Advances in high-throughput sequencing (HTS) have fostered rapid developments in the field of microbiome research, and massive microbiome datasets are now being generated. However, the diversity of software tools and the complexity of analysis pipelines make it difficult to access this field. Here, we systematically summarize the advantages and limitations of microbiome methods. Then, we recommend specific pipelines for amplicon and metagenomic analyses, and describe commonly-used software and databases, to help researchers select the appropriate tools. Furthermore, we introduce statistical and visualization methods suitable for microbiome analysis, including alpha- and beta-diversity, taxonomic composition, difference comparisons, correlation, networks, machine learning, evolution, source tracing, and common visualization styles to help researchers make informed choices. Finally, a step-by-step reproducible analysis guide is introduced. We hope this review will allow researchers to carry out data analysis more effectively and to quickly select the appropriate tools in order to efficiently mine the biological significance behind the data.

Mix of metabolites tunes root microbiota: Uncharacterized biosynthetic genes in plant genomes suggest that plants make a plethora of specialized metabolites. Huang et al. reconstructed three biosynthetic networks from the small mustard plant Arabidopsis thaliana . Promiscuous acyltransferases and dehydrogenases contributed to metabolite diversification. The plant may use these specialized metabolites to modulate the microbiota surrounding its roots. Disruption of the pathways and intervention with purified compounds caused changes in the root microbiota. Science , this issue p. eaau6389

The network analysis has attracted increasing attention and interest from ecological academics, thus it is of great necessity to develop more convenient and powerful tools. For that reason, we have developed an R package, named “ggClusterNet,” to complete and display the network analysis in an easier manner. In that package, ten network layout algorithms are designed to better display the modules of microbiome network (randomClusterG, PolygonClusterG, PolygonRrClusterG, ArtifCluster, randSNEClusterG, PolygonModsquareG, PolyRdmNotdCirG, model_Gephi.2, model_igraph, and model_maptree). For the convenience of the users, many functions related to microbial network analysis, such as corMicor(), net_properties(), node_properties(), ZiPiPlot(), random_Net_compate(), are integrated to complete the network mining. Furthermore, the pipeline function named network.2() and corBionetwork() are also added for the quick achievement of the network or bipartite network analysis as well as their in‐depth mining. The ggClusterNet is publicly available via GitHub (https://github.com/taowenmicro/ggClusterNet/) or Gitee (https://gitee.com/wentaomicro/ggClusterNet) for users' access. A complete description of the usages can be found on the manuscript's GitHub page (https://github.com/taowenmicro/ggClusterNet/wiki).