Riboflavin (Vitamin B2)-induced biomass, yield and antioxidant potential of Bittergourd in water-deficit condition

Riboflavin (Vitamin B2)-induced biomass, yield and antioxidant potential of Bittergourd in water-deficit condition | 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 Riboflavin (Vitamin B2)-induced biomass, yield and antioxidant potential of Bittergourd in water-deficit condition Ammara Razzaq, Naila Ali, Muhammad Iftikhar, Anis Ali Shah, Zaib-un-Nisa ., and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5356796/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 Drought is an abiotic stress for many species that live in soil. Although many plants have been studied for drought-adaptive processes, but a little is known about Bittergourd ( Momordica charantia L. ). Antioxidants are especially important because desiccation causes an increase in the intracellular generation of reactive oxygen species. Riboflavin (vitamin B2) is one antioxidant that influences plant drought tolerance. It causes plants to produce reactive oxygen species(ROS) when exposed to light, is an excellent photosensitizer for biocidal reactions. This study explores the possible protective role of riboflavin (0, 25, 50, 100 ppm) foliar application against drought stress (i.e., 60% and 100% field capacity) in bitter gourd plants. A pot experiment was conducted in a completely randomized design (CRD) with four replicates during spring season of 2021 at the field area of University of Lahore. Results indicated that the foliar applications of riboflavin (RF) improved morphological and physiological attributes in plants as compared to control plants when grown under drought. Although with increasing concentration of riboflavin all the parameters gradually increased. Fruit count (44%), weight (20%), diameter (22%) and volume (43%) also increased with foliar application. Photosynthetic pigments (Chlorophyll a by 29%, Chlorophyll b by 48%, Total chlorophyll by 34% and Carotenoid by 16%) and metabolites (Total free amino acids by 41% and Total soluble proteins by 37%) increased under the influence of riboflavin. Moreover, Phenolic contents and antioxidant activities (Catalase, Peroxidase and DPPH) of bitter gourd were highlighted showing significant increase in stress which further enhanced under the effect of riboflavin by 23%, 42%, 36% and 56%, respectively to determine the specific direction to be taken for future plant antioxidant research. Figures Figure 1 Figure 2 Figure 3 Introduction Drought being a stressor for soil-dwelling organisms including plants leading to reduction in crop yields globally​[1,2​]. Plants exhibit varying degrees of vulnerability to drought stress at various stages of their life cycles. Early in seedling development, most crops exhibit less sensitivity while, water scarcity causes numerous physiological changes in plants during their generative stage, which lowers plant output​[3]. Many physiological features of plants alter by drought, like leaf morphology, enzyme activity, water and mineral intake, photosynthetic and transpiration rates, and stomatal conductance leading to reduced agricultural production and yield​ [4]​. The function of important metabolic processes like photosynthesis, mineral absorption, and assimilation also changed as a result of drought stress due to the excessive formation of reactive oxygen species (ROS) that causes oxidative damage to plants​[1]​. In order to neutralize the excess of ROS and protect metabolism, plants up-regulate their antioxidant defenses and accumulate osmolytes​[5​]. The growth and development of plants are sped up by a variety of growth-promoting chemicals, which also enable plants to survive in challenging environments​[6–8]​. Recent research has revealed that vitamins have a dual purpose in addition to being essential nutrients. Vitamins, particularly ascorbic acid and thiamine, are important for reducing environmental stress. In order to increase crop productivity and resilience to biotic and abiotic challenges, plant-derived vitamins control plant metabolism, alter redox chemistry, and serve as enzymatic cofactors​[9,10​]. The group of vitamin-B containing cobalamin, niacin, and riboflavin have been said to reduce environmental stress, although their precise mechanism of action is still unknown​ [11]​. The precursor to many necessary cofactors for different metabolic pathways is riboflavin. Plants, bacteria and fungi, can de novo synthesize riboflavin unlike animals, through a mechanism that has been preserved from ancestors. In the course of osmotic stress, riboflavin is known to function as a cofactor in both antioxidation and peroxidation​[12]​. However, the mechanism of riboflavin to control seed formation is still not known​[13]​. Even very little amounts of riboflavin increased the tobacco plant's ability to withstand drought when grown under natural conditions​[14]​. It is a potent antioxidant that accumulates antioxidant chemicals in a variety of plant cells showing its ability as an electron acceptor​[15]​. It is crucial for the cell's oxidative process​[16]​. By triggering the stress response in plants, riboflavin can lessen abiotic stresses​[17​]. Metabolites such acetyl CoA, 2 oxoglutarate, and the majority of B vitamins play crucial roles in numerous biochemical pathways in addition to acting as co-substrates for numerous epigenetic masters, integrating nutrition, metabolism and gene expression​[18]​. Riboflavin (Vitamin B2) generates cofactors, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), which are implicated in many metabolic processes, including electron transport chain, fatty-acid β‐oxidation, tricarboxylic acid (TCA) cycle, redox homeostasis, as well as signaling through oxidation-reduction reactions​[19]​. Guanosine triphosphate (GTP) and ribulose 5 phosphate serve as the starting points for riboflavin's production pathway​[20]​. The order of reaction steps and the fusion of riboflavin biosynthesis proteins (RIB) have undergone slight evolutionary alterations in distinct lineages​[21]​. Animals and plants also suffer severe repercussions when riboflavin is deficient, in plants weakened stress resistance and flawed seed development have been observed​[22–24​]. Although the production and metabolism of riboflavin have been extensively studied, its physiological and biological applications are particularly limited in vegetables. In our best knowledge there have been limited study found about role of riboflavin in vegetables. In this piece of work, the potential benefits of riboflavin foliar treatment in alleviating drought stress in Momordica charantia L. have been investigated. Materials and Methodology On bitter gourd, experiment was carried out at the IMBB greenhouse at The University of Lahore, Lahore, to ascertain the functions of riboflavin (0, 25, 50, and 100 ppm) in reducing drought stress (100% and 60% FC). From the Ayub Agriculture Research Institute (AARI) in Faisalabad, bitter gourd seeds were gathered. Five seeds of bittergourd were planted in pots with 7 kg of thoroughly cleansed loamy soil. Plant growth and fruit attributes Manual measurements of the length of the shoot and the root were made using a scale. Fresh weight of the shoot, the root, and the fruit were also taken along with fruit and leaves count, leaf area, fruit diameter and volume of each replication. To record the data for the dry weights of the shoot and root, samples were oven-dried separately at 60°C until the stable mass was reached. Determination of photosynthetic contents Following Arnon's method (1949)​[25]​, the contents of chlorophyll ( a, b , and total) and carotenoids were determined. Freshly harvested shoots(5g) were crushed in 10 ml of acetone (80%). A spectrophotometer was used to measure the filtrate's absorbance at 480, 645, and 663 nm, separately, after the filtrate was collected. Determination of metabolites The total phenolic content was ascertained using the method of Lamuela​[26]​ with Folin–Ciocalteau phenol reagent. The absorbance was measured at 750 nm. The total phenolic content was determined as milligrams per gram of fresh leaf. The concentration of protein in the leaf extracts was measured following the method of Bradford (1976)​[27]​. About 2 ml of Bradford reagent and 0.1 ml of the leaf extracts were taken into test tubes and the mixture kept at room temperature for 5 min. The absorbance of each sample was measured at 595 nm. The method of Hamilton and Van Slyke (1943)​[28​] was followed to measure the total free amino acids. Results were found by mixing 10% pyridine and 1% ninhydrin with 1 ml of leaf extract, keeping the test tubes at room temperature for 30 minutes, and construing the optical density at 570 nm. Determination of antioxidant enzyme activities Fresh shoots (0.5 g) were crushed in buffer (50 mM, pH 7.8) on an ice bath. The mixture was then centrifuged for 20 minutes at 4°C using 15,000 rpm. The activity of antioxidant enzymes was assessed using the obtained supernatants. To measure the catalase (CAT) and peroxidase (POD) activities in bitterguord, the method of Chance and Maehly (1955)​[29​] was used. After 20 seconds, fluctuations in absorbance at 470 nm were observed for 3 minutes. A change in absorbance of 0.01 U/min was considered to be one unit of enzymatic activity. The DPPH activity was measured using the technique described by Valko​[30]​. After an incubation of the samples for 30min in dark, the absorbance was measured at 517nm wavelength. Enzyme activity was determined on the basis of soluble protein. Statistical analysis With the aid of computer-based software (Statistix version 8.1), analysis of variance (ANOVA) was performed for each parameter. The data was represented graphically using Microsoft Excel. Results Riboflavin foliar application enhanced growth indices in bitter gourd Statistical analysis showed significant differences in plant height as well as root and shoot fresh and dry biomass (Table. 