Physiology, Epidemiology and Fungicidal Subdual Strategies for whip smut of sugarcane Caused by Sporisoriums Scitamineum

Physiology, Epidemiology and Fungicidal Subdual Strategies for whip smut of sugarcane Caused by Sporisoriums Scitamineum | 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 Physiology, Epidemiology and Fungicidal Subdual Strategies for whip smut of sugarcane Caused by Sporisoriums Scitamineum Rab Nawaz Noor, Muhammad Atiq, Muhammad Usman, Ameer Jan, Ahmad Nawaz, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7279829/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 Sugarcane, a cornerstone of Pakistan's agricultural economy, standing as the second-largest cash crop in the country. The pervasive threat of whip smut, caused by Sporisorium scitamineum , poses a formidable challenge, often resulting in yield losses ranging from 18–68%. This study explores the incidence of whip smut in sugarcane across Faisalabad, investigating its correlation with environmental factors such as maximum and minimum temperatures (°C), relative humidity (%), rainfall (mm), and wind speed (Km/h), disease impact on plant physiology and effective management strategies using synthetic chemicals. Field experiments, incorporating artificial inoculation across ten sugarcane varieties, revealed varying susceptibility, with HSF-242 exhibiting the highest disease incidence (92%) and CP43-33 the lowest (78%). Statistical analyses highlighted significant negative correlations between disease incidence and maximum/minimum temperatures, while positive correlations were observed with relative humidity, rainfall, and wind speed. In vitro evaluations of ten chemical fungicides identified Aliette (30%), Tilt (40%), and Amistar Top (44.67%) as the most effective in suppressing S. scitamineum . Subsequent field trials confirmed Aliette's superior efficacy, demonstrating its potential in disease management strategies. Photosynthetic rate, transpiration rate, stomatal conductance, and chlorophyll contents (total, a, and b) were observed high in healthy plants as compared to inoculated ones. While water use efficiency was observed in greater amounts in inoculated plants as compared to healthy plants. This study contributes novel insights into managing sugarcane whip smut, offering practical strategies to mitigate economic losses and sustainably enhance crop yields in global sugarcane production systems. Saccharum officinarum Leaf Gasses Exchange Smut Management Epidemiology Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 1. Introduction Sugarcane ( Saccharum officinarum L.), is a tall perennial grass and a vital member of the Poaceae family, belonging to the genus Saccharum (D’Hont et al. 1998 ), known for its stalk's unique ability to store crystallizable sugar (sucrose). As a strictly domesticated plant, sugarcane is believed to be originated in Papua, New Guinea around 8000 B.C., spreading to South East Asia, India, and the Pacific through human migration and hybridization with wild canes (Sreenivasan et al. 1987 ; Fauconnier 1993 ). Sugarcane is cultivated globally in 121 countries, with significant production in Thailand, Pakistan, Bangladesh, China, Brazil, Colombia, India, Cuba, Mexico, Myanmar, Argentina, the Philippines, South Africa, Australia, and the United States (Qureshi 2004 ; Atiq et al. 2023 ). This crop holds considerable economic importance. In 2022, global sugarcane harvest covered 26,089.988 thousand hectares, yielding a total production of 1.92 million metric tons. In Pakistan, sugarcane ranks as the fourth largest and second most important cash crop after cotton, contributing up to 3.5% of agriculture's value added and 0.8% to the GDP. Unambiguously, sugarcane cultivation in Pakistan in 2022 spanned 1,318 thousand hectares, producing 87.98 million metric tons (API 2023; FAO 2024 ). Nutritionally, sugarcane is rich in antioxidants, essential for a strong immune system, and contains small amounts of vitamins, iron, magnesium, lipids, proteins, and carbohydrates. It is used to produce molasses, jaggery, refined sugar, juice, pharmaceuticals, ethanol, biofuels, and energy (Muhammad 2011 ). The productivity of sugarcane is significantly affected by the combination of various biotic and abiotic factors, including bacterial, viral, fungal, and nematode diseases, as well as weeds and animals. Among these, fungal pathogens are some of the most prevalent and destructive biotic threats to sugarcane. Over 100 fungal species cause diseases in sugarcane worldwide and cause about 18–31% losses in crop production (Subhani et al. 2008 ; Verma et al. 2024 ). Amid all the fungal diseases, whip smut caused by the Sporisorium scitamineum (formerly known as Ustilago scitaminea Syd) poses a severe threat to global sugarcane cultivation and can cause yield losses ranging from 18 to 68%, impacting stalk height, girth, and sugar content, thereby complicating harvesting and processing (Sandhu et al. 1969; Xiupeng et al. 2019 ). Sugarcane whip smut, first identified in Natal, South Africa in 1877, infecting sugarcane through buds or germinating shoots, colonizing meristematic tissues and forming whip-like structures filled with black-brown teliospores (Hoy et al. 1986 ; Ferreira and Comstock 1989 ). These structures emerge from lateral buds or terminal meristems, ranging in size from centimeters to meters and enclosed in a silver-white membrane that deteriorates rapidly, releasing teliospores disseminated by wind (Albert and Schenck 1996 ; Rott et al. 2000 ), water, and agricultural practices. The pathogen also banquets through infected seed pieces, persists in soil and related hosts such as Cyperus dilatatus and Saccharum robustum (Santiago et al. 2012 ). In addition, symptoms include stem galls, excessive tillering, and altered inflorescence, all are detrimental to plant growth and yield (Agnihotri et al. 1990; Trione 1980 ). Favoring warm temperatures (25–30°C) and humidity levels of 65–70%, the disease spreads systemically within sugarcane plants worldwide, disrupting the photosynthetic pathway and suppressing defense gene expression upon infection (Waller 1970 ; Comstock 2000 ; Rajput et al. 2021 ). Furthermore, effective management of whip smut in sugarcane requires an integrated approach that combines cultural practices, resistant varieties, and chemical control methods (Atiq et al. 2024 ). Among these strategies, chemical control has proven pivotal in mitigating the impact of S. scitamineum . Key fungicides like Triademifon, Carbendazim, Mancozeb, and Tebuconazole have been extensively employed due to their effectiveness in suppressing fungal growth and reducing disease severity in sugarcane fields (Rajput et al. 2019 ; Raj et al. 2023 ; Atiq et al. 2022 ). These chemicals act by targeting various stages of the fungal life cycle, including spore germination and mycelial growth, thereby inhibiting the formation of whip-like structures filled with teliospores. By controlling existing infections and preventing the spread of the disease to healthy plants, chemical control plays a crucial role in ensuring the health and productivity of sugarcane crops. Therefore, the present study aimed to observe the prevalence of whip smut in District Faisalabad, to examine the interaction between the disease and environmental factors, and to evaluate the effectiveness of synthetic chemicals in managing whip smut of sugarcane. Determining physiological parameters such as photosynthesis, transpiration, stomatal conductance, and chlorophyll contents is essential for evaluating plant health and performance, helping researchers understand how plants respond to biotic stresses. Water-use efficiency and leaf area can guide breeding programs focused on developing resistant and high-yielding crops by selecting for traits that enhance plant performance under varying conditions. Understanding the importance of the issue, our study explored the biochemical compounds present in both healthy and inoculated sugarcane plants. Identifying these compounds can help researchers develop biochemical markers to pinpoint resistant varieties. Additionally, these findings shed light on how plants resist or succumb to diseases, paving the way for more effective strategies to manage disease and other plant infections. 2. Materials and Methods 2.1 Survey and inoculum collection A comprehensive survey was conducted across the five major sugarcane growing sites in Faisalabad, Punjab, Pakistan, viz. Faisalabad Sadar (31°26'44.1"N 73°10'52.6"E), Samundri (31°03'26.1"N 72°56'23.8"E), Jaranwala (31°18'35.2"N 73°25'21.9"E), Jhumra (31°33'33.9"N 73°11'54.0"E), and Tandlianwala ( 31°01'44.2"N 73°07'14.0"E ) , to assess the incidence of whip smut in sugarcane crops (Fig. 1 ). Each site encompassed surveys of five villages, with data collected from three fields per village using a grid-based sampling approach (Stevens 1997 ). Mature whip-like structures were clipped 10–20 cm below the visible apex and air-dried in separate plastic trays for two weeks to gather sugarcane smut spores (teliospores). Teliospores were extracted from the whips by scraping and filtering through a 1 mm × 1 mm sieve, followed by further sieving by using a 53 µm mesh. Twenty-five grams of sieved teliospores were sealed in cellophane bags and stored at 4ºC. Prior to inoculation, teliospore germination on plain agar plates was confirmed at 90% viability, verified through microscopic examination (Johnson and Koenig 1967 ). 2.2 Preparation of Inoculum To prepare the spore suspensions for inoculation, a sterile procedure was meticulously followed, combining 0.1 g of whip smut spores with 100 mL of sterile distilled water (Nasr 1977 ; Wada and Dangana 2016 ). To ensure even distribution of the spores, a drop of Tween-20, a mild surfactant, was added, and the mixture was thoroughly homogenized. The spore concentration in the suspension was quantified by using a Hemocytometer (Counting chambers Neubauer Bright Line; Hausser Scientific, Horsham, Pa.), and viability was assessed by incubating aliquots on water agar (WA) at 30°C for 12 hours (Dawlance CVT Freezers Model VF-1045). Suspensions demonstrating a germination rate exceeding 90% were deemed viable and used for inoculating the seed sets, thus confirming the infectivity of the spores (Bhuiyan et al. 2013 ). Finally, the spore suspension concentration was adjusted to a 5×10⁶ teliospores/mLby using Hemocytometer (Wada et al. 2016 ). 2.3 Seed setts inoculation and Pot Experiments for Fungicide Efficacy A factorial complete randomized design (CRD) with three replications was used in a study at the University of Agriculture Faisalabad's Research Area to evaluate the effectiveness of different fungicides against whip smut of sugarcane. The experiment included 10 different fungicide treatments and a control group that received no fungicide treatment (Table 1 ). Prior to sowing, sets of susceptible sugarcane variety were inoculated with a spore suspension (5×10⁶ teliospores/mL) of S. scitamineum . After 24 hours of inoculation, the sets were treated with the 10 different fungicides for 30 minutes, following the manufacturer's recommended dosage, where an untreated set served as the control. Table 1 List of fungicides used for the chemical control of whip smut of sugarcane caused by Sporisorium scitamineum along with active constituents. Sr.# Fungicides Active Ingredients Mode of Action References 1 Amistar Top Azoxystrobin + Difenoconazole Inhibit the respiration (Wang et al., 2016 ) 2 Triton Validamycin Inhibitor of trehalase (Li et al., 2012 ) 3 Epoxiconazole Epic 25% SC Epoxiconazole Inhibit the metabolism of fungal cells (Xu et al., 2007 ) 4 Tilt Propiconazole C14 demethylation during the production of ergosterol (Gad & Pham, 2014 ). 5 Consist 500 SC Trifloxystrobin 500GI Inhibit spore and mycelial growth (Keith & Walker, 1992 ) 6 Vibrance Duo Fludioxonil 25/L + Sedaxane 25/L Systemic, antifungal agent (Dal Cortivo et al., 2017) 7 Aliette Fosetyl-Al (Alkyl Phosphonate) 800g/kg Currently un-known (Hillebrand et al., 2019 ) 8 Capnazale Captan 70% + Hexaconazole 5% curative, anti-sporulant, and protective properties (Poojashree et al., 2021 ) 9 Cabriotop Pyraclostrobin + Metiram Inhibit germination of spore (Wood & Fisher, 2017 ; Kanungo & Joshi, 2014 ) 10 Revus Start Pipte 430 WG Mandipropamid Inhibit biosynthesis of cell wall (Blum et al., 2010 ) The fungicide suspensions, prepared by using water at ambient temperature, were applied to the treat the sets which were subsequently planted in pots, each filled with 2.5 kg of a homogeneous soil mixture. The experiment involved 33 pots, each containing a set of sugarcane plants, ensuring a robust setup to evaluate the efficacy of the fungicides against whip smut. Three-budded sets were artificially inoculated by soaking them for 20 minutes in a fresh suspension of smut spores (5×10⁶ teliospores/mL) (Abera 2001 ). To create favorable conditions for disease development, the inoculated sets were incubated overnight in water (Wada 2003 ). The experiment with same conditions was repeated to obtain accuracy in results and robust statistical analysis of the data. 2.4 Field-Based Assessment of Fungicides for Whip Smut Management in Sugarcane: Using an RCBD (Randomized Complete Block Design) with three replications, an experiment was carried out in an open field in accordance with manufacturer recommended dose to assess the effectiveness of fungicides against whip smut in sugarcane. Two methods of inoculation, spray and set inoculation, were employed (Table 1 ). A susceptible sugarcane variety was tested using artificial inoculation under natural field conditions. Each treatment consisted of three rows, each 5m long with 1.0m spacing between rows, totaling 15m per treatment. Forty sets of 3-budded seed setts were planted per treatment, amounting to 120 buds. To simulate natural infection, the sets were first inoculated at sowing and then again at the early mature stage, approximately 150 days after planting, using a spore suspension of (5×10⁶ teliospores/mL). Throughout the experiment, standard agronomic practices were meticulously applied until harvest. 2.5 Effect of environmental condition on whip smut of sugarcane Environmental factors, comprising maximum and minimum air temperature (°C), relative humidity (%), rainfall (mm), and wind speed (km/h) were meticulously examined under natural field conditions. These data were sourced from the Meteorology Cell Department of Agronomy, University of Agriculture Faisalabad. The comprehensive data set was leveraged to conduct variance analysis against whip smut disease incidence. Throughout the progression from disease symptom initiation to the harvesting/crop maturity stage, environmental data were diligently recorded. Subsequently, correlations were established and regression analysis was carried out stepwise to discern the relationship between disease incidence and environmental parameters. This meticulous approach enabled a thorough understanding of the interplay between environmental conditions and the prevalence of whip smut disease. 