Biology subjects

Mostofa, M. G.

Publications and source records attributed to Mostofa, M. G..

8 recordsLinked to original sources

bioRxiv2026

RAP2.6 negatively regulates Arabidopsis submergence tolerance through GSNOR1-dependent redox and hormonal signaling

Group VII ETHYLENE RESPONSE FACTORs (ERFVIIs) are central regulators of plant responses to oxygen deprivation. However, the contributions of many other APETALA2 (AP2)/ERF transcription factors to hypoxia and flooding responses remain poorly understood. Here, we identify the group X ERF transcription factor RAP2.6 as a negative regulator of submergence tolerance in Arabidopsis thaliana. RAP2.6 expression declined under hypoxia, and plants overexpressing RAP2.6 exhibited reduced survival following dark submergence, whereas the rap2.6-1 loss-of-function mutant showed enhanced tolerance to submergence, hypoxia, and ethanol treatment. Enhanced tolerance of rap2.6-1 was associated with reduced membrane injury, maintenance of photosynthetic pigments and soluble sugars, and extensive metabolic remodeling. RAP2.6 overexpression altered nitric oxide (NO), S-nitrosothiol (SNO), and ethylene accumulation during submergence and was associated with greater oxidative damage and enhanced abscisic acid (ABA)-responsive gene expression. Genetic analyses using S-NITROSOGLUTATHIONE (GSNO) REDUCTASE 1 (GSNOR1) gain- and loss-of-function backgrounds demonstrated that GSNOR1-dependent GSNO homeostasis strongly modified RAP2.6-dependent submergence responses. Increased GSNOR1 activity suppressed the submergence-sensitive phenotype of RAP2.6-overexpressing plants, reduced nitro-oxidative stress, and restored carbohydrate reserves, whereas loss of GSNOR1 activity abolished the enhanced tolerance of rap2.6-1. GSNOR1 also modified RAP2.6-dependent expression of genes associated with oxygen sensing and ABA signaling, while its protective effect was not explained solely by changes in ethylene accumulation. Together, these findings identify RAP2.6 as a regulatory component of the low-oxygen response and reveal a functional interaction between RAP2.6 and GSNOR1-dependent redox signaling that coordinates plant hormonal, metabolic, and oxidative responses during submergence.

plant biology↗

bioRxiv2026

Integrative genetic and structural analyses reveal nitric oxide-driven regulation of ERFVII stability in Arabidopsis under low O2

Nitric oxide (NO) is a key signaling molecule that regulates diverse physiological responses, including adaptation to hypoxia in plants. Although stabilization of group-VII ETHYLENE RESPONSE FACTOR (ERFVII) transcription factors under low O2 is known to be facilitated by the N-dragon pathway, whether NO directly mediates N-terminal cysteine (Cys) oxidation to regulate ERFVII stability remains unresolved. Here, we combined genetic, computational, and structural approaches to investigate NOs role in proteasomal degradation of ERFVII during flooding stress. Arabidopsis mutants with elevated S-Nitrosoglutathione (GSNO) levels compromised ERFVII activation during dark submergence but increased expression of genes encoding N-degron pathway enzymes. Although direct detection of in vivo NO-mediated S-nitrosylated proteins remains technically challenging, the GPS-SNO 1.0 tool predicted the conserved N-terminal second Cys residue as a high-confidence S-nitrosylation site. Structural modeling using the Schrodinger Suite 2024-4 further revealed that conversion of the Cys thiol (Cys-SH) to S-nitrosothiol (Cys-SNO) induced notable conformational changes in ERFVII TFs. Molecular docking further demonstrated that Cys-SNO-modified ERFVII peptides exhibited stronger binding affinities and altered interaction networks with N-degron pathway enzymes, supporting a role for NO-mediated structural remodeling in ERFVII degradation. Elevated GSNO also disrupted energy balance efficiency for overcoming O2 deficiency, altered the expression of sugar starvation-responsive genes, and impaired ATP binding capacity of Cys-SNO ERFVII proteins, as evidenced by docking and molecular dynamics simulations. Collectively, these results support a model in which NO-mediated modification of the conserved N-terminal Cys promotes ERFVII degradation, thereby linking NO signaling to hypoxia-responsive transcriptional regulation and metabolic adaptation during flooding stress.

plant biology↗

bioRxiv2026

AP2/ERF transcription factor RAP2.6 regulates early flowering in Arabidopsis thaliana by altering S-nitrosothiol levels and cytokinin responses

