Viral diseases, representing the most frequent emerging infectious diseases in plants by causing significant economic losses in agricultural production. Investigating tripartite interactions among plants, pathogens and biological resistance inducers is essential for understanding plant immune system. In plant-virus interactions, resistance often depends on the fast upregulation of defense responses. Several molecular pathways and specific transcription factors (TFs) were regulated, leading to gene expression changes that result in the synthesis of effector proteins and metabolites conferring resistance against viral diseases. Upon virus detection, multiple signaling cascades are activated, ultimately causing transcriptional reprogramming in plant cells. This process is modulated by various TFs, including the WRKY family that are involved in defense mechanisms. This family has been identified across multiple plant species. In this review we examine the role of the WRKY gene family in regulating plant defense responses against viral pathogens.
Nanoparticles offer promising applications in agriculture due to their unique physicochemical properties. This study investigated selenium nanoparticles (SeNPs) as a rooting agent to improve rice tolerance to low phosphate stress. IR64 rice was grown in media with varying SeNP concentrations (0-25 ppm), and plants were analyzed at 2 and 6 weeks for phenotypic traits, histology, biochemistry, and gene expression. At 2 weeks, 25 ppm SeNPs led to a threefold increase in root length versus untreated controls. Histological analysis revealed increased root perimeter and diameter, along with reduced aerenchyma perimeter. After 6 weeks under phosphate starvation, SeNP-treated plants showed higher phosphate and selenium content in roots and shoots, increased shoot length and weight, and reduced root length, weight, and phenolic content. Gene expression analysis showed that SeNPs upregulated key phosphate deficiency response genes, including OsPAP21, OsPT9, OsSPX, and OsPHR2. The most dramatic change was observed in OsSPX expression, which increased nearly 250-fold in shoots under full phosphate conditions. This is the first study to demonstrate that SeNPs promote root growth, enhance phosphate uptake, and regulate gene expression, suggesting SeNPs may serve as a sustainable strategy to boost phosphate use efficiency in rice cultivation.
The terpene synthase (TPS) gene family is integral to the biosynthesis of terpenoids, which are vital for plant defence, development, and interaction with the environment. Yellowhorn (Xanthoceras sorbifolium) has gained attention for its bioactive compounds, particularly terpenoids, which have applications in pharmaceuticals, biofuels, and cosmetics. This study provides a comprehensive pan-genome-wide analysis of the TPS gene family across five yellowhorn varieties (Xg11, Xzs4, Xwf8, Xjg, and Xzg2). A total of 257 TPS genes were identified and characterised, showing diversity in their evolutionary patterns. Phylogenetic analysis revealed distinct clades corresponding to functional classes of TPS genes. Conserved domains and motifs of these genes were analysed to highlight their structural characteristics. Furthermore, expression profiling under abiotic stresses, including cold and drought, was conducted, revealing the roles of specific TPS genes in stress tolerance. Tissue-specific expression analysis demonstrated the involvement of TPS genes in key physiological processes across different plant organs. This research advances our understanding of the TPS gene family in yellowhorn, with implications for improving crop resilience and biotechnological applications.
The rubber tree (Hevea brasiliensis) is highly valued for its wood and latex. Along with being a rich source of polyterpenes in the latex, rubber trees are notable for their significant emission of volatile monoterpenes that may contribute to its chemical defense. A previous in silico genome-wide analysis of Hevea brasiliensis identified 47 putative terpene synthase (HbTPS) genes, of which only one had been experimentally characterized. In this study, we report the isolation and functional characterization of HbTPS24, a new monoterpene synthase belonging to the HbTPS family, providing new insights into the molecular basis of monoterpene biosynthesis in this species. Phylogenetic analysis placed HbTPS24 in the TPS-b subfamily. In vitro enzymatic assays revealed that HbTPS24 is multi-functional, converting both neryl diphosphate and geranyl diphosphate into cyclic monoterpenes such as α-thujene, β-pinene, α-terpinene, α-terpinolene, and α-terpineol. qRT-PCR analysis showed that HbTPS24 is mainly expressed in the bark, with significantly lower levels in the leaves and undetectable levels in the latex. The predominant expression of HbTPS24 in bark tissues, the detection of specific monoterpenes such as D-limonene and α-pinene in bark extracts, along with the broader range of cyclic monoterpenes identified in both in vitro enzymatic assays and canopy-scale volatile flux measurements, strongly suggest that HbTPS24 contributes to the in planta synthesis of multiple cyclic monoterpenes. These compounds are potentially involved in mediating stress resilience in Hevea brasiliensis.
