Defence mechanism through a biochemical pathway in Kabuli chickpea genotypes against Ascochyta blight

Defence mechanism through a biochemical pathway in Kabuli chickpea genotypes against Ascochyta blight | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Defence mechanism through a biochemical pathway in Kabuli chickpea genotypes against Ascochyta blight Omer Abassy, Satvir Grewal Kaur, Upasana Rani This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3886689/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Chickpea ( Cicer arietinum L.), the second-largest global pulse crop, plays a crucial role in providing essential minerals and dietary fiber. Ascochyta blight, caused by the necrotrophic pathogen Ascochyta rabie i, poses a substantial threat to chickpea cultivation. Contemporary cultivars often lose resistance to this disease, necessitating improved management strategies. In this study, six Kabuli chickpea genotypes underwent at treated and controlled conditions, identifying GLK 10–40, GLK 20055, FLIP 09-194C, FLIP 04-219C, and ICCV 55215 resistant and GLK 17301 susceptible to Ascochyta blight. The exploration into the biochemical responses of these genotypes revealed dynamic shifts in enzymatic activities and biochemical components post-fungal infection. PAL enzyme activity witnessed a remarkable 45.5-fold increase at 96 hours post inoculation in the resistant genotype GLK 10–40, underscoring its essential role in the early defense cascade against Ascochyta blight. TAL and PPO activity peaked at 96 hours post inoculation notably in GLK 20055 and GLK 10–40 respectively, emphasizing its participation in the initial defense response. POD activity, a crucial element in plant immunity, reached its peak at 96 hours post inoculation, particularly in the resistant line GLK 20055, signifying prolonged defense mechanisms. Lignin content exhibited a consistent increase till 144 hour post inoculation notably in GLK 20055, highlighting its structural contribution to defense against Ascochyta blight. Total phenol content, crucial in resisting microbial infection, showed heightened levels till 144 hour post inoculation in resistant line FLIP 04-219C maintaining sustained high levels. These findings unveil the biochemical intricacies of chickpea defense mechanisms against Ascochyta blight, laying the groundwork for targeted breeding or genetic engineering. The identified genotypes, such as GLK 10–40, GLK 20055, and FLIP 04-219C, hold promise for developing resilient chickpea cultivars to counter this challenging disease, crucial for ensuring global food security. Chickpea Kabuli biochemical enzymes resistant defense Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 1. Introduction Chickpea ( Cicer arietinum L.) holds significance as the second-largest pulse crop, cultivated and consumed worldwide, predominantly in Afro-Asian nations. Global chickpea production reached approximately 15.87 million tons with an area of 16.01 million hectare in 2022. In India, chickpea cultivation covers an area of approximately 10.94 million hectares resulting in an annual production of 11.91 million tons (FAOSTAT 2022 ). Chickpeas constitute a valuable reservoir of crucial minerals such as zinc, iron, calcium, copper, potassium, and phosphorus, in addition to containing vitamin B and dietary fiber. This elevated dietary fiber content is especially beneficial for those with insulin sensitivity or diabetes (Pittaway et al., 2008 ). Chickpea genotypes can be categorized into two main types: desi and kabuli. This classification is primarily determined by seed size, shape, and color. Desi chickpeas are predominantly cultivated in Asia and Africa, whereas kabuli chickpeas are commonly found in the Mediterranean region and are extensively grown in North America, particularly in Mexico and the United States (Chandra-Hioe et al 2016 ). While chickpea production has seen recent growth, it still struggles to meet the demands of an increasing vegetarian population. The challenge lies in enhancing production and productivity, given the limitations in expanding cultivable areas. Factors like human intervention, evolving cultural practices, and climate change contribute to an increase in diseases, affecting crop yields. A total of 172 chickpea pathogens have been reported globally, including approximately 67 fungi, 3 bacteria, 22 viruses, and 80 nematodes, posing significant biotic stresses on chickpea cultivation (Nene et al., 1996 ). However, only a handful of these pathogens can cause widespread damage, leading to epidemics that significantly impact global chickpea production. Diseases like Ascochyta blight, Botrytis gray mold, Fusarium wilt, dry root rot, black root rot, collar rot, Phytophthora, Pythium root rot, seed rot, and seedling rot pose a major economic threat. Others have sporadic occurrences and localized distributions (Johansen et al., 1994 ). Aschochyta blight by the highly virulent necrotrophic pathogen Ascochyta rabiei (Pass.) Labr. (syn. Didymella rabiei ) in chickpeas (Owati et al., 2017 ). This pathogen can infect crops at various stages, leading to significant yield reduction, particularly during flowering and podding stages (Wise et al., 2011 ). The disease is more prevalent under cool and wet conditions, with temperatures ranging from 15 to 25°C and relative humidity between 65% and 100%, favoring its development and spread. Severe infections result in reduced seed quality and complete yield loss (Pande et al., 2005 ). Aschochyta blight manifests as lesions on foliar and stem parts, leading to stem breakage and seed rot. Symptomatic features include bended or lengthened darker brown-red lines on leaflets, concentric circles of pycnidia on leaves, stems, and pods, and 3–4 cm length brown lesions with black spots on stems and petioles (Li et al., 2015 ). Modern cultivars frequently lose resistance and become susceptible to diseases over time. The use of fungicides for disease management is often impractical and economically unfeasible (Reddy et al., 1990). The mechanisms underlying resistance during the host-pathogen interaction in both resistant and susceptible genotypes remain poorly understood. Plants, in response to pathogens, possess effective defense mechanisms where static and dynamic secondary metabolites play crucial roles as local or systemic resistance factors (Redman et al., 1999 ). The interaction between the host and pathogen induces signalling molecules in the plant system, leading to the production of antimicrobial phenolic substances. These substances participate in various biochemical processes. In certain diseases, infection is marked by an increased synthesis of phenolic compound precursors and oxidation products of phenolics, such as quinones, which demonstrate heightened toxicity to microorganisms (Bahr et al 2016 ). Understanding biochemical defense mechanisms is crucial for defining resistance in specific genotypes. Induction of resistance in plants involves physiological and biochemical changes, including callose, phytoalexins, lignin, and plant pathogenesis-related proteins. Enzymes such as peroxidase (POD), phenylalanine ammonia-lyase (PAL), and polyphenol oxidase (PPO) play a vital role in the defense activities of plants against various abiotic and biotic inducers, contributing to resistance. In chickpeas, higher activity of PPO, PAL, and peroxidase, along with low catalase activity in leaf samples, appears to be a significant biochemical factor contributing to resistance against Ascochyta blight infections (Angelini et al., 1993 ). The understanding of these processes is essential for developing strategies to enhance plant resistance against diseases. Thus, this study significantly contributes to enhancing strategies for the effective management of Ascochyta blight. Through the identification of novel resistant sources in Kabuli chickpea and a deeper understanding of the biochemical defense mechanisms involved, our findings provide valuable insights that can be applied in developing robust and sustainable approaches to combat this detrimental disease. 2. Material and methods 2.1 Plant material and pathogen A total of 238 different Kabuli chickpea germplasm lines were screened against Ascochyta blight ( Ascochyta rabiei ) under artificial epiphytotic conditions during the rabi seasons of 2019-20, 2020-21 and 2021-22 at the Research Farms of the Pulses Section, Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana. Out of the 238 lines, six lines namely, GLK 10–40, GLK 20055, FLIP 09-194C, FLIP 04-219C, ICCV 55215, and GLK 17301were chosen based on exhibiting both resistant and susceptible reactions (Table 1 ). Two sets, comprising five resistant and one susceptible line each, were sown. In one set, the lines were inoculated with Ascochyta rabiei local isolate using the foliar spray method, while the other set remained uninoculated (control). Artificial raining effect and relative humidity were maintained with perfo-spray system for disease development during the daytime from 10.00 am to 04:00 pm regularly for 21 days. Sampling was done at intervals of 48, 96, 144, and 244 hours after inoculation. Collected samples were placed in an icebox and promptly stored at -20°C for subsequent enzymatic analysis, including phenylalanine ammonia lyase (PAL), tyrosine ammonia lyase (TAL), and polyphenol oxidase (PPO), peroxidase (POD). For the estimation of lignin and total phenols, samples were gathered in an icebox and preserved in methanol (80%) for total phenols and ethanol (90%) for lignin, both stored at -20°C. Table 1 Resistant and susceptible kabuli chickpea lines against Aschochyta blight ( Aschochyta rabiei ) for biochemical analysis S.No Line Name Disease Score Reaction Type 1 GLK 10–40 1.33 Resistance 2 GLK 20055 2.17 Resistance 3 FLIP 09-194C 2.83 Resistance 4 FLIP 04-219C 2.50 Resistance 5 ICCV 55215 1.83 Resistance 6 GLK 17301 9.00 Susceptible 2.2 Disease severity scale The data were collected using a rating scale of 1–9 points, as outlined in Table 2 , and further detailed in instances where the susceptibility check revealed maximum disease. The lines were classified as highly resistant (HR), resistant (R), moderately resistant (MR), susceptible (S), or highly susceptible (HS) based on this rating scale (Table 2 ). Table 2 Rating scale (1–9) against Ascochyta blight of chickpea Disease score Description Reaction types 1 No visible symptom Highly Resistant (HR) 2 Small/minute lesions were observed on foliage of plants, mostly not visible Resistant (R) 3 Few scattered lesions were observed on foliage and stems of plants, mostly visible after careful examination 4 Lesions were detected on foliage and stems of few plants but were not damaging. Moderately resistant (MR) 5 Lesions were commonly observed on foliage and stems but damage was less 6 Lesions commonly detected on foliage and stems; 25.0% or one fourth of the plants would be killed. Susceptible(S) 7 Lesions would be commonly observed on foliage and stems, 50.0% or half of the plants would be killed. 8 Lesions were extensively observed on all of the plants, girdling of stem and branches visible, 75.0% or three fourth of the plants would be killed. Highly Susceptible (HS) 9 Lesions were extensively observed on all of the plants tissues, girdling was extensive on all of the plant parts including stems and branches and > 75.0% or more than three fourth of plants would be killed. 2.3 Extraction and estimation of enzymes The enzymatic antioxidants such as phenylalanine ammonia lyase (PAL), tyrosine ammonia lyase (TAL), polyphenol oxidase (PPO), peroxidase (POD) lignin, and total phenols were estimated in twigs of chickpea at four stages such as S1 (48 hours), S2 (96 hours), S3 (144 hours), and S4 (240 hours). To determine enzymatic antioxidants, the enzymes were extracted using extraction buffers at 4°C to minimize enzyme denaturation, and the assays were conducted at 30°C. For extraction of PAL and TAL enzymes, 0.1 g of fresh chickpea plant samples were homogenized in about 2 ml of Tris HCl (0.1 M) buffer having a pH of 7.5 and containing β-mercaptoethanol (5 mM). Subsequently, they were centrifuged for 25 m at temperature of 4ºC and 10,000 rpm. Then, the supernatant was utilized to estimate the PAL and TAL. For estimating the activity of PAL enzyme the reaction mixture containing 2.5 ml of 30 mM phenylalanine prepared in 0.05 M sodium borate buffer (pH 8.8) and 100 µl of enzyme extract was incubated at 37ºC for 1 hr. The reaction was stopped by the addition of 0.3 ml of 5 N HCl. The absorbance was recorded at 290 nm against blank (simple water). No incubation was done in case of control treatment and the reaction was stopped right away. Additionally, 5–40 µg of cinnamic acid was utilized to create a standard curve that was further used to calculate the activity of PAL. Furthermore, the activity was expressed as µg of cinnamic acid formed min − 1 mg − 1 fresh weight, whereas for in case of TAL enzyme the reaction mixture for TAL was also incubated at 37ºC for 1 hour and contained 1 ml of 33 µM tyrosine, 50 µl of enzyme extract and also sodium borate buffer (1.35 ml) having a pH of 8.8. Furthermore, the reaction was stopped by the addition of 5 N HCl (0.1 ml). The absorbance was recorded at 310 nm against the water blank. As there was no incubation in case of control, the reaction was stopped right away. The standard curve made with coumaric acid (5–40 µg) was used to calculate the activity of TAL. Its activity was further expressed as µg of coumaric acid formed min − 1 mg − 1 fresh weight. For extraction and estimation of PPO enzyme activity Freshly weighed chickpea plant samples (0.1g) were crushed in 2 ml of ice-cooled 0.1 M potassium phosphate buffer (pH 6.8) containing 2-mercaptoethanol (5 mM), 1% PVP and 1 mM EDTA before being centrifuged at 10,000 rpm for 20 min. The supernatant collected was used as an enzyme extract. The reaction mixture consisted of 100 mM 4-methyl catechol, buffer and enzyme extract. The change in absorbance was recorded at 410 nm for 3 min at an interval of 30 sec. One unit of enzyme activity was defined as an increase of 0.01 in absorbance min − 1 mg − 1 of fresh weight. For extraction and estimation of POD enzyme activity freshly weighed chickpea plant samples (0.1 g) were homogenized in 2 ml of ice-cold 0.1 M potassium phosphate