Does environmental policy stringency improve nature's health in BRICS economies? Implications for sustainable development.

In our groundbreaking exploration, we meticulously delve into the relationship between environmental policy stringency, international trade dynamics, and financial openness within the BRICS group (Brazil, Russia, India, China, and South Africa) spanning from 1996 to 2021. With a focus on critical variables such as economic growth and technological innovation, our empirical findings challenge conventional wisdom. Surprisingly, we found that those stringent environmental policies, when standing alone, do not invariably lead to reduce CO2 emissions. Equally interesting is our startling discovery that the anticipated moderating influence of environmental policy stringency, catalyzed by trade and foreign direct investment, on the well-being of our environment does not materialize; contrarily, both trade and foreign direct investment moderating channels exhibit unanticipated positive correlations with CO2 emissions. These revelations provoke us with the presence of a "pollution haven" phenomenon within the BRICS economies. Furthermore, our investigation reveals that, when examined individually, trade and foreign direct investment also appear to contribute to elevated emission levels. These findings provide a resolute solution to our research quandary, underlining the indispensable requirement for cutting-edge and robust environmental policies. These policies must possess the prowess to effectively counteract the adverse environmental consequences stemming from the amalgamation of global trade and financial integration. In doing so, they shall propel BRICS nations toward a future firmly grounded in principles of sustainability and ecological integrity.

CRISPR/Cas9-mediated mutagenesis of the mediator complex subunits MED5a and MED5b genes impaired secondary metabolite accumulation in hop (Humulus lupulus).

Hop (Humulus lupulus L.) is an important commercial crop known for the biosynthesis of valuable specialized secondary metabolites in glandular trichomes (lupulin glands), which are used for the brewing industry. To achieve burgeoning market demands is the essentiality of comprehensive understanding of the mechanisms of biosynthesis of secondary metabolites in hop. Over the past year, several studies using structural biology and functional genomics approaches have shown that Mediator (MED) serves as an integrative hub for RNAP II-mediated transcriptional regulation of various physiological and cellular processes, including involvement of MED5a and MED5b in hyperaccumulation of phenylpropanoid in A. thaliana. In the present work, an unprecedented attempt was made to generate Hlmed5a/med5b double loci mutant lines in hop using a CRISPR/Cas9-based genome editing system. The Hlmed5a/med5b double loci mutant lines showed reduced expression of structural genes of the flavonoid, humulone, and terpenoid biosynthetic pathways, which was more pronounced in the lupulin gland compared to leaf tissue and was consistent with their reduced accumulation. Phenotypic and anatomical observations revealed that Hlmed5a/med5b double loci mutant line exhibited robust growth, earlier flowering, earlier cone maturity, reduced cone size, variations in floral structure patterns, and distorted lupulin glands without any remarkable changes in leaf morphology, intensity of leaf color, and chlorophyll content. Comparative transcriptome analysis of leaf and lupulin gland tissues indicates that the expression of enzymatic genes related to secondary metabolite biosynthesis, phytohormone biosynthesis, floral organs, flowering time, and trichome development, including other genes related to starch and sucrose metabolism and defense mechanisms, were differentially modulated in the Hlmed5a/med5b lines. The combined results from functional and transcriptomic analyses illuminates the pivotal function of HlMED5a and HlMED5b in homeostasis of secondary meatbolites accumulation in hop.

Benzothiazole-based novel fluorescence probe sensing 1, 3-diaminopropane.

Amines are extensively present in biological systems and are abundantly used in research, industries and agriculture. Systematic detection and quantification of certain amines can help us in food quality control and diagnosis of many diseases. A Schiff base probe HL was designed and successfully synthesized. It was proposed as a sensor for the exclusive detection of 1, 3- diaminopropane through turn-on fluorescence response in a variety of solvents including water. Micromolar limits of detection was achieved in all these solvents. Mechanism of detection was proposed by investigating mass spectrometric and NMR results. These were corroborated with DFT/TD-DFT calculations. Spiking experiments performed in various real water samples revealed the potential of the sensor to be used in day-to-day applications. Paper strip experiments demonstrated the suitability of the probe for real-life applications.

Real time sensor for Fe3+, Al3+, Cu2+ & PPi through quadruple mechanistic pathways using a novel dipodal quinoline-based molecular probe.

