Unveiling the microbiota of sauce-flavor Daqu and its relationships with flavors and color during maturation.

This study investigated the microbial community in three-color sauce-flavor Daqu (black, yellow, and white) throughout their maturation processes, together with their physicochemical factors, culturable microbes, flavor components, and fermenting vitalities. Results from high-throughput sequencing revealed distinct microbial diversity, with more pronounced variations in bacterial community than in fungal community. Firmicutes and Ascomycota emerged as the most dominant bacterial and fungal phyla, respectively, during maturation. Genus-level analysis identified Kroppenstedia, Virgibacillus, and Bacillus as dominant bacteria in black Daqu, yellow Daqu, and white Daqu, severally, while Thermoascus was shared as the core dominant fungi for these Daqu. Physicochemical factors, particularly acidity, were found to exert a significant impact on microbial community. Kroppenstedtia was the key bacteria influencing the color formation of these Daqu. Furthermore, correlations between dominant microbes and flavor compounds highlighted their role in Daqu quality. Molds (Aspergillus, Rhizomucor, and Rhizopus), excepting Bacillus, played a crucial role in the formation of pyrazine compounds. Consequently, this study offers innovative insights into the microbial perspectives on color and pyrazine formation, establishing a groundwork for future mechanized Daqu production and quality control of sauce-flavor baijiu.

Proteomic and metabolomic analysis of ageing beef exudate to determine that iron metabolism enhances muscle protein and lipid oxidation.

The study aimed to assess differences in proteomic and metabolite profiles in ageing (1, 2, 4, and 6 days at 4 °C) beef exudates and determine their relationship with beef muscle iron metabolism and oxidation. Proteomic and metabolomic analyses identified 877 metabolites and 1957 proteins. The joint analysis identified 24 differential metabolites (DMs) and 56 differentially expressed proteins (DEPs) involved in 15 shared pathways. Ferroptosis was identified as the only iron metabolic pathway, and 4 DMs (l-glutamic acid, arachidonic acid, glutathione and gamma-glutamylcysteine) and 5 DEPs (ferritin, phospholipid hydroperoxide glutathione peroxidase, heme oxygenase 1, major prion protein, and acyl-CoA synthetase long chain family member 4) were involved in iron metabolism by regulating heme and ferritin degradation, Fe2+ and Fe3+ conversion, arachidonic acid oxidation and inactivation of glutathione peroxidase (GPX) 4, leading to increased levels of free iron, ROS, protein and lipid oxidation (P < 0.05). Overall, abnormal iron metabolism during ageing induced oxidative stress in muscle tissue.

Dominant Contribution of NO3 Radical to NO3- Formation during Heavy Haze Episodes: Insights from High-Time Resolution of Dual Isotopes Δ17O and δ18O.

δ18O is widely used to track nitrate (NO3-) formation but overlooks NO3 radical reactions with hydrocarbons (HCs), particularly in heavily emitting hazes. This study introduces high-time resolution Δ17O-NO3- as a powerful tool to quantify NO3- formation during five hazes in three cities. Results show significant differences between Δ17O-NO3- and δ18O-NO3- in identifying NO3- formation. δ18O-NO3- results suggested N2O5 hydrolysis (62.0-88.4%) as the major pathway of NO3- formation, while Δ17O-NO3- shows the NO3- formation contributions of NO2 + OH (17.7-66.3%), NO3 + HC (10.8-49.6%), and N2O5 hydrolysis (22.9-33.3%), revealing significant NO3 + HC contribution (41.7-56%) under severe pollution. Furthermore, NO3- formation varies with temperatures, NOx oxidation rate (NOR), and pollution levels. Higher NO2 + OH contribution and lower NO3 + HC contribution were observed at higher temperatures, except for low NOR haze where higher NO2 + OH contributions were observed at low temperatures (T ← 10 °C). This emphasizes the significance of NO2 + OH in emission-dominated haze. Contributions of NO2 + OH and NO3 + HC relate to NOR as positive (fP1 = 3.0*NOR2 - 2.4*NOR + 0.8) and negative (fP2 = -2.3*NOR2 + 1.8*NOR) quadratic functions, respectively, with min/max values at NOR = 0.4. At mild pollution, NO2 + OH (58.1 ± 22.2%) dominated NO3- formation, shifting to NO3 + HC (35.5 ± 16.3%) during severe pollution. Additionally, high-time resolution Δ17O-NO3- reveals that morning-evening rush hours and high temperatures at noon promote the contributions of NO3 + HC and NO2 + OH, respectively. Our results suggested that the differences in the NO3- pathway are attributed to temperatures, NOR, and pollution levels. Furthermore, high-time resolution Δ17O-NO3- is vital for quantifying NO3 + HC contribution during severe hazes.

