Plant Cells Research Articles

Systematic multistep extraction process for the total valorization of fiber fractions from fruit seeds

This study explores a systematic multistep extraction process to valorize various fruit seeds (mango, cherry, lemon, pumpkin, avocado, litchi, peach, and apricot) by fractionating and isolating different fiber components. A first mild treatment, i.e., a protease-assisted extraction, was employed to disaggregate the plant cell structure and isolate soluble fibers. Subsequently, the insoluble residue underwent extraction by hot acidic water to solubilize and separate the pectin fraction. The remaining solid material was finally subjected to hydrothermal treatment to recover recalcitrant hemicellulose. Cherry, lemon, and pumpkin seeds emerged as the most advantageous by-products overall, showing great extractable quantities of low molecular weight pectin, pectin oligosaccharides, arabinogalactans, xylan, and xylo-oligosaccharides (extraction yields up to 63% and 68% in the first and third step, respectively). Peach and apricot seeds yielded fiber extracts that were similar to each other but distinct for each fraction, suggesting potential applications in different food formulations, albeit with lower yields. The purity of the extracts increased with each stage, reaching 42.6% for soluble fibers, 67.3% for pectin, and 76.1% for hemicellulose. Mango, avocado, and litchi seeds were less suitable for this method but could be utilized for starch recovery. This cascade process effectively maximizes the utilization of fruit by-products, offering a scalable solution for the sustainable extraction of valuable fibers.

Arabidopsis F-box proteins D5BF1 and D5BF2 negatively regulate Agrobacterium-mediated transformation andtumorigenesis.

The pathogen Agrobacterium tumefaciens is known for causing crown gall tumours in plants. However, it has also been harnessed as a valuable tool for plant genetic transformation. Apart from the T-DNA, Agrobacterium also delivers at least five virulence proteins into the host plant cells, which are required for an efficient infection. One of these virulence proteins is VirD5. F-box proteins, encoded in the host plant genome or the Ti plasmid, and the ubiquitin/26S proteasome system (UPS) also play an important role in facilitating Agrobacterium infection. Our study identified two Arabidopsis F-box proteins, D5BF1 and D5BF2, that bind VirD5 and facilitate its degradation via the UPS. Additionally, we found that Agrobacterium partially suppresses the expression of D5BF1 and D5BF2. Lastly, stable transformation and tumorigenesis efficiency assays revealed that D5BF1 and D5BF2 negatively regulate the Agrobacterium infection process, showing that the plant F-box proteins and UPS play a role in defending against Agrobacterium infection.

Barley as a production platform for oral vaccines in sustainable fish aquaculture

Vaccination is the most effective measure to prevent disease outbreaks in fish aquaculture, with oral vaccine administration emerging as the most practical approach. However, oral vaccines face a notable limitation due to insufficient stimulation of the complex gut-associated lymphoid tissue caused by factors such as vaccine degradation, poor absorption, and recognition by the immune cells. An innovative solution to these limitations lies in the plant-based production of recombinant vaccines. Plant cells enable the production and targeted storage of recombinant vaccines in specific cell organelles which ensure superior protection from degradation and contain natural compounds acting as adjuvants. Our study explores the potential of barley (Hordeum vulgare), a globally significant cereal crop, for producing orally administered subunit vaccines against viral infections affecting economically important fish species in the Salmonidae and Cyprinidae families. Through Agrobacterium-mediated transformation of immature barley embryos, we have generated homozygous T2 generation of transgenic barley expressing recombinant antigens of spring viremia of carp virus and infectious salmon anaemia virus. The expression of these plant-based recombinant vaccines was confirmed by immunodetection, which was supported by fluorescence observation, specifically in the seed endosperm. The antigenicity of transgenic plant material containing recombinant antigens was evaluated using an intubation model of common carp (Cyprinus carpio), revealing a substantial upregulation of the immunoglobulin transcripts in both systemic and mucosal tissues over a period of 28 days following a single dose of transgenic antigens. Collectively, these results underscore the potential of barley-based recombinant vaccines for disease prevention in fish aquaculture.

