Analysis of apocarotenoid volatiles from lettuce (Lactuca sativa) induced by insect herbivores and characterization of carotenoid cleavage dioxygenase gene.

Plant apocarotenoids have been shown to have a diverse biological role in herbivore-plant interactions. Despite their importance, little is known about herbivores' effect on apocarotenoid emissions in Lactuca sativa. In this study, we examined changes in apocarotenoid emissions in lettuce leaves after infestation by two insects, viz., Spodoptera littoralis larvae and Myzus persicae aphids. We found that ß-ionone and ß-cyclocitral showed higher concentrations than the other apocarotenoids, with a significant increase as per the intensity of infestation of both herbivore species. Furthermore, we performed functional characterization of Lactuca sativa carotenoid cleavage dioxygenase 1 (LsCCD1) genes. Three LsCCD1 genes were overexpressed in E. coli strains, and recombinant proteins were assayed for cleavage activity on an array of carotenoid substrates. The LsCCD1 protein cleaved ß-carotene at the 9,10 (9',10') positions producing ß-ionone. The transcript analysis of LsCCD1 genes revealed differential expression patterns under varying levels of herbivores' infestation, but the results were inconsistent with the pattern of ß-ionone concentrations. Our results suggest that LsCCD1 is involved in the production of ß-ionone, but other regulatory factors might be involved in its induction in response to herbivory. These results provide new insights into apocarotenoid production in response to insect herbivory in lettuce. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03511-4.

Insights into heat-induced molecular-level interactions between wheat and common buckwheat proteins.

This study investigated the heat-induced interactions between wheat and buckwheat proteins by heating wheat proteins, buckwheat albumin, globulin, and mixtures of wheat flour with buckwheat albumin/globulin at 50, 65, 80, 95, and 100 °C. The results showed that the cross-linking reactions of wheat glutenin with buckwheat albumin and globulin initiated at 80 and 95 °C, respectively. Buckwheat albumin decreased the extractability of α-gliadin by 35 % at 95 °C and 5.9 % at 100 °C. The linkage of buckwheat globulin to wheat glutelin prevented part of the wheat gliadin from linking to glutelin, resulting in the extractability of α- and γ-gliadin increased by 8.6 % and 11 % at 95 °C, respectively. The chemical forces results indicated that interactions between wheat and buckwheat proteins were primarily driven by disulfide bonds and hydrophobic interactions. This study provides a theoretical basis for better regulating the wheat-buckwheat protein network to improve the quality of buckwheat-enriched products.

Aloe-Emodin Induces Mitochondrial Dysfunction and Pyroptosis by Activation of the Caspase-9/3/Gasdermin E Axis in HeLa Cells.

Aloe-emodin (1,8-dihydroxy-3-hydroxymethyl-anthraquinone), derived from some Chinese edible medicinal herbs, exerts a potential anticancer activity on various cancer cells, making it a drug candidate for cancer therapy. Yet, the role of aloe-emodin in pyroptosis, a new type of cell death, is uncharacterized. In this study, we explored the molecular mechanisms of aloe-emodin-triggered pyroptosis. Aloe-emodin inhibited proliferation and migration and triggered caspase-dependent cell death of HeLa cells in a dose-dependent manner. Aloe-emodin caused mitochondrial dysfunction and induced pyroptosis by activating the caspase-9/3/GSDME axis. Transcriptional analysis showed extensive changes in gene expressions in cellular pathways, including MAPK, p53, and PI3K-Akt pathways when treated with aloe-emodin. This study not only identified a novel role of aloe-emodin in pyroptotic cell death, but also performed a systematical genome-wide analysis of cellular pathways responding to aloe-emodin, providing a theoretical basis for applying anthraquinone derivatives in the treatment of GSDME-expressing cancers.

Muscle-Related Effect of Whey Protein and Vitamin D3 Supplementation Provided before or after Bedtime in Males Undergoing Resistance Training.