Global climate change poses a serious threat to agriculture, with heat and drought stress often occurring simultaneously and severely impacting crop productivity. As post-transcriptional regulators, microRNAs (miRNAs) mediate plant responses to these adverse conditions by targeting genes involved in antioxidant defense, growth, development, and hormonal signaling. However, research on miRNA roles under combined drought and heat stress is still limited compared to individual stress studies. Additionally, stress-, cultivar-, and tissue-specific expression patterns of miRNAs, along with discrepancies between controlled laboratory conditions and natural environments, complicate the development of broadly applicable miRNA-based strategies. This review explores recent advancements in understanding miRNA target genes and their functions, highlighting the need for innovative, sustainable solutions for crop improvement.
Drought is one significant environmental stress that has a detrimental impact on plant growth and lowers production. This study examined the role of ZnO₂ nanoparticles (NPs) in reducing the harmful effects of drought. The growth features of marjoram plants cultivated in typical and drought-stressed environments (100, 75, and 50% water field capacity) were increased by spraying with solutions containing nanoparticles of ZnO2. In contrast to other treated plants, plants treated with ZnO2 NPs (100 mg L-1) showed greater levels of important osmolytes and conserved more photosynthetic pigments. Furthermore, by raising the membrane stability index, the antioxidant enzymes' activity, and the levels of lipid peroxidation, hydrogen peroxide, and exogenous application of ZnO2 NPs spraying improved drought tolerance and reduced the membrane injury index. An increase in growth and biochemical parameters, such as total antioxidant activity and anthocyanin content, was detected in ZnO2 NPs-treated drought-stressed marjoram plants. Marjoram plant oil analysis revealed that some components appeared, such as eucalyptol, and others increased, especially in response to treatment with only the lowest water stress (50% WFC) and 100 mg L-1 ZnO2 NPs treatment. Our findings generally corroborated the beneficial effects of applying zinc NPs, particularly the absolute contribution of ZnO2 NPs (100 mg L-1) to lessen drought stress's negative consequences (specifically, 50% WFC) in marjoram plants.
Agricultural productivity is increasingly constrained by water scarcity, which affects nearly one-quarter of cultivated land and is projected to intensify due to climate change and escalating freshwater demands. Melatonin is widely recognized as a potent biostimulant that plays a crucial role in mitigating various abiotic stresses, particularly drought, across many plant species. This study demonstrates that exogenous application of melatonin (150 μM; foliar spray) confers protection to Phaseolus vulgaris under moderate water deficit (40% field capacity). A randomized complete block design comprising four treatment groups (n = 30 seedlings per group, five replicates) was employed to systematically evaluate morphological, physiological, biochemical, and molecular responses. Melatonin applications at 21 and 28 days after sowing significantly enhanced shoot elongation, leaf area expansion, and photosynthetic efficiency. Biochemically, melatonin markedly increased the activities of key antioxidant enzymes (ascorbate peroxidase (APX), catalase (CAT), and peroxidase (POD)), reduced reactive oxygen species accumulation, elevated proline content by 24%, and decreased electrolyte leakage by 18%, thereby improving osmotic balance and maintaining membrane integrity. Genomic stability was assessed using Inter-Simple Sequence Repeat (ISSR) and Random Amplified Polymorphic DNA (RAPD) markers, revealing that melatonin substantially attenuated drought-induced DNA damage. Marker analysis further demonstrated differential sensitivity, and key statistical indices, including polymorphism information content (PIC), effective multiplex ratio (EMR), and resolving power (RP), exhibited strong linear associations, reinforcing the reliability of molecular diagnostics. Collectively, these results highlight melatonin's multifaceted role in enhancing water-deficit resilience through integrated regulation of physiological homeostasis, oxidative stress mitigation, and genome protection. The findings support melatonin's practical potential as a low-cost, environmentally compatible strategy for improving legume performance in water-deficit environments.
