Reaction of Luffa spp. to Macrophomina pseudophaseolina inoculation

Reaction of Luffa spp. to Macrophomina pseudophaseolina inoculation | 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 Reaction of Luffa spp. to Macrophomina pseudophaseolina inoculation Dariane Monteiro Viana, Andréia Mitsa Paiva Negreiros, Lindomar Maria Silveira, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4094029/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 The genus Macrophomina is responsible for causing diseases in various species of the Cucurbitaceae family. This study aimed to evaluate the reaction of Luffa species when associated with M. pseudophaseolina . Initially, a prospection study was carried out to identify fungi associated with Luffa roots. Three isolates belonging to the M. pseudophaseolina species were identified and confirmed by DNA extraction and PCR amplification, using specific primers. One isolate was selected to test pathogenicity on Luffa . Twelve Luffa spp. accessions from the cucurbitaceous germplasm collection of Universidade Federal Rural do Semi-Árido were used, along with two positive controls: melon and watermelon plants. Inoculation was performed using the infested toothpick method. The plants were maintained under greenhouse conditions in a completely randomized design with four replications. The trial was repeated. The following variables were assessed: disease incidence and severity, reaction class, shoot length (SL), root length (RL), fresh shoot weight (FSW), dry shoot weight (DSW), fresh root weight (FRW), and dry root weight (DRW). Accession AB39 proved to be immune to the pathogen, as well as exhibiting the highest values of SL (163.9 cm), RL (43.4 cm), FSW (24.9 g), FRW (9.8 g), DSW (3.4 g), and DRW (0.5 g). These results indicate that the immune resistance of AB39 makes this accession promising, both in terms of disease resistance and vegetative growth, standing out as the best option among the accessions evaluated. Additionally, this is the first report of Luffa spp. acting as an alternative host of M. pseudophaseolina . Loofah Cucurbitaceae Reaction of accessions Root pathogens Figures Figure 1 Figure 2 Figure 3 1 ǀ INTRODUCTION Macrophomina is a fungal genus belonging to the family Botryosphaeriaceae, class Dothideomycetes. The species Macrophomina pseudophaseolina Crous, Sarr & Ndiaye, was first reported in Brazil in 2018, causing charcoal rot on oilseeds and jatropha seeds. More recently, it has been found on asymptomatic roots of weeds present in melon and watermelon production fields in the Northeast region of Brazil, as well as on cassava branches in Minas Gerais (Machado et al., 2018 ; Brito et al., 2019 ; Negreiros et al., 2019 ). Macrophomina phaseolina (Tassi) G. Goid. and M. pseudophaseolina are reported to be the most prevalent species in Brazil, both of which were considered pathogenic to melon trees in pathogenicity tests (Negreiros et al., 2019 ; Negreiros et al., 2022 ). The symptoms of Macrophomina spp. generally include watery, light brown lesions with droplets of translucent exudate, which later darken, dry out, and become whitish, with longitudinal cracks. In severe cases, premature plant death occurs due to the blockage of the vascular system (xylem) caused by hyphal colonization, preventing the passage of water and nutrients to the plant (Islam et al., 2012 ; Pereira et al., 2012 ; Marquez et al., 2021 ). This genus exhibits good adaptation to regions with arid and semi-arid climates, thriving at temperatures between 26 and 38°C. It spreads through microsclerotia in the host's tissues or cultural remains as pycnidia, which can be observed as black dots on infected tissues (Almeida et al., 2008 ; Gupta et al., 2012 ; Pereira et al., 2012 ; Sarr et al., 2014 ; Negreiros et al., 2022 ). Currently, there are no registrations of synthetic active ingredients (fungicides) for the control of these phytopathogens in melon and watermelon production areas in Brazil. However, biological products based on Trichoderma spp. and Bacillus spp. are registered for the control of M. phaseolina in other crops (Agrofit, 2024). Nevertheless, the risks to the soil's micro and macrobiota from using these products are being investigated. Given this scenario, the most recommended approach involves managing cultural remains, using treated seeds, employing resistant genotypes, and utilizing rootstocks (Cohen et al., 2022 ; Pereira et al., 2012 ; Sales Júnior et al., 2019 ). The use of resistant rootstock is an effective strategy that enables plants to complete their life cycle in the field and is considered a less invasive solution for dynamic soil ecosystems (Jang et al., 2014 ; Cohen et al., 2022 ). However, for the grafting technique to be effective, it is necessary to select plant material that is compatible with the target crop, easy to replicate, and provides resistance to these pathogens (Davis et al., 2008 ). According to Li et al. ( 2016 ) and Bindal et al. ( 2023 ), Luffa spp. seedlings can serve as rootstocks for other cucurbits such as pumpkin, cucumber, and melon. Hilal et al. ( 2000 ) tested fungi isolated from the roots and seeds of Luffa aegyptiaca Mill and found that M. phaseolina was not pathogenic to this crop. Subsequent studies also highlighted the resistance of this cucurbitaceous plant when inoculated with other root pathogens (Bruton et al., 2000 ; Kwon et al., 2001 ; Namisy et al., 2023 ; Bindal et al., 2023 ). The lack of knowledge about the influence of diseases transmitted by root pathogens has hindered the exploitation of this material for possible agricultural purposes (Bindal et al., 2023 ). Further research is therefore needed to better understand these effects and develop appropriate management strategies. In this context, it is important to emphasize that the Luffa genus shows variability in various characteristics and has a considerable number of resistance genes (Wu et al., 2020 ). It should also be noted that some species of this genus, such as Luffa cylindrica M. Roem, are considered allogamous, reproducing preferentially by crossing. Therefore, evaluations are necessary to determine the resistance of these species to be used as rootstocks resistant to soil pathogens. Given the above, this study aimed to assess the pathogenicity of M. pseudophaseolina on Luffa spp. accessions to identify resistant materials that can later be used in breeding programs and/or directly as rootstocks for other cucurbits. 