A therapy that modulates T lymphocyte subsets in patients infected with Epstein-Barr virus: Ganciclovir combined with interferon atomization inhalation.

To investigate the effect of ganciclovir combined with interferon atomization inhalation on T lymphocyte subsets in patients with Epstein-Barr virus (EBV) infection and its efficacy. Fifty patients with EBV infection who received ganciclovir combined with interferon atomization inhalation were selected as the observation group, and 50 healthy people were selected as the control group. The changes of T lymphocyte subsets in peripheral blood were detected by flow cytometry before treatment and at the 1st, 2nd, 3rd and 4th cycle after treatment. Before treatment, the CD3+, CD4+, CD4+/CD8+ indexes of the patients were significantly lower than those of the control group (P < .05), and the CD8+ level was significantly increased (P < .05). After one cycle of treatment, there was no significant difference in the changes of T lymphocyte subsets compared with those before treatment. After 2 and 3 cycles of treatment, CD3+, CD4+, CD4+/CD8+ values were higher than those before treatment (P > .05), and CD8+ index was lower than that before treatment (P < .05). After the 4th cycle of treatment, CD3+, CD4+, CD4+/CD8+ values were significantly improved (P < .05), and CD8+ index was significantly decreased (P < .05). Ganciclovir combined with interferon atomization inhalation can regulate the changes of T lymphocyte subsets in patients with EBV infection, improve the patient's condition, and has no obvious adverse reactions. Monitoring the changes of T lymphocyte subsets during treatment is more meaningful to predict the therapeutic effect of patients with EB virus infection.

[The Role and Mechanism of MiR-451 in Multidrug Resistance of Leukemia Cell Line K562/A02].

OBJECTIVE: To detect the differential expressions of miR-451, ABCB1 and ABCC2 in drug-sensitive leukemia cell line K562 and drug-resistant cell line K562/A02, and explore the regulatory relationship between miR-451 and the expressions of ABCB1 and ABCC2 , and the mechanism of miR-451 involved in drug resistance in leukemia. METHODS: CCK-8 assay was used to detect the drug resistance of K562/A02 and K562 cells. Quantitative Real-time PCR (qRT-PCR) was used to verify the differential expressions of miR-451 in K562 and K562/A02 cells. MiR-451 mimic and negative control (miR-NC), miR-451 inhibitor and negative control (miR-inNC) were transfected into K562 and K562/A02 cells respectively, then qRT-PCR and Western blot were used to detect the expression levels of mRNA and protein of ABCB1 and ABCC2 in K562 and K562/A02 cells and the transfected groups. RESULTS: The drug resistance of K562/A02 cells to adriamycin was 177 times higher than that of its parent cell line K562. Compared with K562 cells, the expression of miR-451 in K562/A02 cells was significantly higher (P <0.001), and the mRNA and protein expression levels of ABCB1 and ABCC2 in K562/A02 cells were significantly higher than those in K562 cells (P <0.001). After transfected with miR-451 inhibitor, the expression of miR-451 was significantly down-regulated in K562/A02 cells (P <0.001), the sensitivity to chemotherapy drugs was significantly enhanced (P <0.05), and the mRNA and protein expressions of ABCB1 and ABCC2 were significantly decreased (P <0.01). After transfected with miR-451 mimic, the expression of miR-451 was significantly upregulated in K562 cells (P <0.001), and the mRNA and protein expressions of ABCB1 and ABCC2 were significantly increased (P <0.01). CONCLUSION: There are significant differences in the expressions of miR-451, ABCB1 and ABCC2 between the drug-sensitive leukemia cell line K562 and drug-resistant cell line K562/A02, which suggests that miR-451 may affect the drug resistance of leukemia cells by regulating the expression of ABCB1 and ABCC2.