Homologous expression, purification, and characterization of a recombinant acetylxylan esterase from Aspergillus nidulans

Acetylxylan esterases (AXEs) are essential enzymes that break down the acetyl groups in acetylated xylan found in plant cell walls polysaccharides. They work synergistically with backbone-depolymerizing xylanolytic enzymes to accelerate the degradation of complex polysaccharides. In this study, we cloned the gene axeA, which encodes the acetylxylan esterase from Aspergillus nidulans FGSC A4 (AxeAN), into the pEXPYR expression vector and introduced it into the high protein-producing strain A. nidulans A773. The purified AxeAN, with a molecular weight of 33.5 kDa as confirmed by SDS-PAGE, was found to be active on ρ-nitrophenyl acetate (ρNPA), exhibiting a remarkably high specific activity (170 U mg−1) at pH 7.0 and 55 °C. AxeAN demonstrated stability over a wide pH range (5.5–9.0), retaining >80% of its initial activity after 24 h. The KM and Vmax were 0.098 mmol L−1 and 320 U mg−1, respectively, using ρNPA as a substrate. We also evaluated the synergistic effect of AxeAN with an endo-1,4-β-xylanase from Malbranchea pulchella (MpXyn10) in the hydrolysis of four different xylans (Birchwood, Beechwood, Oat spelt, and Arabinoxylan) to produce xylooligosaccharides (XOS). The best results were obtained using Birchwood xylan as substrate and MpXyn10-AxeAN as biocatalysts after 24 h of reaction (50 °C), with a XOS-yield of 91%, value 41% higher when compared to MpXyn10 (XOS-yield of 63%). These findings showed the potential of the application of AxeAN, together with other xylanases, to produce xylooligosaccharides with high purity and other products with high added value in the field of lignocellulosic biorefinery.

Sucrose-responsive osmoregulation of plant cell size by a long non-coding RNA

In plants, sugars are the key source of energy and metabolic building blocks. The systemic transport of sugars is essential for plant growth and morphogenesis. Plants evolved intricate molecular networks to effectively distribute sugars. The dynamic distribution of these osmotically active compounds is a handy tool for regulating cell turgor pressure, an instructive force in developmental biology. Here, we set out to investigate the molecular mechanism behind the dual role of a receptor-like kinase CANAR. We functionally characterized a long non-coding RNA, CARMA, as a negative regulator of CANAR. Sugar-responsive CARMA specifically fine-tunes CANAR expression in the phloem, the route of sugar transport. Based on our genetics, molecular, microscopy, and biophysical data, we propose that by controlling sugar phloem transport from shoot to root, the CARMA-CANAR module allows cells to flexibly adapt to the external osmolality by appropriate water uptake and thus adjust the size of vascular cell types during organ growth and development. We identify a nexus of plant vascular tissue formation with cell internal pressure monitoring and reveal a novel functional aspect of long non-coding RNAs in developmental biology.

Application of Hazard Analysis Critical Control Point (HACCP) principles to beef mortadella production

A mortadella is a cooked sausage made from raw pork, beef, chicken or lamb. It is popular in Jordan and neighboring countries for its taste, texture, nutritional value and ease of inclusion in sandwiches. The product is sold in plastic packaging, in circular rolls or vacuum packed. Although it poses no serious threat to human health, the heat can be dangerous if used improperly. A HACCP system is used to manage the production process, with a focus on quality control (CCP) throughout the manufacturing phase. Safety risks are identified at all stages of production, from raw materials to storage and distribution. Foodborne illnesses are primarily caused by pathogenic microorganisms, and these raw materials include pathogenic plant cells, Bacillus cereus, Clostridium botulinum and Clostridium perfringens that survive after cooking and contamination detected after monitoring. Post-manufacturing controls are performed through Good Manufacturing Practices (GMPs). The objectives of the study were to identify potential hazards associated with the production of beef mortadella, determine the CCPs to be implemented, develop a monitoring plan and establish corrective action on a case-by-case basis. The HACCP plan was developed in collaboration with local stakeholders and follows a process flow diagram to identify potential hazards and preventive measures. The system includes the steps for obtaining and storing the ingredients, preparing the meat, adding non-meat ingredients, and transferring the raw mixture to an incubator. The meat is weighed and filled with the mixture to improve cleanliness and avoid excess nitrite. The storage time between incubation and cooking is controlled to allow bacteria to multiply. To kill plant pathogens and reduce the total number of microorganisms, the product is cooked in a steam oven at 75°C for 5 minutes. The aim of the cooking process is to reduce microbial growth before lethal temperatures have been reached and uniform heating of the product has been ensured. Cooking suitability is checked by thermographic records and visual inspection of the cooked product. The plan also includes personnel in charge of every stage of production, ensuring a clean and hygienic environment. Key words: HACCP, Mortadella, meat processing, hygiene control, food safety

Investigating Removal of Carbamazepine by Helianthus annuus Plant Cells

Investigating Removal of Carbamazepine by Helianthus annuus Plant Cells

Lignin-Based Composite Film and Its Application for Agricultural Mulching.