A quinoline-based small molecular probe, H2L was designed, synthesized and characterized by different spectroscopic methods. It was utilized as a multi-responsive probe for the detection of Fe3+, Al3+, Cu2+ and PPi. It showed very selective instant turn-on fluorimetric response towards Fe3+and Al3+ with a detection limit in nanomolar range. Solutions of H2L containing Fe3+ or Al3+ could sequentially sense PPi by a turn-off mechanism. Also, H2L could determine the presence of Cu2+ very selectively among a series of other metal ions by a sharp change in colour. Detection of Cu2+ through colorimetry was further investigated by systematic UV-Vis studies and the potential of H2L to act as a potential colorimetric sensor for Cu2+ was suitably established. Filter-paper strip experiments were conducted to demonstrate the practical utility of the proposed sensor. Potential applications of H2L as a sensor for pH in the acidic range has also been explored.

Integrative small RNA and transcriptome analysis provides insight into key role of miR408 towards drought tolerance response in cowpea.

KEY MESSAGE: Drought stress response studies and overexpression of vun-miR408 proved it to be essential for abiotic stress tolerance in cowpea. Small RNA and transcriptome sequencing of an elite high-yielding drought-tolerant Indian cowpea cultivar, Pusa Komal revealed a differential expression of 198 highly conserved, 21 legume-specific, 14 less-conserved, and 10 novel drought-responsive microRNAs (miRNAs) along with 3391 (up-regulated) and 3799 (down-regulated) genes, respectively, in the leaf and root libraries. Among the differentially expressed miRNAs, vun-miR408-3p, showed an up-regulation of 3.53-log2-fold change under drought stress. Furthermore, laccase 12 (LAC 12) was identified as the potential target of vun-miR408-3p using 5' RNA ligase-mediated rapid amplification of cDNA ends. The stable transgenic cowpea lines overexpressing artificial vun-miR408-3p (OX-amiR408) displayed enhanced drought and salinity tolerance as compared to the wild-type plants. An average increase of 30.17% in chlorophyll, 26.57% in proline, and 27.62% in relative water content along with lesser cellular H2O2 level was observed in the transgenic lines in comparison with the wild-type plants under drought stress. Additionally, the scanning electron microscopic study revealed a decrease in the stomatal aperture and an increase in the trichome density in the transgenic lines. The expression levels of laccase 3 and laccase 12, the potential targets of miR408, related to lipid catabolic processes showed a significant reduction in the wild-type plants under drought stress and the transgenic lines, indicating the regulation of lignin content as a plausibly essential trait related to the drought tolerance in cowpea. Taken together, this study primarily focused on identification of drought-responsive miRNAs and genes in cowpea, and functional validation of role of miR408 towards drought stress response in cowpea.

Insight into the cellular and physiological regulatory modulations of Class-I TCP9 to enhance drought and salinity stress tolerance in cowpea.

The Teosinte branched 1/Cycloidea/Proliferating cell factor (TCP) transcription factors are potent growth and developmental regulators in plants, also responsive to various hormonal and environmental stimuli. In this study, we primarily focused on the functional role of TCP9, a nuclear-localised Class-I TCP transcription factor in a drought and heat-tolerant legume crop, cowpea (Vigna unguiculata). Under drought stress, a higher protein expression level of TCP9 was observed in the leaves of the drought-tolerant cowpea cultivar Pusa Komal as compared to the drought-sensitive cultivar TVu-7778. Further, overexpression of VuTCP9 resulted in reduced cell and stomata size, aperture length and width while cell and overall stomatal density in the 35S::VuTCP9 transgenic cowpea lines increased. Phenotypic alterations, such as reduced leaf size and vigour, altered seed coats displaying extension pattern similar to the 'Watson pattern' and delayed senescence were prominent in the transgenic lines. Under normal conditions, the gas exchange and fluorescence measurements indicated reduction in transpiration rate (E), stomatal conductance (gs ) and photosynthetic efficiency (Φ PSII). However, water usage efficiency (WUE) remained unaltered in the transgenic lines as compared to the wild-type (WT) plants. Furthermore, the transgenic lines displayed higher tolerance to oxidative, drought and salinity stress, maintained relatively higher relative water content and lower occurrence of H2 O2 , as compared to the WT plants. Genes related to the jasmonic acid biosynthesis, stomatal development and abiotic stress responsiveness, such as TTG1, NAC25, SPCH and GRP1, increased and LOX2 decreased significantly in the transgenic lines.