Whole genome resequencing reveal patterns of genetic variation within Colletotrichum acutatum species complex from rubber trees in China.

The Colletotrichum acutatum species complex possesses a diverse number of important traits, such as a wide host range and host preference, different modes of reproduction, and different strategies of host infection. Research using comparative genomics has attempted to find correlations between these traits. Here, we used multi-locus techniques and gene genealogical concordance analysis to investigate the phylogenetic relationships and taxonomic status of the Colletotrichum acutatum species complex using field isolates obtained from rubber trees. The results revealed that the dominant species was C. australisinense, followed by C. bannaense, while strain YNJH17109 was identified as C. laticiphilum. The taxonomic status of strains YNLC510 and YNLC511 was undetermined. Using whole-genome single nucleotide polymorphism data to analyze population structure, 18 strains of C. australisinense were subsequently divided into four populations, one of which was derived from an admixture of two populations. In addition, the strains LD1687, GD1628, and YNLC516, did not belong to any populations, and were considered to be admixtures of two or more populations. A split decomposition network analysis also provided evidence for genetic recombination within Colletotrichum acutatum species complex from rubber trees in China. Overall, a weak phylogeographic sub-structure was observed. Analysis also revealed significant differences in morphological characters and levels of virulence between populations.

Emission characteristics and influencing mechanisms of PAHs and EC from the combustion of three components (cellulose, hemicellulose, lignin) of biomasses.

Biomass burning is an important source of polycyclic aromatic hydrocarbons (PAHs) and elemental carbon (EC), but the formation mechanisms are still unclear. Cellulose, hemicellulose, and lignin are the three major components of biomass. In this study, the three-components extracted from three typical biomass raw materials were used for laboratory combustion experiments, to investigate the differences in the emission factors and chemical compositions of PAHs and EC. The average emission factors of the 16 kinds of PAHs were showing as lignin (135 ± 180 mg/kg) > cellulose (97.8 ± 124 mg/kg) > hemicellulose (48.9 ± 65.2 mg/kg), and the average emission factors of EC presented in the descending order of cellulose (1.65 ± 3.02 g/kg), lignin (1.30 ± 1.04 g/kg), and hemicellulose (0.450 ± 0.480 g/kg), respectively. The proportion of naphthalene emitted from cellulose and hemicellulose combustion is higher, while fluoranthene and pyrene accounted significantly higher proportion for lignin. Moreover, the influence of ignition temperature and oxygen content on the emission characteristics of PAHs and EC were also discussed. The influence of ignition temperature on the emission of EC and PAHs is more significant compared to oxygen content, because it obviously promoted the PAHs and EC formations through resonance-stabilized hydrocarbon-radical chain reaction (RSR) pathway. However, correlation analysis combined with cluster analysis showed that the RSR-pathway probably had different effects on PAH growth for the three-components, as the indene-involved RSR-pathway were mainly related to 4-6 ring PAHs for cellulose and lignin (except fluoranthene and pyrene), but 2-4 ring PAHs for hemicellulose. We also found that the fitted results according to the proportion of three-components were significantly higher than the measured values of raw materials for indene, medium-molecular-weight PAHs, and soot-EC. These results presented the different formation pathways for medium-molecular-weight PAHs and the two EC components emitted by biomass combustion, which are worthy of further studies in exploring the generation mechanisms of PAHs and EC.

