Optimization of the fermentation media and growth conditions of Bacillus velezensis BHZ-29 using a Plackett-Burman design experiment combined with response surface methodology.

Introduction: Bacillus velezensis occurs extensively in the soil environment. It produces a range of antimicrobial compounds that play an important role in the field of biological control. However, during the actual application process it is often affected by factors such as the medium formulation and fermentation conditions, and therefore biocontrol measures often do not achieve their expected outcomes. Methods: In this study, the B. velezensis BHZ-29 strain was used as the research object. The carbon and nitrogen sources, and inorganic salts that affect the number of viable bacteria and antibacterial potency of B. velezensis BHZ-29, were screened by a single factor test. A Plackett-Burman design experiment was conducted to determine the significant factors affecting the number of viable bacteria and antibacterial potency, and a Box-Behnken design experiment was used to obtain the optimal growth of B. velezensis BHZ-29. The medium formula that produced the highest number of viable bacteria and most antibacterial substances was determined. The initial pH, temperature, amount of inoculant, liquid volume, shaking speed, and culture time were determined by a single factor test. The factors that had a significant influence on the number of viable bacteria of B. velezensis BHZ-29 were selected by an orthogonal test. A Box-Behnken design experiment was conducted to obtain the optimal fermentation conditions, and highest number of viable bacteria and antibacterial titer. Results: Molasses, peptone, and magnesium sulfate had significant effects on the viable count and antibacterial titer of B. velezensis BHZ-29. The viable count of B. velezensis BHZ-29 increased from 7.83 × 109 to 2.17 × 1010 CFU/mL, and the antibacterial titer increased from 111.67 to 153.13 mm/mL when the optimal media were used. The optimal fermentation conditions for B. velezensis BHZ-29 were as follows: temperature 25.57°C, pH 7.23, culture time 95.90 h, rotation speed 160 rpm, amount of inoculant 2%, and liquid volume 100 ml. After the optimization of fermentation conditions, the number of viable bacteria increased to 3.39 × 1010 CFU/mL, and the bacteriostatic titer increased to 158.85 mm/ml.The plant height and leaf number of cotton plants treated with BHZ-29 fermentation broth were higher than those of cotton inoculated with Verticillium dahliae. The number of bacteria was 1.15 × 107 CFU/g, and the number of fungi was 1.60 × 105 spores/g. The disease index of the cotton seedlings treated with the optimized fermentation broth was 2.2, and a control effect of 93.8% was achieved. B. velezensis BHZ-29 could reduce the disease index of cotton Verticillium wilt and had a controlling effect on the disease. The best effect was achieved in the treatment group with an inoculation concentration of 2 × 108 CFU/ml, the disease index was 14.50, and a control effect of 84.18% was achieved. Discussion: The fermentation process parameters of the number of viable bacteria and antibacterial titer by strain B. velezensis BHZ-29 were optimized to lay a foundation for the practical production and application of strain B. velezensis BHZ-29 in agriculture.

Chemical fertilizer reduction combined with organic fertilizer affects the soil microbial community and diversity and yield of cotton.

Introduction: The soil microbial community plays an important role in modulating cotton soil fertility. However, the effects of chemical fertilizer combined with organic fertilizer on soil chemical properties, microbial community structure, and crop yield and quality in arid areas are still unclear. This study aimed to explore the effects of different organic fertilizers on soil microbial community structure and diversity and cotton growth and yield. Methods: High-throughput sequencing was used to study the soil bacteria and fungi in different growth stages of cotton. The field fertilization experiment had five treatments. Results: The results indicated that the treatments of chemical fertilizer reduction combined with organic fertilizer significantly increased soil available nitrogen and phosphorus in cotton field. There were significant differences in the abundance of the bacterial and fungal communities in the dominant phyla among the treatments. At the phyla level, there were not significantly different in the diversity of bacteria and fungi among treatments. There were significant differences in the composition and diversity of bacterial and fungal communities during the entire cotton growth period (p = 0.001). The rhizosphere bacterial and fungal community structure was significantly affected by soil TK, NH4+, AK, TP, AN, and NO3-. The different fertilization treatments strongly influenced the modular structure of the soil bacterial and fungal community co-occurrence network. A reduction in chemical fertilizer combined with organic fertilizer significantly improved cotton stem diameter and seed yield, and the effect of the biological organic fertilizer on plant growth and yield formation was greater than that of ordinary organic fertilizer. Discussion: This study provide a scientific and technical basis for the establishment of environmentally friendly green fertilization technology for cotton in arid areas and the promotion of sustainable development of cotton industry.

