High-fibre diets regulate antioxidative capacity and promote intestinal health by regulating bacterial microbiota in growing pigs.

This experiment was conducted to investigate the effects of a high-fibre diet on growth performance, nutrients digestibility, intestinal health, and intestinal microbiota composition of growing pigs. Twelve healthy "Duroc × Landrace × Yorkshire" castrates (49 ± 1.35 kg) were randomly divided into two groups with six replicates and one pig per replicate. The two diet treatments were fed the basal diet (CON) based on corn and soybean meal and high fibre diet (HF) respectively. The nutritional levels of the two treatments were the same. The experiment lasted 28 days. The results showed that the addition of 16% wheat bran fibre to the diet of growing pigs did not affect growth performance (p > 0.05). Compared with the CON, contents of isobutyric and butyric acid, GSH-PX and T-AOC in serum were increased in the HF. It decreased the gross energy digestibility and acetic acid content in feces of growing pigs (p < 0.05), the contents of GSH-PX and T-AOC in serum. It decreased the gross energy digestibility and acetic acid content in feces of growing pigs (p < 0.05). Compared with the CON, the Shannon, and Chao1 indexes of the HF were increased (p < 0.05). At the phylum level, the abundance of g_Lactobacillus increased in the HF (p < 0.05). Correlation analysis showed that a total of 18 microbial genera were correlated with antioxidant capacity and volatile fatty acid levels (p < 0.05). In summary, this study showed that adding 16% wheat bran to the diet of growing pigs had no effect on growth performance but helped to improve the richness and stability of intestinal microbiota, promote posterior intestinal fermentation and increase serum antioxidant capacity.

Effects of dietary supplementation of different levels of gamma-aminobutyric acid on reproductive performance, glucose intolerance, and placental development of gilts.

This study aimed to investigate the effects of dietary gamma-aminobutyric acid (GABA) supplementation on reproductive performance, glucose intolerance, and placental development of gilts during mid-late gestation. Based on the principle of backfat thickness consistency, 124 gilts at 65 d of gestation were assigned to three dietary groups: CON (basic diet, n = 41), LGABA (basic diet supplemented with 0.03% GABA, n = 42), and HGABA (basic diet supplemented with 0.06% GABA, n = 41). The litter performance, glucose tolerance, placental angiogenesis, and nutrients transporters were assessed. The LGABA group improved piglet vitality and placental efficiency and decreased area under the curve of glucose tolerance test compared to the CON group (P < 0.05). Meanwhile, the LGABA group enhanced placental vessel density, platelet endothelial cell adhesion molecule-1 levels and gene expression of fibroblast growth factor 18 (P < 0.05). Furthermore, LGABA showed an uptrend in glucose transporter type 1 mRNA level (P = 0.09). Taken together, this study revealed that the dietary supplementation of 0.03% GABA can improve piglet vitality, glucose intolerance, and placental development of gilts.

Glucose homeostasis and placental development are two key factors influencing reproductive performance of sows. Some studies have reported that gamma-aminobutyric acid (GABA) can improve glucose intolerance and cerebral angiogenesis in mice. Therefore, we hypothesized that GABA can improve reproductive performance, glucose intolerance, and placental development of gilts during mid-late gestation. In this study, gilts were randomly assigned into three groups: CON (basal diet), LGABA (basal diet supplemented with 0.03% GABA), and HGABA (basal diet supplemented with 0.06% GABA). Results showed that the LGABA group significantly improved the piglet viability, glucose intolerance, and placental development compared with the CON group. Therefore, GABA has a good prospect as a feed additive for gilts.

Chronic jet lag alters gut microbiome and mycobiome and promotes the progression of MAFLD in HFHFD-fed mice.

