冷藏过程中原料乳脂质代谢变化的靶向脂质组学分析

为探究原料乳在4 ℃冷藏过程中的脂质变化，从而指导乳及乳制品的后期加工，该研究采用超高效液相色谱-三重四级杆复合线性离子阱质谱技术分别对冷藏第0、2、3、4、6天的原料乳脂质进行绝对定性定量分析。结果表明，原料乳共检出20种脂质亚类、880种脂质分子，其中甘油三脂、磷脂酰乙醇胺、磷脂酰胆碱亚类含量最高；以P值 <0.05，变量重要性投影（variable importance projection，VIP）>1为标准共筛选出420种显著性差异脂质代谢物，以P值 <0.01，VIP >1为标准共筛选出98种极显著性差异脂质代谢物。整个冷藏过程中，极显著性差异脂质分子含量呈下降趋势，其中第3天与第4天对比组（D3 vs D4）脂质变化最为明显，冷藏3～4 d是原料乳脂质变化的关键阶段。对D3 vs D4组的极显著差异脂质进行京都基因与基因组百科全书（kyoto encyclopedia of genes and genomes，KEGG）通路分析，10种脂质分子被注释到6条代谢途径中，其中磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰肌醇、磷脂酰基丝氨酸被注释到甘油磷脂代谢及神经酰胺、鞘磷脂被注释到鞘脂代谢是该阶段的主要代谢途径。研究结果为探明原料乳冷藏过程中的脂质变化、划定脂肪水解的关键期提供理论依据，进而为后期乳制品的加工提供数据参考。     

土壤宏基因组学研究方法与进展

土壤微生物驱动着土壤中的物质循环和养分转化。在土壤学的研究中,长期将土壤作为一个黑箱系统来对待,对其中的生物组成及其参与的生化过程知之甚少。土壤中绝大部分微生物目前尚难以分离培养,因此基于传统的培养方法对于认识土壤微生物群落组成和功能有其局限性。宏基因组学直接从环境样品中提取全部微生物的DNA,或通过测序探究环境中微生物的群落结构和功能(序列驱动),或构建宏基因组文库,筛选新的基因或生物活性物质(功能驱动),克服了传统培养方法的缺陷,极大地丰富了对土壤微生物多样性及其功能的认知。本文在综述土壤宏基因组学研究基本流程的基础上,重点介绍了日益重要的第二代测序平台在土壤宏基因组学研究中的应用及其产生的海量数据的分析处理方法,并简要探讨了宏基因组学在土壤微生物生态学中的应用。最后,作者建议在国家层面上展开相关土壤宏基因组学研究,调查微生物群落及其变化,为生物资源开发、农业生产和环境保护作出应有的贡献。     

基于宏基因组学的酸性森林土壤氨氧化微生物群落特征研究

全球30％以上陆地面积是酸性土壤(pH<5.5),而酸性土壤中氨氧化微生物群落特征研究是破译其硝化过程微生物学机理的基础.尤其随着完全硝化微生物(Complete ammonia oxidizer,comammox)的发现,亟需重新认知酸性土壤中氨氧化微生物类群.以酸性马尾松林为研究对象,综合利用荧光定量PCR(qPC...     

半纤维素分解菌群HMC的微生物群落结构及功能

为明确半纤维素分解菌群HMC（hemicellulolytic microbial consortium）的微生物群落结构及其在半纤维素降解中的潜在功能;该研究测定了其在降解半纤维素主链木聚糖及玉米芯的典型理化指标,并利用16S rDNA测序技术及生物信息学手段对其微生物群落结构及功能注释进行了宏基因组学分析。结果表明,HMC在7d内能高效降解木聚糖和玉米芯,降解率达80%,木聚糖降解产生的高值产物包括还原糖、甲酸、乳酸及乙酸,最高浓度分别为1.30、0.50、1.19与1.23 mg/mL,HMC降解玉米芯时可累积挥发性脂肪酸浓度高达3.54 mg/mL。HMC是由厚壁菌门Firmicutes的多个属的微生物组成的复合微生物菌群,宏基因组分析揭示了HMC在木聚糖降解过程中的关键碳水化合物活性酶系;HMC的半纤维素降解酶系完整,包括19个木聚糖分解及下游产物代谢相关的酶;同时涵盖了乙酸、乳酸等半纤维素关键分解产物代谢相关的通路。HMC蕴含半纤维素及玉米芯高值生物转化的潜力,该研究为深入探究半纤维素在生物降解应用中的分子机制提供了重要线索,有助于推动天然半纤维素生物降解技术的发展。     

植物乳杆菌对苏尼特羊肠道菌群、血浆代谢物及肉品质的影响

为了研究植物乳杆菌对苏尼特羊肠道菌群、血浆代谢物和肉品质的影响及其作用机理,选取三月龄苏尼特羊为试验对象,随机分为2组:对照组(C组,基础日粮)和植物乳杆菌组(R组,基础日粮、活菌数为3×1010cfu/g植物乳杆菌),进行为期90 d的饲喂试验.屠宰后取其肠道内容物及背最长肌,利用高通量测序技术、代谢组学液相-色谱联...     

