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.

Identification of the rhizospheric microbe and metabolites that led by the continuous cropping of ramie (Boehmeria nivea L. Gaud).

Continuous cropping lowers the production and quality of ramie (Boehmeria nivea L. Gaud). This study aimed to reveal the metagenomic and metabolomic changes between the healthy- and obstacle-plant after a long period of continuous cropping. After 10 years of continuous cropping, ramie planted in some portions of the land exhibited weak growth and low yield (Obstacle-group), whereas, ramie planted in the other portion of the land grew healthy (Health-group). We collected rhizosphere soil and root samples from which measurements of soil chemical and plant physiochemical properties were taken. All samples were subjected to non-targeted gas chromatograph-mass spectrometer (GS/MS) metabolome analysis. Further, metagenomics was performed to analyze the functional genes in rhizospheric soil organisms. Based on the findings, ramie in Obstacle-group were characterized by shorter plant height, smaller stem diameter, and lower fiber production than that in Health-group. Besides, the Obstacle-group showed a lower relative abundance of Rhizobiaceae, Lysobacter antibioticus, and Bradyrhizobium japonicum, but a higher relative abundance of Azospirillum lipoferum and A. brasilense compared to the Health-group. Metabolomic analysis results implicated cysteinylglycine (Cys-Gly), uracil, malonate, and glycerol as the key differential metabolites between the Health- and Obstacle-group. Notably, this work revealed that bacteria such as Rhizobia potentially synthesize IAA and are likely to reduce the biotic stress of ramie. L. antibioticus also exerts a positive effect on plants in the fight against biotic stress and is mediated by metabolites including orthophosphate, uracil, and Cys-Gly, which may serve as markers for disease risk. These bacterial effects can play a key role in plant resistance to biotic stress via metabolic and methionine metabolism pathways.

Genome Sequence Resource for the Ramie Oomycete Pathogen Phytopythium vexans HF1.

The oomycete Phytopythium vexans is a causative agent of patch canker, damping-off, and crown, stem, and root rot in many economically important plants. P. vexans HF1 was isolated in China, where it caused brown root rot of ramie, a fiber crop broadly cultivated in Asia. The genome of HF1 was sequenced by a combination of technologies producing short (Illumina HiSeq X) and long (PacBio RS) reads. The genome is 41.73 Mbp long, assembled into 44 contigs. It has a GC content of 58.17% and contains 13,051 predicted coding genes, including 1,461 putative virulence genes and 220 putative antimicrobial resistance genes. This genome sequence provides a resource for determining the molecular mechanisms of disease development in this pathosystem.

Analysis of bacterial and fungal communities in continuous-cropping ramie (Boehmeria nivea L. Gaud) fields in different areas in China.

Ramie (Boehmeria nivea L. Gaud) suffers from long-term continuous cropping. Here, using Illumina high-throughput sequencing technology, we aimed to identify bacteria and fungi associated with continuous cropping in ramie fields in Yuanjiang, Xianning, Sichuan, and Jiangxi. The rarefaction results showed that Jiangxi had significantly lower bacterial α-diversity than that of the other areas. Firmicutes, Proteobacteria, and Acidobacteria were the dominant bacterial phyla, and Ascomycota, Basidiomycota, and Zygomycota were the dominant fungal phyla. In Jiangxi, Firmicutes accounted for 79.03% of all valid reads, which could have significant decreased microbial diversity and negative effects of continuous ramie cropping. We used traditional methods to examine soil nutrients. Sichuan had a relatively high pH and available P and K, but low total N; opposite findings were recorded in Jiangxi. The redundancy analysis revealed that the urease activity, PH, available K, and total N significantly correlated with bacterial community abundance, whereas only total N significantly correlated with fungal community abundance (P < 0.01). Overall, the effect of soil environmental factors on the bacterial diversity of continuous ramie cropping was greater than that on fungal diversity. In the future, we will focus on the effect of rhizosphere bacteria to solve the obstacle in continuous ramie cropping.

Potential use of high-throughput sequencing of soil microbial communities for estimating the adverse effects of continuous cropping on ramie (Boehmeria nivea L. Gaud).