Powdery mildew, caused by Erysiphe necator, is a major fungal disease affecting grapevines, leading to significant yield and quality losses. Plants defend against pathogens through pattern-triggered immunity (PTI) and effector-triggered immunity (ETI), with nucleotide-binding leucine-rich repeat receptors (NLRs) playing a crucial role in ETI. Hormonal signaling pathways, including salicylic acid (SA), jasmonic acid (JA), and ethylene (ET), further regulate immune responses. This study functionally characterizes two NLR genes (XP_010660247.1 and XP_003633889.1) responsive to powdery mildew in Arabidopsis thaliana. Using transgenic plants overexpressing these genes individually and in combination, we analyzed their expression patterns in response to E. necator infection and various phytohormones. Real time analysis demonstrated varied tissue-specific expression profiles across all transgenic lines. XP_010660247.1 lines exhibited early and sustained immune response post E. necator infection, indicating robust activation of the SA pathway. XP_003633889.1 lines showed delayed but prolonged activation of SA pathway genes, suggesting nuanced temporal dynamics in fungal resistance. Stacked NLR lines showed further delay in activation of SA pathway genes possibly due to mutual cross-interference in gene activation. Enhanced JA/SA pathway activation indicates heightened defense against biotic stress response. Crosstalk between JA and abscisic acid (ABA) pathways was evidenced by MYC2 upregulation in response to methyl jasmonate and ABA treatments. The ET pathway demonstrated nuanced activation with sensitivity to ET elicitor 1-aminocyclopropane-1-carboxylic acid but the inability to further activate the ET pathway in transgenic lines. Our findings provide insights into plant immune mechanisms and offer potential strategies for engineering powdery mildew-resistant grapevine varieties.
Andrographis paniculata, commonly known as kalmegh is a highly valued medicinal plant. Pot-grown plants were subjected to water stress at vegetative, flowering, and fruiting stage by withholding water supply, followed by rewatering to facilitate recovery. Plants at the flowering and fruiting stage were particularly sensitive to drought stress compared to those at the vegetative stage. The plants were analysed for four diterpenoid compounds, namely andrographolide, 14-deoxyandrographolide, neoandrographolide, and andrograpanin. In plants subjected to stress at the vegetative and flowering stage, total andrographolide content increased significantly (P ≤ 0.05), by as much as 37% and 44%, respectively, over the levels in the control following 6 or more days of exposure, but remained unaffected in plants subjected to stress at the fruiting stage. Across all three stages, a significant decrease was observed in dry weight, relative water content (RWC), photosynthesis, conductance, and transpiration. Total andrographolide content was negatively correlated to dry weight, RWC, and rate of photosynthesis. These findings are useful in (1) identifying the ideal harvesting stage to achieve peak levels of bioactive compounds, (2) scheduling irrigation more efficiently to minimise yield loss due to water stress and maximise the content of bioactive compounds, and (3) developing stress-tolerant genotypes.
Global climate change is accompanied by an increase in the frequency and severity of droughts, which negatively impact plant growth. One of the protective mechanisms that allows plants to withstand the negative drought effects is the lignification of vascular and dermal tissues, which prevents water loss. Despite some progress in research on lignification in model and crop plants, the characteristics of lignin accumulation in conifers need further investigation. In this study, we examined the adaptation of Scots pine (Pinus sylvestris) seedlings to water deficit at weak (-0.15 MPa) and strong (-0.5 MPa) intensities induced by polyethylene glycol 6000. Our results showed that water deficit did not cause oxidative stress in the seedlings. The most significant effect of water deficit was the depletion of phenolic compounds (flavonoids, catechins, and proanthocyanidins) in seedling roots during the stress imposition period, despite no changes in the phenolic content of the needles. In this context, the lignin content in plant roots was maintained at a level comparable to that of the control. These findings indicate that root lignification in Scots pine seedlings is a stress-resistant process that is important for protecting roots under drought conditions.
Soil contamination by heavy metals (HMs) has intensified with industrialization, mining, and intensive agriculture, creating an urgent need for sustainable remediation strategies. Conventional chemical and physical techniques are costly, disruptive, and difficult to apply at the field scale, emphasizing eco-friendly biological alternatives. This study investigated the combined remediation potential of the microalga Haematococcus pluvialis (H. pluvialis) and three Festuca arundinacea varieties (Nilüfer, Grande II, and Jaguar 4G) for removing cadmium (Cd), lead (Pb), and zinc (Zn) from contaminated soil. Increasing H. pluvialis doses enhanced Cd, Pb, and Zn accumulation in shoots and roots while decreasing Pb bioaccumulation factors. Translocation factors and overall phytoremediation efficiency improved for all metals following microalgal application, with Grande II showing the highest recovery. Post-harvest soil analyses revealed reductions of 57.14%, 20.31%, and 25.46% in Cd, Pb, and Zn concentrations, respectively, alongside a 2.69% decline in soil pH and a 5.34% rise in organic matter. The most effective treatment was 1.5 g kg-1H. pluvialis with Grande II. These findings demonstrate that optimizing microalgal dosage improves metal removal efficiency and supports soil restoration, providing a foundation for sustainable phytoremediation applications.
