Botany - Master Dissertation

Permanent URI for this collectionhttps://kr.cup.edu.in/handle/32116/29

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Investigation of combined genotoxic effect of pesticides on cancer cell lines
(Central University of Punjab, 2013) Anju; Bhardwaj, Pankaj
The use of pesticides in agriculture sector is increasing widely to alleviate crop loss due to pests. To combat problem of pest, various types of pesticides are widely used, out of which insecticide are majorly used. In Malwa region of Punjab lots of insecticides are used. These pesticides when present together they may result in combined response. This combined response can be of three types such as independent, dose addition and interaction (synergism or antagonism). This combined response is very big trouble, such as sometimes even low dose of pesticide which is not toxic individually can result in toxic response in the presence of another pesticide. In the present study, Malathion and Monocrotophos (widely used organophosphorous insecticides) selected for investigation of combined effect. Out of selected pesticide Malathion also detected in water, in present study. Various parameters for genotoxicity evaluation was carried out with exposure of pesticides on HCT-116 WT/ p53 mutant (colon cancer) cell lines. Results of present study suggested that combination of pesticides resulted in more cell death, oxidative stress, membrane instability and imbalanced antioxidative system. It is also evaluated that low dose exposure of pesticides individually and in combination resulted in increased cell survival, may be due to activation of defense mechanism with prior treatment. All parameters evaluated was with short term exposure of 1-2 days, so evaluated long term exposure of ten days and found that this resulted in very less cell number and changed cell morphology. It is observed that both pesticides showed genotoxicity individually as well as in combination. The combined exposure of both pesticides deviate from there expected effect for dose addition towards more toxicity. So it is concluded that there may be some interaction between these pesticides, which may be synergistic in nature, because observed more toxicity.
Assessment of antioxidant potential of phytochemicals in human glioblastoma (U-87 MG) cells
(Central University of Punjab, 2014) Kaur, Manpreet; Mantha, Anil K.
Imbalance between production of reactive oxygen/nitrogen species (ROS/RNS) leads to oxidative stress and has been well documented for mitochondrial dysfunction, a prime cause towards pathogenesis of neurological diseases and cancer. Glioblastoma Multiforme (GBM) is a highly aggressive, invasive and primary brain tumor which shows resistance to chemotherapy and radiotherapy. Superoxide dismutase (SOD) is an antioxidant enzyme that scavenges the production of superoxide radicals and dismutases into H?O? which is further converted into H?O and O? by catalase (CAT) enzyme. Apurinic/Apyrimidinic endonuclease (APE1) is a central enzyme of base excision repair (BER) pathway with two important functions; DNA repair and redox regulation of transcription factors (TFs) responsible for cell survival. In this study, it was seen that oxidative stress induced by endogenously found oxidants H?O? and glucose oxidase (GO) enhanced the activities of both CuZn-SOD and MnSOD in U-87 MG cells. In addition, CuZn-SOD levels were found to be increased in H?O?-induced oxidative stress and MnSOD levels were found to be increased in both H?O? and GO- induced oxidative stress. Further, pretreatment with phytochemicals Curcumin and Quercetin modulated the activities and expression of both forms of SOD studied. The BER-pathway enzyme, APE1 level was found to be decreased in mitochondria of oxidative stress induced U-87 MG cells by H?O? and GO, and in contrast APE1 level was found to be increased in cytosol, which indicates that oxidative stress affects the expression level and sub-cellular localization of APE1. Taken together, these results indicate that in GBM it is more likely that activated SOD a key player of antioxidant system and APE1 a key player in BER-pathway might be facilitating cancer cells to survive in oxidative stress environment.