Metabolic dysfunction-associated fatty liver disease (MAFLD) is the most common chronic liver disease worldwide. Circadian disruptors, such as chronic jet lag (CJ), may be new risk factors for MAFLD development. However, the roles of CJ on MAFLD are insufficiently understood, with mechanisms remaining elusive. Studies suggest a link between gut microbiome dysbiosis and MAFLD, but most of the studies are mainly focused on gut bacteria, ignoring other components of gut microbes, such as gut fungi (mycobiome), and few studies have addressed the rhythm of the gut fungi. This study explored the effects of CJ on MAFLD and its related microbiotic and mycobiotic mechanisms in mice fed a high fat and high fructose diet (HFHFD). Forty-eight C57BL6J male mice were divided into four groups: mice on a normal diet exposed to a normal circadian cycle (ND-NC), mice on a normal diet subjected to CJ (ND-CJ), mice on a HFHFD exposed to a normal circadian cycle (HFHFD-NC), and mice on a HFHFD subjected to CJ (HFHFD-CJ). After 16 weeks, the composition and rhythm of microbiota and mycobiome in colon contents were compared among groups. The results showed that CJ exacerbated hepatic steatohepatitis in the HFHFD-fed mice. Compared with HFHFD-NC mice, HFHFD-CJ mice had increases in Aspergillus, Blumeria and lower abundances of Akkermansia, Lactococcus, Prevotella, Clostridium, Bifidobacterium, Wickerhamomyces, and Saccharomycopsis genera. The fungi-bacterial interaction network became more complex after HFHFD and/or CJ interventions. The study revealed that CJ altered the composition and structure of the gut bacteria and fungi, disrupted the rhythmic oscillation of the gut microbiota and mycobiome, affected interactions among the gut microbiome, and promoted the progression of MAFLD in HFHFD mice.

S-alkyl Dithiocarbamates Synthesis through Electrochemical Multicomponent Reaction of Thiols, Hydrogen Sulfide, and Isocyanides.

Dithiocarbamates synthesis is extremely important in plenty of biomedical and agrochemical applications, especially fungicide development, but remains a great challenge. In this work, we have successfully developed a multicomponent reaction protocol to convert H2S into S-alkyl dithiocarbamates under constant current conditions. No additional oxidants nor additional catalysts are required, and due to mild conditions, the reactions display a broad substrate scope, including varieties of thiols or disulfides.

Effects of ß-alanine on intestinal development and immune performance of weaned piglets.

Beta-alanine is an important amino acid involved in several metabolic reactions in the body. The study aimed to investigate the effect of ß-alanine supplementation on intestinal development and the immune performance of weaned piglets. Thirty-two 21-day-old healthy weaned piglets (half female and half male; Duroc × Landrace × Yorkshire) with an initial body weight of 8.11 ± 0.21 kg were randomly divided into 4 groups with 8 replicates of 1 pig each. The control group was fed a basal diet and the three experimental treatment groups were fed diets supplemented with 300, 600 and 1,200 mg/kg ß-alanine, respectively. The trial lasted 28 days and the diets fed were divided into 2 phases: the late lactation period (day 1 to 14) and the nursery period (day 15 to 28), during which the weaned piglets had free access to food and water. The regulatory effects of ß-alanine were further investigated in vitro using organoids obtained from the jejunum of piglets. In vivo, the addition of ß-alanine to the diet had no significant effect on the growth performance of weaned piglets (P > 0.05), but significantly reduced serum levels of immunoglobulin G (IgG) (P < 0.01), immunoglobulin M (IgM) (P = 0.005), and complement 3 (C3) (P = 0.017). The serum interleukin- 6 (IL-6) levels (P < 0.01) were significantly reduced in the 1,200 mg/kg treatment group. The addition of ß-alanine increased ileal villus height, with the most significant effect at a concentration of 300 mg/kg (P = 0.041). The addition of 600 mg/kg ß-alanine significantly up-regulated the expression of superoxide dismutase (SOD) activity (P = 0.020) and the zonula occludens-1 (ZO-1) gene (P = 0.049) in the jejunum. Diets supplemented with 300 mg/kg ß-alanine significantly increased the number of Ki67 positive cells in the jejunal crypts (P < 0.01). In vitro, ß-alanine increased the organoid budding rates (P = 0.001) and the budding height of the crypt significantly (P = 0.004). In conclusion, ß-alanine can improve intestinal morphology and barrier function, reduce inflammatory responses and alleviate the adverse effects of weaning stress on piglet intestinal health.

