Musk deer (Moschidae), a primitive lineage within Ruminantia, are distributed across East Asia and have long been of interest in molecular phylogenetic research. The spotted forest musk deer from Huanglong (HL) Mountain in Shaanxi, China, has long been morphologically classified as Moschus moschiferus (Siberian musk deer). However, its true taxonomic status has remained uncertain due to the lack of comprehensive molecular evidence. Moreover, studies on mitochondrial genome (mitogenome) variation within Moschidae, particularly at the intraspecific level, remain limited. To date, few phylogenetic analyses of Moschidae have incorporated all available complete mitochondrial genomes from public databases. In this study, we sequenced and assembled the complete mitogenomes of two spotted musk deer individuals from Huanglong Mountain, Shaanxi, China, and identified them as M. berezovskii (forest musk deer) by phylogenetic analysis. All available complete mitochondrial genomes of Moschidae were also included in the phylogenetic reconstruction. The average complete mitogenome length of M. berezovskii distributed in HL was 16,355 bp, and comprises 13 protein-coding genes (PCGs), two ribosomal RNA genes, and 22 transfer RNA genes. Phylogenetic analysis and genetic distance indicate that the taxonomic identity of the HL Mountain population as M. berezovskii supports the monophyly of M. berezovskii and provides a robust phylogenetic framework for clarifying evolutionary relationships within the family.

gamma-aminobutyric acid (GABA), a non-protein amino acid, is a major inhibitory neurotransmitter within the human nervous system. Research on enhancing GABA production through the co-culture of yeast and lactic acid bacteria is limited, and the intricacies of their interactions and the mechanisms behind GABA enhancement are not fully elucidated. In this study, five yeast strains from Sichuan Pao Cai were co-cultured with Enterococcus faecium AB157 to increase GABA content, and it was found that combining E. faecium AB157 with Saccharomyces cerevisiae SC125 at a 4:1 ratio, along with 12 g/L MSG, at 35 degrees C, optimized GABA production to reach levels of 6.57 g/L. Within the co-culture system, there was a notable increase in both glutamate decarboxylase activity (GAD) and medium pH, while the activity of Autoinducer-2 (AI-2) remained stable. Furthermore, the cell-free supernatants (CFS) from S. cerevisiae SC125 were able to enhance GABA production by E. faecium AB157, up to 4.61 g/L. RNA sequencing revealed that the GABA biosynthetic pathways and quorum sensing (QS) in co-fermentation on E. faecium AB157 were upregulated, and the ATP synthesis and arginine deiminase pathway were downregulated compare to the pure culture of E. faecium AB157.

A co-culture strategy has been proposed for the production of desired products. However, the interaction effect on enhancing γ-aminobutyric acid (GABA) production requires further investigation. In this study, the mechanism by which Lactiplantibacillus plantarum BC112 cell-free supernatant (CFS) enhances GABA production in Enterococcus faecium AB157 was investigated, including physiological characteristics, quorum sensing (QS), and mRNA expression. The results showed CFS can significantly enhance the production of γ-aminobutyric acid (GABA) in E. faecium AB157, with a maximum increase of 4.6 times compared to the strain without CFS. CFS promoted the growth of E. faecium AB157, enhanced the activity of glutamate decarboxylase (GAD) and ATPase, and reduced the pH. The effect of CFS on QS was demonstrated through changes in Autoinducer-2 (AI-2) activity and luxS expression. Moreover, 16 amino acids were detected in CFS, with glutamic acid having the highest content. Additionally, 974 genes of E. faecium AB157 differed significantly in cultures with added CFS, which involved glycolytic, amino acid, and GABA biosynthetic pathways. Therefore, Lb. plantarum BC112 CFS enhances GABA production in E. faecium AB157 by affecting energy metabolism, the QS system, and the GABA biosynthesis system.

为探究光照强度对植物连作障碍的影响规律及机理,开展了3季不同光照强度(30、150和300μmol/m2/s)下的本生烟(Nicotiana benthamiana)连续种植试验,测量植物每季的生物量、生长指标、光合色素含量、光合作用参数及土壤有机物含量.结果表明,各光照强度下的本生烟均出现了明显的连作障碍,其中高光强下的连作障碍程度最高.通过土壤有机物分析发现,相比于未种植的初始土壤,连作土壤中的2,4-二叔丁基苯酚含量显著提升,且中高光强下的含量高于低光强.在中高光强刺激下,植物根系分泌的2,4-二叔丁基苯酚等酚酸类化合物含量显著增加并随种植季度累积于土壤,从而影响植物光合作用,抑制植物生长.