Expression of CD123 Related Long Non-coding RNA in Acute Myeloid Leukemia Bone Marrow Mononuclear Cells and Its Clinical Significance.

Acute myelogenous leukemia (AML) is a very common hematopoietic malignancy. Hematopoietic stem cell transplantation can improve the therapeutic effect of AML, but the 5-year survival rate is very low. CD123 imbalance, abnormal gene expression, and epigenetics play an important role in the pathogenesis of AML. This research was to explore the differential expression of CD123-related long non-coding RNA (lncRNA) in AML bone marrow mononuclear cells and provide a theoretical basis for targeted therapy of AML. High-throughput sequencing was performed to screen differentially expressed lncRNA in bone marrow mononuclear immunophenotypes of CD123+ and CD123- from patients with primary AML, and real-time quantitative PCR was adopted for screening and validation. There were 933 differentially expressed lncRNAs in the CD123+ group and the CD123- group, 407 lncRNAs were up-regulated and 463 lncRNAs were down-regulated in the CD123+ group. 14 lncRNAs with more than 2 times of difference were screened for identification, and it was found that compared with CD123- group, there was no substantial difference in the expression of JHDM1D-AS1, LINC01355, CASC15, FAM13A-AS1, HSPC324, LOC339803, LINC00877, and MAG12-AS3 in CD123+ group (P>0.05). The expressions of LOC101929698, BaALC-AS2, BOLA3-AS1, and FBX19-AS1 were considerably up-regulated (P<0.05), while the expressions of LOC100132249 and LINC02085 were considerably down-regulated (P<0.05). In summary, differentially expressed lncRNAs in bone marrow samples of CD123+ and CD123- group of newly diagnosed AML patients may be involved in the process of AML and seriously affect the prognosis of patients.

Emission Characteristics and Formation Mechanism of Carbonyl Compounds from Residential Solid Fuel Combustion Based on Real-World Measurements and Tube-Furnace Experiments.

This study updated carbonyl compound (CC) emission factors (EFs) and composition for residential solid fuel combustion based on real-world measurements of 124 fuel/stove combinations in China and explored the CC formation mechanism using tube-furnace experiments with 19 fuels and low/high temperatures to explain the impact of fuel and stove on CC emission characteristics. The average EFCC values for straw, wood, and coal were 1.94 ± 1.57, 1.50 ± 0.88, and 0.40 ± 0.54 g/kg, respectively. Formaldehyde and acetaldehyde were the most abundant species, accounting for 40-60% of CCs, followed by acetone (∼20%), aromatic aldehydes (∼10%), and unsaturated aldehydes (∼5%). Different from formaldehyde and acetaldehyde, other species showed significant variation among fuel types. All these characteristics could be explained by the difference in the volatile content and chemical structure of fuel, such as aromatic in coal versus lignin in biomass. The improvement in stove technology reduced CC emissions by 30.4-69.7% (mainly formaldehyde and acetaldehyde) among fuels but increased the proportion of aromatic aldehydes by 24.3-89.4%. Various CC species showed different formation mechanisms related to fuel property and burning temperature. The volatile matter derived from thermal pyrolysis of fuel polymers determined CC composition, while higher temperature preferentially degraded formaldehyde and acetaldehyde but promoted the formation of acetone and aromatic aldehydes. This study not only revealed emission characteristic of CCs from RSFC but also contributed to the improvement of clean combustion technology.

Rituximab combined with GDP regimen can effectively improve the efficacy and immune function of non-Hodgkin lymphoma.