buffer (pH 7.0) containing 2 β-mercaptoethanol (5 mM), 1% PVP and EDTA. The material was centrifuged at 10,000 rpm for 20 min after being filtered through muslin cloth. An enzyme extract was prepared using the collected supernatant. The supernatant was collected and used as enzyme extract. The reaction mixture consisted of 0.05 M guaiacol, 100 µl enzyme extract and 0.8 M H₂O₂. The change in absorbance was recorded at 470 nm for 3 min at an interval of 30 sec using the UV-visible spectrophotometer. The enzyme activity of peroxidase was expressed as a change in absorbance min − 1 mg 1 of fresh weight. For extraction and estimation of lignin content fresh plant samples (0.1 g) of treated and untreated chickpea plants were homogenized in 2 ml of 95% ethanol. The homogenate was then centrifuged at 10,000 rpm for 20 min. The supernatant was discarded and the pellet was washed thrice with 95% ethanol and twice with freshly prepared ethanol: hexane (1:2) solution. Washed pellets were dried overnight at 45°C. After being dissolved in 25% acetyl bromide made in acetic acid, the dried pellet was incubated at 70°C for 30 min before being cooled at ambient temperature. The extract so obtained was used for estimating the lignin content. The reaction mixture consisted of 100 µl extract, 180 µl of 2 N NaOH, 20 µl of 7.5 M NH₂OH.HC1 and 1.6 ml acetic acid. After centrifuging this mixture for 5 min, the intensity of colour generated was measured at 280 nm. The simultaneous running lignin standard curve (5–40 µg) was used to calculate the lignin content. The, so obtained, lignin content was expressed as mg g − 1 fresh weight. For extraction and estimation of total phenol content fresh chickpea plant samples (300 mg) were homogenized in 3 ml of 80% methanol and refluxed with methanol for 1 hr. The refluxed content was filtered and the pellet was re-extracted with 80% methanol. The filtrate from these two extractions was then mixed and methanol was used to make up the volume to 5 ml. This methanolic extract was used to calculate the amount of total phenols. Methanolic extract (0.5 ml) was taken in test tubes and evaporated to dryness. To this, 6.5 ml distilled water and 0.5 ml Folin's-Phenol reagent (prepared in 1:1 with distilled water) were added and vortexed to mix well. Then 1.0 ml saturated solution of sodium carbonate was added and the mixture was kept at room temperature for one hour. The intensity of blue colour developed was read spectrophotometrically at 760 nm against the reagent blank. The standard curve was prepared using gallic acid (10–50 µg). Total phenols, so obtained, were expressed as mg of gallic acid equivalent 100 g − 1 fresh weight. 3. Result In the current study, six Kabuli chickpea genotypes were cultivated under both controlled and infected conditions. These genotypes were classified according to disease incidence, with GLK 10–40, GLK 20055, FLIP 09-194C, FLIP 04-219C, and ICCV 55215 identified as resistant genotypes, and GLK 17301 designated as a susceptible genotype (refer to Table 1 ). Biochemical parameters were assessed in the leaves of these genotypes at four stages and various hours after inoculation (HAI). 3.1 Biochemical reactions in response to the occurrence of Ascochyta blight 3.1.1 Phenylalanine Ammonia-Lyase (PAL) PAL enzyme activity in response to Ascochyta rabiei inoculation in chickpea genotypes, with results indicating an increase in activity in resistant chickpea lines (GLK 10–40, GLK 20055, FLIP-09-194C, FLIP-04-219C, and ICCV 55215) displayed higher PAL enzyme activity than the susceptible line (GLK 17301), peaking at 96 hrs and declining thereafter in treated lines(Table 3 ). Treated plants of GLK 10–40 displayed highest PAL activity (33.06 µg cinnamic acid formed min − 1 g − 1 FW at 96 hrs and 18.612 µg cinnamic acid formed min − 1 g − 1 FW at 144 hrs) and significant increase of 45.5-fold at 96 hrs followed by 26.40-fold rise at 144 hrs. Conversely, treated GLK 17301(susceptible genotype) consistently showed lower mean PAL activity (2.52 µg cinnamic acid formed min − 1 g − 1 FW) than resistant treated lines with considerable increases compared to control treatments at different tested time intervals (Figs. 1 and 2 ). Table 3 Phenylalanine ammonia-lyase (PAL) activity in kabuli chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with Ascochyta rabiei . Name Phenylalanine Ammonia Lyase (PAL) activity (µg cinnamic acid formed min − 1 g − 1 FW) 48 hrs 96 hrs 144 hrs 240 hrs Treated Control Treated Control Treated Control Treated Control GLK 10–40 6.503 0.722 33.063 0.723 18.612 0.705 0.987 0.705 GLK 20055 4.356 0.465 18.374 0.460 11.215 0.431 0.706 0.529 FLIP-09-194C 7.658 0.542 21.729 0.583 8.202 0.504 0.601 0.462 FLIP-04-219C 3.932 0.384 12.338 0.303 8.708 0.375 0.497 0.365 ICCV 55215 4.872 0.360 12.128 0.340 7.755 0.372 0.511 0.374 GLK 17301 1.218 0.158 6.674 0.188 2.005 0.163 0.225 0.189 C.D. 1.088 2.881 1.627 0.183 SE(m) 0.349 0.925 0.522 0.059 SE(d) 0.494 1.308 0.738 0.083 C.V. 18.828 13.21 13.528 14.118 3.1.2 Tyrosine Ammonia-Lyase TAL is also vital enzyme of defense pathways and results showed increased TAL activity at 96 hrs post-inoculation which started decreasing afterwards. Comparisons revealed significant increase in TAL activity in all treated genotypes. Treated resistant line, GLK 20055 exhibited the highest activity (68.75 µg coumaric acid formed min − 1 g − 1 FW) at 96 hrs while susceptible genotype, GLK 17301 had the lowest (1.68 µg coumaric acid formed min − 1 g − 1 FW) at 240 hrs (Table 4 ). Control lines of both resistant and susceptible genotypes maintained constant TAL activity. FLIP-09-194C (resistant line) had the maximum TAL activity at 48 (6.57-fold increase) and 144 (10.72-fold rise) hrs respectively while GLK 20055 (resistant line) reached the highest value at 96 and 240 hrs with 15.74-fold and 2.49-fold rise respectively. The susceptible treated genotype, GLK 17301 consistently showed lower average TAL activity (4.91 µg coumaric acid formed min − 1 g − 1 FW) as compared to resistant treated lines with slight increases at different time intervals compared to controls. These findings depict dynamic changes in TAL enzyme activity in response to fungal infection in different chickpea genotypes, with the time frame now expressed in hours (Figs. 3 and 4 ). Table 4 Tyrosine ammonia-lyase (TAL) activity activity in kabuli chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with Ascochyta rabiei . Name Tyrosine Ammonia Lyase (TAL) activity (µg Coumaric acid formed min − 1 g − 1 FW) 48 hrs 96 hrs 144 hrs 240 hrs Treated Control Treated Control Treated Control Treated Control GLK 10–40 15.225 6.441 52.250 6.157 22.010 6.794 8.333 6.922 GLK 20055 17.897 4.034 68.750 4.368 29.113 4.475 9.873 3.951 FLIP-09-194C 18.054 2.745 33.216 2.990 29.373 2.740 3.510 2.833 FLIP-04-219C 15.971 3.059 29.667 3.373 22.328 3.676 4.637 3.417 ICCV 55215 15.873 4.377 26.039 4.078 17.456 4.637 6.505 4.265 GLK 17301 3.887 1.691 9.868 1.623 4.216 1.539 1.686 1.544 C.D. 1.423 5.795 3.658 0.895 SE(m) 0.457 1.86 1.174 0.287 SE(d) 0.646 2.63 1.661 0.406 C.V. 6.947 14.364 13.406 8.787 Fig 3. Different range of intervals between the maximum and minimum change in the activity of Tyrosine ammonia-lyase (TAL) in resistant and highly susceptible kabuli chickpea lines in both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs post-inoculation with Ascochyta rabiei . 3.1.3 Polyphenol Oxidase (PPO) Results of PPO enzyme revealed increased activity till 96 hrs after inoculation, reaching the maximum limit and then declining. GLK 10–40 (resistant) showed the highest PPO activity (345.83 unit’s min − 1 g − 1 FW) at 96 hrs while GLK 17301 (susceptible) had the lowest (52.25 unit’s min − 1 g − 1 FW) at 240 hrs in treated genotypes (Table 5 ). Control lines maintained constant PPO activity. PPO activity was highest in resistant treated genotypes i.e. GLK 20055 (233.50 units min − 1 g − 1 FW) at 48 hrs, GLK 10–40 at 96 (345.83 units min − 1 g − 1 FW) and 144 (275.50 units min − 1 g − 1 FW) hrs respectively and ICCV 55215 (93.50 units min − 1 g − 1 FW) on 240 hrs. Comparisons showed significant increases in PPO activity in treated plants with a 5.24-fold rise in GLK 10–40 at 96 hrs. The treated susceptible genotype, GLK 17301 consistently exhibited mean lower PPO activity (119.59 unit’s min − 1 g − 1 FW) with significant increases at different time intervals compared to controls. These findings depict dynamic changes in PPO enzyme activity in response to fungal infection in different chickpea genotypes, with the timeframe expressed in hours (Figs. 5 and 6 ). Table 5 Polyphenol oxidase (PPO) activity activity in kabuli chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with Ascochyta rabiei Name Polyphenol oxidase (PPO) units min − 1 g − 1 FW) 48 hrs 96 hrs 144 hrs 240 hrs Treated Control Treated Control Treated Control Treated Control GLK 10–40 156.250 63.333 345.833 66.000 275.500 64.500 76.250 63.750 GLK 20055 233.500 74.417 311.917 78.167 271.250 76.917 88.500 77.583 FLIP-09-194C 166.667 68.083 289.500 66.250 244.000 61.500 76.500 68.417 FLIP-04-219C 151.167 61.500 237.750 66.417 217.500 63.667 74.167 58.500 ICCV 55215 177.000 67.333 254.833 67.500 215.250 64.250 93.500 65.750 GLK 17301 102.083 47.500 166.500 46.583 157.500 47.833 52.250 47.500 C.D. 14.438 21.664 13.813 8.792 SE(m) 4.634 6.954 4.434 2.822 SE(d) 6.554 9.834 6.27 3.991 C.V. 6.202 5.909 4.316 5.796 3.1.4 Peroxidase (POD) Peroxidase enzyme (POD) prevents accumulation of hydrogen peroxide by catalyzing different substrates, however this study indicated that its activity increased after 96 hrs of inoculation in all treated plants with the highest enzyme activity observed in the resistant plants. The maximum enzymatic activity was observed resistant genotype, GLK 20055 at 96 (247.418 ∆A min − 1 g 1 FW) hrs, followed by 144 ( 197.948 ∆A min − 1 g 1 FW), 48 ( 183.678 ∆A min − 1 g 1 FW) and 240 (97.290 ∆A min − 1 g 1 FW) hrs respectively while GLK 17301 genotype showed the lowest on 240 (49.92 ∆A min − 1 g 1 FW) hrs (Table 6 ). The control plants maintained a consistent level of POD activity. Comparisons between treated and control plants revealed significant increases in POD activity in treated resistant genotype, GLK 20055 showing 8.2-fold increase at 96 hrs, whereas the susceptible treated genotype GLK 17301 exhibited average lower POD activity(62.23 ∆A min − 1 g 1 FW), with significant increases at different time intervals compared to controls. These findings illustrate dynamic changes in POD enzyme activity in response to fungal infection in different chickpea genotypes, with the timeframe expressed in hours (Figs. 7 and 8 ). Table 6 Peroxidase (POD) activity activity in kabuli chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with Ascochyta rabiei. Name Peroxidase activity (POD) (∆A min − 1 g 1 FW) 48 hrs 96 hrs 144 hrs 240 hrs Treated Control Treated Control Treated Control Treated Control GLK 10–40 150.253 58.143 213.600 50.667 194.900 51.650 93.195 51.925 GLK 20055 183.678 39.900 247.418 30.143 197.9483 36.292 97.290 34.075 FLIP-09-194C 150.737 44.682 235.860 40.438 183.200 47.817 96.045 42.587 FLIP-04-219C 181.157 75.900 198.055 74.758 195.360 77.013 96.622 60.960 ICCV 55215 166.675 40.153 209.120 44.050 172.773 47.657 71.925 45.573 GLK 17301 65.705 31.765 67.633 26.458 65.670 30.938 49.923 31.917 C.D. 15.484 9.429 11.405 11.995 SE(m) 4.97 3.026 3.661 3.85 SE(d) 7.029 4.28 5.177 5.445 C.V. 6.917 3.548 4.678 8.239 3.1.5 Lignin content Plant defence component lignin increased till 144 hrs post-inoculation in all treated lines, with higher levels in resistant lines. GLK 20055 (resistant genotype) had significantly maximum lignin content at different time intervals (48, 96, 144 and 240 hrs), while minimum lignin content was observed in GLK 17301(susceptible genotype) (Table 7 ). Comparisons revealed noteworthy increase in lignin content in treated resistant genotype, GLK 20055 with 4.79-fold rise at 144 hrs, whereas susceptible treated genotype, GLK 17301 consistently exhibited lower lignin content compared to controls at different time periods. These findings illustrate dynamic changes in lignin content in response to fungal infection in different chickpea genotypes, with the timeframe expressed in hours (Figs. 9 and 10 ). Table 7 Lignin content in kabuli chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with Ascochyta rabiei. Name Lignin content (mg − 1 g 1 FW) 48 hrs 96 hrs 144 hrs 240 hrs Treated Control Treated Control Treated Control Treated Control GLK 10–40 5.005 3.630 8.787 3.130 11.158 3.105 4.190 3.267 GLK 20055 6.966 3.158 9.585 3.009 14.947 3.174 5.253 3.218 FLIP-09-194C 4.090 2.907 5.651 2.131 8.032 2.537 3.851 2.708 FLIP-04-219C 4.371 2.657 7.188 2.734 8.422 2.328 3.769 2.397 ICCV 55215 5.502 2.615 6.624 2.952 7.150 2.312 4.102 2.766 GLK 17301 2.827 2.135 4.019 2.041 5.796 2.143 2.241 2.025 C.D. 0.834 1.304 0.547 0.581 SE(m) 0.268 0.418 0.176 0.187 SE(d) 0.379 0.592 0.248 0.264 C.V. 10.106 12.126 4.193 7.674 3.1.6 Total phenols Phenols provide structural integrity and support the plants defence mechanism by counteracting/nullifying pathogen attack. Increase in total phenol content was achieved until 144 hrs after inoculation in all treated lines, with higher levels in resistant lines compared to the susceptible line. The zenith of total phenol content in FLIP-04-219C (resistant treated genotype) of 14.310 mg − 1 g 1 FW was achieved at 144 hrs and 7.164 mg − 1 g 1 FW at 240 hrs respectively followed by resistant treated genotypes, GLK 10–40 at 96 (11.899 mg − 1 g 1 FW) hrs and ICCV 55215 at 48 (10.352 mg − 1 g 1 FW) hrs respectively, while susceptible treated genotype, GLK 17301 showed minimum phenol content of 3.023 mg − 1 g 1 FW at 48 hrs, 3.346 mg − 1 g 1 FW at 96 hrs, 4.992 mg − 1 g 1 FW at 144 hrs, and 2.224 mg − 1 g 1 FW at 240 hrs post-inoculation respectively as compared to control lines (Table 8 ). Comparisons revealed significant increases in total phenol content in treated plants, notably a 4.22-fold rise in resistant line, FLIP-04-219C at 144 hrs but susceptible genotype, GLK 17301 exhibited lower average total phenol content (1.40-fold) with substantial increases at different time intervals compared to controls. These findings illustrate the dynamic changes in total phenol content in response to fungal infection