玉米秸秆分解过程中细菌群落组成演化特征

【目的】土壤中存在着大量的分解秸秆的微生物。研究秸秆分解过程中细菌群落组成的演化规律,对了解和调控农田微生物群体组成以促进秸秆分解具有重要意义。【方法】试验于2014年10月至2015年10月在河南省农业科学院原阳试验基地进行,将成熟期玉米秸秆(茎和叶)烘干,剪成长1~2 cm、宽0.3~1 cm的碎片,称12 g样品(相当于8 t/hm^2)装入15 cm×10 cm的尼龙网包(孔径0.04 mm)内,于10月5日冬小麦出苗后埋置在小麦垄间。分别于埋置后0、1、2、4、7、10和12个月收集秸秆包和土壤样品。测定秸秆样品干物质量和碳氮含量,选择埋置了0、2、4、7和12个月的秸秆及其土壤样品分析细菌丰度及群落组成。【结果】秸秆埋入土壤后的前2个月内分解最快,然后逐步减慢,在1、2、4、7、10和12个月后分别降解了总生物量的19.2%、32.9%、44.2%、52.2%、66.8%和73.8%。秸秆埋入土壤后,秸秆和土壤中细菌丰度均显著增加,分别于第4和7个月达到最高后开始下降。秸秆细菌的丰度指标OTUs、ACE、Chao1和多样性指标Shannon随试验时间的延长逐步增加,而Simpson指数随试验时间延长逐步降低,而土壤中这些指标在试验过程中没有显著变化。与刚埋置秸秆时相比,埋置2个月后的秸秆细菌Bacteroidetes门相对丰度明显增加,主导细菌群为Bacteroidetes和Proteobacteria门。Actinobacteria丰度在埋置2个月后明显降低,然后又随试验时间延长逐步增加。Planctomycetes、Saccharibacteria、Verrucomicrobia、Acidobacteria、Chloroflexi和Gemmatimonadetes丰度在原始秸秆中较低,埋入土壤后随试验时间延长逐步增加。Sphingobacteriia、Gammaproteobacteria、Alphaproteobacteria和Flavobacteriia主导前期细菌纲组成,而Actinobacteria、Anaerolineae和Bacilli纲丰度在后期逐步增加。秸秆分解速率主要受其碳含量影响,秸秆细菌群落组成前期与秸秆碳含量相关,后期与秸秆氮含量相关。随着试验的进展,秸秆细菌群落组成与土壤中的细菌群落组成趋同。【结论】秸秆埋入土壤后前2个月的分解速率最高,随后逐步降低。秸秆分解前期细菌群落由富营养型组分Bacteroidetes和Proteobacteria门和Sphingobacteriia、Gammaproteobacteria、Flavobacteriia和Alphaproteobacteria纲主导,随后被逐步增加的贫营养型组分Actinobacteria、Acidobacteria、Chloroflexi、Saccharibacteria门和Deltaproteobacteria、Actinobacteria纲等代替。秸秆碳氮含量变化是影响秸秆分解及其过程中细菌群落演化的主要原因。     

