wide association analysis of chilling‐tolerant germination in a new maize association mapping panel

Abstract Maize is a crop that is highly susceptible to the negative effects of low temperature. Low temperature can delay seed germination and cause a decrease in seed vigor, which seriously affects seedling emergence and yield. In this study, 190 maize accessions (inbred lines) with strong germination potential at normal temperature (25°C) were selected from more than 500 accessions to construct a new association mapping panel to further investigate germination under chilling stress (5°C). We re‐sequenced the genomes of the 190 diverse accessions and obtained 4,886,919 high‐quality SNPs. We then used this data to analyze population structure, perform principal components analysis, and construct a phylogenetic tree of the new maize panel. The relative germination rate (RGR) and relative germination index (RGI) are two traits that are significantly related to chilling‐tolerant germination. Genome‐wide association analysis showed that RGR and RGI shared a major QTL, and they also shared the top SNP. There were a total of 26 significant SNPs in common. These SNPs hit directly or indirectly within 37 candidate genes. Among these 37 gene candidates, eight are homologs of genes previously reported to be related to both germination and low‐temperature stress, and another 12 genes related to low‐temperature stress or other abiotic stresses such as drought, salinity, oxidative, and high light stress. In addition, RGR and RGI had another 15 and 26 significant SNPs, respectively, which were associated with 17 and 92 candidate genes, respectively. Further qRT‐PCR analysis using 26 chilling‐tolerant and 22 chilling‐sensitive accessions implied that Zm00001eb272370, Zm00001eb272390, and Zm00001eb272400 associated with the top SNP, may play different roles during cold‐germination. Thus, our study not only established a new association mapping panel suitable for investigation of germination at low temperature but also provided valuable genetic resources for future studies to improve chilling‐tolerant maize varieties

Comparative Proteomics of Contrasting Maize Genotypes Provides Insights into Salt-Stress Tolerance Mechanisms

Salt stress is a major abiotic factor limiting maize yield. To characterize the mechanism underlying maize salt tolerance, we compared the seedling root proteomes of salt-tolerant Jing724 and salt-sensitive D9H. The germination rate and growth parameter values (weight and length) were higher for Jing724 than for D9H under saline conditions. Using an iTRAQ-based method, we identified 513 differentially regulated proteins (DRPs), with 83 and 386 DRPs specific to Jing724 and D9H, respectively. In salt-stressed Jing724, the DRPs were primarily associated with the pentose phosphate pathway, glutathione metabolism, and nitrogen metabolism. Key DRPs, such as glucose-6-phosphate 1-dehydrogenase, NADPH-producing dehydrogenase, glutamate synthase, and glutamine synthetase, were identified based on pathway enrichment and protein–protein interaction analyses. Moreover, salt-responsive proteins in Jing724 seedlings were implicated in energy management, maintenance of redox homeostasis, detoxification of ammonia, regulation of osmotic homeostasis, stress defense and adaptation, biotic cross-tolerance, and regulation of gene expression. Quantitative analyses of superoxide dismutase activity, malondialdehyde content, relative electrolyte leakage, and proline content were consistent with the predicted changes based on DRP functions. Furthermore, changes in the abundance of eight representative DRPs were correlated with the corresponding mRNA levels. Our results may be useful for elucidating the molecular networks mediating salt tolerance

Relative gene expression of the 8 MAPKKK genes in various inbred lines based on qRT-PCR analysis.

<p>To determine whether the relative expression levels of the drought stress-responsive MAPKKK genes differed among varieties induced by drought, ZD619 and the 6 inbred maize lines J24, J853, X178, E28, C8605-2, 200B, Q319 and B73 were used. Lines X178, J24 are drought-resistant lines; 200B and E28 have poor drought tolerance. The eight MAPKKK genes are <i>GRMZM2G305066</i> (<i>ZmMAPKKK18</i>), <i>GRMZM2G165099</i> (<i>ZmMAPKKK19</i>), <i>GRMZM2G476477</i> (<i>ZmMAPKKK20</i>), <i>GRMZM2G173965</i> (<i>ZmMAPKKK21</i>), <i>GRMZM2G041774</i> (<i>ZmMAPKKK22</i>), <i>GRMZM2G021416</i> (<i>ZmMAPKKK26</i>), <i>GRMZM2G063069</i> (<i>ZmMAPKKK56</i>), <i>GRMZM2G474546</i> (<i>ZmMAPKKK73</i>). * indicates significant differences in comparison with the control at P < 0.05 respectively. Error bars indicate standard deviation for three replicates.</p