There is increasing evidence that dietary protein intake with leucine and vitamin D is an important factor in muscle protein synthesis. This study investigated the combined effects of consuming whey protein and vitamin D3 in the evening before bedtime or in the morning after sleeping on muscle mass and strength. Healthy, untrained males (N = 42; Age = 18-24 year) were randomly assigned into three groups: before bedtime, after sleeping, and control. Subjects underwent a 6-week resistance training program in combination with supplements that provided 25 g whey protein and 4000 IU vitamin D3 for the before bedtime and after sleeping groups and a 5 g maltodextrin placebo for the control group. A significant increase in serum vitamin D was observed in both before bedtime and after sleeping groups. All groups experienced a significant gain in leg press. However, the control group did not experience significant improvements in muscle mass and associated blood hormones that were experienced by the before bedtime and after sleeping groups. No significant differences in assessed values were observed between the before bedtime and after sleeping groups. These findings suggest that the combination of whey protein and vitamin D supplements provided either before or after sleep resulted in beneficial increases in muscle mass in young males undergoing resistance training that exceeded the changes observed without these supplements.

Lactobacillus plantarum FRT4 alleviated obesity by modulating gut microbiota and liver metabolome in high-fat diet-induced obese mice.

Background: Obesity has become a global epidemic recognized by the World Health Organization. Probiotics supplementation has been shown to contribute to improve lipid metabolism. However, mechanisms of action of probiotics against obesity are still not clear. Lactobacillus plantarum FRT4, a probiotic previously isolated from a kind of local yogurt, had good acid and bile salt tolerance and lowered cholesterol in vitro. Objective: This study aimed to evaluate the effect of L. plantarum FRT4 on serum and liver lipid profile, liver metabolomics, and gut microbiota in mice fed with a high-fat diet (HFD). Design: Mice were fed with either normal diet or HFD for 16 weeks and administered 0.2 mL of 1 × 109 or 1 × 1010 CFU/mL dosage of L. plantarum FRT4 during the last 8 weeks of the diet. Cecal contents were analyzed by 16S rRNA sequencing. Hepatic gene expression and metabolites were detected by real-time quantitative polymerase chain reaction (PCR) and metabolomics, respectively. Results: L. plantarum FRT4 intervention significantly reduced the HFD-induced body weight gain, liver weight, fat weight, serum cholesterol, triglyceride, and alanine aminotransferase (ALT) levels in the liver (P < 0.05). Liver metabolomics demonstrated that the HFD increased choline, glycerophosphocholine, and phosphorylcholine involved in the glycerophospholipid metabolism pathway. All these changes were reversed by FRT4 treatment, bringing the levels close to those in the control group. Further mechanisms showed that FRT4 favorably regulated gut barrier function and pro-inflammatory biomediators. Furthermore, FRT4 intervention altered the gut microbiota profiles and increased microbial diversity. The relative abundances of Bacteroides, Parabateroides, Anaerotruncus, Alistipes, Intestinimonas, Butyicicoccus, and Butyricimonas were significantly upregulated. Finally, Spearman's correlation analysis revealed that several specific genera were strongly correlated with glycerophospholipid metabolites (P < 0.05). Conclusions: These findings suggested that L. plantarum FRT4 had beneficial effects against obesity in HFD-induced obese mice and can be used as a potential functional food for the prevention of obesity.

Upconversion Luminescence-Initiated and GSH-Responsive Self-Driven DNA Motor for Automatic Operation in Living Cells and In Vivo.