OBJECTIVE: To evaluate the efficacy of rituximab combined with GDP regimen (gemcitabine + cisplatin + dexamethasone) in the treatment of non-Hodgkin lymphoma and its impact on the immune function of patients. METHODS: Clinical data of 88 patients with non-Hodgkin lymphoma (NHL) treated in Affiliated Hospital of Yan'an University from February 2017 to February 2019 were analyzed retrospectively. Among them, 40 patients treated with the second-line regimen (gemcitabine + cisplatin + dexamethasone) were served as the control group, and 48 patients received additional rituximab were as the observation group. The therapeutic effect, incidence of adverse reactions, levels of complement (C3, C4) and immunoglobulin [immunoglobulin (Ig) G, IgM, IgA] before and after treatment were compared between the two groups. Cox regression analysis was used to analyze the prognostic factors of patients. RESULTS: The total response rate of patients in observation group was higher than that in the control group (P<0.05). There was no significant difference in the incidence of adverse reactions (hair loss, nausea and vomiting, thrombocytopenia, anemia and bone marrow suppression) between the two groups (P>0.05). After treatment, the levels of C3 and C4 in both groups were lower than those before treatment, and the decrease in observation group were more evident than that in control group (P<0.05). No notable fluctuation was observed in the levels of IgG, IgM and IgA in both groups between before and after treatment (P>0.05). Cox regression analysis found that Ann Arbor stage and pretreatment disease status were the factors affecting the prognosis of patients. CONCLUSION: Rituximab combined with GDP regimen has a significant effect on the treatment of non-Hodgkin lymphoma, and Ann Arbor stage and pretreatment disease state are prognostic factors for patients with NHL.

Characterization and necrosis-inducing activity of necrosis- and ethylene-inducing peptide 1-like proteins from Colletotrichum australisinense, the causative agent of rubber tree anthracnose.

Colletotrichum australisinense, a member of the Colletotrichum acutatum species complex, is an important pathogen causing rubber tree anthracnose. Genome-wide comparative analysis showed this species complex contains more genes encoding necrosis- and ethylene-inducing peptide 1-like proteins (NLPs) than other Colletotrichum species complexes, but little is known about their necrosis-inducing roles in host. The aim of this study was to analyze NLPs number and type in C. australisinense, and characterize their necrosis-inducing activity in host or non-host. According to phylogenetic relationship, conserved the cysteine residues and the heptapeptide motif (GHRHDWE), 11 NLPs were identified and classified into three types. Five of the eleven NLPs were evaluated for necrosis-inducing activity. CaNLP4 (type 1) could not induce necrosis in host or non-host plants. By contrast, both CaNLP5 and CaNLP9 (type 1) induced necrosis in host and non-host plants, and necrosis-inducing activity was strongest for CaNLP9. CaNLP10 (type 2) and CaNLP11 (type 3) induced necrosis in host but not non-host plants. Substitution of key amino acid residues essential for necrosis induction activity led to loss of CaNLP4 activity. Structural characterization of CaNLP5 and CaNLP9 may explain differences in necrosis-inducing activity. We evaluated the expression of genes coding CaNLP by reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR (qRT-PCR) at different time-points after pathogen infection. It was found that genes encoding CaNLPs with different activities exhibited significantly different expression patterns. The results demonstrate that CaNLPs are functionally and spatially distinct, and may play different but important roles in C. australisinense pathogenesis.

Emission Characteristics and Formation Pathways of Intermediate Volatile Organic Compounds from Ocean-Going Vessels: Comparison of Engine Conditions and Fuel Types.

The lack of emission data for ocean-going vessels (OGVs) and the recent fuel switching make it urgent to enhance the onboard measurement of ship emissions, especially for intermediate volatile organic compounds (IVOCs). This study focused on the IVOC emission characteristics and formation pathways of three OGVs under various engine conditions (power and load) and fuel oils (heavy fuel oil (HFO) versus marine gas oil (MGO)). The results showed that the (1) IVOC emission factors (EFIVOC) of the three OGVs increased with engine power and were higher for MGO (1494.4 ± 421.7 mg/kg) than HFO (1830.5 ± 534.5 mg/kg) and engine load is an important parameter. (2) Engine load and oil type affect the composition and volatility distribution of IVOCs. The proportion of polycyclic aromatic hydrocarbons in IVOCs increased with a higher load, and using MGO shifted IVOC components to a higher volatility in contrast to HFO. (3) The compositions of IVOCs were more like those in fuel oils under low loads than under high loads, indicating that different formation pathways of IVOCs exist for different engine loads. (4) A higher EFIVOC was observed nearshore than in open sea owing to the lower and transient engine load, which indicates the necessity of paying attention to the IVOC emissions for ships.

