ZmC2H2-149 negatively regulates drought tolerance by repressing ZmHSD1 in maize.

Drought is a major abiotic stress that limits maize production worldwide. Therefore, it is of great importance to improve drought tolerance in crop plants for sustainable agriculture. In this study, we examined the roles of Cys2 /His2 zinc-finger-proteins (C2H2-ZFPs) in maize's drought tolerance as C2H2-ZFPs have been implicated for plant stress tolerance. By subjecting 150 Ac/Ds mutant lines to drought stress, we successfully identified a Ds-insertion mutant, zmc2h2-149, which shows increased tolerance to drought stress. Overexpression of ZmC2H2-149 in maize led to a decrease in both drought tolerance and crop yield. DAP-Seq, RNA-Seq, Y1H and LUC assays additionally showed that ZmC2H2-149 directly suppresses the expression of a positive drought tolerance regulator, ZmHSD1 (hydroxysteroid dehydrogenase 1). Consistently, the zmhsd1 mutants exhibited decreased drought tolerance and grain yield under water deficit conditions compared to their respective wild-type plants. Our findings thus demonstrated that ZmC2H2-149 can regulate ZmHSD1 for drought stress tolerance in maize, offering valuable theoretical and genetic resources for maize breeding programmes that aim for improving drought tolerance.

Structure and in vitro digestion characteristics of skim goat milk protein during processing: effects of fat separation.

BACKGROUND: Although nonfat milk has been used worldwide in the industrial dairy process, little is known about the effects of fat separation during the manufacturing process on skim milk's structural and digestive properties. This study investigated the effects of the manufacturing process on the structure and in vitro digestion properties of skim goat milk, particularly emphasizing fat separation. RESULTS: Changes in the surface charge and hydrophobicity of milk proteins caused by fat separation resulted in oxidation and aggregation in the subsequent homogenization, heat and spray-drying processing, which affected its digestibility. Compared with separation by dish separator (DS), skim milk after tubular centrifugal separation (CS) showed a higher initial and final digestibility. The CS samples also had a lower surface hydrophobicity level and higher free sulfhydryl content, ζ-potential, and average particle size (P < 0.05). Goat milk protein after CS was more readily oxidized and aggregated during the subsequent homogenization and heat treatment, as evidenced by the higher carbonyl content and particle size. Centrifugal separation also converted more ß-sheets to α-helices, thus promoting the aggregation of oxidized skim milk protein. CONCLUSION: The skim milk after CS and DS demonstrated different structural and digestive properties. Skim goat milk products after CS were more susceptible to oxidant-induced protein structural changes, resulting in higher protein digestibility. These findings provide insights into the mechanism involved in the control of gastric digestion of skim milk during manufacturing process. © 2023 Society of Chemical Industry.

Identification, functional characterization and expression pattern of myeloid differentiation factor 88 (MyD88) in Sepiella japonica.

Myeloid differentiation factor 88 (MyD88) is an adaptor protein involved in the interleukin-1 receptor and Toll-like receptor-induced activation of nuclear factor-κB (NF-κB). In this study a novel isoform of MyD88 in Sepiella japonica (SjMyD88) was cloned and functionally characterized (GenBank accession no. AQY56781.1). The complete cDNA sequence of SjMyD88 was 1912 bp and contained a 1017 bp open reading frame encoding 338 amino acid residues, which was similar to its mollusk orthologues in the length. BLASTp analysis suggested the deduced amino acids sequence of SjMyD88 shared high identity to the known MyD88, for instance, 64% identity with Octopus bimaculoides. Sequence analysis revealed two conserved domains, the N-terminal DD and the C-terminal TIR domain appeared in SjMyD88, which was consistent with MyD88 proteins from other species. The fusion expression of SjMyD88 and green fluorescent protein (EGFP) in HEK293 cells was conducted and cytoplasm localization was detected. Meanwhile, the TIR-pmCherry fusion protein showed red fluorescence and mainly distributed in the cytoplasm. After cotransfection MyD88-EGFP and TIR-pmCherry red obviously overlapped and changed to yellowish green. The results suggested that there was the interaction between homologous TIR-pmcherry and MyD88-EGFP. Tissues expression profiles analysis showed that SjMyD88 ubiquitously expressed in all tested tissues with the highest expression in the gills and livers except reproductive related tissue, and it was significantly induced in livers under LPS stress. These data provide insight into the roles of SjMyD88 in the TLR signaling pathway of S. japonica in response to pathogenic bacteria.