Differentiation and Variability in the Rhizosphere and Endosphere Microbiomes of Healthy and Diseased Cotton (Gossypium sp.).

The plant microbiome is a key determinant of health and productivity. However, it is still difficult to understand the structural composition of the bacterial and fungal microbiomes of diseased and healthy plants, especially the spatial dynamics and phylogenies of endophytic and rhizosphere microbial communities. We studied the differentiation and variability in the rhizosphere and endosphere microbiomes of healthy and diseased cotton from north and south of the Tianshan Mountains using the methods of PCR-based high-throughput sequencing and real-time quantitative PCR. The endophytic and rhizosphere bacterial abundances in the diseased plants were greater than those of healthy plants. The numbers of endophytic and rhizosphere fungi associated with diseased plants were greater than those associated healthy plants (p < 0.05). Endophytic and rhizosphere bacteria did not share common OTUs. The dominant rhizosphere bacteria were Proteobacteria (29.70%), Acidobacteria (23.14%), Gemmatimonadetes (15.17%), Actinobacteria (8.31%), Chloroflexi (7.99%), and Bacteroidetes (5.15%). The dominant rhizosphere fungi were Ascomycota (83.52%), Mortierellomycota (7.67%), Basidiomycota (2.13%), Chytridiomycota (0.39%), and Olpidiomycota (0.08%). The distribution of dominant bacteria in different cotton rhizosphere soils and roots differed, with the dominant bacteria Pseudomonas (15.54%) and Pantoea (9.19%), and the dominant fungi Alternaria (16.15%) and Cephalotrichum (9.10%) being present in the greatest numbers. At sampling points in different ecological regions, the total numbers of cotton endophytic and rhizosphere microbiome OTUs from southern to northern Xinjiang showed an increasing trend. There were significant differences in the composition and diversity of rhizosphere microbes and endophytes during the entire cotton growth period and in representative ecological regions (p < 0.01), whereas rhizosphere microbes and endophytes showed no significant differences among the four growth periods and in representative ecological regions. RB41, H16, Nitrospira, and Sphingomonas play important roles in the microbial ecology of cotton rhizosphere soil. Pseudomonas accounted for a large proportion of the microbes in the cotton rhizosphere soil. This study provides an in-depth understanding of the complex microbial composition and diversity associated with cotton north and south of the Tianshan Mountains.

Towards to understanding the preliminary loss and absorption of nitrogen and phosphorus under different treatments in cotton drip- irrigation in northwest Xinjiang.

