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Biology subjects

Santos-Medellin, C.

Publications and source records attributed to Santos-Medellin, C..

13 recordsLinked to original sources

Locally heterogeneous soil viral and prokaryotic responses to prescribed burn correspond with patchy burn severity in a mixed conifer forest

Prescribed burning, a strategy to mitigate wildfires, imparts physicochemical and biological changes to soil. The effects of burns on soil viruses and virus-host dynamics are largely unexplored, despite known viral and prokaryotic contributions to biogeochemical processes. Using a viromic (<0.2 {micro}m size fraction metagenomic) approach, we assessed how viral communities responded to a spring prescribed burn in a mixed conifer forest and whether soil chemical properties and/or prokaryotic host communities could explain the observed patterns. From 120 soil samples (two per depth at 0-3 and 3-6 cm from four burned and two control plots at five timepoints, two before and three after the burn), 91 viromes and 115 16S rRNA gene amplicon libraries were sequenced. Plot location had the greatest effect on explaining variance in viral communities, over treatment (burned or not), depth, and timepoint. Viral and prokaryotic communities exhibited locally heterogenous responses to the fire, with some burned communities resembling unburned controls. This was attributed to patchy burn severity (defined by soil chemistry). Low viromic DNA yields indicated substantial loss of viral biomass in high-severity locations. The relative abundances of Firmicutes, Actinobacteria, and the viruses predicted to infect them significantly increased along the burn severity gradient, suggesting survival of spore formers and viral infection of these abundant, fire-responsive taxa. Our analyses highlight the importance of a nuanced view of soil community responses to fire, not just to burn overall, but to the specific degree of burn severity experienced by each patch of soil, which differed for nearby soils in the same fire.

ecology↗

Virome responses to heating of a forest soil suggest that most dsDNA viral particles do not persist at 90°C

Many fundamental characteristics of soil viruses remain underexplored, including the effects of high temperatures on viruses and their hosts, as would be encountered under disturbances like wildland fire, prescribed burning, and soil solarization. In this study, we leveraged three data types (DNase-treated viromes, non-DNase-treated viromes, and 16S rRNA gene amplicon sequencing) to measure the responses of soil viral and prokaryotic communities to heating to 30{degrees}C, 60{degrees}C, or 90{degrees}C, in comparison to field and control conditions. We investigated (1) the response of dsDNA viral communities to heating of soils from two horizons (O and A) from the same forest soil profile, (2) the extent to which specific viral taxa could be identified as heat-sensitive or heat-tolerant across replicates and soil horizons, and (3) prokaryotic and virus-host dynamics in response to heating. We found that both viral and prokaryotic communities responded similarly to the treatment variables. Community composition differed most significantly by soil source (O or A horizon). Within both soil horizons, viral and prokaryotic communities clustered into three groups, based on beta-diversity patterns: the ambient community (field, control, and 30{degrees}C samples) and the 60{degrees}C and 90{degrees}C communities. As DNase-treated viromic DNA yields were below detection limits at 90{degrees}C, we infer that most viral capsids were compromised after the 90{degrees}C treatment, indicating a maximum temperature threshold between 60{degrees}C and 90{degrees}C for most viral particles in these soils. We also identified groups of heat-tolerant and heat-sensitive vOTUs across both soil sources. Overall, we found that over 70% of viral populations, like their prokaryotic counterparts, could withstand temperatures as high as 60{degrees}C, with shifts in relative abundance explaining most community compositional differences across heating treatments.

genomics↗

Exploring viral particle, soil, and extraction buffer physicochemical characteristics and their impacts on extractable viral communities

