Rice OsANN9 Enhances Drought Tolerance through Modulating ROS Scavenging Systems.

Drought is a critical abiotic stress which leads to crop yield and a decrease in quality. Annexins belong to a multi-gene family of calcium- and lipid-binding proteins and play diverse roles in plant growth and development. Herein, we report a rice annexin protein, OsANN9, which in addition to regular annexin repeats and type-II Ca2+ binding sites, also consists of a C2H2-type zinc-finger domain. We found that the expression of OsANN9 was upregulated by polyethylene glycol (PEG) or water-deficient treatment. Moreover, plants that overexpressed OsANN9 had increased survival rates under drought stress, while both OsANN9-RNAi and osann9 mutants showed sensitivity to drought. In addition, the overexpression of OsANN9 increased superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities, which regulate reactive oxygen species homeostasis. Collectively, these findings indicate that OsANN9 may function as a positive regulator in response to drought stress by modulating antioxidant accumulation. Interestingly, the setting rates of osann9 mutant rice plants significantly decreased in comparison to wild-type plants, suggesting that OsANN9 might be involved in other molecular mechanisms in the rice seed development stage.

Diversity and abundance of antibiotic resistance genes in rhizosphere soil and endophytes of leafy vegetables: Focusing on the effect of the vegetable species.

Antibiotic resistance genes (ARGs) in the endophytes of vegetables represent a potential route of human exposure to the soil resistome. However, the effect of vegetable species on the endophytic ARG profiles is unclear, hampering our understanding of how ARGs migrate into the soil-vegetable system and their potential health risks. Here, we planted four leafy vegetables (cilantro, endive, lettuce, and pak choi), which are commonly eaten raw, and analyzed the resistomes and microbiomes in three sample types (rhizosphere soil, root, and leaf endophytes). A total of 150 ARG subtypes were detected using high-throughput quantitative PCR. Vegetable species had a significant effect on ARG diversity and abundance, and pak choi accumulated more ARGs in its associated microbiome than the other three vegetables. The bacterial community was the primary factor shaping ARG profiles and was significantly correlated with ARG subtypes. We identified aadE, tet(34), and vanSB as shared ARGs among leaves of the four vegetables; the bacterial families correlated with tet(34) and vanSB were also shared across the vegetables and belonged to Proteobacteria. This study deepens our understanding of how endophytic ARG profiles vary among different vegetables and highlights the potential health risk associated with consuming these vegetables raw.

Effects of Enterococcus faecalis administration on the community structure of airborne bacteria in weanling piglet and layer hen houses.

Probiotics have been shown to improve microbial compositions in animal intestine and feces, but the effects of probiotic administration on airborne microbial composition in animal houses remain unclear. In this study, we investigated the effects of dietary Enterococcus faecalis on the bacterial community structure in the air of piglet and layer hen houses. Indoor air and feces from piglet and layer hen houses were sampled after supplementing E. faecalis in feed for 60 days, and bacterial community structures were analyzed using Illumina high-throughput sequencing technology. Results showed that Chao1, ACE, Shannon, and Simpson indices of bacterial diversity did not significantly change in feces or indoor air of piglet or layer hen after supplementation with E. faecalis (P > 0.05). However, E. faecalis administration resulted in a decrease in the relative abundance of Proteobacteria (P < 0.05). In addition, E. faecalis significantly reduced the relative abundance of opportunistic pathogens such as Acinetobacter, Escherichia, and Shigella (P < 0.05), and beneficial bacterial genus such as Lactobacillus was significantly enriched in both feces and indoor air (P < 0.05). These changes should be of benefit to livestock, farm workers, and the surrounding environment.

[Bacterial Diversity and Community Structure Antibiotic-resistant Bacteria in Bioaerosol of Animal Farms].

Confined animal feeding operations generate high concentrations of airborne antibiotic-resistant bacteria, including pathogenic strains that may pose a health risk to both animals and farm workers and pollute the local air environment. In this study, tetracycline and erythromycin-resistant bacteria were used as examples to study the biodiversity and community structure of airborne antibiotic-resistant bacteria in animal farms. The Anderson sampler was used to collect bioaerosols samples from the inside environment and outside atmospheric environments. A comparative analysis of biological differences of antibiotic-resistant bacteria was conducted on fine and coarse particles, bioaerosol samples inside the house, fecal samples, and inside and outside bioaerosol samples based on the result of the Illumina MiSeq sequencing. The key genus that drives the above differences was also studied. Results showed that the biodiversity of airborne erythromycin-resistant bacteria was higher than that of airborne tetracycline-resistant bacteria, and the biodiversity of bioaerosol samples in the house was higher than that in fecal samples. There were no significant differences in the biodiversity and community structure of airborne antibiotic-resistant bacteria between fine and coarse particles. Actinobacteria is one of the key bacteria responsible for the differences between erythromycin-resistant bacteria and other bacterial populations. Staphylococcus is one of the key genera of tetracycline-resistant flora that is distinguished from erythromycin resistance and all bacterial flora. The results of the community structure showed that there was no significant difference in the dominant flora and the community structure of tetracycline and erythromycin-resistant bacteria. The community structure of feces and bioaerosol samples is different at the genus level, and the dominant bacteria are likewise different. The results of this study provide basic data for the accurate assessment of the current status of antibiotic-resistant bacteria in animal farms and their ecological risks.

Sulfamethoxazole affects the microbial composition and antibiotic resistance gene abundance in soil and accumulates in lettuce.