2.6 Data collection: Percentage Disease Incidence data were collected by systematically counting smutted tillers within the cultivated area. Each tiller was meticulously inspected for visible signs of smut infection, and findings were recorded to ensure data accuracy. Disease severity was assessed based on the proportion of infected tillers relative to the total number inspected, providing a comprehensive measure of disease incidence. Seed setts germination data were recorded 45 days after planting, with smut incidence monitored fortnightly until harvest. Any observed smut clumps or whips were promptly removed and destroyed to prevent secondary infestation. Cumulative smut incidence per replicate was calculated based on total germinated seed setts, and growth parameters such as tillers per plant, girth (mm), plant height (cm), mill-able cane per hour, and yield (tons/ha) were measured. Germination percentage and disease incidence were computed by using the following formula: $$\:Incidence\:\left(\%\right)=\frac{\left(\text{N}\text{u}\text{m}\text{b}\text{e}\text{r}\:\text{o}\text{f}\:\text{i}\text{n}\text{f}\text{e}\text{c}\text{t}\text{e}\text{d}\:\text{p}\text{l}\text{a}\text{n}\text{t}\text{s}\right)\text{}}{\text{T}\text{o}\text{t}\text{a}\text{l}\:\text{n}\text{u}\text{m}\text{b}\text{e}\text{r}\:\text{o}\text{f}\:\text{p}\text{l}\text{a}\text{n}\text{t}\text{s}}\times\:100$$ $$\:Germination\:\left(\%\right)=\frac{\left(\text{N}\text{u}\text{m}\text{b}\text{e}\text{r}\:\text{o}\text{f}\:\text{b}\text{u}\text{d}\text{s}\:\text{g}\text{e}\text{r}\text{m}\text{i}\text{n}\text{a}\text{t}\text{e}\text{d}\right)\text{}}{\text{T}\text{o}\text{t}\text{a}\text{l}\:\text{n}\text{u}\text{m}\text{b}\text{e}\text{r}\:\text{o}\text{f}\:\text{b}\text{u}\text{d}\text{s}}\times\:100$$ 2.7 Assessment of morphological attributes To check the effects of Smut pathogen on the physiologycal attributes of sugarcane plant, six previously documented resistat and susceptible varieties were taken from Sugarcane Resarch and Development Board, Faisalabad. These varieties were sown under RCBD after giving sett inoculation. To simulate natural infection, the sets were first inoculated at sowing and then again at the early mature stage, approximately 120 days after planting, using a spore suspension of (5×10⁶ teliospores/mL). All agronomic practices were possitively followed to keep the experiment in good condition. The sample collection and data recording was carried out after 180 days of transplantation. During this period (from transplantation to data recording) the plants were kept under strict observation under green house conditions. 2.7.1 Chlorophyll Contents To determine chlorophyll contents, fresh mango leaves were collected, grinded, and extracted in acetone (80% solution). Centrifugation was done at 12,000 rpm for 5 minutes to purify the enzyme extract. The supernatant was separated, and the absorbance was taken by a spectrophotometer (Model) at 645, 663, and 480 nm (Iqbal et al. 2016 ; Haq et al. 2016 ). While the chlorophyll contents were determined by following formulas: $$\:\text{T}\text{o}\text{t}\text{a}\text{l}\:\text{C}\text{h}\text{l}\text{o}\text{r}\text{o}\text{p}\text{h}\text{y}\text{l}\text{l}\:(\text{m}\text{g}/\text{g})=20.20{A}_{663}\:+\:8.02{A}_{645}$$ $$\:\text{C}\text{h}\text{l}\text{o}\text{r}\text{o}\text{p}\text{h}\text{y}\text{l}\text{l}\:a\:(\text{m}\text{g}/\text{g})=12.7{A}_{663}-2.69{A}_{645}$$ $$\:\text{C}\text{h}\text{l}\text{o}\text{r}\text{o}\text{p}\text{h}\text{y}\text{l}\text{l}\:b\:(\text{m}\text{g}/\text{g})=22.9{A}_{645}-4.68{A}_{663}$$ 2.7.2 Leaf Gas Exchange Parameters Leaf gas exchange parameters, such as net photosynthesis, transpiration rate, and stomatal conductance, were measured on three mature leaves per replication between 10 a.m. and 12 p.m. using the LCi-SD Ultra Compact Photosynthesis System (ADC Bio Scientific Ltd., Global House, Hoddesdon, UK). Intrinsic water-use efficiency, indicating the balance between carbon assimilation and water loss, was determined by calculating the ratio of net photosynthetic rate to stomatal conductance and expressed in µmol mol⁻¹ (Ehleringer and Cerling 1995 ). The measurements were conducted under controlled environmental conditions, including a daytime temperature of 34.6°C, a nighttime temperature of 27°C, a relative humidity of 78.9%, and a 12-hour photoperiod. 2.8 Statistical analysis: Statistical analysis involved analysis of variance (ANOVA) to test for significant differences at a 5% probability level of the treatments (Steel et al. 1997 ). Post hoc Tukey’s test was applied, following ANOVA, and results were graphically presented as mean values across replications. Additionally, correlation analysis and regression models were utilized to examine the relationship between disease incidence and various environmental factors. 3. Results 3.1 Survey for the Collection of Whip Smut Inoculum The study on whip smut of sugarcane in district Faisalabad, Pakistan, highlighted the alarming incidence of this destructive fungal disease. A comprehensive survey covered five major sugarcane growing sites in Faisalabad region of Punjab, Pakistan viz. Faisalabad Sadar, Samundri, Jaranwala, Jhumra, and Tandlianwala, encompassing 75 fields across 25 villages. The research employed a robust data collection protocol, sampling five locations within each field and conducting the process twice to ensure accuracy. Whip smut incidence varied significantly across the surveyed sites, ranging from 14.8% in Samundri to 46.66% in FSD Sadar. On an average, whip smut incidence in Faisalabad district was estimated at 30.74%, underscoring its substantial economic and agricultural threat. Specific village-level incidence rates included Jaranwala at 18.8%, Jhumra at 34.13%, and Tandlianwala at 39.33%. Notably, Samundri's village 462GB and Jaranwala's 206RB exhibited lower disease incidences at 9%, while other villages reported incidences exceeding 54% (Fig. 2 ). 3.2 Seed setts inoculation and Pot Trials for Evaluating Fungicidal Efficacy against Whip Smut of Sugarcane Disease incidence (%) was monitored monthly over a three-month period post-treatment, alongside a control group. Aliette exhibited the highest level of control over whip smut in sugarcane, beginning with a minimal disease incidence of 3.67% in the first month and increasing only slightly to 30% by the third month. This demonstrates its superior effectiveness in managing the disease over time. Tilt, while effective initially with a low incidence of 6.33% in the first month, expressed a gradual decline in the performance, leading to a higher incidence of 44.67% in third month. Amistar Top followed a similar pattern, with disease incidence starting at 10.33% and reaching 40% by the end of the study. Vibrance D and Epoxiconazole showed moderate control, with disease incidence rising to 51.33% and 52.67%, respectively, by the third month. Triton, Consist 50, and Capnazale were less effective, with incidences increasing to between 58.33 and 66.67%. Cabriotop and Revus Star were among the least effective treatments, as their incidences sharply increased to 78.33% and 72%, respectively, by the third month. In comparison, the control group experienced the highest disease incidence, escalating to 87.67%, which highlights the aggressive nature of whip smut in the absence of any treatment (Fig. 3 ). Statistical analysis using the LSD (Least Significant Difference) test revealed significant differences among treatments over time. The average disease incidences over the three-month period were calculated as follows: Amistar Top (22.111%), Triton (35.333%), Epoxiconazole (33.778%), Tilt (22.333%), Consist 50 (39.222%), Vibrance D (29.667%), Aliette (15.222%), Capnazale (41.333%), Cabriotop (53.333%), and Revus star (47.222%). Based on the average incidence values and overall performance, Aliette, Amistar Top and Tilt emerged as the most effective treatments for controlling whip smut in sugarcane (Fig. 3 ). Aliette and Amistar Top consistently demonstrated one of the lowest average disease incidences across the study period, suggesting its robust efficacy. Epoxiconazole and Triton also showed competitive performance, maintaining lower average incidences compared to other treatments. These findings underscore the critical role of fungicide selection and timing in developing effective disease management strategies for sugarcane cultivation. 3.3 Field-Based Assessment of Fungicides for Whip Smut Management in Sugarcane Spray method The study evaluated the efficacy of fungicide treatments against whip smut ( Sporisorium scitamineum ) in sugarcane using spray inoculation, revealing significant differences in disease incidence among treatments. In the first month, Amistar Top (T1) exhibited the highest disease incidence (24.33%), followed by Tilt (T2) at 13.67%, while the control (T0) showed the highest incidence (43.67%). By the second month, disease incidences increased with T0 at 72.33%, T1 at 34.33%, and T2 at 29.33%. Aliette (T3) consistently demonstrated the lowest incidences across months: 10% in the first, 23% in the second, and 46.67% in the third month. Time duration significantly influenced disease incidence, with levels increasing progressively. Throughout the study, T0 (Control) consistently recorded the highest disease incidence, followed by T1 and T2, while T3 consistently showed the lowest incidences. These results highlight the varying effectiveness of Amistar Top (T1), Tilt (T2), and Aliette (T3) in controlling whip smut in sugarcane under the study's conditions and overall, Aliette showed the maximum disease control (Fig. 4 ). 3.4 Assessment of fungicide for the incidence of whip smut of sugarcane using set inoculation The study evaluated the impact of fungicide treatments on whip smut ( Sporisorium scitamineum ) incidence in sugarcane using set inoculation, revealing significant effects of treatments, time durations, and their interaction. Analysis of variance indicated a highly significant difference among treatments for disease incidence. The interaction between treatments and time durations also significantly influenced disease incidence, highlighting the combined effect of treatment selection and timing on disease management. In the first month, the control group (T0) exhibited the highest disease incidence (55.67%), followed by T1 (Amister Top) (33%) and T2 (Tilt) (23.33%), while T3 (Aliette) showed the lowest incidence (19.33%). Disease incidences increased in the second month with T0 at 84.67%, T1 at 45.33%, T2 at 39.67%, and T3 at 34%. By the third month, T0 maintained the highest incidence (97.67%), followed by T1 (69%), T2 (60.33%), and T3 (57.67%) (Fig. 5 ). 3.5 Correlation between environmental factors and cultivars with the whip smut disease development in sugarcane varieties Among all the environmental factors, maximum temperature and minimum temperature showed a significant (P ≤ 0.05) negative correlation, while wind speed showed a significant (P ≤ 0.05) positive correlation with disease incidence (%). The relative humidity and rainfall also showed a significant (P ≥ 0.05) positive correlation with the development of whip smut of sugarcane disease. Five environmental factors (maximum temperature, minimum temperature, relative humidity rainfall, and wind speed) were observed for whip smut of sugarcane disease for ten different varieties (HSF-240, CPF-253, HSF-242, CPF72-2086, CP43-33, CPF-243, CP77-400, SPF213, CPF-237 and SPF-245). Maximum and minimum temperature showed a statistically significant negative correlation with all ten cultivars. T while relative humidity and rainfall and wind speed expressed a significantly positive correlation with all cultivars. (Table 2 ). Table 2 Correlation between environmental factors and cultivars with the whip smut disease incidence Varieties Temperature Relative Humidity % Rainfall (mm) Wind Speed (Km/h) Max ( o C) Min ( o C) HSF-240 -0.5873 0.0447 -0.5659 0.0551 0.6956 0.012 0.7364 0.0063 0.7896 0.0023 CPF-253 -0.5883 0.0442 -0.6224 0.0307 0.6531 0.0213 0.6983 0.0116 0.8282 0.0009 HSF-242 -0.5711 0.0525 -0.568 0.054 0.7041 0.0106 0.7487 0.0051 0.7769 0.003 CPF72-2086 -0.5928 0.0422 -0.5703 0.0529 0.6902 0.013 0.7401 0.0059 0.8021 0.0017 CP43-33 -0.5797 0.0482 -0.5731 0.0514 0.6967 0.0118 0.7527 0.0047 0.7571 0.0044 CPF 243 -0.6231 0.0304 -0.5684 0.0538 0.7273 0.0074 0.7708 0.0033 0.7871 0.0024 CP77-400 -0.5754 0.0503 -0.5733 0.0513 0.6647 0.0184 0.7064 0.0102 0.8342 0.0007 SPF-213 -0.6052 0.0371 -0.5683 0.0539 0.7261 0.0075 0.7779 0.0029 0.7419 0.0057 CPF-237 -0.6293 0.0284 -0.6117 0.0345 0.724 0.0078 0.7676 0.0036 0.7743 0.0031 SPF-245 -0.571 0.0525 -0.6206 0.0313 0.6494 0.0223 0.6908 0.0129 0.8462 0.0005 Upper value = Pearson value *=significant Lower value = correlation value at p is ≤ 0.05value ns = Non-significant 3.6 Characterization of Environmental Factors conducive for the development of Whip Smut of Sugarcane on three Varieties The present study investigated the impact of environmental factors, maximum and minimum temperatures, relative humidity, rainfall, and wind speed on the incidence of whip smut disease in three sugarcane cultivars: HSF-240, CP43-33, and SPF-245 out of ten varieties three varieties were selected on the basis of their susceptibility towards whip smut disease of sugarcane. For regression analysis only those susceptible varieties were selected which expressed significant relationship with epidemiological factors like maximum and minimum temperature, relative humidity, rainfall and wind speed. HSF-240 displayed the highest disease incidence at 92%, followed by CP43-33 at 91% and SPF-245 at 90% under conditions of maximum temperature (31.0°C) (Fig. 6 ) and minimum temperature (19.3°C) (Fig. 7 ). The disease incidence increased proportionally with rising relative humidity, ranging from 37.9–68.0%, observed in HSF-240 (10–92%), CP43-33 (10–91%), and SPF-245 (19–90%) (Fig. 8 ). Similarly, maximum disease incidence (92%) was recorded in HSF-240, CP43-33 (91%), and SPF-245 (90%) under conditions of rainfall (6.6 mm) (Fig. 9 ) and wind speed (3.0 km/h) (Fig. 10 ). These findings underscore the differential susceptibility of sugarcane cultivars to whip smut disease under varying environmental conditions, emphasizing the need for tailored disease management strategies based on local environmental factors. 