Recent progress in the development of MOF-based optical sensors for Fe3.

Fe(iii) is a common pollutant released into our ecosystem from various industrial and anthropogenic activities which when in excess interferes with human health. A plethora of sensors based on various designs and working principles are being continuously synthesized and improvised for its facile detection. In the present review, we have provided a brief overview of the developments made in the field of metal organic framework (MOF) based optical sensors for Fe3+. MOFs have exponentially emerged in the field of research due to their high porosity, modular construction and easy tunability. These inorganic-organic hybrid porous materials are being essentially promoted as optical sensors because of their unique photophysical properties and potential sensing applications.

Study of expressions of miRNAs in the spikelets based on their spatial location on panicle in rice cultivars provided insight into their influence on grain development.

Development of rice cultivars bearing numerous spikelets by breeding approach to increase the yearly production of rice to approximately 800 million metric tons to feed the ever increasing population of the world accompanies poor grain filling in the inferior spikelets preventing achievement of the yield potential. As the initial stages of caryopses development are of much importance for grain filling, spatio-temporal expressions of the miRNAs were studied during these periods in the spikelets of a compact-panicle rice cultivar, Oryza sativa cv. Mahalaxmi, bearing numerous spikelets per panicle to understand the reason of poor grain filling at the level of the initial biochemical events. Differential expression of several known miRNAs between the superior and inferior spikelets suggested great difference in metabolism related to grain filling in the spikelets based on their spatial location on compact panicle. Expressions of five known and four novel miRNAs were validated by Northern. Their targets included the enzymes directly involved in starch biosynthesis like sucrose synthase, starch synthase and pullulanase, besides others. Spatio-temporal expression studies of these miRNAs in the spikelets of Mahalaxmi revealed a pattern of mostly a greater expression in the inferior spikelets compared with the superior ones concomitant with an inverse expression of the target genes, which was not observed in the lax-panicle cultivar Upahar. The study thus revealed that the grain filling in rice is greatly regulated by miRNAs, and these miRNAs or their target genes could be considered for biotechnological interventions for improving grain filling in the rice cultivars of interest.

TICT fluorescent probe for Al3+: Sequential detection of PPi, ATP and ADP in semi-aqueous medium and real-life applications.

A ditopic Schiff base ligand, H2L has been synthesized and characterized by all spectroscopic techniques. It is highly selective and specific towards Al3+ in semi aqueous medium (DMF/H2O mixture) by exhibiting a drastic increase in the fluorescence intensity. The emission studies, spectroscopic data, life time and quantum yield results have been used to understand its binding mode, explore its specificity and establish its efficacy. The intensity difference is remarkable in physiological pH range. Due to its reversible behavior this ditopic fluorescent chemosensor can be used multiple times to make it cost effective. Detection limit for this chemosensor was found to be 0.65 µM. Experiments with TLC plates show that it can be used as a practical and portable sensor for studying environmental samples in real life. The L-Al3+ complex generated in the solution acts as a sensor to sequentially detect pyrophosphate groups present in inorganic pyrophosphates, ATP and ADP among other anions by turning off the fluorescence. Inhibit logic gate and its corresponding truth table has been developed to aid in further exploiting its multidimensional applications.

Identification and characterization of drought responsive miRNAs in a drought tolerant upland rice cultivar KMJ 1-12-3.

Shortfall of rain that creates drought like situation in non-irrigated agriculture system often limits rice production, necessitating introduction of drought tolerance trait into the cultivar of interest. The mechanism governing drought tolerance is, however, largely unknown, particularly the involvement of miRNAs, the master regulators of biochemical events. In this regard, response study on a drought tolerant rice variety KMJ 1-12-3 to 20% PEG (osmolality- 315 mOsm/kg) as drought stress revealed significant changes in abundance of several conserved miRNAs targeting transcription factors like homeodomain-leucine zipper, MADS box family protein, C2H2 zinc finger protein and Myb, well known for their importance in drought tolerance in plants. The response study also revealed significant PEG-induced decrease in abundance of the miRNAs targeting cyclin A, cyclin-dependent kinase, guanine nucleotide exchange factor, GTPase-activating protein, 1-aminocyclopropane-1-carboxylic acid oxidase and indole-3-acetic beta-glucosyl transferase indicating miRNA-regulated role of the cell cycle regulators, G-protein signalling and the plant hormones ethylene and IAA in drought tolerance in plants. The study confirmed the existence of four novel miRNAs, including osa-miR12470, osa-miR12471, osa-miR12472 and osa-miR12473, and the targets of three of them could be successfully validated. The PEG-induced decrease in abundance of the novel miRNAs osa-miR12470 and osa-miR12473 targeting RNA dependent RNA polymerase and equilibrative nucleoside transporter, respectively suggested an overall increase in both degradation and synthesis of nucleic acids in plants challenged with drought stress. The drought-responsive miRNAs identified in the study may be proved useful in introducing the trait in the rice cultivars of choice by manipulation of their cellular abundance.