Pot experiments were set up to simulate the soil contamination by three initial concentrations of sulfamethoxazole (SMX) (S1, 100 mg/kg; S2, 200 mg/kg; S3, 300 mg/kg). The content of SMX in soil and its accumulation in lettuce were analysed. Additionally, the effects of SMX on soil microorganisms and antibiotic resistance genes were studied by Illumina high-throughput sequencing and droplet digital polymerase chain reaction (ddPCR). The results demonstrated that the SMX content in soil reduced by 97%, 86% and 75% in the S1, S2 and S3 treatment groups after 120 days, respectively. The accumulated SMX in lettuce was positively correlated with the initial concentration of SMX in soil. SMX contamination significantly reduced the bacterial diversity and altered the composition of bacterial and fungal communities in soil. The dominant bacterial and fungal genera in the SMX-contaminated soil were obviously different from those in the control soil. The relative abundance of sul1 (sulfonamide resistance gene) remarkably increased in the SMX-contaminated soil, while that of other ARGs, such as sul2 and tetracycline and quinolone resistance genes, showed no significant change.

[Distribution Characteristics of Antibiotic Resistance Genes and Mobile Genetic Elements in Beijing Vegetable Base Soils].

The distribution characteristics of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) in five vegetable base soils from Beijing, China, were assessed. The composition of ARGs and MGEs in soil samples were analyzed by HT-qPCR. We detected 92-121 ARGs and 4-6 MGEs. The ARGs and MGEs in vegetable base soils from different districts were separated from each other. The dominant ARGs shared by vegetable bases were oprD, acrA-04, and acrA-05 of a multidrug, mphA-01 of MLSB, and ß-Lactamase fox5, vanC-03 of vancomycin. The shared MGE among the five vegetable base soils was intI1. A total of seven antibiotics were detected in the soil of several vegetable bases. The dominant antibiotics included enoxacin (ENR), norfloxacin (NOR), oxytetracycline (OTC), and sulfamethoxazole (SMX). The numbers and abundance of antibiotics in the soil of vegetable bases from the Shunyi district were the highest, followed by those from Tongzhou and Changping. Correlation analysis showed that there was a significant positive correlation between the abundance of ARGs and the abundance of antibiotics in the soil of vegetable bases (P<0.05). These results provide basic theoretical data for controlling the transmission of ARGs.

[Microbial Community Structure and the Distribution of Antibiotic Resistance Genes in Soil Contaminated by Sulfamethoxazole].

A pot experiment was carried out to simulate soil contaminated by sulfamethoxazole at different concentrations. The community structure of soil microorganisms was investigated using Illumina high-throughput sequencing, and 64 subtypes of antibiotic resistance genes (ARGs) resistant to six classes of antibiotic were also analyzed by PCR and droplet digital PCR. The results showed that soil contamination with sulfamethoxazole had no significant effect on fungal diversity after 120 days (P>0.05) whereas bacterial diversity was significantly reduced (P<0.05). The microbial community structure of the contaminated soil changed significantly, with the dominant bacterial and fungal genera being significantly different from the control soil. Sulfamethoxazole contamination resulted in an increase in ARG diversity, and the abundance of the sulfonamide resistance gene sul1 increased significantly (P<0.05). However, the abundance of the sulfonamide resistance gene sul2, the quinolone resistance genes floR and cmlA1, and the tetracycline resistance genes tet(34), tetG2, tetG1,tetM, and tetA/P did not show significant changes in the contaminated soil (P>0.05).

Bioaugmentation with Diaphorobacter polyhydroxybutyrativorans to enhance nitrate removal in a poly (3-hydroxybutyrate-co-3-hydroxyvalerate)-supported denitrification reactor.

A newly isolated and identified Diaphorobacter polyhydroxybutyrativorans strain (SL-205) was employed to enhance the denitrification performance of a laboratory-scale solid-phase denitrification (SPD) reactor using poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) as a carbon source, and dynamic variations in microbial communities in the reactor were investigated. Results indicated that bioaugmentation with strain SL-205 enabled rapid reactor startup and improved denitrification performance relative to the reactor inoculated with activated sludge. Illumina sequencing revealed that bioaugmentation also significantly increased Proteobacteria abundance along with increased influent nitrate loading. Additionally, two genera of PHBV-degrading denitrifers, Diaphorobacter and Acidovorax, exhibited higher abundance, and elevated expression of denitrification-associated genes (narG, nirK, and nirS) was observed following bioaugmentation relative to the control at influent nitrate loading ranging from 1.28 g N/(L·d) to 1.6 g N/(L·d).

Sphingomonaszeicaulis sp. nov., an endophytic bacterium isolated from maize root.

A novel Gram-staining-negative, aerobic and rod-shaped strain designated 541T was isolated from surface-sterilized root tissue of maize, collected from the Fangshan District of Beijing, People's Republic of China, and was subjected to a taxonomic study using a polyphasic approach. According to a phylogenetic tree based on 16S rRNA gene sequences, strain 541T represented a member of the genus Sphingomonas and clustered with Sphingomonas sanxanigenens DSM 19645T, with which it shared the highest 16S rRNA gene sequence similarity (98.8 %). The predominant respiratory quinone was ubiquinone-10 (Q-10), the major polyamine was sym-homospermidine and the major cellular fatty acids were C18 : 1ω7c (50.9 %), C16 : 0 (22.0 %) and C14 : 0 2-OH (11.4 %). The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine and sphingoglycolipid. The DNA G+C content was 64.7 mol%. DNA-DNA relatedness between strain 541T and its closest phylogenetic relative Sphingomonas sanxanigenens DSM 19645T was 50.8 %. The results of physiological and biochemical tests and the differences in the fatty acid profiles allowed a clear phenotypic differentiation of strain 541T from closely related species of the genus Sphingomonas. Strain 541T represents a novel species within the genus Sphingomonas, for which the nameSphingomonas zeicaulis sp. nov. is proposed, with the type strain 541T (=CGMCC 1.15008T=DSM 100587T).