in different chickpea genotypes, now expressed in hours (Figs. 11 and 12 ). Table 8 Total phenol content in kabuli chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with Ascochyta rabiei Name Total phenol content (mg − 1 g 1 FW) 48 hrs 96 hrs 144 hrs 240 hrs Treated Control Treated Control Treated Control Treated Control GLK 10–40 9.508 4.980 11.899 4.473 13.119 4.582 6.293 4.377 GLK 20055 8.203 4.364 9.437 4.763 11.408 4.340 5.987 4.784 FLIP-09-194C 7.129 5.812 8.208 5.058 13.210 5.644 6.591 5.724 FLIP-04-219C 8.485 4.601 9.028 4.173 14.310 4.438 7.164 4.919 ICCV 55215 10.352 4.862 10.694 4.922 12.085 4.193 5.697 4.328 GLK 17301 3.023 2.516 3.346 2.544 4.992 2.510 2.224 2.101 C.D. 0.575 0.958 1.542 1.075 SE(m) 0.185 0.307 0.495 0.345 SE(d) 0.261 0.435 0.700 0.488 C.V. 4.148 6.505 8.608 9.763 4. Discussion Ascochyta blight poses a significant threat to chickpea cultivation, particularly in areas characterized by cool, cloudy, and humid weather throughout the crop season. Studies indicate that under favorable epidemic conditions, ascochyta blight can lead to complete yield loss in chickpea crops (Bahr et al 2016 ). Research on plant-pathogenic interactions involving fungi, bacteria, and viruses has highlighted the toxicity of specific common phenols and phenolic substances towards pathogens. These compounds, traditionally recognized as crucial defense-related elements, are naturally abundant in resistant varieties of various crops (Gogoi et al., 2001 ). Moreover, they tend to accumulate in plants following infection, particularly in resistant varieties. PAL is the first enzyme in the phenylpropanoid pathway that leads to the formation of secondary metabolites. Its direct role in lignification under stress conditions or infections caused by pathogens is well known. The observed increase in PAL activity till 96 hours after inoculation, particularly in resistant lines, underscores the enzyme's pivotal role in the early defense response against Ascochyta blight. Resistant genotypes, notably GLK 10–40, exhibited sustained and heightened PAL activity, indicating an effective defense mechanism. Our study aligns with the findings of Manjunatha et al. ( 2018 ), indicating that the increase in PAL activity was more pronounced in the resistant genotype (ICCV 5530) compared to the susceptible ones. This observation suggests a robust defense response in ICCV 5530 against Ascochyta rabiei. Tyrosine ammonia-lyase (TAL) is a key enzyme in plant defense, converting tyrosine into coumaric acid. It operates in the phenylpropanoid metabolic pathway, essential for producing secondary metabolites like phenolic compounds and lignin (Sindhu et al., 1995 ). Similar to PAL, the elevation in TAL activity until 96 hours after inoculation suggests its involvement in the initial defense cascade against fungal infection. Resistant lines, especially GLK 20055, displayed higher and sustained TAL activity, indicating its potential as a key player in defense mechanisms. Polyphenol oxidase (PPO) is a vital component of the plant's immune system, generating defensive compounds. In response to injuries or pathogen attacks, plants release chemical signals, triggering the activation of PPO. This enzyme catalyzes the oxidation of phenolic compounds, resulting in the formation of highly reactive quinones (Mayer et al., 1987). The increase in PPO activity until 96 hours after inoculation highlights its crucial role in the early defense response against Ascochyta blight. Resistant genotypes, with GLK 10–40 showing peak activity on the 4th day, exhibited higher PPO levels, suggesting its association with resistance. The observed decline in PPO activity after the peak indicates a controlled defense strategy. Kaur et al ( 2021 ) found that (PPO) resulted in reduced root rot and wilt symptoms in the resistant cultivar as compared to the susceptible cultivar. Results showed that banana fruit irradiated with UVC at 24h after fungal. The rise in POD activity until 96 hours after inoculation, particularly in resistant lines like GLK 20055, signifies its involvement in the early and middle phases of the defense against Ascochyta blight. The sustained POD activity in resistant lines suggests its importance in prolonged defense mechanisms. The results are in line with Nawar and Kuti ( 2003 ), who proposed that increased peroxidase activity is an early sign of resistance to chocolate spot disease in broad beans, serving as a protective barrier against pathogens. This aligns with the defense mechanisms involving protein accumulation and peroxidase against plant pathogens (Sarwar et al., 2011 ). Furthermore, Hassan et al. ( 2007 ) found that elevated peroxidase activity in faba beans, induced by certain molecules, indicates resistance to chocolate spot disease. Lignin, a complex polymer in plant cell walls, provides structural support and acts as a physical barrier in plant defense mechanisms against pathogens. While effective against many pathogens, some specialized ones can overcome or degrade lignin barriers (Kamran et al. 2018 ).The increase in lignin content until the 6th day emphasizes its role as a structural component in the defense against Ascochyta blight. Genotypes, particularly GLK 20055, consistently exhibiting higher lignin content, suggest its significance in resistance. A similar result was found by Egea et al. ( 2001 ), who observed that the basal levels of lignin were higher in the S-5 resistant variety compared to the other two susceptible varieties studied. Total phenols resist microbial infection by inactivating fungal enzymes, accumulating at the injury or penetration site to inhibit phytopathogen growth. Phenols, widely distributed in crop plants, contribute to resistance mechanisms against parasitic or pathogenic fungi (Dogan et al., 2007 ). The elevation in total phenol content until the 6th day highlights their significance in the defense against Ascochyta blight. Resistant genotypes, notably FLIP-04-219C, exhibiting higher and sustained total phenol content, suggest the potential role of phenolic compounds in defense mechanisms. Manjunatha et al ( 2018 ) findings align with our study, indicating that the resistant genotype ICCV 5530 consistently exhibited higher phenol levels in both healthy and inoculated conditions at 2, 4, and 8 days after inoculation. In contrast, the susceptible genotype ICCV 4991 demonstrated a decrease in total phenol content throughout the inoculation stages. Manipulating PAL, TAL, and PPO activity through genetic engineering or targeted breeding, with GLK 10–40 as a promising candidate, POD and lignin content with line GLK 20055, and total phenol content with genotypes like FLIP-04-219C, could lead to chickpea cultivars with improved resistance to Ascochyta blight. 5. Conclusion In the effort to strengthen chickpea resistance against the challenging Ascochyta blight, this study explored the intricate biochemical aspects that govern plant defense mechanisms. Investigating the dynamic shifts of enzyme activities and biochemical components post-fungal infection has provided valuable insights, offering promising directions for improving resistance strategies.The identified genotypes, particularly GLK 10–40, GLK 20055, and FLIP-04-219C, emerged as most prominent resistant cultivars, showcasing increased enzymatic activities crucial for early defense against Ascochyta blight. PAL, TAL, PPO, and POD enzymes, along with lignin and total phenols, played pivotal roles in the facilitation of a complex defense mechanism. The surge in PAL, TAL, PPO, and POD activities during the critical 96-hour window underscored their importance in the initial defense cascade. Lignin, elevating consistently until the 144 hour, and total phenols, maintaining increased levels, highlighted their structural and non-enzymatic contributions to resisting Ascochyta blight.As we traverse beyond conventional resistance mechanisms, the findings unravel opportunities for targeted breeding or genetic engineering. By leveraging the natural defense arsenal within genotypes like GLK 10–40, GLK 20055, and FLIP-04-219C, the quest for resistant chickpea cultivars gains momentum. This research not only contributes to the scientific understanding of host-pathogen interactions but also holds practical implications for global food security References Angelini R, Bragaloni M, Federico R, Infantino A, and Porta-Pugua A (1993) Involvement of polyamines, diamine oxidase and peroxidase in resistance of chickpea to Ascochyta rabiei. J Pl Physiol 142: 704-09. Bahr, L., Castelli, M.V., Barolo, M.I., Mostacero, N.R., Tosello, M.E. and López, S.N., 2016. Ascochyta blight: isolation, characterization, and development of a rapid method to detect inhibitors of the chickpea fungal pathogen Ascochyta rabiei. Fungal biology , 120 (3), pp.424-432. Chandra-Hioe, M.V., Wong, C.H. and Arcot, J., 2016. The potential use of fermented chickpea and faba bean flour as food ingredients. Plant Foods for Human Nutrition , 71 , pp.90-95. 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H., Ashfaq, S., & Jamil, F. F. (2011). Induced systemic resistance in chickpea against Ascochyta blight by safe chemicals. Pakistan Journal of Botany , 43 (2), 1381-1387. Sindhu A N I L, Singh R A J E N D E R, Nehra K S and Singal H R (1995) Elicitor-induced metabolic changes in seedlings of chickpea (Cicer arietinum L.) in relation to Ascochyta blight. Annals of Biology 11: 173-83. Wise, K.A., Bradley, C.A., Markell, S., Pasche, J., Delgado, J.A., Goswami, R.S. and Gudmestad, N.C., 2011. Sensitivity of Ascochyta rabiei populations to prothioconazole and thiabendazole. Crop Protection , 30 (8), pp.1000-1005. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ekabuli\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003echickpea lines in both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs post-inoculation with \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAscochyta rabiei\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-3886689/v1/d8b98bc886a47207a77df27c.png"},{"id":50372667,"identity":"37b2982a-b691-4cc0-ab5b-32a3709a0530","added_by":"auto","created_at":"2024-01-30 13:46:42","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":62540,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDifferent range of intervals between the maximum and minimum change in the activity of Lignin in resistant and highly susceptible \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ekabuli\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e chickpea lines in both treated/inoculated and control/untreated lines at 48, 96, 144 and 240 hrs post-inoculation with \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAscochyta rabiei\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-3886689/v1/0a1abe778b5ee91ad1309ece.png"},{"id":50373151,"identity":"f5519719-fea5-47b4-ae31-aff916565087","added_by":"auto","created_at":"2024-01-30 13:54:42","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":69679,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eChanges in lignin content in individual resistant and highly susceptible \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ekabuli\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e chickpea lines in both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs post-inoculation with \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAscochyta rabiei\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-3886689/v1/b8ae565da45c6ab1590ebabc.png"},{"id":50372669,"identity":"c26efd63-55a4-4175-9dde-279494a9a0b2","added_by":"auto","created_at":"2024-01-30 13:46:42","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":46279,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDifferent range of intervals between the maximum and minimum change in the activity of Total Phenols in resistant and highly susceptible \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ekabuli\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e chickpea lines in both treated/inoculated and control/untreated lines at 48, 96, 144 and 240 hrs post-inoculation with \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAscochyta rabi\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","description":"","filename":"11.png","url":"https://assets-eu.researchsquare.com/files/rs-3886689/v1/14019003fd9ac7de17297428.png"},{"id":50372670,"identity":"b3d46122-8098-42e0-9ccc-9263455cb738","added_by":"auto","created_at":"2024-01-30 13:46:42","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":58255,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eChanges in Total Phenols in individual resistant and highly susceptible \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ekabuli\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e chickpea lines in both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs post-inoculation with \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAscochyta rabiei\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"12.png","url":"https://assets-eu.researchsquare.com/files/rs-3886689/v1/21536609288c2707052bc77a.png"},{"id":64870107,"identity":"99d848e0-de91-4cb7-87ed-dedb0d78eb14","added_by":"auto","created_at":"2024-09-19 21:00:58","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2566787,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3886689/v1/343e13a4-b7ee-45f4-9026-a462e8f9c8f0.pdf"}],"financialInterests":"","formattedTitle":"Defence mechanism through a biochemical pathway in Kabuli chickpea genotypes against Ascochyta blight","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eChickpea (\u003cem\u003eCicer arietinum\u003c/em\u003e L.) holds significance as the second-largest pulse crop, cultivated and consumed worldwide, predominantly in Afro-Asian nations. Global chickpea production reached approximately 15.87\u0026nbsp;million tons with an area of 16.01\u0026nbsp;million hectare in 2022. In India, chickpea cultivation covers an area of approximately 10.94\u0026nbsp;million hectares resulting in an annual production of 11.91\u0026nbsp;million tons (FAOSTAT \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Chickpeas constitute a valuable reservoir of crucial minerals such as zinc, iron, calcium, copper, potassium, and phosphorus, in addition to containing vitamin B and dietary fiber. This elevated dietary fiber content is