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1.
Development ; 151(12)2024 Jun 15.
Article in English | MEDLINE | ID: mdl-38814747

ABSTRACT

The shoot apical meristem (SAM) gives rise to the aboveground organs of plants. The size of the SAM is relatively constant due to the balance between stem cell replenishment and cell recruitment into new organs. In angiosperms, the transcription factor WUSCHEL (WUS) promotes stem cell proliferation in the central zone of the SAM. WUS forms a negative feedback loop with a signaling pathway activated by CLAVATA3 (CLV3). In the periphery of the SAM, the ERECTA family receptors (ERfs) constrain WUS and CLV3 expression. Here, we show that four ligands of ERfs redundantly inhibit the expression of these two genes. Transcriptome analysis confirmed that WUS and CLV3 are the main targets of ERf signaling and uncovered new ones. Analysis of promoter reporters indicated that the WUS expression domain mostly overlaps with the CLV3 domain and does not shift along the apical-basal axis in clv3 mutants. Our three-dimensional mathematical model captured gene expression distributions at the single-cell level under various perturbed conditions. Based on our findings, CLV3 regulates cellular levels of WUS mostly through autocrine signaling, and ERfs regulate the spatial expression of WUS, preventing its encroachment into the peripheral zone.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Gene Expression Regulation, Plant , Homeodomain Proteins , Meristem , Signal Transduction , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis/growth & development , Meristem/metabolism , Meristem/genetics , Arabidopsis Proteins/metabolism , Arabidopsis Proteins/genetics , Signal Transduction/genetics , Homeodomain Proteins/metabolism , Homeodomain Proteins/genetics , Receptors, Cell Surface/metabolism , Receptors, Cell Surface/genetics , Transcription Factors/metabolism , Transcription Factors/genetics , Protein Serine-Threonine Kinases/metabolism , Protein Serine-Threonine Kinases/genetics , Models, Biological
2.
Funct Integr Genomics ; 23(1): 66, 2023 Feb 25.
Article in English | MEDLINE | ID: mdl-36840774

ABSTRACT

Mutations in Photoperiod-1 (Ppd-1) gene are known to modify flowering time and yield in wheat. We cloned TaPpd-1 from wheat and found high similarity among the three homoeologs of TaPpd-1. To clarify the characteristics of TaPpd-1 homoeologs in different photoperiod conditions for inflorescence architecture and yield, we used CRISPR/Cas9 system to generate Tappd-1 mutant plants by simultaneous modification of the three homoeologs of wheat Ppd-1. Tappd-1 mutant plants showed no off-target mutations. Four T0-edited lines under short-day length and three lines under long-day length conditions with the mutation frequency of 25% and 21%, respectively. These putative transgenic plants of all the lines were self-fertilized and generated T1 and T2 progenies and were evaluated by phenotypic and expression analysis. Results demonstrated that simultaneously edited TaPpd-1- A1, B1, and D1 homoeologs gene copies in T2_SDL-8-4, T2_SDL-4-5, T2_SDL-3-9, and T2_LDL-10-9 showed similar spike inflorescence, flowering time, and significantly increase in 1000-grain weight, grain area, grain width, grain length, plant height, and spikelets per spike due to mutation in both alleles of Ppd-B1 and Ppd-D1 homoeologs but only spike length was decreased in T2_SDL-8-4, T2_SDL-4-5, and T2_LDL-13-3 mutant lines due to mutation in both alleles of Ppd-A1 homoeolog under both conditions. Our results indicate that all TaPpd1 gene homoeologs influence wheat spike development by affecting both late flowering and earlier flowering but single mutant TaPpd-A1 homoeolog affect lowest as compared to the combination with double mutants of TaPpd-B1 and TaPpd-D1, TaPpd-A1 and TaPpd-B1, and TaPpd-A1 and TaPpd-D1 homoeologs for yield enhancement. Our findings further raised the idea that the relative expression of the various genomic copies of TaPpd-1 homoeologs may have an impact on the spike inflorescence architecture and grain morphometric features in wheat cultivars.


Subject(s)
Photoperiod , Triticum , Triticum/genetics , CRISPR-Cas Systems , Phenotype , Edible Grain/genetics
3.
Funct Integr Genomics ; 23(2): 119, 2023 Apr 06.
Article in English | MEDLINE | ID: mdl-37022538

ABSTRACT

Genome editing is a useful, adaptable, and favored technique for both functional genomics and crop enhancement. Over the years, rapidly evolving genome editing technologies, including clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas), transcription activator-like effector nucleases (TALENs), and zinc finger nucleases (ZFNs), have shown broad application prospects in gene function research and improvement of critical agronomic traits in many crops. These technologies have also opened up opportunities for plant breeding. These techniques provide excellent chances for the quick modification of crops and the advancement of plant science in the future. The current review describes various genome editing techniques and how they function, particularly CRISPR/Cas9 systems, which can contribute significantly to the most accurate characterization of genomic rearrangement and plant gene functions as well as the enhancement of critical traits in field crops. To accelerate the use of gene-editing technologies for crop enhancement, the speed editing strategy of gene-family members was designed. As it permits genome editing in numerous biological systems, the CRISPR technology provides a valuable edge in this regard that particularly captures the attention of scientists.