Non-coding RNAs (ncRNAs) have emerged as central regulators of how plants perceive, integrate, and respond to environmental and biological challenges. Once regarded as transcriptional noise, diverse ncRNA classes, including microRNAs (miRNAs), small interfering RNAs (siRNAs), long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), enhancer RNAs (eRNAs), and piwi-interacting RNA (piRNAs), are now recognized as powerful modulators of gene regulatory networks that shape stress signalling, developmental plasticity, and immune competence. Acting across epigenetic, transcriptional, and post-transcriptional levels, ncRNAs orchestrate chromatin remodelling, RNA stability, redox homeostasis, and hormone signalling to balance growth and defense. This review synthesizes evidence that ncRNA-mediated regulation is highly convergent: identical ncRNA modules are repeatedly recruited across abiotic and biotic stresses and funnel diverse signals through shared regulatory nodes involving hormone networks, reactive oxygen species, and master transcriptional hubs. We highlighted how redox-ncRNA feedback loops, chromatin-embedded ncRNA activity, and cross-layer interactions among miRNAs, siRNAs, lncRNAs, and circRNAs generate synergistic control over stress responses. This review further discusses how ncRNAs encode stress memory and priming, enabling plants to respond more efficiently to recurrent challenges, while also imposing regulatory trade-offs that constrain growth, development, and yield. Despite rapid discovery, ncRNA research remains limited by incomplete mechanistic validation, uncertain epigenetic stability, weak integration with systems biology, and unresolved biosafety and regulatory issues. Addressing these barriers will be essential for translating ncRNA biology into predictive and deployable strategies. By leveraging their capacity to unify signalling and adaptive regulation, ncRNAs represent promising molecular entry points for developing resilient, climate-ready crops.
Wheat (Triticum aestivum L.) is a key global crop threatened by freezing stress, which limits growth and productivity. Cyclophilins regulate critical plant processes, but their role in freezing tolerance remains unclear. In this study, functional assays were conducted exclusively in Arabidopsis thaliana to evaluate cold tolerance. Controlled freezing stress simulations (-5°C) revealed improved stress tolerance traits in transgenic overexpression (OE) plants. TaCYP2-OE lines had significantly improved survival rates (56-60%) in comparison to wild-type line (WT, 33%) after 2 days of stress. Physiological analyses revealed that TaCYP2-OE lines had reduced membrane damage (32-33% lower relative electrolyte leakage and 32-38% lower malondialdehyde content) and elevated proline accumulation (27-31% higher), whereas the superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and lactate dehydrogenase (LDH) activities were significantly higher in TaCYP2-OE lines compared to WT under 2 days stress. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) showed that the expression of antioxidant genes, AtSOD, AtPOD, AtCAT, and ascorbate peroxidase (AtAPX), as well as the stress regulatory genes dehydration-responsive element-binding protein 1a (AtDREB1a) and late embryogenesis abundant protein (AtLEA) of TaCYP2-OE lines were significantly higher than those of WT at 1 day stress. Therefore, TaCYP2 enhances freezing tolerance via membrane protection, osmotic balance, and reactive oxygen species (ROS) detoxification. These findings confirm that TaCYP2 positively regulates freezing stress tolerance and contributes to enhanced cold tolerance in Arabidopsis, with implications for wheat improvement.