3.7 Physiological Response of Different Sugarcane Genotypes to Smut Infection The leaf gas exchange parameters, namely photosynthetic rate, transpiration rate, stomatal conductance, intrinsic water-use efficiency, and chlorophyll content, were significantly influenced by whip smut inoculation in the six sugarcane genotypes (Table 3 ). Of the tested genotypes, CPF-237 variety proved superior, having the highest photosynthetic and transpiration rate followed by other varieties. The highest stomatal conductance was statistically registered in CPF-243 variety followed by CPF-237, HSF-240 and so on. Among the six sugarcane genotypes, HSF-240 had significantly higher total, “a”, and “b” chlorophyll contents than other genotypes. In addition, the intrinsic water-use efficiency was recorded in the leaves of CP 43 − 33. Smut inoculation tended to reduce the photosynthetic rate, transpiration rate, stomatal consuctance, and leaf chlorophyll (total, “a”, and “b”) content in all resistance and susceptible genotypes statistically; however, water use efficiency expressed the reverse trend in response to fungal inoculation. Following the smut inoculation, the highest reduction in photosynthetic rate was registered in HSF-240 (20.51%), while the lowest reduction in photosynthetic rate was noticed in CP 43 − 33 (8.13%). Similarly, the highest reduction in transpiration rate was observed in CPF-237 (28.20%). The decrease in stomatal conductance in the tested genotypes following Smut inoculation over control ranged from 10.52 to 23.07%, being lowest in SPF-245 and highest in CPF-243. Similarly, maximum chlorophyll (total, a, and b) contents were found in HSF-240 which were 1.87, 1.34, 0.57 mg g − 1 FW respectively while minimum contents were found in HSF-240 0.67, 0.38, 0.25 mg g − 1 FW respectively. Collectively, smut inoculated plants expressed less rate of physiological attributes than that of un-inoculated ones (Table 3 ). Table 3 Impact of Sporisorium scitamineum inoculation on leaf gas exchange and chlorophyll contents of sugarcane genotypes. Treatment Photosynthetic Rate (µmol m − 2 s − 1 ) Transpiration Rate (mol m –2 s –1 ) Stomatal Conductance (mol m –2 s –1 ) Intrinsic Water-Use Efficiency (µmol H 2 O m − 2 s − 1 ) Total Chlorophyll (mg g − 1 FW) Chlorophyll a (mg g − 1 FW) Chlorophyll b (mg g − 1 FW) Inoculated Resistant CPF-237 6.87 a 0.78 a 0.032 b 199.56 b 1.27 b 0.89 b 0.39 b HSF-240 5.05 c 0.50 cd 0.027 c 174.40 c 1.49 a 0.96 a 0.43 a CPF-243 5.69 b 0.66 b 0.038 a 139.24 d 0.99 d 0.61 d 0.29 d Inoculated Susceptible CP 43 − 33 5.27 bc 0.54 c 0.027 c 209.87 a 0.98 d 0.71 c 0.31 cd SPF-245 4.47 e 0.45 d 0.032 b 135.43 d 1.09 b 0.69 c 0.32 c HSF-240 4.77 d 0.38 e 0.026 c 177.01 c 0.67 e 0.38 e 0.25 e Un-Inoculated Resistant CPF-237 7.21 a 1.00 a 0.038 b 179.24 a 1.57 b 1.09 b 0.43 b HSF-240 6.01 c 0.72 c 0.037 b 149.42 c 1.87 a 1.34 a 0.57 a CPF-243 6.36 b 0.88 b 0.050 a 120.25 e 0.98 d 0.71 e 0.34 d Un-Inoculated Susceptible CP 43 − 33 5.79 d 0.65 d 0.032 c 169.40 b 1.19 c 0.81 d 0.39 c SPF-245 5.09 e 0.57 e 0.036 b 127.45 d 1.23 c 0.89 c 0.41 bc HSF-240 6.27 b 0.54 e 0.037 b 147.24 c 0.87 e 0.61 f 0.33 d Values in the column are mean of each variety and reaction type while lowercase letters were significantly different at p ≤ 0.05 by Tukey’s HSD test. 4. Discussion Whip smut is one of the most prevalent and destructive diseases affecting sugarcane in almost all sugarcane growing regions of the worldwide. It possesses a very high destructive potential, causing substantial yield losses in the susceptible sugarcane cultivars. Effective disease management typically begins with the cultural practices, such as the use of resistant varieties (Iqbal et al. 2022 ). However, in the absence of resistant varieties and the non-availability of effective non-chemical measures, reliance on chemical fungicides becomes indispensable to combat destructive plant diseases and to prevent heavy economic losses (Tahir et al. 2023 ; Usman et al. 2024 ). Therefore, the search for effective fungicides is an ongoing process, as resistance can develop in targeted pathogens and new pathotypes may emerge. In regions considered hot spots for whip smut disease, the application of fungicides for seed setts treatment is a common practice. This study evaluated a wide range of fungicides against whip smut on susceptible variety of sugarcane Cultivation of resistant sugarcane varieties is the most effective and environmentally friendly strategy for managing sugarcane smut disease but if disease appeared in the field in epidemic form then farmers have no option except to use synthetic fungicides. So in contemporary study a number of fungicide were evaluated against whip smut of sugarcane to document their effectiveness. Ten fungicides namely Amistar Top, Triton, Epoxiconazole Epic 25% SC, Tilt, Consist 500 SC, Vibrance Duo, Aliette, Capnazale, CabrioTop, and Revus Start Pipte 430 WG were tested under laboratory conditions. Results indicated that Aliette, Tilt, and Amistar Top were particularly effective in disease management. Aliette, containing Fosetyl-Al (Alkyl Phosphonate) 800g/kg as the active ingredient, was the most efficient. It is known to inhibit fungal growth and spore germination, particularly when combined with other active ingredients. Fosetyl-Al is rapidly absorbed through both leaves and roots and exhibits both acropetal and basipetal translocation, enhancing the plant's natural defense mechanisms against fungal pathogens (Silva et al. 2016 ). Results of the existing study are in line with Rajput et al. ( 2019 ), who demonstrated that fungicides such as Bayleton, Bavistan, and Tilt effectively controlled whip smut when applied as hot water dips. These fungicides were highly effective in reducing disease incidence and enhancing sugarcane growth parameters. Treating seed pieces (setts) with fungicides before planting significantly enhances germination and results in healthier crop stands. Fungicides work by binding to β-tubulin polymers in pathogens, inhibiting essential cellular processes like mitosis, meiosis, and cell shape maintenance (Nene and Thapliyal 1982 ; Horst 2012 ). This disruption of fungal development leads to effective disease management. Tilt, containing Propiconazole as its active ingredient, acts as an inducer of a group of CYPs and an inhibitor of CYP51. According to Nesnow ( 2013 ), Propiconazole activates nuclear receptors that lead to the induction of various CYPs. Amistar Top, which contains azoxystrobin and difenoconazole, provides broad-spectrum activity against a range of diseases. Azoxystrobin offers anti-sporulant activity, while difenoconazole is absorbed by the plant and acts during fungal pathogen penetration and haustoria formation. Our results corroborate these mechanisms, as Tilt and Amistar Top significantly reduced disease incidence and improved plant health in the present experiments. Our findings align with Abera et al. ( 2009 ), who revealed that Tilt, Bayfidan, Bayleton, and Vincit were the most effective treatments for whip smut. Sundravadana et al. ( 2011 ) also effectively controlled the disease using Propiconazole (Tilt) and Triademifon (Bayleton). Bharathi ( 2009 ) recommended treating seed setts with triademifon at 0.1% for 4 hours or propiconazole at 0.1% before planting to eradicate sugarcane smut. Bhuiyan et al. ( 2015 ) Understanding of environmental factors is essential for developing effective strategies to mitigate sugarcane smut disease. As climate change is projected to alter temperature and precipitation patterns, there is a growing urgency to adapt agricultural practices to minimize disease risks. By integrating knowledge of these environmental dynamics into disease management strategies, we can enhance the resilience of sugarcane crops against smut and ensure sustainable agricultural production (Yuan et al. 2024 ; Atiq et al. 2022 ) Environmental conditions play a critical role in the development and spread of the pathogen causing smut disease in sugarcane. According to Que et al. ( 2014 ), airborne teliospores are the primary source of infection in sugarcane-growing regions. Variations in disease severity can be largely attributed to environmental factors, including maximum temperature, minimum temperature, relative humidity, wind speed, and rainfall. Our study investigated these factors in relation to varietal reactions across different sugarcane varieties, revealing statistically significant correlations. Both maximum and minimum temperatures showed a significant negative correlation at P ≤ 0.05% level with disease incidence, indicating reduced disease severity incidence as temperatures increase. This observation is consistent with study of Hidayah et al. ( 2024 ) highlighted the critical influence of temperature on teliospore germination and fungal growth. Hidaya et al. reported a notable reduction in disease incidence to 5.7% at 35°C, with peak incidence recorded between 19.3°C and 31.0°C, identifying these temperature ranges as optimal for disease development. Mansoor et al. ( 2016 ) corroborated these findings, emphasizing the role of temperature in influencing smut disease dynamics. Similarly, wind speed demonstrated a significant positive correlation (P < 0.05) with disease incidence, indicating that higher wind speeds enhance the dispersal of smut teliospores and increased disease likelihood in fields. Mansoor et al. ( 2016 ) found maximum disease incidence (92%) occurring at wind speeds ranging 3.0 km/h, further supporting the impact of wind dynamics on smut epidemiology. Relative humidity and rainfall also exhibited significant positive correlations with whip smut development. Higher relative humidity provides optimal conditions for teliospore germination, with disease incidence increasing as humidity levels rise. Sreeramulu et al. (1971) noted maximum spore dispersal at relative humidity levels of 60 to 70%, consistent with our findings. Mansoor et al. ( 2016 ) also found that relative humidity influences disease incidence, underscoring the importance of moist conditions for disease progression. When a pathogen attacks, plants prioritize the biosynthesis of defense-related substances while reducing other (e.g., growth-related) cellular activities (Usman et al. 2025 ). This permits a reduction in physiological parameters until pathogenic growth has been halted (Bolwell and Wojtaszek 1997 ). This could help the plant by depriving biotrophic pathogens of nutrition. Various physiological parameters, such as chlorophyll contents (a, b, and total), photosynthetic rate, transpiration rate, stomatal conductance, and intrinsic water-use efficiency, were studied in six different genotypes (three resistant and three susceptible) of sugarcane after artificial inoculation. The tested genotypes showed a reduction in their activities due to Smut inoculation. However, the level of reduction was genotype-specific. Susceptible genotypes were found to be worse affected due to Smut inoculation, as the highest reduction in total chlorophyll, chl a, and chl b, stomatal conductance, transpiration rate, and photosynthetic activity were registered in these genotypes. The results of the current study agree with the findings of (Guo et al. 2005 ), who found a significant reduction in chlorophyll content in infected plants with the turnip mosaic virus. Probably, declining chlorophyll content per unit area of leaf, stomatal closure, and reduced stomatal closure upon infection are the reasons for a reduced photosynthetic rate (Shad et al. 2023 ; Sayed 2003 ; Berova et al. 2007 ). The photosynthetic rate in diseased plants is reduced due to destruction of green leaf tissues and defoliation of leaves that leads to decrease in photosynthetic area and ultimately reduce photosynthesis (Cheaib and Nabil 2024 ). Similarly, various photosynthetic parameters like maximum photosynthetic rate, light saturation point, carboxylation efficiency, light compensation point, and dark respiration point in resistant sugarcane varieties was high in resistant plants as compared to susceptible varieties inoculated with pathogens (Hu et al. 2018 ). The rate of transpiration in infected sugarcane plants was less due to reduction in maximal stomatal conductance compared to healthy. Fungal pathogens affect stomatal behavior by changing hormonal signaling pathways including abscisic acid (ABA) which controls stomatal opening and closing (Hajji et al. 2009). Susceptible plants at early stage of infection cause opening of stomata due to attack of pathogens that lead to higher transpiration rates while resistant plants contained pathogens through hypersensitive response (HR) and callose deposition which prevent further spread of bacterial pathogens and close their stomata as a defense mechanism which reduce rate of transpiration (Shad et al. 2024 ; Jeyaraj et al. 2023 ). Bacterial pathogens in infected plants manipulate stomata to remain open, resulting in increased stomatal conductance during early-stage infection (Ahmad et al. 2024 ). Smut fungi secrete effector proteins that suppress salicylic acid signaling and prevent closure of stomata (Atiq et al. 2023 ). During later stage of infection stomatal conductance decreased due to necrosis and damaging of leaf tissues which reduce number of functional stomata (Zeng et al. 2010 ). Resistant plants exhibit controlled stomatal conductance in response to bacterial infection and when threat is neutralized, plant reopen its stomata to restore normal photosynthesis and gas exchanges (Underwood et al. 2007 ). Water use efficiency (WUE) in inoculated plants was low due to production of toxins that damage plant cells and disrupt metabolic processes. Smut fungi also secrete enzymes like cellulases and pectinases, which degrade plant cell walls, further reducing photosynthetic efficiency and WUE. 5. Conclusion Among ten tested fungicides Aliette, Tilt and Amistar top expressed promising results towards whip smut of sugarcane while in case of environmental factors maximum and minimum temperature expressed significant negative correlation while relative humidity, wind speed and rainfall displayed significant positive correlation with incidence of whip smut of sugarcane. Maximum whip smut incidence was observed at 31 o C (maximum) and 19.3 o C as minimum temperature, rainfall 6.6 mm, wind speed 3 km/h and 37.8 to 68% relative humidity. Smut inoculation tended to reduce the photosynthetic rate, transpiration rate, stomatal consuctance, and leaf chlorophyll (total, “a”, and “b”) content in all resistance and susceptible genotypes statistically; however, water use efficiency expressed the reverse trend in response to fungal inoculation. Declarations Competing Interests Authors have declared that no competing interests exist. Author Contribution Writing—original draft, Methodology, Investigation, Formal analysis, Data collection, Conceptualization, RNN, AN, MU and MW Writing – review & editing, Supervision, NAR, MA and AR. Writing – review & editing, Visualization, validation, Formal analysis and references, MUQ, HA and SI. All authors reviewed the manuscript and agreed for publication. 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Biotechnol J 19: e202400457. https://doi.org/10.1002/biot.202400457 Zeng W, Melotto M, He SY (2010) Plant stomata: a checkpoint of host immunity and pathogen virulence. Current Opinions Biotechnol 21(5): 599–603. Additional Declarations No competing interests reported. Supplementary Files GA.png Graphical Abstract 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7279829","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":507209688,"identity":"831a0572-afab-40b7-8ff5-219584dd8049","order_by":0,"name":"Rab Nawaz Noor","email":"","orcid":"","institution":"University of Agriculture","correspondingAuthor":false,"prefix":"","firstName":"Rab","middleName":"Nawaz","lastName":"Noor","suffix":""},{"id":507209689,"identity":"b51d355f-125f-4cfa-a6bb-431144dc6964","order_by":1,"name":"Muhammad Atiq","email":"","orcid":"","institution":"University of Agriculture","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Atiq","suffix":""},{"id":507209690,"identity":"5d1c99b6-74cd-4322-b678-a73b3166f5da","order_by":2,"name":"Muhammad Usman","email":"","orcid":"","institution":"University of Agriculture","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Usman","suffix":""},{"id":507209691,"identity":"290f136b-ef1a-4695-8814-0866dd9860b0","order_by":3,"name":"Ameer Jan","email":"","orcid":"","institution":"University of Agriculture","correspondingAuthor":false,"prefix":"","firstName":"Ameer","middleName":"","lastName":"Jan","suffix":""},{"id":507209692,"identity":"e680b52c-9a5d-4fb7-8a44-9349485818f6","order_by":4,"name":"Ahmad Nawaz","email":"","orcid":"","institution":"University of Agriculture","correspondingAuthor":false,"prefix":"","firstName":"Ahmad","middleName":"","lastName":"Nawaz","suffix":""},{"id":507209693,"identity":"13a79762-cc4f-4e2d-aef5-b3b85b86f552","order_by":5,"name":"Shahid Iqbal","email":"","orcid":"","institution":"University of Agriculture","correspondingAuthor":false,"prefix":"","firstName":"Shahid","middleName":"","lastName":"Iqbal","suffix":""},{"id":507209694,"identity":"c849b684-2886-446c-82b7-bf1aa08ea968","order_by":6,"name":"Muhammad Wahab","email":"","orcid":"","institution":"University of Agriculture","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Wahab","suffix":""},{"id":507209695,"identity":"33f2d721-979c-49a9-bf17-b5646e930017","order_by":7,"name":"Hadeed Ahmad","email":"","orcid":"","institution":"University of Agriculture","correspondingAuthor":false,"prefix":"","firstName":"Hadeed","middleName":"","lastName":"Ahmad","suffix":""},{"id":507209696,"identity":"edca44cc-0b6a-410d-af3d-0c8734dd7288","order_by":8,"name":"Muhammad Hussnain Qaisar","email":"","orcid":"","institution":"University of Agriculture","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"Hussnain","lastName":"Qaisar","suffix":""},{"id":507209697,"identity":"250f54af-20c4-4007-ba5d-79835b29af09","order_by":9,"name":"Nasir Ahmed Rajput","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9UlEQVRIiWNgGAWjYJCCA0DMDMSMDxIqQEzmBmK1MDMbPDgDYjIS1gIFzGySD9tADAJazNmPPzxcuceO3Vwi/7BB4rzaaP52oJYfFdtwarHsyTE4eOZZMrPljGTGB4nbjufOOMzYwNhz5jZOLQYHchgONhwA+uNGMrNB4rZjuQ1ALcyMbXi0nH/+AKilHqSFTSJxzrHc+QS13EgwAGo5DNXSUJO7gbCWNyAtx5ktex4bGyQcO5C7EajlIF6/nE9//LHhQHWyOXviw4c/aupy550/fPDBjwrcWmAg2QBCHwaTBwiqBwI7qJY6YhSPglEwCkbBCAMAqMxi/R5qJgEAAAAASUVORK5CYII=","orcid":"","institution":"University of Agriculture","correspondingAuthor":true,"prefix":"","firstName":"Nasir","middleName":"Ahmed","lastName":"Rajput","suffix":""}],"badges":[],"createdAt":"2025-08-02 18:08:10","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7279829/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7279829/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":90517085,"identity":"0ac0fe12-9116-410c-89f8-9929dc172ec2","added_by":"auto","created_at":"2025-09-03 14:40:49","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":541040,"visible":true,"origin":"","legend":"\u003cp\u003eSurveyed areas of Punjab, Pakistan for the sample collection and documentation of the disease incidence of whip smut of sugarcane.