Agricultural mulching is an important input for modern agricultural production and plays an important role in guaranteeing food security worldwide. At present, polyethylene (PE) mulching is still commonly used in agricultural production in most countries around the world, which is non-biodegradable, and years of mulching have caused serious agricultural white pollution. Lignin is one of the three major components of plant cell walls, and it is also the main renewable natural aromatic compounds in nature. Lignin-based composite film materials are green, biodegradable, and show good prospects for development in the field of agricultural mulch. This paper introduces the types, structure, and application status of lignin, summarizes the preparation of lignin-based composite film materials and its latest research progress, focuses on the types, preparation methods, and application examples of lignin-based agricultural mulching, and looks forward to the future development prospects of lignin-based agricultural mulching.

Effects of biomass feedstock and applied pressure on the binding mechanism and pellet qualities

To expand the sources of feedstocks for pelleting and enhance the applicability of the pelleting device, 7 types of feedstocks with representative chemical component contents from different resources were densified. The pressure-time (P-T) curves and energy consumption were used to analyze the densification process and binding mechanism. Specific energy density and densification effects were evaluated pellet qualities and classified feedstock types. Results showed that the densification process was first affected by the feedstock types and then applied pressure. Biomass chemical component contents significantly influence the densification process, the required time, and the final pellet length. Energy consumption in the compression stage is much more than in the extrusion stage. The exponential stage consumes more than 90 % of the total energy. Energy conversion efficiency varies from 62.19 % to 95.06 % with the feedstock types and applied pressures. Single pellet density, densification effects, and specific energy densities increased with the rising applied pressure. BB, SD, PN, CS, and SB can be used as primary feedstocks. SR and ROC act as secondary feedstocks or lubricants. Cellulose, hemicellulose, and lignin of plant cell walls are solid bridges. Extractives, fat, and protein in the plant cells are binders and lubricants to make the pellet surface smooth. This paper could provide further insight into various biomass feedstock densification and general design for pelleting devices.

Surface-enhanced Raman spectroscopy for the characterization of xylanases enzyme

Xylanases are essential hydrolytic enzymes which break down the plant cell wall polysaccharide, xylan composed of D-xylose monomers. Surface-enhanced Raman Spectroscopy (SERS) was utilized for the characterization of interaction of xylanases with xylan at varying concentrations. The study focuses on the application of SERS for the characterization of enzymatic activity of xylanases causing hydrolysis of Xylan substrate with increase in its concentration which is substrate for this enzyme in the range of 0.2% to 1.0%. SERS differentiating features are identified which can be associated with xylanases treated with different concentrations of xylan. SERS measurements were performed using silver nanoparticles as SERS substrate to amplify Raman signal intensity for the characterization of xylan treated with xylanases. Principal Component Analysis (PCA) and Partial Least Square Discriminant Analysis (PLS-DA) were applied to analyze the spectral data to analyze differentiation between the SERS spectra of different samples. Mean SERS spectra revealed significant differences in spectral features particularly related to carbohydrate skeletal mode and O-C-O and C–C–C ring deformations. PCA scatter plot effectively differentiates data sets, demonstrating SERS ability to distinguish treated xylanases samples and the PC-loadings plot highlights the variables responsible for differentiation. PLS-DA was employed as a quantitative classification model for treated xylanase enzymes with increasing concentrations of xylan. The values of sensitivity, specificity, and accuracy were found to be 0.98%, 0.99%, and 100% respectively. Moreover, the AUC value was found to be 0.9947 which signifies the excellent performance of PLS-DA model. SERS combined with multivariate techniques, effectively characterized and differentiated xylanase samples as a result of interaction with different concentrations of the Xylan substrate. The identified SERS features can help to characterize xylanases treated with various concentrations of xylan with promising applications in the bio-processing and biotechnology industries.