Subject(s)
CRISPR-Cas Systems , Genetic Engineering , Plants, Genetically Modified/genetics , Genetic Engineering/methods , Genome, Plant , Plant Breeding/methods , Crops, Agricultural/genetics , Technology
4.
BMC Plant Biol ; 23(1): 250, 2023 May 12.
Article in English | MEDLINE | ID: mdl-37173631

ABSTRACT

BACKGROUND: Fatty acid desaturases (FADs) are involved in regulating plant fatty acid composition by adding double bonds to growing hydrocarbon chain. Apart from regulating fatty acid composition FADs are of great importance, and are involved in stress responsiveness, plant development, and defense mechanisms. FADs have been extensively studied in crop plants, and are broadly classed into soluble and non-soluble fatty acids. However, FADs have not yet been characterized in Brassica carinata and its progenitors. RESULTS: Here we have performed comparative genome-wide identification of FADs and have identified 131 soluble and 28 non-soluble FADs in allotetraploid B. carinata and its diploid parents. Most soluble FAD proteins are predicted to be resided in endomembrane system, whereas FAB proteins were found to be localized in chloroplast. Phylogenetic analysis classed the soluble and non-soluble FAD proteins into seven and four clusters, respectively. Positive type of selection seemed to be dominant in both FADs suggesting the impact of evolution on these gene families. Upstream regions of both FADs were enriched in stress related cis-regulatory elements and among them ABRE type of elements were in abundance. Comparative transcriptomic data analysis output highlighted that FADs expression reduced gradually in mature seed and embryonic tissues. Moreover, under heat stress during seed and embryo development seven genes remained up-regulated regardless of external stress. Three FADs were only induced under elevated temperature whereas five genes were upregulated under Xanthomonas campestris stress suggesting their involvement in abiotic and biotic stress response. CONCLUSIONS: The current study provides insights into the evolution of FADs and their role in B. carinata under stress conditions. Moreover, the functional characterization of stress-related genes would exploit their utilization in future breeding programs of B. carinata and its progenitors.


Subject(s)
Brassica , Transcriptome , Fatty Acid Desaturases/genetics , Fatty Acid Desaturases/metabolism , Brassica/genetics , Brassica/metabolism , Phylogeny , Plant Breeding , Fatty Acids , Gene Expression Regulation, Plant
5.
Mol Pharm ; 20(2): 1382-1393, 2023 02 06.
Article in English | MEDLINE | ID: mdl-36583939

ABSTRACT

The study aimed to improve the treatment of impetigo with naturally occurring quercetin and its copper-quercetin (Cu-Q) complex by preparing sustained-release (SR) nanoparticles of polycaprolactone (PCL). The solvent evaporation method was used for the copper-quercetin (Cu-Q) complex formation, and their PCL nanoparticles (PCL-NPs, Q-PCL-NPs, and Cu-Q-PCL-NPs) were prepared by the high-pressure homogenization method. Synthesis of nanoparticles was confirmed by their physicochemical and antibacterial properties of quercetin against Gram-positive as well as Gram-negative bacteria. The percentage loading efficiency of quercetin and release in 100 mM of phosphate buffer pH 7.4 and 5.5 at 37 °C was found to be more than 90% after 24 h with the zero-order release pattern. Minimum inhibitory concentration of nanoparticles was found to increase threefold in the case of Cu-Q-PCL-NPs may be due to the synergistic antibacterial behavior. Scanning electron microscopy showed spherical nanoparticles, and surface roughness was confirmed by atomic force microscopy analysis. Fortunately, no sign of irritation on rat skin even at 3%, was seen. In vitro antioxidant assay by 2,2-diphenyl-1-picrylhydrazyl reduction was found to be ≤80 ± 0.02% which confirmed their scavenging activity. Interestingly, for the ex vivo study, the tape-stripping model was applied against Staphylococcus aureus containing rats and showed the formation of the epidermal layer within 4-5 days. Confirmation of antibacterial activity of pure quercetin, from Cu-Q complex, and their SR release from Q-PCL-NPs and Cu-Q-PCL-NPs was considered an effective tool for the treatment of skin diseases and can be used as an alternative of already resistant ciprofloxacin in impetigo.