Existence and Formation Pathways of High- and Low-Maturity Elemental Carbon from Solid Fuel Combustion by a Time-Resolved Study.

Elemental carbon (EC) from various sources contains different sub-fractions with different properties; however, this variability poses several challenges for the accurate assessment of EC emission inventory. EC is defined using thermo-optical analysis (TOA), and its different fractions have different maturation and formation pathways. High- and low-maturity ECs have similar detection signals to those of Soot-EC and Char-EC in TOA. The emission characteristics of Soot-EC and Char-EC were affected by fuel composition and combustion temperatures. Biomass combustion generated more Char-EC than coal combustion, resulting in lower Soot-EC to Char-EC ratios. Soot-EC emissions always increased with an increasing temperature. Char-EC emissions increased with an increasing temperature at 300-900 °C in biomass combustion and decreased in coal combustion when the temperature was >600 °C, suggesting that the two ECs have different formation pathways. Time-resolved analyses of organic carbon (OC), EC, and polycyclic aromatic hydrocarbons showed that Char-EC was preferentially generated in the ignition stage with the rapid emission of OC through direct conversion of OC, whereas Soot-EC was preferentially generated during the flaming stage through gas-phase polymerization of small molecules generated from the decomposition of OC.

Intermediate Volatile Organic Compound Emissions from Residential Solid Fuel Combustion Based on Field Measurements in Rural China.

Residential solid fuel combustion (RSFC) is a key cause of air pollution in China. In these serial studies, field measurements of RSFC from 166 rural households in eastern China were conducted to update the database of emission factors (EFs) and chemical profiles of gaseous and particulate organic pollutants, and the present study focuses on the intermediate volatile organic compounds (IVOCs), which are precursors of secondary organic aerosol (SOA). The results show that the averaged EFs of IVOCs (EFIVOC) for crop straw, fuelwood, and coal are 550.7 ± 397.9, 416.1 ± 249.5, and 361.9 ± 308.0 mg/kg, respectively, which are among the EFIVOC of gasoline vehicle, diesel vehicle, non-road machinery, and heavy fuel oil vessel, and are significantly affected by fuel, stove, and combustion efficiency. The percentages of normal alkanes (n-alkanes), branched alkanes (b-alkanes), polycyclic aromatic hydrocarbons (PAHs), and unresolved complex mixture from RSFC are 3.5 ± 1.6, 8.0 ± 3.7, 17.6 ± 6.7, and 70.9 ± 8.1%, respectively, and the compositions are featured by lower b-alkanes and higher PAHs than those of vehicle sources. The proportions of some individual n-alkanes and PAHs (such as n-C12-n-C15, naphthalene, and its alkyl substituents) can be used as indicators to differentiate RSFC from vehicle sources, while methoxyphenols can be used to distinguish biomass burning from coal combustion. Based on China's energy statistics, the total IVOC emissions from RSFC in 2014 were 175.9 Gg. These data will help to update the IVOC emission inventory and improve the estimates of SOA production in China.

Fuel Aromaticity Promotes Low-Temperature Nucleation Processes of Elemental Carbon from Biomass and Coal Combustion.