为进一步提高γ-氨基丁酸(γ-aminobutyric acid,GABA)产量,以从泡菜中筛选获得的产GABA屎肠球菌(Enterococcus faecium)AB157与植物乳杆菌(Lactobacillus plantarum)BC112为研究对象,采用高效液相色谱法对菌株共培养发酵产GABA表达能力进行评估.通过单因素试验和响应面优化共培养条件,并在最优条件下对共培养体系和单菌发酵进行谷氨酸脱羧酶酶活力分析.结果表明,当L-谷氨酸钠质量浓度为12.7 g/L、屎肠球菌AB157与植物乳杆菌BC112接种体积比例为5:3、发酵时间85 h时,共培养产GABA能力最强,达6.35 g/L,较屎肠球菌AB157单菌株发酵产GABA(1.60 g/L)提高3.9倍.在此条件下分析谷氨酸脱羧酶活力,在发酵过程中,菌株共培养体系中谷氨酸脱羧酶活力明显高于单一屎肠球菌AB157发酵酶活力.

Alpine musk deer, Moschus chrysogaster, a solitary, primitive ungulate inhabiting high elevation areas (3000–4500 m) is an endangered species facing threat of extinction globally due to excessive hunting for its musk. In this study, we determined the complete mitochondrial genome of M. chrysogaster, which was 16,354 bp in length, and revealed the same gene order and genomic organization as typical Moschidae mitochondrial DNA. Start codons in 13 protein-coding genes (PCGs) were all typical ATGs except ATA for ND2 and ND3 and ATT for ND5. Stop codons were all typical types except an incomplete stop codon T for COX3, ND2, ND3, and ND4. Secondary structures in 22 transfer RNA genes all showed typical cloverleaf except tRNA-Ser (AGY), in which the dihydrouridine arm formed a simple loop. No repeat units were found in the control region. The topology structure indicated that M. cupreus was primitive and located at the root of the Moschidae clade. Phylogenetic reconstruction placed M. chrysogaster as a distinct lineage, closely related to the branch of M. leucogaster, M. berezovskii (wild) and predicted a sister relationship with M. moschiferus, M. anhuiensis, and M. berezovskii (captive). However, we suggested that the genetic resources of M. chrysogaster_JQ608470 should be further investigated.

为拓宽产γ-氨基丁酸(γ-aminobutyric acid,GABA)微生物资源,以四川泡菜为分离源从中筛选高产GABA乳酸菌菌株,并对高产乳酸菌菌株进行生理生化、分子生物学鉴定,高产GABA发酵条件优化及其益生特性分析.结果 表明:采用高效液相色谱对菌株产GABA能力分析发现菌株AB157的GABA产量最高为1.08 g/L.生理生化和16SrRNA鉴定菌株AB157为屎肠球菌(Enterococcus faecium).通过单因素实验和响应面优化,确定屎肠球菌AB157的最佳培养条件为L-谷氨酸钠底物浓度5.2 g/L、发酵温度31℃、初始pH7、发酵时间70 h,在此条件下,屎肠球菌AB157的GABA产量达到1.60 g/L.屎肠球菌AB157的益生分析表明,其在pH2.0和3g/L的胆盐环境中的存活率分别为39％和59％,胆固醇的脱除量为18.09 μg/mL,说明该菌株具有良好的耐酸和耐胆盐特性以及较强的降胆固醇能力,可作为潜在功能性乳酸菌资源进行后续研究开发.

为拓宽高产葡聚糖的种质资源,以四川泡菜为分离源从中筛选高产葡聚糖的乳酸菌菌株,采用苯酚硫酸法对筛选菌株进行葡聚糖产量测定,并对菌株进行生理生化及16S rRNA基因和rpoB看家基因序列的同源性分析鉴定和高产葡聚糖发酵条件优化.结果 表明:菌株LM 187产葡聚糖能力较强,达到38.24 g/L,进一步鉴定为肠膜状明串珠菌肠膜亚种(Leuconostoc mesenteroides subsp.mesenteroides);采用响应面中心组合实验优化产葡聚糖发酵条件,得到最优培养条件为发酵时间35 h、发酵温度30℃、蔗糖质量分数21％和接种量3％.在此优化条件下,葡聚糖产量为56.54 g/L,较优化前提高了1.48倍.肠膜状明串珠菌肠膜亚种LM187具有高产葡聚糖的能力,可作为开发保健型泡菜的潜在优质功能性微生物资源.

