Agri Gene最新文献

High-molecular-weight glutenin subunits (HMW-GS) are important in wheat end-use quality. We characterized a HMW-GS Dy10 mutant subunit with faster mobility than Dy10 in SDS-PAGE, and with equal mobility to Dy10 in urea-containing SDS-PAGE gel. A missense mutation was identified in the mutant subunit, which led to substitution of the 328th amino acid residue from ser (S) to phe (F) in the repetitive domain. The Dy10 subunit containing the missense mutation was designated as Dy10-m328SF (Genbank No. KU934284). Our results indicate that a single amino acid substitution in the repetitive domain can change the mobility of Dy10.

Complete mitochondrial genome sequences were assembled for the rice weevil, Sitophilus oryzae (17,602 bp) and the maize weevil, S. zeamais (18,105 bp; Coleoptera: Curculionidae: Dryophthorinae), which encode 13 protein coding genes (PCGs), 22 transfer RNAs, and 2 ribosomal RNAs that is typical among Animalia. The A + T-rich control regions of S. oryzae (2818 bp) and S. zeamais (2832 bp) are the longest described to date among curculionoidea (weevils). Additionally, non-coding intergenic regions have increased in size due to expansion of tandem repeat arrays, but is more pronounced in S. zeamais (606 bp) compared to S. oryzae (109 bp). A total of 69 and 22 substitution mutations were found among reads from S. oryzae and S. zeamais, out of which 68 and 2 were predicted in the PCGs respectively, with a majority in NADH Dehydrogenase subunit I. Phylogenetic analyses of coleopteran insects based upon full mitogenomes PCG sequence supported the existence of Curculionoidea and Tenebrionoidea as a monophyletic group, whereas the Cucujoidea and Elateroidea remain paraphyletic. The mitogenomes of these two Sitophilus species provide insight into short-term evolutionary relationships among curculionid beetles, and provide resources for the potential deciphering of more broad systematic questions in the Order Coleoptera.

The maternal recognition of pregnancy in pigs occurs around day 12–13 of gestation. During this period embryo-derived signals act on the endometrium and the myometrium to trigger tissue-specific responses that lead to proper implantation and maintenance of pregnancy. The present study utilized gene expression microarrays to examine embryo-induced alterations in porcine endometrial and myometrial transcriptome on days 12–13 of pregnancy. It has been confirmed that the endometrium and the myometrium, in response to embryonic signals, express unique set of genes. However, a key discovery of this study is the fact that genes that encode crucial factors involved in maternal recognition of pregnancy and preparation of uterus for implantation are regulated similarly in both tissues. Genes responsible for PGs synthesis (PTGES2, PTGR2), uterine development and remodeling (HOXA10, GRB10, MTOR) and innate immune response and immunomodulation (NFKBIA, TRAFD1) were regulated similarly in the endometrium and the myometrium. This suggests that in pigs these tissues act in sync to support embryos during peri-implantation.

The R2R3MYB proteins comprise one of the largest families of transcription factors and play regulatory roles in developmental processes and defense responses in plants. However, limited comprehensive genomic and functional analyses of these genes in eggplant (Solanum melongena L.) have been performed. In this study, 73 R2R3MYB genes were identified in the recently released eggplant genome and further classified into 20 subgroups based on the phylogenetic analysis of their complete protein sequences. Phylogenetic comparisons of this superfamily’s members among eggplant, tomato, Arabidopsis, grape, rice, poplar, soybean, cucumber and apple revealed that the putative functions of some of the eggplant R2R3MYB proteins were clustered into the Arabidopsis functional clades. The tissue specificity or differential expression of SmR2R3MYBs in different tissues suggested the differential regulation of tissue development as well as metabolism. The transcript abundance analysis under abiotic stress identified a group of R2R3MYB genes that responded to one or more treatments, suggesting that SmR2R3MYBs played major roles in the plants response to abiotic stress and were involved in signal transduction pathways. Genes related to anthocyanin synthesis were also identified. This study not only provides a solid foundation for the functional dissection of the eggplant R2R3MYB gene family, but may also be useful for the future synthesis of anthocyanins in eggplant.