Effects of different starch diets on growth performance, intestinal health and faecal microbiota of growing pigs.

This experiment was conducted to investigate the effects of different starch source diets on growth performance, intestinal health, and, microbiota of growing pigs. Eighteen healthy "Duroc × Landrace × Yorkshire" pigs (50 ± 0.61 kg, Castrated boar) were randomly divided into three groups with six replicates and one pig per replicate. The pigs in the three treatments were fed diets prepared with cassava flour (CF), rice bran (RB) and sorghum flour (SF), respectively, and the nutritional levels of the three treatments were the same. The experiment lasted for 28 days. The results showed that pigs in the RB group had significantly increased average daily gain (ADG, p < 0.05) compared with pigs in CF and SF groups. Compared with pigs in the CF group, the final body weight (FBW) of growing pigs in the RB group was increased and the ratio of feed to gain (F: G) was decreased (p < 0.05). There was no significant difference between FBW and F: G between the SF group and the other two groups. Compared with the CF group, the RB group significantly increased the jejunum amylase activity (p < 0.05), and there was no significant difference between the SF group and the other two groups. Compared with growing pigs in the CF group and SF group, the duodenal villus height and villus height/crypt depth ratio of growing pigs in the RB group were significantly increased (p < 0.05). The concentrations of acetic acid, propionic acid, and total VFA in the colon and caecum of piglets in the SF group were significantly increased (p < 0.05) compared to piglets in CF and RB groups, and there was no significant difference between the CF group and RB group. Compared with the RB group, caecal butyric acid concentration was significantly increased in SF and CF groups (p < 0.05). Seven dominant phyla were identified at the phylum level, among which Firmicutes, Bacteroidota and Spirochaetota were dominant phyla, accounting for 74.18%, 14.87% and 6.56% of the RB group respectively. Cassava flour group accounted for 80.22%, 9.64% and 3.71%; Accounting for 65.33%, 17.34% and 13.07% of the SF group. Through the comparative analysis of microbial differences among the treatment groups, it was found that at the phylum level, compared with the SF group, the abundance of Synergistota in the diet of the CF group and the diet of the RB group was significantly increased (p < 0.05). The abundance decreased significantly (p < 0.05). The quantity of Desulfobacterota in the RB group was significantly higher than that in the CF group (p < 0.05). In conclusion, compared with sorghum starch and cassava starch, RB starch can improve the activity of digestive enzymes and villus height in the small intestine of growing pigs and promote the growth of pigs by protecting the intestinal health of growing pigs.

Effects of dietary ß-alanine supplementation on growth performance, meat quality, carnosine content, amino acid composition and muscular antioxidant capacity in Chinese indigenous Ningxiang pig.

ß-alanine has been demonstrated to improve carcass traits and meat quality of animals. However, no research has been found on the effects of dietary ß-alanine in the meat quality control of finishing pigs, which are among the research focus. Therefore, this study aimed to evaluate the effects of dietary ß-alanine supplementation on growth performance, meat quality, carnosine content, amino acid composition and muscular antioxidant capacity of Chinese indigenous Ningxiang pigs. The treatments contained a basal diet (control, CON) and a basal diet supplemented with 600 mg/kg ß-alanine. Each treatment group consisted of five pens, with five pigs per pen. Results showed that compared with CON, supplemental ß-alanine did not affect the final body weight, average daily gain, average daily feed intake and the feed-to-gain ratio of pigs. Dietary ß-alanine supplementation tended to increase the pH45 min (p = 0.071) while decreasing the shear force (p = 0.085) and the drip loss (p = 0.091). Moreover, it improved (p < 0.05) the activities of glutathione peroxidase and catalase and lessened (p < 0.05) malondialdehyde concentration. Added ß-alanine in diets of finishing pigs could enhance the concentrations of arginine, alanine, and glutamate (p < 0.05) in the longissimus dorsi muscle and tended to raise the levels of cysteine, glycine and anserine (p = 0.060, p = 0.098 and p = 0.091 respectively). Taken together, our results showed that dietary ß-alanine supplementation contributed to the improvement of the carcass traits, meat quality and anserine content, the amelioration of muscle antioxidant capacity and the regulation of amino acid composition in Chinese indigenous Ningxiang pigs.

