bioRxiv Science⌕ Search

Biology subjects

Samad, A.

Publications and source records attributed to Samad, A..

5 recordsLinked to original sources

Differential drought sensitivity of total and active wheat rhizosphere microbiome during rainfall reduction

Root-associated microorganisms play a pivotal role in helping plants adapt to drought stress. However, the underlying mechanisms of the rhizospheric microbiome under limiting soil moisture remain largely unresolved. Integrating total and active microbiome analyses enables a more accurate interpretation of microbial responses to climate change-associated water stress. We assessed the effect of reduced rainfall on two wheat genotypes, drought-tolerant (DT) and drought-sensitive (DS), using rainout shelters that allowed 100%, 75%, 50%, and 25% of natural precipitation to reach the crop. At the peak of the growing season, rhizosphere samples were collected for metagenomic (MG) and metatranscriptome (MT) sequencing. In parallel, rhizosphere volatile organic compounds (VOCs) were collected and analysed. Differential expression analysis of metatranscriptomic data using metagenomic abundance as a cofactor was performed by comparing all treatments to the 100% precipitation control. Our results demonstrate that particularly oxidative stress-related transcripts intensify in DS as rainfall decreases. Transcriptomic shifts primarily involved upregulation of transcripts associated with antioxidant (catalase, superoxide dismutase), heat shock proteins (Hsp10, Hsp60, DnaK/DnaJ, GroEL, GroES), as well as microbial functions related to osmoregulation, proline and glycine betaine (PutA, PutP, OpuBB), and plant growth-promoting traits such as auxin production, phosphate solubilization. Moreover, volatile organic compound (VOC) emissions differed significantly between the control and drought treatments, with higher emissions, particularly acetates, in the DS genotype than in the DT genotype. Overall, pronounced drought-induced shifts in active microbial functions and VOC emissions indicate high sensitivity and functional plasticity of the active microbiome, whereas the total microbiome remains robust under medium drought.

microbiology↗

C-terminal domain of the filamentous hemagglutinin FhaB is crucial for interaction of Bordetella pertussis with ciliated epithelial cells

Bordetella pertussis, the causative agent of whooping cough, produces a [~]370 kDa filamentous hemagglutinin FhaB that serves as a major bacterial adhesin in airway infection. FhaB is secreted via a two-partner secretion pathway and under in vitro culture conditions it is proteolytically processed to the shed [~]230 kDa FHA antigen used currently in acellular pertussis vaccines. We show that FhaB remains largely unprocessed during B. pertussis adhesion to ciliated airway epithelial cells and that its C-terminal domain (CT) is essential for the adhesin function of FhaB. CT deletion did not affect FhaB folding, secretion, or surface exposure, but abolished B. pertussis adhesion to primary human nasal ciliated epithelial cells, thus preventing bacterial colonization of the nasal mucosa and shedding and transmission of the pathogen in a murine nasal infection model. In situ cryo-electron tomography revealed a structural reorganization of the FhaB filaments upon contact with the cilia, presumably due to export of the CT from bacterial periplasm and its subsequent delivery across the ciliary membrane. These findings establish the CT of FhaB as a critical determinant of upper airway colonization by B. pertussis and identify the unprocessed FhaB as the biologically relevant adhesin form involved in airway infection. The revised model of FhaB biogenesis underpins its unique mode of action in pertussis pathogenesis and makes the CT domain to a candidate antigen for future pertussis vaccines.

microbiology↗

Synergistic plant-microbe interactions drive the remediation of naphthenic acid fractional compounds in a constructed wetland mesocosm

Constructed wetland treatment systems (CWTSs) are promising options for treating oil-sands process-affected water (OSPW), which contains toxic naphthenic acid fraction compounds (NAFCs). However, the molecular mechanisms underlying NAFCs attenuation by plants and root microbes remain poorly resolved. In our previous mesocosm study, common cattail (Typha latifolia) increased NAFCs removal by 2.5-fold relative to unplanted controls with no significant effect on plant growth. Here, using RNA from the same experimental system, we applied metatranscriptomics to 40 root samples collected over 60 days to examine plant and active-microbial responses to OSPW exposure. The active-root-associated-microbial community was dominated by Pseudomonadota, which showed a slight increase with exposure to OSPW. Burkholderiales were the most active family, though their relative activity decreased in OSPW systems, where Flavobacteriaceae (Bacteroidota) activity increased. Clear microbial-community shifts were driven by time and OSPW exposure. Although 18 previously proposed microbial NAFC-degradation genes were not differentially expressed, 42 other genes with potential roles in NAFC or related organic compound degradation showed differential expression in OSPW-filled mesocosms. This activity was dominated by specific oxidoreductases from Burkholderiales and Rhizobiales. Crucially, host plant actively responded to OSPW, robustly up-regulating genes encoding oxidoreductases, transporters, and glycosyltransferases, some of which are related to xenobiotic stress and detoxification. Taken together, these results show coordinated plant and microbial transcriptional responses in a system where NAFC removal had already been measured chemically. They help explain the response of OSPW-exposed mesocosms, but the observed patterns likely reflect the broader OSPW mixture rather than NAFCs alone.

ecology↗

Bacteria deliver a microtubule-binding protein into mammalian cells to promote colonization

Pathogenic Bordetella bacteria infect the ciliated respiratory epithelia of mammalian and avian hosts. Several bacterial proteins mediate host cell adhesion, but filamentous hemagglutinin (FhaB) is a principal adhesin because mutants lacking this protein exhibit profound colonization defects. Here, we show that FhaB carries a C-terminal microtubule-binding domain (FhaB-CT), which is translocated into the host-cell cytoplasm to promote bacterial colonization. Cryogenic electron microscopy of microtubule-bound FhaB-CT shows that the domain binds primarily to -tubulin through a network of polar interactions. Live-cell microscopy of infected tracheal explants reveals that FhaB-CT delivery is required for Bordetella to occupy a niche at the base of cilia on airway epithelia. Finally, we demonstrate that the microtubule-binding domain is required for long-term colonization of the mouse nasal cavity by B. pertussis. These observations suggest that the FhaB-CT domain is delivered into motile cilia, where it interacts with axonemal microtubules. We propose that Bordetella initially adhere to the tips of cilia, then deploy multiple FhaB adhesin molecules to migrate to the base of the cilial forest. This mechanism enables Bordetella to resist removal by the mucociliary escalator that clears the respiratory tract of microbes and debris.

microbiology↗

Targeted protein degradation of the CPSF complex by benzoxaboroles through sumoylation

Benzoxaboroles (BoBs) feature a boron-heterocyclic core and are an important innovation in the development of drugs against a range of pathogens and other pathologies. A broad spectrum of pharmacology is associated with chemically diverse BoB derivatives and includes multiple modes-of-action (MoA) and targets. However, a consensus MoA for BoBs targeting evolutionarily diverse protozoan pathogens has emerged with the identification of CPSF3/CPSF73 in the CPSF complex in both apicomplexan and kinetoplastida parasites. Here we establish a functional connection between protein sumoylation and the boron-heterocyclic scaffold shared by all BoBs using comprehensive genetic screens in Trypanosoma brucei. There is a rapid temporal and spatial shift in global protein sumoylation following BoB exposure and members of the CPSF complex are specifically destabilised in a SUMO and proteosome-dependent manner. Finally, we find rapid decrease in bulk mRNA levels, consistent with the role of CPSF3 in mRNA maturation. We propose that a combination of direct inhibition coupled with targeted degradation of CPSF3 underpins the specificity of BoBs against trypanosomatids.

cell biology↗