1). Drought reduced the root length by 27%, shoot length by 22%, root fresh and dry biomass by 31.2% and 53% while shoot fresh and dry biomass displayed a significant drop by 19.25% and 17% respectively, as compared to fully irrigated plants. Drought conditions also dropped number of leaves by 15% and leaf area by 20%. Riboflavin application under drought displayed positive effect on bitter gourd growth and biomass at all applied concentrations as plants exhibited a significant incline in root length by 18% at 25 ppm, 33% at 50 ppm and 40% at 100 ppm riboflavin concentration as compared to only drought treated plants. Similarly, bitter gourd shoot displayed improved growth by 12.8%, 25.4% and 35% at 25 ppm, 50 ppm and 100 ppm concentrations of riboflavin respectively, as compared to drought treated plants. Plant fresh and dry biomass also followed the similar trend and highest increase in root fresh and dry biomass was calculated at 100 ppm riboflavin by 63–66% while shoot fresh and dry biomass was improved by 40–42% as compared to only drought treated plants. Further, bitter gourd plants exhibited a significant incline in number of leaves by 14% at 25 ppm, 29% at 50 ppm and 37% at 100 ppm as well as leaf area increased by 17% at 100 ppm riboflavin concentration as compared to only drought treated plants. As shown in Figure S1 , the effect of riboflavin was significant on plant growth. Riboflavin foliar spray improved fruit attributes in bitter gourd Significant differences were observed in yield attributes such as fruit count, fruit weight, fruit volume and fruit diameter (Table. 2). Drought reduced the fruit weight by 20%, number of fruits by 44%, fruit diameter by 22%, fruit volume by 43% as compared to fully irrigated plants. Riboflavin application under both control and drought conditions displayed positive effect on bitter gourd growth at all applied concentrations. Similarly, bitter gourd fruit weight displayed improved growth by 20%, 39.6% and 51.48% at 25 ppm, 50 ppm and 100 ppm concentrations of riboflavin respectively, as compared to drought treated plants. Number of fruit also followed the similar trend and highest increase in number of fruit was calculated at 100 ppm riboflavin by 28–34% while fruit diameter and fruit volume was improved by 40% and 42% calculated at 100 ppm concentration of riboflavin respectively as compared to only drought treated plants. Riboflavin foliar spray significantly changed fruit count, size and weight properties. Riboflavin foliar spray enhanced photosynthetic contents in bitter gourd Chlorophyll a , chlorophyll b , total chlorophyll and carotenoids showed positive responses with the increase of riboflavin as determined by statistical analysis (Fig. 1 ). Drought reduced the chlorophyll a by 24%, chlorophyll b by 35%, total chlorophyll by 28%, and carotenoid by 24.35%, as compared to fully irrigated plants. Riboflavin foliar spray under drought displayed positive effect on bitter gourd growth at all applied concentrations as plants exhibited a significant incline in chlorophyll a by 25% at 25 ppm, 26% at 50 ppm and 29% at 100 ppm riboflavin concentration as compared to only drought treated plants. Similarly, bitter gourd displayed increase contents of chlorophyll b by 22.44%, 43% and 48% at 25 ppm, 50 ppm and 100 ppm concentrations of riboflavin respectively, as compared to drought treated plants. Total chlorophyll also followed the similar trend and highest increase was calculated at 100 ppm riboflavin by 30–34% while carotenoids 16% was calculated at 100 ppm concentration of riboflavin respectively as compared to only drought treated plants. Riboflavin application under control conditions also exhibited positive effects on all above mentioned parameters. These results suggest, positive impact of riboflavin on plant pigments. Riboflavin foliar spray concerted regulation of metabolites in bitter gourd Plant metabolites such as total free amino acids, total soluble proteins and total phenolics showed positive responses with the increase of riboflavin concentration both under control and drought conditions (Fig. 2 ). Drought reduced the total free amino acids by 36.70% and total soluble protein by 30.25% respectively, as compared to fully irrigated plants. Riboflavin application under drought displayed positive effect on bitter gourd metabolites at all applied concentrations as plants exhibited a significant incline in total free amino acids by 30% at 25 ppm, 33% at 50 ppm and 41% at 100 ppm riboflavin concentration as compared to only drought treated plants. Similarly, bitter gourd total soluble proteins displayed improved contents by 15%, 28% and 37% at 25 ppm, 50 ppm and 100 ppm concentrations of riboflavin respectively, as compared to drought treated plants. Furthermore, drought increased the total phenol content by 24%, as compared to fully irrigated plants while under riboflavin application, plants further exhibited a significant incline in total phenols by 19% at 25 ppm, 20% at 50 ppm and 23% at 100 ppm riboflavin concentration as compared to only drought treated plants. Riboflavin foliar spray enhanced antioxidant enzyme activities in bitter gourd Catalase, peroxidase and DPPH assay showed positive response with the increase of riboflavin concentrations under both control and drought conditions as mentioned by statistical analysis (Fig. 3 ). Drought conditions increased the catalase activity by 24.28%, peroxidase activity by 27.68% and DPPH assay by 51% as compared to fully irrigated plants. Riboflavin application under drought exhibited a significant decline in catalase by 11% at 25 ppm, 22.11% at 50 ppm and 42% at 100 ppm riboflavin concentration as compared to only drought treated plants. Similarly, bitter gourd peroxidase displayed decrease in enzyme activities by 17%, 27.47% and 36.50% at 25 ppm, 50 ppm and 100 ppm concentrations of riboflavin respectively, as compared to drought treated plants. DPPH assay also followed the similar trend and highest increase was calculated at 100 ppm riboflavin by 49–56% concentration of riboflavin respectively as compared to only drought treated plants. These results strongly suggest that riboflavin foliar application strengthen the defense system of bitter gourd. Discussion Plants react to various stressors through a variety of biological, morphological, and molecular pathways, when dealt with stress​[31]​. Many organisms including plants that live in soil are stressed by drought. Since desiccation is accompanied by an excessive intracellular generation of reactive oxygen species, antioxidants are particularly important. One antioxidant that controls a plant's ability to withstand drought is riboflavin (vitamin B2)​[32​]. The main function of water-soluble vitamin B2 (riboflavin) in cell biology is related to its conversion into FMN and FAD, the cofactors of numerous dehydrogenases, oxidases, and reductases engaged in a wide range of biological activities, including energetic metabolism and chromatin remodeling​[33]​. The production of vegetables is significantly impacted by drought. Drought stress has an impact on a number of plant functions, including photosynthesis, transpiration, the transfer of resources from source to sink, etc. Stomata are closed and leaf area is decreased in times of water scarcity, which lowers photosynthetic and transpiration activity and reduces water use​[34​]. Additionally, it slows down the physiological, biochemical, and morphological processes that contribute to plant growth. The primary responses of plants to stress include stunted growth of roots and shoots, sluggish rate of germination, closure of stomata, reduction in seedling growth, and worsening of photosynthetic activity​[35​]. In the current study, water stress had a noticeable impact on the morphological characteristics of plants. When compared to plants produced in a drought-prone environment, these metrics are much higher in plants grown under normal conditions. The findings of the current work are consistent with the results of Talebi and colleagues​[36​], who reported similar findings about Gazania plants. In accordance with their findings, the application of riboflavin at various concentrations improved the vegetative development metrics. The foliar application of riboflavin is crucial for the metabolic, biochemical, and physiological processes of plants (such as nutrient absorption, water relations, photosynthesis, and stress signaling, etc.), which ultimately have a positive impact on plant growth, yield, and crop quality as well as vegetative and reproductive development​[37​]. ABA, also known as the stress hormone, is released in the roots and then goes to the plant's above-ground sections, where it sends signals for defenses against drought stress. Recently, a