Effects of low protein diet with a balanced amino acid pattern on growth performance, meat quality and cecal microflora of finishing pigs.

BACKGROUND: The present study aimed to investigate the effects of low protein diets balanced with four amino acids on growth performance, meat quality and cecal microflora of finishing pigs. Fifty-four healthy hybrid barrows (Duroc × Landrace × Yorkshire) with an average body weight of 70.12 ± 4.03 kg were randomly assigned to one of the three dietary treatments with six replicate pens per treatment (three barrows per pen). The three dietary treatments included a normal protein diet (NP), a low protein diet (LP) and a very low protein diet (VLP). RESULTS: The average daily gain, average daily feed intake and feed conversion ratio of pigs were not significantly changed with the LP and VLP diets compared to the NP diet (P > 0.05). The water holding capacity and shear force of longissimus dorsi muscle were decreased, whereas the intramuscular fat content of the longissimus dorsi muscle was increased (P < 0.05) in pigs fed with the LP and VLP diets compared to the NP diet. The contents of saturated fatty acids in muscle were decreased (P < 0.05), whereas the content of polyunsaturated fatty acids in muscle was increased (P < 0.01) with the VLP diet compared to the NP diet. The contents of histamine, spermidine, spermine and tyramine of muscle were decreased with the VLP diet compared to the NP diet (P < 0.05). The relative abundance of Turicibacter, Terrisporobacter, Clostridium_sensu_stricto_1 and UCG-005 was higher (P < 0.05), whereas the relative abundance of Lactobacillus and Streptococcus was lower (P < 0.05) in pigs fed with the LP and VLP diets compared to the NP diet. Based on the correlation of cecal microbiota and cecal biogenic amine, the contents of tyramine, spermidine and histamine were negatively correlated with the abundance of Terrisporobacter (P < 0.01) and the content of histamine was positively correlated with the abundance of Lactobacillus (P < 0.01). CONCLUSION: Balanced with four essential amino acids, the VLP diet with crude protein levels decreased by > 4% increased the intramuscular fat content, changed the fatty acid and amino acid composition of longissimus dorsi muscle and the profile of cecum microbiota, and reduced the content of cecum bioamine, with no negative effect on the growth performance of pigs. © 2022 Society of Chemical Industry.

Effect of Different Levels of Niacin on Serum Biochemical Parameters, Antioxidant Status, Cytokine Levels, Inflammatory Gene Expression and Colonic Microbial Composition in Weaned Piglets.

Niacin plays an important role in regulating the gut health of weaned piglets. In this study, 48 25-day-old weaned piglets (7.9 ± 0.20 kg) produced by 14 sows (3 to 4 piglets per sow) were randomly divided into 4 groups with 6 replicates in each group and 2 piglets in each replicate. Each group was fed diets supplemented with 22.5 (N1), 30 (N2), 45 (N3), and 75 (N4) mg/kg of niacin, respectively. Samples were taken at 7 and 14 d, respectively. The study shows that changes in niacin levels significantly affected the content of IgG and IgM in the serum (p < 0.05). Niacin had a significant effect on antioxidant parameters such as MDA, T-SOD, and CuZn-SOD in the jejunal mucosa of weaned piglets (p < 0.05). Moreover, significant differences were observed in the expression of cytokines such as TGF-ß, TNF-α, and COX2 in the jejunal mucosa (p < 0.05). The 16S rRNA sequencing analysis showed that there were significant differences in the colonic species composition, which were also accompanied by changes in the isovaleric acid content (p < 0.05). In conclusion, an appropriate increase in niacin dose based on NRC (2012) has an important role in improving the antioxidant status of weaned piglets, alleviating intestinal inflammation in piglets, improving immunity, and regulating the structure of the microbiota.