2 ǀ MATERIALS AND METHODS 2.1 Obtaining and characterizing fungi from the roots of Luffa spp. A prospecting study of fungi associated with roots of Luffa spp. was conducted between February and May 2022 in the states of Ceará (CE) and Rio Grande do Norte (RN). Nineteen root samples were collected from asymptomatic plants. Plant roots were washed in running water to remove soil residues, followed by immersion in a 1% sodium hypochlorite (NaClO) solution for one minute and rinsing with sterilized water to remove excess chlorine. Seven root fragments were placed in Petri dishes containing potato-dextrose-agar (PDA) medium with 0.5 g.L -1 of tetracycline added (Mello et al., 2011 ). The plates containing root fragments were incubated in a Biochemical Oxygen Demand (BOD) incubator at 28 ± 1°C in the dark for up to seven days. Fungal colonies growing on root fragments were transferred to Petri dishes containing PDA medium. Fungal isolates were identified to the genus level by preparing slides containing the fungal structures stained with lactophenol cotton blue for observation under an optical microscope and compared with descriptions in fungal identification keys (Barnett & Hunter, 1998 ; Seifert et al., 2011 ). The monohyphal purification technique was used to obtain pure cultures, which were then preserved using the Castellani method (1939). The isolates obtained were deposited in the fungal culture collection of the Phytopathology Laboratory II at Universidade Federal Rural do Semi-Árido (UFERSA). For the molecular characterization of Macrophomina species, the polymerase chain reaction (PCR) technique with specific primers was used. Fungal isolates were cultivated in PDA medium with cellophane overlaid to allow the mycelia to be obtained, and subsequently the deoxyribonucleic acid (DNA) of each isolate to be extracted. Genomic DNA was extracted using the Wizard ® genomic DNA purification kit (Promega Corporation, Madison, USA), following the manufacturer's protocol. PCR amplification was carried out using 6.25 µL of GoTaq ® Green Master Mix (2X) from Promega Corporation (Madison, USA), 4.25 µL of Milli-Q ® water, 0.5 µl of each primer (forward and reverse), and 1 µl of DNA (25 ng/µL). Water was used for the negative control instead of DNA, while isolates previously identified with specific primers were used for the positive control. Specific primers from the translation elongation factor-1alpha (TEF1-α) locus were used for M. phaseolina (MpTefF-AAACACACTTTTCGCACTCCTGC, MpTefR-TATGCTCGCAGAGAAGAACACGA), M. pseudophaseolina (MsTefF-GCACACTTTTCGCGCTTCTGTA, MsTefR-TGTGCTCGCTGGGAAGAACATGA), and M. euphorbiicola (MeTefF-AAGCATACTTTTCGTGCTCCTGC, MeTefR-AAAGGAACATGAGTGGCCAAAAA) (Santos et al., 2020 ). Thermal cycler was used with the following program: initial denaturation at 94°C for two minutes, 30 cycles of denaturation at 94°C for one minute, annealing at 63°C for 30 seconds, extension at 72°C for one minute, and a final extension at 72°C for ten minutes. After completing the amplification cycles, PCR products were separated by gel electrophoresis, loading 5 µL of the amplified DNA together with 3 µL of sample buffer + Gel Red 2X (Santos et al., 2020 ). Finally, results were visualized under ultraviolet (UV) light. The species of the pathogen was confirmed by amplifying the products using primers specific to target species. All molecular techniques used to identify isolates were conducted in the Laboratory of Molecular Biology at the Universidade Federal do Cariri (UFCA), in the state of CE, Brazil. For the subsequent virulence test, a Macrophomina isolate was selected, identified as M. pseudophaseolina (RO-II), based on preliminary pathogenicity evaluations showing greater aggressiveness on L. cylindrica and 'Natal' melon plants (unpublished data). 2.2 Luffa spp. accessions used to evaluate resistance to M. pseudophaseolina Twelve Luffa spp. accessions from the Cucurbitaceae Germplasm Collection of the Department of Agronomic and Forestry Sciences of the Centre for Agrarian Sciences at UFERSA were used (Table 1 ). Table 1 Accessions of Luffa spp. used in the experiment Accession code a Identification Year of collection Place of collection b AB07 Luffa cylindrica 2010 Apodi/RN AB24 Luffa sp. 2014 Petrolina/PE AB26 Luffa cylindrica 2010 Apodi/RN AB36 Luffa cylindrica - Serra Talhada/PE AB39 Luffa cylindrica 2012 Petrolina/PE AB40 Luffa cylindrica 2012 Gov. Dix-Sept Rosado/RN AB41 Luffa cylindrica 2012 São José do Belmonte/PE AB44 Luffa cylindrica 2012 Apodi/RN AB50 Luffa cylindrica 2013 Orocó/PE AB51 Luffa cylindrica 2013 Monsenhor Tabosa/CE AB52 Luffa cylindrica 2014 Serra Talhada/PE AB55 Luffa cylindrica 2014 Serra Talhada/PE a Code of accessions in the Cucurbitaceae Germplasm Collection. b RN = Rio Grande do Norte State; PE = Pernambuco State; CE = Ceará State. 