Drip irrigation under plastic mulch is widely used in Xinjiang, Northwest China. It can not only save water, but also reduce nutrient loss and improve fertilizer utilization. However, it is not clear whether the leaching occurs or not, what is the leaching amount? What is the relationship among fertilization, irrigation regimes, loss, cotton absorption, and cotton field under different fertilization and irrigation management under drip irrigation? Studying these issues not only provides reference for the formulation of fertilization and irrigation systems, but also is of great significance for reducing non-point source pollution. A long-term positioning experiment was conducted from 2009 to 2012 in Baotou Lake farm in Korla City, Xinjiang, with drip-irrigated cotton (Gossypium hirsutum L.) under different N fertilizer and irrigation amounts. The treatments were designed comprising Control (CK,0 N, 0 P, and 0 K with an irrigation of 480 mm) and the following three other treatments: (1) Conventional fertilize and irrigation (CON, 357 kg N hm-2, 90 kg P hm-2, 0 kg K hm-2, and irrigation of 480 mm); (2) Conventional fertilization and Optimizing irrigation (OPT, 357 kg N hm-2, 90 kg P hm-2, 62 kg K hm-2, and irrigation of 420 mm); and (3) Optimizing fertilization and irrigation (OPTN, 240 kg N hm-2, 65 kg P hm-2, 62 kg K hm-2, and irrigation of 420 mm). The results found that the leaching would occur in arid area under drip irrigation. The loss of total N, NH4+, P, N and P loss coefficient was higher under conventional fertilize and irrigation treatment while the loss of NO3- was higher under conventional fertilization and optimizing irrigation treatment. The correlations among N, P absorption by cotton, loss of NH4+ and total phosphorus were quadratic function. The total nitrogen loss and cumulative nitrogen application was lineally correlated. The loss of NO3- and cumulative nitrogen application was exponential. The nitrogen and phosphorus absorption by cotton under conventional fertilization and optimizing irrigation treatment was 24.53% and 35.86% higher than that in conventional fertilize and irrigation treatment, respectively. The cotton yield under conventional fertilization and optimizing irrigation treatment obtained higher than that in other three treatments. Therefore, the conventional fertilization and optimizing irrigation treatment was the optimal management of water and fertilizer in our study. These results demonstrate that reasonable water, nitrogen and phosphorus fertilize could not only effectively promote the absorption of nitrogen and phosphorus, but also reduce nitrogen and phosphorus losses under drip fertigation and plastic mulching.

Diversity and Spatiotemporal Dynamics of Fungal Communities in the Rhizosphere Soil of Cotton in the Arid Region of Northwest China.

This study aimed to investigate the fungal diversity and its temporal and spatial dynamics in the rhizosphere soil of healthy cotton by high-throughput sequencing. We studied species richness, composition, and distribution of cotton rhizosphere fungal community with respect to location (Alaer, Kuerle, Tumushuke, Hami, Shihezi, Wusu, and Jinghe) and plant growth period (seedling stage, bud stage, flowering stage, and boll-opening stage) using the methods of PCR-based high-throughput sequencing and real-time quantitative PCR. A total of 1,838,454 fungal nuclear ribosomal internal transcribed spacer region sequences (rRNA ITS) were obtained from all cotton plants sampled at different growth stages in the seven locations in Xinjiang. The most abundant fungal group in the cotton rhizosphere was the Ascomycota (78.72%), followed by the Zygomycota (9.56%) and Basidiomycota (2.77%). These sequences revealed an enormous number of operational taxonomic units (OTUs) in cotton (1802 unique OTUs), with 67-464 OTUs in a single cotton sample, at a 3% threshold and a sequencing depth of 30,000 sequences. We identified 33 classes and 389 genera from the resulting 1,800,714 sequences. Sordariomycetes was the most frequent class in all samples, followed by Leotiomycetes and Eurotiomycetes. There were some differences in OTUs among different growth stages, but the differences were not significant, with 382 OTUs (14.66%) being common to each of the stages. A marked difference in the diversity of fungi in the rhizosphere soil of cotton was evident among the different locations, with the highest number of OTUs being detected in Jinghe (1084 OTUs) and clusters of OTUs representative of northern and eastern Xinjiang being detected. There were significantly more tags of Mortierella in Jinghe and Wusu than in the other sampling sites. The dynamics of the rhizosphere fungal communities were influenced by sampling sites. To the best of our knowledge, the current study is the first application of PCR-based Illumina to characterize and compare the fungal biodiversity in multiple rhizosphere soil samples from cotton.

Endophytic bacterial communities and spatiotemporal variations in cotton roots in Xinjiang, China.