Soil viruses are expected to be pivotal members of soil ecosystems, and recent advances in viral size fraction metagenomic (viromic) approaches have substantially improved our ability to interrogate soil viral ecology. However, the first step of viromics relies on extraction buffers to effectively remove viral particles from the soil matrix for downstream analysis, and viral extraction efficiency at this stage could be affected by the interplay between viral particles, soils, and extraction buffer chemistry. Here, we investigated whether extraction buffer chemistry affected extractable viral community composition measured by viromics from different soil types, for both biological (samples collected 1 meter apart) and technical (subsamples from the same soil homogenate) replicates. We first investigated protein-supplemented phosphate-buffered saline pH (PPBS, pHs 4.5, 5.5, 6.5, and 7.5) on a forest, grassland and wetland soil that exhibited different soil edaphic properties, and then we tested different buffer chemistries (PPBS, Carbonated Buffer, Glycine, and Saline Magnesium) on just the wetland soil. Spatial distance, or where the sample was taken in the field (i.e., biological replicate), was the primary driver of extractable viral community composition across all buffers and soils tested. Differences in viral community composition according to extraction buffer properties were only observed in the grassland technical replicates at PPBS buffer pH 4.5, as well as in both the wetland technical and biological replicates treated with different buffer chemistries, but the effects of buffer chemistry were secondary to spatial distance in all cases where spatial distance was a factor. The lack of buffering capacity in the grassland soil technical replicates likely increased sorption of some viral particles at pH 4.5, but neither protein composition nor isoelectric point (both calculated bioinformatically) explained this phenomenon. Given that most soil viral ecological studies to date include sample collection schemes over distances much farther apart than the 1-meter distances considered here, results suggest that extraction buffer chemistry is likely of much lower importance than ecological considerations, such as spatial distance, in the design of future soil viral ecological investigations. HIGHLIGHTSO_LISpatial distance was the main driver of extractable viral community composition. C_LIO_LIExtraction buffer chemistry secondarily structured wetland viral communities. C_LIO_LIAt pH 4.5, PPBS buffer likely increased viral sorption in homogenized grassland soil. C_LIO_LIIncreased sorption was not explained by estimated viral protein isoelectric points. C_LI

microbiology↗

Viral but not bacterial community succession is characterized by extreme turnover shortly after rewetting dry soils

As central members of soil trophic networks, viruses have the potential to drive substantial microbial mortality and nutrient turnover. Pinpointing viral contributions to terrestrial ecosystem processes remains a challenge, as temporal dynamics are difficult to unravel in the spatially and physicochemically heterogeneous soil environment. In Mediterranean grasslands, the first rainfall after seasonal drought provides an ecosystem reset, triggering microbial activity during a tractable window for capturing short-term dynamics. Here, we simulated precipitation in microcosms from four distinct, dry grassland soils and generated 144 viromes and 84 metagenomes to characterize viral, prokaryotic, and relic DNA dynamics over 10 days. Vastly different viral communities in each soil followed remarkably similar successional trajectories. Wet-up triggered a significant increase in viral abundance and richness, followed by extensive compositional turnover. While temporal turnover in prokaryotic communities was much less pronounced, differences in the relative abundances of Actinobacteria (enriched in dry soils) and Proteobacteria (enriched in wetted soils) matched those of their predicted phages, indicating viral predation of dominant bacterial taxa. Rewetting also rapidly depleted relic DNA, which subsequently re-accumulated, indicating substantial new microbial mortality in the days after wet-up, particularly of the taxa putatively under phage predation. Production of abundant, diverse viral particles via microbial host cell lysis appears to be a conserved feature of the early response to soil rewetting, and results suggest the potential for Cull-the-Winner dynamics, whereby viruses infect and cull but do not decimate dominant host populations.

microbiology↗

Substantial differences in soil viral community composition within and among four Northern California habitats

Viruses contribute to food web dynamics and nutrient cycles in diverse ecosystems, yet the biogeographical patterns that underlie these viral dynamics are poorly understood, particularly in soil. Here, we identified trends in soil viral community composition in relation to habitat, moisture content, and physical distance. We generated 30 soil viromes from four distinct habitats (wetlands, grasslands, woodlands, and chaparral) by selectively capturing virus-sized particles prior to DNA extraction, and we recovered 3,432 unique viral species (vOTUs). Viral communities differed significantly by soil moisture content, with viral richness generally higher in wet compared to dry soil habitats. However, vOTUs were rarely shared between samples, including replicates <10 m apart, suggesting that soil viruses may not disperse well and that future soil viral community sampling strategies may need to account for extreme community differences over small spatial scales. Of the 19% of vOTUs detected in more than one sample, 93% were from the same habitat and site, suggesting greater viral community similarity in closer proximity and under similar environmental conditions. Within-habitat differences indicate that extensive sampling would be required for rigorous cross-habitat comparisons, and results belie emerging paradigms of higher viral activity in wet soils and soil viral community spatial heterogeneity.

