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ThesisItem Open Access Biochemical defense response and genetic basis of resistance against Ascochyta blight (Ascochyta rabiei (Pass.) Labr.) in chickpea (Cicer arietinum L.)(Punjab Agricultural University, 2024) Abassy, Omer; Dr Upasana RaniChickpea (Cicer arietinum L.), the second-largest global pulse crop, is under threat from highly variable nature of Ascochyta rabiei, causing widespread disease damage under favourable environmental conditions. This disease, Ascochyta blight is a major concern worldwide, with insufficient resistant sources and frequent breakdowns due to rapid pathogen evolution. This study evaluated 238 kabuli chickpea genotypes for resistance to Ascochyta blight (AB) over the growing seasons from 2019-2022 at Punjab Agricultural University under artificial epiphytotic conditions, identifying 18 resistant lines (GLK 10-40, GLK 20054, GLK 20055, FLIP 10-298C-IFC-S2, FLIP 10298C-IFC-S1, CS-3-E-24, FLIP 09 256C-55, FLIP 07-314C-57, FLIP 10-243C, FLIP 09-194C, FLIP 08-104C, FLIP 04-219C, ICCV 55233, ICCV 55215, ICCV 55135, ICCV 55108 and ICCV 155141) belonging to different genetic backgrounds with disease scores of 1.0 to 3.0, whereas seventy-three (73) lines demonstrated a moderate level of disease resistance with a disease rating of 3.1-5.0. The activities of defense-related enzymes (PAL, TAL, PPO, POD) associated with phenol metabolism along with lignin and total phenol content were compared among six kabuli chickpea lines, viz., five resistant (GLK 10-40, GLK 20055, FLIP-09-194C, FLIP-04-219C, ICCV 55215) and one susceptible (GLK 17301) treated lines that exhibited differential responses to Ascochyta blight at tested time intervals of 48, 96, 144 and 240 hr post-inoculation respectively. The perusal of data showed increased activity of all enzymes (PAL, TAL, PPO, POD) till 96 hr after inoculation, whereas the non-enzymatic contents such as lignin and total phenols showed the maximum enzyme activity till 144 hr after inoculation. However, enzyme and non-enzymatic activity remained almost constant with no significant difference in all control/untreated resistant and susceptible line(s) at different time intervals. For understanding the genetic basis of resistance, the F2 population comprising of 162 plants derived from the cross, GLK 24096 (resistant) and L 556 (susceptible) was assessed under artificially epiphytotic conditions against local isolate of Ascochyta rabiei. Of 162 plants evaluated, 39 plants were grouped in resistant (R ) category (disease score ≤ 5.0 ) and 123 were classified as susceptible (S) with score of five and above. Consequently, the F2 plants exhibited a segregation ratio of 1R:3S, suggesting that a monogenic recessive gene governed resistance to Ascochyta blight. Of thirty four SSR markers associated with Quantitative Trait Loci (QTLs)/genes were used for confirmation of resistance in identified eighteen resistant and six susceptible lines, only 19 SSR markers depicted polymorphism. Notably, SSR markers, STMS11, TA194, TS12, TA146, and TA2 exhibiting significant polymorphism, indicated tight linkage to Ascochyta blight resistance. Phylogenetic analysis showcased distinct clustering based on resistance, offering insights into the genetic basis of resistance. These findings informed about targeted resistance breeding and sustainable disease management with defence-related enzymes/identified markers holding promise for developing superior, resistant and resilient chickpea varieties contributing to crop sustainability and productivity.ThesisItem Embargo Characterization of mycotoxic Fusarium species associated with ear rot of maize(Punjab Agricultural University, 2024) Harinder Singh; Dr. Harleen KaurEar rot samples of maize (Zea mays L.) were collected during Kharif and spring seasons from different districts of Punjab. In all, 48 Fusarium isolates were obtained and purified for further studies. These isolates were grouped into six major groups based on morphological characteristics viz., colony color, pigmentation, type of mycelium, pattern of growth, growth rate and sporulation. Two species of Fusarium viz., F. verticillioides and F. proliferatum were identified using ITS, TEF-1α, RPB2, VERT- 1/VERT-2 gene specific primers. The sequences of four representative isolates-Fus 15, Fus 28, Fus 44 and Fus 45 when submitted to BLAST showed 99-100 per cent similarity with F. verticillioides and Fus 48 isolate had shown 97.91 per cent similarity with F. proliferatum. Further, out