Subject(s)
Impetigo , Nanoparticles , Rats , Animals , Quercetin/pharmacology , Quercetin/therapeutic use , Quercetin/chemistry , Copper/chemistry , Delayed-Action Preparations , Nanoparticles/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry
6.
Mol Biol Rep ; 50(10): 7967-7979, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37535247

ABSTRACT

BACKGROUND: Millions of people around the globe are affected by Alzheimer's disease (AD). This crippling condition has no treatment despite intensive studies. Some phytocompounds have been shown to protect against Alzheimer's in recent studies. METHODS: Thus, this work aimed to examine Bacopa monnieri phytocompounds' synergistic effects on neurodegeneration, antioxidant activity, and cognition in the scopolamine-induced AD mice model. The toxicity study of two phytocompounds: quercetin and bacopaside X revealed an LD50 of more than 2000 mg/kg since no deaths occurred. RESULTS: The neuroprotection experiment consists of 6 groups i.e., control (saline), scopolamine (1 mg/kg), donepezil (5 mg/kg), Q (25 mg/kg), BX (20 mg/kg), and Q + BX (25 mg/kg + 20 mg/kg). Visual behavioral assessment using the Morris water maze showed that animals in the diseased model group (scopolamine) moved more slowly toward the platform and exhibited greater thigmotaxis behavior than the treatment and control groups. Likewise, the concentration of biochemical NO, GSH, and MDA improved in treatment groups concerning the diseased group. mRNA levels of different marker genes including ChAT, IL-1α, IL-1 ß, TNF α, tau, and ß secretase (BACE1) improved in treatment groups with respect to the disease group. CONCLUSION: Both bacopaside X and quercetin synergistically have shown promising results in neuroprotection. Therefore, it is suggested that Q and BX may work synergistically due to their antioxidant and neuroprotective property.


Subject(s)
Alzheimer Disease , Bacopa , Neuroprotective Agents , Humans , Mice , Animals , Alzheimer Disease/chemically induced , Alzheimer Disease/drug therapy , Scopolamine/pharmacology , Scopolamine/therapeutic use , Bacopa/chemistry , Amyloid Precursor Protein Secretases , Quercetin/pharmacology , Quercetin/therapeutic use , Neuroprotection , Neuroprotective Agents/pharmacology , Neuroprotective Agents/therapeutic use , Aspartic Acid Endopeptidases , Antioxidants/pharmacology , Antioxidants/therapeutic use , Maze Learning
7.
Mol Biol Rep ; 50(9): 7381-7392, 2023 Sep.
Article in English | MEDLINE | ID: mdl-37450076

ABSTRACT

BACKGROUND: Alkaline-salt is one of the abiotic stresses that slows plant growth and developmental processes and threatens crop yield. Long non-coding RNAs (lncRNAs) are endogenous RNA found in plants that engage in a variety of cellular functions and stress responses. METHOD: lncRNAs act as competing endogenous RNAs (ceRNA) and constitute a new set of gene control. The precise regulatory mechanism by which lncRNAs function as ceRNAs in response to alkaline-salt stress remains unclear. We identified alkaline-salt responsive lncRNAs using transcriptome-wide analysis of two varieties including alkaline-salt tolerant [WD20342 (WD)] and alkaline-salt sensitive [Caidao (CD)] rice cultivar under control and alkaline-salt stress treated [WD20342 (WDT, and Caidao (CDT)] conditions. RESULTS: Investigating the competitive relationships between mRNAs and lncRNAs, we next built a ceRNA network involving lncRNAs based on the ceRNA hypothesis. Expression profiles revealed that a total of 65, 34, and 1549 differentially expressed (DE) lncRNAs, miRNAs, and mRNAs were identified in alkaline-salt tolerant WD (Control) vs. WDT (Treated). Similarly, 75 DE-lncRNAs, 34 DE-miRNAs, and 1725 DE-mRNAs (including up-regulated and down-regulated) were identified in alkaline-salt sensitive CD (Control) vs. CDT (Treated), respectively. An alkaline-salt stress ceRNA network discovered 321 lncRNA-miRNA-mRNA triplets in CD and CDT, with 32 lncRNAs, 121 miRNAs, and 111 mRNAs. Likewise, 217 lncRNA-miRNA-mRNA triplets in WD and WDT revealed the NONOSAT000455-osa_miR5809b-LOC_Os11g01210 triplet with the highest degree as a hub node with the most significant positive correlation in alkaline-salt stress response. CONCLUSION: The results of our investigation indicate that osa-miR5809b is dysregulated and plays a part in regulating the defense response of rice against alkaline-salt stress. Our study highlights the regulatory functions of lncRNAs acting as ceRNAs in the mechanisms underlying alkaline-salt resistance in rice.


Subject(s)
MicroRNAs , Oryza , RNA, Long Noncoding , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Oryza/genetics , Oryza/metabolism , Gene Regulatory Networks , MicroRNAs/genetics , MicroRNAs/metabolism , Salt Stress/genetics , RNA, Messenger/genetics
8.
Mol Biol Rep ; 51(1): 22, 2023 Dec 18.
Article in English | MEDLINE | ID: mdl-38110786