Mediterranean basin shrublands are biodiversity hotspots that are currently at risk due to concurrent global change drivers. Here, we investigated the simultaneous effects of a prolonged drought period and increased atmospheric nitrogen (N) deposition on photosynthetic and photoprotective pigments in rosemary (Salvia rosmarinus Spenn.), a widespread iconic Mediterranean woody species, in central Spain. We examined the concentrations and degree of coordination (coupling) of eight photosynthetic and photoprotective pigments under four N addition levels (0, 10, 20, and 50 kg N ha-1 year-1), sampled once immediately after an extended natural summer drought event and again after a two-month recovery period during which plants received ambient precipitation. Our results showed that prolonged summer drought had a pronounced impact, leading to a reduction in chlorophyll, lutein, neoxanthin, and β-carotene concentrations, while increasing the concentrations of photoprotective pigments associated with the xanthophyll cycle, particularly antheraxanthin and zeaxanthin. Drought also caused a disorganisation (decoupling) of pigments, indicating poor metabolic homeostasis under extreme climatic conditions. On the other hand, recovery from drought resulted in increased coordination (coupling). The effects of N addition were minor and mostly associated with greater β-carotene concentrations after drought that we interpreted as indicative of greater oxidative stress. Our findings suggest that extreme drought events will likely have greater effects on pigment metabolism in Mediterranean woody plants than increased N deposition, although interactions between the two global change drivers could amplify physiological disruptions. Our results are among the first to show the metabolic decoupling of plants to concurrent global change drivers, paving the way for new studies focusing on understanding the coordination of physiological responses under stress rather than solely focusing on physiological responses per se.
The TCP transcription-factor family, unique to the plant kingdom, is instrumental in the regulation of plant growth and development. Within this family, the CYC/TB1 clade has been shown to be particularly important in the tillering process of grasses. However, the evolutionary trajectory and functional divergence of CYC/TB1 remain unexplored in Setaria. Here, eight high-quality grass genomes were surveyed and 223 TCP genes were retrieved, allocating 105, 78, and 40 members to the PCF, CIN, and CYC/TB1 subfamilies, respectively. Many TCP genes, including those of the CYC/TB1 class, have undergone purifying selection during evolution, indicating a high degree of functional conservation. Within S. viridis, three CYC/TB1 genes are identified, and they were expressed tissue specifically, with Sevir.9G122200 expressing highly in tiller buds. Heterologous overexpression of Sevir.9G122200 in rice markedly suppresses early axillary shoot formation, demonstrating its conserved repressor activity. This study establishes the first evolutionary framework for the CYC/TB1 clade in Setaria and identifies it as a negative regulator of tillering, providing targets for functional genomics and breeding in related cereals.
Tomato spotted wilt virus (TSWV) is a formidable plant pathogen, inflicting severe economic losses in agriculture due to its broad host range and insect vector transmission. In this study, TSWV was identified in tomato plants in Muş province, Turkey, using molecular methods, and its coat protein (CP) gene was sequenced. The Muş 49 strain shared a remarkable 97% nucleotide similarity with global TSWV isolates and exhibited close phylogenetic relationships with strains from Turkey, Hungary, Bulgaria, and Serbia. Notably, the findings suggest that TSWV genetic diversity is independent of host plant species and geographic location. Beyond genetic characterization, the study explored the antiviral potential of natural stilbene compounds using in silico molecular docking. Remarkably, six of the 13 tested stilbenes exhibited stronger binding to TSWV CP than resveratrol, a well-known antioxidant. Among them, viniferin demonstrated the highest binding affinity (-8.6 kcal/mol), highlighting its promising antiviral potential. These findings suggest that stilbenes may effectively target conserved viral regions and serve as natural inhibitors against plant viruses. Future in vitro and in vivo research will be crucial to validating these promising antiviral candidates. This study not only uncovers new insights into TSWV genetic diversity in Turkey but also paves the way for harnessing natural compounds as innovative plant virus management strategies.
Fusarium solani strain K (FsK) and arbuscular mycorrhizal fungi (AMF) are soilborne symbionts that colonize plant roots and modulate stress responses. While most studies focus on individual microbial partners, understanding multipartite microbial interactions under realistic conditions is essential for designing effective inoculants. Here, we investigated the individual and combined effects of FsK, Funneliformis mosseae (F. mosseae) and Rhizophagus irregularis (R. irregularis) on tomato (Solanum lycopersicum) performance under drought and salinity stress in a greenhouse experimental set up. Under stress conditions, each endophyte showed enhanced root colonization. Co-inoculation with multiple microbes diminished this effect, however the functional outcomes were not directly dependent on the extent of microbial establishment. Under drought, FsK consistently promoted shoot growth, water retention and abscisic acid accumulation, while AMF improved nutrient status. Co-inoculation with FsK and F. mosseae led to synergistic improvements in physiological traits, but only under drought conditions. In contrast, salinity responses were less consistent and revealed functional divergence among microbial partners. These findings demonstrate that context-specific microbial combinations can enhance stress resilience in tomato.
