Biology subjects

Roper, M. C.

Publications and source records attributed to Roper, M. C..

6 recordsLinked to original sources

bioRxiv2026

Orchard management alters citrus root and rhizosphere microbiomes with functional consequences for plant performance

Agricultural management practices act as ecological disturbances that can restructure soil and plant-associated microbial communities, but the functional consequences of these microbial shifts on crop performance remain poorly understood. Here, we examined how common orchard inputs, including wood mulch, glyphosate, and humic acid, affect citrus root and rhizosphere microbiomes and tree performance over a three-year field experiment. Mulch emerged as the dominant driver of microbiome structure, significantly altering bacterial and fungal community composition and increasing rhizosphere alpha diversity. Root microbiomes remained comparatively stable, suggesting stronger host selective forces within root tissues. Mulched rhizospheres were enriched with saprotrophic fungi and metabolically diverse bacteria, while non-mulched soils contained taxa typically associated with nutrient cycling, like Rhizobium, Sphingomonas, and Nitrososphaera. Interactions between mulch and glyphosate further reshaped bacterial communities and corresponded with reduced tree physiological performance, including photosynthesis rates. To verify whether these microbial shifts were contributing to these plant phenotype changes, we conducted a greenhouse experiment using field-derived soil microbiota. Active microbiota from mulch-treated soils reduced citrus seedling establishment and root growth relative to microbiota from non-mulched soils, whereas heat-killed controls eliminated these negative effects, demonstrating a causal relationship between management-induced microbiota changes and decreases in plant performance. In contrast, humic acid influenced plant growth primarily through direct abiotic effects rather than microbial community-level traits. Together, our results show that orchard management practices can restructure citrus microbiomes and generate community-level traits that influence plant performance, highlighting the importance of incorporating microbial ecology and microbiome information when designing and testing crop management strategies.

systems biology↗

bioRxiv2024

In planta transposon sequencing for virulence gene identification in Xylella fastidiosa

In bacterial genetics, large-scale screening approaches such as sequencing transposon mutant pools can be highly effective for identifying and characterizing genes with unknown functions. In the plant pathogen, Xylella fastidiosa, this approach is challenging due to the fastidious nature of this bacterial species and its niche-specific growth in the plant xylem tissue. The purpose of this study was to explore the feasibility of transposon sequencing (Tnseq) for identification of virulence genes in X. fastidiosa, with the hypothesis that this would uncover genes or pathways not previously associated with plant infection. Predicted essential genes were compared after X. fastidiosa strain M23 was grown in vitro and in planta using two known susceptible host species (grapevine and almond). After growth in planta, several gene categories were predicted as essential including hemagglutinins, tRNAs, toxin-antitoxin systems, and prophage genes. Three predicted essential genes (XfasM23_0359, XfasM23_0360, XfasM23_0972) were chosen for further validation by making targeted deletion mutants. Deletion mutants exhibited reduced disease in grapevines, but normal growth and aggregation phenotypes in vitro. Overall, the Tnseq approach has some practical limitations due to the nature of the X. fastidiosa pathosystem, and significant bottleneck effects of inoculation, but was still able to identify genes contributing to disease in plants. Recommendations for future Tnseq studies in X. fastidiosa are discussed based on the challenges and results of this work. ImportanceXylella fastidiosa is a plant pathogenic bacterial species that causes significant economic damage in multiple agricultural industries. Globally, disease epidemics in citrus, grapes, almonds, and olives launched widespread efforts in pathogen surveillance, and quarantine restrictions on plant commodities. Research efforts on X. fastidiosa biology and pathogenesis have still not yielded many new and effective disease control measures, and management in most areas relies primarily on insect vector control. Expansion of available genetic research tools to include high-throughput mutant screening protocols for in planta experiments will facilitate identification of novel disease control targets for this pathogen.

microbiology↗

bioRxiv2023

Permethylation as a Strategy for High Molecular Weight Polysaccharide Structure Analysis by NMR& Case Study of Xylella fastidiosa EPS

Current practices for structure analysis of extremely large molecular weight polysaccharides via solution-state NMR spectroscopy incorporate partial depolymerization protocols that enable polysaccharide solubilization in suitable solvents. Non-specific depolymerization techniques utilized for glycosidic bond cleavage, such as chemical degradation or ultrasonication, potentially generate structure fragments that can complicate the complete characterization of polysaccharide structures. Utilization of appropriate enzymes for polysaccharide degradation, on the other hand, requires prior structure information and optimal enzyme activity conditions that are not available to the analyst working with novel or unknown compounds. Herein, we describe the application of a permethylation strategy that allows the complete dissolution of the intact polysaccharides for NMR structure characterization. This approach is utilized for NMR analysis of Xylella fastidiosa EPS, which is essential for the virulence the plant pathogen that affects multiple commercial crops and is responsible for multibillion dollar losses each year.