of 47 isolates of F. verticillioides VERTIF-1/VERTIF-2 primers 40 isolates with fumonisin producing ability. FUM13F /13R primers identified 36 isolates of F. verticillioides with fumonisin producing ability. FUM1 primers revealed that out of 48 Fusarium isolates, 41 isolates possessed FUM1 gene. Genetic diversity analysis of Fusarium isolates using 20 SSR primers revealed high degree of polymorphism and clustered 48 isolates into two major groups and seven subgroups. The primer 3H07 amplified maximum number of loci (6) ranging from 100-1400 bp in size and had highest PIC value of 0.76. On comparing the clusters of morphological, molecular and pathological characterization defined correlation could not be obtained among morphological characteristics, SSR pattern and pathological behavior of Fusarium isolates. For standardization of inoculation technique, we used four techniques viz., needle inoculation into the cob, toothpick inoculation in the ear, needle inoculation into the silk and spray silk inoculation, out of which needle inoculation into the cob technique gave highest mean disease index of 26.44 and 36.77 per cent during spring and Kharif seasons, respectively. Higher disease severity of FER was obtained when cobs were inoculated at 7 days after silking in both spring (19.34 %) and Kharif (32.16%) seasons. On comparing the seasons, Kharif season showed comparatively more disease severity (25.25 %) than spring season (17.1 %). Using this standardized technique, forty-five advanced maize germplasm lines were screened artificially against FER during Kharif 2021, disease severity varied from 27.77 to 96.29 per cent being lowest (27.77 %) in LM13×Teosinte-1-2-1-1-1 (BC1F1) and highest (96.29 %) in LM14×Teosinte-1-1-1-1 (BC1F1) line. Out of 45 lines, none of lines showed resistance reaction to FER, however 13 lines were found moderately resistant. Pathological behavior of all Fusarium isolates was studied on four hybrids (PMH 1, PMH 2, PMH 10 and PMH 13) and two inbreds (LM 13 and EML 173) of maize during spring 2022. Based upon their virulence, heatmap was generated and all isolates were clustered into seven groups. Group I isolates comprising Fus 15 (Ludhiana), Fus 23 (Jalandhar) and Fus 48 (Sirmour) were highly aggressive giving maximum disease severity of 68.74 per cent. Among the hybrids, PMH 1 showed minimum mean disease severity (34.16%) giving moderately resistant reaction to all Fusarium isolates. Among the inbreds, LM 13 showed minimum mean disease severity of 41.16 per cent giving moderately resistant reaction. The fumonisin production potential of F. verticillioides isolate- Fus 15 and F. proliferatum isolate-Fus 48 was observed on PMH 1 and PMH 2 hybrids under artificial inoculation conditions. Concentration of Fumonisin B1 (FB1) produced by Fus 15 and Fus 48 isolates was comparatively higher in PMH 2 than PMH 1 hybrid. Maximum fumonisin production of both FB1 and FB2 in maize grains was estimated at 21 days after incubation.ThesisItem Embargo Resistance gene identification in Vigna species infected with Begomovirus(Punjab Agricultural University, 2024) Kuppuraj J; Sirari, AsmitaThe successful intra and inter-specific hybridization in Vigna revealed the dominant nature of yellow mosaic disease resistance with both monogenic and digenic gene action with dominant epistasis in Vigna mungo and digenic gene action with complementary epistasis in Vigna radiata. Further, differential expression of different classes of resistance genes in both compatible and incompatible Vigna – mungbean yellow mosaic India virus interactions was investigated. The qPCR primers were designed for amplifying plant resistant protein domain - specific regions (NB-ARC; SgS3; RdRp3) from genome of different Vigna species. Resistance protein domain specific sequences were shortlisted (with candidate genes, type species) and retrieved from NCBI database, followed by multiple sequence alignment through MEGA11 software on different Vigna species genome. A total of 37 qPCR primers with three domain - specific regions were designed using Primer3Web version 4.1.0 online software, were amplifying with specific amplicon size (90 to195 bp) and moreover without primer dimer formation. Most of the primers were amplified on genomic DNA of four resistant and two susceptible genotypes of different Vigna speices genome. Total RNA was isolated and cDNA panel was prepared from mock and MYMIV inoculated leaf tissues of both resistant and susceptible genotypes of different Vigna species at 1, 5, 