microbiology↗

Spatial turnover of soil viral populations and genotypes overlain by cohesive responses to moisture in grasslands

Although soil viral abundance, diversity, and potential roles in microbial community dynamics and biogeochemical cycling are beginning to be appreciated1-5, little is known about the patterns and drivers of soil viral community composition that underlie their contributions to terrestrial ecology. Here, we analyzed 43 soil viromes from a precipitation manipulation experiment in a Mediterranean grassland in California, USA. We recovered 5,315 viral population sequences (vOTUs), and viral community composition exhibited a highly significant distance-decay relationship within the 18 m long field. This pattern was recapitulated in the microheterogeneity of 130 prevalent vOTUs (detected in >=90% of the viromes), which tended to exhibit significant negative correlations between genomic similarity of their predominant allelic variants and distance. Although spatial turnover was also observed in the bacterial and archaeal communities from the same soils, the signal was dampened relative to the viromes, suggesting differences in assembly drivers at local scales for viruses and their microbial hosts and/or differences in the temporal scales captured by viromes and total DNA. Despite the overwhelming spatial signal, vOTUs responsive to a decrease in soil moisture were significantly enriched in a predicted protein-sharing subnetwork of 326 vOTUs linked to 191 known actinobacteriophages, suggesting a genomically cohesive viral response to soil moisture evocative of environmental filtering, potentially by way of actinobacterial hosts. Overall, soil viral ecological processes appear to be highly constrained in space and tightly coupled to the heterogeneous, dynamic soil environment and thus fundamentally different from those of their well-mixed and more thoroughly studied marine counterparts.

microbiology↗

RNA viral communities are structured by host plant phylogeny in oak and conifer leaves

Wild plants can suffer devastating diseases, experience asymptomatic, persistent infections, and serve as reservoirs for viruses of agricultural crops, yet we have a limited understanding of the natural plant virosphere. To access representatives of locally and globally distinct wild plants and investigate their viral diversity, we extracted and sequenced dsRNA from leaves from 16 healthy oak and conifer trees in the UC Davis Arboretum (Davis, California). From de novo assemblies, we recovered 389 RNA-dependent RNA polymerase (RdRp) gene sequences from 384 putative viral species, and a further 580 putative viral contigs were identified with virus prediction software followed by manual confirmation of virus annotation. Based on similarity to known viruses, most recovered viruses were predicted to infect plants or fungi, with the highest diversity and abundance observed in the Totiviridae and Mitoviridae families. Phyllosphere viral community composition differed significantly by host plant phylogeny, suggesting the potential for host-specific viromes. The phyllosphere viral community of one oak tree differed substantially from other oak viral communities and contained a greater proportion of putative mycoviral sequences, potentially due to the trees more advanced senescence at the time of sampling. These results suggest that oaks and conifers harbor a vast diversity of viruses with as-yet unknown roles in plant health and phyllosphere microbial ecology.

ecology↗

DNase treatment improves viral enrichment in agricultural soil viromes

The small genomes of most viruses make it difficult to fully capture viral diversity in metagenomes dominated by DNA from cellular organisms. Viral size-fraction metagenomics (viromics) protocols facilitate enrichment of viral DNA from environmental samples, and these protocols typically include a DNase treatment of the post-0.2 m viromic fraction to remove contaminating free DNA prior to virion lysis. However, DNase may also remove desirable viral genomic DNA (e.g., contained in virions compromised due to frozen storage or laboratory processing), suggesting that DNase-untreated viromes might be useful in some cases. In order to understand how virome preparation with and without DNase treatment influences the resultant data, here we compared 15 soil viromes (7 DNase-treated, 8 untreated) from 8 samples collected from agricultural fields prior to tomato planting. DNase-treated viromes yielded significantly more assembled viral contigs, contained significantly less non-viral microbial DNA, and recovered more viral populations (vOTUs) through read mapping. However, DNase-treated and untreated viromes were statistically indistinguishable, in terms of ecological patterns across viral communities. Although results suggest that DNase treatment is preferable where possible, in comparison to previously reported total metagenomes from the same samples, both DNase-treated and untreated viromes were significantly enriched in viral signatures by all metrics compared, including a ~225 times greater proportion of viral reads in untreated viromes compared to total metagenomes. Thus, even without DNase treatment, viromics was preferable to total metagenomics for capturing viral diversity in these soils, suggesting that preparation of DNase-untreated viromes can be worthwhile when DNase treatment is not possible. ImportanceViromics is becoming an increasingly popular method for characterizing soil viral communities. DNase treatment of the viral size fraction prior to DNA extraction is meant to reduce contaminating free DNA and is a common step within viromics protocols to ensure sequences are of viral origin. However, some samples may not be amendable to DNase treatment due to viral particles being compromised either in storage (i.e. frozen) or during other sample processing. To date, the effect of DNase treatment on the recovery of viruses and downstream ecological interpretations of soil viral communities is not thoroughly understood. This work sheds light on these questions and indicates that while DNase treatment of soil viromes improves recovery of viral populations, this improvement is modest in comparison to the gains made by viromics over total soil metagenomics. Further, DNase treatment may not be necessary to observe the ecological patterns structuring soil viral communities.