In light of the worthy design flexibility and the good signal amplification capacity, the recently developed DNA motor (especially the DNA walker)-based fluorescent biosensors can offer an admirable choice for realizing bioimaging. However, this attractive biosensing strategy not only has the disadvantage of uncontrollable initiation but also usually demands the supplement of exogenous driving forces. To handle the above obstacles, some rewarding solutions are proposed here. First, on the surface of an 808 nm near-infrared light-excited low-heat upconversion nanoparticle, a special ultraviolet upconversion luminescence-initiated three-dimensional (3D) walking behavior is performed by embedding a photocleavage linker into the sensing elements, and such light-controlled target recognition can perfectly overcome the pre-triggering of the biosensor during the biological delivery to significantly boost the sensing precision. After that, a peculiar self-driven walking pattern is constructed by employing MnO2 nanosheets as an additional nanovector to physically absorb the sensing frame, for which the reduction of the widespread glutathione in the biological medium can bring about sufficient self-supplied Mn2+ to guarantee the walking efficiency. By selecting an underlying next-generation broad-spectrum cancer biomarker (survivin messenger RNA) as the model target, we obtain that the newly formed autonomous 3D DNA motor shows a commendable sensitivity (where the limit of detection is down to 0.51 pM) and even an outstanding specificity for distinguishing single-base mismatching. Beyond this sound assay performance, our sensing approach is capable of working as a powerful imaging platform for accurately operating in various living specimens such as cells and bodies, showing a favorable diagnostic ability for cancer care.

Serum Metabolomics Analysis for Biomarkers of Lactobacillus plantarum FRT4 in High-Fat Diet-Induced Obese Mice.

Lactobacillus plantarum is considered a potential probiotic supplementation for treating obesity. However, the underlying molecular mechanism is poorly understood. Our previous study displayed that L. plantarum FRT4 alleviated obesity in mice fed a high-fat diet (HFD) through ameliorating the HFD-induced gut microbiota dysbiosis. To explore the roles of FRT4 in obesity prevention, in this study, we investigated changes in serum metabolomic phenotype by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS) and analyzed the pathway of HFD-fed Kunming female mice orally administered with FRT4 for eight weeks. Using orthogonal partial least squares discriminant analysis (OPLS-DA), metabolite patterns with significant changes were observed. 55 metabolites including phosphatidylcholine, lysophophatidylcholine, sphingomyelin, serotonin, indole-3-methyl aceta, indole-3-carbinol, indole-5,6-quino, 11,12-DHET, prostaglandin B2, leukotriene B4, and 3-hydroxybenzoic acid were identified as potential biomarkers associated with obesity, which were mainly involving in glycerophospholipid metabolism, tryptophan metabolism, and arachidonic acid metabolism. Perturbations of 14 biomarkers could be regulated by FRT4 intervention. These metabolites may serve as valuable biomarkers to understand the mechanisms by which intake of diets containing FRT4 contributes to the treatment or prevention of obesity. Thus, FRT4 can be a promising dietary supplement for the prevention of HFD-induced obesity.

[Research progress in structure and function of pectin methylesterase].

Pectin methylesterase (PME) is an important pectinase that hydrolyzes methyl esters in pectin to release methanol and reduce the degree of methylation of pectin. At present, it has broad application prospects in food processing, tea beverage, paper making and other production processes. With the in-depth study of PME, the crystal structures with different sources have been reported. Analysis of these resolved crystal structures reveals that PME belongs to the right-hand parallel ß-helix structure, and its catalytic residues are two aspartic acids and a glutamine, which play the role of general acid-base, nucleophile and stable intermediate, in the catalytic process. At the same time, the substrate specificity is analyzed to understand the recognition mechanism of the substrate and active sites. This paper systematically reviews these related aspects.

Production pectin oligosaccharides using Humicola insolens Y1-derived unusual pectate lyase.

The economical production of pectin oligosaccharides with a specific degree of polymerization and structure from agro-food waste is an industrially important process. This study identified a novel pectate lyase gene (plhy1) from the thermophilic cellulolytic fungus H. insolens Y1 and tested its ability to produce pectin oligosaccharides. The recombinant PLHY1 produced in Pichia pastoris was superior to other similar enzymes due to its high thermal and pH stability. PLHY1 demonstrated optimal enzymatic activity at 55°C and pH 10.0 in the presence of 0.4 mM Ca2+, and preferred methyl esterified substrates for digestion. High performance anion exchange chromatography-pulsed amperometric detector and ultra high performance liquid chromatography in combination with electrospray ionization tandem mass spectrometry analysis showed that galacturonic acid-oligosaccharides with a small degree of polymerization (4-6) were the major hydrolysates produced by the degradation of apple peel pectin by PLHY1. The properties of PLHY1 make it valuable for application in the agro-food industry for the production of pectin oligosaccharides.