number of molecular mechanisms have been unveiled to determine the effect of hormones on ABA signal transduction genes and transcriptional profiles of stress related genes​[38,39]​. In pursuit of water, roots begin penetrating soils farther. Fruit failure results from decreased plant activity, which causes flowers to set fruit poorly or not at all[34]. Similar findings were made when working on bittergourd affected by drought; the fruit count along with its morphological parameters decreased, while, the application of riboflavin enhanced the aforementioned characteristics. The future depends in finding ways to combat the water shortage and drought stress that would guarantee abundant vegetable crop production to feed a growing population​[34]​. Under conditions of water stress, plants are less able to absorb nitrogen from the soil, which results in a decrease in plant biomass and nitrogen-related chemical components like total free amino acids and sugar​[40]​. In the current study, riboflavin considerably outperformed plants that were not supplemented with it in terms of total soluble proteins and total free amino acids under water stress circumstances. These outcomes are consistent with those reported by Thompson and colleagues​[41]​, who applied riboflavin topically to Nicotiana tabacum and saw comparable outcomes. Riboflavin improved the manufacture of amino acids such proline, glycine, serine, leucine, and lycine in Vitis vinifera , according to a subsequent study by Cangi and his colleagues​[42​]. As a molecular chaperone, proline works. The effects of the drought on crop productivity and the viability of agriculture were undesirable. It has a significant impact on the morphological and biochemical processes in plants, which inhibit growth and change enzyme activity at the cellular level​[43]​. Uncontrolled free radicals, increased, and plants use enzymatic and non-enzymatic antioxidants to reduce oxidants and manage cellular homeostasis under oxidative stress situations​[44​]. Severe oxidative stress can result from ROS buildup in plants​[45​]. The enzymatic antioxidant enzymes work together to counteract the excessive ROS production in plants, defending the structures and functions of cellular constituents. Generally speaking, under various abiotic stress conditions, the activity of several antioxidant enzymes in plants rises​[46–48]​. According to Heidari​[49]​, an increase in enzymatic antioxidant activity is correlated with an increase in stress tolerance. When compared to normal plants, the antioxidant activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), catalase (CAT), and peroxidase (POD) were found to be improved by drought in the current study. These results are in line with those of Akram and his colleagues​[50​] (2012), who reported that drought stress caused a significant buildup of H 2 O 2 in leaves and MDA increased the activity of SOD, CAT, and POD in sunflower plants. According to several studies, the antioxidants catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), glutathione peroxidase (GPX), and polyphenol oxidase (APX) are among those that can be activated by riboflavin treatment in Hibiscus sabdariffa L.​[51] ​ . Conclusion Plant research now prioritizes studying how to preserve food production and how plants can withstand drought. All types of crops, including field crops, vegetable crops, fruit crops, spices, and others, are negatively impacted by drought. By decreasing both leaf area and photosynthetic rate per unit leaf area, drought stress is known to limit photosynthesis. It is essential to study the antioxidant enzyme research to comprehend the molecular mechanisms of various stress responses in bittergourd in order to increase crop production in controlled and water-deficit environments with the administration of vitamin B2 (riboflavin). Consequently, to lessen the negative effects of drought- generated oxidative stress, the findings showed a distinct pattern of antioxidants, where stimulation of DPPH, POD, and CAT activity was observed to be more effective in riboflavin-treated plants than in stressed plants. Because ROS created under stressful conditions can seriously damage the mechanical assembly of the photosynthetic system. Therefore, the use of riboflavin improved the morphological, biochemical and antioxidant activities of bitterguord under the influence of water scarcity. Abbreviations Reactive Oxygen Species (ROS);; Ascorbic Acid, (ASC); Superoxide Dismutase, (SOD); Catalase, (CAT), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and peroxidase (POD), superoxide dismutase (SOD), glutathione peroxidase (GPX). polyphenol oxidase (APX). Declarations Funding: Researchers supporting the project (RSP2024R393) at King Saud University, Riyadh, Saudi Arabia. . Conflict of interest: All authors declare no conflict of interest Authors Contributions: [AR] conceived the study and design the experiment. [NA] added valuable suggestions during experiment and manuscript improvement. [ZUN] contributed various inputs during the manuscript preparation and supervision. [MYA], [TF] performed data analysis and design the manuscript. Validation, funding contributed by [VR]. [MAES] and [MI] involved in drafting article, revision, arranged the figures, tables, experiment validation and manuscript description. [AAS] approved the final version to be published. All the authors read and approved the finalized manuscript for publication. Ethics approval and consent to participate Not applicable Consent for publication All authors give permission to the publisher to publish this research work. Availability of data and materials All data is presented in this manuscript. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. Competing interests The authors declare no known competing financial interest. 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Zaid, A. & Mohammad, F. Methyl jasmonate and nitrogen interact to alleviate cadmium stress in menthaarvensis by regulating physio-biochemical damages and ROS detoxification. J Plant Growth Regul. 37, 1331–1348 (2018). Zaid, A. & Wani, S. H. Reactive Oxygen Species Generation, Scavenging and Signaling in Plant Defense Responses. Bioactive Molecules in Plant Defense. (Springer, ; pp. 111–132, 2019). Zaid, A., Mohammad, F. & Fariduddin, Q. Plant growth regulators improve growth, photosynthesis, mineral nutrient and antioxidant system under cadmium stress in menthol mint (Menthaarvensis L.) Physiol Mol Biol Plants. 26, 25–39 (2020). Heidari, P., Amerian, M. R. & Barcaccia, G. Hormone profiles and antioxidant activity of cultivated and wild tomato seedlings under low-temperature stress. Agronomy. 11, 1–16 (2021). Akram, N. A., Ashraf, M. & Al-Qurainy, F. Aminolevulinic acid-induced changes in some key physiological attributes and activities of antioxidant enzymes in sunflower (Helianthus annuus L.) plants under saline regimes. Sci Hortic 142, 143–148 (2012). Abdellatif, Y. M. R. & Ibrahim, M. T. S. Non-enzymatic anti-oxidants potential in enhancing Hibiscus sabdariffa L. tolerance to oxidative stress. International Journal of Botany 14, 43–58 (2018). Tables TABLE 1: The effect of foliar application of Riboflavin on growth indices of bitter gourd under drought stress. Condition Foliar ppm Root length (cm) Shoot Length (cm) Root fresh weight(g) Shoot fresh weight(g) Root dry weight(g) Shoot dry weight(g) Leaf Count (n) Leaf Area (cm 3 ) Control 0 13.6 de 220 d 2.50 e 16.1 de 0.45 ef 3.4 d 150 d 23.4 d 25 15.5 bcd 255 c 3.95 c 18.5 cd 0.84 c 4.5 bc 172 c 27.1 cd 50 17.4 b 289 b 4.72 b 22.9 b 1.10 b 4.9 b 197 b 32.2 b 100 19.4 a 325 a 5.9 a 26.5 a 1.31 a 5.8 a 217 a 36.4 a Drought 0 9.8 f 170 f 1.72 f 13.0 e 0.21 g 2.8 e 122 e 17.1 f 25 12.0 e 195 e 2.44 e 15.9 de 0.35 fg 3.4 d 147 d 21.3 e 50 14.7 cd 228 d 3.40 d 19.5 c 0.54 de 4.0 c 170 c 27.1 cd 100 16.5 b 265 c 4.73 b 22.7 b 0.63 d 4.7 b 189 b 30.1 bc ANOVA (F Value) Drought(D) *** *** *** *** *** *** *** *** Foliar(F) *** *** *** *** *** *** *** *** Drought*foliar ns ns *** ns *** ns ns ns Each value is a mean of four replicates; different alphabetic letters indicate significant differences (P≤0.05) among treatments, *, ** and *** indicated significance at P≤0.05, P≤0.01 and P≤0.001 respectively; ns indicated non-significant differences. TABLE 2: The effect of foliar application of Riboflavin on fruit attributes of bitter gourd under drought stress. Condition Foliar ppm Fruit Weight (g) Number of Fruits (n) Fruit Diameter Fruit Volume Control 0 13.6 d 6 cd 1.7 d 6.9 d 25 16.4 c 7 bc 12.1 c 9.7 c 50 21.1 b 8 b 2.5 b 12.2 b 100 26.3 a 10.7 a 3.0 a 15.3 a Drought 0 10.1 e 2.2 f 1.3 f 3.8 f 25 13.9 d 4 ef 1.6 e 5.5 e 50 17.1 c 5 de 2.0 c 7.3 d 100 20.9 b 7 bc 2.4 b 8.8 c ANOVA (F Value) Drought(D) *** *** *** *** Foliar(F) *** *** *** *** Drought*foliar ** ns ** *** Each value is a mean of four replicates; different alphabetic letters indicate significant differences (P≤0.05) among treatments, *, ** and *** indicated significance at P≤0.05, P≤0.01 and P≤0.001 respectively; ns indicated non-significant differences. Additional Declarations No competing interests reported. 