One elite γ-aminobutyric acid (GABA) producing strain of Saccharomyces cerevisiae SC125 was isolated and identified from Chinese traditional paocai. S. cerevisiae SC125 and Lactobacillus plantarum BC114 were used as cooperative species to ferment mulberry (Morus alba L.) and produce a novel beverage enriched with GABA. The GABA, organic acids and volatile compounds in different fermentation stages were determined by high-performance liquid chromatography (HPLC) and headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS). It was noted that coculture changed the profiles of flavor compounds in mulberry beverage. The tartaric and succinic acid contents increased to 1.34 g/L and 0.39 g/L, respectively. Lactic, malic, citric, and oxalic acid levels ranged between 0.92 and 2.56 g/L, and ethanol and glycerol were produced at 2.66 g/L and 1.81 g/L, respectively. More volatile compounds were detected in the coculture with significantly enhanced concentrations of fruity esters including ethyl caproate, ethyl propionate, butyl acetate, ethyl butyrate, ethyl caprylate and ethyl caprate, and alcohols of phenylethyl alcohol, 1-pentanol and 2-amino-1,3-propanediol. Also, a yield of 2.42 g/L GABA was achieved in the coculture. In conclusion, S. cerevisiae SC125 and L. plantarum BC114 coculture promotes the production of flavor compounds and GABA in mulberry beverage brewing.

Objective:Lactic acid bacteria harbored plasmid was isolated from Sichuan pickles and the sequence and function of plasmid was detected.Methods:The lactic acid bacteria harbored plasmid was screened by traditional microbial separation and alkaline lysis and then the strain was identified by morphological and physiological characteristics and its 16S rDNA sequence.Plasmid was digested with single restriction enzymes and the target DNA fragments were purified and sequenced.Then the whole sequence was finally elucidated by primer design and PCR cloning.Results:A strain harbored plasmids was isolated from Sichuan pickles and identified as Weissella cibaria.A large plasmid designated as pA010 was isolated from Weissella cibaria W5 and was 11630 bp in length with GC content of 34.6％.Sequence analysis indicated that the plasmid pA010 contained four known proteins,including tyrosine recombinase,replicative protein,antoxin PemI and toxin PemK.Based on tyrosine recombinase and replication protein,pA010 may belong to a theta-type replication plasmid.In addition,PemI and PemK,an Ⅱ-TA system,may have a role in the stability of the plasmid and enhance the adaptability of bacteria to the surrounding environment.Conclusion:Weissella cibaria W5 harbored plasmid and the sequence analysis of its plasmid pA010 provided a probability for construction vectors benefited to environmental stress tolerance.

The temporal heterogeneity of prokaryotic microorganism communities in sediment of traditional freshwater cultured fish ponds in Southwest China were investigated by 16S rRNA clone libraries. The result indicated that a total of 47 archaea clones and 392 bacterial clones were recovered from three samples, among them, the bacteria Mitsuaria chitosanitabida, Prolixibacter bellariivorans and Flavihumibacter petaseus and archaea Methanoregula formicica were dominant in the cultivation 1 year (Y1) sample, the bacteria Cetobacterium somerae, Allochromatium vinosum and Rhabdochromatium marinum and archaea Methanolinea tarda were dominant in the cultivation 2 years (Y2) sample, the bacteria Cetobacterium somerae and Bacteroides fragilis and archaea Methanolinea tarda were dominant in the cultivation 4 years (Y4) sample. The Shannon–Wiener (H) diversity index and the Simpson dominancy index respectively varied from 2.23 to 2.37 and from 7.30 to 9.04 for three gene libraries. Maximum operational taxonomic unit (OTU) richness was respectively estimated with two different models SChao1 and SACE, and it revealed SChao1 15 and SACE 15 for Y1, SChao1 15.25 and SACE 15 for Y2, SChao1 12 and SACE 12 for Y4. However, the B. fragilis which was found in the Y4 sample was 99% similar to the human and animal pathogen B. fragilis. No pathogenic organisms related to humans or animals were found at any other sample. Mentionable, the more detailed information about the safety of fish pond cultivation environment should receive more attention and further studies.