Icaritin promotes tumor T-cell infiltration and induces antitumor immunity in mice.

Icaritin, a hydrolytic product of icariin isolated from traditional Chinese herbal medicine genus Epimedium, has many pharmacological and biological activities. Here, we show that icaritin can effectively decrease tumor burden of murine B16F10 melanoma and MC38 colorectal tumors in a T-cell dependent manner. The treatment effects are associated with increased CD8 T-cell infiltration and increased effector memory T-cell frequency. In vivo depletion of CD8 T cell using an anti-CD8 monoclonal antibody abolished the antitumor effect, which supports the critical role of CD8 T cells during icaritin treatment. By analyzing immune cells in the tumor tissue, we found reduced frequency of CD11b+ Gr1+ myeloid-derived suppression cells (MDSCs) infiltration and downregulation of PD-L1 expression on MDSCs after icaritin treatment. This was not limited to MDSCs, as icaritin also decreased the expression of PD-L1 on neutrophils. Importantly, the combination of anti-PD-1/CTLA-4 and icaritin significantly enhances antitumor ability and increases the efficacy of either treatment alone. Our findings reveal that icaritin induces antitumor immunity in a CD8 T-cell-dependent way and justify further investigation of combining immune checkpoint therapy to icaritin-based antitumor therapy.

Probing the role of cation-π interaction in the thermotolerance and catalytic performance of endo-polygalacturonases.

Understanding the dynamics of the key pectinase, polygalacturonase, and improving its thermotolerance and catalytic efficiency are of importance for the cost-competitive bioconversion of pectic materials. By combining structure analysis and molecular dynamics (MD) simulations, eight mutagenesis sites having the potential to form cation-π interactions were identified in the widely used fungal endo-polygalacturonase PG63. In comparison to the wild-type, three single mutants H58Y, T71Y and T304Y showed improved thermostability (the apparent Tms increased by 0.6-3.9 °C) and catalytic efficiency (by up to 32-fold). Chromatogram analysis of the hydrolysis products indicated that a larger amount of shorter sugars were released from the polygalacturonic acid by these three mutants than by the wild-type. MD analysis of the enzyme-substrate complexes illustrated that the mutants with introduced cation-π interaction have modified conformations of catalytic crevice, which provide an enviable environment for the catalytic process. Moreover, the lower plasticity of T3 loop 2 at the edge of the subsite tunnel appears to recruit the reducing ends of oligogalacturonide into the active site tunnel and initiates new hydrolysis reactions. This study demonstrates the importance of cation-π interaction in protein conformation and provides a realistic strategy to enhance the thermotolerance and catalytic performance of endo-polygalacturonases.

Probiotic (Enterococcus faecium) induced responses of the hepatic proteome improves metabolic efficiency of broiler chickens (Gallus gallus).

BACKGROUND: The liver plays important roles in nutrient metabolism, detoxification and immunity. Enterococcus faecium (E. faecium) is a probiotic that has been shown to have positive effects on broiler production. However, its molecular effects on liver metabolism have not been characterized. This study aims to further identify the biological roles of E. faecium by characterizing the hepatic proteomic changes of broilers (Gallus gallus) fed E. faecium using two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) and mass spectrometry (MS). RESULTS: Thirty-three proteins (50 protein spots) involved in nutrient metabolism, immunity and the antioxidant system were shown to be differentially expressed in the liver of broilers fed E. faecium than from birds not fed the probiotic. The biological processes of sulphur amino acids, vitamin and cellular hormone metabolism, sulphur compound biosynthesis and protein tetramerization were enhanced in the liver of broilers fed E. faecium. However, proteins involved in calcium ion flux, cell redox homeostasis and platelet activation related to hepatic immune responses were down-regulated in broilers fed E. faecium. These results indicate that the supplementation of poultry feed with E. faecium may alter the partitioning of nutrients and promote optimal nutrient utilization. CONCLUSIONS: This study assists in unraveling the molecular effects of the dietary probiotic, E. faecium, in the liver of broiler chickens. It shows that the probiotic improves the metabolism of nutrients and decreases inflammatory responses. Our findings extend previous knowledge of the mechanism of dietary probiotic action and provide new findings for research and future probiotic development.