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/c517e41c395d8765095866a1.png"},{"id":90517273,"identity":"fcacf23b-547b-4969-bb5f-deb0b248fd33","added_by":"auto","created_at":"2025-09-03 14:48:49","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":87540,"visible":true,"origin":"","legend":"\u003cp\u003eIncidence (%) of whip smut of sugarcane in various sugarcane growing sites of Faisalabad Punjab, Pakistan\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/27cb5ac22edd4efddd39e56b.png"},{"id":90518317,"identity":"2526524b-e3bc-41c1-a742-d1f9e38d0f93","added_by":"auto","created_at":"2025-09-03 14:56:49","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":181001,"visible":true,"origin":"","legend":"\u003cp\u003eDisease incidence (%) of whip smut in sugarcane recorded over three months with one month interval under different fungicidal treatments. Error bars represent standard errors of the mean. Different letters on bars indicate statistically significant differences among treatments at each time point according to post-hoc test (Tukey’s HSD) at \u003cem\u003ep\u003c/em\u003e ≤ 0.05.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/e84ca495a01570df12a90f5d.png"},{"id":90517274,"identity":"f52a2d36-3a75-4139-a2dc-eca5d11a65dd","added_by":"auto","created_at":"2025-09-03 14:48:49","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":144368,"visible":true,"origin":"","legend":"\u003cp\u003eDisease incidence (%) of whip smut in sugarcane recorded over three months with one month interval under different fungicidal treatments under spray inoculation method. Where T\u003csub\u003e1\u003c/sub\u003e= Amistar Top, T\u003csub\u003e2\u003c/sub\u003e= Tilt and T\u003csub\u003e3\u003c/sub\u003e Aliette. Error bars represent standard errors of the mean. Different letters on bars indicate statistically significant differences among treatments at each time point according to post-hoc test (Tukey’s HSD) at \u003cem\u003ep\u003c/em\u003e ≤ 0.05.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/d35e6e66b05e93cd24710562.png"},{"id":90517087,"identity":"4ecb7bc0-b5b9-4dcc-86fc-304d862e949b","added_by":"auto","created_at":"2025-09-03 14:40:49","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":137319,"visible":true,"origin":"","legend":"\u003cp\u003eDisease incidence (%) of whip smut in sugarcane recorded over three months with one month interval under different fungicidal treatments under sett inoculation method. Where T\u003csub\u003e1\u003c/sub\u003e= Amistar Top, T\u003csub\u003e2\u003c/sub\u003e= Tilt and T\u003csub\u003e3\u003c/sub\u003e Aliette. Each bar represents the mean disease incidence for the respective treatment and time point, with vertical lines indicating standard errors. Different letters above the bars denote statistically significant differences among treatments and time points based on post-hoc test (e.g., LSD or Tukey’s HSD) at \u003cem\u003ep\u003c/em\u003e ≤ 0.05.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/36afb633060b46a83d4b7bf2.png"},{"id":90518578,"identity":"de356537-171d-4c22-ae26-92f037ac6a3b","added_by":"auto","created_at":"2025-09-03 15:04:49","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":101331,"visible":true,"origin":"","legend":"\u003cp\u003eThe relationship between maximum temperature (°C) and disease incidence (%) of whip smut in three sugarcane varieties (HSF-240, CP43-33, and SPF-245). Each data point represents observed disease incidence at a given temperature for the respective variety. Linear regression equations and Pearson correlation coefficients (r) are shown for each variety, indicating a negative correlation between maximum temperature and disease incidence. The results suggest that higher maximum temperatures are associated with reduced incidence of whip smut across all tested sugarcane varieties\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/69f2da6c3ff677a22f8de3ff.png"},{"id":90517276,"identity":"a0ccf68a-fab1-49db-8138-34f5c9597a72","added_by":"auto","created_at":"2025-09-03 14:48:49","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":123255,"visible":true,"origin":"","legend":"\u003cp\u003eThe relationship between minimum temperature (°C) and disease incidence (%) of whip smut in three sugarcane. The regression equations and Pearson correlation coefficients (r) indicate a strong negative correlation between minimum temperature and disease incidence: HSF-240 (y = 79.5714 – 1.7339x, r = –0.5659), CP43-33 (y = 78.4034 – 1.8130x, r = –0.5731), and SPF-245 (y = 88.0664 – 1.9460x, r = –0.6206) suggesting that increasing minimum temperatures are associated with a reduction in whip smut incidence across all tested sugarcane varieties\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/19d200ee0095aeac88e28988.png"},{"id":90517093,"identity":"9c57e686-989e-4bad-8d2b-b6b4afd3920d","added_by":"auto","created_at":"2025-09-03 14:40:49","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":106315,"visible":true,"origin":"","legend":"\u003cp\u003eRelationship between relative humidity (%) and disease incidence (%) of whip smut in three sugarcane varieties: HSF-240, CP43-33, and SPF-245. Disease incidence increased linearly with rising relative humidity across all varieties. Regression equations and correlation coefficients (r) are provided for each variety. Among the tested varieties, CP 43-33 exhibited the highest correlation with relative humidity, suggesting a strong positive influence of wind on disease development.\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/b8f5478097b143afd6a4a585.png"},{"id":90517278,"identity":"c3954623-2c2e-42cd-8308-f2652aa1fcd7","added_by":"auto","created_at":"2025-09-03 14:48:49","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":112635,"visible":true,"origin":"","legend":"\u003cp\u003eRelationship between rainfall (mm) and disease incidence (%) of whip smut in three sugarcane varieties: HSF-240, CP43-33, and SPF-245. Disease incidence increased linearly with rising rainfall across all varieties. Regression equations and correlation coefficients (r) are provided for each variety. Among the tested varieties, CP 43-33 exhibited the highest correlation with rainfall, suggesting a strong positive influence of wind on disease development.\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/830876a6716b4fa72d2db268.png"},{"id":90517110,"identity":"15ea72b5-414d-4056-88c9-5b30885622fd","added_by":"auto","created_at":"2025-09-03 14:40:49","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":121959,"visible":true,"origin":"","legend":"\u003cp\u003eRelationship between wind speed (km/h) and disease incidence (%) of whip smut in three sugarcane varieties: HSF-240, CP43-33, and SPF-245. Disease incidence increased linearly with rising wind speed across all varieties. Regression equations and correlation coefficients (r) are provided for each variety. Among the tested varieties, SPF-245 exhibited the highest correlation with wind speed, suggesting a strong positive influence of wind on disease development.\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/752cb133870dfc196adc5e87.png"},{"id":92512542,"identity":"c582c8de-9138-4fdd-b5d0-f64200ecfc10","added_by":"auto","created_at":"2025-09-30 13:38:58","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2997946,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/113e4f72-198c-44a1-bab2-48acd27ffe9f.pdf"},{"id":90517089,"identity":"d31f22b8-5308-4e85-a46b-4bee3acdf76a","added_by":"auto","created_at":"2025-09-03 14:40:49","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1762160,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical Abstract\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"GA.png","url":"https://assets-eu.researchsquare.com/files/rs-7279829/v1/3028ac9c70c627b511768235.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003ePhysiology, Epidemiology and Fungicidal Subdual Strategies for whip smut of sugarcane Caused by \u003cem\u003eSporisoriums Scitamineum\u003c/em\u003e\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eSugarcane (\u003cem\u003eSaccharum officinarum\u003c/em\u003e L.), is a tall perennial grass and a vital member of the \u003cem\u003ePoaceae\u003c/em\u003e family, belonging to the genus \u003cem\u003eSaccharum\u003c/em\u003e (D\u0026rsquo;Hont et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e1998\u003c/span\u003e), known for its stalk's unique ability to store crystallizable sugar (sucrose). As a strictly domesticated plant, sugarcane is believed to be originated in Papua, New Guinea around 8000 B.C., spreading to South East Asia, India, and the Pacific through human migration and hybridization with wild canes (Sreenivasan et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e1987\u003c/span\u003e; Fauconnier \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e1993\u003c/span\u003e). Sugarcane is cultivated globally in 121 countries, with significant production in Thailand, Pakistan, Bangladesh, China, Brazil, Colombia, India, Cuba, Mexico, Myanmar, Argentina, the Philippines, South Africa, Australia, and the United States (Qureshi \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Atiq et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). This crop holds considerable economic importance. In 2022, global sugarcane harvest covered 26,089.988 thousand hectares, yielding a total production of 1.92\u0026nbsp;million metric tons. In Pakistan, sugarcane ranks as the fourth largest and second most important cash crop after cotton, contributing up to 3.5% of agriculture's value added and 0.8% to the GDP. Unambiguously, sugarcane cultivation in Pakistan in 2022 spanned 1,318 thousand hectares, producing 87.98\u0026nbsp;million metric tons (API 2023; FAO \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Nutritionally, sugarcane is rich in antioxidants, essential for a strong immune system, and contains small amounts of vitamins, iron, magnesium, lipids, proteins, and carbohydrates. It is used to produce molasses, jaggery, refined sugar, juice, pharmaceuticals, ethanol, biofuels, and energy (Muhammad \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe productivity of sugarcane is significantly affected by the combination of various biotic and abiotic factors, including bacterial, viral, fungal, and nematode diseases, as well as weeds and animals. Among these, fungal pathogens are some of the most prevalent and destructive biotic threats to sugarcane. Over 100 fungal species cause diseases in sugarcane worldwide and cause about 18\u0026ndash;31% losses in crop production (Subhani et al. \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Verma et al. \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Amid all the fungal diseases, whip smut caused by the \u003cem\u003eSporisorium scitamineum\u003c/em\u003e (formerly known as \u003cem\u003eUstilago scitaminea\u003c/em\u003e Syd) poses a severe threat to global sugarcane cultivation and can cause yield losses ranging from 18 to 68%, impacting stalk height, girth, and sugar content, thereby complicating harvesting and processing (Sandhu et al. 1969; Xiupeng et al. \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSugarcane whip smut, first identified in Natal, South Africa in 1877, infecting sugarcane through buds or germinating shoots, colonizing meristematic tissues and forming whip-like structures filled with black-brown teliospores (Hoy et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e1986\u003c/span\u003e; Ferreira and Comstock \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e1989\u003c/span\u003e). These structures emerge from lateral buds or terminal meristems, ranging in size from centimeters to meters and enclosed in a silver-white membrane that deteriorates rapidly, releasing teliospores disseminated by wind (Albert and Schenck \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e1996\u003c/span\u003e; Rott et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2000\u003c/span\u003e), water, and agricultural practices. The pathogen also banquets through infected seed pieces, persists in soil and related hosts such as \u003cem\u003eCyperus dilatatus\u003c/em\u003e and \u003cem\u003eSaccharum robustum\u003c/em\u003e (Santiago et al. \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). In addition, symptoms include stem galls, excessive tillering, and altered inflorescence, all are detrimental to plant growth and yield (Agnihotri et al. 1990; Trione \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e1980\u003c/span\u003e). Favoring warm temperatures (25\u0026ndash;30\u0026deg;C) and humidity levels of 65\u0026ndash;70%, the disease spreads systemically within sugarcane plants worldwide, disrupting the photosynthetic pathway and suppressing defense gene expression upon infection (Waller \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e1970\u003c/span\u003e; Comstock \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Rajput et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eFurthermore, effective management of whip smut in sugarcane requires an integrated approach that combines cultural practices, resistant varieties, and chemical control methods (Atiq et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Among these strategies, chemical control has proven pivotal in mitigating the impact of \u003cem\u003eS. scitamineum\u003c/em\u003e. Key fungicides like Triademifon, Carbendazim, Mancozeb, and Tebuconazole have been extensively employed due to their effectiveness in suppressing fungal growth and reducing disease severity in sugarcane fields (Rajput et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Raj et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Atiq et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). These chemicals act by targeting various stages of the fungal life cycle, including spore germination and mycelial growth, thereby inhibiting the formation of whip-like structures filled with teliospores. By controlling existing infections and preventing the spread of the disease to healthy plants, chemical control plays a crucial role in ensuring the health and productivity of sugarcane crops. Therefore, the present study aimed to observe the prevalence of whip smut in District Faisalabad, to examine the interaction between the disease and environmental factors, and to evaluate the effectiveness of synthetic chemicals in managing whip smut of sugarcane. Determining physiological parameters such as photosynthesis, transpiration, stomatal conductance, and chlorophyll contents is essential for evaluating plant health and performance, helping researchers understand how plants respond to biotic stresses. Water-use efficiency and leaf area can guide breeding programs focused on developing resistant and high-yielding crops by selecting for traits that enhance plant performance under varying conditions. Understanding the importance of the issue, our study explored the biochemical compounds present in both healthy and inoculated sugarcane plants. Identifying these compounds can help researchers develop biochemical markers to pinpoint resistant varieties. Additionally, these findings shed light on how plants resist or succumb to diseases, paving the way for more effective strategies to manage disease and other plant infections.