Significant elemental carbon (EC) emissions from low-temperature solid fuel combustion cannot be explained by classical mechanisms ascribing EC to higher-temperature condensation (> 850 °C). The importance of fuel composition in promoting EC nucleation was investigated by studying EC and polycyclic aromatic hydrocarbon (PAH) formation at multiple-ignition temperatures (300-900 °C) using fuels with different aromatic contents (i.e., straw, wood, and coal). Biomass and coal combustion at 300 °C can produce substantial EC containing a large amount of soot-EC, a known high-temperature condensation product, possibly because aromatics reduce EC nucleation barriers, corresponding to the increasing ratios of soot-EC to char-EC from straw to coal (1.22 to 3.61). High- to low-molecular-weight PAH ratios in biomass combustion were four times lower than those in coal combustion, resulting in different EC formation atmospheres. Specifically, 31.4% of PAHs from biomass combustion were indene, compared to only 0.24% for coal, indicating that resonance-stabilized hydrocarbon-radical chain reactions dominated EC nucleation in biomass combustion. Five- to six-membered PAH ratios were always higher than one in biomass combustion but increased significantly from 0.5 to 2 with increasing temperature in coal combustion, indicating that PAHs generated through aromatic decomposition in coal could form EC through van-der-Waals forces and phenyl addition/cyclization-based covalent bonding at low and high temperatures, respectively.

Virgibacillus halodenitrificans ST-1 for fermentation of shrimp paste and hydrolysates of its protease.

The nutrition and flavor of shrimp paste came from hydrolyzation by enzymes that were produced by microorganisms. The salt-tolerant strain Virgibacillus halodenitrificans ST-1 isolated from shrimp paste was studied and used in the fermentation of shrimp paste. The strain and the protease produced by ST-1 were investigated. The optimum pH of the protease was 8.0, and the reaction temperature was 30°C. The protease showed high activity in the range of pH (5.0-11.0) and NaCl concentration (1%-15%). Divalent cations such as Ba2+, Ca2+, Mg2+, Mn2+, and Si2+ could enhance the protease activity. Residual activity of protease was more than 90% when it was incubated with PMSF and H2O2. Also, the enzyme retained more than 90% of initial activity after it was incubated with organic solvents. Variety of natural proteins could be substrates of the protease. By analyzing the release rate of free amino acids, it was predicted that the cleavage sites of the protease were mainly Glu, Asp, Gly, Leu, and Lys. Moreover, the hydrolysates of the protease had antioxidant activity, especially for DPPH and superoxide anion radical scavenging. The strain ST-1 and the protease both were excellent candidates for food industries.

First Report on Neopestalotiopsis aotearoa of Rubber Tree in China.