[Effect of Montmorillonite on Fractions and Availability of Phosphorus in Soils Applied with Organic Fertilizer].

An incubation experiment was carried out to study the effect of montmorillonite on pH, the fractions and availability of phosphorus in a sandy yellow soil (SY) and a mineral flavicant soil (MF) applied with biogas residues of swine manure (BSR). The rates of montmorillonite applied were 0%, 5%, 7.5% and 10%, respectively. The results indicated that soil pH was not affected by BSR, but was significantly increased by montmorillonite. In both soils, application of BSR caused an increase in the content and proportion of Al-P during the incubation, and an increase in the content but a slight decrease in the proportion of Fe-P in later period of incubation. The concentration of O-P was not markedly changed, but its proportion was decreased by applying BSR in both soils. The changes of Ca-P varied with soils. BSR had no significant influence on Ca-P content, but decreased the proportion of Ca-P in SY, meanwhile, it increased both Ca-P content and proportion in MF. Addition of montmorillonite in soils applied with BSR decreased the contents and proportions of Al-P and O-P, and the proportion of Fe-P did not change the contents of Fe-P, but increased the content and proportion of Ca-P in SY. In MF, montmorillonite could decrease the contents and proportions of Fe-P and O-P, and the proportion of Al-P did not affect the content of Al-P, but increased the contents and proportions of Ca-P greatly. The contents of available phosphorus (A-P) were enhanced by 17.9%-38.0% and 17.1%-33.7% in SY and MF respectively, and the phosphorus activation coefficient (PVC) was significantly improved in both soils by applying BSR. The contents of A-P were reduced by 8.8%-35.5% and 1.1%-11.6% in SY and MF respectively. Correlation analysis showed that montmorillonite reduced the availability of phosphorus through increasing pH and contents of Ca-P, decreasing the content of Al-P in SY, and through increasing pH and decreasing content of Fe-P in MF. The availability of phosphorus in SY was influenced more obviously than that in MF by montmorillonite.

Biochemical characterization of three distinct polygalacturonases from Neosartorya fischeri P1.

Polygalacturonase is one of the most important industrial pectinases. To enrich the genetic resources and develop new enzyme candidates, three polygalacturonase genes (Nfpg I-III) of glycosyl hydrolase family 28 were cloned from Neosartorya fischeri P1 and functionally expressed in Pichia pastoris. The purified recombinant proteins exhibited some distinguished properties. In comparison with other counterparts, NfPG I showed the highest specific activity (40, 123 U/mg), NfPG II had the highest temperature optimum (65 °C), and the pH optimum of NfPG III was the lowest (3.5). The orders of their thermostability and resistance to chemicals tested were NfPG II>NfPG III>NfPG I and NfPG II>NfPG I>NfPG III, respectively. Combinations of these enzymes showed better performance than individuals in the processing and clarification of apple and strawberry juice. These results suggest that N. fischeri polygalacturonases have great application potentials in the food industry for juice production.

[Characterization of phosphorus forms in different organic materials].