especially beneficial for those with insulin sensitivity or diabetes (Pittaway et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Chickpea genotypes can be categorized into two main types: desi and kabuli. This classification is primarily determined by seed size, shape, and color. Desi chickpeas are predominantly cultivated in Asia and Africa, whereas kabuli chickpeas are commonly found in the Mediterranean region and are extensively grown in North America, particularly in Mexico and the United States (Chandra-Hioe et al \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). While chickpea production has seen recent growth, it still struggles to meet the demands of an increasing vegetarian population. The challenge lies in enhancing production and productivity, given the limitations in expanding cultivable areas. Factors like human intervention, evolving cultural practices, and climate change contribute to an increase in diseases, affecting crop yields. A total of 172 chickpea pathogens have been reported globally, including approximately 67 fungi, 3 bacteria, 22 viruses, and 80 nematodes, posing significant biotic stresses on chickpea cultivation (Nene et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1996\u003c/span\u003e). However, only a handful of these pathogens can cause widespread damage, leading to epidemics that significantly impact global chickpea production. Diseases like Ascochyta blight, Botrytis gray mold, Fusarium wilt, dry root rot, black root rot, collar rot, Phytophthora, Pythium root rot, seed rot, and seedling rot pose a major economic threat. Others have sporadic occurrences and localized distributions (Johansen et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1994\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAschochyta blight by the highly virulent necrotrophic pathogen \u003cem\u003eAscochyta rabiei\u003c/em\u003e (Pass.) Labr. (syn. \u003cem\u003eDidymella rabiei\u003c/em\u003e) in chickpeas (Owati et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). This pathogen can infect crops at various stages, leading to significant yield reduction, particularly during flowering and podding stages (Wise et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The disease is more prevalent under cool and wet conditions, with temperatures ranging from 15 to 25\u0026deg;C and relative humidity between 65% and 100%, favoring its development and spread. Severe infections result in reduced seed quality and complete yield loss (Pande et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Aschochyta blight manifests as lesions on foliar and stem parts, leading to stem breakage and seed rot. Symptomatic features include bended or lengthened darker brown-red lines on leaflets, concentric circles of pycnidia on leaves, stems, and pods, and 3\u0026ndash;4 cm length brown lesions with black spots on stems and petioles (Li et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eModern cultivars frequently lose resistance and become susceptible to diseases over time. The use of fungicides for disease management is often impractical and economically unfeasible (Reddy et al., 1990). The mechanisms underlying resistance during the host-pathogen interaction in both resistant and susceptible genotypes remain poorly understood. Plants, in response to pathogens, possess effective defense mechanisms where static and dynamic secondary metabolites play crucial roles as local or systemic resistance factors (Redman et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1999\u003c/span\u003e). The interaction between the host and pathogen induces signalling molecules in the plant system, leading to the production of antimicrobial phenolic substances. These substances participate in various biochemical processes. In certain diseases, infection is marked by an increased synthesis of phenolic compound precursors and oxidation products of phenolics, such as quinones, which demonstrate heightened toxicity to microorganisms (Bahr et al \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eUnderstanding biochemical defense mechanisms is crucial for defining resistance in specific genotypes. Induction of resistance in plants involves physiological and biochemical changes, including callose, phytoalexins, lignin, and plant pathogenesis-related proteins. Enzymes such as peroxidase (POD), phenylalanine ammonia-lyase (PAL), and polyphenol oxidase (PPO) play a vital role in the defense activities of plants against various abiotic and biotic inducers, contributing to resistance. In chickpeas, higher activity of PPO, PAL, and peroxidase, along with low catalase activity in leaf samples, appears to be a significant biochemical factor contributing to resistance against Ascochyta blight infections (Angelini et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1993\u003c/span\u003e). The understanding of these processes is essential for developing strategies to enhance plant resistance against diseases. Thus, this study significantly contributes to enhancing strategies for the effective management of Ascochyta blight. Through the identification of novel resistant sources in Kabuli chickpea and a deeper understanding of the biochemical defense mechanisms involved, our findings provide valuable insights that can be applied in developing robust and sustainable approaches to combat this detrimental disease.\u003c/p\u003e"},{"header":"2. Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Plant material and pathogen\u003c/h2\u003e \u003cp\u003eA total of 238 different Kabuli chickpea germplasm lines were screened against Ascochyta blight (\u003cem\u003eAscochyta rabiei\u003c/em\u003e) under artificial epiphytotic conditions during the rabi seasons of 2019-20, 2020-21 and 2021-22 at the Research Farms of the Pulses Section, Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana. Out of the 238 lines, six lines namely, GLK 10\u0026ndash;40, GLK 20055, FLIP 09-194C, FLIP 04-219C, ICCV 55215, and GLK 17301were chosen based on exhibiting both resistant and susceptible reactions (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Two sets, comprising five resistant and one susceptible line each, were sown. In one set, the lines were inoculated with \u003cem\u003eAscochyta rabiei\u003c/em\u003e local isolate using the foliar spray method, while the other set remained uninoculated (control). Artificial raining effect and relative humidity were maintained with perfo-spray system for disease development during the daytime from 10.00 am to 04:00 pm regularly for 21 days. Sampling was done at intervals of 48, 96, 144, and 244 hours after inoculation. Collected samples were placed in an icebox and promptly stored at -20\u0026deg;C for subsequent enzymatic analysis, including phenylalanine ammonia lyase (PAL), tyrosine ammonia lyase (TAL), and polyphenol oxidase (PPO), peroxidase (POD). For the estimation of lignin and total phenols, samples were gathered in an icebox and preserved in methanol (80%) for total phenols and ethanol (90%) for lignin, both stored at -20\u0026deg;C.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eResistant and susceptible \u003cem\u003ekabuli\u003c/em\u003e chickpea lines against Aschochyta blight (\u003cem\u003eAschochyta rabiei\u003c/em\u003e) for biochemical analysis\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eS.No\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLine Name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDisease Score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eReaction Type\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGLK 10\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eResistance\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGLK 20055\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eResistance\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFLIP 09-194C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eResistance\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFLIP 04-219C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eResistance\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eICCV 55215\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eResistance\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGLK 17301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSusceptible\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Disease severity scale\u003c/h2\u003e \u003cp\u003eThe data were collected using a rating scale of 1\u0026ndash;9 points, as outlined in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, and further detailed in instances where the susceptibility check revealed maximum disease. The lines were classified as highly resistant (HR), resistant (R), moderately resistant (MR), susceptible (S), or highly susceptible (HS) based on this rating scale (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eRating scale (1\u0026ndash;9) against \u003cem\u003eAscochyta\u003c/em\u003e blight of chickpea\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDisease score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDescription\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eReaction types\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo visible symptom\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHighly Resistant (HR)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSmall/minute lesions were observed on foliage of plants, mostly not visible\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eResistant (R)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFew scattered lesions were observed on foliage and stems of plants, mostly visible after careful examination\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLesions were detected on foliage and stems of few plants but were not damaging.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eModerately resistant (MR)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLesions were commonly observed on foliage and stems but damage was less\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLesions commonly detected on foliage and stems; 25.0% or one fourth of the plants would be killed.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSusceptible(S)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLesions would be commonly observed on foliage and stems, 50.0% or half of the plants would be killed.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLesions were extensively observed on all of the plants, girdling of stem and branches visible, 75.0% or three fourth of the plants would be killed.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eHighly Susceptible (HS)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLesions were extensively observed on all of the plants tissues, girdling was extensive on all of the plant parts including stems and branches and \u0026gt;\u0026thinsp;75.0% or more than three fourth of plants would be killed.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Extraction and estimation of enzymes\u003c/h2\u003e \u003cp\u003eThe enzymatic antioxidants such as phenylalanine ammonia lyase (PAL), tyrosine ammonia lyase (TAL), polyphenol oxidase (PPO), peroxidase (POD) lignin, and total phenols were estimated in twigs of chickpea at four stages such as S1 (48 hours), S2 (96 hours), S3 (144 hours), and S4 (240 hours). To determine enzymatic antioxidants, the enzymes were extracted using extraction buffers at 4\u0026deg;C to minimize enzyme denaturation, and the assays were conducted at 30\u0026deg;C.\u003c/p\u003e \u003cp\u003eFor extraction of PAL and TAL enzymes, 0.1 g of fresh chickpea plant samples were homogenized in about 2 ml of Tris HCl (0.1 M) buffer having a pH of 7.5 and containing β-mercaptoethanol (5 mM). Subsequently, they were centrifuged for 25 m at temperature of 4\u0026ordm;C and 10,000 rpm. Then, the supernatant was utilized to estimate the PAL and TAL. For estimating the activity of PAL enzyme the reaction mixture containing 2.5 ml of 30 mM phenylalanine prepared in 0.05 M sodium borate buffer (pH 8.8) and 100 \u0026micro;l of enzyme extract was incubated at 37\u0026ordm;C for 1 hr. The reaction was stopped by the addition of 0.3 ml of 5 N HCl. The absorbance was recorded at 290 nm against blank (simple water). No incubation was done in case of control treatment and the reaction was stopped right away. Additionally, 5\u0026ndash;40 \u0026micro;g of cinnamic acid was utilized to create a standard curve that was further used to calculate the activity of PAL. Furthermore, the activity was expressed as \u0026micro;g of cinnamic acid formed min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003emg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e fresh weight, whereas for in case of TAL enzyme the reaction mixture for TAL was also incubated at 37\u0026ordm;C for 1 hour and contained 1 ml of 33 \u0026micro;M tyrosine, 50 \u0026micro;l of enzyme extract and also sodium borate buffer (1.35 ml) having a pH of 8.8. Furthermore, the reaction was stopped by the addition of 5 N HCl (0.1 ml). The absorbance was recorded at 310 nm against the water blank. As there was no incubation in case of control, the reaction was stopped right away. The standard curve made with coumaric acid (5\u0026ndash;40 \u0026micro;g) was used to calculate the activity of TAL. Its activity was further expressed as \u0026micro;g of coumaric acid formed min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003emg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e fresh weight.\u003c/p\u003e \u003cp\u003eFor extraction and estimation of PPO enzyme activity Freshly weighed chickpea plant samples (0.1g) were crushed in 2 ml of ice-cooled 0.1 M potassium phosphate buffer (pH 6.8) containing 2-mercaptoethanol (5 mM), 1% PVP and 1 mM EDTA before being centrifuged at 10,000 rpm for 20 min. The supernatant collected was used as an enzyme extract. The reaction mixture consisted of 100 mM 4-methyl catechol, buffer and enzyme extract. The change in absorbance was recorded at 410 nm for 3 min at an interval of 30 sec. One unit of enzyme activity was defined as an increase of 0.01 in absorbance min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003eof fresh weight.