microbiology↗

Minnesota peat viromes reveal terrestrial and aquatic niche partitioning for local and global viral populations

BackgroundPeatlands are expected to experience sustained yet fluctuating higher temperatures due to climate change, leading to increased microbial activity and greenhouse gas emissions. Despite mounting evidence for viral contributions to these processes in peatlands underlain with permafrost, little is known about viruses in other peatlands. More generally, soil viral biogeography and its potential drivers are poorly understood at both local and global scales. Here, 87 metagenomes and five viral size-fraction metagenomes (viromes) from a boreal peatland in northern Minnesota (the SPRUCE whole-ecosystem warming experiment and surrounding bog) were analyzed for dsDNA viral community ecological patterns, and the recovered viral populations (vOTUs) were compared to our curated PIGEON database of 266,805 vOTUs from diverse ecosystems. ResultsWithin the SPRUCE experiment, viral community composition was significantly correlated with peat depth, water content, and carbon chemistry, including CH4 and CO2 concentrations, but not with temperature during the first two years of warming treatments. Peat vOTUs with aquatic-like signatures (shared predicted protein content with marine and/or freshwater vOTUs) were significantly enriched in more waterlogged surface peat depths. Predicted host ranges for SPRUCE vOTUs were relatively narrow, generally within a single bacterial genus. Of the 4,326 SPRUCE vOTUs, 164 were previously detected in other soils, mostly peatlands. None of the previously identified 202,372 marine and freshwater vOTUs in our PIGEON database were detected in SPRUCE peat, but 1.9% of 78,203 genus-level viral clusters (VCs) were shared between soil and aquatic environments. On a per-sample basis, vOTU recovery was 32 times higher from viromes compared to total metagenomes. ConclusionsResults suggest strong viral "species" boundaries between terrestrial and aquatic ecosystems and to some extent between peat and other soils, with differences less pronounced at the "genus" level. The significant enrichment of aquatic-like vOTUs in more waterlogged peat suggests that viruses may also exhibit niche partitioning on more local scales. These patterns are presumably driven in part by host ecology, consistent with the predicted narrow host ranges. Although more samples and increased sequencing depth improved vOTU recovery from total metagenomes, the substantially higher per-sample vOTU recovery after viral particle enrichment highlights the utility of soil viromics.

microbiology↗

Drought duration determines the recovery dynamics of rice root microbiomes

As extreme droughts become more frequent, dissecting the responses of root-associated microbiomes to drying-wetting events is essential to understand their influence on plant performance. Here, we show that rhizosphere and endosphere communities associated with drought-stressed rice plants display compartment-specific recovery trends. Rhizosphere microorganisms were mostly affected during the stress period, whereas endosphere microorganisms remained altered even after irrigation was resumed. The duration of drought stress determined the stability of these changes, with more prolonged droughts leading to decreased microbiome resilience. Drought stress was also linked to a permanent delay in the temporal development of root microbiomes, mainly driven by a disruption of late colonization dynamics. Furthermore, a root-growth-promoting Streptomyces became the most abundant community member in the endosphere during drought and early recovery. Collectively, these results reveal that severe drought results in enduring impacts on root-associated microbiomes that could potentially reshape the recovery response of rice plants.

microbiology↗

Rice embryogenic trigger BABY BOOM1 promotes somatic embryogenesis by upregulation of auxin biosynthesis genes