Cowpea [Vigna unguiculata (L.) Walp].

Agrobacterium tumefaciens-mediated transformation is an efficient method for incorporating genes and recovering stable transgenic plants in cowpea because this method offers several advantages such as the defined integration of transgenes, potentially low copy number, and preferential integration into transcriptional active regions of the chromosome. Cotyledonary node explants of cowpea present an attractive target for T-DNA delivery followed by regeneration of shoots via axillary proliferation without involvement of a de novo regeneration pathway. In this chapter, we describe a detailed protocol for Agrobacterium-mediated transformation of the cowpea variety Pusa Komal. The seedling cotyledonary node explants are used for cocultivation with an Agrobacterium strain EHA105 harboring standard binary vector, pCAMBIA2301 or pNOV2819, and putative transformed plants are selected using aminoglycoside antibiotic or mannose as sole carbon source, respectively. The entire process includes explant infection to transgenic seed generation in greenhouse.

Cloning and functional characterization of a vacuolar Na+/H+ antiporter gene from mungbean (VrNHX1) and its ectopic expression enhanced salt tolerance in Arabidopsis thaliana.

Plant vacuolar NHX exchangers play a significant role in adaption to salt stress by compartmentalizing excess cytosolic Na+ into vacuoles and maintaining cellular homeostasis and ionic equilibrium. We cloned an orthologue of the vacuolar Na+/H+ antiporter gene, VrNHX1 from mungbean (Vigna radiata), an important Asiatic grain legume. The VrNHX1 (Genbank Accession number JN656211.1) contains 2095 nucleotides with an open reading frame of 1629 nucleotides encoding a predicted protein of 542 amino acids with a deduced molecular mass of 59.6 kDa. The consensus amiloride binding motif (84LFFIYLLPPI93) was observed in the third putative transmembrane domain of VrNHX1. Bioinformatic and phylogenetic analysis clearly suggested that VrNHX1 had high similarity to those of orthologs belonging to Class-I clade of plant NHX exchangers in leguminous crops. VrNHX1 could be strongly induced by salt stress in mungbean as the expression in roots significantly increased in presence of 200 mM NaCl with concomitant accumulation of total [Na+]. Induction of VrNHX1 was also observed under cold and dehydration stress, indicating a possible cross talk between various abiotic stresses. Heterologous expression in salt sensitive yeast mutant AXT3 complemented for the loss of yeast vacuolar NHX1 under NaCl, KCl and LiCl stress indicating that VrNHX1 was the orthologue of ScNHX1. Further, AXT3 cells expressing VrNHX1 survived under low pH environment and displayed vacuolar alkalinization analyzed using pH sensitive fluorescent dye BCECF-AM. The constitutive and stress inducible expression of VrNHX1 resulted in enhanced salt tolerance in transgenic Arabidopsis thaliana lines. Our work suggested that VrNHX1 was a salt tolerance determinant in mungbean.

Arsenic stress in rice: redox consequences and regulation by iron.

