Effects of antimony stress on growth, structure, enzyme activity and metabolism of Nipponbare rice (Oryza sativa L.) roots.

Some antimony (Sb) contaminated areas are used for rice cultivation in response to economic demands. However, little is known about the effects of Sb stress on the growth and metabolism of rice roots. Thus, a hydroponic experiment was carried out on the growth, root anatomy, enzyme activity, and metabolism of Nipponbare rice (Oryza sativa L. ssp. japonica cv. Nipponbare) under varying levels of Sb (III) stress (0 mg L-1, 10 mg L-1, and 50 mg L-1). With the increase of Sb concentration, rice root length and root fresh weight declined by 67.8 % and 90.5 % for 10 mg L-1 Sb stress and 94.1 % and 98.4 % for 50 mg L-1 Sb stress, respectively. Anatomical analysis of cross-sections of Sb-treated roots showed an increase in cell wall thickness and an increase in the number of cell mitochondria. The 10 mg L-1 and 50 mg L-1 Sb stress increased the activity of enzyme superoxide dismutase (SOD) in root cells by 1.94 and 2.40 times, respectively. Compared to the control, 10 mg L-1 Sb treatment increased the activity of catalase (CAT) and peroxidase (POD), as well as the concentrations of antioxidant glutathione (GSH) in the root by 1.46, 1.38, and 0.52 times, respectively. However, 50 mg L-1 Sb treatment significantly decreased the activity or content of CAT, POD and GSH by 28.1 %, 13.5 % and 28.2 %, respectively. Nontargeted LC/MS-based metabolomics analysis identified 23 and 13 significantly differential metabolites in rice roots exposed to 10 mg L-1 and 50 mg L-1 Sb, respectively, compared to the control. These differential metabolites were involved in four main metabolic pathways including the tricarboxylic acid cycle (TCA cycle), butanoate metabolism, alanine, aspartate and glutamate metabolism, and alpha-linolenic acid metabolism. Taken together, these findings indicate that Sb stress destroys the structure of rice roots, changes the activity of enzymes, and affects the metabolic pathway, thereby reducing the growth of rice roots and leading to toxicity.

Diversity and composition of soil bacteria between abandoned and selective-farming farmlands in an antimony mining area.

Background and aims: Land abandonment and selective farming are two common management methods to restore the soil conditions of low-pollution farmland in mining areas. The soil bacterial community plays an important role in farmland soil restoration; however, few studies have compared the composition and diversity of soil bacteria between the abandoned farmlands (AFS) and selective-farming farmlands (FFS). Here, the effects of AFS and FFS on soil properties and bacterial diversity were evaluated in an antimony (Sb) mining area in southern China. This study aimed to identify effective land management methods in terms of positive or negative changes in soil environment and bacterial diversity. Methods: 16S rRNA high-throughput sequencing was used to compare the diversity and composition of soil bacteria between AFS and FFS in the Xikuangshan (the largest Sb mine in the world). Results: Compared to AFS, FFS had higher Sb concentration and nutritional properties (e.g., available N, P, and K) and lower Zn concentration (p < 0.05). The bacterial alpha diversity including Chao1 index, Simpson index, Shannon index and Pielou-e index in FFS was higher than AFS (p < 0.05). At the phylum level, FFS had higher relative abundances of Chloroflexi, Acidobacteria, Gemmatimonadetes, and Rokubacteria, and lower relative abundances of Firmicutes, Actinobacteria, and Bacteroidetes. At the genus level, FFS had higher relative abundances of Acidothermus, Haliangium, and Rokubacteriales, and lower relative abundances of Bacillus, Rhodococcus, Sphingomonas, and 67-14. Redundancy analysis indicated that soil heavy metal content and soil fertility were closely correlated with the soil bacterial community. Altogether, selective farming of low-pollution farmland in the mining area can improve soil properties and soil bacterial diversity.

Assessment of rhizosphere bacterial diversity and composition in a metal hyperaccumulator (Boehmeria nivea) and a nonaccumulator (Artemisia annua) in an antimony mine.