Evaluation of a 16S RRNA Amplicon Illumina Sequencing Method for Examining the Gut Microbiome of <i>Lipaphis erysimi</i> (Kalt)

Herbivore specialists adapt to feed on a specific group of host plants by evolving various mechanisms to respond to plant defenses. Gene enrichment in the metagenome, accompanied by functional identification, revealed an essential role of specific gut bacteria in the breakdown of plant cell walls, detoxification of plant phenolics, and synthesis of amino acids. The Lipaphis erysimi (Kalt) gut bacteriome is an essential aspect for herbivory adaptation and this study is aimed at the microbiome of the musatrd Aphid L. erysimi gut by targeting the V3-V4 hyper variable region of the 16S rRNA gene with the Illumina platform sequencing using adult L. erysimi was performed to understand the bacteriome variations. A total of 103883 reads were obtained revealing the gut of the L. erysimi was found to be dominated by the main symbiont Buchnera a Protobacteria.

Functional and evolutionary insights into chemosensation and specialized herbivory from the genome of the red milkweed beetle, Tetraopes tetrophthalmus (Cerambycidae: Lamiinae).

Tetraopes are aposematic longhorn beetles (Cerambycidae) that feed primarily on toxic plants in the genus Asclepias (milkweeds). Studies of Tetraopes and their host plants have revealed compelling evidence for insect-plant coevolution and cospeciation. We sequenced, assembled and annotated the genome of the common red milkweed beetle, Tetraopes tetrophthalmus, and explored gene content and evolution, focusing on annotated genes putatively involved in chemosensation, allelochemical detoxification, and phytophagy. Comparisons were made to the Asian longhorned beetle (Anoplophora glabripennis) genome. The genome assembly comprised 779Mb distributed across 1057 contigs, with an N50 of 2.21Mb and 13,089 putative genes, including 97.3% of expected single-copy orthologs. Manual curation identified 122 putative odorant receptors (OR) and 162 gustatory receptors (GR), the former number similar to A. glabripennis but the latter only 69% of the A. glabripennis suite. We also documented a greater percentage of pseudogenic GRs and ORs compared to A. glabripennis, suggesting an ongoing reduction in chemosensory function, perhaps related to host specialization. We found lower diversity within certain well-studied gene families predicted to encode putative plant cell wall degrading enzymes in the T. tetrophthalmus genome, perhaps also due to host specialization. Exploring genes relevant to stress and allelochemical detoxification revealed evidence of an abundance of ABC-family genes in the T. tetrophthalmus genome, which may be related to sequestering toxic cardiac glycosides. Our studies further illuminate the genomic basis and evolution of chemosensation in longhorn beetles and provide a new vantage point from which to explore the ecology and evolution of specialized plant-feeding in Tetraopes and other phytophagous beetles.

185th anniversary of the cell theory and modern cell technologies

The article briefly reviews the history of the development of the cell theory in the 19th century and analyses the contribution of its authors Theodor Schwann, Matthias Schleiden, and Rudolf Virchow. Particular attention is paid to the development of the cell theory and its practical significance, in particular, the invention in the 20th century and further development of cell technologies. The main characteristics of cultured cells and tissues in vitro as a new experimentally created biological system are presented, and the peculiarities of the establishment of strains of cultured plant cells are discussed. Using plants as an example, the genomic variability of somatic cells is analysed both during ontogeny and in the course of their culturing in vitro. It is assumed that any somatic cell with a living (functionally active) nucleus, when isolated and subsequently cultured in vitro can restore in its offspring, including regenerated plants, the genetic polymorphism inherent in this plant species as a result of dedifferentiation and “somaclonal” variability (the latter is assumed to occur according to the N. I. Vavilov’s law of homologous series in variation). It ispostulated that a plant is a system of cell populations characterised by the plasticity of its gene pool, which is based on genome plasticity of somatic cells, which, in interaction with cellular selection, ensures the adaptability of the plant as an integral organism and creates the possibility of inheritance (transmission to offspring) of adaptive genomic changes acquired during ontogeny. It is concluded that the adaptive traits of an organism are determined by the adaptive changes in the cells, which composed it. The author also examines the main directions of modern cell technologies and their application in crop production, in medicine for the treatment and creation of new organs using stem cells, as well as in the food industry on the example of the development of technologies for the production of artificial meat using animal stem cells, etc. It is concluded that the issue of cell adaptations remains one of the most important and still insufficiently understood issues of modern biology.