以1株产γ-氨基丁酸(γ-aminobutyric acid,GABA)植物乳杆菌(Lactobacillus plantarum)BC114谷氨酸脱羧酶为研究对象,通过聚合酶链式反应(polymerase chain reaction,PCR)技术获得该酶基因,对其进行生物信息学分析,并转入大肠杆菌BL21(DE3)中实现异源表达.采用实时荧光定量PCR、聚丙烯酰胺凝胶电泳和高效液相色谱分别测定重组菌不同发酵时间点谷氨酸脱羧酶基因的表达量、蛋白表达量以及产GABA能力.结果表明:植物乳杆菌中谷氨酸脱羧酶基因大小为1410 bp,编码469个氨基酸,蛋白二级结构由α-螺旋(32.2％)、β-折叠(11.5％)和无规则卷曲(56.3％)构成,完成了该酶的三维结构同源建模;成功构建了重组谷氨酸脱羧酶大肠杆菌,谷氨酸脱羧酶基因在诱导6 h相对表达量最大,而蛋白表达量较基因在转录水平表达量有一定滞后,在8 h达最大值,此时GABA产量也达最高,为2387 mg/L.

The dietary fiber was as material which was extracted from Pleurotus eryngii residue using enzyme hydrolysis method.The application of the high-quality dietary fiber in crisp biscuit was researched.Based on the formulation of ordinary crisp biscuit,the optimum content and particle sizes of dietary fiber from Pleurotus eryngii residue in crisp biscuit were analyzed using sensory evaluation texture analysis,color value testing and flavor analysis.The results showed that the color,texture and taste were best in the dietary grain size of 120 mesh.Based on the mesh to evaluate the sensory score,it was firstly increased and then decreased with the adding of dietary fiber,especially the score of 10 percent of dietary fiber was higher than others.Totally,there were obvious difference in hardness and brittleness (p ＜ 0.05),while no significant difference in chewiness and cohesion (p ＞ 0.05).After adding dietary fiber,color value L * of biscuit decreased while a* and b* value increased.Meanwhile,it also had a positive effect on the flavor of the biscuits.Then the optimum adding amount of Pleurotus eryngii dietary fiber was 10％ with the particle size in 120 mesh,the quality of crisp biscuit was best.

In order to obtain yeasts which could produce γ-aminobutyric(GABA) and brew good performance mulberry wine,3 strains of GABA-producing yeast were screened from Sichuan pickles by paper chromatography.Then,they were identified as S.cerevisiae and C.tanzawaensis using the physiological and biochemical tests and the phylogenetic analysis.The performance of GABA-producing yeasts was assessed by high-performance liquid chromatography (HPLC) method.The results showed that the S.cerevisiae JM037 produced higher yield of GABA,and the content of GABA reached 670 mg/L.Then S.cerevisiae JM037 was further analyzed for its tolerance properties and mulberry wine brewing ability.The results showed that S.cerevisiae JM037 had good comprehensive tolerance properties and it GABA-producing capacity increased to 1145 mg/L at 144 h.Ethanol production was 38.36 g/L at 168 h,while residual sugar dropped to 3.06 g/L at 216 h.In addition,ethanol synthesis rate and the residual sugar consumption rate of S.cerevisiae JM037 was relatively slow compared with angel yeast,but S.cerevisiae JM037 has a unique capacity to produce GABA.Therefore,S.cerevisiae JM037 has potential for brewing fruit wine with GABA.

Flavors change of mulberry wine fermented by Saccharomyces cerevisiae JM037 with γ-aminobutyric acid (GABA) production capacity were analyzed during fermentation process.The changes of organic acid and volatile compounds were evaluated by high performance liquid chromatography (HPLC) and head-space solid phase micro-extraction associated with gas chromatography-mass spectrometry (HS-SPME/GC-MS),respectively.Meanwhile,the production of GABA in mulberry wine was measured.The results showed that 7 kinds of organic acids were detected,and their contents were firstly increased and then decreased.Totally 54 volatile compounds were identified in mulberry wine,including 19 esters,16 acohol,8 aldehydes,3 ketones,3 phenols,4 others,and the relative content of volatile compounds from high to low was as follows:alcohols,esters,ketones,aldehydes and phenols.Moreover,the kinds and relative content of volatile compounds were changed during fermentation process.Finally,the principal component analysis (PCA) demonstrated that aldehydes,ketones and phenols of mulberry provided the precursor substance for other flavor compounds of the fermented mulberry wine.Furthermore,the concentration of GABA in mulberry wine reached 1 180 mg/L.