2.3 Setting up and conducting the experiment To assess the reaction of Luffa species to RO-II, a test was conducted in a greenhouse at UFERSA's West Campus in the municipality of Mossoró (RN), following a completely randomized design (CRD). Twelve accessions of Luffa spp., yellow melon cultivar 'Natal', and watermelon cultivar 'Crimson Sweet' were inoculated with the fungal isolate, along with negative controls for each accession/cultivar. The experiment was repeated. Luffa spp. seeds underwent disinfection and dormancy overcoming processes (Oliveira et al., 2012 ; Medeiros et al., 2019 ). Seedlings were sown in polythene trays containing commercial substrate Tropstrato HT Hortaliças ® (Vida Verde, Brazil) and watered daily. After showing the first definitive leaf, seedlings were transferred to 1 L pots containing a mixture of sand and autoclaved commercial substrate in a 3:1 ratio (v/v). Irrigation was carried out daily until the soil reached field capacity. Ten days after sowing, the M. pseudophaseolina isolate was inoculated using the method described by Ambrósio et al. ( 2015 ), which involved inserting toothpicks colonized with the isolate's mycelium. Uninfested and autoclaved toothpicks were used as negative controls. The trial was evaluated 60 days after seedling transplantation. Plants were gently removed from pots, and root systems were carefully washed with running water. The variables analyzed included disease incidence and severity, average plant reaction to the pathogen, shoot and root lengths (SL and RL), fresh shoot and fresh root weights (FSW and FRW), and dry shoot and root weights (DSW and DRW). Disease incidence was determined as the number of infected plants per treatment and expressed as percentage (%), while its severity was assessed by a diagrammatic grading scale. For M. pseudophaseolina -inoculated plants, we used a modified scale described by Ravf & Ahmad ( 1998 ), with scores ranging from 0 to 5, wherein: 0 = no symptoms; 1 = less than 3% of infected tissues; 2 = 3–10% of infected tissues; 3 = 11–25% of infected tissues; 4 = 25–50% of infected tissues; and 5 = more than 50% of infected tissues. To assess the overall reaction of plants to disease, we employed a scale outlined by Salari et al. ( 2012 ). This scale calculates the average score per treatment by summing the scores assigned to each individual plant, dividing by the total number of plants evaluated, and subsequently categorizing the results as follows: 0 = immune (I); 0.1-1.0 = highly resistant (HR); 1.1-2.0 = moderately resistant (MR); 2.1-4.0 = susceptible (SU); and 4.1-5.0 = highly susceptible (HS). Shoot and root length were measured using a tape measure (cm). Fresh and dry weights of shoots and roots were determined using an analytical balance (g). To obtain the dry weights, each plant was placed individually in paper bags and subjected to a forced air circulation oven at 70°C until a consistent dry weight was achieved. 2.4 Statistical analysis Incidence and severity results were analyzed using the non-parametric Kruskal-Wallis test at a 5% probability level. Length, fresh and dry weights of shoots and roots were analyzed using ANOVA, and means were compared using the Scott-Knott test at a 5% probability level. Statistical analyses were conducted using Assistat 7.7 software (Silva & Azevedo, 2016 ). 3 ǀ RESULTS AND DISCUSSION 3.1 Isolation and characterization of root-associated fungi in Luffa spp. From asymptomatic Luffa spp. roots, nineteen fungal isolates were obtained. Using an identification key, we successfully classified eight of these isolates to the genus level, namely: Lasiodiplodia sp. (5.0%), Chaetomium sp. (5.0%), Rhizoctonia sp. (16.0%), and Macrophomina sp. (16.0%) (Fig. 1 ). However, conventional morphological identification methods proved inadequate for the remaining eleven isolates. Further characterization of the three Macrophomina isolates identified through morphological features confirmed them as M. pseudophaseolina using PCR analysis (Table 2 ). Table 2 Fungal genera collected from Luffa spp. roots in the states of Ceará (CE) and Rio Grande do Norte (RN) Identification code a Isolates Location Geographical coordinates RO – II Macrophomina pseudophaseolina Brazil, Rio Grande do Norte, Mossoró 5°12'23.26"S/ 37°19'45.70"W RO – III Macrophomina pseudophaseolina Brazil, Rio Grande do Norte, Mossoró 5°12'24.91"S/ 37°19'46.29"W QZ – I Macrophomina pseudophaseolina Brazil, Rio Grande do Norte, Mossoró 5°12'30.05"S/ 37°20'11.99"W OV – I Rhizoctonia sp. Brazil, Rio Grande do Norte, Mossoró 5°12'25.92"S/ 37°18'59.85"W BM Rhizoctonia sp. Brazil, Rio Grande do Norte, Mossoró 5°12'20.23"S/ 37°19'07.46"W BR- I Rhizoctonia sp. Brazil, Rio Grande do Norte, Mossoró 5°13'39.38"S/ 37°19'17.17"W CA – II Lasiodiplodia sp. Brazil, Rio Grande do Norte, Mossoró 5°11'48.95"S/ 37e19'14.38"W R5 – II Chaetomium sp. Brazil, Ceará, Caucaia 3°43'55.13"S/ 38°38'41.69"W a Isolates deposited in the collection of fungal isolates at the Phytopathology Laboratory II of the Universidade Federal Rural do Semi-Árido (UFERSA). 3.2 Response of Luffa spp. accessions to M. pseudophaseolina inoculation Data from both experiments were combined since no significant effect of the experiment repetitions was observed (ANOVA p > 0.05) for all the variables analyzed. Inoculation of cucurbits by M. pseudophaseolina revealed statistically significant differences for disease incidence and severity, as demonstrated by the Kruskal-Wallis test at a 5% probability ( p ≤ 0.05) (Table 3 ), and for SL, RL, FSW, FRW, DSW, and DRW, as indicated by the Scott-Knott test at a 5% probability ( p ≤ 0.05) (Table 4 ). Table 3 Disease incidence and severity, and reaction class against Macrophomina pseudophaseolina inoculation in accessions of Luffa spp., Cucumis melo , and Citrullus lanatus . Treatment/ accessions Disease incidence Disease severity Reaction c Rank a Mean (%) Rank a Mean b AB07 67.5 b 100.0 45.1 a-d 2.1 SU AB24 46.5 ab 63.0 37.1 a-c 1.7 MR AB26 67.5 b 100.0 60.2 a-d 3.1 SU AB36 67.5 b 100.0 60.2 a-d 3.1 SU AB39 11.5 a 0.0 11.5 a 0.0 I AB40 67.5 b 100.0 85.4 cd 4.5 HS AB41 67.5 b 100.0 93.0 d 5.0 HS AB44 67.5 b 100.0 85.0 cd 4.6 HS AB50 67.5 b 100.0 93.0 d 5.0 HS AB51 39.5 ab 