10, 15, 20, 25 and 30 days post inoculations. SYBR green chemistry was performed in RT-qPCR to study differential gene expression in both compatible and incompatible interactions. All the three resistant protein domains i.e. NB-ARC, SgS3 and RDR were found to be involved in YMD resistance in Vigna radiata and Vigna umbellata with absence of NB-ARC functional, only in Vigna mungo. On the other hand, expression of mungbean yellow mosaic India virus specific coat protein gene AV1 was higher in susceptible Vigna radiata genotype (SML1082) as compared to Vigna mungo and vice-versa in case of replication gene AC1. Low level of expression of both the AV1 and AC1 genes was also observed in resistant genotypes of Vigna with first experimental evidence of Vigna umbellata as host of mungbean yellow mosaic India virus.ThesisItem Embargo Understanding sunflower - Macrophomina phaseolina interaction under water stress conditions(Punjab Agricultural University, 2023) Navkiran Kaur; Sharma,PankajA total of 35 isolates of Macrophomina phaseolina causing charcoal rot were collected from eleven kharif and rabi crops from different districts of Punjab and adjoining states. Macrophomina phaseolina isolated from sunflower field, Ludhiana was characterized on cultural and morphological and molecular basis. The isolates infecting sunflower showed highest sequence homology of ITS genomic region with M. phaseolina isolate from Indore, Madhya Pradesh NCBI accession no. MT127404. This isolate was further used to standardize the inoculation techniques under field and polyhouse conditions. Tooth pick inoculation technique was found most effective for large scale screening while sorghum grain inoculation method was proficient under polyhouse conditions. A set of 71 sunflower genotypes were evaluated using tooth pick inoculation technique against charcoal rot under irrigated and restricted irrigation conditions. Four genotypes viz; 75B, EC6078261, OPH137, and OPH 172 were found moderately resistant under irrigated conditions whereas only two genotypes 75B and EC6078261 showed moderately resistant reaction in restricted irrigated conditions. The present study was first attempt to investigate the water stress-Macrophomina phaseolina interaction on different components of physiological, plant growth and yield parameters along with disease incidence. A significant reduction in total chlorophyll content (54.78 %), relative water content (47.06 %), membrane stability (51.75 %), sugar (69.00 %), protein (13.90 %) and proline (56.66 %) content was observed with inoculum density of 4 g/kg soil at soil moisture levels of 40 % and 60 %. The inoculum density of 4 g/kg of dry soil resulted in maximum mortality during seedling stage and growth period. Thus, varied inoculum density and water stress conditions made the sunflower plants more prone to dismissive physiological alterations. It led to biomass reduction viz; root length (41.27 %), root width (76.74 %), root weight (85.37 %), shoot length (56.71 %) and shoot weight (79.06 %). Thus it resulted in per plant seed yield reduction (95.45 %) at 52.62 to 100 % disease severity at higher inoculum density (4g, 6g, and 8g/kg dry soil). The expression levels of the defense related genes LOX, ACCO1, PAL, SOD, and APX were also modified under varying degree of water stress and inoculum density. The strong correlation between low soil moisture and disease severity implies that the proper irrigation of the field can be suggested as cultural management practices for the farmers.ThesisItem Restricted Management Of Sclerotinia Stem Rot Of Rapeseed-Mustard Incited By Sclerotinia Sclerotiorum (Lib.) De Bary(Punjab Agricultural University, 2023) Yadav, Monika; Sandhu, P.S.Sclerotinia stem rot caused by Sclerotinia sclerotiorum, is an important fungal disease of rapeseed and mustard crop because of its destructive nature. In the present study, 112 genotypes of rapeseed-mustard screened against stem rot disease. Ten genotypes (EC223400, EC765482, IC268306, IC296703, IC491169, IC491178, IC491197, IC491290, IC531377 and IC538699) were found moderately resistant to the pathogen. Early sowing of brassica during first week of October results in lower incidence of the disease and higher yield compare to the late sowings. Poison food technique was used to evaluate botanicals and fungicides under in vitro condition. Among the botanicals maximum per cent inhibition in mycelial growth was observed in case of Chenopodium album at 20 per cent concentration. Whereas, in case of fungicides