玉米秸秆分解过程中细菌群落组成演化特征

【目的】土壤中存在着大量的分解秸秆的微生物。研究秸秆分解过程中细菌群落组成的演化规律,对了解和调控农田微生物群体组成以促进秸秆分解具有重要意义。【方法】试验于2014年10月至2015年10月在河南省农业科学院原阳试验基地进行,将成熟期玉米秸秆(茎和叶)烘干,剪成长1~2 cm、宽0.3~1 cm的碎片,称12 g样品(相当于8 t/hm^2)装入15 cm×10 cm的尼龙网包(孔径0.04 mm)内,于10月5日冬小麦出苗后埋置在小麦垄间。分别于埋置后0、1、2、4、7、10和12个月收集秸秆包和土壤样品。测定秸秆样品干物质量和碳氮含量,选择埋置了0、2、4、7和12个月的秸秆及其土壤样品分析细菌丰度及群落组成。【结果】秸秆埋入土壤后的前2个月内分解最快,然后逐步减慢,在1、2、4、7、10和12个月后分别降解了总生物量的19.2%、32.9%、44.2%、52.2%、66.8%和73.8%。秸秆埋入土壤后,秸秆和土壤中细菌丰度均显著增加,分别于第4和7个月达到最高后开始下降。秸秆细菌的丰度指标OTUs、ACE、Chao1和多样性指标Shannon随试验时间的延长逐步增加,而Simpson指数随试验时间延长逐步降低,而土壤中这些指标在试验过程中没有显著变化。与刚埋置秸秆时相比,埋置2个月后的秸秆细菌Bacteroidetes门相对丰度明显增加,主导细菌群为Bacteroidetes和Proteobacteria门。Actinobacteria丰度在埋置2个月后明显降低,然后又随试验时间延长逐步增加。Planctomycetes、Saccharibacteria、Verrucomicrobia、Acidobacteria、Chloroflexi和Gemmatimonadetes丰度在原始秸秆中较低,埋入土壤后随试验时间延长逐步增加。Sphingobacteriia、Gammaproteobacteria、Alphaproteobacteria和Flavobacteriia主导前期细菌纲组成,而Actinobacteria、Anaerolineae和Bacilli纲丰度在后期逐步增加。秸秆分解速率主要受其碳含量影响,秸秆细菌群落组成前期与秸秆碳含量相关,后期与秸秆氮含量相关。随着试验的进展,秸秆细菌群落组成与土壤中的细菌群落组成趋同。【结论】秸秆埋入土壤后前2个月的分解速率最高,随后逐步降低。秸秆分解前期细菌群落由富营养型组分Bacteroidetes和Proteobacteria门和Sphingobacteriia、Gammaproteobacteria、Flavobacteriia和Alphaproteobacteria纲主导,随后被逐步增加的贫营养型组分Actinobacteria、Acidobacteria、Chloroflexi、Saccharibacteria门和Deltaproteobacteria、Actinobacteria纲等代替。秸秆碳氮含量变化是影响秸秆分解及其过程中细菌群落演化的主要原因。     

原料奶中微生物区系ERIC-PCR DNA指纹图谱的建立

为了研究原料奶中微生物区系的结构,该研究采用肠杆菌基因间保守重复序列ERIC-PCR技术,建立了华北和中南地区原料奶中微生物区系的ERIC-PCR DNA指纹图谱。为获得清晰且高质量的图谱,对ERIC-PCR的反应条件进行了优化,确定最佳条件为：退火温度52℃、MgCl2 1 μL、引物0.6 μL、Taq酶0.5 μL。对2个地区指纹图谱进行综合分析鉴定。结果显示,地理位置距离较近即相近地域原料奶样品其微生物群落的构成相似性较高,存在一定特征性优势菌群;距离较远则相似性较低。研究结果提示,建立中国不同地区原料奶微生物区系DNA的特征图谱,将为原料奶质量与安全的监测提供参考。     

Leaf litter is the main driver for changes in bacterial community structures in the rhizosphere of ash and beech

The rhizosphere and the surrounding soil harbor an enormous microbial diversity and a specific community structure, generated by the interaction between plant roots and soil bacteria. The aim of this study was to address the influences of tree species, tree species diversity and leaf litter on soil bacterial diversity and community composition. Therefore, mesocosm experiments using beech, ash, lime, maple and hornbeam were established in 2006, and sampled in October 2008 and June 2009. Mesocosms were planted with one, three or five different tree species and treated with or without litter overlay.Cluster analysis of DGGE-derived patterns revealed a clustering of 2008 sampled litter treatments in two separated clusters. The corresponding treatments sampled in 2009 showed separation in one cluster. PCA analysis based on the relative abundance of active proteobacterial classes and other phyla in beech and ash single-tree species mesocosm indicated an effect of sampling time and leaf litter on active bacterial community composition. The abundance of next-generation sequencing-derived sequences assigned to the Betaproteobacteria was higher in the litter treatments, indicating a higher activity, under these conditions. The Deltaproteobacteria, Nitrospira and Gemmatimonadetes showed an opposite trend and were more active in the mesocosms without litter. The abundance of alphaproteobacterial sequences was higher in mesocosms sampled in 2009 (P = 0.014), whereas the Acidobacteria were more active in 2008 (P = 0.014). At the family level, we found significant differences of the litter vs. non-litter treated group. Additionally, an impact of beech and ash as tree species on soil bacterial diversity was confirmed by the Shannon and Simpson indices. Our results suggest that leaf litter decomposition in pH-stable soils affect the soil bacterial composition, while tree species influence the soil bacterial diversity.     

