Bacillus Amyloliquefaciens SQR9 is a well-investigated plant growth-promoting rhizobacteria with strong root colonization capability. To identify the key proteins involved in in situ root colonization and biofilm formation, the proteomic profiles of planktonic and root colonized SQR9 cells were compared. A total of 755 proteins were identified, of which 78 and 95 proteins were significantly increased and deceased, respectively, when SQR9 was colonized on the root. The proteins that were closely affiliated with the root colonization belonged to the functional categories of biocontrol, detoxification, biofilm formation, cell motility and chemotaxis, transport, and degradation of plant polysaccharides. A two-component system protein ResE was increased 100-fold when compared to the planktonic status; impairment of the resE gene postponed the formation of cell biofilm and decreased the root colonization capability, which may be regulated through the spo0A-sinI-yqxM pathway. The SQR9 proteomic data provide valuable clues for screening key proteins in the plant–rhizobacteria interaction.Keywords: biofilm formation; proteomics; resE; root colonization;

•Bacterial communities operate better in higher-productivity soils.•Networks of higher-productivity soils contain more module hubs.•Soil environmental conditions altered the topological roles of OTUs.•Module hubs could be indicators of soil productivity.Interactions among soil bacteria occur widely and play important roles in the maintenance of soil functions. Long-term fertilization management practices have distinct effects on soil fertility and the soil microbial activity and community, which are closely associated with soil microbial interactions. Red soil is typically low-productivity soil in South China. Chemical nitrogen fertilization caused serious soil acidification and low-productivity (defined as the acidified soil group, Ac), whereas the application of lime to the acidified soil increased the soil pH (defined as the quicklime improvement soil group, Qlime). Long-term manure or fallow treatment maintained the soil pH and increased the soil fertility (defined as the high-productivity potential soil group, HPP). A molecular ecological network analysis method was used to analyze 454 pyrosequencing data of bacterial communities from the HPP, Ac and Qlime soils. Several major differences were observed among the three constructed networks. First, the HPP network contained the largest ratio of positive to negative correlations, whereas the Ac network contained the smallest. Second, the HPP and Ac networks shared only 8.67% of their operational taxonomic units (OTUs), whereas Ac and Qlime shared 27.04%. Third, the HPP network contained the most “module hubs” (A set of OTUs that have strong interactions or common functions are grouped as a “module” in network analysis. These OTUs are called “nodes”. And the nodes with high connectivity to many other nodes within the same module are “module hubs”.), whereas Ac contained the fewest. These results demonstrated that the bacterial community of HPP was a better organized or a better operated community than Ac and that quicklime application helped to order the bacterial community. By comparing the topological roles of nodes in different networks, we proposed that there should be more module hubs in the networks of higher-productivity soils and hypothesized that these OTUs could be indicators of high-productivity.

Natural ecosystems comprise the planet’s wild plant and animal resources, but large tracts of land have been converted to agroecosystems to support the demand for agricultural products. This conversion limits the number of plant species and decreases the soil biological diversity. Here we used high-throughput 16S rRNA gene sequencing to evaluate the responses of soil bacterial communities in long-term converted and fertilized red soils (a type of Ferralic Cambisol). We observed that soil bacterial diversity was strongly affected by different types of fertilization management. Oligotrophic bacterial taxa demonstrated large relative abundances in chemically fertilized soil, whereas copiotrophic bacterial taxa were found in large relative abundances in organically fertilized and fallow management soils. Only organic-inorganic fertilization exhibited the same local taxonomic and phylogenetic diversity as that of a natural ecosystem. However, the independent use of organic or inorganic fertilizer reduced local taxonomic and phylogenetic diversity and caused biotic homogenization. This study demonstrated that the homogenization of bacterial communities caused by natural-to-agricultural ecosystem conversion can be mitigated by employing rational organic-inorganic fertilization management.

Bacillus amyloliquefaciens SQR9, isolated from the rhizosphere of cucumber, can control Fusarium wilt of cucumber and directly stimulate plant growth. To evaluate its potential agricultural use, the plant growth promotion of B. amyloliquefaciens SQR9 was evaluated, and the relative mechanisms, especially the production of the phytohormone indole-3-acetic acid (IAA), were investigated. The related plant-growth-promoting factors were genetically and chemically analyzed, and a mutant library was constructed for selecting strains with different IAA production. B. amyloliquefaciens SQR9 showed a growth-promoting activity in greenhouse experiments. Plant-growth-promoting factors like extracellular phytase, volatile components including acetoin, 2,3-butanediol, and phytohormone IAA were detected in B. amyloliquefaciens SQR9 cultures grown under laboratory conditions. Three IAA production mutant strains showed variation in plant-growth-promoting effect. IAA production in B. amyloliquefaciens SQR9 was related to its plant-growth-promoting effect, but IAA alone could not account for the overall observed plant-growth-promoting effect. The promoted plant growth by the rhizospheric strain B. amyloliquefaciens SQR9 can be attributed to multiple factors, including production of phytohormones, volatile compounds, and extracellular enzymes. Therefore, the strain B. amyloliquefaciens SQR9 may be used as a plant-growth-promoting agent to increase crop yield.

