Light-dependent expression and accumulation of miR408-encoded peptide, miPEP408, is regulated by HY5 in Arabidopsis.

Recent studies propose that primary transcripts of miRNAs (pri-miRNAs) contain small Open Reading Frames (ORFs) capable of encoding miRNA-encoded peptides (miPEPs). These miPEPs can function as transcriptional regulators for their corresponding pri-miRNAs, ultimately enhancing mature miRNA accumulation. Notably, pri-miR408 encodes the functional peptide miPEP408, regulating expression of miR408 and its target genes, providing plant tolerance to stresses. While miPEPs are crucial regulators, the factors governing them are have not been studied in detail. Here, we explored the light-dependent regulation of miPEP408 in Arabidopsis. Expression analysis during dark-light transitions revealed light-induced transcription and accumulation of the miPEP408. As the promoter of miR408 contains cis-acting elements responsible for binding to the bZIP-type transcription factor ELONGATED HYPOCOTYL5 (HY5), known for light-mediated regulation in plants, we studied its involvement in the regulation of miR408. Analysis of HY5 mutant (hy5-215), complemented line (HY5OX/hy5), and CONSTITUTIVE PHOTOMORPHOGENIC 1 mutant (cop1-4) plants supported HY5's positive regulation of miPEP408. Grafting and GUS assays further suggested the role of HY5 as a shoot-root mobile signal inducing light-dependent miPEP408 expression. This study underscores the regulatory impact of light on small peptides, exemplified by miPEP408, mediated by the key transcription factor HY5.

microRNA408 and its encoded peptide regulate sulfur assimilation and arsenic stress response in Arabidopsis.

MicroRNAs (miRNAs) are small non-coding RNAs that play a central role in regulating various developmental and biological processes. The expression of miRNAs is differentially modulated in response to various biotic and abiotic stresses. Recent findings have shown that some pri-miRNAs encode small regulatory peptides known as microRNA-encoded peptides (miPEPs). miPEPs regulate the growth and development of plants by modulating corresponding miRNA expression; however, the role of these peptides under different stress conditions remains unexplored. Here, we report that pri-miR408 encodes a small peptide, miPEP408, that regulates the expression of miR408, its targets, and associated phenotype in Arabidopsis. We also report that miR408, apart from Plantacyanin (ARPN) and Laccase3 (LAC3), targets a glutathione S-transferase (GSTU25) that plays a role in sulfur assimilation and exhibits a range of detoxification activities with the environmental pollutant. Plants overexpressing miR408 showed severe sensitivity under low sulfur (LS), arsenite As(III), and LS + As(III) stress, while miR408 mutants developed using the CRISPR/Cas9 approach showed tolerance. Transgenic lines showed phenotypic alteration and modulation in the expression of genes involved in the sulfur reduction pathway and affect sulfate and glutathione accumulation. Similar to miR408 overexpressing lines, the exogenous application of synthetic miPEP408 and miPEP408OX lines led to sensitivity in plants under LS, As(III), and combined LS + As(III) stress compared to the control. This study suggests the involvement of miR408 and miPEP408 in heavy metal and nutrient deficiency responses through modulation of the sulfur assimilation pathway.

Ocimum sanctum, OscWRKY1, regulates phenylpropanoid pathway genes and promotes resistance to pathogen infection in Arabidopsis.

KEY MESSAGE: OscWRKY1 from Ocimum sanctum positively regulates phenylpropanoid pathway genes and rosmarinic acid content. OscWRKY1 overexpression promotes resistance against bacterial pathogen in Arabidopsis. WRKY transcription factor (TF) family regulates various developmental and physiological functions in plants. PAL genes encode enzymes which are involved in plant defense responses, but the direct regulation of PAL genes and phenylpropanoid pathway through WRKY TF's is not well characterized. In the present study, we have characterized an OscWRKY1 gene from Ocimum sanctum which shows induced expression by methyl jasmonate (MeJA), salicylic acid (SA), and wounding. The recombinant OscWRKY1 protein binds to the DIG-labeled (Digoxigenin) W-box cis-element TTGAC[C/T] and activates the LacZ reporter gene in yeast. Overexpression of OscWRKY1 enhances Arabidopsis resistance towards Pseudomonas syringae pv. tomato Pst DC3000. Upstream activator sequences of PAL and C4H have been identified to contain the conserved W-box cis-element (TTGACC) in both O. sanctum and Arabidopsis. OscWRKY1 was found to interact with W-box cis-element present in the PAL and C4H promoters. Silencing of OscWRKY1 using VIGS resulted in reduced expression of PAL, C4H, COMT, F5H and 4CL transcripts. OscWRKY1 silenced plants exhibit reduced PAL activity, whereas, the overexpression lines of OscWRKY1 in Arabidopsis exhibit increased PAL activity. Furthermore, the metabolite analysis of OscWRKY1 silenced plants showed reduced rosmarinic acid content. These results revealed that OscWRKY1 positively regulates the phenylpropanoid pathway genes leading to the alteration of rosmarinic acid content and enhances the resistance against bacterial pathogen in Arabidopsis.

Effect of nanoparticle on rheological properties of surfactant-based nanofluid for effective carbon utilization: capturing and storage prospects.

