Unveiling the molecular mechanisms of arsenic tolerance and resilience in the primitive bryophyte Marchantia polymorpha L.

This study addresses the pressing issue of high arsenic (As) contaminations, which poses a severe threat to various life forms in our ecosystem. Despite this prevailing concern, all organisms have developed some techniques to mitigate the toxic effects of As. Certain plants, such as bryophytes, the earliest land plants, exhibit remarkable tolerance to wide range of harsh environmental conditions, due to their inherent competence. In this study, bryophytes collected from West Bengal, India, across varying contamination levels were investigated for their As tolerance capabilities. Assessment of As accumulation potential and antioxidant defense efficiency, including SOD, CAT, APX, GPX etc. revealed Marchantia polymorpha as the most tolerant species. It exhibited highest As accumulation, antioxidative proficiency, and minimal damage. Transcriptomic analysis of M. polymorpha exposed to 40 µM As(III) for 24 and 48 h identified several early responsive differentially expressing genes (DEGs) associated with As tolerance. These includes GSTs, GRXs, Hsp20s, SULTR1;2, ABCC2 etc., indicating a mechanism involving vacuolar sequestration. Interestingly, one As(III) efflux-transporter ACR3, an extrusion pump, known to combat As toxicity was found to be differentially expressed compared to control. The SEM-EDX analysis, further elucidated the operation of As extrusion mechanism, which contributes added As resilience in M. polymorpha. Yeast complementation assay using Δacr3 yeast cells, showed increased tolerance towards As(III), compared to the mutant cells, indicating As tolerant phenotype. Overall, these findings significantly enhance our understanding of As tolerance mechanisms in bryophytes. This can pave the way for the development of genetically engineered plants with heightened As tolerance and the creation of improved plant varieties.

Functional characterization of rice metallothionein OsMT-I-Id: Insights into metal binding and heavy metal tolerance mechanisms.

Metallothioneins (MTs) are cysteine-rich proteins known for their strong metal-binding capabilities, making them effective in detoxifying heavy metals (HMs). This study focuses on characterizing the functional properties of OsMT-I-Id, a type-I Metallothionein found in rice. Using a HM-responsive yeast cup1Δ (DTY4), ycf1∆ (for cadmium), and acr3∆ mutants (for trivalent arsenic), we assessed the impact of OsMT-I-Id on metal accumulation and cellular resilience. Our results demonstrated that yeast cells expressing OsMT-I-Id showed increased tolerance and accumulated higher levels of copper (Cu), arsenic (As), and cadmium (Cd), compared to control cells. This can be attributed to the protein's ability to chelate and bind HMs. Site-directed mutagenesis was employed to investigate the specific contributions of cysteine residues. The study revealed that yeast cells with a mutated C-domain displayed heightened HM sensitivity, while cells with a mutated N-domain exhibited reduced sensitivity. This underscores the critical role of C-cysteine-rich domains in metal binding and tolerance of type-I rice MTs. Furthermore, the study identified the significance of the 12th cysteine position at the N-domain and the 68th and 72nd cysteine positions at the C-domain in influencing OsMT-I-Id metal-binding capacity. This research provides novel insights into the structure-function relationship and metal binding properties of type-I plant MTs.

A tau class glutathione-S-transferase (OsGSTU5) confers tolerance against arsenic toxicity in rice by accumulating more arsenic in root.

Arsenic (As) considered as one of the hazardous metalloid that hampers various physiological activities in rice. To study the mechanism of As tolerance in rice, one differentially expressed tau class glutathione-S-transferase (OsGSTU5) has been selected and transgenic rice plants with knockdown (KD) and overexpressing (OE) OsGSTU5 were generated. Our results suggested that KD lines became less tolerant to As stress than WT plants, while OE lines showed enhanced tolerance to As. Under As toxicity, OE and KD lines showed enhanced and reduced antioxidant activities such as, SOD, PRX and catalase, respectively indicating its role in ROS homeostasis. In addition, higher malondialdehyde content, poor photosynthetic parameters and higher reactive oxygen species (ROS) in KD plant, suggests that knockdown of OsGSTU5 renders KD plants more susceptible to oxidative damage. Also, the relative expression profile of various transporters such as OsABCC1 (As sequestration), Lsi2 and Lsi6 (As translocaters) and GSH dependent activity of GSTU5 suggests that GSTU5 might help in chelation of As with GSH and sequester it into the root vacuole using OsABCC1 transporter and thus limits the upward translocation of As towards shoot. This study suggests the importance of GSTU5 as a good target to improve the As tolerance in rice.

miRNAs play critical roles in response to abiotic stress by modulating cross-talk of phytohormone signaling.

One of the most interesting signaling molecules that regulates a wide array of adaptive stress responses in plants are the micro RNAs (miRNAs) that are a unique class of non-coding RNAs constituting novel mechanisms of post-transcriptional gene regulation. Recent studies revealed the role of miRNAs in several biotic and abiotic stresses by regulating various phytohormone signaling pathways as well as by targeting a number of transcription factors (TFs) and defense related genes. Phytohormones are signal molecules modulating the plant growth and developmental processes by regulating gene expression. Studies concerning miRNAs in abiotic stress response also show their vital roles in abiotic stress signaling. Current research indicates that miRNAs may act as possible candidates to create abiotic stress tolerant crop plants by genetic engineering. Yet, the detailed mechanism governing the dynamic expression networks of miRNAs in response to stress tolerance remains unclear. In this review, we provide recent updates on miRNA-mediated regulation of phytohormones combating various stress and its role in adaptive stress response in crop plants.