Higher niacin intakes improve the lean meat rate of Ningxiang pigs by regulating lipid metabolism and gut microbiota.

As one of the local pig breeds in China with a high fat rate, improving the lean meat rate of Ningxiang pigs through nutritional intervention is an urgent issue to be solved. As an important feed additive, niacin plays an important role in lipid metabolism. The purpose of this study was to investigate the regulation and mechanism of niacin on fat deposition in Ningxiang pigs. Thirty-four Ningxiang pigs (53.34 ± 2.78 kg) were randomly divided into two groups with five replicates each, with three to four Ningxiang pigs per replicate. The control group was fed a basal diet (contained 22 mg/kg niacin), and the experimental group was fed the same diet supplemented with an additional 100 mg/kg of niacin. The experimental period lasted 60 days. One Ningxiang pig was selected for slaughter sampling for each replicate. This study found that lean meat percentage of Ningxiang pigs in the experimental group was significantly increased (P < 0.05), accompanied by a significant decrease in fat percentage (P < 0.05). 16S rRNA sequencing analysis found an abundance of Streptococcus in the experimental group (P < 0.05), along with significantly decreased levels of Lactobacillus (P < 0.05). The changes in some OTUs belonging to Firmicutes, Bacteroidota, and Actinobacteriota were closely related to the changes in the fat rate and lean meat rate of Ningxiang pigs (P < 0.05). LC-MS metabolomics analysis found that about 43.75% of the differential metabolites were related to lipids and lipid-like molecules in the liver (P < 0.05). Spearman's correlation analysis showed correlations between the carcass traits, microbiota, and liver metabolites. In conclusion, niacin improves lean meat percentage and reduces fat deposition by regulating lipid metabolism and gut microbiota composition in Ningxiang pigs.

BREVIPEDICELLUS and ERECTA control the expression of AtPRX17 to prevent Arabidopsis callus browning.

Efficient in vitro callus generation is required for tissue culture propagation, a process that allows for plant regeneration and transgenic breeding for desired phenotypes. Identifying genes and regulatory elements that prevent impaired callus growth and callus browning is essential for the development of in vitro callus systems. Here, we show that the BREVIPEDICELLUS and ERECTA pathways in Arabidopsis calli converge to prevent callus browning, and positively regulate the expression of the isoperoxidase gene AtPRX17 in rapidly growing calli. Loss-of-function mutations in both BREVIPEDICELLUS and ERECTA resulted in markedly increased callus browning. Transgenic lines expressing 35S pro::AtPRX17 in the bp-5 er105 double mutant background fully rescued this phenotypic abnormality. Using in vivo (chromatin immunoprecipitation-PCR and transient expression) and in vitro (electrophoretic mobility shift assays) assays, we observed that the BREVIPEDICELLUS protein binds directly to the upstream sequence of AtPRX17 to promote its transcription during callus growth. ERECTA is a ubiquitous factor required for cell proliferation and growth. We show that ERECTA positively regulates the expression of the transcription factor WRKY6, which directly binds to a separate site on the AtPRX17 promoter, further increasing its expression. Our data reveal an important molecular mechanism involved in the regulation of peroxidase isozyme expression to reduce Arabidopsis callus browning.

The mutated Bacillus amyloliquefaciens strain shows high resistance to Aeromonas hydrophila and Aeromonas veronii in grass carp.