基于高通量测序的牡蛎冷藏过程中微生物群落分析

为揭示牡蛎冷藏过程中微生物群落结构的变化规律,采用高通量测序技术对冷藏0、4、8 d牡蛎的菌群进行分析。结果表明高通量测序可以全面反映牡蛎中菌群的物种组成及丰度信息。牡蛎体附着的细菌以变形菌门(Proteobacteria)、γ-变形菌纲(Gammaproteobacteria)、弧菌目(Vibrionales)为主。在科的分类水平上,主要是弧菌科(Vibrionaceae)、希瓦氏菌科(Shewanellaceae)和交替假单胞菌科(Pseudoalteromonadaceae),其中初始样品以Vibrionaceae为主,比例达36.8%;第4天样品Pseudoalteromonadaceae比例相对较高;第8天样品同样以Pseudoalteromonadaceae为主,而Shewanellaceae比例明显增加,达到19.5%。在属的分类水平上,在牡蛎初始菌群中比例前三位依次为弧菌属(Vibrio)、希瓦氏菌属(Shewanella)和交替假单胞菌属(Pseudoalteromonas),其中Vibrio在冷藏前期比例迅速下降;Shewanella和Pseudoalteromonas在系统进化关系上较为接近,在冷藏后期比例占优,可能在牡蛎腐败过程中起到重要作用。研究结果可以为牡蛎的贮运保鲜策略提供参考。     

贮藏温度对冷破碎番茄浆品质变化的影响

为探讨冷破碎番茄浆在贮藏过程中的品质变化,以指导冷破碎番茄浆的贮藏,该文研究了其在0、25和37℃条件下贮藏150 d的品质变化,并对产品中主要指标的变化进行了动力学分析。25和37℃贮藏条件下冷破碎番茄浆中氨基酸、总糖、蔗糖、果糖、维生素C含量和颜色指标（亮度L*、红值a*和黄值b*）均显著下降,而5-羟甲基糠醛（HMF）含量、褐变度（BD）和总色差ΔE显著增加,其中总氨基酸、总糖、维生素C、HMF和BD的变化均符合一级动力学模型;而且HMF含量与BD变化呈线性关系,符合零级动力学模型。但是番茄红素和可溶性固形物含量变化不显著,而酸度随温度和时间的增加呈增加趋势。0℃贮藏条件下,除维生素C含量显著下降外,上述其他指标均无显著变化。表明高温贮藏显著降低冷破碎番茄浆的品质,而低温贮藏可有效保护冷破碎番茄浆的品质。     

芦苇定居后铜尾矿细菌群落结构的变化

Soil samples were collected from both bare and vegetated mine tailings to study the changes in bacterial communities and soil chemical properties of copper mine tailings due to reed (Phragmites communis) colonization. The structures of bacterial communities were investigated using culture-independent 16S rRNA gene sequencing method. The bacterial diversity in the bare mine tailing was lower than that of the vegetated mine tailing. The former was dominated by sulfur metabolizing bacteria, whereas the latter was by nitrogen fixing bacteria. The bare mine tailing was acidic (pH = 3.78), whereas the vegetated mine tailing was near neutral (pH = 7.28). The contents of organic matter, total nitrogen, and ammonium acetate-extractable potassium in vegetated mine tailings were significantly higher than those in the bare mine tailings (P < 0.01), whereas available phosphorus and electrical conductivity were significantly lower than those in the bare mine tailings (P < 0.01). The results demonstrated that 16S rRNA gene sequencing could be successfully used to study the bacterial diversity in mine tailings. The colonization of the mine tailings by reed significantly changed the bacterial community and the chemical properties of tailings. The complex interactions between bacteria and plants deserve further investigation.     

Rapid changes in the rhizosphere bacterial community structure during re-colonization of sterilized soil

Diversity has been shown to be pivotal in ecosystem stability and resilience. It is therefore important to increase our knowledge about the development of diversity. The aim of this study was to investigate the temporal dynamics of the bacterial community structure in the rhizosphere of wheat plants growing in a soil in which the initial conditions for bacterial re-colonization were modified by mixing different amounts of sterilized with native soil at ratios of 19:1, 9:1, 4:1 and 1:1. Additional treatments comprised sterilized soil or native soil. Plant dry weight at day 20 decreased with increasing percentage of native soil in the mix. The bacterial community structure in the rhizosphere was assessed by polymerase chain reaction-denaturing gradient gel electrophoresis (DGGE) at days 3, 14 and 20 after planting. The bacterial community in the sterilized soil had a lower diversity and evenness than the native soil. Both diversity and evenness increased with time in the sterilized soil. Community structure in the different mixes changed over time and the changes were mix-specific. Principal component analyses of the DGGE banding patterns showed clear differences between the treatments particularly at day 3 and day 14 and revealed changes in community structure within a few days in a given treatment. The results of the present study show that bacterial communities rapidly re-colonize sterilized soil. During re-colonization, the community structure changes rapidly with a general trend towards higher diversity and evenness. The changes in community structure over time are also affected by the amount of sterile substrate to be re-colonized.