The expression of superoxide dismutase in Mytilus coruscus under various stressors.

Superoxide dismutases (SODs), a by-product of antioxidative defence system, protects organisms for eliminating excess reactive oxygen species (ROS) and maintaining the redox balance of immune system. The complete open reading frames (ORFs) of Cu/Zn-SOD and Mn-SOD were identified from Mytilus coruscus (designated as McSOD and MnSOD) by homologous cloning. The sequence lengths were 474bp and 687bp, encoding 157 and 228 amino acids respectively. The deduced amino acid sequences of McSOD and MnSOD shared high identities with Cu/Zn-SOD and Mn-SOD from other mollusca. The distributions of McSOD and MnSOD were detected in six tissues including adductor, hemocyte, gill, gonad, mantle and hepatopancreas, and the highest expressions were both in gills. The temporal expression of McSOD and MnSOD were up-regulated in gills under a variety of stress factors, including Vibrio parahemolyticus, Aeromonas hydrophila, Cu2+ and Pb2+. After being challenged with V. Parahemolyticus, the expressions of McSOD and MnSOD were increased rapidly at the initial hours, reaching the peaks of 4.9-fold and 15.3-fold respectively, and got to the highest levels of 43.5-fold and 7.1-fold after being challenged with A. hydrophila. The highest point of McSOD mRNA appeared at 15 d after being exposed to copper (7-fold at 0.5 mg/L and 13.2-fold at 1.5 mg/L), except for 0.1 mg/L group of Cu2+ maintaining to the normal level, but plumbum at 1 d (2.4-fold at 1.0 mg/L and 4.4-fold at 3.0 mg/L) and at 15 d (2.1-fold at 0.2 mg/L). The temporal expression peaks of MnSOD appeared differently after exposing to copper of various concentrations (0.1 mg/L at 10 d with 4.7-fold, 0.5 mg/L at 1 d with 17.9-fold and 1.5 mg/L at 3 d with 13.2-fold). Whereas in plumbum exposing treatments, the 3.0 mg/L group jumped to the peak at 1 d (18.2-fold), the 0.2 mg/L and 1.0 mg/L groups had little change and maintained at the normal level throughout the experiment. The results provided several new evidences for further understanding of the regulatory mechanism of SOD on the innate immune system in bivalve.

Ultrasound improves the thermal stability and binding capacity of ovomucin by promoting the dissociation of insoluble ovomucin aggregates.

Ovomucin (OVM) is a natural glycoprotein with various biological activities but poor solubility. This study aimed to enhance the solubility of OVM by using an ultrasonic-assisted method. The effect of ultrasound (US) on the structure, thermal stability and biological functions of OVM aggregates was evaluated. It was found that insoluble OVM aggregates were dissociated and the solubility increased significantly to 90.0 % after US under 400 W for 45 min. US also improved the onset temperature (To) and denaturation temperature (Td) of OVM. More importantly, the cholesterol binding capacity of both OVM and its digestion products were significantly improved after US (p < 0.05). The gastrointestinal digestion products of US-OVM also showed higher α-amylase and α-glucosidase inhibition than native OVM aggregates. US-induced dissociation of OVM aggregates and the conversion of ß-sheet and ß-turn to random coil, resulting in the exposure of hydrophobic binding sites may be an important reason for the enhanced stability and adsorption capacity. These findings suggested that US was an effective method for preparing soluble OVM and improved its adsorption capacity, which can further facilitate the application of OVM in the food industry.