Dual regulation of thermal conductivity and mechanical performance of nano cellulose-based composite via mimicking plant cell wall structure

Combining thermal conductive fillers and flexible polymers is an agile approach to fabricating composites with heat-conducting performance. However, the thermal conductivity of the composites is hard to reach an equal level to the functional fillers. The mainspring is that the thermally conductive pathways within the composite could not be well-constructed due to the air-induced interface thermal resistance. Herein, inspired by the plant cell wall structure, polyvinyl alcohol (PVA) with abundant hydroxyl groups was adopted as a binder for boosting the thermally conductive pathways construction between cellulose nanofiber (CNF) and alkalized hexagonal boron nitride (BN-OH), also for strengthening the mechanical performance of the composite. The results showed that the tensile strength and through-plane thermal conductivity of the composite were high up to 91.0 MPa and 2.2 W m−1 K−1 at 40 wt% PVA content, exhibiting 121 % and 450 % enhancements compared to pure CNF film (41.2 MPa and 0.4 W m−1 K−1). Moreover, the composite also presented high thermal stability (decomposition temperature of onset was 218 °C) and good hydrophobicity properties. Overall, this study innovatively proposes an idea for enhancing the thermal conductivity and improving the mechanical properties of the composite, which is indispensable for developing thermal management materials for next-generation electronics.

Progressive heterogeneity of enlarged and irregularly shaped apicoplasts in P. falciparum persister blood stages after drug treatment.

Morphological modifications and shifts in organelle relationships are hallmarks of dormancy in eukaryotic cells. Communications between altered mitochondria and nuclei are associated with metabolic quiescence of cancer cells that can survive chemotherapy. In plants, changes in the pathways between nuclei, mitochondria, and chloroplasts are associated with cold stress and bud dormancy. Plasmodium falciparum parasites, the deadliest agent of malaria in humans, contain a chloroplast-like organelle (apicoplast) derived from an ancient photosynthetic symbiont. Antimalarial treatments can fail because a small fraction of the blood stage parasites enter dormancy and recrudesce after drug exposure. Altered mitochondrial-nuclear interactions in these persisters have been described for P. falciparum, but interactions of the apicoplast remained to be characterized. In the present study, we examined the apicoplasts of persisters obtained after exposure to dihydroartemisinin (a first-line antimalarial drug) followed by sorbitol treatment, or after exposure to sorbitol treatment alone. As previously observed, the mitochondrion of persisters was consistently enlarged and in close association with the nucleus. In contrast, the apicoplast varied from compact and oblate, like those of active ring stage parasites, to enlarged and irregularly shaped. Enlarged apicoplasts became more prevalent later in dormancy, but regular size apicoplasts subsequently predominated in actively replicating recrudescent parasites. All three organelles, nucleus, mitochondrion, and apicoplast, became closer during dormancy. Understanding their relationships in erythrocytic-stage persisters may lead to new strategies to prevent recrudescences and protect the future of malaria chemotherapy.

How Many Glucan Chains Form Plant Cellulose Microfibrils? A Mini Review.

Assessing the number of glucan chains in cellulose microfibrils (CMFs) is crucial for understanding their structure-property relationships and interactions within plant cell walls. This Review examines the conclusions and limitations of the major experimental techniques that have provided insights into this question. Small-angle X-ray and neutron scattering data predominantly support an 18-chain model, although analysis is complicated by factors such as fibril coalescence and matrix polysaccharide associations. Solid-state nuclear magnetic resonance (NMR) spectroscopy allows the estimation of the CMF width from the ratio of interior to surface glucose residues. However, there is uncertainty in the assignment of NMR spectral peaks to surface or interior chains. Freeze-fracture transmission electron microscopy images show cellulose synthase complexes to be "rosettes" of six lobes each consistent with a trimer of cellulose synthase enzymes, consistent with the synthesis of 18 parallel glucan chains in the CMF. Nevertheless, the number of chains in CMFs remains to be conclusively demonstrated.

Regulating PCD in plant cells: Intracellular acidification plays a pivotal role together with calcium signaling.

Programmed cell death (PCD) occurs in different tissues in response to a number of different signals in plant cells. Drawing from work in several different contexts, including root cap cell differentiation, plant response to biotic and abiotic stress, and some self-incompatibility (SI) systems, the data suggest that, despite differences, there are underlying commonalities in early decision-making stages of PCD. Here, we focus on how two cellular events, increased [Ca2+]cyt levels and cytosolic acidification, appear to act as early signals involved in regulating both developmental- and stimulus-induced PCD in plant cells.