Evaluation of chemical composition and groundwater and anticancer properties of citrullus colocynthis
(Central University of Punjab, 2014) Kaur, Jatinder; Bhardwaj, Pankaj
Colocynth (Citrullus colocynthis (L.)) is medicinal plant of family Cucurbitaceae which is the native of Turkey. This plant has been used as anti-inflammatory agent, for the treatment of jaundice, skin infections, and urinary diseases from ancient times. It is anti-diabetic, anti-microbial, anti-parasitic, antifungal and anti-oxidant in nature and thought to possess anti-cancer properties also. There is no proper study available regarding its anticancer nature. The aim of the study was to estimate the chemical composition of C. colocynthis fruit and the effect of different extracts prepared from C. colocynthis pulp and seeds on various cancer cell lines. These extracts were found to contain number of secondary metabolites which may further have useful and healing properties. When cancer cells were treated with different prepared extracts, the cell viability of various cancer cells was reduced effectively. Effect of p53 gene was also analysed by using intact p53 as well as p53 mutant cell lines. Antioxidant activity of C. colocynthis was also determined by performing H2DCFDA. Assessment of various antioxidant enzymes such as Catalase, SOD, and Glutathione reductase was also done and found to be present in this fruit. We also analysed the protective effect of C. colocynthis by first treating the cells with pesticides and then with colocynthis extracts. Results indicate that C. colocynthis neutralized the harmful effect of pesticides. The results obtained are encouraging and confirm the value of the use of C. colocynthis as an anticancer agent for fighting against various cancers. It may be helpful to find the exact component responsible for the protective action of C. colocynthis.
Evaluation of genotoxic potential of gropundwater from industrial and farmering sites on colon cancer cell lines
(Central University of Punjab, 2014) Kaur, Rimplejeet; Kumar, Sanjeev
Water is critical component on earth which is essential for life. There are number of reservoirs of water like surface water, groundwater etc. used for drinking, domestic as well as industrial purposes. Numerous studies have shown that water is contaminated by many pollutants like chemicals including pesticides, fertilizers, and heavy metals including arsenic, uranium, and exposure to this contaminated water may cause number of health problems including cancer. It has been found that in Punjab, specifically Malwa region's groundwater is highly contaminated by pesticides and heavy metals which may correlate to several health problems in this region. In this context, mechanisms of genotoxic potential activity of the groundwater, from thermal plant and villages Deon and Joganand, was estimated in the present study. The groundwater caused reactive oxygen species production, mitochondrial membrane depolarization, which ultimately caused cell death. Groundwater also affected the antioxidant system enzymes, so all these factors may be the cause of health problems in this region.
Impact of mitochondrial transplantation on cancer cells
(Central University of Punjab, 2014) Aggarwal, Alza; Bhardwaj, Pankaj
Mitochondria, the powerhouse of the cell, are small granular or filamentous bodies associated significantly with cellular respiration and are the main sources of energy, due to which they are present in maximum number in the organs that require large amounts of energy for doing their function like muscle cells, neural cells, etc. In case of any dysfunction of mitochondria, these organs are most affected culminating in a number of serious multi organs diseases, irrespective of age such as neurogenic weakness with ataxia and retinitis pigmentosa (NARP), or Leigh syndrome (LS), Cancer, etc. Although mutations in mitochondrial genes are common in cancer cells, they do not inactivate mitochondrial energy metabolism, but rather alter the mitochondrial bioenergetics and biosynthetic state. Literature survey also revealed that owing to mitochondrial dysfunction the clinical trial of many anticancer drugs has failed in patients. This study is focused on the impact of mitochondrial transplantation on cancer cells and their drug sensitivity against four human cancer cell lines HCT116 (WT & P53mutated), HepG2 and MCF7. The normal cell's Mitochondria was transplanted into cancer cells and then evaluated the Impact of transplantation of mitochondria from healthy cells into cancer cell upon their growth, ROS production and their drug sensitivity. The results of this study revealed that the healthy mitochondria transplanted to cancer cells decrease carcinogenesis and have drug sensitivity. So, it may be used as futuristic cancer remedy.
Pollen induced stress in human lung carcinoma A549 cells
(Central University of Punjab, 2014) Atri, Rekha; Dhiman, Monisha
The pollen grains are important aeroallergens causing allergic disorders like rhinitis and asthma. Chronic asthma often leads to carcinogenesis in lungs. The various mediators of inflammation like reactive oxygen and nitrogen species (ROS/RNS), cytokines and chemokines form a link between oxidative stress, inflammation and cancer. This study involves induction of oxidative stress by pollen extracts derived from two varieties of prickly poppy in small lung carcinoma A549 cells. To rule out the pathway responsible for pro-inflammatory and oxidative stress enhancer we used the inhibitors of the classical ROS sources such as NADPH oxidase, Xanthine oxidase, mitochondria and general ROS scavenger (NAC). In addition, we also used NF-kB blocking peptide to see whether blocking NF-kB pathway can attenuate the pollen induced stress. It was observed that there was no significant difference in inflammation and/or oxidative stress in the purified pollen antigen versus the crude pollen extract. Lipid peroxidation was significantly increased in pollen extract treated cells whereas no change in MPO activity was observed. The NF-kB levels were significantly reduced in pollen extract treated cells whereas the levels were normalised when treated with NOX inhibitor (DPI) and mitochondria uncoupler (FCCP) suggesting the role of mitochondrial and phagocytic ROS in activating NF-kB. This study signifies the role of pollen as aero-allergens which are very common in Malwa region of Punjab as a key player in enhancing the allergy related complications and chronic lung diseases.