Bacillus amyloliquefaciens X030 (BaX030) has broad-spectrum antibacterial activity against the fish pathogens Aeromonas hydrophila and Aeromonas veronii. To improve its antibacterial effect, BaX030 was subjected to compound mutagenesis of atmospheric and room temperature plasma (ARTP) and nitrosoguanidine (NTG). The results showed that, compared with the original strain, the production of macrolactin A and oxydifficidin in mutated strain N-11 increased to 39 % and 268 %, respectively. The re-sequencing analysis suggested that there were SNPs and InDels in the gene clusters focused on the sucrose utilization pathway, glycolysis pathway and fatty acid synthesis pathway. Scanning electron microscopy revealed that strain N-11 became thin and long. The qRT-PCR results indicated that the expression of immune factors in the liver or kidney tissue of grass carp increased after feeding with N-11. H&E staining and protection experiments also showed that the mortality and surface symptoms of grass carp infected by the two pathogens were significantly reduced. The study identified a probiotic strain with potential application value in aquaculture production and provided a new strategy for the discovery of new strains with higher antibacterial biological activity.

iTRAQ analysis reveals the effect of gabD and sucA gene knockouts on lysine metabolism and crystal protein formation in Bacillus thuringiensis.

Lysine metabolism plays an important role in the formation of the insecticidal crystal proteins of Bacillus thuringiensis (Bt). The genes lam, gabD and sucA encode three key enzymes of the lysine metabolic pathway in Bt4.0718. The lam gene mainly affects the cell growth at stable period, negligibly affected sporulation and insecticidal crystal protein (ICP) production. While, the deletion mutant strains of the gabD and sucA genes showed that the growth, sporulation and crystal protein formation were inhibited, cells became slender, and insecticidal activity was significantly reduced. iTRAQ proteomics and qRT-PCR used to analyse the differentially expressed protein (DEP) between the two mutant strains and the wild type strain. The functions of DEPs were visualized and statistically classified, which affect bacterial growth and metabolism by regulating biological metabolism pathways: the major carbon metabolism pathways, amino acid metabolism, oxidative phosphorylation pathways, nucleic acid metabolism, fatty acid synthesis and peptidoglycan synthesis. The gabD and sucA genes in lysine metabolic pathway are closely related to the sporulation and crystal proteins formation. The effects of DEPs and functional genes on basic cellular metabolic pathways were studied to provide new strategies for the construction of highly virulent insecticidal strains, the targeted transformation of functional genes.

Pediococcus pentosaceus: Screening and Application as Probiotics in Food Processing.

Lactic acid bacteria (LAB) are vital probiotics in the food processing industry, which are widely spread in food additives and products, such as meat, milk, and vegetables. Pediococcus pentosaceus (P. pentosaceus), as a kind of LAB, has numerous probiotic effects, mainly including antioxidant, cholesterol-lowering, and immune effects. Recently, the applications in the probiotic- fermentation products have attracted progressively more attentions. However, it is necessary to screen P. pentosaceus with abundant functions from diverse sources due to the limitation about the source and species of P. pentosaceus. This review summarized the screening methods of P. pentosaceus and the exploration methods of probiotic functions in combination with the case study. The screening methods included primary screening and rescreening including gastric acidity resistance, bile resistance, adhesion, antibacterial effects, etc. The application and development prospects of P. pentosaceus were described in detail, and the shortcomings in the practical application of P. pentosaceus were evaluated to make better application of P. pentosaceus in the future.

Transcriptomic Analysis of the Interaction Between FLOWERING LOCUS T Induction and Photoperiodic Signaling in Response to Spaceflight.