Arsenic (As) contamination is a serious hazard to human health and agriculture. It has emerged as an important threat for rice cultivation mainly in South Asian countries. In this study, we investigated the effect of iron (Fe) supplementation on arsenic (As(V)) induced oxidative stress responses in rice (Oryza sativa L.). Rice seedlings treated with As(V) for 24 and 48 h in presence or absence of 2.5 mM Fe after which the root and shoot tissues were harvested for analysis. The results indicate significant (p ≤ 0.05) reduction in root and shoot length/dry biomass. Supplementation of Fe showed improved growth responses under stress as compared to As(V) alone. The scanning electron microscopy (SEM) analysis of roots under As(V) treatment for 48 h showed major alterations in root structure and integrity, although no noticeable changes were observed in Fe - supplemented seedlings. Significantly high (p ≤ 0.05) accumulation of As(V) was observed in root and shoot after 24 and 48 h of stress. However, under Fe - supplementation As accumulation in root and shoot were considerably low after 24 and 48 h of As(V) treatment. The hydrogen peroxide (H2O2) and malondialdehyde (MDA) content in both root and shoot increased significantly (p ≤ 0.05) after 24 and 48 h of As(V) treatment. In Fe - supplemented seedlings, the levels of H2O2 and MDA were considerably low as compared to As(V) alone. Ascorbate (AsA) and glutathione (GSH) levels also increased significantly (p ≤ 0.05) under As(V) stress as compared to control and Fe-supplemented seedlings. Activities of catalase (CAT) and superoxide dismutase (SOD) were significantly (p ≤ 0.05) high after 24 and 48 h of As(V) treatment as compared to Fe-supplemented seedlings. The gene expression analysis revealed up-regulation of metallothionein (MT1, MT2) and nodulin 26-like intrinsic protein (NIP2;1) genes after 5d of As treatment, while their expressions were repressed under Fe-supplementation. Our results indicate that Fe regulates oxidative stress and promotes growth under As stress.

Successful recovery of transgenic cowpea (Vigna unguiculata) using the 6-phosphomannose isomerase gene as the selectable marker.

A new method for obtaining transgenic cowpea was developed using positive selection based on the Escherichia coli 6-phosphomannose isomerase gene as the selectable marker and mannose as the selective agent. Only transformed cells were capable of utilizing mannose as a carbon source. Cotyledonary node explants from 4-day-old in vitro-germinated seedlings of cultivar Pusa Komal were inoculated with Agrobacterium tumefaciens strain EHA105 carrying the vector pNOV2819. Regenerating transformed shoots were selected on medium supplemented with a combination of 20 g/l mannose and 5 g/l sucrose as carbon source. The transformed shoots were rooted on medium devoid of mannose. Transformation efficiency based on PCR analysis of individual putative transformed shoots was 3.6%. Southern blot analysis on five randomly chosen PCR-positive plants confirmed the integration of the pmi transgene. Qualitative reverse transcription (qRT-PCR) analysis demonstrated the expression of pmi in T0 transgenic plants. Chlorophenol red (CPR) assays confirmed the activity of PMI in transgenic plants, and the gene was transmitted to progeny in a Mendelian fashion. The transformation method presented here for cowpea using mannose selection is efficient and reproducible, and could be used to introduce a desirable gene(s) into cowpea for biotic and abiotic stress tolerance.

Improved Agrobacterium-mediated transformation of cowpea via sonication and vacuum infiltration.

An improved method of Agrobacterium-mediated transformation of cowpea was developed employing both sonication and vacuum infiltration treatments. 4 day-old cotyledonary nodes were used as explants for co-cultivation with Agrobacterium tumefaciens strain EHA105 harbouring the binary vector pSouv-cry1Ac. Among the different injury treatments, vacuum infiltration and their combination treatments tested, sonication for 20 s followed by vacuum infiltration for 5 min with A. tumefaciens resulted in highest transient GUS expression efficiency (93% explants expressing GUS at regenerating sites). After 3 days of co-cultivation, the explants were cultured in 150 mg/l kanamycin-containing selection medium and putative transformed plants were recovered. The presence, integration and expression of nptII and cry1Ac genes in T0 transgenic plants were confirmed by polymerase chain reaction (PCR), genomic Southern and qualitative reverse transcription (RT)-PCR analysis. Western blot hybridization and enzyme-linked immunosorbent assay (ELISA) detected and demonstrated the accumulation of Cry1Ac protein in transgenic plants. The cry1Ac gene transmitted in a Mendelian fashion. The stable transformation efficiency increased by 88.4% using both sonication-assisted Agrobacterium-mediated transformation (SAAT) and vacuum infiltration than simple Agrobacterium-mediated transformation in cowpea.

Histology of vitamin A induced ectopic limbs and normal hind limbs in tadpoles of Polypedates maculatus.

Marked histological similarities were observed between normal and vitamin A induced ectopic limb buds of P. maculatus. However, close association of nephric tubule and lateral plate mesoderm, as seen in normal hind limb bud does not seem to be essential for ectopic limb development. The ectopic limbs tend to develop in pairs.