Endogenous bacteria are important for maintaining the health and other ecologically relevant functions of cotton plants. However, little is known about the community structures and diversity of endophytic bacteria in cotton plants. In our study, we used the Illumina amplicon sequencing technology to study the endophytic bacteria found in cotton root tissue in Xinjiang, China. A total of 60.84 × 106 effective sequences of the 16S rRNA gene in the V5-V6 variable region revealed a large number of operational taxonomic units (OTUs), namely 81-338 OTUs, at a cut-off level of 3% and a sequencing depth of 50 000 sequences. Among the 23 classes identified, Gammaproteobacteria was the dominant group, followed by Alphaproteobacteria, Actinobacteria, and Bacillus. The diversity of endogenous bacteria differed at different growth periods, with the most OTUs detected in seedlings (654), followed by the budding stage (381), flowering stage (350), and flocking stage (351). A total of 217 OTUs were common to all four stages. Pantoea tags were more common to the Shihezi region, whereas Erwinia labels were more common to the Hami region. These results suggest that the dynamics of endophytic bacterial communities are affected by plant growth stage. This highlights the relevance of microbial diversity studies in improving our understanding of endophyte communities.

Diversity and space-time dynamics of the bacterial communities in cotton (Gossypium hirsutum) rhizosphere soil.

Rhizosphere bacteria are key determinants of plant health and productivity. In this study, we used PCR-based next-generation sequencing to reveal the diversity and community composition of bacteria in the cotton rhizosphere from samples collected in Xinjiang Province, China. We identified 125 bacterial classes within 49 phyla from these samples. Proteobacteria (33.07% of total sequences), Acidobacteria (19.88%), and Gemmatimonadetes (11.19%) dominated the bacterial community. Marked differences were evident in the α-diversity of rhizosphere bacteria during different cotton plant growth and development stages. The operational taxonomic unit (OTU) numbers were highest in seedling and bud stages and decreased at the flowering and fruit-boll-opening stages. Forty-three OTUs from the Proteobacteria were common to all four periods of cotton development. Proteobacteria were more abundant in the rhizospheres of cotton from southern Xinjiang than from northern Xinjiang, while the opposite trend was observed for Acidobacteria. Gemmatimonadetes frequency was broadly the same in both northern and southern Xinjiang. These results suggest that there is abundant diversity in the microbiota of cotton rhizosphere soil. Proteobacteria and Actinobacteria dominated this microbial niche and bacterial communities in the seedling, bud, flowering, and boll-opening stages appear to be more similar to one another than to communities at the other growth stages.

Fabry-Pérot Oscillation and Room Temperature Lasing in Perovskite Cube-Corner Pyramid Cavities.

Recently, organometal halide perovskite-based optoelectronics, particularly lasers, have attracted intensive attentions because of its outstanding spectral coherence, low threshold, and wideband tunability. In this work, high-quality CH3 NH3 PbBr3 single crystals with a unique shape of cube-corner pyramids are synthesized on mica substrates using chemical vapor deposition method. These micropyramids naturally form cube-corner cavities, which are eminent candidates for small-sized resonators and retroreflectors. The as-grown perovskites show strong emission ≈530 nm in the vertical direction at room temperature. A special Fabry-Pérot (F-P) mode is employed to interpret the light confinement in the cavity. Lasing from the perovskite pyramids is observed from 80 to 200 K, with threshold ranging from ≈92 µJ cm-2 to 2.2 mJ cm-2 , yielding a characteristic temperature of T0 = 35 K. By coating a thin layer of Ag film, the threshold is reduced from ≈92 to 26 µJ cm-2 , which is accompanied by room temperature lasing with a threshold of ≈75 µJ cm-2 . This work advocates the prospect of shape-engineered perovskite crystals toward developing micro-sized optoelectronic devices and potentially investigating light-matter coupling in quantum optics.