Stilbene synthase plays important roles in the secondary metabolic pathways of many plants as polyketide synthase. Increased consumption of its metabolite, resveratrol, is highly beneficial to our health. The stilbene synthase 6 gene (VqSTS6) has been obtained from the Chinese wild Vitis quinquangularis accession ‘Danfeng-2’. Our previous studies have shown that this stilbene synthase gene exhibits higher levels of expression than other VqSTSs in the all six grape organs. To characterize gene expression, we cloned the promoter of VqSTS6 by homologous cloning. Alignment between the VqSTS6 gene sequence and Vitis vinifera VvSTS48 showed 99.24% identity. The promoter of VqSTS6 fused with the β-glucuronidase (GUS) gene was transformed into tomato (Solanum lycopersicum L. ‘Micro-Tom’) and the activity expression in four organs (roots, stems, leaves and fruits) was analyzed. This revealed that expression was much higher in the fruit than in the other three organs. We also used promoter deletion to show that the –518 bp to –411 bp region contained skn-1_motifs are the core functional promoter region in fruit development. These results evidence the special role of VqSTS6 promoter in regulating its expression in a high-resveratrol grapevine.

The aim of this study was to document the expression and localization of fibroblast growth factor (FGF) family members comprising of fibroblast growth factor (FGF1, FGF2, FGF7, FGF10), and their receptors (FGFR1, FGFR2, FGFR3, FGFR4, FGFR2IIIB, FGFR2IIIC) in buffalo corpus luteum (CL) obtained at different stages of the estrous cycle. In addition, the synergistic role of FGF2 and/or vascular endothelial growth factor (VEGF) on P₄ secretion and mRNA expression of steroidogenic acute regulatory protein (StAR), cytochrome P450 (CYP11A1), 3-beta-hydroxysteroid dehydrogenase (3βHSD), proliferating cell nuclear antigen (PCNA), BCL-2 associated X protein (BAX) and von willebrand factor (vWF) were studied in luteal cell culture obtained from mid-luteal phase (MLP) of estrous cycle in buffalo. Real-time PCR (qPCR), western blot, and immunohistochemistry were used to investigate mRNA and protein expressions, and the localization of examined factors whereas P₄ secretion was assessed by RIA. The mRNA and protein expression of FGF1 and FGFR1 were maximum (P < 0.05) during MLP whereas FGF2 was maximum (P < 0.05) during early luteal phase (ELP). FGF7, FGF10, FGFR2, FGFR3, FGFR4, FGFR2IIIb, and FGFR2IIIc mRNA and protein expression did not change among luteal phases. FGF family members were localized in cytoplasm of luteal cells as well as in endothelial cells. P₄ secretion in luteal cells treated with FGF2 or VEGF alone showed the maximum values (P < 0.05) with the highest dose at 72 h. P₄ secretion was found to be greater (P < 0.05) in luteal cells treated with FGF2 + VEGF compared to FGF2 or at 72 h of incubation. The mRNA expression of all factors were maximum (P < 0.05) whereas BAX was minimum (P < 0.05) at highest dose cultured for 72 h of luteal cells subjected with either protein alone or in combination. Summarizing, the present findings explore the synergistic role of FGF2 and VEGF on steroidogenesis, angiogenesis, cell viability through an autocrine and paracrine actions in buffalo CL.

Muscle growth rate is a critical issue for abalone aquaculture due to impacts on production costs. However, knowledge of the molecular mechanisms involved in molluscan myogenesis is limited. Therefore, the myostatin gene in the red abalone Haliotis rufescens (Hr-MSTN) was characterized and evaluated at the transcriptional level using RNA interference and gene silencing correlated with the insulin pathway as a proxy for somatic growth. Hr-MSTN cDNA was comprised of 2346 base pairs that encoded 489 amino acids and that contained structural characteristics typical of the transforming growth factor-β superfamily, including C-terminal signal peptide, propeptide domain, and transforming growth factor-β regions. Gene expression analysis revealed ubiquitous transcript expression in all tested red abalone tissues, but the muscle and mantle evidenced the highest transcriptional activity. RNA interference against MSTN mRNA significantly downregulated MSTN at 14 days post-injection, correlating with an upregulation of the insulin-related genes Insulin receptor, Growth factor receptor bound 2, and Proto-oncogene serine/threonine-protein kinase. These results suggest that MSTN silencing can promote activation of the insulin transcription pathway and consequently trigger somatic growth in the red abalone. This study is the first to evaluate the role of MSTN in gastropods using RNA interference, thus providing new perspectives for genetic improvement programs in abalone aquaculture.

Even though cultivated rice is highly sensitive to salinity, significant variability exists in the primary and secondary gene-pool of rice with respect to traits of salinity tolerance. Breeding salinity tolerance rice varieties is imperative due to climate change and increasing rice demand for global population. A meta-analysis of plethora of genomic data and published literature available on various genes/factors associated with response to rice salinity and tolerance can be used to enlist selected candidates genes affecting salinity. Such genes can be utilized to identify potential candidate salinity resistance genes from donor rice genotypes and facilitate their transfer to high yielding varieties of rice through marker-assisted breeding. This approach has tremendous advantage over transgenic approach as no bio-safety or regulatory issues are involved in exploiting the variability.