Blue carbon, or carbon fixation, can reduce global CO2 emissions through green ecosystems. The capacity of mangroves to fix atmospheric CO2 is five times higher than tropical or terrestrial land plants. Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo) is one of the most important enzymes for improving photosynthetic efficiency, but a range of sugar phosphates can restrict its activity. The AAA+ protein, RuBisCo activase (RCA), releases this inhibitory sugar-phosphate bound in the active site of RuBisCo by ATP hydrolysis. The present study focuses on understanding the mechanism by which RuBisCo activase regulates RuBisCo in mangroves. In terrestrial plants, RCA supports RuBisCo activity under stress; however, its efficiency diminishes under prolonged or extreme conditions, thereby limiting CO2 fixation. Mangroves, adapted to salinity, may harbour more stress-resilient mechanisms that help maintain photosynthesis. In silico analysis also revealed that mangrove RCA may exist in a hexameric form, with both the α- and β-isoforms indicating a level of structural diversification. Here, we describe a comparative study of RCA isoforms between terrestrial plants and mangroves, highlighting their structural and functional variations in response to environmental stress. We also investigated whether RuBisCo and its molecular chaperone, RCA, contribute significantly to CO2 sequestration in mangroves, or if their roles are minimal or even functionally divergent due to the prevalence of alternative carbon metabolic pathways in these stress-resilient environments.
β-glucosidase, a key enzyme in plant sugar metabolism, belongs to the glycoside hydrolase family and plays a crucial role in plant defense against abiotic stresses. We conducted a comprehensive genome-wide analysis of the watermelon BGLC gene family, identifying eight ClBGLC genes. The molecular weights of these ClBGLCs ranged from 58.17 to 79.53 kDa, and were classified into three subfamilies. Members within the same subfamily exhibited similar gene structures, with these genes being located across five watermelon chromosomes. Collinearity analysis revealed five homologous gene pairs between watermelon and melon, and two with Arabidopsis. Promoter analysis identified several cis-regulatory elements related to plant growth, hormone responses, and abiotic stress responses in the ClBGLC genes. Expression pattern analysis demonstrated that ClBGLCs were expressed across various watermelon tissues. Furthermore, ClBGLC expression was upregulated under salt, drought, and low-temperature stresses, although expression levels varied markedly among genes in response to different stressors. These findings suggest that BGLC genes play a pivotal regulatory role in plant abiotic stress responses. This study provides a foundational basis for future research on the BGLC gene family's functional roles and response mechanisms under abiotic stress conditions.
This study aimed to evaluate the effect of arbuscular mycorrhizal fungi (AMF) on the growth, nutrient uptake, and productivity of chickpea (Cicer arietinum). We investigated the diversity of indigenous AMF in their natural habitat and their effect on the plant and elemental characteristics of chickpea by analysing soil physicochemical properties, root colonisation, AMF spore diversity, and elemental composition of chickpea rhizosphere in two locations (Bhakkar and Khushab, Pakistan). Nitrogen levels of 5.47 g/kg and 4.51 g/kg were found in the rhizosphere soils of Bhakkar and Khushab, respectively. Root colonisation was higher (48.5%) in Khushab (Bhakkar, 35.5%), influencing phosphorus absorption in both regions. Molecular analysis identified 21 AMF taxa, with Glomus and Acaulospora being the most dominant genera. Variations in spore sizes were found, with Glomus measuring 10-191 μm, Acaulospora 125-152 μm, Sclerocystis 110-174 μm, and Gigaspora 65-184 μm. Plant analysis revealed that plant materials from Bhakkar had 1.72% ash, 1.16% fat, 3.78% fibre, and 13.05% protein; samples from Khushab had 1.90% ash, 1.25% fat, 3.24% fibre, and 11.5% protein. Elemental concentrations of chickpea plants from Bhakkar were N = 2.68%, P = 32.98 mg/kg, and K = 33.32 mg/kg, whereas those from Khushab were N = 1.94%, P = 1.17 mg/kg, and K = 43.06 mg/kg. Molecular analysis revealed AMF species with a range of 250-1100 bp. Root colonisation was inversely related to soil phosphorus levels but had a positive effect on plant moisture, fats, and carbohydrates. Morphological and molecular identification showed a relatively high AMF taxa in the rhizosphere of chickpea in both regions. Despite their benefits, the potential of AMF as biofertilisers has not been fully utilised due to prevailing agronomic practices.