ZmDWF1 regulates leaf angle in maize.

Leaf angle (LA) is a critical agronomic trait enhancing grain yield under high-density planting in maize. A number of researches have been conducted in recent years to investigate the quantitative trait loci/genes responsible for LA variation, while only a few genes were identified through map-based cloning. Here we cloned the ZmDWF1 gene, which was previously reported to encode Δ24-sterol reductase in the brassinosteroids (BRs) biosynthesis pathway. Overexpression of ZmDWF1 resulted in enlarged LA, indicating that ZmDWF1 is a positive regulator of LA in maize. To reveal the regulatory framework of ZmDWF1, we conducted RNA-Sequencing and yeast-two hybrid (Y2H) screening analysis. RNA-Sequencing analyzing results indicate ZmDWF1 mainly affected expression level of genes involved in cell wall associated metabolism and hormone metabolism including BR, gibberellin, and auxin. Y2H screening with Bi-FC assay confirmed three proteins (ZmPP2C-1, ZmROF1, and ZmTWD1) interacting with ZmDWF1. We revealed a new regulatory network of ZmDWF1 gene in controlling plant architecture in maize.

A Nucleoporin NUP58 modulates responses to drought and salt stress in maize (Zea mays L.).

Nuclear pore complex (NUP) is the main transport channel between cytoplasm and nucleoplasm, which plays an important role in stress response. The function of NUPs was widely reported in yeast and vertebrate but rarely in plants. Here, we identified a nuclear pore complex (ZmNUP58), that is tightly associated with drought and salt tolerance phenotype accompanied with phenotypic and physiological changes under drought and salt stress. The overexpression of ZmNUP58 in maize (Zea mays L.) significantly promotes both chlorophyll content and activities of antioxidant enzymes under drought- and salt-stressed conditions. RNA-Seq analysis showed that ZmNUP58 could regulate the expression of genes related to phytohormone synthesis and signaling, osmotic adjustment substances, antioxidant enzyme system, cell wall biosynthesis, glucose metabolism and aquaporin. The results provide novel insights into the regulatory role of ZmNUP58 in improving drought and salt tolerance through regulating phytohormone and other stress response genes in maize.

A novel biomarker for marine environmental pollution of CAT from Mytilus coruscus.

Bivalves use anti-oxidative enzyme systems to defend themselves against excessive reactive oxygen species, which are often catalyzed by environmental pollution. As a key member of anti-oxidative enzyme family, catalase plays a crucial role in scavenging the high level of reactive oxygen species to protect organisms against various oxidative stresses. In this study, a catalase homologue was identified from Mytilus coruscus (named McCAT, KX957929). The open reading frame of McCAT was 1844bp with a 5' untranslated region of 341bp and a 3' untranslated region of 927bp. The deduced amino acid sequence was 512 residues in length with theoretical pI/MW 8.02/57.91kDa. BLASTn and phylogenetic analyses strongly suggested that it was a member of catalase, also known as CAT family for its conserved catalytic site motif and proximal heme-ligand signature motif. Real-time fluorescence quantitative PCR showed that constitutive expression of McCAT was occurred, with increasing order in mantle, adductor, gill, hemocyte, gonad and hepatopancreas. It was observed that bacterial infection and heavy metals stimulation up-regulated McCAT mRNA expression in hepatopancreas with time-dependent manners. The maximum expression appeared at 8h after pathogenic bacteria injecting, with 15-fold in Vibrio parahemolyticus and 60-fold in Aeromonas hydrophila than that of 0h. The highest point of McCAT mRNA appeared at different times for exposure to heavy metals with copper at day 5 (0.1mg/L 30-fold, 0.5mg/L 15-fold, 1.5mg/L 6-fold) and plumbum at day 3 (3.0mg/L 20-fold). The enzymatic activity analysis found that McCAT activity in the gill of M. coruscus was affected by heavy metals concentration. The results suggested that McCAT plays a significant role in antioxidation and the expression of McCAT can be used as a biomarker for detection of marine environmental pollution.