The existing forms of phosphorus in seven organic waste materials including biogas residues of swine manure (ZZ), biogas residues of cattle manure (NZ), compost of cattle manure and corn straw (NJD), compost of sewage sludge (WD) and compost of rural daily garbage (NSLD) were characterized according to phosphorus fractionation procedures developed by DOU et al. The result showed that there was a great difference in the total phosphorus (TP) and the total phosphorus of various forms (P(t)) among different organic materials. ZZ had the highest content of TP with the value of 23.59 g x kg(-1); while NZ had the lowest TP content with the value of 3.61 g x kg(-1). The contents and proportions of phosphorus fractions in ZZ, NZ, NJD and WD followed the order of HCl-P > Residues-P > NaHCO3-P > NaOH-P > H2O-P, while followed the order of HC1-P > Residues-P > H2O-P > NaHCO3-P > NaOH-P in the three NSLDs. The proportion of HCl-P in the total fractionated phosphorus (P(tt)) in seven organic materials ranged from 47.75% to 84.96%, which indicated that most of P in organic materials existed in the forms that were easier to be extracted by strong extracting agents like HCl, which was difficult to be absorbed by plants. The inorganic phosphorus accounted for 79.72% -94.76% of the total phosphorus in the organic materials. Of all the phosphorus forms, the NaHCO3-P had the highest inorganic phosphorus fractions, but the inorganic phosphorus was mainly distributed in HCl-P. The organic phosphorus was mainly distributed in HCl-P and Residues-P. In addition, the higher proportions of inorganic phosphorus in NJD than those of NZ demonstrated that the composting process was benefit for the mineralization of organic phosphorus in organic materials and thus improving its availability.

Molecular characterization of a thermophilic endo-polygalacturonase from Thielavia arenaria XZ7 with high catalytic efficiency and application potential in the food and feed industries.

Thermophilic endo-polygalacturonases with high catalytic efficiency are of great interest in the food and feed industries. This study identified an endo-polygalacturonase gene (pg7fn) of glycoside hydrolase family 28 in the thermophilic fungus Thielavia arenaria XZ7. Recombinant PG7fn produced in Pichia pastoris is distinguished from other enzyme counterparts by its high functional temperature (60 °C) and specific activity (34382 ± 351 U/mg toward polygalacturonic acid). The enzyme exhibited good pH stability (pH 3.0-8.0) and resistance to pepsin and trypsin digestion and had a significant effect on disaggregation of soybean meal. Addition of 1 U/g PG7fn increased the pectin bioavailability by 19.33%. The excellent properties described above make PG7fn valuable for applications in the food and feed industries. Furthermore, a comparative study showed that N-glycosylation improved the thermostability and catalytic efficiency of PG7fn.

A novel low-temperature-active pectin methylesterase from Penicillium chrysogenum F46 with high efficiency in fruit firming.

A pectin methylesterase gene (pe8F46) was cloned from Penicillium chrysogenum F46 and successfully expressed in Pichia pastoris. The full-length cDNA consists of 969 bp and encodes a 322-residue polypeptide with the calculated molecular weight of 34.1 kDa. Deduced PE8F46 belongs to family 8 of carbohydrate esterases and shares 54% identity with a functionally characterised counterpart from Myceliophthora thermophile. Purified recombinant PE8F46 showed the optimal activity at pH 5.0 and 40°C, and remained 52% maximum activity even at 10°C. An orthogonal experiment was employed to determine the best conditions for firming pineapple dices. After incubation with 0.75% (w/v) PE8F46 and 0.4% calcium lactate (w/v) for 20 min, the firmness of pineapple dices was improved by 47.6%, 13.7% higher than that of a commercial pectinase complex. These results suggest that PE8F46 has application potential in the food industry.

A novel bifunctional pectinase from Penicillium oxalicum SX6 with separate pectin methylesterase and polygalacturonase catalytic domains.

