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Wang R.K.,Jiujiang UniversityJiangxi Province | Zhan S.F.,Jiujiang UniversityJiangxi Province | Zhao T.J.,Nanjing Agricultural UniversityJiangsu Province | Zhou X.L.,Jiujiang UniversityJiangxi Province | Wang C.E.,Jiujiang UniversityJiangxi Province
Genetics and Molecular Research | Year: 2015

Isoflavonoids and the related synthesis enzyme, chalcone isomerase 1 (CHI1), are unique in the Leguminosae, with diverse biological functions. Among the Leguminosae, the soybean is an important oil, protein crop, and model plant. In this study, we aimed to detect the generation pattern of Leguminosae CHI1. Genome-wide sequence analysis of CHI in 3 Leguminosae and 3 other closely related model plants was performed; the expression levels of soybean chalcone isomerases were also analyzed. By comparing positively selected sites and their protein structures, we retrieved the evolution patterns for Leguminosae CHI1. A total of 28 CHI and 7 FAP3 (CHI4) genes were identified and separated into 4 clades: CHI1, CHI2, CHI3, and FAP3. Soybean genes belonging to the same chalcone isomerase subfamily had similar expression patterns. CHI1, the unique chalcone isomerase subfamily in Leguminosae, showed signs of significant positive selection as well as special expression characteristics, indicating an accelerated evolution throughout its divergence. Eight sites were identified as undergoing positive selection with high confidence. When mapped onto the tertiary structure of CHI1, these 8 sites were observed surrounding the enzyme substrate only; some of them connected to the catalytic core of CHI. Thus, we inferred that the generation of Leguminosae CHI1 is dependent on the positively selected amino acids surrounding its catalytic substrate. In other words, the evolution of CHI1 was driven by specific selection or processing conditions within the substrate. © FUNPEC-RP. Source

Yu M.,Nanjing Agricultural UniversityJiangsu Province | Xu Y.,Nanjing Agricultural UniversityJiangsu Province | Yu D.,Nanjing Agricultural UniversityJiangsu Province | Du W.,Nanjing Agricultural UniversityJiangsu Province
Journal of Reproduction and Development | Year: 2015

Many genes participate in the process of ovarian germ cell development, while the combined action mechanisms of these molecular regulators still need clarification. The present study was focused on determination of differentially expressed genes and gene functions at four critical time points in chicken ovarian development. Comparative transcriptional profiling of ovaries from embryonic day 5.5 (E5.5), E12.5, E15.5 and E18.5 was performed using an Affymetrix GeneChip chicken genome microarray. Differential expression patterns for genes specifically depleted and enriched in each stage were identified. The results showed that most of the up- and downregulated genes were involved in the metabolism of retinoic acid (RA) and synthesis of hormones. Among them, a higher number of up- and downregulated genes in the E15.5 ovary were identified as being involved in steroid biosynthesis and retinol metabolism, respectively. To validate gene changes, expressions of twelve candidate genes related to germ cell development were examined by real-time PCR and found to be consistent with the of GeneChip data. Moreover, the immunostaining results suggested that ovarian development during different stages was regulated by different genes. Furthermore, a Raldh2 knockdown chicken model was produced to investigate the fundamental role of Raldh2 in meiosis initiation. It was found that meiosis occurred abnormally in Raldh2 knockdown ovaries, but the inhibitory effect on meiosis was reversed by the addition of exogenous RA. This study offers insights into the profile of gene expression and mechanisms regulating ovarian development, especially the notable role of Raldh2 in meiosis initiation in the chicken. © 2015 by the Society for Reproduction and Development. Source

Zhang Z.,Nanjing Agricultural UniversityJiangsu Province | Chen Z.,Nanjing Agricultural UniversityJiangsu Province | Hou Y.,Nanjing Agricultural UniversityJiangsu Province | Duan Y.,Nanjing Agricultural UniversityJiangsu Province | And 3 more authors.
Plant Disease | Year: 2015

Carbendazim, a methyl benzimidazole carbamate (MBC)-group fungicide, has been used to control rice bakanae disease, caused by Fusarium fujikuroi (teleomorph: Gibberella fujikuroi), for decades in China. Previous research revealed that point mutations (E198V, GAG to GTG at codon 198, and F200Y, TTC to TAC at codon 200) of the β2-tubulin gene conferred resistance of F. fujikuroi to MBC. In this study, primer-introduced restriction analysis polymerase chain reaction (PIRA-PCR) was developed to determine genotypes with resistance of F. fujikuroi to MBC. A PCR template of each strain was created by an outer primer pair. Fragments with 177 bp (for mutation at codon 235) and 146 bp (for E198V) were amplified by nested PCR, with two inner primer pairs designed and synthesized according to the nucleotide sequence of β2-tubulin for further enzyme digestion validation, respectively. AccII and PmaCI restriction enzyme recognition sites were introduced artificially by inner primers to differentiate MBC-sensitive and -resistant strains, respectively. The sensitivity of each strain to MBC was indirectly determined by analyzing electrophoresis patterns of the resulting amplified fragments after simultaneous digestion by both AccII and PmaCI. PIRA-PCR produced the same result as conventional methods in 6% of the time. PIRA-PCR is a sensitive and effective method for genotyping resistance alleles of F. fujikuroi strains to MBC. © 2015 The American Phytopathological Society. Source

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