Highly divergent and diverse viral community infecting sylvatic mosquitoes from Northeast Brazil.

Mosquitoes can transmit several pathogenic viruses to humans, but their natural viral community is also composed of a myriad of other viruses such as insect-specific viruses (ISVs) and those that infect symbiotic microorganisms. Besides a growing number of studies investigating the mosquito virome, the majority are focused on few urban species, and relatively little is known about the virome of sylvatic mosquitoes, particularly in high biodiverse biomes such as the Brazilian biomes. Here, we characterized the RNA virome of 10 sylvatic mosquito species from Atlantic forest remains at a sylvatic-urban interface in Northeast Brazil employing a metatranscriptomic approach. A total of 16 viral families were detected. The phylogenetic reconstructions of 14 viral families revealed that the majority of the sequences are putative ISVs. The phylogenetic positioning and, in most cases, the association with a high RNA-dependent RNA polymerase amino acid divergence from other known viruses suggests that the viruses characterized here represent at least 34 new viral species. Therefore, the sylvatic mosquito viral community is predominantly composed of highly divergent viruses highlighting the limited knowledge we still have about the natural virome of mosquitoes in general. Moreover, we found that none of the viruses recovered were shared between the species investigated, and only one showed high identity to a virus detected in a mosquito sampled in Peru, South America. These findings add further in-depth understanding about the interactions and coevolution between mosquitoes and viruses in natural environments. IMPORTANCE: Mosquitoes are medically important insects as they transmit pathogenic viruses to humans and animals during blood feeding. However, their natural microbiota is also composed of a diverse set of viruses that cause no harm to the insect and other hosts, such as insect-specific viruses. In this study, we characterized the RNA virome of sylvatic mosquitoes from Northeast Brazil using unbiased metatranscriptomic sequencing and in-depth bioinformatic approaches. Our analysis revealed that these mosquitoes species harbor a diverse set of highly divergent viruses, and the majority comprises new viral species. Our findings revealed many new virus lineages characterized for the first time broadening our understanding about the natural interaction between mosquitoes and viruses. Finally, it also provided several complete genomes that warrant further assessment for mosquito and vertebrate host pathogenicity and their potential interference with pathogenic arboviruses.

Metatranscriptome Revealed the Efficacy and Safety of a Prospective Approach for Agricultural Wastewater Reuse: Achieving Ammonia Retention during Biological Treatment by a Novel Natural Inhibitor Epsilon-Poly-l-Lysine.

Appropriate inhibitors might play important roles in achieving ammonia retention in biological wastewater treatment and its reuse in agriculture. In this study, the feasibility of epsilon-poly-l-lysine (ε-PL) as a novel natural ammonia oxidation inhibitor was investigated. Significant inhibition (ammonia oxidation inhibition rate was up to 96.83%) was achieved by treating the sludge with ε-PL (400 mg/L, 12 h soaking) only once and maintaining for six cycles. Meanwhile, the organic matter and nitrite removal was not affected. This method was effective under the common environmental conditions of biological wastewater treatment. Metatranscriptome uncovered the possible action mechanisms of ε-PL. The ammonia oxidation inhibition was due to the co-decrease of Nitrosomonas abundance, ammonia oxidation genes, and the cellular responses of Nitrosomonas. Thauera and Dechloromonas could adapt to ε-PL by stimulating stress responses, which maintained the organic matter and nitrite removal. Importantly, ε-PL did not cause the enhancement of antibiotic resistance genes and virulent factors. Therefore, ε-PL showed a great potential of ammonia retention, which could be applied in the biological treatment of wastewater for agricultural reuse.

Root microbiome changes associated with cadmium exposure and/or overexpression of a transgene that reduces Cd content in rice.