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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-5356796","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":375531232,"identity":"cecd9836-1c0d-471b-9c6a-70421d38223c","order_by":0,"name":"Ammara Razzaq","email":"","orcid":"","institution":"The University of Lahore","correspondingAuthor":false,"prefix":"","firstName":"Ammara","middleName":"","lastName":"Razzaq","suffix":""},{"id":375531233,"identity":"2bf9bcc7-45c4-4f0f-89d0-34c795a53a9a","order_by":1,"name":"Naila Ali","email":"","orcid":"","institution":"The University of Lahore","correspondingAuthor":false,"prefix":"","firstName":"Naila","middleName":"","lastName":"Ali","suffix":""},{"id":375531234,"identity":"1fd14f92-e7b0-4e39-aa42-59916286d163","order_by":2,"name":"Muhammad Iftikhar","email":"","orcid":"","institution":"University of Education Lahore","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Iftikhar","suffix":""},{"id":375531235,"identity":"e0995520-83fc-4b82-b298-bcf99d4a26e5","order_by":3,"name":"Anis Ali Shah","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4UlEQVRIiWNgGAWjYFACxgYGhgKJBAYG5gNAnoQMkVoMgFrY2BJAWniItMmAAaiFxwDEJKyFf3Zz84cfBhZ5/PI9n1/dqLHgYWA/fHQDPi0Sdw62SfYYSBRLtvFus845BnQYT1raDbzW3EhsY+AxkEjccIx3m3EOG1CLBI8ZXi3yNxKbP/4Ba+F5ZpzzjwgtBjcSG6QhtvAwP85tI0KLIdBh0jJgv6SZMef2SfCwEfKL3I30xx/fVNTl8TMffvw551udHD/74WP4vY8E2CTAJLHKQYD5AymqR8EoGAWjYOQAAAfcRJ0khzWkAAAAAElFTkSuQmCC","orcid":"","institution":"University of Education Lahore","correspondingAuthor":true,"prefix":"","firstName":"Anis","middleName":"Ali","lastName":"Shah","suffix":""},{"id":375531236,"identity":"435129b9-ccdb-419e-874a-29c6e7a502e6","order_by":4,"name":"Zaib-un-Nisa .","email":"","orcid":"","institution":"The University of Lahore","correspondingAuthor":false,"prefix":"","firstName":"Zaib-un-Nisa","middleName":"","lastName":".","suffix":""},{"id":375531237,"identity":"c7986a26-b852-47bf-8f79-249f0c040e20","order_by":5,"name":"Muhammad Yasin Ashraf","email":"","orcid":"","institution":"The University of Lahore","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"Yasin","lastName":"Ashraf","suffix":""},{"id":375531238,"identity":"1b55f5b5-bcf5-42e9-bb01-035e9cb8e397","order_by":6,"name":"Tatheer Fatima","email":"","orcid":"","institution":"The University of Lahore","correspondingAuthor":false,"prefix":"","firstName":"Tatheer","middleName":"","lastName":"Fatima","suffix":""},{"id":375531239,"identity":"0382bc0e-d300-4902-8ffa-4a5ded215cf9","order_by":7,"name":"Vaseem Raja","email":"","orcid":"","institution":"Chandigarh University","correspondingAuthor":false,"prefix":"","firstName":"Vaseem","middleName":"","lastName":"Raja","suffix":""},{"id":375531240,"identity":"5c36b278-be17-4572-9a4f-6e8df193a2c1","order_by":8,"name":"Mohamed A. El-Sheikh","email":"","orcid":"","institution":"King Saud University","correspondingAuthor":false,"prefix":"","firstName":"Mohamed","middleName":"A.","lastName":"El-Sheikh","suffix":""}],"badges":[],"createdAt":"2024-10-29 20:08:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5356796/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5356796/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":68910742,"identity":"29ff429e-8659-4aab-8ef7-d63917fbe2d8","added_by":"auto","created_at":"2024-11-13 11:42:25","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":103362,"visible":true,"origin":"","legend":"\u003cp\u003eThe effect of different concentrations of foliar application of riboflavin on photosynthetic contents of bitter gourd (\u003cem\u003eMomordica charantia\u003c/em\u003e) under drought. Mean values are the average of four replicates. The upper case alphabetic letters indicate significant differences (P\u003cu\u003e\u0026lt;\u003c/u\u003e0.05) among treatments using Tukey's HSD test.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-5356796/v1/9479860c949acd210c63c9bc.jpeg"},{"id":68910744,"identity":"c9dab535-a4bb-4961-b275-0bc399cb90d0","added_by":"auto","created_at":"2024-11-13 11:42:25","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":77952,"visible":true,"origin":"","legend":"\u003cp\u003eThe effect of different concentrations of foliar application of riboflavin on metabolites of bitter gourd (\u003cem\u003eMomordica charantia\u003c/em\u003e) under drought. Mean values are the average of four replicates. The upper case alphabetic letters indicate significant differences (P\u003cu\u003e\u0026lt;\u003c/u\u003e0.05) among treatments using Tukey's HSD test.\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-5356796/v1/2b667e1de7e37c3e5918a781.jpeg"},{"id":68910870,"identity":"8041371f-9e8d-43a4-82d8-80590f327697","added_by":"auto","created_at":"2024-11-13 11:50:25","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":170806,"visible":true,"origin":"","legend":"\u003cp\u003eThe effect of different concentrations of foliar application of riboflavin on antioxidant enzymatic activites of bitter gourd (\u003cem\u003eMomordica charantia\u003c/em\u003e) under drought. Mean values are the average of four replicates. The upper case alphabetic letters indicate significant differences (P\u003cu\u003e\u0026lt;\u003c/u\u003e0.05) among treatments using Tukey's HSD test.\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-5356796/v1/d248ae752d3768861c3fad9f.jpeg"},{"id":82201478,"identity":"cd9d8fe2-53fa-43ee-b51f-eb02b3ecefcf","added_by":"auto","created_at":"2025-05-07 16:08:39","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1442278,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5356796/v1/252810fe-aa6a-4b02-bfc9-9fed5ef7fcd1.pdf"},{"id":68910746,"identity":"cc31799f-c967-46ed-85c4-2816629c4baa","added_by":"auto","created_at":"2024-11-13 11:42:25","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":2018104,"visible":true,"origin":"","legend":"","description":"","filename":"SUPPLEMENTARYFIGURE.docx","url":"https://assets-eu.researchsquare.com/files/rs-5356796/v1/49b8d3444e5431338c41d9c9.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Riboflavin (Vitamin B2)-induced biomass, yield and antioxidant potential of Bittergourd in water-deficit condition","fulltext":[{"header":"Introduction","content":"\u003cp\u003eDrought being a stressor for soil-dwelling organisms including plants leading to reduction in crop yields globally​[1,2​]. Plants exhibit varying degrees of vulnerability to drought stress at various stages of their life cycles. Early in seedling development, most crops exhibit less sensitivity while, water scarcity causes numerous physiological changes in plants during their generative stage, which lowers plant output​[3].\u003c/p\u003e \u003cp\u003eMany physiological features of plants alter by drought, like leaf morphology, enzyme activity, water and mineral intake, photosynthetic and transpiration rates, and stomatal conductance leading to reduced agricultural production and yield​ [4]​. The function of important metabolic processes like photosynthesis, mineral absorption, and assimilation also changed as a result of drought stress due to the excessive formation of reactive oxygen species (ROS) that causes oxidative damage to plants​[1]​. In order to neutralize the excess of ROS and protect metabolism, plants up-regulate their antioxidant defenses and accumulate osmolytes​[5​].\u003c/p\u003e \u003cp\u003eThe growth and development of plants are sped up by a variety of growth-promoting chemicals, which also enable plants to survive in challenging environments​[6\u0026ndash;8]​. Recent research has revealed that vitamins have a dual purpose in addition to being essential nutrients. Vitamins, particularly ascorbic acid and thiamine, are important for reducing environmental stress. In order to increase crop productivity and resilience to biotic and abiotic challenges, plant-derived vitamins control plant metabolism, alter redox chemistry, and serve as enzymatic cofactors​[9,10​]. The group of vitamin-B containing cobalamin, niacin, and riboflavin have been said to reduce environmental stress, although their precise mechanism of action is still unknown​ [11]​.\u003c/p\u003e \u003cp\u003eThe precursor to many necessary cofactors for different metabolic pathways is riboflavin. Plants, bacteria and fungi, can \u003cem\u003ede novo\u003c/em\u003e synthesize riboflavin unlike animals, through a mechanism that has been preserved from ancestors. In the course of osmotic stress, riboflavin is known to function as a cofactor in both antioxidation and peroxidation​[12]​. However, the mechanism of riboflavin to control seed formation is still not known​[13]​. Even very little amounts of riboflavin increased the tobacco plant's ability to withstand drought when grown under natural conditions​[14]​. It is a potent antioxidant that accumulates antioxidant chemicals in a variety of plant cells showing its ability as an electron acceptor​[15]​. It is crucial for the cell's oxidative process​[16]​. By triggering the stress response in plants, riboflavin can lessen abiotic stresses​[17​].\u003c/p\u003e \u003cp\u003eMetabolites such acetyl CoA, 2 oxoglutarate, and the majority of B vitamins play crucial roles in numerous biochemical pathways in addition to acting as co-substrates for numerous epigenetic masters, integrating nutrition, metabolism and gene expression​[18]​. Riboflavin (Vitamin B2) generates cofactors, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), which are implicated in many metabolic processes, including electron transport chain, fatty-acid β‐oxidation, tricarboxylic acid (TCA) cycle, redox homeostasis, as well as signaling through oxidation-reduction reactions​[19]​. Guanosine triphosphate (GTP) and ribulose 5 phosphate serve as the starting points for riboflavin's production pathway​[20]​. The order of reaction steps and the fusion of riboflavin biosynthesis proteins (RIB) have undergone slight evolutionary alterations in distinct lineages​[21]​.