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Biology subjects

Law, T. F.

Publications and source records attributed to Law, T. F..

5 recordsLinked to original sources

Studying Effects of PDA Media Strengths on the Growth of Colletotrichum sublineola Using MPLEx-Based Integrative Proteomics and Metabolomics Analyses

Colletotrichum sublineola (Cs), the hemibiotrophic fungus that causes sorghum anthracnose, impacts sorghum grain and biomass crop production worldwide. Although nutrient availability is known to influence development in filamentous fungi, including Colletotrichum species, how in vitro nutrient limitation reprograms the Cs cellular state remains unclear. We cultured Cs on full-strength, half-strength, and one-tenth-strength potato dextrose agar (PDA) to define responses across a nutrient gradient. Nutrient limitation induced a pronounced high-sporulation phenotype, with one-tenth-strength PDA producing the strongest conidiation response, followed by half-strength PDA. To study the underlying molecular programs in each condition, we employed a multiplexed metabolite, protein, and lipid extraction (MPLEx) protocol for global proteomics and metabolomics. Global proteomics resulted in 4,590 protein identifications, including 204 unique to one-tenth-strength PDA. Among them are proteins linked to sporulation, vesicular transport, glycosylphosphatidylinositol (GPI)-anchor biosynthesis, and common in fungal extracellular membrane (CFEM)-domain proteins. Differential abundance and pathway analyses revealed a broad reduction of central carbon and energy metabolism, including glycolysis/gluconeogenesis, pentose phosphate, pyruvate metabolism, and glyoxylate pathways, together with increased ribosome-related processes, cAMP signaling, and cell-surface remodeling in one-tenth-strength PDA conditions. In addition, correlative metabolomics supported selective metabolic depletion and resource reallocation toward stress adaptation, membrane remodeling, and conidiation, supporting proteomics findings. Together, these data support a starvation-adapted Cs developmental state associated with enhanced sporulation, cellular pathway reprogramming, and potential virulence linked preparedness under nutrient-limited growth conditions in vitro. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=114 SRC="FIGDIR/small/724728v2_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@f9039borg.highwire.dtl.DTLVardef@5bef0eorg.highwire.dtl.DTLVardef@1abb7b6org.highwire.dtl.DTLVardef@aa11a9_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

Specialization of independently acquired flagellar FliC proteins in plant-associated Sphingomonas balances swimming and immunogenicity

Plants monitor their environment for microbial invaders using pattern-recognition receptors that detect microbe-associated molecular patterns (MAMPs). Flagellin, the main component of bacterial flagellum, contains the flg22 epitope recognized by the plant immune receptor FLS2. Immune recognition can create an evolutionary conflict, requiring bacteria to balance flagellar function and immune evasion. Here, we show that plant-associated Sphingomonads resolve this constraint by partitioning two flagellar functions, motility and colonization, across two divergent and independently acquired flagellin genes. Comparative genomics revealed widespread coexistence of FliC proteins expressing either an immunogenic variant (FliC-H) or a non-immunogenic variant (FliC-L). The non-immunogenic FliC-L is necessary and sufficient for full directional swimming, whereas FliC-H is dispensable for swimming, but sufficient for full attachment and colonization. Flagellin expression patterns mirror these functions. Thus, FLS2 recognizes the flagellar variant required for colonization rather than motility, potentially restricting colonizing bacteria from entering internal leaf and root tissues.

plant biology↗

An efflux pump family distributed across plant commensal bacteria conditions host- and organ-specific detoxfication of a host-specific glucosinolate

In nature, plants recruit a diverse microbial community, the plant microbiome, that is distinct from the surrounding soil community 1-4. To understand the forces that shape the plant microbiome we need to characterize the microbial traits that contribute to plant colonization. We used barcoded mutant libraries to identify bacterial genes that contribute to the colonization of a monocot and a eudicot host 5-7. We show that plant colonization is influenced by dozens of genes. While some of these colonization genes were shared between the two host plant species, most were highly specific, benefiting the colonization of a single host and organ. We characterized an efflux pump that specifically contributes to Arabidopsis shoot colonization. This efflux pump is prevalent across Pseudomonadota genomes, yet benefits the bacterial association with only a small subset of Arabidopsis thaliana accessions. Leveraging genomic diversity within Arabidopsis thaliana, we confirmed that specific glucosinolate breakdown products are detoxified by this family of efflux pumps. The broad prevalence of this efflux pump family suggests that its members contribute to protection of commensal bacteria from collateral damage of plant glucosinolate-based defense responses to herbivores and necrotrophic pathogens.

microbiology↗

Root development is maintained by specific bacteria-bacteria interactions within a complex microbiome

Plants grow within a complex web of species interacting with each other and with the plant. Many of these interactions are governed by a wide repertoire of chemical signals, and the resulting chemical landscape of the rhizosphere can strongly affect root health and development. To understand how microbe-microbe interactions influence root development in Arabidopsis, we established a model system for plant-microbe-microbe-environment interactions. We inoculated seedlings with a 185-member bacterial synthetic community (SynCom), manipulated the abiotic environment, and measured bacterial colonization of the plant. This enabled classification of the SynCom into four modules of co-occurring strains. We deconstructed the SynCom based on these modules, identifying microbe-microbe interactions that determine root phenotypes. These interactions primarily involve a single bacterial genus, Variovorax, which completely reverts severe root growth inhibition (RGI) induced by a wide diversity of bacterial strains as well as by the entire 185-member community. We demonstrate that Variovorax manipulate plant hormone levels to balance this ecologically realistic root communitys effects on root development. We identify a novel auxin degradation operon in the Variovorax genome that is necessary and sufficient for RGI reversion. Therefore, metabolic signal interference shapes bacteria-plant communication networks and is essential for maintaining the roots developmental program. Optimizing the feedbacks that shape chemical interaction networks in the rhizosphere provides a promising new ecological strategy towards the development of more resilient and productive crops.

microbiology↗

The effects of soil phosphorous content on microbiota are driven by the plant phosphate starvation response

Phosphate starvation response (PSR) in non-mycorrhizal plants comprises transcriptional reprogramming resulting in severe physiological changes to the roots and shoots and repression of plant immunity. Thus, plant-colonizing microorganisms - the plant microbiota - are exposed to direct influence by the soils phosphorous (P) content itself, as well as to the indirect effects of soil P on the microbial niches shaped by the plant. The individual contribution of these factors to plant microbiota assembly remains unknown. To disentangle these direct and indirect effects, we planted PSR-deficient Arabidopsis mutants in a long-term managed soil P gradient, and compared the composition of their shoot and root microbiota to wild type plants across different P concentrations. PSR-deficiency had a larger effect on the composition of both bacterial and fungal plant-associated microbiota composition than P concentrations in both roots and shoots. The fungal microbiota was more sensitive to P concentrations per se than bacteria, and less depended on the soil community composition.\n\nUsing a 185-member bacterial synthetic community (SynCom) across a wide P concentration gradient in an agar matrix, we demonstrated a shift in the effect of bacteria on the plant from a neutral or positive interaction to a negative one, as measured by rosette size. This phenotypic shift is accompanied by changes in microbiota composition: the genus Burkholderia is specifically enriched in plant tissue under P starvation. Through a community drop-out experiment, we demonstrate that in the absence of Burkholderia from the SynCom, plant shoots accumulate higher phosphate levels than shoots colonized with the full SynCom, only under P starvation, but not under P-replete conditions. Therefore, P-stressed plants allow colonization by latent opportunistic competitors found within their microbiome, thus exacerbating the plants P starvation.

microbiology↗