Cadmium (Cd) is a toxic heavy metal that can accumulate in crop plants. We reported previously the engineering of a low cadmium-accumulating line (2B) of rice through overexpression of a truncated OsO3L2 gene. As expression of this transgene was highest in plant roots, amplicon and metatranscriptome sequencing were used to investigate the possibility that its expression affects root associated microbes. Based on amplicon sequencing of bacterial 16S rRNA, but less so from fungal ITS, the OTUs (operational taxonomic units) showed less diversity in soil tightly (rhizoplane) than loosely (rhizosphere) associated with plant roots. Significantly changed OTUs caused by the low-Cd accumulating plant 2B, Cd treatment or both were found, and 10 of the 13 OTUs (77%) that were enriched in Cd treated 2B samples over the wild type counterpart have been previously described as involved in tolerance to Cd or other heavy metals. Metatranscriptome sequencing of rhizosphere microbiome found that bacteria accounted for 70-75% of the microbial RNA. Photosynthesis-antenna proteins and nitrogen metabolism pathways were most active in soil microbes treated with Cd and grown with plant 2B. Correspondingly, the relative abundance of Cyanobacteria was enriched to < 1% of Cd treated rhizosphere bacteria, yet accounted for up to 13% of Cd treated 2B rhizospheric transcripts. These enriched microbes by transgene and Cd are worthy candidates for future application on reducing crop uptake of Cd.

Integrated metatranscriptomics and metabolomics reveal microbial succession and flavor formation mechanisms during the spontaneous fermentation of Laotan Suancai.

Laotan Suancai, a Chinese traditional fermented vegetable, possesses a unique flavor that depends on the fermentative microbiota. However, the drivers of microbial succession and the correlation between flavor and active microbiota remain unclear. A total of 21 characteristic flavor metabolites were identified in Laotan Suancai by metabolomics, including 8 sulfides, 6 terpenes, 3 organic acids, 2 isothiocyanates, 1 ester, and 1 pyrazine. Metatranscriptome analysis revealed variations in the active microbiota at different stages of fermentation, and further analysis indicated that organic acids were the primary drivers of microbial succession. Additionally, we reconstructed the metabolic network responsible for the formation of characteristic flavor compounds and identified Companilactobacillus alimentarius, Weissella cibaria, Lactiplantibacillus plantarum, and Loigolactobacillus coryniformis as the core functional microbes involved in flavor development. This study contributed to profoundly understanding the relationship between the active microbiota and flavor quality formation, as well as the targeted selection of starters with flavor regulation abilities.

Diversity of Mycoviruses Present in Strains of Binucleate Rhizoctonia and Multinucleate Rhizoctonia, Causal Agents for Potato Stem Canker or Black Scurf.

In this study, the diversity of putative mycoviruses present in 66 strains of binucleate Rhizoctonia (BNR, including anastomosis group (AG)-A, AG-Fa, AG-K, and AG-W) and 192 strains of multinucleate Rhizoctonia (MNR, including AG-1-IA, AG-2-1, AG-3 PT, AG-4HGI, AG-4HGII, AG-4HGIII, and AG-5), which are the causal agents of potato stem canker or black scurf, was studied using metatranscriptome sequencing. The number of contigs related to mycoviruses identified from BNR and MNR was 173 and 485, respectively. On average, each strain of BNR accommodated 2.62 putative mycoviruses, while each strain of MNR accommodated 2.53 putative mycoviruses. Putative mycoviruses detected in both BNR and MNR contained positive single-stranded RNA (+ssRNA), double-stranded RNA (dsRNA), and negative single-stranded RNA (-ssRNA) genomes, with +ssRNA genome being the prevalent nucleic acid type (82.08% in BNR and 75.46% in MNR). Except for 3 unclassified, 170 putative mycoviruses found in BNR belonged to 13 families; excluding 33 unclassified, 452 putative mycoviruses found in MNR belonged to 19 families. Through genome organization, multiple alignments, and phylogenetic analyses, 4 new parititviruses, 39 novel mitoviruses, and 4 new hypoviruses with nearly whole genome were detected in the 258 strains of BNR and MNR.