\u003c/p\u003e \u003cp\u003eFor extraction and estimation of POD enzyme activity freshly weighed chickpea plant samples (0.1 g) were homogenized in 2 ml of ice-cold 0.1 M potassium phosphate buffer (pH 7.0) containing 2 β-mercaptoethanol (5 mM), 1% PVP and EDTA. The material was centrifuged at 10,000 rpm for 20 min after being filtered through muslin cloth. An enzyme extract was prepared using the collected supernatant. The supernatant was collected and used as enzyme extract. The reaction mixture consisted of 0.05 M guaiacol, 100 \u0026micro;l enzyme extract and 0.8 M H₂O₂. The change in absorbance was recorded at 470 nm for 3 min at an interval of 30 sec using the UV-visible spectrophotometer. The enzyme activity of peroxidase was expressed as a change in absorbance min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e mg\u003csup\u003e1\u003c/sup\u003eof fresh weight.\u003c/p\u003e \u003cp\u003eFor extraction and estimation of lignin content fresh plant samples (0.1 g) of treated and untreated chickpea plants were homogenized in 2 ml of 95% ethanol. The homogenate was then centrifuged at 10,000 rpm for 20 min. The supernatant was discarded and the pellet was washed thrice with 95% ethanol and twice with freshly prepared ethanol: hexane (1:2) solution. Washed pellets were dried overnight at 45\u0026deg;C. After being dissolved in 25% acetyl bromide made in acetic acid, the dried pellet was incubated at 70\u0026deg;C for 30 min before being cooled at ambient temperature. The extract so obtained was used for estimating the lignin content. The reaction mixture consisted of 100 \u0026micro;l extract, 180 \u0026micro;l of 2 N NaOH, 20 \u0026micro;l of 7.5 M NH₂OH.HC1 and 1.6 ml acetic acid. After centrifuging this mixture for 5 min, the intensity of colour generated was measured at 280 nm. The simultaneous running lignin standard curve (5\u0026ndash;40 \u0026micro;g) was used to calculate the lignin content. The, so obtained, lignin content was expressed as mg g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003efresh weight.\u003c/p\u003e \u003cp\u003eFor extraction and estimation of total phenol content fresh chickpea plant samples (300 mg) were homogenized in 3 ml of 80% methanol and refluxed with methanol for 1 hr. The refluxed content was filtered and the pellet was re-extracted with 80% methanol. The filtrate from these two extractions was then mixed and methanol was used to make up the volume to 5 ml. This methanolic extract was used to calculate the amount of total phenols. Methanolic extract (0.5 ml) was taken in test tubes and evaporated to dryness. To this, 6.5 ml distilled water and 0.5 ml Folin's-Phenol reagent (prepared in 1:1 with distilled water) were added and vortexed to mix well. Then 1.0 ml saturated solution of sodium carbonate was added and the mixture was kept at room temperature for one hour. The intensity of blue colour developed was read spectrophotometrically at 760 nm against the reagent blank. The standard curve was prepared using gallic acid (10\u0026ndash;50 \u0026micro;g). Total phenols, so obtained, were expressed as mg of gallic acid equivalent 100 g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e fresh weight.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Result","content":"\u003cp\u003eIn the current study, six Kabuli chickpea genotypes were cultivated under both controlled and infected conditions. These genotypes were classified according to disease incidence, with GLK 10\u0026ndash;40, GLK 20055, FLIP 09-194C, FLIP 04-219C, and ICCV 55215 identified as resistant genotypes, and GLK 17301 designated as a susceptible genotype (refer to Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Biochemical parameters were assessed in the leaves of these genotypes at four stages and various hours after inoculation (HAI).\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Biochemical reactions in response to the occurrence of Ascochyta blight\u003c/h2\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e3.1.1 Phenylalanine Ammonia-Lyase (PAL)\u003c/h2\u003e \u003cp\u003ePAL enzyme activity in response to \u003cem\u003eAscochyta rabiei\u003c/em\u003e inoculation in chickpea genotypes, with results indicating an increase in activity in resistant chickpea lines (GLK 10\u0026ndash;40, GLK 20055, FLIP-09-194C, FLIP-04-219C, and ICCV 55215) displayed higher PAL enzyme activity than the susceptible line (GLK 17301), peaking at 96 hrs and declining thereafter in treated lines(Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Treated plants of GLK 10\u0026ndash;40 displayed highest PAL activity (33.06 \u0026micro;g cinnamic acid formed min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW at 96 hrs and 18.612 \u0026micro;g cinnamic acid formed min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW at 144 hrs) and significant increase of 45.5-fold at 96 hrs followed by 26.40-fold rise at 144 hrs. Conversely, treated GLK 17301(susceptible genotype) consistently showed lower mean PAL activity (2.52 \u0026micro;g cinnamic acid formed min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) than resistant treated lines with considerable increases compared to control treatments at different tested time intervals (Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePhenylalanine ammonia-lyase (PAL) activity in \u003cem\u003ekabuli\u003c/em\u003e chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with \u003cem\u003eAscochyta rabiei\u003c/em\u003e.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eName\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"8\" nameend=\"c9\" namest=\"c2\"\u003e \u003cp\u003ePhenylalanine Ammonia Lyase (PAL) activity (\u0026micro;g cinnamic acid formed min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e48 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e96 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e144 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e240 hrs\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGLK 10\u0026ndash;40\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e6.503\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e0.722\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e33.063\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e0.723\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e18.612\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.705\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.987\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e0.705\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGLK 20055\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e4.356\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e0.465\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e18.374\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e0.460\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e11.215\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.431\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.706\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e0.529\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eFLIP-09-194C\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e7.658\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e0.542\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e21.729\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e0.583\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e8.202\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.504\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.601\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e0.462\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eFLIP-04-219C\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e3.932\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e0.384\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e12.338\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e0.303\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e8.708\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.375\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.497\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e0.365\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eICCV 55215\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e4.872\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e0.360\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e12.128\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e0.340\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e7.755\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.372\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.511\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e0.374\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGLK 17301\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e1.218\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e0.158\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e6.674\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e0.188\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e2.005\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.163\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.225\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e0.189\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eC.D.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e\u003cb\u003e1.088\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cb\u003e2.881\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u003cb\u003e1.627\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.183\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSE(m)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e\u003cb\u003e0.349\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cb\u003e0.925\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u003cb\u003e0.522\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.059\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSE(d)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e\u003cb\u003e0.494\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cb\u003e1.308\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u003cb\u003e0.738\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.083\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eC.V.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e\u003cb\u003e18.828\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cb\u003e13.21\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u003cb\u003e13.528\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e\u003cb\u003e14.118\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003e3.1.2 \u003cb\u003eTyrosine Ammonia-Lyase\u003c/b\u003e\u003c/h2\u003e \u003cp\u003eTAL is also vital enzyme of defense pathways and results showed increased TAL activity at 96 hrs post-inoculation which started decreasing afterwards. Comparisons revealed significant increase in TAL activity in all treated genotypes. Treated resistant line, GLK 20055 exhibited the highest activity (68.75 \u0026micro;g coumaric acid formed min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) at 96 hrs while susceptible genotype, GLK 17301 had the lowest (1.68 \u0026micro;g coumaric acid formed min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) at 240 hrs (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Control lines of both resistant and susceptible genotypes maintained constant TAL activity. FLIP-09-194C (resistant line) had the maximum TAL activity at 48 (6.57-fold increase) and 144 (10.72-fold rise) hrs respectively while GLK 20055 (resistant line) reached the highest value at 96 and 240 hrs with 15.74-fold and 2.49-fold rise respectively. The susceptible treated genotype, GLK 17301 consistently showed lower average TAL activity (4.91 \u0026micro;g coumaric acid formed min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) as compared to resistant treated lines with slight increases at different time intervals compared to controls. These findings depict dynamic changes in TAL enzyme activity in response to fungal infection in different chickpea genotypes, with the time frame now expressed in hours (Figs.\u0026nbsp;3 and \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTyrosine ammonia-lyase (TAL) activity activity in \u003cem\u003ekabuli\u003c/em\u003e chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with \u003cem\u003eAscochyta rabiei\u003c/em\u003e.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eName\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"8\" nameend=\"c9\" namest=\"c2\"\u003e \u003cp\u003eTyrosine Ammonia Lyase (TAL) activity (\u0026micro;g Coumaric acid formed min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e48 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e96 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e144 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e240 hrs\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 10\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15.225\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.441\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e52.250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.157\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e22.010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.794\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e8.333\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e6.922\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 20055\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e17.897\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.034\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e68.750\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.368\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e29.113\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.475\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9.873\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.951\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFLIP-09-194C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18.054\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.745\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e33.216\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.990\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e29.373\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.740\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.510\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.833\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFLIP-04-219C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15.971\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.059\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e29.667\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.373\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e22.328\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.676\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4.637\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.417\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eICCV 55215\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15.873\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.377\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e26.039\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.078\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e17.456\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.637\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6.505\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e4.265\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 17301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.887\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.691\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.868\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.623\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.216\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.539\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.686\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1.544\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.D.