50.0 32.5 a-c 1.5 MR AB52 67.5 b 100.0 65.0 a-d 3.5 SU AB55 53.5 b 75.0 27.2 ab 0.8 HR Melon 67.5 b 100.0 75.4 b-d 4.1 HS Watermelon 32.5 ab 38.0 21.1 ab 0.5 HR χ 2 65.5 80.3 χ 2 = significant chi-squared values at 5% probability. a Values followed by the same letter within columns do not differ statistically from each other by the Kruskal-Wallis non-parametric test ( p ≤ 0.05). b Average scores for all observations within each sample following the diagrammatic scale: 0 (no symptoms) and 5 (more than 50% of infected tissues) (Ravf & Ahmad, 1998 ). c Reaction to the disease according to Salari et al. ( 2012 ), wherein: I = immune; HR = highly resistant; MR = moderately resistant; SU = susceptible; and HS = highly susceptible. Data are average values from two experiments, each with four replicates (pots) per treatment and one plant per pot. Table 4 Averages of lengths and fresh and dry weights of shoots and roots in accessions of Luffa spp., Cucumis melo , and Citrullus lanatus inoculated with Macrophomina pseudophaseolina . Treatment/accessions SL a (cm) RL b (cm) FSW c (g) FRW d (g) DSW e (g) DRW f (g) AB07 111.5 b 25.5 c 20.0 b 6.7 b 2.8 a 0.4 b AB24 139.5 a 29.4 c 14.7 c 6.2 b 2.6 a 0.3 c AB26 112.9 b 25.7 c 10.7 d 4.0 d 2.1 a 0.2 d AB36 122.0 b 26.0 c 16.6 c 6.0 b 2.5 a 0.3 c AB39 163.9 a 43.4 a 24.9 a 9.8 a 3.4 a 0.5 a AB40 58.5 c 18.1 d 4.2 e 2.0 e 1.2 b 0.2 d AB41 47.0 c 16.3 d 6.5 e 0.5 f 1.3 b 0.1 e AB44 69.1 c 16.0 d 6.7 e 1.5 e 1.4 b 0.1 e AB50 63.0 c 24.5 c 5.2 e 2.3 e 1.2 b 0.2 d AB51 156.5 a 35.2 b 21.4 b 5.2 c 3.0 a 0.3 c AB52 115.2 b 23.0 c 12.1 d 5.2 c 2.5 a 0.3 c AB55 113.2 b 24.0 c 16.0 c 4.7 c 2.0 b 0.3 c Melon 46.2 c 21.0 d 15.0 c 2.3 e 1.5 b 0.1 e Watermelon 83.0 c 26.6 c 22.5 b 2.6 e 3.0 a 0.2 d CV (%) 26.99 30.45 23.48 22.21 20.0 18.17 CV (%) = significant coefficients of variation; values followed by the same letter within columns do not differ statistically from each other by the Scott-Knott test ( p ≤ 0.05). a Shoot length. b Root length. c Fresh shoot weight. d Fresh root weight. e Dry shoot weight. f Dry root weight. For disease incidence, treatments AB24 (63.0%), AB39 (0.0%), and AB51 (50.0%) showed no significant difference compared to Watermelon (38.0%). Notably, among the Melon treatments, only AB39 (0.0%) exhibited a statistically different result remained unaffected by the pathogen, indicating its immunity to the disease (Table 3 ). The reaction to M. pseudophaseolina revealed that nine out of the 12 Luffa spp. accessions exhibited a disease incidence of over 75% (AB07, AB26, AB36, AB40, AB41, AB44, AB50, AB52, and AB55) (Table 3 ). Remarkably, only accession AB39 remained unaffected by the disease. Comparatively, among the positive controls, Watermelon showed the least susceptibility. This aligns with findings by Negreiros et al. ( 2022 ), who observed higher disease incidence on melon (40% and 80% for isolates CMM4771 and CMM4801, respectively) compared to watermelon (0% and 40%, respectively), supporting the results of this study. Regarding disease severity, Luffa spp. accessions AB40 (4.5), AB41 (5.0), AB44 (4.6), and AB50 (5.0) differed from Watermelon (0.5). However, when comparing treatments inoculated with M. pseudophaseolina to Melon (4.1), only AB39 (0.0) showed a statistically significant difference (Table 3 ). Notably, the infection by M. pseudophaseolina in accessions AB41, AB44, and AB50 initiated around the 19th day after inoculation. Various structures such as microsclerotia and pycnidia were observed in close proximity to the plant's neck. Additionally, symptoms such as cracks in the hypocotyl and chlorosis on lower leaves appeared, ultimately resulting in the death of the plants (Fig. 2 ). Based on the reaction classification proposed by Salari et al. ( 2012 ), AB39 demonstrated immunity to the pathogen, while AB55 exhibited a highly resistant reaction, akin to the positive control Watermelon. Treatments AB24 and AB51 were categorized as moderately resistant, whereas treatments AB07, AB26, AB36, and AB52 were deemed susceptible to the pathogen. Treatments AB40, AB41, AB44, AB50, and Melon were classified as highly susceptible (Table 3 ) (Fig. 3 ). Hilal et al. ( 2000 ) conducted inoculation trials on L. aegyptiaca plants infected with M. phaseolina and reported the absence of symptoms. However, when combined with another pathogen, Pythium sp., visible symptoms emerged. Interestingly, the authors noted that the same M. phaseolina isolate (derived from L. aegyptiaca ) exhibited high pathogenicity when introduced to various other crops, including cotton 'Giza 75', cucumber 'Amira 2', okra 'Costa Dourada', peanut 'Giza 4', pumpkin 'Balady', sesame 'Giza 32', soya 'Crawford', zucchini 'Eskandarani', and watermelon 'Giza 1'. Wu et al. ( 2020 ) noted that the significant genetic variability within Luffa spp. may result in certain accessions possessing enriched resistance genes. These genes enhance the plants' capacity to withstand various environmental stresses, including water scarcity, temperature fluctuations, high humidity, and diseases. Namisy et al. ( 2023 ) observed similar findings, noting variations in the reactions of different accessions when inoculated with Fusarium oxysporum f. sp. luffae . The authors attributed these differences to the influence of plant genotype on disease susceptibility and resistance. For the biometric variable SL, treatments AB24, AB39, and AB51 demonstrated statistically superior performance compared to other treatments, with average values ranging from 139.5 to 163.9 cm, representing the highest recorded lengths. Conversely, treatments AB07, AB26, AB36, AB52, and AB55 significantly differed from both Melon and Watermelon, displaying lengths ranging from 111.5 to 122.0 cm. However, treatments AB40, AB41, AB44, and AB50 (ranging from 47.0 to 69.1 cm) did not exhibit