ABSTRACT

BACKGROUND: Salinity is one of the main abiotic factors that restrict plant growth, physiology, and crop productivity is salt stress. About 33% of the total irrigated land suffers from severe salinity because of intensive underground water extraction and irrigation with brackish water. Thus, it is important to understand the genetic mechanism and identify the novel genes involved in salt tolerance for the development of climate-resilient rice cultivars. METHODS AND RESULTS: In this study, two rice genotypes with varying tolerance to salt stress were used to investigate the differential expressed genes and molecular pathways to adapt under saline soil by comparative RNA sequencing at 42 days of the seedling stage. Salt-susceptible (S3) and -tolerant (S13) genotypes revealed 3982 and 3463 differentially expressed genes in S3 and S13 genotypes. The up-regulated genes in both genotypes were substantially enriched in different metabolic processes and binding activities. Biosynthesis of secondary metabolites, phenylpropanoid biosynthesis, and plant signal transduction mechanisms were highly enriched. Salt-susceptible and -tolerant genotypes shared the same salt adaptability mechanism with no significant quantitative differences at the transcriptome level. Moreover, bHLH, ERF, NAC, WRKY, and MYB transcription factors were substantially up-regulated under salt stress. 391 out of 1806 identified novel genes involved in signal transduction mechanisms. Expression profiling of six novel genes further validated the findings from RNA-seq data. CONCLUSION: These findings suggest that the differentially expressed genes and molecular mechanisms involved in salt stress adaptation are conserved in both salt-susceptible and salt-tolerant rice genotypes. Further molecular characterization of novel genes will help to understand the genetic mechanism underlying salt tolerance in rice.


Subject(s)
Oryza , Transcriptome , Transcriptome/genetics , Oryza/metabolism , Gene Expression Profiling , Salt Stress , Genotype , Gene Expression Regulation, Plant/genetics , Stress, Physiological/genetics
9.
New Phytol ; 234(1): 122-136, 2022 04.
Article in English | MEDLINE | ID: mdl-34716593

ABSTRACT

Shoot branching is regulated by multiple signals. Previous studies have indicated that sucrose may promote shoot branching through suppressing the inhibitory effect of the hormone strigolactone (SL). However, the molecular mechanisms underlying this effect are unknown. Here, we used molecular and genetic tools to identify the molecular targets underlying the antagonistic interaction between sucrose and SL. We showed that sucrose antagonizes the suppressive action of SL on tillering in rice and on the degradation of D53, a major target of SL signalling. Sucrose inhibits the gene expression of D3, the orthologue of the Arabidopsis F-box MAX2 required for SL signalling. Overexpression of D3 antagonizes sucrose inhibition of D53 degradation and enables the SL inhibition of tillering under high sucrose. Sucrose prevents SL-induced degradation of D14, the SL receptor involved in D53 degradation. In contrast to D3, D14 overexpression enhances D53 protein levels and sucrose-induced tillering, even in the presence of SL. Our results show that sucrose inhibits SL response by affecting key components of SL signalling and, together with previous studies reporting the inhibition of SL synthesis by nitrate and phosphate, demonstrate the central role played by SLs in the regulation of plant architecture by nutrients.


Subject(s)
Arabidopsis , Oryza , Arabidopsis/genetics , Gene Expression Regulation, Plant , Lactones/metabolism , Lactones/pharmacology , Oryza/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Sucrose/metabolism , Sucrose/pharmacology
10.
Plant Physiol ; 186(1): 497-518, 2021 05 27.
Article in English | MEDLINE | ID: mdl-33591317

ABSTRACT

Leaf morphology influences photosynthesis, transpiration, and ultimately crop yield. However, the molecular mechanism of leaf development is still not fully understood. Here, we identified and characterized the narrow leaf21 (nal21) mutant in rice (Oryza sativa), showing a significant reduction in leaf width, leaf length and plant height, and increased tiller number. Microscopic observation revealed defects in the vascular system and reduced epidermal cell size and number in the nal21 leaf blade. Map-based cloning revealed that NAL21 encodes a ribosomal small subunit protein RPS3A. Ribosome-targeting antibiotics resistance assay and ribosome profiling showed a significant reduction in the free 40S ribosome subunit in the nal21 mutant. The nal21 mutant showed aberrant auxin responses in which multiple auxin response factors (ARFs) harboring upstream open-reading frames (uORFs) in their 5'-untranslated region were repressed at the translational level. The WUSCHEL-related homeobox 3A (OsWOX3A) gene, a key transcription factor involved in leaf blade lateral outgrowth, is also under the translational regulation by RPS3A. Transformation with modified OsARF11, OsARF16, and OsWOX3A genomic DNA (gDNA) lacking uORFs rescued the narrow leaf phenotype of nal21 to a better extent than transformation with their native gDNA, implying that RPS3A could regulate translation of ARFs and WOX3A through uORFs. Our results demonstrate that proper translational regulation of key factors involved in leaf development is essential to maintain normal leaf morphology.