RAP2.6, an AP2/ERF transcription factor (TF), regulates plant stress responses; however, its role in floral transition remains unexplored. Here, we evaluated RAP2.6s role in flowering and the associated transcriptional changes in Arabidopsis thaliana under long-day conditions. RAP2.6-overexpressing line showed early flowering with fewer rosette leaves, whereas rap2.6-1 mutant flowered later, had more rosette leaves, and higher expression of the floral repressor FLOWERING LOCUS C (FLC). Early flowering in the overexpressing line was accompanied by transcriptional activation of the floral integrators GIGANTEA (GI), FLOWERING LOCUS T (FT), and COSTANS (CO), potentially through RAP2.6 interaction with GCC/DRE cis-regulatory elements. RAP2.6-mediated floral transition depended on nitric oxide (NO), with flowering time largely varying based on NO bioactivity. RAP2.6 was found to be a downstream regulator of Arabidopsis S-NITROSOGLUTATHIONE REDUCTASE 1 (GSNOR1) in controlling S-nitrosothiol (SNO) levels, flowering time, and silique formation. The NITRIC OXIDE-ASSOCIATED 1 (NOA1)-dependent reduction in NO levels abolished early flowering in 35S::RAP2.6 plants without affecting silique formation. Furthermore, enhanced cytokinin sensitivity and upregulation of cytokinin biosynthetic genes suggest cytokinin involvement in RAP2.6-mediated flowering. Together, these findings highlight the crucial role of RAP2.6 in regulating flowering time by integrating redox and hormonal signaling to coordinate reproductive development in A. thaliana.

plant biology↗

bioRxiv2025

Bacillus subtilis reprograms host transcriptome and rhizosphere microbiome via systemic signaling to confer alkaline stress tolerance in garden pea

Soil alkalinity severely limits legume growth, but the role of Bacillus subtilis in alkaline stress tolerance remains unclear in garden pea. We found that multiple garden pea genotypes inoculated with B. subtilis under alkaline stress showed host-specific improvements in growth parameters. Mechanistic analysis conducted on Sugar Snap showed improved nodulation, mineral status, and photosystem efficiency, while split-root assays showed responses consistent with systemic effects of B. subtilis in alkaline tolerance. Further, FeEDDHA partially reduced alkaline stress symptoms but did not fully restore nodulation. In contrast, B. subtilis increased rhizosphere Fe-chelating activity and improved nodulation, leading to stronger symbiotic recovery than inorganic Fe alone. This suggests that factors associated with B. subtilis inoculation, beyond Fe availability alone, may contribute to the observed recovery of nodulation. This is further supported by in vitro co-culture experiments showing enhanced growth of R. leguminosarum in the presence of B. subtilis under alkaline conditions, indicating potential microbial compatibility for coping with stress. RNA-seq analysis identified 958 upregulated and 1,134 downregulated genes in roots inoculated with B. subtilis under alkaline conditions. The upregulated genes were mostly involved in the sugar-mediated symbiotic association (SWEET and GLUT), pH homeostasis (cation/H+ exchanger and ATPase), and nutrient assimilation (Ammonium transporter and Zn/Fe permease). Microbial community analysis revealed that B. subtilis significantly altered bacterial alpha diversity under alkaline stress, whereas fungal alpha diversity remained unaffected. Further, B. subtilis reshaped the rhizosphere microbial community and enriched taxa such as Pseudomonas, Pseudorhizobium, and Chaetomium, which were potentially associated with responses to alkaline stress. Taken together, microbial interventions such as B. subtilis offer an effective strategy to boost legume tolerance to alkaline soils.

plant biology↗

bioRxiv2024

Isoprene deters insect herbivory by priming plant hormone responses

Isoprene, emitted by some plants, enhances plant abiotic resilience but its role in biotic stress resilience remains elusive. We used tobacco plants engineered to emit isoprene (IE) and the corresponding azygous non-emitting control (NE) to investigate isoprene emission and biotic stress resilience. IE plants were more resistant to insect herbivory than NE plants. Worms preferred to feed on NE rather than IE leaves. IE plants showed less decline in photosynthesis during worm feeding. Insect feeding increased jasmonate levels in IE leaves, suggesting isoprene-mediated priming of the jasmonic acid response. Wound-induced increase in isoprene emission corresponded with elevation of methyl-D-erythritol-4-phosphate pathway and Calvin-Benson cycle metabolites. The results highlight interactive functions of isoprene and jasmonic acid and advance our understanding of how isoprene emission enhances plant resilience.

plant biology↗

bioRxiv2024

Fe deficiency causes transcriptional shift in roots leading to disruption of drought tolerance in soybean