\u003c/p\u003e \u003cp\u003eAnimals and plants also suffer severe repercussions when riboflavin is deficient, in plants weakened stress resistance and flawed seed development have been observed​[22\u0026ndash;24​]. Although the production and metabolism of riboflavin have been extensively studied, its physiological and biological applications are particularly limited in vegetables. In our best knowledge there have been limited study found about role of riboflavin in vegetables. In this piece of work, the potential benefits of riboflavin foliar treatment in alleviating drought stress in \u003cem\u003eMomordica charantia\u003c/em\u003e L. have been investigated.\u003c/p\u003e"},{"header":"Materials and Methodology","content":"\u003cp\u003eOn bitter gourd, experiment was carried out at the IMBB greenhouse at The University of Lahore, Lahore, to ascertain the functions of riboflavin (0, 25, 50, and 100 ppm) in reducing drought stress (100% and 60% FC). From the Ayub Agriculture Research Institute (AARI) in Faisalabad, bitter gourd seeds were gathered. Five seeds of bittergourd were planted in pots with 7 kg of thoroughly cleansed loamy soil.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePlant growth and fruit attributes\u003c/h2\u003e \u003cp\u003eManual measurements of the length of the shoot and the root were made using a scale. Fresh weight of the shoot, the root, and the fruit were also taken along with fruit and leaves count, leaf area, fruit diameter and volume of each replication. To record the data for the dry weights of the shoot and root, samples were oven-dried separately at 60\u0026deg;C until the stable mass was reached.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eDetermination of photosynthetic contents\u003c/h3\u003e\n\u003cp\u003eFollowing Arnon's method (1949)​[25]​, the contents of chlorophyll (\u003cem\u003ea, b\u003c/em\u003e, and total) and carotenoids were determined. Freshly harvested shoots(5g) were crushed in 10 ml of acetone (80%). A spectrophotometer was used to measure the filtrate's absorbance at 480, 645, and 663 nm, separately, after the filtrate was collected.\u003c/p\u003e\n\u003ch3\u003eDetermination of metabolites\u003c/h3\u003e\n\u003cp\u003eThe total phenolic content was ascertained using the method of Lamuela​[26]​ with Folin\u0026ndash;Ciocalteau phenol reagent. The absorbance was measured at 750 nm. The total phenolic content was determined as milligrams per gram of fresh leaf. The concentration of protein in the leaf extracts was measured following the method of Bradford (1976)​[27]​. About 2 ml of Bradford reagent and 0.1 ml of the leaf extracts were taken into test tubes and the mixture kept at room temperature for 5 min. The absorbance of each sample was measured at 595 nm. The method of Hamilton and Van Slyke (1943)​[28​] was followed to measure the total free amino acids. Results were found by mixing 10% pyridine and 1% ninhydrin with 1 ml of leaf extract, keeping the test tubes at room temperature for 30 minutes, and construing the optical density at 570 nm.\u003c/p\u003e\n\u003ch3\u003eDetermination of antioxidant enzyme activities\u003c/h3\u003e\n\u003cp\u003eFresh shoots (0.5 g) were crushed in buffer (50 mM, pH 7.8) on an ice bath. The mixture was then centrifuged for 20 minutes at 4\u0026deg;C using 15,000 rpm. The activity of antioxidant enzymes was assessed using the obtained supernatants. To measure the catalase (CAT) and peroxidase (POD) activities in bitterguord, the method of Chance and Maehly (1955)​[29​] was used. After 20 seconds, fluctuations in absorbance at 470 nm were observed for 3 minutes. A change in absorbance of 0.01 U/min was considered to be one unit of enzymatic activity. The DPPH activity was measured using the technique described by Valko​[30]​. After an incubation of the samples for 30min in dark, the absorbance was measured at 517nm wavelength. Enzyme activity was determined on the basis of soluble protein.\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eWith the aid of computer-based software (Statistix version 8.1), analysis of variance (ANOVA) was performed for each parameter. The data was represented graphically using Microsoft Excel.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eRiboflavin foliar application enhanced growth indices in bitter gourd\u003c/h2\u003e \u003cp\u003eStatistical analysis showed significant differences in plant height as well as root and shoot fresh and dry biomass (Table. 1). Drought reduced the root length by 27%, shoot length by 22%, root fresh and dry biomass by 31.2% and 53% while shoot fresh and dry biomass displayed a significant drop by 19.25% and 17% respectively, as compared to fully irrigated plants. Drought conditions also dropped number of leaves by 15% and leaf area by 20%. Riboflavin application under drought displayed positive effect on bitter gourd growth and biomass at all applied concentrations as plants exhibited a significant incline in root length by 18% at 25 ppm, 33% at 50 ppm and 40% at 100 ppm riboflavin concentration as compared to only drought treated plants. Similarly, bitter gourd shoot displayed improved growth by 12.8%, 25.4% and 35% at 25 ppm, 50 ppm and 100 ppm concentrations of riboflavin respectively, as compared to drought treated plants. Plant fresh and dry biomass also followed the similar trend and highest increase in root fresh and dry biomass was calculated at 100 ppm riboflavin by 63\u0026ndash;66% while shoot fresh and dry biomass was improved by 40\u0026ndash;42% as compared to only drought treated plants. Further, bitter gourd plants exhibited a significant incline in number of leaves by 14% at 25 ppm, 29% at 50 ppm and 37% at 100 ppm as well as leaf area increased by 17% at 100 ppm riboflavin concentration as compared to only drought treated plants. As shown in Figure \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e, the effect of riboflavin was significant on plant growth.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eRiboflavin foliar spray improved fruit attributes in bitter gourd\u003c/h3\u003e\n\u003cp\u003eSignificant differences were observed in yield attributes such as fruit count, fruit weight, fruit volume and fruit diameter (Table. 2). Drought reduced the fruit weight by 20%, number of fruits by 44%, fruit diameter by 22%, fruit volume by 43% as compared to fully irrigated plants. Riboflavin application under both control and drought conditions displayed positive effect on bitter gourd growth at all applied concentrations. Similarly, bitter gourd fruit weight displayed improved growth by 20%, 39.6% and 51.48% at 25 ppm, 50 ppm and 100 ppm concentrations of riboflavin respectively, as compared to drought treated plants. Number of fruit also followed the similar trend and highest increase in number of fruit was calculated at 100 ppm riboflavin by 28\u0026ndash;34% while fruit diameter and fruit volume was improved by 40% and 42% calculated at 100 ppm concentration of riboflavin respectively as compared to only drought treated plants. Riboflavin foliar spray significantly changed fruit count, size and weight properties.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eRiboflavin foliar spray enhanced photosynthetic contents in bitter gourd\u003c/h2\u003e \u003cp\u003eChlorophyll \u003cem\u003ea\u003c/em\u003e, chlorophyll \u003cem\u003eb\u003c/em\u003e, total chlorophyll and carotenoids showed positive responses with the increase of riboflavin as determined by statistical analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Drought reduced the chlorophyll \u003cem\u003ea\u003c/em\u003e by 24%, chlorophyll \u003cem\u003eb\u003c/em\u003e by 35%, total chlorophyll by 28%, and carotenoid by 24.35%, as compared to fully irrigated plants. Riboflavin foliar spray under drought displayed positive effect on bitter gourd growth at all applied concentrations as plants exhibited a significant incline in chlorophyll \u003cem\u003ea\u003c/em\u003e by 25% at 25 ppm, 26% at 50 ppm and 29% at 100 ppm riboflavin concentration as compared to only drought treated plants. Similarly, bitter gourd displayed increase contents of chlorophyll \u003cem\u003eb\u003c/em\u003e by 22.44%, 43% and 48% at 25 ppm, 50 ppm and 100 ppm concentrations of riboflavin respectively, as compared to drought treated plants. Total chlorophyll also followed the similar trend and highest increase was calculated at 100 ppm riboflavin by 30\u0026ndash;34% while carotenoids 16% was calculated at 100 ppm concentration of riboflavin respectively as compared to only drought treated plants. Riboflavin application under control conditions also exhibited positive effects on all above mentioned parameters. These results suggest, positive impact of riboflavin on plant pigments.