Meta-analyses were performed on three datasets viz., rice microarray data of 166 series comprising of 2586 samples, 1228 published research literature in the last one and half decades and RNA-Seq data of 454 and Illumina from Sequence Retrieval Archive (SRA) at NCBI. Among microarray dataset, six salinity related series were finally selected and multi experiment analysis revealed 2289 differentially expressed genes belonging to 44 gene families. Out of these, 13 families viz., AP2-EREBP, AUX/IAA, bZIP, C2H2, bHLH, C3H, HB, HSF, MYB, MYB-related, NAC, Tify and WRKY were selected. Applying various parameters on the published literature data, 13 genes were selected, of which five were common to the different microarray datasets. From RNA-Seq data, total of 751 differentially expressed genes were obtained from 21 gene families, out of which 11 genes were common with those obtained from microarray data and five genes, viz., AP2-EREBP/DREB, MYB, HSF, bZIP and NAC were common to all the three data sets. Based on the results obtained, a total of 31 meta-analyzed genes have been selected and recommended for use in genetic improvement programs aimed at salinity resistance in rice.

The meta-analysis of microarray, RNA-Seq and published literature has been successfully used to select 31 best salinity tolerance associated genes which can be exploited by candidate gene approach for targeted introgression through marker assisted breeding. This approach has multi-fold advantages, as it obviates statutory and ecological issues. Such endeavors are more warranted for combating the key abiotic stresses like salinity, whose effects are increasing due to a changing climate.

The spotted scat (Scatophagus argus) is a euryhaline marine fish with the capability to withstand different salinity levels and can tolerate abrupt transfer from salt water (SW) to fresh water (FW) or vice versa. Differentially expressed genes were identified by constructing suppression subtractive hybridization (SSH) cDNA libraries to explore branchial osmoregulatory mechanisms affected by salinity challenge. After trimming and blasting, 105 uniquely expressed sequence tags were identified. Among them, 24 candidate genes involved in the stress response, metabolism, and the respiratory chain were chosen for further validation by real-time polymerase chain reaction. More than two-fold expression differences were observed in ADP-ribosylation factor 1 (arf1), cyclic AMP-responsive element-binding protein 3-like 4 (creb3l4), EVI5-like protein (evi5l), protocadherin fat 2 (fat2), transferrin receptor protein 2 (tfr2), C-X-C chemokine receptor type 4 (cxcr4), aquaporin-3 (aqp3), sodium/potassium-transporting ATPase subunit beta-233 (nka β₂₃₃), serum and glucocorticoid-regulated kinase 1 (sgk1), and ras GTPase-activating-like protein IQGAP1 (iqgap1). Among these genes, aqp and nka are important osmoregulatory factors. AQP3 protein was observed being localized to the membranes of mitochondrion-rich cells (MRCs) and pillar cells of the gill of S. argus, and NKA β₂₃₃ subunit isoform was only found in MRC membranes by immunostaining. Significant differences in aqp3 and nka β₂₃₃ expression occurred within 24 h after being transferring into the freshwater, and nka β₂₃₃ expression level continued to highly upregulated 2 and 7 days post-transfer (dpt). These results suggest that branchial aqp3 and nka β₂₃₃ play important roles in response to hypo-osmotic stress in S. argus.

Transferrin (Tfa) plays an important role in iron regulation and is also known to be involved in response to infections. In this study, the full-length cDNA of the transferrin gene from Lutjanus argentiventris was cloned and characterized. The full-length cDNA of the tfa was 2482 bp encoding 690 amino acids containing an N-terminal signal peptide and the two conserved lobes. Tfa protein was highly similar to fish transferrins such as those from Larimichthys crocea (80%), Pagrus major (80%) and Dicentrarchus labrax (80%). Transferrin constitutive expression was found to be the highest in spleen and head-kidney. Moreover, the mRNA expression levels of tfa were measured by real-time PCR at 24 and 96 h in juveniles exposed to iron (Fe2x) during Aeromonas hydrophila infection. In general, the expression of tfa decreased in liver and intestine and increased in gill and skin in the iron group compared to control. The iron followed by Aeromonas hydrophila infection group caused a down-regulation in transferrin gene expression in all analyzed tissues at any point during the experiment. These findings demonstrate the evolutionary conservation of transferrin functions in vertebrates, involved in both the immune response and iron metabolism.