Soil alkalinity and drought collectively cause severe growth retardation in crops. However, the adaptive responses and transcriptional changes under such conditions remain unclear in soybean. In this growth incubator study, soil alkalinity and drought stress led to significant reductions in plant biomass, chlorophyll, and nutrient uptake in soybean. However, the photochemical efficiency of photosystem II remained stable, suggesting the activation of protective mechanisms to maintain photosynthetic functions. RNA-seq analysis demonstrated 357 upregulated and 799 downregulated genes in roots due to combined soil alkalinity and drought. Analysis revealed a complex response, with upregulation of genes predominantly involved in mineral homeostasis (Iron dehydrogenase, Sulfurylase), reactive oxygen species scavenging (Glutamate synthase, L-Ascorbate peroxidase) and hormonal signaling. Particularly, several ethylene-responsive genes, including the Transcription factors TF5 and TF018, were upregulated, indicating the activation of stress-related signaling pathways. In a targeted study, plants supplemented with an ethylene precursor showed significant improvements in morpho-physiological traits and Fe status under combined stress. However, ethylene precursor applied in non-stressed plants led to reduced growth, and Fe levels, suggesting an involvement of the context-dependent role of ethylene in promoting stress tolerance. Furthermore, ethylene precursors caused an increase in root flavonoid and rhizosphere siderophore while restoring bacterial and fungal microbial cells in roots under combined stress in soil. However, in healthy plants, flavonoid and siderophore levels decreased, accompanied by a reduction in microbial cells to control levels. This suggests that elevated ethylene may regulate root exudates to recruit microbes dominated by host response, aiding soybean plants cope with combined stresses, although this effect may not occur in non-stressed plants. This is the first report on the transcriptional response and physiological adjustments in soybean exposed to alkalinity and drought, potentially advancing knowledge for genetic and agronomic interventions to improve stress resilience in legume crops.

plant biology↗

bioRxiv2024

The effect of constitutive root isoprene emission on root phenotype and physiology under control and salt stress conditions

ABSTRACTIsoprene, a volatile hydrocarbon, is typically emitted from the leaves and other aboveground plant organs; isoprene emission from roots is not well studied. Given its well-known function in plant growth and defense aboveground, isoprene may also be involved in shaping root physiology to resist belowground stress. We used isoprene-emitting transgenic lines (IE) and a non-emitting empty vector and/or wild type lines (NE) of Arabidopsis to elucidate the roles of isoprene in root physiology and salt stress resistance. We assessed root phenotype and metabolic changes, hormone biosynthesis and signaling, and stress-responses under normal and saline conditions of IE and NE lines. We also analyzed the root transcriptome in the presence or absence of salt stress. IE lines emitted isoprene from roots, which was associated with higher primary root growth, root biomass, and root/shoot biomass ratio under both control and salt stress conditions. Transcriptome data indicated that isoprene altered the expression of key genes involved in hormone metabolism and plant responses to stress factors. Our findings reveal that root constitutive isoprene emission sustains root growth also under salinity by regulating and/or priming hormone biosynthesis and signaling mechanisms, amino acids biosynthesis, and expression of key genes relevant to salt stress defense.

plant biology↗

bioRxiv2020

The histidine phosphotransfer AHP4 plays a negative role in Arabidopsis plant response to drought

Cytokinin plays an important role in plant stress responses via a multistep signaling pathway, involving the histidine phosphotransfer proteins (HPs). In Arabidopsis thaliana, the AHP2, AHP3 and AHP5 proteins are known to impact drought responses; however, the role of AHP4 in drought adaptation remains undetermined. In the present study, using a loss-of-function approach we showed that AHP4 possesses a negative regulatory role in Arabidopsiss response to drought. This is evidenced by both higher survival rates of ahp4 than wild-type (WT) plants under drought conditions, and the down-regulated AHP4 expression in WT during periods of dehydration. Comparative transcriptome analysis of ahp4 and WT plants revealed AHP4-mediated expression of several dehydration- and/or abscisic acid (ABA)-responsive genes involved in regulation of various physiological and biochemical processes important for plant drought acclimation. In comparison with WT, ahp4 plants showed increased wax crystal accumulation in stems, thicker cuticles in leaves, greater sensitivity to exogenous ABA at germination, narrow stomatal apertures, heightened leaf temperatures during dehydration, and longer root length under osmotic stress. Additionally, ahp4 plants showed greater photosynthetic efficiency, lower levels of reactive oxygen species (ROS), reduced electrolyte leakage and lipid peroxidation, and increased anthocyanin contents under drought, when compared with WT. These differences displayed in ahp4 plants are likely due to up-regulation of genes that encode enzymes involved in ROS-scavenging and non-enzymatic antioxidant metabolism. The role of AHP4 in negative regulation of multiple protective mechanisms associated with drought tolerance could make editing of AHP4 a promising approach for the production of drought-tolerant crop plants. Significance statementLoss-of-function analysis of the cytokinin signaling member AHP4 revealed its function in Arabidopsis adaptation to drought as a negative regulator, affecting various physiological and biochemical processes by modulating the expression of a large set of genes potentially in a crosstalk with ABA. AHP4 and its homologs are promising candidates for gene editing to develop drought-tolerant crop cultivars.

plant biology↗