Combined application of microbial inoculant and kelp-soaking wastewater promotes wheat seedlings growth and improves structural diversity of rhizosphere microbial community.

Industrial processing of kelp generates large amounts of kelp-soaking wastewater (KSW), which contains a large amount of nutrient-containing substances. The plant growth-promoting effect might be further improved by combined application of growth-promoting bacteria and the nutrient-containing KSW. Here, a greenhouse experiment was conducted to determine the effect of the mixture of KSW and Bacillus methylotrophicus M4-1 (MS) vs. KSW alone (SE) on wheat seedlings, soil properties and the microbial community structure in wheat rhizosphere soil. The available potassium, available nitrogen, organic matter content and urease activity of MS soil as well as the available potassium of the SE soil were significantly different (p < 0.05) from those of the CK with water only added, increased by 39.51%, 36.25%, 41.61%, 80.56% and 32.99%, respectively. The dry and fresh weight of wheat seedlings from MS plants increased by 166.17% and 50.62%, respectively, while plant height increased by 16.99%, compared with CK. Moreover, the abundance and diversity of fungi in the wheat rhizosphere soil were significantly increased (p < 0.05), the relative abundance of Ascomycetes and Fusarium spp. decreased, while the relative abundance of Bacillus and Mortierella increased. Collectively, the combination of KSW and the plant growth-promoting strain M4-1 can promote wheat seedlings growth and improve the microecology of rhizosphere microorganisms, thereby solving the problems of resource waste and environmental pollution, ultimately turning waste into economic gain.

Bacillus subtilis HG-15, a Halotolerant Rhizoplane Bacterium, Promotes Growth and Salinity Tolerance in Wheat (Triticum aestivum).

Certain plant growth-promoting bacteria (PGPB) reduce salt stress damage in plants. Bacillus subtilis HG-15 is a halotolerant bacterium (able to withstand NaCl concentrations as high as 30%) isolated from the wheat rhizoplane in the Yellow River delta. A qualitative and quantitative investigation of the plant growth-promoting characteristics of this strain confirmed nitrogen fixation, potassium dissolution, ammonia, plant hormone, ACC deaminase, and proline production abilities. B. subtilis HG-15 colonization of wheat roots, stems, and leaves was examined via scanning electron microscopy, rep-PCR, and double antibiotic screening. After inoculation with the B. subtilis HG-15 strain, the pH (1.08-2.69%), electrical conductivity (3.17-11.48%), and Na+ (12.98-15.55%) concentrations of rhizosphere soil significantly decreased (p < 0.05). Under no-salt stress (0.15% NaCl), low-salt stress (0.25% NaCl), and high-salt stress (0.35% NaCl) conditions, this strain also significantly increased (p < 0.05) the dry weight (17.76%, 24.46%, and 9.31%), fresh weight (12.80%, 20.48%, and 7.43%), plant height (7.79%, 5.86%, and 13.13%), and root length (10.28%, 17.87%, and 48.95%). Our results indicated that B. subtilis HG-15 can effectively improve the growth of wheat and elicit induced systemic tolerance in these plants, thus showing its potential as a microbial inoculant that can protect wheat under salt stress conditions.

Bacillus licheniformis JF-22 to Control Meloidogyne incognita and Its Effect on Tomato Rhizosphere Microbial Community.

Meloidogyne incognita is one of the most destructive soil pests, causing serious economic losses in tomato production. Here, in vitro experiments demonstrated that the Bacillus licheniformis strain JF-22 has the potential to prevent M. incognita infection. A pot experiment confirmed that B. licheniformis strain JF-22 isolated from the tomato rhizosphere soil and planted in the tomato root-knot nematode disease area effectively prevented and controlled M. incognita, reducing its negative effect on tomato growth. Additionally, the composition of volatile substances secreted by B. licheniformis strain JF-22 was analyzed using solid-phase microextraction and gas chromatography-mass spectrometry. We detected acetoin, 2,3-Butanediol, [R-(R*,R*) ]-, and hexamethyl cyclotrisiloxane as the main components among these volatiles. Using MiSeq sequencing technology and bioinformatics, we analyzed the influence of B. licheniformis strain JF-22 on the microbial community of the tomato rhizosphere. B. licheniformis strain JF-22 changed the composition of the microbial community; particularly, it significantly reduced the diversity of the fungal community. Furthermore, using the FUNGuild and PICRUSt databases, we predicted the effect of JF-22 on microbial community function. In conclusion, B. licheniformis strain JF-22 may be considered as a potential biocontrol agent against M. incognita.