significant differences compared to Melon and Watermelon (measuring 46.2 and 83.0 cm, respectively), indicating the shortest lengths. Accession AB39 stood out statistically among all treatments for the variable RL (43.4 cm), FSW (24.9 g), FRW (9.8 g), and DRW (0.5 g), boasting the highest average values across these variables. Conversely, for the RL variable, treatments AB40, AB41, AB44, and Melon showed significantly lower averages (ranging from 16.0 to 21.0 cm), indicating a notable reduction in this characteristic. Similarly, in terms of FSW, treatments AB40, AB41, AB44, and AB50 exhibited inferior values compared to other treatments, with averages ranging from 4.2 to 6.7 g. Regarding FRW, AB41 displayed the lowest average value (0.5 g) among treatments, indicating a greater reduction in the root system. With respect to DRW, the lowest recorded weight was 0.1 g for treatments TB41, TB44, and Melon. Concerning DSW, treatments AB07, AB24, AB26, AB36, AB39, AB51, AB52, and Watermelon demonstrated statistically superior performance, ranging from 2.1 to 3.4 g. Conversely, treatments AB40, AB41, AB44, AB50, and AB55 exhibited average values ranging from 1.2 to 2.0 g, which differed significantly from the Watermelon treatment but not from the Melon treatment. As Macrophomina spp. is a fungus naturally present in the soil and associated with its chemical and physical conditions, besides constantly interacting with other pathosystems, any strategy for disease management should aim to minimize alterations or manipulations of environmental and natural resources (Marquez et al., 2021 ). In this sense, pathogenicity studies play a crucial role in identifying resistant plant materials, as controlling Macrophomina spp. is inherently complex. Therefore, it is vital to implement one or a combination of control methods to mitigate the detrimental effects of phytopathogen attacks on vulnerable crops, thereby reducing inoculum pressure and the incidence of disease (Agrofit, 2024). Grafting stands out as one of the most effective and environmentally friendly methods for controlling Macrophomina spp. in agricultural fields. Cohen et al. ( 2022 ) emphasized its success in select regions of Israel, where it has effectively managed this pathogen in melon and watermelon crops by utilizing pumpkin rootstock resistant to the disease. However, this approach necessitates careful selection of plant materials that not only align with target crops but also offer replicability and resistance to pathogens (Davis et al., 2008 ). In recent years, researchers have explored Luffa species as potential rootstocks for cucurbits such as pumpkin, cucumber, and melon (Li et al., 2016 ; Shahiba & Chacko, 2019; Nordey et al., 2020 ; Guo et al., 2023 ), making accession AB39 particularly promising for breeding programs aimed at developing resistant cultivars. Moreover, it is noteworthy that this study marks the first documentation of Luffa spp. acting as an alternative host for M. pseudophaseolina , thus contributing significantly to our understanding of the interaction between M. pseudophaseolina and Luffa species. These findings offer valuable insights for managing the disease in other cucurbit crops. Additionally, accession AB39 exhibited the highest averages for RL and FRW among all treatments. As highlighted by Katuuramu et al. ( 2020 ), robust root development enhances nutrient and water absorption while mitigating severity of diseases transmitted by soil pathogens. 4 ǀ CONCLUSION Accession AB39 was classified as "immune" to the pathogen. This is the first report of Luffa spp. acting as an alternative host for Macrophomina pseudophaseolina . Declarations Acknowledgments This study was funded by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) – Brazil. Conflict of interest statement The authors of the manuscript declare that there are no personal, commercial, academic, political, or financial conflicts of interest in the manuscript. Authors' contributions AMPN, LMS, and RSJ contributed to the conception and design of the study. DMV, AMPN, ALAC, and CPSSA set up the tests and collected data. DMV and AMPN performed the statistical analyses. DMV, AMPN, LMS, ALAC, and RSJ assisted with the writing, proofreading, and editing of the manuscript. All authors contributed to the data analysis, interpretation of the results, and writing of the manuscript. References AGROFIT. Sistemas de agrotóxicos fitossanitários . Available at: . Accessed on: Jan 23, 2024. Almeida, A. M. R., Sosa-Gomez, D. R., Binneck, E., Marin, S. R. R., Zucchi, M. I., Abdelnoor, R. V., & Souto, E. R. (2008). Effect of crop rotation on specialization and genetic diversity of Macrophomina phaseolina . Tropical Plant Pathology , 33: 257-264. https://doi.org/10.1590/S1982-56762008000400001 Ambrósio, M. M. Q., Dantas, A. C. A., Martinez‐Perez, E., Medeiros, A. C., Nunes, G. H. S., & Picó, M. B. (2015). Screening a variable germplasm collection of Cucumis melo L. for seedling resistance to Macrophomina phaseolina . Euphytica , 206: 287-300. https://doi.org/10.1007/s10681-015-1452-x Barnett, H. L., & Hunter, B. B. (1998). Illustrated genera of imperfect fungi . 