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n\u003ch2\u003e2.1 Survey and inoculum collection\u003c/h2\u003e\n\u003cp\u003eA comprehensive survey was conducted across the five major sugarcane growing sites in Faisalabad, Punjab, Pakistan, viz. Faisalabad Sadar (31\u0026deg;26'44.1\"N 73\u0026deg;10'52.6\"E), Samundri (31\u0026deg;03'26.1\"N 72\u0026deg;56'23.8\"E), Jaranwala (31\u0026deg;18'35.2\"N 73\u0026deg;25'21.9\"E), Jhumra (31\u0026deg;33'33.9\"N 73\u0026deg;11'54.0\"E), and Tandlianwala \u003cstrong\u003e(\u003c/strong\u003e31\u0026deg;01'44.2\"N 73\u0026deg;07'14.0\"E\u003cstrong\u003e)\u003c/strong\u003e, to assess the incidence of whip smut in sugarcane crops (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Each site encompassed surveys of five villages, with data collected from three fields per village using a grid-based sampling approach (Stevens \u003cspan class=\"CitationRef\"\u003e1997\u003c/span\u003e). Mature whip-like structures were clipped 10\u0026ndash;20 cm below the visible apex and air-dried in separate plastic trays for two weeks to gather sugarcane smut spores (teliospores). Teliospores were extracted from the whips by scraping and filtering through a 1 mm \u0026times; 1 mm sieve, followed by further sieving by using a 53 \u0026micro;m mesh. Twenty-five grams of sieved teliospores were sealed in cellophane bags and stored at 4\u0026ordm;C. Prior to inoculation, teliospore germination on plain agar plates was confirmed at 90% viability, verified through microscopic examination (Johnson and Koenig \u003cspan class=\"CitationRef\"\u003e1967\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n\u003ch2\u003e2.2 Preparation of Inoculum\u003c/h2\u003e\n\u003cp\u003eTo prepare the spore suspensions for inoculation, a sterile procedure was meticulously followed, combining 0.1 g of whip smut spores with 100 mL of sterile distilled water (Nasr \u003cspan class=\"CitationRef\"\u003e1977\u003c/span\u003e; Wada and Dangana \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e). To ensure even distribution of the spores, a drop of Tween-20, a mild surfactant, was added, and the mixture was thoroughly homogenized. The spore concentration in the suspension was quantified by using a Hemocytometer (Counting chambers Neubauer Bright Line; Hausser Scientific, Horsham, Pa.), and viability was assessed by incubating aliquots on water agar (WA) at 30\u0026deg;C for 12 hours (Dawlance CVT Freezers Model VF-1045). Suspensions demonstrating a germination rate exceeding 90% were deemed viable and used for inoculating the seed sets, thus confirming the infectivity of the spores (Bhuiyan et al. \u003cspan class=\"CitationRef\"\u003e2013\u003c/span\u003e). Finally, the spore suspension concentration was adjusted to a 5\u0026times;10⁶ teliospores/mLby using Hemocytometer (Wada et al. \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n\u003ch2\u003e2.3 Seed setts inoculation and Pot Experiments for Fungicide Efficacy\u003c/h2\u003e\n\u003cp\u003eA factorial complete randomized design (CRD) with three replications was used in a study at the University of Agriculture Faisalabad's Research Area to evaluate the effectiveness of different fungicides against whip smut of sugarcane. The experiment included 10 different fungicide treatments and a control group that received no fungicide treatment (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Prior to sowing, sets of susceptible sugarcane variety were inoculated with a spore suspension (5\u0026times;10⁶ teliospores/mL) of \u003cem\u003eS. scitamineum\u003c/em\u003e. After 24 hours of inoculation, the sets were treated with the 10 different fungicides for 30 minutes, following the manufacturer's recommended dosage, where an untreated set served as the control.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eList of fungicides used for the chemical control of whip smut of sugarcane caused by \u003cem\u003eSporisorium scitamineum\u003c/em\u003e along with active constituents.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSr.#\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eFungicides\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eActive Ingredients\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMode of Action\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eReferences\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAmistar Top\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAzoxystrobin + Difenoconazole\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eInhibit the respiration\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(Wang et al., \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTriton\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eValidamycin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eInhibitor of trehalase\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(Li et al., \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eEpoxiconazole Epic 25% SC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eEpoxiconazole\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eInhibit the metabolism of fungal cells\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(Xu et al., \u003cspan class=\"CitationRef\"\u003e2007\u003c/span\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTilt\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePropiconazole\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC14 demethylation during the production of ergosterol\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(Gad \u0026amp; Pham, \u003cspan class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eConsist 500 SC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTrifloxystrobin 500GI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eInhibit spore and mycelial growth\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(Keith \u0026amp; Walker, \u003cspan class=\"CitationRef\"\u003e1992\u003c/span\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eVibrance Duo\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFludioxonil 25/L\u0026thinsp;+\u0026thinsp;Sedaxane 25/L\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSystemic, antifungal agent\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(Dal Cortivo et al., 2017)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAliette\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFosetyl-Al (Alkyl Phosphonate) 800g/kg\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCurrently un-known\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(Hillebrand et al., \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCapnazale\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCaptan 70% + Hexaconazole 5%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ecurative, anti-sporulant, and protective properties\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(Poojashree et al., \u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCabriotop\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePyraclostrobin\u0026thinsp;+\u0026thinsp;Metiram\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eInhibit germination of spore\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(Wood \u0026amp; Fisher, \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e; Kanungo \u0026amp; Joshi, \u003cspan class=\"CitationRef\"\u003e2014\u003c/span\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eRevus Start Pipte 430 WG\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eMandipropamid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eInhibit biosynthesis of cell wall\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(Blum et al., \u003cspan class=\"CitationRef\"\u003e2010\u003c/span\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eThe fungicide suspensions, prepared by using water at ambient temperature, were applied to the treat the sets which were subsequently planted in pots, each filled with 2.5 kg of a homogeneous soil mixture. The experiment involved 33 pots, each containing a set of sugarcane plants, ensuring a robust setup to evaluate the efficacy of the fungicides against whip smut. Three-budded sets were artificially inoculated by soaking them for 20 minutes in a fresh suspension of smut spores (5\u0026times;10⁶ teliospores/mL) (Abera \u003cspan class=\"CitationRef\"\u003e2001\u003c/span\u003e). To create favorable conditions for disease development, the inoculated sets were incubated overnight in water (Wada \u003cspan class=\"CitationRef\"\u003e2003\u003c/span\u003e). The experiment with same conditions was repeated to obtain accuracy in results and robust statistical analysis of the data.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n\u003ch2\u003e2.4 Field-Based Assessment of Fungicides for Whip Smut Management in Sugarcane:\u003c/h2\u003e\n\u003cp\u003eUsing an RCBD (Randomized Complete Block Design) with three replications, an experiment was carried out in an open field in accordance with manufacturer recommended dose to assess the effectiveness of fungicides against whip smut in sugarcane. Two methods of inoculation, spray and set inoculation, were employed (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). A susceptible sugarcane variety was tested using artificial inoculation under natural field conditions. Each treatment consisted of three rows, each 5m long with 1.0m spacing between rows, totaling 15m per treatment. Forty sets of 3-budded seed setts were planted per treatment, amounting to 120 buds. To simulate natural infection, the sets were first inoculated at sowing and then again at the early mature stage, approximately 150 days after planting, using a spore suspension of (5\u0026times;10⁶ teliospores/mL). Throughout the experiment, standard agronomic practices were meticulously applied until harvest.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n\u003ch2\u003e2.5 Effect of environmental condition on whip smut of sugarcane\u003c/h2\u003e\n\u003cp\u003eEnvironmental factors, comprising maximum and minimum air temperature (\u0026deg;C), relative humidity (%), rainfall (mm), and wind speed (km/h) were meticulously examined under natural field conditions. These data were sourced from the Meteorology Cell Department of Agronomy, University of Agriculture Faisalabad. The comprehensive data set was leveraged to conduct variance analysis against whip smut disease incidence. Throughout the progression from disease symptom initiation to the harvesting/crop maturity stage, environmental data were diligently recorded. Subsequently, correlations were established and regression analysis was carried out stepwise to discern the relationship between disease incidence and environmental parameters. This meticulous approach enabled a thorough understanding of the interplay between environmental conditions and the prevalence of whip smut disease.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n\u003ch2\u003e2.6 Data collection:\u003c/h2\u003e\n\u003cp\u003ePercentage Disease Incidence data were collected by systematically counting smutted tillers within the cultivated area. Each tiller was meticulously inspected for visible signs of smut infection, and findings were recorded to ensure data accuracy. Disease severity was assessed based on the proportion of infected tillers relative to the total number inspected, providing a comprehensive measure of disease incidence. Seed setts germination data were recorded 45 days after planting, with smut incidence monitored fortnightly until harvest. Any observed smut clumps or whips were promptly removed and destroyed to prevent secondary infestation. Cumulative smut incidence per replicate was calculated based on total germinated seed setts, and growth parameters such as tillers per plant, girth (mm), plant height (cm), mill-able cane per hour, and yield (tons/ha) were measured. Germination percentage and disease incidence were computed by using the following formula:\u003c/p\u003e\n\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\n\u003cdiv id=\"FileID_Equa\" class=\"mathdisplay\"\u003e$$\\:Incidence\\:\\left(\\%\\right)=\\frac{\\left(\\text{N}\\text{u}\\text{m}\\text{b}\\text{e}\\text{r}\\:\\text{o}\\text{f}\\:\\text{i}\\text{n}\\text{f}\\text{e}\\text{c}\\text{t}\\text{e}\\text{d}\\:\\text{p}\\text{l}\\text{a}\\text{n}\\text{t}\\text{s}\\right)\\text{}}{\\text{T}\\text{o}\\text{t}\\text{a}\\text{l}\\:\\text{n}\\text{u}\\text{m}\\text{b}\\text{e}\\text{r}\\:\\text{o}\\text{f}\\:\\text{p}\\text{l}\\text{a}\\text{n}\\text{t}\\text{s}}\\times\\:100$$\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\n\u003cdiv id=\"FileID_Equb\" class=\"mathdisplay\"\u003e$$\\:Germination\\:\\left(\\%\\right)=\\frac{\\left(\\text{N}\\text{u}\\text{m}\\text{b}\\text{e}\\text{r}\\:\\text{o}\\text{f}\\:\\text{b}\\text{u}\\text{d}\\text{s}\\:\\text{g}\\text{e}\\text{r}\\text{m}\\text{i}\\text{n}\\text{a}\\text{t}\\text{e}\\text{d}\\right)\\text{}}{\\text{T}\\text{o}\\text{t}\\text{a}\\text{l}\\:\\text{n}\\text{u}\\text{m}\\text{b}\\text{e}\\text{r}\\:\\text{o}\\text{f}\\:\\text{b}\\text{u}\\text{d}\\text{s}}\\times\\:100$$\u003c/div\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n\u003ch2\u003e2.7 Assessment of morphological attributes\u003c/h2\u003e\n\u003cp\u003eTo check the effects of Smut pathogen on the physiologycal attributes of sugarcane plant, six previously documented resistat and susceptible varieties were taken from Sugarcane Resarch and Development Board, Faisalabad. These varieties were sown under RCBD after giving sett inoculation. To simulate natural infection, the sets were first inoculated at sowing and then again at the early mature stage, approximately 120 days after planting, using a spore suspension of (5\u0026times;10⁶ teliospores/mL). All agronomic practices were possitively followed to keep the experiment in good condition. The sample collection and data recording was carried out after 180 days of transplantation. During this period (from transplantation to data recording) the plants were kept under strict observation under green house conditions.