Impacts of continuous and rotational cropping practices on soil chemical properties and microbial communities during peanut cultivation.

Long-term monocultures have severely inhibited the cultivation of Chinese peanut (Arachis hypogaea L.). In this study, the effects of continuous cropping on soil chemical properties and microbial communities were investigated in peanut fields that had been in crop rotation for 10 years and in monoculture for 10 years. The results found that long-term monoculture increased the activities of available potassium, available phosphorus, available nitrogen, soil organic matter, urease, acid phosphatase and catalase; while decreasing the activity of catalase. The diversity and abundance of soil bacteria and fungi is higher under continuous peanut cultivation. At the genus level, the relative abundance of potentially beneficial microflora genera was higher in the rhizosphere soil of rotational cropping than in continuous cropping, while the opposite was true for the relative abundance of potentially pathogenic fungal genera. Principal coordinates and cluster analysis indicated that continuous cropping altered the structure of the microbial community. The results of the functional predictions showed significant differences in the functioning of the rhizosphere microbial community between continuous and rotational cropping. In conclusion, long-term continuous cropping changed the chemical properties of the soil, altered the structure and function of the soil bacterial and fungal communities in peanut rhizosphere, which to some extent reduced the relative abundance of potentially beneficial microbial genera and increased the relative abundance of potentially pathogenic fungal genera, thus increasing the potential risk of soil-borne diseases and reducing the yield and quality of peanut. Therefore, in the actual production process, attention should be paid not only to the application of chemical fertilizers, but also to crop rotation and the application of microbial fertilizers.

The Effect of Salt-Tolerant Antagonistic Bacteria CZ-6 on the Rhizosphere Microbial Community of Winter Jujube (Ziziphus jujuba Mill. "Dongzao") in Saline-Alkali Land.

As the main economic crop cultivated in the Yellow River Delta, winter jujube contains various nutrients. However, soil salinization and fungal diseases have affected the yield and quality of winter jujube. In order to use plant growth-promoting rhizobacteria (PGPR) to reduce these damages, the antagonistic bacteria CZ-6 isolated from the rhizosphere of wheat in saline soil was selected for experiment. Gene sequencing analysis identified CZ-6 as Bacillus amyloliquefaciens. In order to understand the salt tolerant and disease-resistant effects of CZ-6 strain, determination of related indicators of salt tolerance, pathogen antagonistic tests, and anti-fungal mechanism analyses was carried out. A pot experiment was conducted to evaluate the effect of CZ-6 inoculation on the rhizosphere microbial community of winter jujube. The salt tolerance test showed that CZ-6 strain can survive in a medium with a NaCl concentration of 10% and produces indole acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase. Studies on the inhibition mechanism of pathogenic fungi show that CZ-6 can secrete cellulase, protease, and xylanase. Gas chromatography-mass spectrometry (GC-MS) analysis showed that CZ-6 can release volatile organic compounds (VOCs), including 2-heptanone and 2-nonanone. In addition, the strain can colonize the rhizosphere and migrate to the roots, stems, and leaves of winter jujube, which is essential for plant growth or defense against pathogens. Illumina MiSeq sequencing data indicated that, compared to the control, the abundance of salt-tolerant bacteria Tausonia in the CZ-6 strain treatment group was significantly increased, while the richness of Chaetomium and Gibberella pathogens was significantly reduced. Our research shows that CZ-6 has the potential as a biological control agent in saline soil. Plant damage and economic losses caused by pathogenic fungi and salt stress are expected to be alleviated by the addition of salt-tolerant antagonistic bacteria.

Biocontrol of Two Bacterial Inoculant Strains and Their Effects on the Rhizosphere Microbial Community of Field-Grown Wheat.

Biocontrol by inoculation with beneficial microbes is a proven strategy for reducing the negative effect of soil-borne pathogens. We evaluated the effects of microbial inoculants BIO-1 and BIO-2 in reducing soil-borne wheat diseases and in influencing wheat rhizosphere microbial community composition in a plot test. The experimental design consisted of three treatments: (1) Fusarium graminearum F0609 (CK), (2) F. graminearum + BIO-1 (T1), and (3) F. graminearum F0609 + BIO-2 (T2). The results of the wheat disease investigation showed that the relative efficacies of BIO-1 and BIO-2 were up to 82.5% and 83.9%, respectively. Illumina MiSeq sequencing revealed that bacterial abundance and diversity were significantly higher (P < 0.05) in the treatment groups (T1 and T2) than in the control, with significantly decreased fungal diversity in the T2 group. Principal coordinates and hierarchical clustering analyses revealed that the bacterial and fungal communities were distinctly separated between the treatment and control groups. Bacterial community composition analysis demonstrated that beneficial microbes, such as Sphingomonas, Bacillus, Nocardioides, Rhizobium, Streptomyces, Pseudomonas, and Microbacterium, were more abundant in the treatment groups than in the control group. Fungal community composition analysis revealed that the relative abundance of the phytopathogenic fungi Fusarium and Gibberella decreased and that the well-known beneficial fungi Chaetomium, Penicillium, and Humicola were more abundant in the treatment groups than in the control group. Overall, these results confirm that beneficial microbes accumulate more easily in the wheat rhizosphere following application of BIO-1 and BIO-2 and that the relative abundance of phytopathogenic fungi decreased compared with that in the control group.