AIMS: Heavy metal hyperaccumulators are widely used in mining restoration due to their ability to accumulate and transport heavy metals, compared to nonaccumulators. Rhizosphere bacteria in metal hyperaccumulators play a key role in the uptake of heavy metals from soil; however, assessments of the differences of rhizosphere bacteria between metal hyperaccumulators and nonaccumulator are scarce. METHODS AND RESULTS: To understand the difference of bacterial composition between hyperaccumulator and nonaccumulator in rhizosphere, the diversity and composition of rhizosphere bacteria in a metal hyperaccumulator (Boehmeria nivea) and a nonaccumulator (Artemisia annua) grown in the same field in Xikuangshan were evaluated using Illumina Miseq high-throughput sequencing technology. Boehmeria nivea and A. annua had 3926 overlapping OTUs, 19,736 and 17,579 unique OTUs, respectively. Boehmeria nivea had lower Chao1 index, Shannon index and Pielou index than A. annua. The dominant phyla and genera of rhizosphere bacteria in B. nivea and A. annua were similar, but some rhizosphere bacterial communities with heavy metal remediation ability mainly appeared in the rhizosphere of the hyperaccumulator. Compared to A. annua, B. nivea showed a significantly higher relative abundance of rhizosphere bacteria, such as Acidobacteria and Bacteroidete at the phylum level and RB41 at the genus level. Some specific rhizosphere bacteria with the ability to bind metal, such as Leifsonia and Kibdelosporangium, were only found in the rhizosphere of B. nivea. CONCLUSION: Results indicated that B. nivea, as a metal hyperaccumulator, has a key function in governing metal-resistant rhizosphere bacteria in response to antimony compound pollution stress. SIGNIFICANCE AND IMPACT OF STUDY: Understanding the diversity of rhizosphere bacteria between hyperaccumulators and nonaccumulators is beneficial to formulate strategies to improve metal uptake efficiency by selecting specific plant species and rhizosphere bacteria grown on polluted soil.

Changes in diversity and composition of rhizosphere bacterial community during natural restoration stages in antimony mine.

BACKGROUND: Open pit antimony (Sb) mining causes serious soil pollution, and phytoremediation is a low-cost approach to remediate heavy metal contaminated soil. Rhizosphere bacteria play an important role in ecological restoration in mining areas. There is a knowledge gap on how to find suitable rhizosphere microorganisms to improve the phytoremediation effect. Understanding the differences of rhizosphere bacterial diversity in different restoration stages is helpful to find suitable bacteria for ecological restoration. METHODS: A method of the substitution of "space" for "time" was used to study the effect of natural restoration on rhizosphere bacterial community. According to the dominant vegetation types (herb, shrub, and tree) in the natural restoration area of Sb mining, the early restoration (ER), middle restoration (MR), and later restoration (LR) from the largest Sb mine (Xikuangshan mine) in the world were selected to evaluate the differences in the composition and diversity of rhizosphere bacteria during three natural restoration stages. Each restoration stage had five samples. To determine the relationship between restoration stages and bacterial diversity in the rhizosphere, high throughput sequencing of PCR amplified were used. RESULTS: Alpha diversity, as assessed by Chao indices, appeared lowest in ER but this trend was not seen with other diversity metrics, including the Simpson and Shannon. Beta diversity analysis suggested there were differences in rhizobacterial community structure associate with restoration stage. At the phylum level, natural restoration led to a significant increase in the relative abundance of Actinobacteria in the MR, and a significant decrease in the relative abundance of Patescibacteria in the LR. Additionally, Calditrichaeota, Deferribacteres and Epsilonbacteraeota were only found in ER. At the genus level, the relative abundance of RB41 and Haliangium were highest in LR plots, while that of Bacillus and Gaiella were highest in ER plots. Additionally, the Azorhizobium genus was only detected in the ER phase. Overall, our findings suggested that several rhizosphere microbial communities had significant differences among three natural restoration stages (ER, MR, and LR) and the rhizosphere bacterial communities mainly appeared in the early restoration stage can be preferred for remediation of pollution soil in Xikuangshan.