\u003c/p\u003e\n\u003cdiv id=\"Sec10\" class=\"Section3\"\u003e\n\u003ch2\u003e2.7.1 Chlorophyll Contents\u003c/h2\u003e\n\u003cp\u003eTo determine chlorophyll contents, fresh mango leaves were collected, grinded, and extracted in acetone (80% solution). Centrifugation was done at 12,000 rpm for 5 minutes to purify the enzyme extract. The supernatant was separated, and the absorbance was taken by a spectrophotometer (Model) at 645, 663, and 480 nm (Iqbal et al. \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e; Haq et al. \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e). While the chlorophyll contents were determined by following formulas:\u003c/p\u003e\n\u003cdiv id=\"Equc\" class=\"Equation\"\u003e\n\u003cdiv id=\"FileID_Equc\" class=\"mathdisplay\"\u003e$$\\:\\text{T}\\text{o}\\text{t}\\text{a}\\text{l}\\:\\text{C}\\text{h}\\text{l}\\text{o}\\text{r}\\text{o}\\text{p}\\text{h}\\text{y}\\text{l}\\text{l}\\:(\\text{m}\\text{g}/\\text{g})=20.20{A}_{663}\\:+\\:8.02{A}_{645}$$\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Equd\" class=\"Equation\"\u003e\n\u003cdiv id=\"FileID_Equd\" class=\"mathdisplay\"\u003e$$\\:\\text{C}\\text{h}\\text{l}\\text{o}\\text{r}\\text{o}\\text{p}\\text{h}\\text{y}\\text{l}\\text{l}\\:a\\:(\\text{m}\\text{g}/\\text{g})=12.7{A}_{663}-2.69{A}_{645}$$\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Eque\" class=\"Equation\"\u003e\n\u003cdiv id=\"FileID_Eque\" class=\"mathdisplay\"\u003e$$\\:\\text{C}\\text{h}\\text{l}\\text{o}\\text{r}\\text{o}\\text{p}\\text{h}\\text{y}\\text{l}\\text{l}\\:b\\:(\\text{m}\\text{g}/\\text{g})=22.9{A}_{645}-4.68{A}_{663}$$\u003c/div\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section3\"\u003e\n\u003ch2\u003e2.7.2 Leaf Gas Exchange Parameters\u003c/h2\u003e\n\u003cp\u003eLeaf gas exchange parameters, such as net photosynthesis, transpiration rate, and stomatal conductance, were measured on three mature leaves per replication between 10 a.m. and 12 p.m. using the LCi-SD Ultra Compact Photosynthesis System (ADC Bio Scientific Ltd., Global House, Hoddesdon, UK). Intrinsic water-use efficiency, indicating the balance between carbon assimilation and water loss, was determined by calculating the ratio of net photosynthetic rate to stomatal conductance and expressed in \u0026micro;mol mol⁻\u0026sup1; (Ehleringer and Cerling \u003cspan class=\"CitationRef\"\u003e1995\u003c/span\u003e). The measurements were conducted under controlled environmental conditions, including a daytime temperature of 34.6\u0026deg;C, a nighttime temperature of 27\u0026deg;C, a relative humidity of 78.9%, and a 12-hour photoperiod.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n\u003ch2\u003e2.8 Statistical analysis:\u003c/h2\u003e\n\u003cp\u003eStatistical analysis involved analysis of variance (ANOVA) to test for significant differences at a 5% probability level of the treatments (Steel et al. \u003cspan class=\"CitationRef\"\u003e1997\u003c/span\u003e). Post hoc Tukey\u0026rsquo;s test was applied, following ANOVA, and results were graphically presented as mean values across replications. Additionally, correlation analysis and regression models were utilized to examine the relationship between disease incidence and various environmental factors.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n\u003ch2\u003e3.1 Survey for the Collection of Whip Smut Inoculum\u003c/h2\u003e\n\u003cp\u003eThe study on whip smut of sugarcane in district Faisalabad, Pakistan, highlighted the alarming incidence of this destructive fungal disease. A comprehensive survey covered five major sugarcane growing sites in Faisalabad region of Punjab, Pakistan viz. Faisalabad Sadar, Samundri, Jaranwala, Jhumra, and Tandlianwala, encompassing 75 fields across 25 villages. The research employed a robust data collection protocol, sampling five locations within each field and conducting the process twice to ensure accuracy.\u003c/p\u003e\n\u003cp\u003eWhip smut incidence varied significantly across the surveyed sites, ranging from 14.8% in Samundri to 46.66% in FSD Sadar. On an average, whip smut incidence in Faisalabad district was estimated at 30.74%, underscoring its substantial economic and agricultural threat. Specific village-level incidence rates included Jaranwala at 18.8%, Jhumra at 34.13%, and Tandlianwala at 39.33%. Notably, Samundri's village 462GB and Jaranwala's 206RB exhibited lower disease incidences at 9%, while other villages reported incidences exceeding 54% (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n\u003ch2\u003e3.2 Seed setts inoculation and Pot Trials for Evaluating Fungicidal Efficacy against Whip Smut of Sugarcane\u003c/h2\u003e\n\u003cp\u003eDisease incidence (%) was monitored monthly over a three-month period post-treatment, alongside a control group. Aliette exhibited the highest level of control over whip smut in sugarcane, beginning with a minimal disease incidence of 3.67% in the first month and increasing only slightly to 30% by the third month. This demonstrates its superior effectiveness in managing the disease over time. Tilt, while effective initially with a low incidence of 6.33% in the first month, expressed a gradual decline in the performance, leading to a higher incidence of 44.67% in third month. Amistar Top followed a similar pattern, with disease incidence starting at 10.33% and reaching 40% by the end of the study. Vibrance D and Epoxiconazole showed moderate control, with disease incidence rising to 51.33% and 52.67%, respectively, by the third month. Triton, Consist 50, and Capnazale were less effective, with incidences increasing to between 58.33 and 66.67%. Cabriotop and Revus Star were among the least effective treatments, as their incidences sharply increased to 78.33% and 72%, respectively, by the third month. In comparison, the control group experienced the highest disease incidence, escalating to 87.67%, which highlights the aggressive nature of whip smut in the absence of any treatment (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eStatistical analysis using the LSD (Least Significant Difference) test revealed significant differences among treatments over time. The average disease incidences over the three-month period were calculated as follows: Amistar Top (22.111%), Triton (35.333%), Epoxiconazole (33.778%), Tilt (22.333%), Consist 50 (39.222%), Vibrance D (29.667%), Aliette (15.222%), Capnazale (41.333%), Cabriotop (53.333%), and Revus star (47.222%). Based on the average incidence values and overall performance, Aliette, Amistar Top and Tilt emerged as the most effective treatments for controlling whip smut in sugarcane (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). Aliette and Amistar Top consistently demonstrated one of the lowest average disease incidences across the study period, suggesting its robust efficacy. Epoxiconazole and Triton also showed competitive performance, maintaining lower average incidences compared to other treatments. These findings underscore the critical role of fungicide selection and timing in developing effective disease management strategies for sugarcane cultivation.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n\u003ch2\u003e3.3 Field-Based Assessment of Fungicides for Whip Smut Management in Sugarcane\u003c/h2\u003e\n\u003cp\u003e\u003cstrong\u003eSpray method\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study evaluated the efficacy of fungicide treatments against whip smut (\u003cem\u003eSporisorium scitamineum\u003c/em\u003e) in sugarcane using spray inoculation, revealing significant differences in disease incidence among treatments. In the first month, Amistar Top (T1) exhibited the highest disease incidence (24.33%), followed by Tilt (T2) at 13.67%, while the control (T0) showed the highest incidence (43.67%). By the second month, disease incidences increased with T0 at 72.33%, T1 at 34.33%, and T2 at 29.33%. Aliette (T3) consistently demonstrated the lowest incidences across months: 10% in the first, 23% in the second, and 46.67% in the third month. Time duration significantly influenced disease incidence, with levels increasing progressively. Throughout the study, T0 (Control) consistently recorded the highest disease incidence, followed by T1 and T2, while T3 consistently showed the lowest incidences. These results highlight the varying effectiveness of Amistar Top (T1), Tilt (T2), and Aliette (T3) in controlling whip smut in sugarcane under the study's conditions and overall, Aliette showed the maximum disease control (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n\u003ch2\u003e3.4 Assessment of fungicide for the incidence of whip smut of sugarcane using set inoculation\u003c/h2\u003e\n\u003cp\u003eThe study evaluated the impact of fungicide treatments on whip smut (\u003cem\u003eSporisorium scitamineum\u003c/em\u003e) incidence in sugarcane using set inoculation, revealing significant effects of treatments, time durations, and their interaction. Analysis of variance indicated a highly significant difference among treatments for disease incidence. The interaction between treatments and time durations also significantly influenced disease incidence, highlighting the combined effect of treatment selection and timing on disease management. In the first month, the control group (T0) exhibited the highest disease incidence (55.67%), followed by T1 (Amister Top) (33%) and T2 (Tilt) (23.33%), while T3 (Aliette) showed the lowest incidence (19.33%). Disease incidences increased in the second month with T0 at 84.67%, T1 at 45.33%, T2 at 39.67%, and T3 at 34%. By the third month, T0 maintained the highest incidence (97.67%), followed by T1 (69%), T2 (60.33%), and T3 (57.67%) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n\u003ch2\u003e3.5 Correlation between environmental factors and cultivars with the whip smut disease development in sugarcane varieties\u003c/h2\u003e\n\u003cp\u003eAmong all the environmental factors, maximum temperature and minimum temperature showed a significant (P\u0026thinsp;\u003cspan class=\"Underline\"\u003e\u0026le;\u003c/span\u003e\u0026thinsp;0.05) negative correlation, while wind speed showed a significant (P\u0026thinsp;\u003cspan class=\"Underline\"\u003e\u0026le;\u003c/span\u003e\u0026thinsp;0.05) positive correlation with disease incidence (%). The relative humidity and rainfall also showed a significant (P\u0026thinsp;\u003cspan class=\"Underline\"\u003e\u0026ge;\u003c/span\u003e\u0026thinsp;0.05) positive correlation with the development of whip smut of sugarcane disease. Five environmental factors (maximum temperature, minimum temperature, relative humidity rainfall, and wind speed) were observed for whip smut of sugarcane disease for ten different varieties (HSF-240, CPF-253, HSF-242, CPF72-2086, CP43-33, CPF-243, CP77-400, SPF213, CPF-237 and SPF-245). Maximum and minimum temperature showed a statistically significant negative correlation with all ten cultivars. T while relative humidity and rainfall and wind speed expressed a significantly positive correlation with all cultivars. (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eCorrelation between environmental factors and cultivars with the whip smut disease incidence\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eVarieties\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eTemperature\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eRelative Humidity %\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eRainfall (mm)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eWind Speed (Km/h)\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMax (\u003csup\u003eo\u003c/sup\u003eC)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMin (\u003csup\u003eo\u003c/sup\u003eC)\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eHSF-240\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5873\u003c/p\u003e\n\u003cp\u003e0.0447\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5659\u003c/p\u003e\n\u003cp\u003e0.0551\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6956\u003c/p\u003e\n\u003cp\u003e0.012\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7364\u003c/p\u003e\n\u003cp\u003e0.0063\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7896\u003c/p\u003e\n\u003cp\u003e0.0023\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCPF-253\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5883\u003c/p\u003e\n\u003cp\u003e0.0442\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.6224\u003c/p\u003e\n\u003cp\u003e0.0307\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6531\u003c/p\u003e\n\u003cp\u003e0.0213\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6983\u003c/p\u003e\n\u003cp\u003e0.0116\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8282\u003c/p\u003e\n\u003cp\u003e0.0009\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eHSF-242\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5711\u003c/p\u003e\n\u003cp\u003e0.0525\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.568\u003c/p\u003e\n\u003cp\u003e0.054\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7041\u003c/p\u003e\n\u003cp\u003e0.0106\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7487\u003c/p\u003e\n\u003cp\u003e0.0051\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7769\u003c/p\u003e\n\u003cp\u003e0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCPF72-2086\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5928\u003c/p\u003e\n\u003cp\u003e0.0422\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5703\u003c/p\u003e\n\u003cp\u003e0.0529\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6902\u003c/p\u003e\n\u003cp\u003e0.013\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7401\u003c/p\u003e\n\u003cp\u003e0.0059\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8021\u003c/p\u003e\n\u003cp\u003e0.0017\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCP43-33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5797\u003c/p\u003e\n\u003cp\u003e0.0482\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5731\u003c/p\u003e\n\u003cp\u003e0.0514\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6967\u003c/p\u003e\n\u003cp\u003e0.0118\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7527\u003c/p\u003e\n\u003cp\u003e0.0047\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7571\u003c/p\u003e\n\u003cp\u003e0.0044\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCPF 243\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.6231\u003c/p\u003e\n\u003cp\u003e0.0304\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5684\u003c/p\u003e\n\u003cp\u003e0.0538\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7273\u003c/p\u003e\n\u003cp\u003e0.0074\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7708\u003c/p\u003e\n\u003cp\u003e0.0033\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7871\u003c/p\u003e\n\u003cp\u003e0.0024\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCP77-400\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5754\u003c/p\u003e\n\u003cp\u003e0.0503\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5733\u003c/p\u003e\n\u003cp\u003e0.0513\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6647\u003c/p\u003e\n\u003cp\u003e0.0184\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7064\u003c/p\u003e\n\u003cp\u003e0.0102\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8342\u003c/p\u003e\n\u003cp\u003e0.0007\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSPF-213\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.6052\u003c/p\u003e\n\u003cp\u003e0.0371\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.5683\u003c/p\u003e\n\u003cp\u003e0.0539\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7261\u003c/p\u003e\n\u003cp\u003e0.0075\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7779\u003c/p\u003e\n\u003cp\u003e0.0029\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7419\u003c/p\u003e\n\u003cp\u003e0.0057\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCPF-237\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.6293\u003c/p\u003e\n\u003cp\u003e0.0284\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.6117\u003c/p\u003e\n\u003cp\u003e0.0345\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.724\u003c/p\u003e\n\u003cp\u003e0.0078\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7676\u003c/p\u003e\n\u003cp\u003e0.0036\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7743\u003c/p\u003e\n\u003cp\u003e0.0031\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSPF-245\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.571\u003c/p\u003e\n\u003cp\u003e0.0525\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.6206\u003c/p\u003e\n\u003cp\u003e0.0313\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6494\u003c/p\u003e\n\u003cp\u003e0.0223\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6908\u003c/p\u003e\n\u003cp\u003e0.0129\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8462\u003c/p\u003e\n\u003cp\u003e0.0005\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\"\u003eUpper value\u0026thinsp;=\u0026thinsp;Pearson value *=significant\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\"\u003eLower value\u0026thinsp;=\u0026thinsp;correlation value at p is \u0026le;\u0026thinsp;0.05value ns\u0026thinsp;=\u0026thinsp;Non-significant\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e3.6 Characterization of Environmental Factors conducive for the development of Whip Smut of Sugarcane on three Varieties\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe present study investigated the impact of environmental factors, maximum and minimum temperatures, relative humidity, rainfall, and wind speed on the incidence of whip smut disease in three sugarcane cultivars: HSF-240, CP43-33, and SPF-245 out of ten varieties three varieties were selected on the basis of their susceptibility towards whip smut disease of sugarcane. For regression analysis only those susceptible varieties were selected which expressed significant relationship with epidemiological factors like maximum and minimum temperature, relative humidity, rainfall and wind speed.