complete inhibition (100%) was observed in combination fungicides picoxystrobin 7.05%+ propiconazole 11.7% w/w SC, azoxystrobin18.2% w/w + difenoconazole 11.4% w/w SC and fluopyrum 17.7% w/w + tebuconazole 17.7% w/w SC at a concentration of 5, 10 and 25 ppm, respectively. Under in vitro conditions, combination fungicides were more effective against S. sclerotiorum compared to contact fungicide mancozeb 75 % WP which resulted in lower inhibition of pathogen growth even at higher concentrations. The same set of botanicals and fungicides were used under in vivo conditions as a foliar spray application. Chenopodium album at 40 % concentration was observed most effective in reducing the per cent lesion length as compared to the control. Combination fungicide picoxystrobin7.05%+ propiconazole 11.7% w/w SC resulted in lower lesion length development and highest yield as compare to the control in Brassica juncea var. RLC 3.ThesisItem Restricted Virulence pattern of leaf and neck blast isolates (Pyricularia oryzae Cavara) from basmati rice and identification of resistance donors(Punjab Agricultural University, 2023) Anjali; Jain, JyotiThe blast disease caused by Pyricularia oryzae was prevalent throughout the Punjab state with leaf blast incidence ranging from 9.47 to 54.26 percent and neck blast incidence ranging from 23.73 to 46.11 percent respectively. The highest disease incidence and disease severity were observed in the agro-ecological Zone-III comprising of Bathinda, Sangrur, Barnala and Muktsar districts on variety Pusa Basmati 1401 followed by Pusa Basmati 1509 and Pusa Basmati 1121. Leaf and neck blast isolates were found to be variable with respect to colony colour, elevation, margin, form, sectoring, zonation, mean radial growth (mm), spore size and extent of sporulation. Three leaf blast isolates (LB-3, LB-10 and LB-11) and five neck blast isolates (NB-51, NB-52, NB-65, NB-83 and NB-89) were categorizedas fast growers with Kr value of more than 8.1mm/24hr. All the tested isolates sporulated better on Oatmeal agar medium than on Paspalum grass medium. Genetic analysis of the leaf and neck blast isolates using SSR markers revealed high degree of polymorphism with PIC values ranging from 0.44 to 0.90 and 0.44 to 0.93 respectively discerning the isolates into two major clusters. Studies on virulence pattern and cross-infectivity exhibited substantial variability among leaf and neck blast isolates on a set of eight basmati varieties. Univariate and multivariate analysis of isolates originating from leaf or neck portions of basmati rice with respect to various components of aggressiveness viz., infection efficiency (IE), Incubation period (IP50), Latent period (LP), Lesion size (LS), Sporulation intensity (SI), Disease incidence (DI) and Disease severity (DS) demonstrated their cross-pathogenic behaviour. However, the neck blast isolates proved to be more aggressive on rice leaves than leaf blast isolates on rice necks. Among the different components of aggressiveness, IE, LS and SI were positively correlated with each other, whereas IP50 and LP50 showed a negative correlation with all the aggressiveness components. Out of 96 germplasm lines screened against leaf and neck blast phases separately under artificial inoculation conditions, none of the genotypes was found to be resistant to both phases. Only six genotypes namely, IR 67417-75-3-2-2-3, IR 72860-62-1-1-3, Acharamati, Basmati C122, Pusa Basmati 1637 and Tetep showed moderately resistant reaction to both leaf and neck blast phases with susceptibility index <3. Ten genotypes IR 67017-73-1-4, IR 67418-131-2-3-3-3, IR 68726-3-3-1-2, IR 68740-3-2, IR 70418-112-1-2, Basmati Surkh 161, JJ-92, Tulsi Phoola, UPR-3519-18-1-1 and IET 15392 exhibited moderately resistant reaction to leaf blast but were moderately susceptible to neck blast. Under natural epiphytotic conditions at hotspot location (RWRC Malan, H.P.), twelve genotypes namely IR 67017-73-1-4, IR 67417-75-3-2-2-3, IR 68726-3-3-1-2, IR 68740-3-2, IR 70418-112-1-2, IR 71740-7-3-1, Basmati C122, JJ-92, Tulsi Phoola, IET 15392, Pusa Basmati 1637 and Tetep showed resistant reaction to leaf blast and moderately resistant reaction to neck blast. The present study established that resistance to the leaf blast phase does not necessarily imply resistance to the neck blast, indicating a need to evaluate rice genotypes for both the leaf and neck blast phases in the ongoing blast breeding programs. Furthermore, the new insights gained into the biology of P. oryzae, its