Status of endogenous antioxidants defense in preconditioned contrasting chickpea in precondioned contrasting chickpea (cicer arietinum) seedlingls
(Central University of Punjab, 2014) Jairath, Ankur; Kumar, Sanjeev
Plants experience various abiotic stresses of low or elevated temperature, drought and heavy metals. Chickpea (CicerarietinumL.) is one of the important crop belongs to the family Leguminosae and it is the second most important grain legume plant in the world after common bean. Chickpea has evolved as a spring crop in warmer climate of West Asia. Therefore, it is prone to chilling injury as in north India; chickpea is cultivated in winter season. To protect the plant from chilling injury, preconditioning the plants with mild drought stress might triggers the signalling pathways that will prepare the plants to combat lethal cold stress. This is determined by monitoring the various cell responses at different phases. Therefore antioxidative enzyme activity of the SOD, APX, GPOX, LOX, CAT enzymes was done. In our study, both the varieties, PDG3 and GPF2 showed equalvariation towards SOD, CAT, GPOX, and APX activity. Enhanced SOD, CAT & GPOX activity in both the varieties showed increased tolerance towards cold stress. Less LOX activity showed preconditioning decreased membrane damage in preconditioned seedlings. Thus, preconditioning with mild drought stress has enhanced tolerance capacity of the plant during the cold stress.
Charaterization of G-SSR markers in rhododendron arbreum
(Central University of Punjab, 2014) Kaur, Harmeen; Bhardwaj, Pankaj
Himalayan region is one of the important hotspot, having Rhododendron arboreum as major species. Anthropogenic activities and unsustainable use of the plant for its fruits, flower and wood by local people has raised questions about the sustainability of the species. In order to answer these questions, it is important to first know the genetic structure of the population. This study is focused on the characterization of the G-SSR markers for Rhododendrons and to study the population genetic structure. A set of 38 primers out of 41 are characterized on three population of Rhododendron arboreum, a total of 30 individuals. Allele number at different loci came to be in range of 2 to 14 with an average of 5.2 alleles per locus. Level of heterozygosity observed was high which varies from 0.422 to 0.917 (He) and 0.167 to 0.933 (Ho) with a polymorphic information content 0.104 to 0.811. Out of 38, about 19 loci deviated from the Hardy Weinberg Equilibrium. Genetic diversity was high in these populations with an average gene flow of 5.436. Based on the information from the loci, there was no observed linkage disequilibrium and bottleneck effect. Software DARwin and STRUCTURE, irrespective of geographical location, revels the presence of two original populations with admixed genetic stock. These studied loci will help in the further genetic mapping of the genes and genotyping studies in Rhododendron arboreum.
Development of genomic microsatellite markers in commiphora wightii
(Central University of Punjab, 2014) Kaur, Gaganpreet; Bhardwaj, Pankaj
Commiphora wightii is an important medicinal plant, growing in arid to semi- arid conditions. It is widely distributed in tropical regions of Africa, Madagascar and Asia. In India, the specie is found in South- Western India and parts of Central India. The plant on incision secretes an oleo gum resin, which is used to treat various ailments since ancient times. The oleo gum resins of plant contain guggulsterones which have great medicinal value. The other uses of gum are in perfumery, calicoprinting, dyeing silk and cotton, fumigation and incense for which the plant is getting exploited by unfair tapping methods. This plant has been listed in red data book so conservation of this plant is the need of present. For this it is necessary to know about the genetic diversity and population structure of the plant. Microsatellite markers are markers of choice for such studied because they are reproducible, co- dominant and show high levels of polymorphism. In the present study genomic SSR markers were developed. A total of 22 primer pairs were designed for which, 1913 clones were analysed for fragments containing microsatellite regions and 338 clones were found positive and are selected for sequencing. Sequencing results show a total of 212 fragments with perfect repeats, interrupted repeats and compound repeats (> 5 repeats). These primer pairs can be used in genetic diversity analysis of naturally growing populations of Commiphora wightii. They can also be used in MAS, genome selection during gene introgression in plant breeding, genome mapping and gene tagging. This will help in planning the conservation strategies as well as rational utilization of this endangered plant.