\u003c/p\u003e\n\u003cp\u003eHSF-240 displayed the highest disease incidence at 92%, followed by CP43-33 at 91% and SPF-245 at 90% under conditions of maximum temperature (31.0\u0026deg;C) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e) and minimum temperature (19.3\u0026deg;C) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e). The disease incidence increased proportionally with rising relative humidity, ranging from 37.9\u0026ndash;68.0%, observed in HSF-240 (10\u0026ndash;92%), CP43-33 (10\u0026ndash;91%), and SPF-245 (19\u0026ndash;90%) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e8\u003c/span\u003e). Similarly, maximum disease incidence (92%) was recorded in HSF-240, CP43-33 (91%), and SPF-245 (90%) under conditions of rainfall (6.6 mm) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e9\u003c/span\u003e) and wind speed (3.0 km/h) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e10\u003c/span\u003e). These findings underscore the differential susceptibility of sugarcane cultivars to whip smut disease under varying environmental conditions, emphasizing the need for tailored disease management strategies based on local environmental factors.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\n\u003ch2\u003e3.7 \u003cstrong\u003ePhysiological Response of Different\u003c/strong\u003e Sugarcane \u003cstrong\u003eGenotypes to Smut Infection\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eThe leaf gas exchange parameters, namely photosynthetic rate, transpiration rate, stomatal conductance, intrinsic water-use efficiency, and chlorophyll content, were significantly influenced by whip smut inoculation in the six sugarcane genotypes (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). Of the tested genotypes, CPF-237 variety proved superior, having the highest photosynthetic and transpiration rate followed by other varieties. The highest stomatal conductance was statistically registered in CPF-243 variety followed by CPF-237, HSF-240 and so on. Among the six sugarcane genotypes, HSF-240 had significantly higher total, \u0026ldquo;a\u0026rdquo;, and \u0026ldquo;b\u0026rdquo; chlorophyll contents than other genotypes. In addition, the intrinsic water-use efficiency was recorded in the leaves of CP 43\u0026thinsp;\u0026minus;\u0026thinsp;33. Smut inoculation tended to reduce the photosynthetic rate, transpiration rate, stomatal consuctance, and leaf chlorophyll (total, \u0026ldquo;a\u0026rdquo;, and \u0026ldquo;b\u0026rdquo;) content in all resistance and susceptible genotypes statistically; however, water use efficiency expressed the reverse trend in response to fungal inoculation. Following the smut inoculation, the highest reduction in photosynthetic rate was registered in HSF-240 (20.51%), while the lowest reduction in photosynthetic rate was noticed in CP 43\u0026thinsp;\u0026minus;\u0026thinsp;33 (8.13%). Similarly, the highest reduction in transpiration rate was observed in CPF-237 (28.20%). The decrease in stomatal conductance in the tested genotypes following Smut inoculation over control ranged from 10.52 to 23.07%, being lowest in SPF-245 and highest in CPF-243. Similarly, maximum chlorophyll (total, a, and b) contents were found in HSF-240 which were 1.87, 1.34, 0.57 mg g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW respectively while minimum contents were found in HSF-240 0.67, 0.38, 0.25 mg g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e FW respectively. Collectively, smut inoculated plants expressed less rate of physiological attributes than that of un-inoculated ones (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab3\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eImpact of \u003cem\u003eSporisorium scitamineum\u003c/em\u003e inoculation on leaf gas exchange and chlorophyll contents of sugarcane genotypes.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eTreatment\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePhotosynthetic Rate\u003c/p\u003e\n\u003cp\u003e(\u0026micro;mol m\u003csup\u003e\u0026minus;\u0026thinsp;2\u003c/sup\u003e\u0026nbsp;s\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eTranspiration Rate\u003c/p\u003e\n\u003cp\u003e(mol m\u003csup\u003e\u0026ndash;2\u003c/sup\u003e\u0026nbsp;s\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eStomatal Conductance\u003c/p\u003e\n\u003cp\u003e(mol m\u003csup\u003e\u0026ndash;2\u003c/sup\u003e\u0026nbsp;s\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eIntrinsic Water-Use Efficiency\u003c/p\u003e\n\u003cp\u003e(\u0026micro;mol H\u003csub\u003e2\u003c/sub\u003eO m\u003csup\u003e\u0026minus;\u0026thinsp;2\u003c/sup\u003e\u0026nbsp;s\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eTotal Chlorophyll\u003c/p\u003e\n\u003cp\u003e(mg g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u0026nbsp;FW)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eChlorophyll \u003cem\u003ea\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e(mg g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u0026nbsp;FW)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eChlorophyll \u003cem\u003eb\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e(mg g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u0026nbsp;FW)\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eInoculated Resistant\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCPF-237\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.87 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.78 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.032 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e199.56 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.27 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.89 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.39 b\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eHSF-240\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.05 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.50 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.027 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e174.40 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.49 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.96 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.43 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCPF-243\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.69 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.66 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.038 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e139.24 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.99 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.61 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.29 d\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eInoculated Susceptible\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCP 43\u0026thinsp;\u0026minus;\u0026thinsp;33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.27 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.54 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.027 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e209.87 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.98 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.71 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.31 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSPF-245\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.47 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.45 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.032 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e135.43 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.09 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.69 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.32 c\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eHSF-240\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.77 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.38 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.026 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e177.01 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.67 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.38 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.25 e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eUn-Inoculated Resistant\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCPF-237\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7.21 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.00 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.038 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e179.24 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.57 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.09 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.43 b\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eHSF-240\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.01 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.72 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.037 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e149.42 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.87 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.34 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.57 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCPF-243\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.36 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.88 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.050 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e120.25 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.98 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.71 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.34 d\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eUn-Inoculated Susceptible\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCP 43\u0026thinsp;\u0026minus;\u0026thinsp;33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.79 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.65 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.032 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e169.40 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.19 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.81 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.39 c\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSPF-245\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.09 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.57 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.036 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e127.45 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.23 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.89 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.41 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eHSF-240\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.27 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.54 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.037 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e147.24 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.87 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.61 f\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.33 d\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"8\"\u003eValues in the column are mean of each variety and reaction type while lowercase letters were significantly different at \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.05 by Tukey\u0026rsquo;s HSD test.\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eWhip smut is one of the most prevalent and destructive diseases affecting sugarcane in almost all sugarcane growing regions of the worldwide. It possesses a very high destructive potential, causing substantial yield losses in the susceptible sugarcane cultivars. Effective disease management typically begins with the cultural practices, such as the use of resistant varieties (Iqbal et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, in the absence of resistant varieties and the non-availability of effective non-chemical measures, reliance on chemical fungicides becomes indispensable to combat destructive plant diseases and to prevent heavy economic losses (Tahir et al. \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Usman et al. \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Therefore, the search for effective fungicides is an ongoing process, as resistance can develop in targeted pathogens and new pathotypes may emerge. In regions considered hot spots for whip smut disease, the application of fungicides for seed setts treatment is a common practice. This study evaluated a wide range of fungicides against whip smut on susceptible variety of sugarcane\u003c/p\u003e\u003cp\u003eCultivation of resistant sugarcane varieties is the most effective and environmentally friendly strategy for managing sugarcane smut disease but if disease appeared in the field in epidemic form then farmers have no option except to use synthetic fungicides. So in contemporary study a number of fungicide were evaluated against whip smut of sugarcane to document their effectiveness. Ten fungicides namely Amistar Top, Triton, Epoxiconazole Epic 25% SC, Tilt, Consist 500 SC, Vibrance Duo, Aliette, Capnazale, CabrioTop, and Revus Start Pipte 430 WG were tested under laboratory conditions. Results indicated that Aliette, Tilt, and Amistar Top were particularly effective in disease management. Aliette, containing Fosetyl-Al (Alkyl Phosphonate) 800g/kg as the active ingredient, was the most efficient. It is known to inhibit fungal growth and spore germination, particularly when combined with other active ingredients. Fosetyl-Al is rapidly absorbed through both leaves and roots and exhibits both acropetal and basipetal translocation, enhancing the plant's natural defense mechanisms against fungal pathogens (Silva et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Results of the existing study are in line with Rajput et al. (\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), who demonstrated that fungicides such as Bayleton, Bavistan, and Tilt effectively controlled whip smut when applied as hot water dips. These fungicides were highly effective in reducing disease incidence and enhancing sugarcane growth parameters. Treating seed pieces (setts) with fungicides before planting significantly enhances germination and results in healthier crop stands. Fungicides work by binding to β-tubulin polymers in pathogens, inhibiting essential cellular processes like mitosis, meiosis, and cell shape maintenance (Nene and Thapliyal \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e1982\u003c/span\u003e; Horst \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). This disruption of fungal development leads to effective disease management.