Identification, expression and DNA variation analysis of high affinity nitrate transporter NRT2/3 gene family in Sorghum bicolor / 生物工程学报

Nitrate is the main form of inorganic nitrogen that crop absorbs, and nitrate transporter 2 (NRT2) is a high affinity transporter using nitrate as a specific substrate. When the available nitrate is limited, the high affinity transport systems are activated and play an important role in the process of nitrate absorption and transport. Most NRT2 cannot transport nitrates alone and require the assistance of a helper protein belonging to nitrate assimilation related family (NAR2) to complete the absorption or transport of nitrates. Crop nitrogen utilization efficiency is affected by environmental conditions, and there are differences between varieties, so it is of great significance to develop varieties with high nitrogen utilization efficiency. Sorghum bicolor has high stress tolerance and is more efficient in soil nitrogen uptake and utilization. The S. bicolor genome database was scanned to systematically analyze the gene structure, chromosomal localization, physicochemical properties, secondary structure and transmembrane domain, signal peptide and subcellular localization, promoter region cis-acting elements, phylogenetic evolution, single nucleotide polymorphism (SNP) recognition and annotation, and selection pressure of the gene family members. Through bioinformatics analysis, 5 NRT2 gene members (designated as SbNRT2-1a, SbNRT2-1b, SbNRT2-2, SbNRT2-3, and SbNRT2-4) and 2 NAR2 gene members (designated as SbNRT3-1 and SbNRT3-2) were identified, the number of which was less than that of foxtail millet. SbNRT2/3 were distributed on 3 chromosomes, and could be divided into four subfamilies. The genetic structure of the same subfamilies was highly similar. The average value of SbNRT2/3 hydrophilicity was positive, indicating that they were all hydrophobic proteins, whereas α-helix and random coil accounted for more than 70% of the total secondary structure. Subcellular localization occurred on plasma membrane, where SbNRT2 proteins did not contain signal peptides, but SbNRT3 proteins contained signal peptides. Further analysis revealed that the number of transmembrane domains of the SbNRT2s family members was greater than 10, while that of the SbNRT3s were 2. There was a close collinearity between NRT2/3s of S. bicolor and Zea mays. Protein domains analysis showed the presence of MFS_1 and NAR2 protein domains, which supported executing high affinity nitrate transport. Phylogenetic tree analysis showed that SbNRT2/3 were more closely related to those of Z. mays and Setaria italic. Analysis of gene promoter cis-acting elements indicated that the promoter region of SbNRT2/3 had several plant hormones and stress response elements, which might respond to growth and environmental cues. Gene expression heat map showed that SbNRT2-3 and SbNRT3-1 were induced by nitrate in the root and stem, respectively, and SbNRT2-4 and SbNRT2-3 were induced by low nitrogen in the root and stem. Non-synonymous SNP variants were found in SbNRT2-4 and SbNRT2-1a. Selection pressure analysis showed that the SbNRT2/3 were subject to purification and selection during evolution. The expression of SbNRT2/3 gene and the effect of aphid infection were consistent with the expression analysis results of genes in different tissues, and SbNRT2-1b and SbNRT3-1 were significantly expressed in the roots of aphid lines 5-27sug, and the expression levels of SbNRT2-3, SbNRT2-4 and SbNRT3-2 were significantly reduced in sorghum aphid infested leaves. Overall, genome-wide identification, expression and DNA variation analysis of NRT2/3 gene family of Sorghum bicolor provided a basis for elucidating the high efficiency of sorghum in nitrogen utilization.

Divergence of flowering genes in soybean.

Soybean genome sequences were blasted with Arabidopsis thaliana regulatory genes involved in photoperioddependent flowering. This approach enabled the identification of 118 genes involved in the flowering pathway. Two genome sequences of cultivated (Williams 82) and wild (IT182932) soybeans were employed to survey functional DNA variations in the flowering-related homologs. Forty genes exhibiting nonsynonymous substitutions between G. max and G. soja were catalogued. In addition, 22 genes were found to co-localize with QTLs for six traits including flowering time, first flower, pod maturity, beginning of pod, reproductive period, and seed filling period. Among the genes overlapping the QTL regions, two LHY/CCA1 genes, GI and SFR6 contained amino acid changes. The recently duplicated sequence regions of the soybean genome were used as additional criteria for the speculation of the putative function of the homologs. Two duplicated regions showed redundancy of both flowering-related genes and QTLs. ID 12398025, which contains the homeologous regions between chr 7 and chr 16, was redundant for the LHY/CCA1 and SPA1 homologs and the QTLs. Retaining of the CRY1 gene and the pod maturity QTLs were observed in the duplicated region of ID 23546507 on chr 4 and chr 6. Functional DNA variation of the LHY/CCA1 gene (Glyma07g05410) was present in a counterpart of the duplicated region on chr 7, while the gene (Glyma16g01980) present in the other portion of the duplicated region on chr 16 did not show a functional sequence change. The gene list catalogued in this study provides primary insight for understanding the regulation of flowering time and maturity in soybean.

DNA Variation of Helicobacter Pylori in the Gastroduodenal Disease / 대한내과학회지

BACKGROUND: The evidence for H. pylori as a gastrointestnal pathogen is now very strong, if not overwhelming. Among the pathogenic factors of H. pylori, flagella and urease are considered to be major factors causing the gastrododenal disease. We observed the gene diversity of H. pylori using the PCR-amplified 1.4Kb fla A gene and 0.9Kb ure B gene and examined the relationship between the gene pattern and the gastroduodenal disease. METHOD: Fifty-one cases of isolated strains were cultured at the Helicobacter-selective blood agar plates. To compare the gene diversity among the isolates of gastroduodenal disease genotypes was analyzed by PCR-based RFLP. 1.4Kb fla A gene and 0.9Kb ure B genes from isolates were amplified by PCR and digested with Hae 3 restriction enzymes to observe the restriction fragment length polymophysm. Protein patterns were also compared to examine the antigenic variations. Total cell proteins, and octyl-glucose extracts from isolates were analyzed by SDS-PAGE gel electrophoresis. RESULTS: 41 cases (80.4%) of H. pylori were isolated in the 51 cases of gastroduodenal diseases. We could classify theses isolates 3 types of PCR-RFLP in the fla A gene, 900+500bp, 500+500+400bp, 600+800bp, and 9 types in the ure B gene. PCR-RFLP in the fla A gene and ure B gene of the isolates was different from the standard strain of Australia and the genetic diversity was not related to the types of the gastroduodenal disease. We demonstrated variations in the protein pattern and antigenic profiles among the isolates by SDS-PAGE analysis. These data also did not show any relationship between protein pattern and types of gastroduodenal diseases. CONCLUSION: Tese studies showed many different gene diversity in the flagella and urease gene without any relationship with the types of gastoduodenal disease. And variable protein pattern were noted among the strains of H. pylori. Further studies to demonstrate the pathgenecity of H. pylori should be continued even if there was no relationship between the genomic diversity of the flagella or urease and the types of gastroduodenal disease.