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e1.423\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e5.795\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e3.658\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e0.895\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSE(m)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e0.457\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e1.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e1.174\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e0.287\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSE(d)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e0.646\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e2.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e1.661\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e0.406\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.V.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e6.947\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e14.364\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e13.406\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e8.787\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eFig 3. Different range of intervals between the maximum and minimum change in the activity of Tyrosine ammonia-lyase (TAL) in resistant and highly susceptible\u003c/b\u003e \u003cb\u003ekabuli\u003c/b\u003e \u003cb\u003echickpea lines in both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs post-inoculation with\u003c/b\u003e \u003cb\u003eAscochyta rabiei\u003c/b\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003e3.1.3 Polyphenol Oxidase (PPO)\u003c/h2\u003e \u003cp\u003eResults of PPO enzyme revealed increased activity till 96 hrs after inoculation, reaching the maximum limit and then declining. GLK 10\u0026ndash;40 (resistant) showed the highest PPO activity (345.83 unit\u0026rsquo;s min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) at 96 hrs while GLK 17301 (susceptible) had the lowest (52.25 unit\u0026rsquo;s min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) at 240 hrs in treated genotypes (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Control lines maintained constant PPO activity. PPO activity was highest in resistant treated genotypes i.e. GLK 20055 (233.50 units min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) at 48 hrs, GLK 10\u0026ndash;40 at 96 (345.83 units min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) and 144 (275.50 units min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) hrs respectively and ICCV 55215 (93.50 units min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) on 240 hrs. Comparisons showed significant increases in PPO activity in treated plants with a 5.24-fold rise in GLK 10\u0026ndash;40 at 96 hrs. The treated susceptible genotype, GLK 17301 consistently exhibited mean lower PPO activity (119.59 unit\u0026rsquo;s min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW) with significant increases at different time intervals compared to controls. These findings depict dynamic changes in PPO enzyme activity in response to fungal infection in different chickpea genotypes, with the timeframe expressed in hours (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e5\u003c/span\u003e and \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePolyphenol oxidase (PPO) activity activity in \u003cem\u003ekabuli\u003c/em\u003e chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with \u003cem\u003eAscochyta rabiei\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eName\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"8\" nameend=\"c9\" namest=\"c2\"\u003e \u003cp\u003ePolyphenol oxidase (PPO) units min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e48 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e96 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e144 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e240 hrs\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 10\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e156.250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e63.333\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e345.833\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e66.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e275.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e64.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e76.250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e63.750\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 20055\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e233.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e74.417\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e311.917\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e78.167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e271.250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e76.917\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e88.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e77.583\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFLIP-09-194C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e166.667\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e68.083\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e289.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e66.250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e244.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e61.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e76.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e68.417\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFLIP-04-219C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e151.167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e61.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e237.750\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e66.417\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e217.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e63.667\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e74.167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e58.500\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eICCV 55215\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e177.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e67.333\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e254.833\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e67.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e215.250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e64.250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e93.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e65.750\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 17301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e102.083\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e166.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e46.583\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e157.500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e47.833\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e52.250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e47.500\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.D.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e14.438\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e21.664\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e13.813\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e8.792\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSE(m)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e4.634\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e6.954\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e4.434\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e2.822\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSE(d)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e6.554\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e9.834\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e6.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e3.991\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.V.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e6.202\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e5.909\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e4.316\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e5.796\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section3\"\u003e \u003ch2\u003e3.1.4 Peroxidase (POD)\u003c/h2\u003e \u003cp\u003ePeroxidase enzyme (POD) prevents accumulation of hydrogen peroxide by catalyzing different substrates, however this study indicated that its activity increased after 96 hrs of inoculation in all treated plants with the highest enzyme activity observed in the resistant plants. The maximum enzymatic activity was observed resistant genotype, GLK 20055 at 96 (247.418 ∆A min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW) hrs, followed by 144 \u003cb\u003e(\u003c/b\u003e197.948 ∆A min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW), 48\u003cb\u003e(\u003c/b\u003e183.678 ∆A min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW) and 240 (97.290 ∆A min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW) hrs respectively while GLK 17301 genotype showed the lowest on 240 (49.92 ∆A min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW) hrs (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). The control plants maintained a consistent level of POD activity. Comparisons between treated and control plants revealed significant increases in POD activity in treated resistant genotype, GLK 20055 showing 8.2-fold increase at 96 hrs, whereas the susceptible treated genotype GLK 17301 exhibited average lower POD activity(62.23 ∆A min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW), with significant increases at different time intervals compared to controls. These findings illustrate dynamic changes in POD enzyme activity in response to fungal infection in different chickpea genotypes, with the timeframe expressed in hours (Figs.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e7\u003c/span\u003e and \u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e8\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePeroxidase (POD) activity activity in \u003cem\u003ekabuli\u003c/em\u003e chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with \u003cem\u003eAscochyta rabiei.\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eName\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"8\" nameend=\"c9\" namest=\"c2\"\u003e \u003cp\u003ePeroxidase activity (POD) (∆A min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e48 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e96 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e144 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e240 hrs\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 10\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e150.253\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e58.143\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e213.600\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e50.667\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e194.900\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e51.650\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e93.195\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e51.925\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 20055\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e183.678\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e39.900\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e247.418\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e30.143\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e197.9483\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e36.292\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e97.290\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e34.075\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFLIP-09-194C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e150.737\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e44.682\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e235.860\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.438\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e183.200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e47.817\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e96.045\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e42.587\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFLIP-04-219C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e181.157\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75.900\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e198.055\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.758\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e195.360\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e77.013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e96.622\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e60.960\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eICCV 55215\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e166.675\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e40.153\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e209.120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e44.050\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e172.773\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e47.657\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e71.925\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e45.573\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 17301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e65.705\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e31.765\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e67.633\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e26.458\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e65.670\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e30.938\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e49.923\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e31.917\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.D.