Subject(s)
Oryza/genetics , Plant Leaves/growth & development , Plant Proteins/genetics , Ribosomal Proteins/genetics , Oryza/growth & development , Plant Leaves/genetics , Plant Proteins/metabolism , Ribosomal Proteins/metabolism
11.
Physiol Plant ; 174(3): e13696, 2022 May.
Article in English | MEDLINE | ID: mdl-35502736

ABSTRACT

Globally, rice is being consumed as a main staple food and faces different kinds of biotic and abiotic stresses such drought, salinity, and pest attacks. Through the cytokinin signaling, Type-B authentic response regulators (ARR-Bs) respond positively towards the environmental stimuli. ARR-Bs are involved in abiotic stress tolerance and plant development but their molecular mechanisms in fragrant rice are still not fully explored. The current study showed the genome-wide characterization of OsARR-B genes under alkaline salt stress. Results showed that in total, 24 OsARR-B genes were found and divided into four subgroups on the basis of a phylogenetic analysis. These genes were located on all rice chromosomes except 8 and 10. Analysis of gene duplications, gene structure, cis-elements, protein-protein interactions, and miRNA were performed. Gene ontology analysis showed that OsARR-B genes are involved in plant development through the regulation of molecular functions, biological processes, and cellular components. Furthermore, 117 and 192 RNA editing sites were detected in chloroplast and mitochondrial genes, respectively, encoding proteins of OsARR-B. In chloroplast and mitochondrial genes, six and nine types of amino acid changes, respectively, were caused by RNA editing, showing that RNA editing has a role in the alkaline salt stress tolerance in fragrant rice. We also used a comparative transcriptome approach to study the gene expression changes in alkaline tolerant and susceptible genotypes. Under alkaline salt stress, OsARR-B5, OsARR-B7, OsARR-B9, OsARR-B10, OsARR-B16, OsARR-B22, and OsARR-B23 showed higher transcript levels in alkaline salt tolerant genotypes as compared to susceptible ones. Quantitative RT-PCR showed upregulation of gene expression in the alkaline tolerant genotypes under alkaline stress. Our study explored the gene expression profiling and RESs of two rice contrasting genotypes, which will help to understand the molecular mechanisms of alkaline salt tolerance in fragrant rice.


Subject(s)
Oryza , Gene Expression Profiling , Gene Expression Regulation, Plant/genetics , Genes, Regulator , Oryza/metabolism , Phylogeny , Plant Proteins/genetics , Plant Proteins/metabolism , Salt Stress/genetics , Salt Tolerance/genetics , Stress, Physiological/genetics
12.
Biol Cybern ; 116(5-6): 661-685, 2022 12.
Article in English | MEDLINE | ID: mdl-36305942

ABSTRACT

Detecting small moving targets against a cluttered background in visual data is a challenging task. The main problems include spatio-temporal target contrast enhancement, background suppression and accurate target segmentation. When targets are at great distances from a non-stationary camera, the difficulty of these challenges increases. In such cases the moving camera can introduce large spatial changes between frames which may cause issues in temporal algorithms; furthermore targets can approach a single pixel, thereby affecting spatial methods. Previous literature has shown that biologically inspired methods, based on the vision systems of insects, are robust to such conditions. It has also been shown that the use of divisive optic-flow inhibition with these methods enhances the detectability of small targets. However, the location within the visual pathway the inhibition should be applied was ambiguous. In this paper, we investigated the tunings of some of the optic-flow filters and use of a nonlinear transform on the optic-flow signal to modify motion responses for the purpose of suppressing false positives and enhancing small target detection. Additionally, we looked at multiple locations within the biologically inspired vision (BIV) algorithm where inhibition could further enhance detection performance, and look at driving the nonlinear transform with a global motion estimate. To get a better understanding of how the BIV algorithm performs, we compared to other state-of-the-art target detection algorithms, and look at how their performance can be enhanced with the optic-flow inhibition. Our explicit use of the nonlinear inhibition allows for the incorporation of a wider dynamic range of inhibiting signals, along with spatio-temporal filter refinement, which further increases target-background discrimination in the presence of camera motion. Extensive experiments shows that our proposed approach achieves an improvement of 25% over linearly conditioned inhibition schemes and 2.33 times the detection performance of the BIV model without inhibition. Moreover, our approach achieves between 10 and 104 times better detection performance compared to any conventional state-of-the-art moving object detection algorithm applied to the same, highly cluttered and moving scenes. Applying the nonlinear inhibition to other algorithms showed that their performance can be increased by up to 22 times. These findings show that the application of optic-flow- based signal suppression should be applied to enhance target detection from moving platforms. Furthermore, they indicate where best to look for evidence of such signals within the insect brain.