Spaceflight has an impact on the growth and development of higher plants at both the vegetative stage and reproductive stage. A great deal of information has been available on the vegetative stage in space, but relatively little is known about the influence of spaceflight on plants at the reproductive stage. In this study, we constructed transgenic Arabidopsis thaliana plants expressing the flowering control gene, FLOWERING LOCUS T (FT), together with the green fluorescent protein gene (GFP) under control of a heat shock-inducible promoter (HSP17.4), by which we induced FT expression inflight through remote controlling heat shock (HS) treatment. Inflight photography data showed that induction of FT expression in transgenic plants in space under non-inductive short-day conditions could promote flowering and reduce the length of the inflorescence stem in comparison with that of wild-type plants under the same conditions. Whole-genome microarray analysis of gene expression changes in leaves of wild-type and these transgenic plants grown under the long-day and short-day photoperiod conditions in space indicated that the function of the photoperiod-related spaceflight responsive genes is mainly involved in protein synthesis and post-translation protein modulation, notably protein phosphorylation. In addition, changes of the circadian component of gene expression in response to spaceflight under different photoperiods indicated that roles of the circadian oscillator could act as integrators of spaceflight response and photoperiodic signals in Arabidopsis plants grown in space.

Molecular Basis to Integrate Microgravity Signals into the Photoperiodic Flowering Pathway in Arabidopsis thaliana under Spaceflight Condition.

Understanding the effects of spaceflight on plant flowering regulation is important to setup a life support system for long-term human space exploration. However, the way in which plant flowering is affected by spaceflight remains unclear. Here, we present results from our latest space experiments on the Chinese spacelab Tiangong-2, in which Arabidopsis wild-type and transgenic plants pFT::GFP germinated and grew as normally as their controls on the ground, but the floral initiation under the long-day condition in space was about 20 days later than their controls on the ground. Time-course series of digital images of pFT::GFP plants showed that the expression rhythm of FT in space did not change, but the peak appeared later in comparison with those of their controls on the ground. Whole-genome microarray analysis revealed that approximately 16% of Arabidopsis genes at the flowering stage changed their transcript levels under spaceflight conditions in comparison with their controls on the ground. The GO terms were enriched in DEGs with up-regulation of the response to temperature, wounding, and protein stabilization and down-regulation of the function in circadian rhythm, gibberellins, and mRNA processes. FT and SOC1 could act as hubs to integrate spaceflight stress signals into the photoperiodic flowering pathway in Arabidopsis in space.

Enhancing the insecticidal activity of new Bacillus thuringiensis X023 by copper ions.

BACKGROUND: A new Bacillus thuringiensis X023 (BtX023) with high insecticidal activity was isolated in Hunan Province, China. The addition of metals (Cu, Fe, Mg and Mn) to the medium could influence the formation of spores and/or insecticidal crystal proteins (ICPs). In previous studies, Cu ions considerably increased the synthesis of ICPs by enhancing the synthesis of poly-ß-hydroxy butyrate. However, the present study could provide new insights into the function of Cu ions in ICPs. RESULTS: Bioassay results showed that wild strain BtX023 exhibited high insecticidal activity against Plutella xylostella. The addition of 1 × 10-5 M Cu2+ could considerably increase the expression of cry1Ac and vip3Aa, and the insecticidal activity was enhanced. Quantitative real-time polymerase chain reaction (qRT-PCR) and proteomic analyses revealed that the upregulated proteins included amino acid synthesis, the glyoxylate pathway, oxidative phosphorylation, and poly-ß-hydroxy butyrate synthesis. The Cu ions enhanced energy metabolism and primary amino acid synthesis, will providing abundant raw material accumulation for ICP synthesis. CONCLUSION: The new strain BtX023 exerted a strong insecticidal effect on P. xylostella by producing ICPs. The addition of 1 × 10-5 M Cu2+ in the medium could considerably enhance the expression of the cry1Ac and vip3Aa genes, thereby further increasing the toxicity of BtX023 to Helicoverpa armigera and P. xylostella by enhancing energy synthesis, the glyoxylate cycle, and branched-chain amino acids synthesis, but not poly-ß-hydroxy butyrate synthesis.

Construction of a Conditionally Asporogenous Bacillus thuringiensis Recombinant Strain Overproducing Cry Protein by Deletion of the leuB Gene.