Influence of preconditoning on water relations and pigment system during reproductive stage in winter sown chickpea.
(Central University of Punjab, 2016) Jarial, Poonam; Thakur, Sanjeev K.
Chickpea is self –pollinated legume crop which was believed firstly originated in South-Eastern Turkey and parts of Syria. Chickpea is the second most important pulse crop in the world. In Northern India, chickpea is grown in winter season. Preconditioning the plant with mild drought stress may trigger the various signaling pathways which will prepare the plants to conquer lethal cold stress. In this study, PBG1, PBG5 and GPF2 variety are more sensitive to the chilling stress, which were preconditioned with mild drought stress and then exposed to lethal cold stress. To see the effect of preconditioning various cell responses were monitored by measuring ELI, Relative Water Content and chlorophyll content at different stages of chickpea. All genotypes generated a variable response. Overall, relative water content and chlorophyll content were significantly higher in preconditioned PBG1, GPF2 and PBG5, which are sensitive to chilling stress. The study showed increase tolerance capacity in preconditioned plant towards chilling temperature and improve tolerance against chickpea
Analyzing Reproductive Capabilities Of Chickpea In Cold Environment
(Central University of Punjab, 2018) Selpair, Shweta; Kumar,Sanjeev
Chickpea is winter sown legume crop. In northern India, crop faces chilling stress during the month of January. Chilling stress causes detrimental effects on the reproductive stages of the crop which leads to the abortion of flowers, pod setting and seed yield. Preconditioning is a process in which plants are treated with mild drought stress to induce plant defense system against chilling stress. Effect of preconditioning was studied on five genotypes PBG1, GPF2, PDG3, PDG4 and PBG5. In this experiment, performance was evaluated on the basis of their seed yield. Reproductive structures were studied with the help of compound, scanning electron and confocal microscopy. Results have shown that PBG5 has performed best while PBG1 worst on the basis of seed yield among the five genotypes.
Investigating The Germinating Behaviour Of Chickpea (Cicer Arietinum L.)
(Central University of Punjab, 2018) Uniyal, Ajay Prakash; Kumar, Vinay
Seed germination is the crucial stage in plant development process. It commences with water imbibition and culminates with radicle protrusion. This process has been observed to be occurred in sequential three phases, phase I with rapid water uptake followed by phase II (Plateau Phase) and phase III in which radicle extrusion takes place. Water imbibition during phase I triggers the sets of events that causes some physiological and biochemical changes in seeds as starch degradation by activation of hydrolytic enzymes etc that are strongly related to plumule and radicle growth. The present investigation aims to investigate the germinating behaviour of Chickpea (Cicer arientium L.) and biochemical alteration during the three phases of germination. Phase I is found to be from 0-20 hours, Phase II from 20-32 hours and Phase III after 32 hours. During germination, both starch and proteins were found to be degraded to provide the developing embryo with sugars and free amino acids respectively and the content of reducing sugars and total sugars were found to be increased. The result indicates that the reserve seed storage mobilisation may be helpful to proceed the seedling growth.
Isolation And Culturing Of Photobionts From Antarctic Lichen
(Central University of Punjab, 2018) Acharya, Sayanya; Bast, Felix
Lichen is the symbiotic association in between algal and fungal partner. Lichens having various economic importance. The main objective is to isolate and culture of the photobionts from the lichen samples collected from Antarctica. Trebouxia sp. is extremely psychrophilic and fastidious in growth, found in the lichen samples. Both samples belong to genus Umbilicaria, Umbilicaria antarctica and Umbilicaria aprina. In the lichen samples Gyrophoric acid is found as secondary metabolites. First to isolate the photobionts, we use micromethod and for the culturing use 3M BBM media for 45-60 days at 22-25o C temperature. Then subculturing process is done and followed by DNA extraction. The extracted DNA was kept for further use in sequencing purpose. The isolated and cultured photobionts can be further studied using DNA sequencing, SEM, and confocal microscopy to get a clear idea about the photobionts and its internal organelles; these will help in proper identification of the photobionts as well as the lichen. Further phylogenetic studies of photobionts and coevolutionary studies between photobionts and mycobionts which may be done by extensive taxa sampling and through use of molecular systematics techniques.