4th Ed. St. Paul, Minneapolis: American Phytopathological Society Press, 218. Bindal, S., Sheu, Z-M., Kenyon, L., Taher, D., & Rakha, M. (2023). Novel sources of resistance to fusarium wilt in Luffa species. Frontiers Plant Science , 14: 1-7. https://doi.org/10.3389/fpls.2023.1116006 Brito, A. C. Q., Mello, J. F., Michereff, S. J., Souza-Motta, C. M., & Machado, A. R. (2019). First report of Macrophomina pseudophaseolina causing stem dry rot in cassava in Brazil. Journal of Plant Pathology , 101: 1245. https://doi.org/10.1007/s42161-019-00309-3 Bruton, B. D., Popham, T. W., Garcia-Jiménez, J., Armengol, J., & Miller, M. E. (2000). Disease reaction among selected Cucurbitaceae to an Acremonium cucurbitacearum isolate from Texas. Horticultural Science , 35: 677-680. https://doi.org/10.21273/HORTSCI.35.4.677 Castellani, A. (1939). Viability of some pathogenic fungi in distilled water. Journal of Tropical Medicine & Hygiene , 24: 270-276. Cohen, R., Elkabetz, M., Paris, H. S., Gur, A., Dai, N., Rabinovitz, O., & Freeman, S. (2022). Occurrence of Macrophomina phaseolina in Israel: Challenges for disease management and crop germplasm enhancement. Plant Disease , 27: 50-74. https://doi.org/10.1094/PDIS-07-21-1390-FE Davis, A. R., Perkins-Veazie, P., Sakata, Y., López-Galarza, S., Maroto, J. V., Lee, S. G., Huh, Y. C., Sun, Z., Miguel, A., King, S. R., Cohen, R., & Lee, J. M. (2008). Cucurbit Grafting, Taylor & Francis Group. Critical Reviews in Plant Sciences , 27: 50-74. Guo Z., Qin, Y., LV, J., Wang, X., Dong, H., Dong, X., Zhang, T., Du, N., & Piao, F. (2023). Luffa rootstock enhances salt tolerance and improves yield and quality of grafted cucumber plants by reducing sodium transport to the shoot. Environmental Pollution , 316: 120-521. https://doi.org/10.1016/j.envpol.2022.120521 Gupta G. K., Sharma S. K., & Ramteke R. (2012). Biology, epidemiology and management of the pathogenic fungus Macrophomina phaseolina (Tassi) goid with special reference to charcoal rot of soybean ( Glycine max (L.) Merrill). Journal of Phytopatholgy , 160: 167-180. https://doi.org/10.1111/j.1439-0434.2012.01884.x Hilal, A. A., Abdel-Kader, D. A., Abo-El-Ela, A. M., & Nada, M. G. A. (2000). Soilborne fungal diseases of loofa ( Luffa aegyptiaca L.): new diseases in Egypt. Egyptian Journal of Agricultural Research , 78: 1823-1839. Islam, M. S., Haque, M. S., Islam, M. M., Emdad, E. M., Halim, A., Hossen, Q. M. M., Hossain, M. Z., Ahmed, B., Rahim, S., Rahman, M. S., Alam, M. M., Hou, S., Wan, X., Saito, J. A., & Alam, M. (2012). Tools to kill: Genome of one of the most destructive plant pathogenic fungi Macrophomina phaseolina . BMC Genomics , 13: 493. https://doi.org/10.1186/1471-2164-13-493 Jang, Y., Huh, Y., Park, D., Mun, B., Lee, S., & Um, Y. (2014). Greenhouse evaluation of melon rootstock resistance to Monosporascus root rot and vine decline as well as of yield and fruit quality in grafted 'inodorus' melons. Korean Journal of Horticultural Science & Technology , 32: 614-622. https://doi.org/10.7235/hort.2014.14065 Katuuramu D. N., Wechter, W. P., Washington, M. L., Horry, M., Cutulle, M. A., Jarret, R. L., & Levi, A. (2020). Phenotypic diversity for root traits and identification of superior germplasm for root breeding in watermelon. Horticultural Science , 55: 1272-1279. https://doi.org/10.21273/HORTSCI15093-20 Kwon, M-K., Hong, J-R., Kim, Y-H., & Kim, K-C. (2001). Soil-Environmental factors involved in the development of root rot/vine on Cucurbits caused by Monosporascus cannonballus. The Plant Pathology Journal , 17: 45-51. Li, H., Ahammed, G. J., Zhou, G., Xia, X., Zhou, J., Shi, K., Yu, J., & Zhou, Y. (2016). Unraveling main limiting sites of photosysnthesis under below-and above-ground heat stress in Cucumber and the alleviatory role of Luffa rootstock. Frontiers Plant Science , 7: 746. https://doi.org/10.3389/fpls.2016.00746 Machado, A. R., Pinho, D. B., Soares, D. J., Gomes, A. A. M., & Pereira, O. L. (2018). Bayesian analyses of five gene regions reveal a new phylogenetic species of Macrophomina associated with charcoal rot on oilseed crops in Brazil. European Journal of Plant Pathology , 153: 89-100. https://doi.org/10.1007/s10658-018-1545-1 Marquez, N., Giachero, M. L., Declerck, S., & Ducasse, D. A. (2021). Macrophomina phaseolina : General characteristics of pathogenicity and methods of control. Frontiers in Plant Science , 12: 634397. https://doi.org/10.3389/fpls.2021.634397 Medeiros, M. G., Neto, J. S. S. S., Oliveira, G. B. S., Torres, S. B., & Silveira, L. M. (2019). Physiological maturity of Luffa cylindrica (L.) Roem. seeds. Revista Ciência Agronômica , 50: 76-82. https://doi.org/10.5935/1806-6690.20190009 Mello, S. C. M., Reis, A., & Silva, J. B. T. (2011). Manual de curadores de germoplasma – Micro-organismos: fungos filamentosos . Embrapa Recursos Genéticos e Biotecnologia, 335. Embrapa Hortaliças, 134, 25 p. Namisy, A., Huang, J. H., Rakha, M., Hong, C. F., & Chung, W. H. (2023). Resistance to Fusarium oxysporum f. sp. luffae in Luffa germplasm despite hypocotyl colonization. Plant Disease , s/v.: 1-9. https://doi.org/10.1094/PDIS-08-22-1986-RE Negreiros, A. M. P., Sales Júnior, R., León, M., Melo, N. J. A., Michereff, S. J., Ambrósio, M. M. Q., Medeiros, H. L. S., & Armengol, J. (2019). Identification and pathogenicity of Macrophomina species collected from weeds in melon fields in Northeastern Brazil. Journal of Phytopathology , 137: 326-337. https://doi.org/10.1111/jph.12801 Negreiros, A. M. P., Melo, N. J. A., Ambrósio, M. M. Q., Nunes, G. H. S., & Sales Júnior, R. (2022). Growth rate, pathogenicity and fungicide sensitivity of Macrophomina spp. from weeds, melon and watermelon roots. Revista Caatinga , 35: 537-547. https://doi.org/10.1590/1983-21252022v35n304rc Nordey, T., Schwarz, D., Kenyon, L., Manickam, R., & Huat, J. (2020). Tapping the potential of grafting to improve the performance of vegetable cropping systems in sub-Saharan Africa. A review. Agronomy for Sustainable Development , 40: 1-18. https://doi.org/10.1007/s13593-020-00628-1 Oliveira, F. S., Nunes, M. C. C., Costa, F. M. C. D., Silva Neto, J. S. S., Silveira, L. M., & Torres, S. B. (2012). Superação de dormência em diferentes acessos de bucha. Horticultura Brasileira , 30: 2656-2662. (Supplement CD-ROM). Pereira, R. B., Pinheiro, J. B., & Carvalho, A. D. F. (2012). Identificação e manejo das principais doenças fúngicas do meloeiro . Brasília, DF: Embrapa (Circular Técnica, 112), s/v.: 5-6. Ravf, B. A., & Ahmad, I. (1998). Studies on correlation of seed infection to field incidence of Alternaria alternata and Macrophomina phaseolina in Sunflower. 