One of the common shortcomings with Bacillus thuringiensis (Bt) biopesticides in field application is their instability under UV irradiation. In Bt, the leuB gene encodes the 3-isopropylmalate dehydrogenase. In addition to its role in leucine biosynthesis, LeuB would be likely recruited to catalyze the dehydrogenation of malate in the final step of tricarboxylic acid cycle during sporulation. In this study, we constructed a Bt recombinant strain in which the gene leuB was deleted by using the markerless gene deletion system. The ΔleuB mutant strain showed a conditionally asporogenous phenotype while overproducing insecticidal crystal proteins and retaining its insecticidal activity well in both fermentation and LB media. Furthermore, the metabolic regulation mechanisms of LeuB was elucidated by iTRAQ-based quantitative proteomics approach. Evidences from proteomics data suggested that the inhibited supply of pyruvate (carbon source) was an important factor related to the conditionally asporogenous feature of the mutant. Consistently, the mutant regained its ability to sporulate in LB medium by adding 1% glucose or 1% sodium pyruvate. Taken together, our study demonstrated that deletion of the leuB gene resulted in delayed or completely blocked mother cell lysis, allowing the crystals encapsulated within cells, which makes this recombinant strain a good candidate for developing Bt preparations with better UV-stability.

Effects of SpoIVA on the formation of spores and crystal protein in Bacillus thuringiensis.

In addition to forming spores, Bacillus thuringiensis (Bt) 4.0718 can produce toxins, insecticidal crystal protein (ICP) and vegetative insecticidal protein (Vip). The Bt spoIVA was successfully knocked out by gene recombination and was shown to inhibit sporulation. The mutant strain also exhibited significantly decreased growth and crystal formation, which inhibited spore formation and partially reduced the rate of crystal synthesis. The 50 % lethal concentrations (LC50) values of Bt 4.0718, replacement, complementation and multi-copy mutant strains against the fourth larval stage of H. armigera was determined as 5.422, 6.776, 6.223 and 5.018 µg/mL, respectively. A total of 1814 proteins were identified through isobaric tags for relative and absolute protein (iTRAQ), with 41 and 54 up and downregulated proteins observed. Gene ontology enrichment analysis showed that differentially expressed proteins were primarily involved in the biological process and molecular function. Quantitative real-time PCR analysis confirmed that 9 differential expressed genes exhibited a positive correlation between changes at transcriptional and translational levels. The results of this study provide a basis for further studies of the metabolic regulatory network of spores and crystal protein formation. Moreover, they can be used to ecologically safe insecticide of farmland production because the constructed Bt spoIVA mutants did not produce spores.Provides new ideas for the targeted improvement and application of environmentally friendly spore-free strains.

Circumnutation and Growth of Inflorescence Stems of Arabidopsis thaliana in Response to Microgravity under Different Photoperiod Conditions.

Circumnutation is a periodic growth movement, which is an important physiological mechanism of plants to adapt to their growth environments. Gravity and photoperiod are two key environmental factors in regulating the circumnutation of plants, but the coordination mechanism between them is still unknown. In this study, the circumnutation of Arabidopsis thaliana inflorescence stems was investigated on board the Chinese recoverable satellite SJ-10 and the Chinese spacelab TG-2. Plants were cultivated in a special plant culture chamber under two photoperiod conditions [a long-day (LD) light: dark cycle of 16:8 h, and a short-day (SD) light: dark cycle of 8:16 h]. The plant growth and movements were followed by two charge-coupled device (CCD) cameras. The parameter revealed a daily (24 h) modulation on both TG-2 and SJ-10, under both the LD and the SD conditions. The inhibition of circumnutation was more apparent by microgravity under the SD in comparison with that under the LD condition, suggesting the synergistic effects of the combined microgravity and photoperiod on the circumnutation in space. In addition, an infradian rhythm (ca. 21 days long) on the TG-2 was also observed.