Subject(s)
Motion Perception , Optic Flow , Animals , Motion , Motion Perception/physiology , Vision, Ocular , Algorithms , Insecta
13.
Bioorg Chem ; 129: 106196, 2022 12.
Article in English | MEDLINE | ID: mdl-36279741

ABSTRACT

The h-NTPDases is an essential family of ectonucleotidases that consists of eight isozymes with various physiological functions. The undesired activity of the h-NTPDases leads to pathological conditions such as cancer, diabetes, inflammation, and thrombosis. In the present study, a series of thienopyrimidines was synthesized employing a sequential SNAr and Suzuki coupling to synthesize diverse aryl substituted thienopyrimidine glycinate derivatives. The synthesized compounds constituted electron donating, electron-deficient, heteroaryl, and fluorinated substituents. The thienopyrimidines were screened against h-NTPDases to determine the effect on the activity of the h-NTPDases-1, -2, -3, and -8. The compound 3j selectively blocked the isozyme h-NTPDases1, while the compounds 3e, 3m, and 4a were selective inhibitors of h-NTPDases2. The activity of the isozyme h-NTPDases3 was selectively reduced by inhibitor 3k whereas, the compound 3d was found as the most active inhibitor against isozyme h-NTPDase8. The molecular docking study interpreted the interactions of the potent inhibitors of the respective isozymes with important amino acid residues i.e., Asp54, Ser57, His59, Ser58, His59, Asp213, and Phe360 of h-NTPDases1 protein; residues Arg 392, Ala393, Ala347, Tye350 and Arg245 of h-NTPDases2; amino acids Arg67, Ser65, Ala323, Gly222, and Tyr375 of h-NTPDases3 whereas in case of h-NTPDases8, the residues Val436, Gln74, Gly179, and Val71 were involved in interaction with the inhibitors docked into the active sites of these isozymes.


Subject(s)
Isoenzymes , Pyrimidines , Molecular Docking Simulation , Structure-Activity Relationship , Pyrimidines/pharmacology , Molecular Structure
14.
Genomics ; 113(1 Pt 2): 1029-1043, 2021 01.
Article in English | MEDLINE | ID: mdl-33157261

ABSTRACT

Rice is an important cereal crop that serves as staple food for more than half of the world population. Abiotic stresses resulting from changing climatic conditions are continuously threating its yield and production. Genes in APETALA-2 (AP2) family encode transcriptional regulators implicated during regulation of developmental processes and abiotic stress responses but their identification and characterization in indica rice was still missing. In this context, twenty-six genes distributed among eleven chromosomes in Indica rice encoding AP2 transcription-factor subfamily were identified and their diverse haplotypes were studied. Phylogenetic analysis of OsAP2 TF family-members grouped them into three clades indicating conservation of clades among cereals. Segmental duplications were observed to be principal route of evolution, supporting the higher positive selection-pressure, which were estimated to be originated about 10.57 to 56.72 million years ago (MYA). Conserved domain analysis and intron-exon distribution pattern of identified OsAP2s revealed their exclusive distribution among the specific clades of the phylogenetic tree. Moreover, the members of osa-miR172 family were also identified potentially targeting four OsAP2 genes. The real-time quantitative expression profiling of OsAP2s under heat stress conditions in contrasting indica rice genotypes revealed the differential expression pattern of OsAP2s (6 genes up-regulated and 4 genes down-regulated) in stress- and genotype-dependent manner. These findings unveiled the evolutionary pathways of AP2-TF in rice, and can help the functional characterization under developmental and stress responses.


Subject(s)
Evolution, Molecular , Heat-Shock Response , Oryza/genetics , Plant Proteins/genetics , Transcription Factors/genetics , Gene Duplication , Gene Expression Regulation, Plant , MicroRNAs/genetics , MicroRNAs/metabolism , Oryza/metabolism , Plant Proteins/metabolism , Selection, Genetic , Transcription Factors/metabolism
15.
Int J Mol Sci ; 23(21)2022 Oct 31.
Article in English | MEDLINE | ID: mdl-36362058

ABSTRACT

Cultivated soybean (Glycine max (L.)), the world's most important legume crop, has high-to-moderate salt sensitivity. Being the frontier for sensing and controlling solute transport, membrane proteins could be involved in cell signaling, osmoregulation, and stress-sensing mechanisms, but their roles in abiotic stresses are still largely unknown. By analyzing salt-induced membrane proteomic changes in the roots and leaves of salt-sensitive soybean cultivar (C08) seedlings germinated under NaCl, we detected 972 membrane proteins, with those present in both leaves and roots annotated as receptor kinases, calcium-sensing proteins, abscisic acid receptors, cation and anion channel proteins, proton pumps, amide and peptide transporters, and vesicle transport-related proteins etc. Endocytosis, linoleic acid metabolism, and fatty acid biosynthesis pathway-related proteins were enriched in roots whereas phagosome, spliceosome and soluble NSF attachment protein receptor (SNARE) interaction-related proteins were enriched in leaves. Using label-free quantitation, 129 differentially expressed membrane proteins were found in both tissues upon NaCl treatment. Additionally, the 140 NaCl-induced proteins identified in roots and 57 in leaves are vesicle-, mitochondrial-, and chloroplast-associated membrane proteins and those with functions related to ion transport, protein transport, ATP hydrolysis, protein folding, and receptor kinases, etc. Our proteomic results were verified against corresponding gene expression patterns from published C08 RNA-seq data, demonstrating the importance of solute transport and sensing in salt stress responses.