13 th Iranian Plant Protection Congress-Karaj , 23-27. Salari, M., Panjehkeh, N., Nasirpoor, Z., & Abkhoo, J. (2012). Reaction of melon ( Cucumis melo L.) cultivars to soil-borne plant pathogenic fungi in Iran. African Journal of Biotechnology , 11: 15324-15329. Sales Júnior, R., Senhor, R. F., Michereff, S. J., & Negreiros, A. M. P. (2019). Reaction of genotypes to the root’s rot caused by Monosporascus. Revista Caatinga, 32: 288-294. https://doi.org/10.1590/1983-21252019v32n130rc Santos, K. M., Lima, G. S., Barros, A. P. O., Machado, A. R., Souza-Motta, C. M., Correia, C. C., & Michereff, S. J. (2020). Novel specific primers for rapid identification of Macrophomina species. European Journal of Plant Pathology , 156: 1213-1218. https://doi.org/10.1007/s10658-020-01952-8 Sarr, M. P., Ndiaye, M., Groenewald, J. Z., & Crous, P. W. (2014). Genetic diversity in Macrophomina phaseolina , the causal agent of charcoal rot. Pytopathologia Mediterranea , 53: 250-268. Seifert, K., Morgan-Jones, G., Gams, W., & Kendrick, B. (2011). The Genera of Hyphomycetes. CBS Biodiversity Series . CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands, 1-997. https://doi.org/10.3767/003158511X617435 Shahiba, A. M., & Arun, C. (2019). Grafting as a tool to improve drought tolerance in plants. In: Kumar, S. Ramesh et al . (ed.). Readers Shelf . J. V. Publishing House, 16: 1-68. Silva, F. A. Z., & Azevedo, C. A. V. (2016). The Assistat Software Version 7.7 and its use in the analysis of experimental data. African Journal Agricultural Research , 11: 3733-3740. Wu, H., Zhao, G., Gong, H., Li, J., Luo, C., He, X., Luo, S., Zheng, X., Liu, X., Guo, J., Chen, J., & Luo J. (2020). A high-quality sponge gourd ( Luffa cylindrica ) genome. Horticulture Research , 7: 128. https://doi.org/10.1038/s41438-020-00350-9 Additional Declarations No competing interests reported. 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NC: negative control; I: inoculated plant.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4094029/v1/8d200986c4b975bc3bf4a15a.png"},{"id":53790263,"identity":"ebd52921-54d1-40b9-bcc2-7f9d4a11f022","added_by":"auto","created_at":"2024-03-30 17:07:37","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2391734,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4094029/v1/f4dc8951-c34c-41a6-909d-3f899928a891.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Reaction of Luffa spp. to Macrophomina pseudophaseolina inoculation","fulltext":[{"header":"1 ǀ INTRODUCTION","content":"\u003cp\u003e \u003cem\u003eMacrophomina\u003c/em\u003e is a fungal genus belonging to the family Botryosphaeriaceae, class Dothideomycetes. The species \u003cem\u003eMacrophomina pseudophaseolina\u003c/em\u003e Crous, Sarr \u0026amp; Ndiaye, was first reported in Brazil in 2018, causing charcoal rot on oilseeds and jatropha seeds. More recently, it has been found on asymptomatic roots of weeds present in melon and watermelon production fields in the Northeast region of Brazil, as well as on cassava branches in Minas Gerais (Machado et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Brito et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Negreiros et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e (Tassi) G. Goid. and \u003cem\u003eM. pseudophaseolina\u003c/em\u003e are reported to be the most prevalent species in Brazil, both of which were considered pathogenic to melon trees in pathogenicity tests (Negreiros et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Negreiros et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe symptoms of \u003cem\u003eMacrophomina\u003c/em\u003e spp. generally include watery, light brown lesions with droplets of translucent exudate, which later darken, dry out, and become whitish, with longitudinal cracks. In severe cases, premature plant death occurs due to the blockage of the vascular system (xylem) caused by hyphal colonization, preventing the passage of water and nutrients to the plant (Islam et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Pereira et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Marquez et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). This genus exhibits good adaptation to regions with arid and semi-arid climates, thriving at temperatures between 26 and 38\u0026deg;C. It spreads through microsclerotia in the host's tissues or cultural remains as pycnidia, which can be observed as black dots on infected tissues (Almeida et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Gupta et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Pereira et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Sarr et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Negreiros et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCurrently, there are no registrations of synthetic active ingredients (fungicides) for the control of these phytopathogens in melon and watermelon production areas in Brazil. However, biological products based on \u003cem\u003eTrichoderma spp.\u003c/em\u003e and \u003cem\u003eBacillus spp.