A multimodular pectinase of glycoside hydrolase family 28, S6A, was identified in Penicillium oxalicum SX6 that consists of an N-terminal catalytic domain of pectin methylesterase, a Thr/Ser-rich linker region, and a C-terminal catalytic domain of polygalacturonase. Recombinant S6A and its two derivatives, S6PE (the catalytic domain of pectin methylesterase) and S6PG (the catalytic domain of polygalacturonase), were produced in Pichia pastoris. S6A was a bifunctional protein and had both pectin methylesterase and polygalacturonase activities. Three enzymes showed similar biochemical properties, such as optimal pH and temperature (pH 5.0 and 50 °C) and excellent stability at pH 3.5-6.0 and 40 °C. Most metal ions tested (Na(+), K(+), Ca(2+), Li(+), Co(2+), Cr(3+), Ni(2+), Cu(2+), Mn(2+),Mg(2+), Fe(3+), Zn(2+), and Pb(2+)) enhanced the pectin methylesterase activities of S6PE and S6A, but had little or inhibitory effects on the polygalacturonase activities of S6A and S6PG. In comparison with most fungal pectin methylesterases, S6A had higher specific activity (271.1 U/mg) towards 70 % DM citrus pectin. When S6PE and S6PG were combined at the activity ratio of 1:4, the most significant synergistic effect was observed in citrus pectin degradation and degumming of sisal fiber, which is comparable with the performance of S6A (95 v.s. 100 % and 16.9 v.s. 17.2 %, respectively). To the best of our knowledge, this work represents the first report of gene cloning, heterologous expression, and biochemical characterization of a bifunctional pectinase with separate catalytic domains.

Abundance and genetic diversity of microbial polygalacturonase and pectate lyase in the sheep rumen ecosystem.

BACKGROUND: Efficient degradation of pectin in the rumen is necessary for plant-based feed utilization. The objective of this study was to characterize the diversity, abundance, and functions of pectinases from microorganisms in the sheep rumen. METHODOLOGY/PRINCIPAL FINDINGS: A total of 103 unique fragments of polygalacturonase (PF00295) and pectate lyase (PF00544 and PF09492) genes were retrieved from microbial DNA in the rumen of a Small Tail Han sheep, and 66% of the sequences of these fragments had low identities (<65%) with known sequences. Phylogenetic tree building separated the PF00295, PF00544, and PF09492 sequences into five, three, and three clades, respectively. Cellulolytic and noncellulolytic Butyrivibrio, Prevotella, and Fibrobacter species were the major sources of the pectinases. The two most abundant pectate lyase genes were cloned, and their protein products, expressed in Escherichia coli, were characterized. Both enzymes probably act extracellularly as their nucleotide sequences contained signal sequences, and they had optimal activities at the ruminal physiological temperature and complementary pH-dependent activity profiles. CONCLUSION/SIGNIFICANCE: This study reveals the specificity, diversity, and abundance of pectinases in the rumen ecosystem and provides two additional ruminal pectinases for potential industrial use under physiological conditions.

A transgenic study on affecting potato tuber yield by expressing the rice sucrose transporter genes OsSUT5Z and OsSUT2M.

In many plants, sucrose transporters are essential for both sucrose exports from sources and imports into sinks, indicating a function in assimilate partitioning. To investigate whether sucrose transporters can improve the yield of starch plant, potato plants (Solanum tuberosum L. cv. Désirée) were transformed with cDNAs of the rice sucrose transporter genes OsSUT5Z and OsSUT2M under the control of a tuber-specific, class-I patatin promoter. Compared to the controls, the average fructose content of OsSUT5Z transgenic tubers significantly increased. However, the content of the sugars and starch in the OsSUT2M transgenic potato tubers showed no obvious difference. Correspondingly, the average tuber yield, average number of tubers per plant and average weight of single tuber showed no significant difference in OsSUT2M transgenic tubers with controls. In the OsSUT5Z transgenic lines, the average tuber yield per plant was 1.9-fold higher than the controls, and the average number of tubers per plant increased by more than 10 tubers on average, whereas the average weight of a single tuber did not increase significantly. These results suggested that the average number of tubers per plant showed more contribution than the average weight of a single tuber to the tuber yield per plant.