Subject(s)
Glycine max , Proteomics , Glycine max/genetics , Proteomics/methods , Membrane Proteins/metabolism , Sodium Chloride/pharmacology , Sodium Chloride/metabolism , Plant Roots/metabolism , Salt Stress , Plant Leaves/metabolism , Seedlings/genetics , Stress, Physiological/genetics , Plant Proteins/genetics , Plant Proteins/metabolism
16.
Int J Mol Sci ; 23(21)2022 Oct 26.
Article in English | MEDLINE | ID: mdl-36361700

ABSTRACT

Abiotic stresses, such as drought, salinity, heat, cold, and heavy metals, are associated with global climate change and hamper plant growth and development, affecting crop yields and quality. However, the negative effects of abiotic stresses can be mitigated through exogenous treatments using small biomolecules. For example, the foliar application of melatonin provides the following: it protects the photosynthetic apparatus; it increases the antioxidant defenses, osmoprotectant, and soluble sugar levels; it prevents tissue damage and reduces electrolyte leakage; it improves reactive oxygen species (ROS) scavenging; and it increases biomass, maintains the redox and ion homeostasis, and improves gaseous exchange. Glutathione spray upregulates the glyoxalase system, reduces methylglyoxal (MG) toxicity and oxidative stress, decreases hydrogen peroxide and malondialdehyde accumulation, improves the defense mechanisms, tissue repairs, and nitrogen fixation, and upregulates the phytochelatins. The exogenous application of proline enhances growth and other physiological characteristics, upregulates osmoprotection, protects the integrity of the plasma lemma, reduces lipid peroxidation, increases photosynthetic pigments, phenolic acids, flavonoids, and amino acids, and enhances stress tolerance, carbon fixation, and leaf nitrogen content. The foliar application of glycine betaine improves growth, upregulates osmoprotection and osmoregulation, increases relative water content, net photosynthetic rate, and catalase activity, decreases photorespiration, ion leakage, and lipid peroxidation, protects the oxygen-evolving complex, and prevents chlorosis. Chemical priming has various important advantages over transgenic technology as it is typically more affordable for farmers and safe for plants, people, and animals, while being considered environmentally acceptable. Chemical priming helps to improve the quality and quantity of the yield. This review summarizes and discusses how exogenous melatonin, glutathione, proline, and glycine betaine can help crops combat abiotic stresses.


Subject(s)
Melatonin , Melatonin/metabolism , Betaine/pharmacology , Betaine/metabolism , Proline/pharmacology , Proline/metabolism , Glutathione/metabolism , Antioxidants/pharmacology , Antioxidants/metabolism , Stress, Physiological/physiology
17.
Pak J Pharm Sci ; 35(1(Supplementary)): 259-265, 2022 Jan.
Article in English | MEDLINE | ID: mdl-35228186

ABSTRACT

Present study was aimed to formulate solid dispersions (SD) of Dapoxetine (DAPO), a drug with low water solubility with the help of PVP-30, Ploxomer-180 (P-180) and ß-cyclodextrin (CD) by physical mixing to enhance the water solubility and dissolution of drug. Binary and Ternary solid dispersions were prepared with different ratios of drug and polymer and were further evaluated the effects of polymer at individual level as well as when used in combination. The solid dispersions were whitish in color, irregular in shape and have some rough surface when illustrated under SEM. Percentage yield of SDs ranged from 81-95% described the method was structured and significant. Solubility studies were carried out in 0.1N Hcl solution and was observed a remarkable increase in solubility of BSD 8, BSD 12 and TSD 6. Furthermore, the formulations were characterized by FTIR and PXRD. P-180 found an exceptional solubility promoter when used alone and in combination with CD as well. A remarkable enhance in Solubility and dissolution of DAPO was investigated in formulation prepared with ternary SD of Pol-180 and CD.


Subject(s)
Benzylamines/chemistry , Drug Carriers/chemistry , Naphthalenes/chemistry , Polymers/chemistry , Selective Serotonin Reuptake Inhibitors/chemistry , beta-Cyclodextrins/chemistry , Microscopy, Electron, Scanning , Solubility , Spectroscopy, Fourier Transform Infrared
18.
Pak J Pharm Sci ; 35(4(Special)): 1191-1200, 2022 Jul.
Article in English | MEDLINE | ID: mdl-36218097

ABSTRACT

Farsetia hamiltonii Royle is a medicinal plant of Cholistan desert, Pakistan, traditionally used for treating diabetes, oxidative stress, arthritis, fever, gastrointestinal and respiratory diseases. This study represents unprecedented phytochemical, enzymatic and biological properties of F. hamiltonii root extracts to prove floric uses. Evaluation of Phytochemical constituents was done by screening, total flavonoid, phenolic contents and gas chromatography-mass spectrometry. The phytochemical screening revealed the presence of glycosides, bounded anthraquinones, flavonoids, saponins, steroids, coumarins and diterpenes in root extracts. Eight compounds were identified in dichloromethane extract, whereas one compound was identified in methanol extract of root part of F. hamiltonii. The dichloromethane extract possesses significant lipoxygenase, chymotripsin and cholinesterase enzyme inhibition activities, whereas methanol extract possess lipoxygenase, alpha glucosidase, chymotrypsin and acetylcholinesterase enzyme inhibition activities. The antibacterial activity of methanol extract was significant against selected five microbial strains. Nine compounds were reported in root part of F. hamiltonii first time. The enzyme inhibition assays on anti-cholinesterase, anti-alpha glucosidase, antilipoxygenase, antichymotripsin and antibacterial activities were found significant for the extracts of root parts of F. hamiltonii. Therefore, results of this study justify folkloric therapeutic potential of F. hamiltonii root in treating diabetes, inflammations and infectious diseases.