organ specificity, comparative aggressiveness, and the genetic sources of resistance will be useful in designing better management strategies.ThesisItem Restricted Sheath rot and its association with discolouration of rice(Punjab Agricultural University, Ludhiana, 2021) Sarbjit Kaur; Anju BalaThe present investigation was undertaken with the objective to find out the association of pathogens causing grain discolouration and sheath rot of rice and to identify the resistance sources against sheath rot in rice germplasm. Four pathogens viz., Sarocladium oryzae, Fusarium verticillioides, Fusarium fujikuroi and Albifimbria terrestris were found associated with sheath rot and grain discoloration of rice. The fungal pathogens were inoculated on plants using syringe inoculation, grain insertion method and spray inoculation on 2 varieties, PR 114 and PR 121. The variety PR 114 was more susceptible to fungal disease as compared to PR 121. Plants inoculated with grain insertion method showed high disease incidence followed by syringe inoculation method. Among all the pathogens, S. oryzae caused maximum per cent disease incidence and severity followed by F. verticillioides, F. fujikuroi and A. terrestris. The highest disease progress was also observed in plants inoculated with S. oryzae. All the pathogens produced similar disease symptoms in rice. All the pathogens caused chaffiness of panicles, significant reduction in seed germination, seedling length and vigor index was observed in highly infected seeds. The seed quality deterioration was directly proportional to the intensity of seed infection. Further, the effect of fungal metabolites extracted from discoloured grains was observed on physiological and biochemical parameters by dipping seeds in these metabolites for different durations. The seed quality losses were increased with the increase in duration of seed dipping in fungal metabolites. Significant reduction in total sugars and total proteins along with an increase in defense related enzymes, amino acids, proline, total phenols and malondialdehyde content was recorded. Fifty advanced breeding lines of promising rice germplasm were screened against sheath rot by inoculating three pathogens S. oryzae, F. verticillioides and F. fujikuroi using two inoculation techniques (syringe inoculation and grain insertion). Among all the lines evaluated against sheath rot, RYT 3886 was found to be resistant and RYT 3836 was moderately resistant to all the three pathogens.ThesisItem Embargo Biodiversity of seed-borne pathogenic mycoflora of wheat (Triticum aestivum L.) in Punjab(Punjab Agricultural University, 2023) Rubleen Kaur; Tak, Parminder SinghA total 105 grain markets of 23 districts of Punjab were surveyed for study of seed-borne mycoflora during 2021-22 and 654 grain samples were collected. Grains were visually inspected for incidence of Karnal bunt (KB), Black point (BP) and shrivelling. Prevalence of Karnal bunt, Black point and shrivelled grains was 10.24%, 55.20% and 79.51 per cent, respectively. Grain samples from Patiala and Mansa were found to have statistically lowest KB, black point and shrivelled seeds while highest incidence was observed in Pathankot followed by Hoshiarpur and Mohali. Three pathogenic and nine opportunistic fungi were isolated from these samples. Fungal diversity in Sarhali Kalan samples (66.67%) was highest, followed by Sirhind-1, Jalandhar, Rajpura (58.3% each). Bikhi, Nabha, Dadehra, Bucho mandi, Panjgrain (16.67%) carried low mycoflora load. All the samples collected met the minimum seed certification standards for germination. Maximum vigour index I and II of 3327.6 and 26.46 was observed in grain samples collected from Doraha-1 and Shahkot, respectively. Prevalence of Alternaria spp. (94%) was highest followed by Fusarium spp. (64%), Aspergillus flavus (48%) whereas, low prevalence was observed for Exserohilum sp. and Leohumicola sp. (2% each). The fungal isolates were characterized on the basis of morphology and cultural characteristics into 11 fungal genera belonging to 7 families which included Alternaria alternata, Bipolaris sorokiniana, Curvularia verrucosa, Curvularia spicifera and Exserohilum rostratum from Pleosporaceae family (34%); Aspergillus flavus, A. japonicus and Penicillium digitatum, from Aspergillaceae (17%); Fusarium equiseti from Nectriaceae (8%); Epicoccum purpurascens from Didymellaceae (8%); Leohumicola sp. from Incertae sedis (8%), Chaetomium globosum and Pseudothielavia terricola from Chaetomiaceae (17%) and