\u003c/p\u003e\u003cp\u003eTilt, containing Propiconazole as its active ingredient, acts as an inducer of a group of CYPs and an inhibitor of CYP51. According to Nesnow (\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), Propiconazole activates nuclear receptors that lead to the induction of various CYPs. Amistar Top, which contains azoxystrobin and difenoconazole, provides broad-spectrum activity against a range of diseases. Azoxystrobin offers anti-sporulant activity, while difenoconazole is absorbed by the plant and acts during fungal pathogen penetration and haustoria formation. Our results corroborate these mechanisms, as Tilt and Amistar Top significantly reduced disease incidence and improved plant health in the present experiments.\u003c/p\u003e\u003cp\u003eOur findings align with Abera et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), who revealed that Tilt, Bayfidan, Bayleton, and Vincit were the most effective treatments for whip smut. Sundravadana et al. (\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) also effectively controlled the disease using Propiconazole (Tilt) and Triademifon (Bayleton). Bharathi (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) recommended treating seed setts with triademifon at 0.1% for 4 hours or propiconazole at 0.1% before planting to eradicate sugarcane smut. Bhuiyan et al. (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) Understanding of environmental factors is essential for developing effective strategies to mitigate sugarcane smut disease. As climate change is projected to alter temperature and precipitation patterns, there is a growing urgency to adapt agricultural practices to minimize disease risks. By integrating knowledge of these environmental dynamics into disease management strategies, we can enhance the resilience of sugarcane crops against smut and ensure sustainable agricultural production (Yuan et al. \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Atiq et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eEnvironmental conditions play a critical role in the development and spread of the pathogen causing smut disease in sugarcane. According to Que et al. (\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2014\u003c/span\u003e), airborne teliospores are the primary source of infection in sugarcane-growing regions. Variations in disease severity can be largely attributed to environmental factors, including maximum temperature, minimum temperature, relative humidity, wind speed, and rainfall. Our study investigated these factors in relation to varietal reactions across different sugarcane varieties, revealing statistically significant correlations.\u003c/p\u003e\u003cp\u003eBoth maximum and minimum temperatures showed a significant negative correlation at P\u0026thinsp;\u0026le;\u0026thinsp;0.05% level with disease incidence, indicating reduced disease severity incidence as temperatures increase. This observation is consistent with study of Hidayah et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) highlighted the critical influence of temperature on teliospore germination and fungal growth. Hidaya et al. reported a notable reduction in disease incidence to 5.7% at 35\u0026deg;C, with peak incidence recorded between 19.3\u0026deg;C and 31.0\u0026deg;C, identifying these temperature ranges as optimal for disease development. Mansoor et al. (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) corroborated these findings, emphasizing the role of temperature in influencing smut disease dynamics.\u003c/p\u003e\u003cp\u003eSimilarly, wind speed demonstrated a significant positive correlation (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) with disease incidence, indicating that higher wind speeds enhance the dispersal of smut teliospores and increased disease likelihood in fields. Mansoor et al. (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) found maximum disease incidence (92%) occurring at wind speeds ranging 3.0 km/h, further supporting the impact of wind dynamics on smut epidemiology.\u003c/p\u003e\u003cp\u003eRelative humidity and rainfall also exhibited significant positive correlations with whip smut development. Higher relative humidity provides optimal conditions for teliospore germination, with disease incidence increasing as humidity levels rise. Sreeramulu et al. (1971) noted maximum spore dispersal at relative humidity levels of 60 to 70%, consistent with our findings. Mansoor et al. (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) also found that relative humidity influences disease incidence, underscoring the importance of moist conditions for disease progression.\u003c/p\u003e\u003cp\u003eWhen a pathogen attacks, plants prioritize the biosynthesis of defense-related substances while reducing other (e.g., growth-related) cellular activities (Usman et al. \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). This permits a reduction in physiological parameters until pathogenic growth has been halted (Bolwell and Wojtaszek \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e1997\u003c/span\u003e). This could help the plant by depriving biotrophic pathogens of nutrition. Various physiological parameters, such as chlorophyll contents (a, b, and total), photosynthetic rate, transpiration rate, stomatal conductance, and intrinsic water-use efficiency, were studied in six different genotypes (three resistant and three susceptible) of sugarcane after artificial inoculation. The tested genotypes showed a reduction in their activities due to Smut inoculation. However, the level of reduction was genotype-specific. Susceptible genotypes were found to be worse affected due to Smut inoculation, as the highest reduction in total chlorophyll, chl a, and chl b, stomatal conductance, transpiration rate, and photosynthetic activity were registered in these genotypes. The results of the current study agree with the findings of (Guo et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2005\u003c/span\u003e), who found a significant reduction in chlorophyll content in infected plants with the turnip mosaic virus. Probably, declining chlorophyll content per unit area of leaf, stomatal closure, and reduced stomatal closure upon infection are the reasons for a reduced photosynthetic rate (Shad et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Sayed \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Berova et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). The photosynthetic rate in diseased plants is reduced due to destruction of green leaf tissues and defoliation of leaves that leads to decrease in photosynthetic area and ultimately reduce photosynthesis (Cheaib and Nabil \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Similarly, various photosynthetic parameters like maximum photosynthetic rate, light saturation point, carboxylation efficiency, light compensation point, and dark respiration point in resistant sugarcane varieties was high in resistant plants as compared to susceptible varieties inoculated with pathogens (Hu et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe rate of transpiration in infected sugarcane plants was less due to reduction in maximal stomatal conductance compared to healthy. Fungal pathogens affect stomatal behavior by changing hormonal signaling pathways including abscisic acid (ABA) which controls stomatal opening and closing (Hajji et al. 2009). Susceptible plants at early stage of infection cause opening of stomata due to attack of pathogens that lead to higher transpiration rates while resistant plants contained pathogens through hypersensitive response (HR) and callose deposition which prevent further spread of bacterial pathogens and close their stomata as a defense mechanism which reduce rate of transpiration (Shad et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Jeyaraj et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Bacterial pathogens in infected plants manipulate stomata to remain open, resulting in increased stomatal conductance during early-stage infection (Ahmad et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Smut fungi secrete effector proteins that suppress salicylic acid signaling and prevent closure of stomata (Atiq et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). During later stage of infection stomatal conductance decreased due to necrosis and damaging of leaf tissues which reduce number of functional stomata (Zeng et al. \u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Resistant plants exhibit controlled stomatal conductance in response to bacterial infection and when threat is neutralized, plant reopen its stomata to restore normal photosynthesis and gas exchanges (Underwood et al. \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Water use efficiency (WUE) in inoculated plants was low due to production of toxins that damage plant cells and disrupt metabolic processes. Smut fungi also secrete enzymes like cellulases and pectinases, which degrade plant cell walls, further reducing photosynthetic efficiency and WUE.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eAmong ten tested fungicides Aliette, Tilt and Amistar top expressed promising results towards whip smut of sugarcane while in case of environmental factors maximum and minimum temperature expressed significant negative correlation while relative humidity, wind speed and rainfall displayed significant positive correlation with incidence of whip smut of sugarcane. Maximum whip smut incidence was observed at 31\u003csup\u003eo\u003c/sup\u003eC (maximum) and 19.3\u003csup\u003eo\u003c/sup\u003eC as minimum temperature, rainfall 6.6 mm, wind speed 3 km/h and 37.8 to 68% relative humidity. Smut inoculation tended to reduce the photosynthetic rate, transpiration rate, stomatal consuctance, and leaf chlorophyll (total, \u0026ldquo;a\u0026rdquo;, and \u0026ldquo;b\u0026rdquo;) content in all resistance and susceptible genotypes statistically; however, water use efficiency expressed the reverse trend in response to fungal inoculation.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eCompeting Interests\u003c/h2\u003e\u003cp\u003eAuthors have declared that no competing interests exist.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eWriting\u0026mdash;original draft, Methodology, Investigation, Formal analysis, Data collection, Conceptualization, RNN, AN, MU and MW Writing \u0026ndash; review \u0026amp; editing, Supervision, NAR, MA and AR. Writing \u0026ndash; review \u0026amp; editing, Visualization, validation, Formal analysis and references, MUQ, HA and SI. All authors reviewed the manuscript and agreed for publication.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e\u003cp\u003eWe sincerely appreciate the financial support given by the Sugarcane Research \u0026amp; Development Board Punjab Pakistan under the (Project No. 23\u0026thinsp;\u0026minus;\u0026thinsp;16). Our gratitude extends to Dr. Joseph Juma Mafurah from Egerton University for his critical review of the manuscript.\u003c/p\u003e\u003ch2\u003eData Availabilitys\u003c/h2\u003e\u003cp\u003eAll the data is available in MS.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbera T, Firehun Y, Solomon B (2009) Review of sugarcane protection research in Ethiopia. Inc Crop Prod Imp Plant Prot 2: 409\u0026ndash;447.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAbera T (2001) Review of reaction of sugarcane varieties to smut (\u003cem\u003eUstilago scitaminea\u003c/em\u003e Syd.) in Ethiopia. Review of sugarcane research in Ethiopia: II. Crop Protection (1970\u0026ndash;1998, pp. 1\u0026ndash;30) Ethiopian Sugar Industry Support Center Research and Training Service, Wonji, Ethopia.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAgnihotri VP (1990) Smut of Sugarcane. Diseases of sugarcane and sugarbeet (Revised Ed., pp. 73\u0026ndash;76). Oxford and IBH Publishing Co. 66 Janapath, New Dehli, India.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAhmad H, Rajput NA, Atiq M, Kachelo GA, Usman M, Tariq H, Wahab M (2024) Detection of \u003cem\u003ePhytophthora nicotiana\u003c/em\u003e induced citrus gummosis by the loop mediated isothermal amplification. Pak J Bot 56(5): 1\u0026ndash;12.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAlbert H, Schenck S (1996) Development of a sensitive assay for detection of sugarcane smut disease caused by \u003cem\u003eUstilago scitaminea\u003c/em\u003e. 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Current Opinions Biotechnol 21(5): 599\u0026ndash;603.\u003c/span\u003e\u003c/li\u003e\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":"Saccharum officinarum, Leaf Gasses Exchange, Smut, Management, Epidemiology","lastPublishedDoi":"10.21203/rs.3.rs-7279829/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7279829/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSugarcane, a cornerstone of Pakistan's agricultural economy, standing as the second-largest cash crop in the country. The pervasive threat of whip smut, caused by \u003cem\u003eSporisorium scitamineum\u003c/em\u003e, poses a formidable challenge, often resulting in yield losses ranging from 18\u0026ndash;68%. This study explores the incidence of whip smut in sugarcane across Faisalabad, investigating its correlation with environmental factors such as maximum and minimum temperatures (\u0026deg;C), relative humidity (%), rainfall (mm), and wind speed (Km/h), disease impact on plant physiology and effective management strategies using synthetic chemicals. Field experiments, incorporating artificial inoculation across ten sugarcane varieties, revealed varying susceptibility, with HSF-242 exhibiting the highest disease incidence (92%) and CP43-33 the lowest (78%). Statistical analyses highlighted significant negative correlations between disease incidence and maximum/minimum temperatures, while positive correlations were observed with relative humidity, rainfall, and wind speed. \u003cem\u003eIn vitro\u003c/em\u003e evaluations of ten chemical fungicides identified Aliette (30%), Tilt (40%), and Amistar Top (44.67%) as the most effective in suppressing \u003cem\u003eS. scitamineum\u003c/em\u003e. Subsequent field trials confirmed Aliette's superior efficacy, demonstrating its potential in disease management strategies. Photosynthetic rate, transpiration rate, stomatal conductance, and chlorophyll contents (total, a, and b) were observed high in healthy plants as compared to inoculated ones. While water use efficiency was observed in greater amounts in inoculated plants as compared to healthy plants. This study contributes novel insights into managing sugarcane whip smut, offering practical strategies to mitigate economic losses and sustainably enhance crop yields in global sugarcane production systems.\u003c/p\u003e","manuscriptTitle":"Physiology, Epidemiology and Fungicidal Subdual Strategies for whip smut of sugarcane Caused by Sporisoriums Scitamineum","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-03 14:40:44","doi":"10.21203/rs.3.rs-7279829/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":"ead95b09-a7e8-4c76-a6e0-6d4f1d649e67","owner":[],"postedDate":"September 3rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-30T13:38:34+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-03 14:40:44","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7279829","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7279829","identity":"rs-7279829","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}