It was hypothesized that disruption of the root–microbiome association creates empty rhizosphere niches that could be filled by both soilborne pathogens and beneficial microbes. The effect of de-coupling root–microbiome associations related to improve soil suppressiveness was investigated in cucumber using the pathogen Fusarium oxysporum f. sp. Cucumerinum (FOC) and its antagonist Bacillus amyloliquefaciens SQR9 (SQR9) system. The root–soil microbiome association of cucumber was disrupted by applying the fungicide carbendazim to the soil, and then FOC or/and its antagonist SQR9 were inoculated in the rhizosphere. In the fungicide treatment, the FOC wilt disease incidence was significantly increased by 13.3 % on average compared to the FOC treatment without fungicide. However, when the fungicide treatment was applied to the soil with SQR9 and FOC, the SQR9 effectively reduced the disease incidence, and improved cucumber plant growth compared to a no fungicide control. These results indicate that de-coupling of root–microbiome associations followed by antagonist inoculation can improve rhizosphere soil suppressiveness, which may help to develop strategies for efficient application of rhizosphere beneficial microbes in agriculture.

Paenibacillus polymyxa SQR-21, which is antagonistic against Fusarium oxysporum, is used as a biocontrol agent and, when mixed with organic substances for solid fermentation, produces a bioorganic fertilizer. The spores of P. polymyxa prepared at different temperatures were characterized with respect to the dipicolinic acid content, heat resistance, fatty acid composition and germination. Spores prepared at 37°C showed higher heat resistance than those prepared at 25 and 30°C. However, the germination rate was negatively correlated with the sporulation temperature. The maximum germination rate of the spores prepared at 25°C was 1.3-times higher than the spores prepared at 30°C. The sporulation temperature thus affects the resistance and germination properties of P. polymyxa spores. These results are useful for the production of improved bio-organic fertilizer.

•Long-term different fertilization changed soil viral abundance.•Organic fertilization increased soil organic matter and pH value.•Organic fertilization increased soil viral abundance and morphological diversity.Fertilization plays a pivotal role on soil biological process and affects the soil bacterial community, which act as hosts for viruses. The effect of fertilization on soil viral community has not been well explored. In this study, a Haplic Acrisol soil, which is the soil type for 13 provinces in Southern China, was analyzed after 22 years different fertilization regimes for their viral composition. The soil responded to organic fertilizations with an increased amount of soil organic matter (SOM) and pH (increased from 5.7 to 6.6), while with the decreased SOM and pH for chemical fertilization, especially for single nitrogen fertilization. The combined effects of SOM and pH caused by long-term different fertilization regimes on soil viral communities were investigated by direct calculation of virus-like particles (VLPs) through epifluorescence microscopy. The highest VLP abundance (13.1 × 107 per gram dry soil) was detected in soil applied with chemical and organic fertilizers. The viral and bacterial abundances of organic soil were 4 and 5 times higher than those of inorganic soil respectively. Transmission electron microscopy observation revealed a higher frequency of Myoviridae viruses in soil with organic amendments than without organic amendments, and vice versa for Podoviridae viruses. These results demonstrate that organic fertilizer could increase viral abundance and morphological diversity through suppressing soil acidification and improving soil organic matter.

A lignocellulosic decomposing fungus Z5 was isolated and identified as Aspergillus fumigatus, its capacity to produce cellulase was assessed under solid-state fermentation (SSF) using lignocellulosic materials as substrates. Cultivation conditions of A. fumigatus Z5 for cellulase production were optimized, results showed that for carboxymethyl cellulase (CMCase) and filter paper enzyme (FPase), the best condition was 50 °C, 80% initial moisture, initial pH 4.0 and 7% initial inoculum, the average activity of CMCase activity, FPase activity reached 526.3 and 144.6 U g−1 dry weight (dw) respectively, much higher than most of previous reports of this genus. Optimal temperature and pH for the CMCase activity of the crude enzyme were found to be 50 °C and 5.0, respectively. Zymogram analysis showed that eight kinds of CMCase were secreted by A. fumigatus Z5 when cellulose-containing materials were supplied in the culture. The crude enzyme secreted by the strain was further applied to hydrolyze pretreated corn stover and the enzymatic hydrolysate was used as substrate for ethanol production by Saccharomyces cerevisiae. The yield of bio-ethanol was 0.112 g g−1 dry substrate (gDS), suggesting that it is a promising fungus in the bio-ethanol production process.Highlights► A thermostable lignocellulosic decomposing fungus Z5 was isolated and identified. ► Z5 showed high cellulase activity which ranks top of the same genus. ► Zymogram result showed that eight kinds of CMCase were secreted by A. fumigatus Z5. ► The crude enzyme secreted by Z5 was applied to hydrolyze pretreated corn stover. ► The yield of bio-ethanol was 0.112 g g−1 dry substrate (gDS).