Climate-adaptive battery solutions for renewable microgrids: A case study in Indian coastal and inland regions

The utilization of renewable energy has the potential to alleviate global warming, reduce carbon emissions in the environment, and offer sustainable energy solutions to remote regions. Presently, 13 % of the worldwide population resides in isolated inland and coastal areas where electricity remains inaccessible. Consequently, implementing microgrids powered by renewable energy sources becomes essential to enhance electricity accessibility in these remote locations. This study aims to identify efficient and cost-effective renewable energy technologies suitable for deployment inland and coastal areas. The techno-economic feasibility of renewable energy systems is being evaluated in two distinct locations: Yadavole (inland) and Uppada (coastal) villages in the Indian state of Andhra Pradesh. The energy analysis encompasses various sources, including photovoltaic, diesel generators, wind turbines, flywheels, and batteries (Li-ion and lead-acid). The HOMER software conducts all necessary modeling and simulations for economic analysis and optimal system sizing. According to the simulation results, the most viable microgrid configurations for the inland region, Yadavole, are photovoltaic/diesel and generator/Li-ion. In contrast, for the coastal region, Uppada, photovoltaic/diesel, and generator/lead-acid configurations prove to be the most promising configurations. Moreover, a MATLAB model of the proposed system is created to analyze energy quality. Given the superior performance of Li-ion batteries in elevated temperatures typically found inland, this study recommends a diversified range of battery solutions. Specifically, Li-ion batteries are recommended for inland areas, while lead-acid batteries are recommended for coastal regions. This approach aims to efficiently store renewable power by the specific climatic conditions of each area. It is crucial to systematically analyze region-specific renewable energy technologies to provide valuable insight to stakeholders involved in investment and energy policy decisions.

Genetic Evidence of SpGH9A3 in Leaf Morphology Variation of Spathiphyllum 'Mojo'.

Leaves play a crucial role as ornamental organs in Spathiphyllum, exhibiting distinct differences across various Spathiphyllum varieties. Leaf development is intricately linked to processes of cell proliferation and expansion, with cell morphology often regulated by plant cell walls, primarily composed of cellulose. Alterations in cellulose content can impact cell morphology, subsequently influencing the overall shape of plant organs. Although cellulases have been shown to affect cellulose levels in plant cells, genetic evidence linking them to the regulation of leaf shape remains limited. This study took the leaves of Spathiphyllum 'Mojo' and its somatic variants as the research objects. We screened four cellulase gene family members from the transcriptome and then measured the leaf cellulose content, cellulase activity, and expression levels of cellulase-related genes. Correlation analysis pinpointed the gene SpGH9A3 as closely associated with leaf shape variations in the mutant. Green fluorescent fusion protein assays revealed that the SpGH9A3 protein was localized to the cell membrane. Notably, the expression of the SpGH9A3 gene in mutant leaves peaked during the early spread stage, resulting in smaller overall leaf size and reduced cellulose content upon overexpression in Arabidopsis.

Drought stress enhances phytochemical, antioxidant, antidiabetic and anticancer properties in Pandanus tectorius

Global warming is responsible for increasing temperatures and reducing rainfall in many regions of the world. Due to insufficient rainfall or lack of water in the soil, plants experience a range of morphological and biochemical alterations when they are exposed to drought conditions. Interestingly, the cells of drought-stressed plants can serve as a potential source for discovering new drugs. These secondary metabolic products have important biological functions that determine how plants interact with the environment and help them accommodate alterations. Present research has shed light on the response of Pandanus tectorius induced drought stress at 8th and 16th days. The studies observed the levels of various secondary metabolites in plants under drought-induced stress. Among the secondary metabolites analyzed, total alkaloids exhibited the highest overall concentration at 26.19 ± 0.83a. Furthermore, the observations found that drought stress had a significant impact on the plant's secondary metabolism. This resulted in increased levels of total alkaloids, saponins, tannins, and phenolics compared to the control plant. Especially the concentration of phenolics showed the most significant increase, with a remarkable 64.99% rise observed at the 16th day of treatment. The concentration of phenolics reached (20.52 ± 0.181c) at this level. Moreover, compared to control, P. tectorius exhibits significant antioxidant, antidiabetic, anti-inflammatory, molecular docking, and anticancer effects against MDA-MB-231 human breast cancer cell line inhibitory concentration IC50 values (48.91 ± 0.95 μg/mL) at 16th day treated plants. Consequently, the positive responses of P. tectorius drought stress enhance secondary metabolites. This research focus on drought stress stimulus prompts an urge in the production of secondary metabolites in P. tectorius, leading to an increase in its anticancer and anti-diabetic properties and treatments for type-2 diabetes and against fighting for cancer with minimal side effects and helping prevent diseases.