宏基因组DNA分析技术在传统发酵乳制品微生物多样性研究中的应用

本文对近几年广泛应用于传统发酵乳制品微生物多样性分析的宏基因组DNA分析技术进行了综述,尤其是对基于16S rRNA基因序列的分子标记技术,如:变性梯度凝胶电泳、限制性片段长度多态性分析、末端标记限制片段长度多态性分析、单链构象多态性等以及这些方法的研究进展.     

水体宏基因组学研究进展及应用

微生物是水体生态系统中不可或缺的一环,占生物基因资源的绝大多数。目前传统的微生物研究手段只能培养0.1%-1%的环境微生物,无法整体分析水体微生物群落的构成和功能,形成了水体治理研究的瓶颈。近年来,不依赖于培养的微生物研究手段即宏基因组学的出现为解决这一难题提供了新方法。一方面宏基因组学技术可以用于研究水体的生物多样性、微生物系统中的物质代谢历程这些为研究水体污染的治理提供了可能;另一方面,用宏基因组学方法从水体中发现新型微生物来源酶具有极高的应用价值。虽然宏基因组学仍存在一些问题和不足之处,但是近年来也取得了巨大的成就,该文综述了宏基因组学的概念、方法、问题和水体宏基因组学研究进展。     

患根肿病的上海青根内生菌组成和结构的研究

旨在优化植物根内生菌高通量测序分析技术,更好地解析健康与患根肿病上海青根内生菌之间的差异。采用高通量测序方法,对其16S rDNA的V3-V4区进行扩增,并通过合适的软件和数据库进行菌群分析。结果表明,植物内生菌受植物叶绿体DNA的干扰大,平均叶绿体数据占67%。上海青根内生微生物菌群具有丰富的多样性,主要由变形菌门、放线菌门、拟杆菌门和厚壁菌门细菌组成,优势种群为假单胞菌,根瘤菌,芽孢杆菌,以及链霉菌等。健康样品的稳定性较好,不同患根肿病样品之间菌群比例差异较大。研究显示,根肿病侵染对上海青根内生菌群的结构和组成有较为显著的影响。     

宏基因组法研究不同堆肥处理下猪粪微生物动态结构特征

试验旨在应用宏基因组测序技术探究两种堆肥处理方式下猪粪中微生物的结构组成与分布特征。试验采集同一粪池中的新鲜猪粪，随机分成添加有效微生物（EM）菌的F组和未添加EM菌的K组，每组粪堆各1堆，堆成圆锥形粪堆，粪堆高1.2 m，底部直径2 m；采集堆肥第0、7、14和21天的粪便样本并提取粪便总DNA，运用宏基因组高通量测序技术，比较F组和K组的微生物多样性（组成及结构）差异，并对功能基因进行注释。结果表明，猪粪便中的细菌共测序获得1 083 607个有效开放阅读框（ORFs）；共鉴定出147个门、118个纲、258个目、593个科、2 343个属和13 193个种；在门水平上，相对丰度前十的细菌界优势菌门有：变形菌门（Proteobacteria）、拟杆菌门（Bacteroidetes）、放线菌门（Actinobacteria）、厚壁菌门（Firmicutes）、疣微菌门（Verrucomicrobia）、绿弯菌门（Chloroflexi）、柔壁菌门（Tenericutes）、浮霉菌门（Planctomycetes）、螺旋体门（Spirochaetes）和酸杆菌门（Acidobacteria）；在属水平上，两组平均丰度值排名前三十五的属分别属于变形菌门、拟杆菌门、厚壁菌门、放线菌门和疣微菌门；通过KEGG功能预测分析，新陈代谢通路是最丰富的，氨基酸代谢通路和碳水化合物代谢通路是包含功能基因数量最多的二级通路。两组的菌群结构多样性差异表明添加EM菌后会促进堆肥前期的菌群变化。     

Identification of rumen microbial biomarkers linked to methane emission in Holstein dairy cows