Characterization And Culture Of Snow Alga Chlamydomonas Nivalis From Antarctica
(Central University of Punjab, 2018) Samal, Archana; Bast,Felix
The snow alga Chlamydomonas nivalis is the reason of colouring the snow orange-red. For current study sample was collected from Dalk Glaciers, Larsmann hills, Eastern Antarctica. This is the most studied alga of snow-fields but cytology, morphology, and taxonomy are still unsolved. Our experiment focuses on comparative growth rate study of algal culture at two different temperature one is of 4°C and other is of 20-22°C and its morphology as well as life cycle study. Experiment confirmed that algal culture at 20-22°C was growing at a higher rate than the culture at 4°C. Identification and characterization were done on the basis of light microscope and phase microspore studies. Phase contrast microscope picture signifies the presence of different types of algal cells in culture. Flow cytometry strongly support the phase contrast microscope data and signifies the presence of algal cells in different stages of its life cycle.
Differential Alternative Splicing Analysis Under Abiotic Stress Conditions In Barley (Hordeum Vulgare)
(Central University of Punjab, 2018) Bisht, Diksha; Bhardwaj, Pankaj
Biotic and abiotic stress factors adversely affect the growth and development of plants, living in constantly changing environment. Plant abiotic stress responses seem to be frequently associated with the process of alternative splicing. Alternative splicing generates multiple products from a single gene by variable splicing of exons. RNA-seq utilizes NGS platforms to produce massive amount of sequence data. Using transcriptomic data, we tried to identify the differentially spliced genes in barley under heat and cadmium stress. Raw RNA seq data was pre-processed and aligned with reference barley genome. Then differential alternative splicing was analyzed on the basis of intron excision. In this study, a total of 196 significant differentially spliced clusters were found under two abiotic stress conditions, among them 47 and 149 clusters were associated with cadmium and heat stress condition, respectively. Overall, 133 fully annotated clusters and 63 unannotated clusters were observed. Further, annotation was performed for the significant genes to determine their functions. Some of these significant genes encode for splicing factors, stress tolerance protein, non snRNPs and antioxidants. From this study, it was concluded that barley undergo differential spliced events to maintain its homeostasis under different abiotic stress conditions.
Development And Characterization Of SSR Markers In Calotropis Procera
(Central University of Punjab, 2018) Vandana; Bhardwaj, Pankaj
Calotropis procera is a weed growing around the arid and semi-arid areas of the tropical and subtropical regions. The species has the potential to replace cotton and resolve the high rates of cotton-based products and at the same time reduce the use of synthetic non-biodegradable fibres. It has been used in various medicinal formulations from a long time. There is a need for developing molecular markers to facilitate breeding and genetic improvement of its varieties. Out of 12.6 million raw reads, 84636 reads were obtained after quality check and filtering. 93.7% of alignment rate and 76.31% of core genes were detected in Bowtie and BUSCO assessment, respectively. An average of 5.5 SSRs/Mb was obtained in 72.349 Mb transcriptome. 25 SSR markers were selected for characterization on 30 individuals representing 3 populations. 13 SSR loci were found polymorphic. In the population structure analysis a moderate level of genetic diversity (Na=3.9231, He=0.633) was predicted. The genetic variation among the population was found to be only 7% whereas within the population, it was 93%. The mean Shannon?s information index obtained was 1.1, showing significant diversity richness in the populations. The Dendrogram showed that the population from Bathinda and Ambala are closely related to each other than those from Barnala. The gene flow between Bathinda and Ambala population was found significantly high (6.439). The present set of loci had a high PIC value of 0.506 that makes them very informative to be further used for large scale studies.