Some gamma-aminobutyric acid (GABA) producing and nitrite reducing lactic acid bacteria were isolated from Sichuan paocai, a Chinese traditional fermented vegetable. Determination of GABA and sodium nitrite residue was carried out by thin layer chromatography combined with HPLC and colorimetric analysis, respectively. It was found that BC114 strain which was identified and designated as Lactobacillus plantarum BC114 possessed the highest GABA producing and nitrite degradation capacity in MRS broth with 5 g/L monosodium glutamate (MSG) and 100 mg/L sodium nitrite. Then, GABA production and nitrite degradation ability were enhanced by optimizing culture conditions, such as time, pH, temperature and MSG concentration. The greatest yield (3.45 g/L) of GABA was determined after 72 h fermentation in MRS with 20 g/L MSG at 30 degrees C and initial pH 5.5. Furthermore, nitrite degradation ability reached the maximum value of 90.17%. These results suggested that L. plantarum BC114 with GABA producing and nitrite reducing ability has the potential to be starter culture and promote the development of functional fermented foods.

The aim of the present study was to optimize the fermentation conditions of γ-aminobutyric acid (GA-BA)-producing by lactic acid bacteria from Sichuan Pickle.The performance of GABA-producing strains was evaluated by high performance liquid chromatography (HPLC) method.The results showed that strain BC114 had the higher GABA-producing ability (1.72 g/L) in MRS broth with 10 g/L initial glutamic acid after fermented at 37 ℃ for 48 h.On the basis of MRS culture medium,fermentation conditions of Lactobacillus plantarum BC114 for GABA were optimized by response surface central composite design (CCD).The results showed that the optimal fermentation medium was comprised of glucose 15 g/L,beef extract 10 g/L,peptone 10 g/L,yeast extract 5 g/L,trisodium amine 2 g/L,dipotassium phosphate 1.50 g/L,L-glutamate 17 g/L,sodium acetate 5 g/L,manganese sulfate 0.05 g/L,magnesium sulfate 0.10 g/L,and Twain-80 1 mL/L.And the optimal fermentation condition was as follows:pH 5.50,fermentation temperature 37 ℃,fermentation time 80 h and inoculum amount 4％.Under the optimized conditions,the production of GABA by L.plantarum BC114 reached 3.82 g/L,which was 2.22-fold higher than that before the optimization.

为拓展纤维素降解菌资源,以羧甲基纤维素钠(CMC-Na)为唯一碳源作初筛培养基,从浓香型白酒发酵副产物黄水中分离得到18株具有产纤维素酶能力的菌株进行纯培养.形态学、生理生化和系统发育鉴定结果显示,菌株XH01、XH04、XH05、XH18为Bacillus cereus,菌株XH34为Bacillus circulans,菌株SW01、SW05为Bacillus megaterium,菌株SW02为Bacillus endophyticus,菌株SW03、SW04为Bacillus simplex,菌株SW09、SW13为Bacillus bataviensis.菌株ZL08为Penicillium camemberti,菌株ZL13为Aspergillusfumigaus,菌株ZL04和ZL25为Penicillium chrysogenum,菌株ZL15和ZL17为Altemaria tenuissima.利用二硝基水杨酸法对菌株发酵液纤维素酶活进行研究,结果表明,菌株SW02的产酶活性较高,其羧甲基纤维素酶活为153.36 U/mL,β-葡萄糖苷酶活为126.00 U/mL,微晶纤维素酶活为17.64 U/mL,滤纸酶活为30.48 U/mL.

Oil–tea camellia see d oil was handled by two different heating methods that included the microwave and induction cooker. The variations of peroxide value,iodine value,acid value, saponification value and color values of oil–tea camellia seed oil were measured at different heating power and time in the heating process to investigate the influence of microwave heating and induction cooker on oil–tea camellia seed oil quality. And a comprehensive evaluation was given about the influence of different heating methods and different heating power on the oil–tea camellia seed oil quality. Thus,a better cooking heat way would be obtained.