Subject(s)
Brassicaceae , Saponins , Acetylcholinesterase , Anthraquinones , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Antioxidants/pharmacology , Cholinesterase Inhibitors/pharmacology , Chymotrypsin , Coumarins , Flavonoids/analysis , Glycosides , Lipoxygenases , Methanol , Methylene Chloride , Phytochemicals/pharmacology , Plant Extracts/chemistry , Plant Extracts/pharmacology , alpha-Glucosidases
19.
Pak J Pharm Sci ; 35(6): 1549-1556, 2022 Nov.
Article in English | MEDLINE | ID: mdl-36789814

ABSTRACT

Plants are always better choice for treatment of disease in contrast to synthetic agents due to their less toxicity, and free availability with minor limitations of identifications, purity and potency which should be addressed. The objective of current study was to explore the anti-inflammatory effect of methanol extract of Yucca elephantipes roots by using oxidative burst assay and carrageen an induced rat paw edema. GC-MS analysis was carried for the determination of ani-inflammatory potential of fatty acids and other phytochemicals present in Yucca elephantipes roots. Among fifteen detected compounds trans-13-octadecenoic acid, n-hexadecanoic acid and 4-hydroxy benzoic acid were found as 84.21%, 5.21% and 2.17% respectively. Oxidative burst assay showed anti-inflammatory potential of Yucca elephantipes roots 74.58±0.32% with IC50 of 15.3±2.2µg/mL as compared to Ibuprofen with percentage of inhibition 73.20±0.17% and IC50 was 11.2±0.98µg/mL. Fortunately, less than 8g/kg dose of the Yucca elephantipes roots found safe in albino rats. Interestingly, in-vivo carrageen an induced paw edema method proved its anti-inflammatory potential at dose 100 and 200mg/kg in albino rats. Conclusively, 200mg/kg dose of Yucca elephantipes roots extract was optimized for 88.89±0.015% anti-inflammatory effect which can be considered most potent, safe and better alternative of synthetic drugs.


Subject(s)
Plant Extracts , Yucca , Anti-Inflammatory Agents/therapeutic use , Edema/chemically induced , Edema/drug therapy , Plant Extracts/chemistry , Yucca/chemistry , Animals , Rats
20.
Pak J Pharm Sci ; 35(4(Special)): 1261-1267, 2022 Jul.
Article in English | MEDLINE | ID: mdl-36218105

ABSTRACT

The purpose of this study was to purify the phytoconstituents and to explore the antibacterial, antifungal, phytotoxic and cytotoxic potential of dichloromethane and methanol extracts of aerial and root parts of Trigonella polycerata. The phytochemical study on methanol extract of aerial parts of the plant led to the isolation and purification of seven compounds that were identified as 3,4-dimethoxycinnamaldehyde, Trigocoumarin, 6,7,8-trimethoxycoumarin, Penduletin, 5-hydroxy-3,6,7,4´-tetramethoxyflavone, 3,5,7-trihydroxy-6,4-dimethoxyflavone and 5-hydroxy-4,7-dimethoxyflavone. These structures were elucidated by interpretation of EI-MS and NMR spectral data. The plant aerial parts methanol extract (TPAM) demonstrated higher antibacterial (78.99%), phytotoxic (85% growth regulation at 1000µg/mL) and cytotoxic activities (LD50: 45.643µg/mL). While the methanol root extract (TPRM) was highly active against bacteria's; Salmonella typhi (71.56%), Staphylococcus aureus (70.15%), Escherichia coli (69%), fungi like Candida albicans (70.21%) and moderately active against Brine shrimp larvae (LD50: 125.663µg/mL). The dichloromethane aerial (TPAD) and root (TPRD) extracts exhibited significant antibacterial (78.03% and 50.21% inhibitions respectively) and phytotoxic (55% growth regulation at 1000µg/mL) potential. Only TPAD indicated the best inhibition against fungi; Aspergillus flavus (75.31%) and moderate inhibition against Microsporum canis (42.21%). This phytochemical and biological work is the first time reported in Trigonella polycerata.


Subject(s)
Trigonella , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/toxicity , Antifungal Agents/chemistry , Methanol , Methylene Chloride/analysis , Microbial Sensitivity Tests , Phytochemicals/analysis , Phytochemicals/pharmacology , Plant Components, Aerial/chemistry , Plant Extracts/toxicity
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