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eRiboflavin foliar spray concerted regulation of metabolites in bitter gourd\u003c/h2\u003e \u003cp\u003ePlant metabolites such as total free amino acids, total soluble proteins and total phenolics showed positive responses with the increase of riboflavin concentration both under control and drought conditions (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Drought reduced the total free amino acids by 36.70% and total soluble protein by 30.25% respectively, as compared to fully irrigated plants. Riboflavin application under drought displayed positive effect on bitter gourd metabolites at all applied concentrations as plants exhibited a significant incline in total free amino acids by 30% at 25 ppm, 33% at 50 ppm and 41% at 100 ppm riboflavin concentration as compared to only drought treated plants. Similarly, bitter gourd total soluble proteins displayed improved contents by 15%, 28% and 37% at 25 ppm, 50 ppm and 100 ppm concentrations of riboflavin respectively, as compared to drought treated plants. Furthermore, drought increased the total phenol content by 24%, as compared to fully irrigated plants while under riboflavin application, plants further exhibited a significant incline in total phenols by 19% at 25 ppm, 20% at 50 ppm and 23% at 100 ppm riboflavin concentration as compared to only drought treated plants.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eRiboflavin foliar spray enhanced antioxidant enzyme activities in bitter gourd\u003c/h2\u003e \u003cp\u003eCatalase, peroxidase and DPPH assay showed positive response with the increase of riboflavin concentrations under both control and drought conditions as mentioned by statistical analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Drought conditions increased the catalase activity by 24.28%, peroxidase activity by 27.68% and DPPH assay by 51% as compared to fully irrigated plants. Riboflavin application under drought exhibited a significant decline in catalase by 11% at 25 ppm, 22.11% at 50 ppm and 42% at 100 ppm riboflavin concentration as compared to only drought treated plants. Similarly, bitter gourd peroxidase displayed decrease in enzyme activities by 17%, 27.47% and 36.50% at 25 ppm, 50 ppm and 100 ppm concentrations of riboflavin respectively, as compared to drought treated plants. DPPH assay also followed the similar trend and highest increase was calculated at 100 ppm riboflavin by 49\u0026ndash;56% concentration of riboflavin respectively as compared to only drought treated plants. These results strongly suggest that riboflavin foliar application strengthen the defense system of bitter gourd.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003ePlants react to various stressors through a variety of biological, morphological, and molecular pathways, when dealt with stress​[31]​. Many organisms including plants that live in soil are stressed by drought. Since desiccation is accompanied by an excessive intracellular generation of reactive oxygen species, antioxidants are particularly important. One antioxidant that controls a plant's ability to withstand drought is riboflavin (vitamin B2)​[32​]. The main function of water-soluble vitamin B2 (riboflavin) in cell biology is related to its conversion into FMN and FAD, the cofactors of numerous dehydrogenases, oxidases, and reductases engaged in a wide range of biological activities, including energetic metabolism and chromatin remodeling​[33]​. The production of vegetables is significantly impacted by drought. Drought stress has an impact on a number of plant functions, including photosynthesis, transpiration, the transfer of resources from source to sink, etc. Stomata are closed and leaf area is decreased in times of water scarcity, which lowers photosynthetic and transpiration activity and reduces water use​[34​]. Additionally, it slows down the physiological, biochemical, and morphological processes that contribute to plant growth. The primary responses of plants to stress include stunted growth of roots and shoots, sluggish rate of germination, closure of stomata, reduction in seedling growth, and worsening of photosynthetic activity​[35​]. In the current study, water stress had a noticeable impact on the morphological characteristics of plants. When compared to plants produced in a drought-prone environment, these metrics are much higher in plants grown under normal conditions. The findings of the current work are consistent with the results of Talebi and colleagues​[36​], who reported similar findings about Gazania plants. In accordance with their findings, the application of riboflavin at various concentrations improved the vegetative development metrics. The foliar application of riboflavin is crucial for the metabolic, biochemical, and physiological processes of plants (such as nutrient absorption, water relations, photosynthesis, and stress signaling, etc.), which ultimately have a positive impact on plant growth, yield, and crop quality as well as vegetative and reproductive development​[37​].\u003c/p\u003e \u003cp\u003eABA, also known as the stress hormone, is released in the roots and then goes to the plant's above-ground sections, where it sends signals for defenses against drought stress. Recently, a number of molecular mechanisms have been unveiled to determine the effect of hormones on ABA signal transduction genes and transcriptional profiles of stress related genes​[38,39]​. In pursuit of water, roots begin penetrating soils farther. Fruit failure results from decreased plant activity, which causes flowers to set fruit poorly or not at all[34]. Similar findings were made when working on bittergourd affected by drought; the fruit count along with its morphological parameters decreased, while, the application of riboflavin enhanced the aforementioned characteristics. The future depends in finding ways to combat the water shortage and drought stress that would guarantee abundant vegetable crop production to feed a growing population​[34]​.\u003c/p\u003e \u003cp\u003eUnder conditions of water stress, plants are less able to absorb nitrogen from the soil, which results in a decrease in plant biomass and nitrogen-related chemical components like total free amino acids and sugar​[40]​. In the current study, riboflavin considerably outperformed plants that were not supplemented with it in terms of total soluble proteins and total free amino acids under water stress circumstances. These outcomes are consistent with those reported by Thompson and colleagues​[41]​, who applied riboflavin topically to \u003cem\u003eNicotiana tabacum\u003c/em\u003e and saw comparable outcomes. Riboflavin improved the manufacture of amino acids such proline, glycine, serine, leucine, and lycine in \u003cem\u003eVitis vinifera\u003c/em\u003e, according to a subsequent study by Cangi and his colleagues​[42​]. As a molecular chaperone, proline works.\u003c/p\u003e \u003cp\u003eThe effects of the drought on crop productivity and the viability of agriculture were undesirable. It has a significant impact on the morphological and biochemical processes in plants, which inhibit growth and change enzyme activity at the cellular level​[43]​.\u003c/p\u003e \u003cp\u003eUncontrolled free radicals, increased, and plants use enzymatic and non-enzymatic antioxidants to reduce oxidants and manage cellular homeostasis under oxidative stress situations​[44​]. Severe oxidative stress can result from ROS buildup in plants​[45​]. The enzymatic antioxidant enzymes work together to counteract the excessive ROS production in plants, defending the structures and functions of cellular constituents. Generally speaking, under various abiotic stress conditions, the activity of several antioxidant enzymes in plants rises​[46\u0026ndash;48]​. According to Heidari​[49]​, an increase in enzymatic antioxidant activity is correlated with an increase in stress tolerance. When compared to normal plants, the antioxidant activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), catalase (CAT), and peroxidase (POD) were found to be improved by drought in the current study. These results are in line with those of Akram and his colleagues​[50​] (2012), who reported that drought stress caused a significant buildup of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e in leaves and MDA increased the activity of SOD, CAT, and POD in sunflower plants. According to several studies, the antioxidants catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), glutathione peroxidase (GPX), and polyphenol oxidase (APX) are among those that can be activated by riboflavin treatment in \u003cem\u003eHibiscus sabdariffa\u003c/em\u003e L.​[51]\u003csup\u003e​\u003c/sup\u003e.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003ePlant research now prioritizes studying how to preserve food production and how plants can withstand drought. All types of crops, including field crops, vegetable crops, fruit crops, spices, and others, are negatively impacted by drought. By decreasing both leaf area and photosynthetic rate per unit leaf area, drought stress is known to limit photosynthesis. It is essential to study the antioxidant enzyme research to comprehend the molecular mechanisms of various stress responses in bittergourd in order to increase crop production in controlled and water-deficit environments with the administration of vitamin B2 (riboflavin). Consequently, to lessen the negative effects of drought- generated oxidative stress, the findings showed a distinct pattern of antioxidants, where stimulation of DPPH, POD, and CAT activity was observed to be more effective in riboflavin-treated plants than in stressed plants. Because ROS created under stressful conditions can seriously damage the mechanical assembly of the photosynthetic system. Therefore, the use of riboflavin improved the morphological, biochemical and antioxidant activities of bitterguord under the influence of water scarcity.