biochemistry↗

bioRxiv2022

Using genomes and evolutionary analyses to screen for host-specificity and arms-race dynamics in the plant pathogen Xylella fastidiosa

Xylella fastidiosa infects several economically important crops in the Americas, and it also recently emerged in Europe. Here, using a set of Xylella genomes reflective of the genus-wide diversity, we performed a pan-genome analysis based on both core and accessory genes, for two purposes: i) to test associations between genetic divergence and plant host species and ii) to identify positively selected genes that are potentially involved in arms-race dynamics. For the former, tests yielded significant evidence for specialization of X. fastidiosa to plant host species. This observation contributes to a growing literature suggesting that the phylogenetic history of X. fastidiosa lineages affects host range. For the latter, our analyses uncovered evidence of positive selection across codons for 5.3% (67 of 1,257) of core genes and 5.4% (201 of 3,691) of accessory genes; these genes are candidates to encode interacting factors with plant and insect hosts. Most of these genes had unknown functions, but we identified some tractable candidates including nagZ_2, which encodes a beta-glucosidase that is important for Neisseria gonorrhoeae biofilm formation; cya, which modulates gene expression in pathogenic bacteria; and barA, a membrane associated histidine kinase that has roles in cell division, metabolism, and pili formation. ABSTRACT IMPORTANCEXylella fastidiosa causes devasting diseases to several critical crops. Because X. fastidiosa colonizes and infects many plant species, it is important to understand whether the genome of X. fastidiosa has genetic determinants that underlie specialization to specific host plants. We analyzed genome sequences of X. fastidiosa to investigate evolutionary relationships and to test for evidence of positive selection on specific genes. We found a significant signal between genome diversity and host plants, consistent with bacterial specialization to specific plant hosts. By screening for positive selection, we identified both core and accessory genes that may affect pathogenicity, including genes involved in biofilm formation.

microbiology↗

bioRxiv2022

Arbuscular mycorrhizal fungal composition across US citrus orchards, management strategies, and disease severity spectrum

Arbuscular mycorrhizal fungi (AMF) remain understudied in perennial cropping systems. Citrus is a globally grown fruit tree and under threat by the pandemic Huanglongbing (HLB) disease. Here, we assessed in what capacity geographical location, management strategies and disease affect AMF citrus root communities. Root samples were collected from 88 trees in ten orchards located in the two major citrus producing states in the US. Orchards were selected based on conventional or organic practices in California and based on HLB symptom severity in Florida. We used AMF-specific amplicon sequencing primers to capture community composition and diversity. Taxa names were assigned based on a phylogenetic analysis that comprised a backbone of AMF references sequences from Mycobank and virtual taxa from the MaarjAM database. AMF were detected in 78% of citrus root samples with taxa belonging to six known (Dominikia, Funneliformis, Glomus, Rhizophagus, Sclerocystis, Septoglomus) and unknown Glomeraceae genera. Geographical location affected AMF community composition but not richness, whereas management practice and disease influenced both richness and composition. Our approach indicated that perennial agroecosystems share a set of AMF generalist and specialist taxa. Some taxa could improve environmental fitness and be exploited for agricultural purposes.

microbiology↗

bioRxiv2022

Microbial turnover and dispersal events occur in sync with plant phenology in the perennial evergreen tree crop, Citrus sinensis

Emerging research indicates that plant-associated microbes can alter plant developmental timing. However, it is unclear if host phenology impacts microbial community assembly. Microbiome studies in annuals or deciduous perennial plants face challenges in separating effects of tissue age from phenological driven effects on the microbiome. In contrast, evergreen perennial trees, like Citrus sinensis, retain leaves for years allowing for uniform sampling of similarly aged leaves from the same developmental cohort. This aids in separating phenological effects on the microbiome from impacts due to annual leaf maturation/senescence. Here we used this system to test the hypothesis that host phenology acts as a driver of microbiome composition. Citrus sinensis leaves and roots were sampled during seven phenological stages. Using amplicon-based sequencing, followed by diversity, phylogenetic, differential abundance, and network analyses we examined changes in bacterial and fungal communities. Host phenological stage is the main determinant of microbiome composition, particularly within the foliar bacteriome. Microbial enrichment/depletion patterns suggest that microbial turnover and dispersal were driving these shifts. Moreover, a subset of community shifts were phylogenetically conserved across bacterial clades suggesting that inherited traits contribute to microbe-microbe and/or plant-microbe interactions during specific phenophases. Plant phenology influences microbial community composition. These findings enhance understanding of microbiome assembly and identify microbes that potentially influence plant development and reproduction.

microbiology↗