Identification Of Ganoderic Acids And Derivatives From Fruiting Bodies Of Ganoderma Lucidum
(Central University of Punjab, 2018) Pradhan, Agrah; Kumar,Sanjeev
Ganoderic acids and other bioactive triterpene metabolites from crude extract of wood decaying fungus Ganoderma lucidum from two different hosts Acacia and Azadirachta were to be isolated as proposed. The method for extraction was simple and divided into three parts, 1. Solvent selection, 2. Dissolving metabolites into solvent for maximization of number of detected metabolites, 3. Extraction of metabolites from solvent. A variety of methods for maximizing dissolution in solvent based on pharmaceutical extraction protocols were employed. Sonication was chosen as the best method with detection of 5 distinct spots (metabolites) from TLC. Ethanol based sample extracts were packed inside silica gel column while using chloroform as the mobile phases in column chromatography. Some metabolites were isolated and confirmed by TLC. Due to time bound project work the further isolation was paused and the crude extracts were forwarded for GC-MS detection and analysis of constituent metabolites. After preparation of sample and analysis in gas chromatography mass spectroscopy (GC-MS), 16 metabolites were found in chloroform extract of ganoderma having azadirachta as host and 24 metabolites were found from acacia host associated fungal crude extract. The detected metabolites were alkaloids, phenols, fatty acids (and precursors), esters, di-ene compounds and sterols (terpenes).Terpenes (C28-Phytosterols) were found with molecular weight 396-398. 7,22-Ergostadienol with area 2.91% and 1.19% is found in Acacia and Azadirachta samples. Moreover another metabolite 7,22- Ergostadienone with area 0.61% is also found in Acacia (Host) sample this metabolite has various bio-active properties like anti-inflamatory and cytotoxic properties. These are derivatives of the ergosterols found in fungal body. These structures are products of lanosterol, (squalene derived products) they have many anthropocentric exploitive uses and they resemble the metabolite backbone (derivatives) we were initially looking for.
Comparative Analysis of Metabolites in Contrasting Chickpea Cultivars
(Central University of Punjab, 2018) Ghosh, Alokesh; Kumar, Vinay
Phenolics are the major class of plant secondary metabolites. Among half of the plant phenolics are the flavonoids. The production of these metabolites induces during stresses and thus helps the plant to cope with the environment. Anthocyanins, a class of flavonoids protects plant from harmful UV rays, attracts the pollinators and seed dispersal, helps in mimicry and also helps in the root nodulation process in legumes. Chickpea is a leguminous and major pulse crop plant, shown in the winter season faces lots of stress. Anthocyanins and other phenolics help the plant to tolerate such stresses. Desi cultivars have more phenolics and anthocyanin as compared to the kabuli one. Flowers have higher concentration of anthocyanin than in leaves. Untargeted metabolomics profiling of the dry powdered mature seed samples of contrasting chickpea cultivars detects primary and secondary metabolites such as fatty acids, phenolics, phelols, terpenes, esters,sugar,etc in the seeds of chickpea.
To Study Callus Induction from Explants of Nicotiana tabacum
(Central University of Punjab, 2018) Reetu; Kumar,Vinay
Due to a great importance of callus most of the work has been carried out on regeneration aspect of callus including somatic embryogenesis, which is very important aspect of plant tissue culture and crop improvement. A large number of plants have been regenerated in plant tissue facility using various explants in phytohormone supplemented MS media. In present proposal, three selected phytohormone (NAA, BAP and 2,4-D) in different concentration combinations have been used for callus induction using leaf and seedling as explants in tobacco. It was found that best combination for induction of callus is BAP and NAA as compared to BAP and 2,4-D. Color, texture and size of callus were assessed after 25 days of culture. Rate of growth was found more in the culture media containing BAP and NAA combination. It was observed that color of callus was greenish in culture media having BAP and NAA and yellowish brown in the culture medium containing BAP and 2,4-D. In conclusion, this work help to establish that both seedling and leaf explants can be used for callus formation with appropriate supply of phytohormones.
Amelioration of Salinity stress in Chickpea (Cicer arietinum L.) variety PBG5 through foliar spray of Ascorbic acid
(Central University of Punjab, 2018) Kumar, Rajneesh; Choudhary, Krishna
Chickpea (C. arietinum L.) is the second most important legume crop sown as a winter crop in northern parts of India, and its productivity is constraint by several abiotic stress, and sat stress is one of the important factor for significant yield loss. Spraying the plants with ascorbic acid might trigger the signaling pathways that will prepare the plants to combat lethal salt stress. In this study, chickpea variety PBG5 was sprayed with ascorbic acid along with 50 mM of NaCl stress and further analyzed for various parameters. To see the effects of ascorbic acid spray various cell responses were monitored by measuring membrane damage (ELI & MDA content), water status (RLWC), Chlorophyll content and Antioxidant enzyme activity (SOD and CAT) along with growth and total biomass. Present study clearly reflected that foliar spray of ascorbic acid enhances the tolerance capacity of the plants and prevents noticeable damage in them. To conclude whole study, treatment with ascorbic acid has ability to improve the tolerance against salt stress in chickpea.