Mitigation of greenhouse gas emissions is relevant for reducing the environmental impact of ruminant production. In this study, the rumen microbiome from Holstein cows was characterized through a combination of 16S rRNA gene and shotgun metagenomic sequencing. Methane production (CH₄) and dry matter intake (DMI) were individually measured over 4–6 weeks to calculate the CH₄ yield (CH₄y = CH₄/DMI) per cow. We implemented a combination of clustering, multivariate and mixed model analyses to identify a set of operational taxonomic unit (OTU) jointly associated with CH₄y and the structure of ruminal microbial communities. Three ruminotype clusters (R1, R2 and R3) were identified, and R2 was associated with higher CH₄y. The taxonomic composition on R2 had lower abundance of Succinivibrionaceae and Methanosphaera, and higher abundance of Ruminococcaceae, Christensenellaceae and Lachnospiraceae. Metagenomic data confirmed the lower abundance of Succinivibrionaceae and Methanosphaera in R2 and identified genera (Fibrobacter and unclassified Bacteroidales) not highlighted by metataxonomic analysis. In addition, the functional metagenomic analysis revealed that samples classified in cluster R2 were overrepresented by genes coding for KEGG modules associated with methanogenesis, including a significant relative abundance of the methyl-coenzyme M reductase enzyme. Based on the cluster assignment, we applied a sparse partial least-squares discriminant analysis at the taxonomic and functional levels. In addition, we implemented a sPLS regression model using the phenotypic variation of CH₄y. By combining these two approaches, we identified 86 discriminant bacterial OTUs, notably including families linked to CH₄ emission such as Succinivibrionaceae, Ruminococcaceae, Christensenellaceae, Lachnospiraceae and Rikenellaceae. These selected OTUs explained 24% of the CH₄y phenotypic variance, whereas the host genome contribution was ~14%. In summary, we identified rumen microbial biomarkers associated with the methane production of dairy cows; these biomarkers could be used for targeted methane-reduction selection programmes in the dairy cattle industry provided they are heritable.     

Integrated analysis of changes in soil microbiota and metabolites following long-term fertilization in a subtropical maize-wheat agroecosystem

Although the application of inorganic fertilizer is a widespread agronomic practice used to boost soil productivity and crop yields, the effects on soil microbiome and the metabolic mechanisms involved in the high-yield response of crops to long-term fertilization remain poorly described. In this study, combined metagenomic and metabolomic analyses were used to explore the mechanism of crop yield response to the 20-year application of nitrogen-phosphorus-potassium fertilizers in a subtropical ag...     

Antiviral lead compounds from marine sponges

Marine sponges are currently one of the richest sources of pharmacologically active compounds found in the marine environment. These bioactive molecules are often secondary metabolites, whose main function is to enable and/or modulate cellular communication and defense. They are usually produced by functional enzyme clusters in sponges and/or their associated symbiotic microorganisms. Natural product lead compounds from sponges have often been found to be promising pharmaceutical agents. Several of them have successfully been approved as antiviral agents for clinical use or have been advanced to the late stages of clinical trials. Most of these drugs are used for the treatment of human immunodeficiency virus (HIV) and herpes simplex virus (HSV). The most important antiviral lead of marine origin reported thus far is nucleoside Ara-A (vidarabine) isolated from sponge Tethya crypta. It inhibits viral DNA polymerase and DNA synthesis of herpes, vaccinica and varicella zoster viruses. However due to the discovery of new types of viruses and emergence of drug resistant strains, it is necessary to develop new antiviral lead compounds continuously. Several sponge derived antiviral lead compounds which are hoped to be developed as future drugs are discussed in this review. Supply problems are usually the major bottleneck to the development of these compounds as drugs during clinical trials. However advances in the field of metagenomics and high throughput microbial cultivation has raised the possibility that these techniques could lead to the cost-effective large scale production of such compounds. Perspectives on biotechnological methods with respect to marine drug development are also discussed.     

瘤胃厌氧真菌的作用及其分子生物学研究方法

瘤胃厌氧真菌这一特殊的菌群除在瘤胃中发挥重要作用外,其分泌的高活性纤维素酶的应用价值也备受关注,因传统培养的局限性,因此,利用分子生物技术开展的研究逐渐深入。作者主要从瘤胃厌氧真菌的特点和多样性出发,厌氧真菌对结构性碳水化合物、淀粉和蛋白质的降解及在瘤胃生态环境中的作用进行了分析,并对研究瘤胃厌氧真菌的分子生物学方法如内转录间隔区(ITS)、变性梯度凝胶电泳(DGGE)、限制性片段长度多态性(RFLP)、实时荧光定量PCR(Real-time PCR,RT-PCR)和宏基因组学(metagenomics)等进行了阐述,以期为瘤胃厌氧真菌的研究提供参考。     

宏基因组学在动物胃肠道微生物研究中的应用

实验室条件下可培养的微生物约占自然界中微生物总数的1%,这限制了人们对环境微生物的了解和利用.宏基因组学绕开传统纯培养方法,提供了一种探知微生物多样性和功能基因组的新方法.作者简述了宏基因组学的概念、基本方法,着重介绍了宏基因组学在动物胃肠道微生物研究中的应用和取得的成果,并对宏基因组学的发展做了简要分析.