Ramie (Boehmeria nivea L. Gaud) fiber, one of the most important natural fibers, is extracted from stem bark. Continuous cropping is the main obstacle to ramie stem growth and a major cause of reduced yields. Root-associated microbes play crucial roles in plant growth and health. In this study, we investigated differences between microbial communities in the soil of healthy and continuously cropped ramie plants, and sought to identify potential mechanisms whereby these communities could counteract the problems posed by continuous cropping. Paired-end Illumina MiSeq analysis of 16S rRNA and ITS gene amplicons was employed to study bacterial and fungal communities. Long-term monoculture of ramie significantly decreased fiber yields and altered soil microbial communities. Our findings revealed how microbial communities and functional diversity varied according to the planting year and plant health status. Soil bacterial diversity increased with the period of ramie monoculture, whereas no significant differences were observed for fungi. Sequence analyses revealed that Firmicutes, Proteobacteria, and Acidobacteria were the most abundant bacterial phyla. Firmicutes abundance decreased with the period of ramie monoculture and correlated positively with the stem length, stem diameter, and fiber yield. The Actinobacteria, Chloroflexi, and Zygomycota phyla exhibited a significant (P < 0.05) negative correlation with yields during continuous cultivation. Some Actinobacteria members showed reduced microbial diversity, which prevented continuous ramie cropping. Ascomycota, Zygomycota, and Basidiomycota were the main fungal phyla. The relatively high abundance of Bacillus observed in healthy ramie may contribute to disease suppression, thereby promoting ramie growth. In summary, soil weakness and increased disease in ramie plants after long-term continuous cropping can be attributed to changes in soil microbes, a reduction in beneficial microbes, and an accumulation of harmful microbes.

Diversity of Arbuscular Mycorrhizal Fungi Associated with a Sb Accumulator Plant, Ramie (Boehmeria nivea), in an Active Sb Mining.

Arbuscular mycorrhizal fungi (AMF) have great potential for assisting heavy metal hyperaccumulators in the remediation of contaminated soils. However, little information is available about the symbiosis of AMF associated with an antimony (Sb) accumulator plant under natural conditions. Therefore, the objective of this study was to investigate the colonization and molecular diversity of AMF associated with the Sb accumulator ramie (Boehmeria nivea) growing in Sb-contaminated soils. Four Sb mine spoils and one adjacent reference area were selected from Xikuangshan in southern China. PCR-DGGE was used to analyze the AMF community composition in ramie roots. Morphological identification was also used to analyze the species in the rhizosphere soil of ramie. Results obtained showed that mycorrhizal symbiosis was established successfully even in the most heavily polluted sites. From the unpolluted site Ref to the highest polluted site T4, the spore numbers and AMF diversity increased at first and then decreased. Colonization increased consistently with the increasing Sb concentrations in the soil. A total of 14 species were identified by morphological analysis. From the total number of species, 4 (29%) belonged to Glomus, 2 (14%) belonged to Acaulospora, 2 (14%) belonged to Funneliformis, 1 (7%) belonged to Claroideoglomus, 1 (7%) belonged to Gigaspora, 1 (7%) belonged to Paraglomus, 1 (7%) belonging to Rhizophagus, 1 (7%) belonging to Sclervocystis, and 1 (7%) belonged to Scutellospora. Some AMF sequences were present even in the most polluted site. Morphological identification and phylogenetic analysis both revealed that most species were affiliated withGlomus, suggesting that Glomus was the dominant genus in this AMF community. This study demonstrated that ramie associated with AMF may have great potential for remediation of Sb-contaminated soils.

Nitrogen removal of ramie stalk treated by acid wastewater combined with Clostridium thermocellum and the kinetic study of pyrolysis.

A pretreatment was developed to remove nitrogen from ramie residue and cotton stalk to eliminate potential nitrogen pollutants in biomass energy production. In the pretreatment, straw was treated with acid wastewater from bioleaching for 3 h followed by Clostridium thermocellum incubation for 2 h. Most nitrogen was removed from biomass waste and the major was that in protein. Pyrolysis process revealed most hemicellulose was removed and the kinetics fitted the first-order equation well. Apparent activation energy of ramie residue increased a little and mass loss became concentrated. Ultimate analysis and pyrolysis analysis revealed the treatment did not weaken the application value of biomass in energy production. Replacing acid wastewater with sulphuric acid, a higher nitrogen removal could be achieved; however, activation energy increased sharply.

Isolation of high quality RNA and construction of a suppression subtractive hybridization library from ramie (Boehmeria nivea L. Gaud.).

Isolation of high quality RNA from ramie (Boehmeria nivea L. Gaud.) is difficult due to its high levels of polyphenols, polysaccharides, pectin, fat, wax and other secondary metabolites. A modified procedure based on guanidinium isothiocyanate for RNA preparation of ramie was developed in this study. High concentrations (5%, v/v) of guanidinium isothiocyanate, PVP-4000, sodium citrate and sodium lauryl sarcosinate and beta-mercaptoethanol were used in the extraction buffer, together with a low pH sodium acetate (pH 4.0) added to improve the RNA quality. The average yield was about 400 microg RNAg(-1) fresh leaves. One SSH library which was induced by ramie anthracnose was constructed by utilizing the RNA extracted through the present method. These results showed that our protocol was applicable for RNA isolation from recalcitrant ramie tissues.