Effect ofBacillus velezensis JC-K3 on Endophytic Bacterial and Fungal Diversity in Wheat Under Salt Stress.

Plant growth-promoting bacteria (PGPB) can effectively reduce salt damage in plants. Currently, there are many studies on the effects of PGPB on the microbial community structure of rhizosphere soil under salt stress, but fewer studies on the community structure of endophytic bacteria and fungi. We propose that inoculation of endophytic bacteria into the rhizosphere of plants can significantly affect the microbial community structure of the plant's above-ground and underground parts, which may be the cause of the plant's "Induced Systemic Tolerance." The isolated endophytes were re-inoculated into the rhizosphere under salinity stress. We found that, compared with the control group, inoculation with endophytic Bacillus velezensis JC-K3 not only increased the accumulation of wheat biomass, but also increased the content of soluble sugar and chlorophyll in wheat, and reduced the absorption of Na in wheat shoots and leaves. The abundance of bacterial communities in shoots and leaves increased and the abundance of fungal communities decreased after inoculation with JC-K3. The fungal community richness of wheat rhizosphere soil was significantly increased. The diversity of bacterial communities in shoots and leaves increased, and the richness of fungal communities decreased. JC-K3 strain improved wheat's biomass accumulation ability, osmotic adjustment ability, and ion selective absorption ability. In addition, JC-K3 significantly altered the diversity and abundance of endophytic and rhizosphere microorganisms in wheat. PGPB can effectively reduce plant salt damage. At present, there are many studies on the effect of PGPB on the microbial community structure in rhizosphere soil under salt stress, but there are few studies on the community structure changes of endophytic bacteria and fungi in plants.

Effects of Enterobacter cloacae HG-1 on the Nitrogen-Fixing Community Structure of Wheat Rhizosphere Soil and on Salt Tolerance.

The present study investigated the physiological and biochemical characteristics of Enterobacter cloacae HG-1 isolated from saline-alkali soil. We further studied the effect of this strain on the salt tolerance of wheat and on the community structure of nitrogen-fixing bacteria in rhizosphere soil. We determined that the investigated strain had high nitrogen fixation activity and produced iron carriers, 1-aminocyclopropane-1-carboxylic acid deaminase, and plant hormones. The metabolites of this strain contained 2,3-butanediol, [R-(R*, R*)], 2-heptanone, and other growth-promoting and antibacterial substances. The strain was also highly salt-tolerant (10% NaCl). After the inoculation of wheat with the HG-1 strain, we recorded increases in root length, plant height, fresh weight, and dry weight of 19.15%, 18.83%, 16.67%, and 17.96%, respectively, compared with uninoculated plants (P < 0.05). Compared with the leaves of uninoculated plants, the proline concentration in the leaves of inoculated plants increased by 12.43% (P < 0.05), the malondialdehyde level decreased by 27.26% (P < 0.05), K+ increased by 20.69%, Ca2+ increased by 57.53% and Na+ decreased by 31.43% (all P<0.05). Furthermore, we detected that inoculation with the HG-1 strain did not affect the species composition of nitrogen-fixing bacteria in wheat rhizosphere soil at the phylum level. However, the average relative abundance of Proteobacteria was significantly increased, whereas the abundance of Verrucomiorobia was significantly decreased compared with uninoculated plants. At the genus level, we detected 32 genera in control samples and 27 genera in inoculated samples, and the species diversity and relative abundance of samples inoculated with the HG-1 strain decreased compared with uninoculated plants. Inoculated samples had lower abundances of Azospirillum, Rhodomicrobium, and Anabaena. Our study demonstrated that the inoculation of wheat with E. cloacae HG-1 could promote the growth of wheat under salt stress and increase salt stress tolerance. The results of this study investigating the interaction among soil, plants, and microorganisms supplement agricultural microbial databases and could provide a reference for the development of microbial-based saline soil improvement programs.