Previous studies have shown insufficient dispersion and thermal stability of nanofluids for high-temperature carbon capture and storage applications. Compared to the other NPs, TiO2 nanofluids exhibit superior stability due to their high zeta potential. In previous studies, TiO2 nanofluids have shown superior performance in heat transfer and cooling applications along with importing the stability of other nanofluids like SiO2 in form of nanocomposites. Therefore, in this study, a nanofluid formulation consisting of titania nanofluid in a base solution of ethylene glycol (EG) with different co-stabilizers such as surfactants was synthesized for better dispersion stability, enhanced electrical, and rheological properties especially for the use in high-temperature industrial applications which include carbon capture and storage along with enhanced oil recovery. The formulated nanofluid was investigated for stability using dynamic light scattering (DLS) study and electrical conductivity. Additionally, the formulated nanofluid was also examined for thermal stability at high temperatures using an electrical conductivity study followed by rheological measurements at 30 and 90 °C. At a high temperature, the shear-thinning behavior of EG was found highly affected by shear rate; however, this deformation was controlled using TiO2 nanoparticles (NPs). Furthermore, the role of surfactant was also investigated on dispersion stability, electrical conductivity followed by viscosity results, and it was found that the nanofluid is superior in presence of anionic surfactant sodium dodecyl sulfate (SDS) as compared to nonionic surfactant Triton X-100 (TX-100). The inclusion of ionic surfactant provides a charged layer of micelles surrounding the core of a NP and it produced additional surface potential. Consequently, it increases the repulsive force between two adjacent NPs and renders a greater stability to nanofluid while nonionic surfactant allowed monomers to adsorb on the surface of NP via hydrophobic interaction and enhances the short-range interparticle repulsion, to stabilize nanofluid. This makes titania nanofluid suitable for widespread high-temperature applications where conventional nanofluids face limitations. Finally, the application of the synthesized titania nanofluids was explored for the capture and transport of CO2 where the inclusion of the anionic surfactant was found to increase the CO2 capturing ability of titania nanofluids by 140-220% (over the conventional nanofluid) while also showing superior retention at both investigated temperatures. Thus, the study promotes the role of novel surfactant-treated titania nanofluids for carbon removal and storage and recommends their applications involving carbonated fluid injection (CFI) to carbon utilization in oilfield applications.

Light-regulated expression of terpene synthase gene, AtTPS03, is controlled by the bZIP transcription factor, HY5, in Arabidopsis thaliana.

The terpenoid pathway serves as an essential source of all isoprenoid precursors and metabolites that are of great pharmacological importance. The major enzymes for the synthesis of these diverse molecules is the terpene synthases (TPSs), which catalyse the final step of the synthesis of the important secondary products, the terpenes. Previous studies have reported that the various environmental factors, including light govern the synthesis of terpenoids. However, the molecular components and steps involved in the regulation of synthesis of these molecules have not been studied in detail. In this study, we report that the light regulates the expression of the members of terpene synthase gene family in Arabidopsis thaliana. We demonstrate that ELONGATED HYPOCOTYL (HY5), a basic leucine zipper transcription factor, plays a crucial role in light-mediated transcriptional regulation of terpene synthase, AtTPS03. Expression analysis using hy5-215 mutant and HY5 over-expression lines revealed that HY5 acts as a positive regulator of AtTPS03. Additionally, studies including AtTPS03 Promoter::reporter transgenic lines in wild-type and hy5-215, as well as electrophoretic mobility shift assay (EMSA), suggest an interaction of HY5 with the AtTPS03 promoter. Together, our analysis indicate the requirement for HY5 for light-mediated regulation of AtTPS03 for the terpenoid biosynthesis in Arabidopsis.

3'O-Methyltransferase, Ps3'OMT, from opium poppy: involvement in papaverine biosynthesis.

KEY MESSAGE: Using, in silico, in vitro and in planta functional assays, we demonstrate that Ps3'OMT, an 3'-O methyl transferase is linked to papaverine biosynthesis in opium poppy. Papaverine, one of the benzylisoquinoline alkaloids (BIA) synthesized in the medicinally important plant, Papaver somniferum, is known for the potent pharmacological properties. Papaverine biosynthesis has remained debatable as two different pathways, NH (involving N-desmethylated intermediates) and the NCH3 (involving N-methylated intermediates), have been proposed. In addition, there are several intermediate steps in both the proposed pathways that are not very well characterized in terms of specific enzymes. In this study, we report the identification and functional characterization of 3'O-methyltransferase (Ps3'OMT) which might participate in the 3'O-methylation of the intermediates in the papaverine biosynthesis. Comparison of transcript and metabolite profiles of high and low papaverine producing cultivar revealed the occurrence of a 3'O-methyltransferase, Ps3'OMT, which was abundant in aerial organs and shared 72% identity with the GfLOMT7 predicted to have 3'OMT activity. In silico studies based on homology modeling, docking and MD simulations predicted (S)-norlaudanine as the potential substrate forming a stable complex with Ps3'OMT. Suppression of Ps3'OMT through virus-induced gene silencing resulted in a remarkable decrease in the level of papaverine in comparison to control plants. The characterization of the functionally unique Ps3'OMT involved in BIA metabolism suggests an involvement of the NH pathway leading to papaverine biosynthesis.