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e15.484\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e9.429\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e11.405\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e11.995\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSE(m)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e4.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e3.026\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e3.661\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e3.85\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSE(d)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e7.029\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e4.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e5.177\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e5.445\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.V.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e6.917\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e3.548\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e4.678\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e8.239\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003e3.1.5 Lignin content\u003c/h2\u003e \u003cp\u003ePlant defence component lignin increased till 144 hrs post-inoculation in all treated lines, with higher levels in resistant lines. GLK 20055 (resistant genotype) had significantly maximum lignin content at different time intervals (48, 96, 144 and 240 hrs), while minimum lignin content was observed in GLK 17301(susceptible genotype) (Table\u0026nbsp;\u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e). Comparisons revealed noteworthy increase in lignin content in treated resistant genotype, GLK 20055 with 4.79-fold rise at 144 hrs, whereas susceptible treated genotype, GLK 17301 consistently exhibited lower lignin content compared to controls at different time periods. These findings illustrate dynamic changes in lignin content in response to fungal infection in different chickpea genotypes, with the timeframe expressed in hours (Figs.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e9\u003c/span\u003e and \u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e10\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab7\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eLignin content in \u003cem\u003ekabuli\u003c/em\u003e chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with \u003cem\u003eAscochyta rabiei.\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eName\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"8\" nameend=\"c9\" namest=\"c2\"\u003e \u003cp\u003eLignin content (mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e48 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e96 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e144 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e240 hrs\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 10\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.630\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.787\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.130\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e11.158\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4.190\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.267\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 20055\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.966\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.158\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.585\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.009\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e14.947\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.174\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e5.253\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.218\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFLIP-09-194C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.090\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.907\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.651\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.131\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.537\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.851\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.708\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFLIP-04-219C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.371\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.657\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.188\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.734\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.422\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.328\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.769\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.397\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eICCV 55215\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.502\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.615\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.624\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.952\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.150\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.312\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4.102\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.766\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 17301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.827\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.135\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.019\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.041\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.796\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.143\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.241\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.025\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.D.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e0.834\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e1.304\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e0.547\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e0.581\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSE(m)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e0.268\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.418\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e0.176\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e0.187\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSE(d)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e0.379\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.592\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e0.248\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e0.264\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.V.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e10.106\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e12.126\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e4.193\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e7.674\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section3\"\u003e \u003ch2\u003e3.1.6 Total phenols\u003c/h2\u003e \u003cp\u003ePhenols provide structural integrity and support the plants defence mechanism by counteracting/nullifying pathogen attack. Increase in total phenol content was achieved until 144 hrs after inoculation in all treated lines, with higher levels in resistant lines compared to the susceptible line. The zenith of total phenol content in FLIP-04-219C (resistant treated genotype) of 14.310 mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW was achieved at 144 hrs and 7.164 mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW at 240 hrs respectively followed by resistant treated genotypes, GLK 10\u0026ndash;40 at 96 (11.899 mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW) hrs and ICCV 55215 at 48 (10.352 mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW) hrs respectively, while susceptible treated genotype, GLK 17301 showed minimum phenol content of 3.023 mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW at 48 hrs, 3.346 mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW at 96 hrs, 4.992 mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW at 144 hrs, and 2.224 mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW at 240 hrs post-inoculation respectively as compared to control lines (Table\u0026nbsp;\u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e8\u003c/span\u003e). Comparisons revealed significant increases in total phenol content in treated plants, notably a 4.22-fold rise in resistant line, FLIP-04-219C at 144 hrs but susceptible genotype, GLK 17301 exhibited lower average total phenol content (1.40-fold) with substantial increases at different time intervals compared to controls. These findings illustrate the dynamic changes in total phenol content in response to fungal infection in different chickpea genotypes, now expressed in hours (Figs.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e11\u003c/span\u003e and \u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e12\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab8\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTotal phenol content in \u003cem\u003ekabuli\u003c/em\u003e chickpea lines on both treated/ inoculated and control/untreated lines at 48, 96, 144 and 240 hrs after inoculation with \u003cem\u003eAscochyta rabiei\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eName\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"8\" nameend=\"c9\" namest=\"c2\"\u003e \u003cp\u003eTotal phenol content (mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e g\u003csup\u003e1\u003c/sup\u003e FW)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e48 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e96 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e144 hrs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e240 hrs\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 10\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.508\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.980\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11.899\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.473\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e13.119\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.582\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6.293\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e4.377\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 20055\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.203\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.364\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.437\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.763\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e11.408\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.340\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e5.987\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e4.784\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFLIP-09-194C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.129\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.812\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.208\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.058\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e13.210\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5.644\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6.591\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e5.724\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFLIP-04-219C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.485\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.601\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.028\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.173\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e14.310\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.438\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e7.164\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e4.919\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eICCV 55215\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.352\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.862\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10.694\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.922\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e12.085\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.193\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e5.697\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e4.328\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGLK 17301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.516\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.346\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.544\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.992\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.510\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.224\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.101\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.D.