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eReactive Oxygen Species (ROS);; Ascorbic Acid,\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e(ASC); Superoxide Dismutase, (SOD); Catalase, (CAT), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and peroxidase (POD), superoxide dismutase (SOD), glutathione peroxidase (GPX). polyphenol oxidase (APX).\u003c/p\u003e\n"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003eResearchers supporting the project (RSP2024R393) at King Saud University, Riyadh,\u0026nbsp;Saudi\u0026nbsp;Arabia.\u0026nbsp;.\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest:\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAll authors declare no conflict of interest\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors Contributions:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;[AR] conceived the study and design the experiment. [NA] added valuable suggestions during experiment and manuscript improvement. [ZUN] contributed various inputs during the manuscript preparation and supervision. [MYA], [TF] performed data analysis and design the manuscript. Validation, funding contributed by [VR]. \u0026nbsp;[MAES] and [MI]\u003csup\u003e\u0026nbsp;\u003c/sup\u003einvolved in drafting article,\u0026nbsp;revision, arranged the figures, tables, experiment validation and manuscript description.\u0026nbsp;[AAS] approved the final version to be published.\u0026nbsp;All the authors read and approved the finalized manuscript for publication.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNot\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eapplicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAll authors give permission to the publisher to publish this research work.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAll data is presented in this manuscript. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe authors declare no known competing financial interest.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical Trial number\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments:\u0026nbsp;\u003c/strong\u003eThe authors extend their appreciation to the Researchers Supporting Project Number (RSP2024R182) King Saud University, Riyadh, Saudi Arabia\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eMubashir, A. et al. Effect of foliar application of nano-nutrients solution on growth and biochemical attributes of tomato (Solanum lycopersicum) under drought stress. Front. 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Engineering plants for heavy metal stress tolerance. Rend Lincei. 29, 709\u0026ndash;723 (2018). \u003c/li\u003e\n\u003cli\u003eZaid, A. \u0026amp; Mohammad, F. Methyl jasmonate and nitrogen interact to alleviate cadmium stress in menthaarvensis by regulating physio-biochemical damages and ROS detoxification. J Plant Growth Regul. 37, 1331\u0026ndash;1348 (2018). \u003c/li\u003e\n\u003cli\u003eZaid, A. \u0026amp; Wani, S. H. Reactive Oxygen Species Generation, Scavenging and Signaling in Plant Defense Responses. Bioactive Molecules in Plant Defense. (Springer, ; pp. 111\u0026ndash;132, 2019). \u003c/li\u003e\n\u003cli\u003eZaid, A., Mohammad, F. \u0026amp; Fariduddin, Q. Plant growth regulators improve growth, photosynthesis, mineral nutrient and antioxidant system under cadmium stress in menthol mint (Menthaarvensis L.) Physiol Mol Biol Plants. 26, 25\u0026ndash;39 (2020). \u003c/li\u003e\n\u003cli\u003eHeidari, P., Amerian, M. R. \u0026amp; Barcaccia, G. Hormone profiles and antioxidant activity of cultivated and wild tomato seedlings under low-temperature stress. Agronomy. 11, 1\u0026ndash;16 (2021). \u003c/li\u003e\n\u003cli\u003eAkram, N. A., Ashraf, M. \u0026amp; Al-Qurainy, F. Aminolevulinic acid-induced changes in some key physiological attributes and activities of antioxidant enzymes in sunflower (Helianthus annuus L.) plants under saline regimes. Sci Hortic 142, 143\u0026ndash;148 (2012). \u003c/li\u003e\n\u003cli\u003eAbdellatif, Y. M. R. \u0026amp; Ibrahim, M. T. S. Non-enzymatic anti-oxidants potential in enhancing Hibiscus sabdariffa L. tolerance to oxidative stress. International Journal of Botany 14, 43\u0026ndash;58 (2018). \u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTABLE 1: The effect of foliar application of Riboflavin on growth indices of bitter gourd under drought stress.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"666\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCondition\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFoliar ppm\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRoot length (cm)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eShoot Length (cm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRoot fresh weight(g)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eShoot fresh weight(g)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRoot dry weight(g)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eShoot dry weight(g)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLeaf Count\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(n)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLeaf Area\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(cm\u003csup\u003e3\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eControl\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e13.6\u003csup\u003ede \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e220\u003csup\u003ed \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e2.50\u003csup\u003ee\u0026nbsp;\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e16.1\u003csup\u003ede\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e0.45\u003csup\u003eef\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e3.4\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e150\u003csup\u003ed \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e23.4\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e15.5\u003csup\u003ebcd\u0026nbsp;\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e255\u003csup\u003ec \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e3.95\u003csup\u003ec\u0026nbsp;\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e18.5\u003csup\u003ecd\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e0.84\u003csup\u003ec\u0026nbsp;\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e4.5\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e172\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e27.1\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e17.4\u003csup\u003eb \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e289\u003csup\u003eb \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e4.72\u003csup\u003eb\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e22.9\u003csup\u003eb\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e1.10\u003csup\u003eb\u0026nbsp;\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e4.9\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e197\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e32.2\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e19.4\u003csup\u003ea \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e325\u003csup\u003ea \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e5.9\u003csup\u003ea \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e26.5\u003csup\u003ea\u0026nbsp;\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e1.31\u003csup\u003ea\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e5.8\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e217\u003csup\u003ea \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e36.4\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDrought\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e9.8\u003csup\u003ef \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e170\u003csup\u003ef \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e1.72\u003csup\u003ef \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e13.0\u003csup\u003ee \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e0.21\u003csup\u003eg \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e2.8\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e122\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e17.1\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e12.0\u003csup\u003ee \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e195\u003csup\u003ee \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e2.44\u003csup\u003ee \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e15.9\u003csup\u003ede \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e0.35\u003csup\u003efg \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e3.4\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e147\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e21.3\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e14.7\u003csup\u003ecd \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e228\u003csup\u003ed \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e3.40\u003csup\u003ed \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e19.5\u003csup\u003ec \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e0.54\u003csup\u003ede \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e4.0\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e170\u003csup\u003ec \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e27.1\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e16.5\u003csup\u003eb \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e265\u003csup\u003ec \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e4.73\u003csup\u003eb \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e22.7\u003csup\u003eb \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e0.63\u003csup\u003ed \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e4.7\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e189\u003csup\u003eb \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e30.1\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 160px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eANOVA (F Value)\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDrought(D)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFoliar(F)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 110px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDrought*foliar \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003ens \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003ens \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e***\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003ens \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003e***\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 68px;\"\u003e\n \u003cp\u003ens \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 50px;\"\u003e\n \u003cp\u003ens \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003ens \u0026nbsp; \u0026nbsp;\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\u003eEach value is a mean of four replicates; different alphabetic letters indicate significant differences (P\u0026le;0.05) among treatments, *, ** and *** indicated significance at P\u0026le;0.05, P\u0026le;0.01 and P\u0026le;0.001 respectively; ns indicated non-significant differences. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTABLE 2: The effect of foliar application of Riboflavin on fruit attributes of bitter gourd under drought stress.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"498\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCondition\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFoliar ppm\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFruit Weight\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(g)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNumber of Fruits\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(n)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFruit Diameter\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFruit Volume\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eControl \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e13.6\u003csup\u003ed \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e6\u003csup\u003ecd\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e1.7\u003csup\u003ed\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e6.9\u003csup\u003ed\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e16.4\u003csup\u003ec \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e7\u003csup\u003ebc\u0026nbsp;\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e12.1\u003csup\u003ec\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e9.7\u003csup\u003ec\u0026nbsp;\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e21.1\u003csup\u003eb \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e8\u003csup\u003eb\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e2.5\u003csup\u003eb\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e12.2\u003csup\u003eb\u0026nbsp;\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026nbsp;\u003c/sup\u003e26.3\u003csup\u003ea \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026nbsp;\u003c/sup\u003e10.7\u003csup\u003ea \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026nbsp;\u003c/sup\u003e3.0\u003csup\u003ea\u0026nbsp;\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026nbsp;\u003c/sup\u003e15.3\u003csup\u003ea\u003c/sup\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDrought\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e10.1\u003csup\u003ee \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e2.2\u003csup\u003ef \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e1.3\u003csup\u003ef \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e3.8\u003csup\u003ef \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e13.9\u003csup\u003ed \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e4\u003csup\u003eef\u003cstrong\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/strong\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e1.6\u003csup\u003ee \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e5.5\u003csup\u003e\u0026nbsp;e \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e17.1\u003csup\u003ec \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e5\u003csup\u003ede \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e2.0\u003csup\u003ec \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e7.3\u003csup\u003ed\u003cstrong\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/strong\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e20.9\u003csup\u003eb \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e7\u003csup\u003ebc \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e2.4\u003csup\u003eb \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e8.8\u003csup\u003ec \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eANOVA (F Value)\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDrought(D)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFoliar(F)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDrought*foliar \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; ** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003ens \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e**\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e*** \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\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\u003eEach value is a mean of four replicates; different alphabetic letters indicate significant differences (P\u0026le;0.05) among treatments, *, ** and *** indicated significance at P\u0026le;0.05, P\u0026le;0.01 and P\u0026le;0.001 respectively; ns indicated non-significant differences.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\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":"","lastPublishedDoi":"10.21203/rs.3.rs-5356796/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5356796/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eDrought is an abiotic stress for many species that live in soil. Although many plants have been studied for drought-adaptive processes, but a little is known about Bittergourd (\u003cem\u003eMomordica charantia L.\u003c/em\u003e). Antioxidants are especially important because desiccation causes an increase in the intracellular generation of reactive oxygen species. Riboflavin (vitamin B2) is one antioxidant that influences plant drought tolerance. It causes plants to produce reactive oxygen species(ROS) when exposed to light, is an excellent photosensitizer for biocidal reactions. This study explores the possible protective role of riboflavin (0, 25, 50, 100 ppm) foliar application against drought stress (i.e., 60% and 100% field capacity) in bitter gourd plants. A pot experiment was conducted in a completely randomized design (CRD) with four replicates during spring season of 2021 at the field area of University of Lahore. Results indicated that the foliar applications of riboflavin (RF) improved morphological and physiological attributes in plants as compared to control plants when grown under drought. Although with increasing concentration of riboflavin all the parameters gradually increased. Fruit count (44%), weight (20%), diameter (22%) and volume (43%) also increased with foliar application. Photosynthetic pigments (Chlorophyll \u003cem\u003ea\u003c/em\u003e by 29%, Chlorophyll \u003cem\u003eb\u003c/em\u003e by 48%, Total chlorophyll by 34% and Carotenoid by 16%) and metabolites (Total free amino acids by 41% and Total soluble proteins by 37%) increased under the influence of riboflavin. Moreover, Phenolic contents and antioxidant activities (Catalase, Peroxidase and DPPH) of bitter gourd were highlighted showing significant increase in stress which further enhanced under the effect of riboflavin by 23%, 42%, 36% and 56%, respectively to determine the specific direction to be taken for future plant antioxidant research.\u003c/p\u003e","manuscriptTitle":"Riboflavin (Vitamin B2)-induced biomass, yield and antioxidant potential of Bittergourd in water-deficit condition","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-13 11:42:20","doi":"10.21203/rs.3.rs-5356796/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":"b715a696-685c-4a8e-8720-b3ba47c42675","owner":[],"postedDate":"November 13th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-05-07T16:08:25+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-13 11:42:20","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5356796","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5356796","identity":"rs-5356796","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}