Natural rubber is an important industrial raw material and an economically important perennial in China. In recent years, A new leaf fall disease, caused by Neopestalotiopsis aotearoa Maharachch., K.D. Hyde & Crous, has occurred in Indonesia, Malaysia, Thailand, Sri Lanka, and other major rubber planting countries. In May and July of 2020, this disease was first found on 2-year-old rubber seedlings in two plantations located in Ledong and Baisha counties in Hainan Province, China. In the two plantations of approximately 32 ha, 15% of the rubber seedlings had the disease and the defoliation was more than 20%. The infected leaves turned yellow and watery, and dark brown and nearly round lesions of 1-2 mm in diameter were formed on the leaves. When the humidity was high, the center of the lesion was grey-white, and the lesions had many small black dots, black margins and surrounded by yellow halos. When the disease was severe, leaves fell off. To identify the pathogen, leaf tissues were collected from lesion margins after leaf samples were surface-sterilized in 75% ethanol, rinsed with sterile water for three times, and air dried. The leaf tissues were plated on potato dextrose agar (PDA) and incubated at 28°C for seven days. Fungal cultures with similar morphology were isolated from 90% of tested samples and two isolates (HNPeHNLD2001 and HNPeHNLD2002) were used in pathogenicity and molecular tests. Rubber leaves (clone PR107) were inoculated with conidial suspension (106 conidia/ml), and inoculated with PDA were used as the control, Each treatment had 3 leaves, and each leaf was inoculated with 3 spots and incubated at 28oC under high moisture conditions. Five days later, leaves inoculated with conidial suspension showed black leaf spots resembling the disease in the field, whereas the control leaves remained symptomless. The fungal cultures isolated from the inoculated tissues, had identical morphology compared with the initial isolates. Colonies on PDA were 55-60 mm in diameter after seven days at 28°C, with undulate edges, pale brown, thick mycelia on the surface with black, gregarious conidiomata; and the reverse side was similar in color. Black conidia were produced after eight days of culture on PDA. Conidia were fusoid, ellipsoid, straight to slightly curved, 4-septate, ranged from 18.35 to 27.12 µm (mean 22.34 µm) × 4.11 to 7.03 µm (mean 5.41 µm). The basal cells were conic with a truncate base, hyaline, rugose and thin-walled, 4.35 to 6.33 µm long (mean 4.72 µm). Three median cells were doliform, 12.53 to 18.97 µm long (mean 15.26 µm), hyaline, cylindrical to subcylindrical, thin- and smooth-walled, with 2-3 tubular apical appendages, arising from the apical crest, unbranched, filiform, 14.7 to 25.3 µm long (mean 19.94 µm). The basal appendages were singlar, tubular, unbranched, centric, 3.13 to 7.13 µm long (mean 5.48 µm). Morphological characteristics of the isolates were similar to the descriptions of N. aotearoa (Maharachchikumbura et al. 2014). The rDNA internal transcribed spacer (ITS) region, translation elongation factor 1-αgenes (TEF), and beta-tubulin (TUB2) gene were amplified using the primer pairs ITS1/ITS4, EF1-728F/EF1-986R and T1/Bt-2b (Pornsuriya et al. 2020), respectively. The sequences of these genes were deposited in GenBank (ITS Accession Nos.: MT764947 and MT764948; TUB2: MT796262 and MT796263; TEF: MT800516 and MT800517). According to the latest classification of Neoprostalotiopsis spp. (Maharachchikumbura et al. 2014) and multilocus phylogeny, isolates HNPeHNLD2001 and HNPeHNLD2002 were clustered in the same branch with N. aotearoa. Thus, the pathogen was identified as N. aotearoa, which is different from N. cubana and N. formicarum reported in Thailand (Pornsuriya et al. 2020; Thaochan et al. 2020). The Neopestalotiopsis leaf spotdisease of rubber tree (H. brasiliensis) was one of the most serious and destructive leaf diseases in major rubber planting countries in Asia. ( Tajuddin et al. 2020) The present study of leaf fall disease on rubber tree caused byN. aotearoa is the first report in China. The finding provides the basic pathogen information for further monitoring the disease and its control.

Expression, purification and characterization of a cold-adapted dextranase from marine bacteria and its ability to remove dental plaque.

Dental plaque is a high-incidence health concern, and it is caused by Streptococcus mutans. Dextranase can specifically hydrolyze ɑ-1,6-glycosidic linkages in dextran. It is commonly used in the sugar industry, in the production of plasma substitutes, and the treatment and prevention of dental plaque. In this research work, we successfully cloned and expressed a cold-adapted dextranase from marine bacteria Catenovulum sp. DP03 in Escherichia coli. The recombinant dextranase named Cadex2870 contained a 2511 bp intact open reading frame and encoded 836 amino acids. The expression condition of recombinant strain was 0.1 mM isopropylthio-galactoside (IPTG), and the reduced temperature was 16 °C. The purified enzyme activity was 16.2 U/mg. The optimal temperature and pH of Cadex2870 were 45 °C and pH 8, and it also had catalytic activity at 0 °C. The hydrolysates of Cadex2870 hydrolysis Dextran T70 are maltose, maltotetraose, maltopentose, maltoheptaose and higher molecular weight maltooligosaccharides. Interestingly, 0.5% sodium benzoate, 2% xylitol, 0.5% sodium fluoride, 5% propanediol, 5% glycerin and 2% sorbitol can enhance stability Cadex2870, which are additives in mouthwashes. Additionally, Cadex2870 reduced the formation of dental plaque and effectively degraded formed plaque. Therefore, Cadex2870 shows great promise in commercial applications.