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e0.575\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.958\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e1.542\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e1.075\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSE(m)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e0.185\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.307\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e0.495\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e0.345\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSE(d)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e0.261\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.435\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e0.700\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e0.488\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.V.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e4.148\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e6.505\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e8.608\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003e9.763\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eAscochyta blight poses a significant threat to chickpea cultivation, particularly in areas characterized by cool, cloudy, and humid weather throughout the crop season. Studies indicate that under favorable epidemic conditions, ascochyta blight can lead to complete yield loss in chickpea crops (Bahr et al \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Research on plant-pathogenic interactions involving fungi, bacteria, and viruses has highlighted the toxicity of specific common phenols and phenolic substances towards pathogens. These compounds, traditionally recognized as crucial defense-related elements, are naturally abundant in resistant varieties of various crops (Gogoi et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). Moreover, they tend to accumulate in plants following infection, particularly in resistant varieties. PAL is the first enzyme in the phenylpropanoid pathway that leads to the formation of secondary metabolites. Its direct role in lignification under stress conditions or infections caused by pathogens is well known. The observed increase in PAL activity till 96 hours after inoculation, particularly in resistant lines, underscores the enzyme's pivotal role in the early defense response against Ascochyta blight. Resistant genotypes, notably GLK 10\u0026ndash;40, exhibited sustained and heightened PAL activity, indicating an effective defense mechanism. Our study aligns with the findings of Manjunatha et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), indicating that the increase in PAL activity was more pronounced in the resistant genotype (ICCV 5530) compared to the susceptible ones. This observation suggests a robust defense response in ICCV 5530 against \u003cem\u003eAscochyta rabiei.\u003c/em\u003e Tyrosine ammonia-lyase (TAL) is a key enzyme in plant defense, converting tyrosine into coumaric acid. It operates in the phenylpropanoid metabolic pathway, essential for producing secondary metabolites like phenolic compounds and lignin (Sindhu et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1995\u003c/span\u003e). Similar to PAL, the elevation in TAL activity until 96 hours after inoculation suggests its involvement in the initial defense cascade against fungal infection. Resistant lines, especially GLK 20055, displayed higher and sustained TAL activity, indicating its potential as a key player in defense mechanisms. Polyphenol oxidase (PPO) is a vital component of the plant's immune system, generating defensive compounds. In response to injuries or pathogen attacks, plants release chemical signals, triggering the activation of PPO. This enzyme catalyzes the oxidation of phenolic compounds, resulting in the formation of highly reactive quinones (Mayer et al., 1987). The increase in PPO activity until 96 hours after inoculation highlights its crucial role in the early defense response against Ascochyta blight. Resistant genotypes, with GLK 10\u0026ndash;40 showing peak activity on the 4th day, exhibited higher PPO levels, suggesting its association with resistance. The observed decline in PPO activity after the peak indicates a controlled defense strategy. Kaur et al (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) found that (PPO) resulted in reduced root rot and wilt symptoms in the resistant cultivar as compared to the susceptible cultivar. Results showed that banana fruit irradiated with UVC at 24h after fungal. The rise in POD activity until 96 hours after inoculation, particularly in resistant lines like GLK 20055, signifies its involvement in the early and middle phases of the defense against Ascochyta blight. The sustained POD activity in resistant lines suggests its importance in prolonged defense mechanisms. The results are in line with Nawar and Kuti (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2003\u003c/span\u003e), who proposed that increased peroxidase activity is an early sign of resistance to chocolate spot disease in broad beans, serving as a protective barrier against pathogens. This aligns with the defense mechanisms involving protein accumulation and peroxidase against plant pathogens (Sarwar et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Furthermore, Hassan et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2007\u003c/span\u003e) found that elevated peroxidase activity in faba beans, induced by certain molecules, indicates resistance to chocolate spot disease. Lignin, a complex polymer in plant cell walls, provides structural support and acts as a physical barrier in plant defense mechanisms against pathogens. While effective against many pathogens, some specialized ones can overcome or degrade lignin barriers (Kamran et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).The increase in lignin content until the 6th day emphasizes its role as a structural component in the defense against Ascochyta blight. Genotypes, particularly GLK 20055, consistently exhibiting higher lignin content, suggest its significance in resistance. A similar result was found by Egea et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2001\u003c/span\u003e), who observed that the basal levels of lignin were higher in the S-5 resistant variety compared to the other two susceptible varieties studied. Total phenols resist microbial infection by inactivating fungal enzymes, accumulating at the injury or penetration site to inhibit phytopathogen growth. Phenols, widely distributed in crop plants, contribute to resistance mechanisms against parasitic or pathogenic fungi (Dogan et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). The elevation in total phenol content until the 6th day highlights their significance in the defense against Ascochyta blight. Resistant genotypes, notably FLIP-04-219C, exhibiting higher and sustained total phenol content, suggest the potential role of phenolic compounds in defense mechanisms. Manjunatha et al (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) findings align with our study, indicating that the resistant genotype ICCV 5530 consistently exhibited higher phenol levels in both healthy and inoculated conditions at 2, 4, and 8 days after inoculation. In contrast, the susceptible genotype ICCV 4991 demonstrated a decrease in total phenol content throughout the inoculation stages. Manipulating PAL, TAL, and PPO activity through genetic engineering or targeted breeding, with GLK 10\u0026ndash;40 as a promising candidate, POD and lignin content with line GLK 20055, and total phenol content with genotypes like FLIP-04-219C, could lead to chickpea cultivars with improved resistance to Ascochyta blight.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eIn the effort to strengthen chickpea resistance against the challenging Ascochyta blight, this study explored the intricate biochemical aspects that govern plant defense mechanisms. Investigating the dynamic shifts of enzyme activities and biochemical components post-fungal infection has provided valuable insights, offering promising directions for improving resistance strategies.The identified genotypes, particularly GLK 10\u0026ndash;40, GLK 20055, and FLIP-04-219C, emerged as most prominent resistant cultivars, showcasing increased enzymatic activities crucial for early defense against Ascochyta blight. PAL, TAL, PPO, and POD enzymes, along with lignin and total phenols, played pivotal roles in the facilitation of a complex defense mechanism. The surge in PAL, TAL, PPO, and POD activities during the critical 96-hour window underscored their importance in the initial defense cascade. Lignin, elevating consistently until the 144 hour, and total phenols, maintaining increased levels, highlighted their structural and non-enzymatic contributions to resisting Ascochyta blight.As we traverse beyond conventional resistance mechanisms, the findings unravel opportunities for targeted breeding or genetic engineering. By leveraging the natural defense arsenal within genotypes like GLK 10\u0026ndash;40, GLK 20055, and FLIP-04-219C, the quest for resistant chickpea cultivars gains momentum. This research not only contributes to the scientific understanding of host-pathogen interactions but also holds practical implications for global food security\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAngelini R, Bragaloni M, Federico R, Infantino A, and Porta-Pugua A (1993) Involvement of polyamines, diamine oxidase and peroxidase in resistance of chickpea to \u003cem\u003eAscochyta rabiei.\u003c/em\u003e \u003cem\u003eJ Pl Physiol\u003c/em\u003e \u003cstrong\u003e142:\u003c/strong\u003e 704-09.\u003c/li\u003e\n\u003cli\u003eBahr, L., Castelli, M.V., Barolo, M.I., Mostacero, N.R., Tosello, M.E. and López, S.N., 2016. 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Biochemical analysis of plant protection afforded by a nonpathogenic endophytic mutant of Colletotrichum magna. \u003cem\u003ePlant Physiology\u003c/em\u003e, \u003cem\u003e119\u003c/em\u003e(2), pp.795-804.\u003c/li\u003e\n\u003cli\u003eSarwar, N., Zahid, M. H., Ashfaq, S., \u0026amp; Jamil, F. F. (2011). Induced systemic resistance in chickpea against Ascochyta blight by safe chemicals. \u003cem\u003ePakistan Journal of Botany\u003c/em\u003e, \u003cem\u003e43\u003c/em\u003e(2), 1381-1387.\u003c/li\u003e\n\u003cli\u003eSindhu A N I L, Singh R A J E N D E R, Nehra K S and Singal H R (1995) Elicitor-induced metabolic changes in seedlings of chickpea (Cicer arietinum L.) in relation to Ascochyta blight. \u003cem\u003eAnnals of Biology\u003c/em\u003e \u003cstrong\u003e11:\u003c/strong\u003e 173-83.\u003c/li\u003e\n\u003cli\u003eWise, K.A., Bradley, C.A., Markell, S., Pasche, J., Delgado, J.A., Goswami, R.S. and Gudmestad, N.C., 2011. Sensitivity of Ascochyta rabiei populations to prothioconazole and thiabendazole. \u003cem\u003eCrop Protection\u003c/em\u003e, \u003cem\u003e30\u003c/em\u003e(8), pp.1000-1005.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Chickpea, Kabuli, biochemical, enzymes, resistant, defense","lastPublishedDoi":"10.21203/rs.3.rs-3886689/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3886689/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eChickpea (\u003cem\u003eCicer arietinum\u003c/em\u003e L.), the second-largest global pulse crop, plays a crucial role in providing essential minerals and dietary fiber. Ascochyta blight, caused by the necrotrophic pathogen \u003cem\u003eAscochyta rabie\u003c/em\u003ei, poses a substantial threat to chickpea cultivation. Contemporary cultivars often lose resistance to this disease, necessitating improved management strategies. In this study, six Kabuli chickpea genotypes underwent at treated and controlled conditions, identifying GLK 10\u0026ndash;40, GLK 20055, FLIP 09-194C, FLIP 04-219C, and ICCV 55215 resistant and GLK 17301 susceptible to Ascochyta blight. The exploration into the biochemical responses of these genotypes revealed dynamic shifts in enzymatic activities and biochemical components post-fungal infection. PAL enzyme activity witnessed a remarkable 45.5-fold increase at 96 hours post inoculation in the resistant genotype GLK 10\u0026ndash;40, underscoring its essential role in the early defense cascade against Ascochyta blight. TAL and PPO activity peaked at 96 hours post inoculation notably in GLK 20055 and GLK 10\u0026ndash;40 respectively, emphasizing its participation in the initial defense response. POD activity, a crucial element in plant immunity, reached its peak at 96 hours post inoculation, particularly in the resistant line GLK 20055, signifying prolonged defense mechanisms. Lignin content exhibited a consistent increase till 144 hour post inoculation notably in GLK 20055, highlighting its structural contribution to defense against Ascochyta blight. Total phenol content, crucial in resisting microbial infection, showed heightened levels till 144 hour post inoculation in resistant line FLIP 04-219C maintaining sustained high levels. These findings unveil the biochemical intricacies of chickpea defense mechanisms against Ascochyta blight, laying the groundwork for targeted breeding or genetic engineering. The identified genotypes, such as GLK 10\u0026ndash;40, GLK 20055, and FLIP 04-219C, hold promise for developing resilient chickpea cultivars to counter this challenging disease, crucial for ensuring global food security.\u003c/p\u003e","manuscriptTitle":"Defence mechanism through a biochemical pathway in Kabuli chickpea genotypes against Ascochyta blight","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-01-30 13:46:37","doi":"10.21203/rs.3.rs-3886689/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"94f82196-95fa-4e0c-90af-e7cd69bbc07c","owner":[],"postedDate":"January 30th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-09-19T20:52:50+00:00","versionOfRecord":[],"versionCreatedAt":"2024-01-30 13:46:37","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3886689","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3886689","identity":"rs-3886689","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}