Rhizopus stolonifer from Rhizopodiaceae (8%). Sequencing of Curvularia verrucosa, Bipolaris sorokiniana, Curvularia spicifera, Aspergillus japonicus, Pseudothielavia terricola, Alternaria alternata and Fusarium equiseti was done and cultures have been deposited in MTCC repository, Chandigarh. Thirty nine fungal isolates were inoculated on three wheat varieties and determined for seed health, foliar blights and carryover studies. Opportunistic fungi such as Curvularia sp. (isolate CUS-1), Aspergillus sp. (isolates AF-1, AF-7), Rhizopus sp. (RIZO-1), Penicillium sp. were found to reduce germination under field conditions. Highest average foliar blight severity was observed in Gurdaspur (26.98%) as compared to 20.60 per cent in Ludhiana. Bipolaris isolate BIS-1, Aspergillus flavus AF-1, Curvularia CUT-1, Alternaria ALT-6 and ALT-10 resulted high severity in Ludhiana while Fusarium FUS-1, Rhizopus RIZO-1, Drechslera DRH-3, Aspergillus flavus AF-7 and Alternaria ALT-8 were highly pathogenic in Gurdaspur. The seed inoculated with these fungi induced foliar blight, affected the germination as well as severity of foliar blight however, they could not be re-isolated in all the cases indicating their primary role only up to seedling stage. Overall, Alternaria spp. had the highest frequency in grain market samples as well as carryover studies.ThesisItem Unknown Detection, variability and translocation of Potato virus Y (PVY) infecting potato in Punjab and identification of host plant resistance(Punjab Agricultural University, 2023) Belludi, Rakesh; Sharma, AbhishekThis study aimed to investigate the variability of Potato virus Y (PVY) infecting potato crop of Rabi season in Punjab, encompassing symptomatology, serology and molecular aspects. The study revealed prevalent symptoms, such as mild mosaic, chlorotic patches, interveinal chlorosis, puckering, veinal necrosis and stem necrosis, on potato plants caused by various potato viruses including PVY. ELISA tests confirmed the presence of six out of seven viruses in potato samples. Among them, PVX exhibited the highest incidence (78.08%), followed by PVYN (51.37%), PVM (20.55%), PVYO/C and PVS (13.01%). PLRV exhibited a lower incidence (2.74%) and no samples tested positive for PVA. Notable regional variations noticed, with Ludhiana showing high incidences of PVYN (78.43%), PVYO/C (31.37%), PVM (33.3%) and PVS (35.29%). Conversely, Kapurthala displayed a 100% PVX incidence, while Jalandhar had the highest PLRV (19.05%) incidence. In nucleic acid-based detection through RT-PCR, the primer NIb2F/3R effectively amplified the desired ~350bp amplicon in all PVYN and PVYO/C seropositive samples. Whereas, the qPVY NIb F2/CP R2 primer generated the ~1.5kb fragment in only 24 out of 67 samples, further confirmed via BLASTn sequence analysis as PVYO, PVYNTN and PVYN-Wi strains. Further virus translocation studies revealed a wide range of carryover rates, spanning from 40% to 92.46%, from infected mother plants to daughter tubers, exhibiting significant variation in viral titre across genotypes and among tubers. Spatial translocation within the tuber's root zone revealed that the bottommost tuber displayed a relatively lower PVY titre. Interestingly, freshly harvested tubers exhibited a higher viral titre at the heel end (stem end) compared to the rose end eyes in all examined tubers. After storage, a contrasting trend emerged in sprouted tubers, where the highest PVY titre was found in rose end eyes and the lowest in heel end eyes. In germplasm screening against resistance to PVY, one germplasm (KP -16-19-14) displayed high resistance (HR), while 16 exhibited resistant (R), 20 moderately resistant (MR), 22 moderately susceptible (MS), 10 susceptible (S) and two were highly susceptible (HS) to PVY. Following PVY inoculation, significant reduction was observed in chlorophyll fluorescence parameters, viz., Fv/Fm, Y(II), qP, and qL values in susceptible cultivars, accompanied by notable increases in NPQ, Y(NO) and qN values. Susceptible genotypes displayed yield losses ranging from 13.64 to 58.84% with viral titre ranging from 0.014 mc to 419.673 mc. Finally, a farmer-friendly and cost-effective vertical flow assay (VFA) was developed for rapid PVY detection in potato leaf samples. The sensitivity of the developed assay was equivalent to commercially utilized ELISA. Specificity test confirmed assays precision for prevalent PVY strains and its non-specificity to other potato viruses. Developed assay exhibited 95% accuracy compared to RT-PCR.