Somatic embryogenesis, a powerful tool for clonal propagation and for plant transformation, involves cellular reprogramming of differentiated somatic cells to acquire pluripotency. Somatic embryogenesis can be induced by treating explants with plant growth regulators. However, several plant species including agronomically important cereal crops remain recalcitrant to dedifferentiation and transformation except from embryonic tissues. Somatic embryogenesis can also be induced by ectopic expression of select embryonic factors, including in cereals by BABY BOOM (BBM) transcription factors. How BBM genes bypass the need for exogenous hormones is not well understood. Here, we investigated downstream targets during induction of somatic embryogenesis in rice by OsBBM1 ((Oryza sativa BABY BOOM1). Transient induction of OsBBM1 led to the upregulation of auxin biosynthesis OsYUCCA genes. Continued induction of OsBBM1 resulted in somatic embryogenesis without the need for exogenous auxins. Genetic mutant analysis of OsBBM1 downstream targets, OsYUCCA6, OsYUCCA7 and OsYUCCA9, show that they are required for normal rice development including root and shoot development. Somatic embryogenic potential of OsYUCCA triple mutants was highly compromised despite the presence of exogenous auxin. Additionally, we show that somatic embryogenesis induction by exogenous auxin in rice requires functional BBM genes. Thus, OsBBM1 mediated cellular reprogramming and somatic embryogenesis likely involves increased localized auxin through direct upregulation of OsYUCCA genes. This study reveals mechanistic details of how somatic embryogenesis is established in differentiated tissues in rice, a monocot model and agronomically important cereal crop, with the potential utility to improve regeneration from tissue culture for recalcitrant plants in future. One-sentence summaryRice BABY BOOM1 induces somatic embryogenesis from differentiated tissues by promoting auxin biosynthesis through direct upregulation of YUCCA genes.

developmental biology↗

Viromes outperform total metagenomes in revealing the spatiotemporal patterns of agricultural soil viral communities

Viruses are abundant yet understudied members of soil environments that influence terrestrial biogeochemical cycles. Here, we characterized the dsDNA viral diversity in biochar-amended agricultural soils at the pre-planting and harvesting stages of a tomato growing season via paired total metagenomes and viromes. Size fractionation prior to DNA extraction reduced sources of non-viral DNA in viromes, enabling the recovery of a vaster richness of viral populations (vOTUs), greater viral taxonomic diversity, broader range of predicted hosts, and better access to the rare virosphere, relative to total metagenomes, which tended to recover only the most persistent and abundant vOTUs. Of 2,961 detected vOTUs, 2,684 were recovered exclusively from viromes, while only three were recovered from total metagenomes alone. Both viral and microbial communities differed significantly over time, suggesting a coupled response to rhizosphere recruitment processes and nitrogen amendments. Viral communities alone were also structured along a spatial gradient. Overall, our results highlight the utility of soil viromics and reveal similarities between viral and microbial community dynamics throughout the tomato growing season yet suggest a partial decoupling of the processes driving their spatial distributions, potentially due to differences in dispersal, decay rates, and/or sensitivities to soil heterogeneity.

microbiology↗

Compositional shifts in the root microbiota track the life-cycle of field-grown rice plants

Bacterial communities associated with roots impact the health and nutrition of the host plant. While a multitude of static factors are known to influence the composition of the root-associated microbiota, the dynamics of these microbial assemblies over the plant life cycle are poorly understood. Here, we use dense temporal sampling of spatial compartments to characterize the root-associated microbiota of field grown rice (Oryza sativa) over the course of three consecutive growing seasons and two sites in diverse geographic regions. The root microbiota was found to be highly dynamic during the vegetative phase of plant growth, then stabilizes compositionally for the remainder of the life cycle. Bacterial taxa conserved between field sites can be used as predictive features of rice plant age by modeling using a random forests approach. The age-prediction models were used to reveal that drought stressed plants have developmentally delayed microbiota compared to unstressed plants. Further, by using genotypes with varying developmental rates, we show that shifts in the microbiome are correlated with rates of developmental transitions rather than age alone, such that different microbiota compositions reflect juvenile and adult life stages. These results suggest a model for successional dynamics of the root-associated microbiota over the plant life cycle.

plant biology↗