\u003c/em\u003e are registered for the control of \u003cem\u003eM. phaseolina\u003c/em\u003e in other crops (Agrofit, 2024). Nevertheless, the risks to the soil's micro and macrobiota from using these products are being investigated. Given this scenario, the most recommended approach involves managing cultural remains, using treated seeds, employing resistant genotypes, and utilizing rootstocks (Cohen et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Pereira et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Sales J\u0026uacute;nior et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe use of resistant rootstock is an effective strategy that enables plants to complete their life cycle in the field and is considered a less invasive solution for dynamic soil ecosystems (Jang et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Cohen et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, for the grafting technique to be effective, it is necessary to select plant material that is compatible with the target crop, easy to replicate, and provides resistance to these pathogens (Davis et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). According to Li et al. (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) and Bindal et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), \u003cem\u003eLuffa\u003c/em\u003e spp. seedlings can serve as rootstocks for other cucurbits such as pumpkin, cucumber, and melon. Hilal et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2000\u003c/span\u003e) tested fungi isolated from the roots and seeds of \u003cem\u003eLuffa aegyptiaca\u003c/em\u003e Mill and found that \u003cem\u003eM. phaseolina\u003c/em\u003e was not pathogenic to this crop. Subsequent studies also highlighted the resistance of this cucurbitaceous plant when inoculated with other root pathogens (Bruton et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Kwon et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Namisy et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Bindal et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe lack of knowledge about the influence of diseases transmitted by root pathogens has hindered the exploitation of this material for possible agricultural purposes (Bindal et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Further research is therefore needed to better understand these effects and develop appropriate management strategies. In this context, it is important to emphasize that the \u003cem\u003eLuffa\u003c/em\u003e genus shows variability in various characteristics and has a considerable number of resistance genes (Wu et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). It should also be noted that some species of this genus, such as \u003cem\u003eLuffa cylindrica\u003c/em\u003e M. Roem, are considered allogamous, reproducing preferentially by crossing. Therefore, evaluations are necessary to determine the resistance of these species to be used as rootstocks resistant to soil pathogens.\u003c/p\u003e \u003cp\u003eGiven the above, this study aimed to assess the pathogenicity of \u003cem\u003eM. pseudophaseolina\u003c/em\u003e on \u003cem\u003eLuffa\u003c/em\u003e spp. accessions to identify resistant materials that can later be used in breeding programs and/or directly as rootstocks for other cucurbits.\u003c/p\u003e"},{"header":"2 ǀ MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Obtaining and characterizing fungi from the roots of \u003cem\u003eLuffa\u003c/em\u003e spp.\u003c/h2\u003e \u003cp\u003eA prospecting study of fungi associated with roots of \u003cem\u003eLuffa\u003c/em\u003e spp. was conducted between February and May 2022 in the states of Cear\u0026aacute; (CE) and Rio Grande do Norte (RN). Nineteen root samples were collected from asymptomatic plants. Plant roots were washed in running water to remove soil residues, followed by immersion in a 1% sodium hypochlorite (NaClO) solution for one minute and rinsing with sterilized water to remove excess chlorine. Seven root fragments were placed in Petri dishes containing potato-dextrose-agar (PDA) medium with 0.5 g.L\u003csup\u003e-1\u003c/sup\u003e of tetracycline added (Mello et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe plates containing root fragments were incubated in a Biochemical Oxygen Demand (BOD) incubator at 28\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C in the dark for up to seven days. Fungal colonies growing on root fragments were transferred to Petri dishes containing PDA medium. Fungal isolates were identified to the genus level by preparing slides containing the fungal structures stained with lactophenol cotton blue for observation under an optical microscope and compared with descriptions in fungal identification keys (Barnett \u0026amp; Hunter, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Seifert et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe monohyphal purification technique was used to obtain pure cultures, which were then preserved using the Castellani method (1939). The isolates obtained were deposited in the fungal culture collection of the Phytopathology Laboratory II at Universidade Federal Rural do Semi-\u0026Aacute;rido (UFERSA).\u003c/p\u003e \u003cp\u003eFor the molecular characterization of \u003cem\u003eMacrophomina\u003c/em\u003e species, the polymerase chain reaction (PCR) technique with specific primers was used. Fungal isolates were cultivated in PDA medium with cellophane overlaid to allow the mycelia to be obtained, and subsequently the deoxyribonucleic acid (DNA) of each isolate to be extracted. Genomic DNA was extracted using the Wizard\u003csup\u003e\u0026reg;\u003c/sup\u003e genomic DNA purification kit (Promega Corporation, Madison, USA), following the manufacturer's protocol.\u003c/p\u003e \u003cp\u003ePCR amplification was carried out using 6.25 \u0026micro;L of GoTaq\u003csup\u003e\u0026reg;\u003c/sup\u003e Green Master Mix (2X) from Promega Corporation (Madison, USA), 4.25 \u0026micro;L of Milli-Q\u003csup\u003e\u0026reg;\u003c/sup\u003e water, 0.5 \u0026micro;l of each primer (forward and reverse), and 1 \u0026micro;l of DNA (25 ng/\u0026micro;L). Water was used for the negative control instead of DNA, while isolates previously identified with specific primers were used for the positive control. Specific primers from the translation elongation factor-1alpha (TEF1-α) locus were used for \u003cem\u003eM. phaseolina\u003c/em\u003e (MpTefF-AAACACACTTTTCGCACTCCTGC, MpTefR-TATGCTCGCAGAGAAGAACACGA), \u003cem\u003eM. pseudophaseolina\u003c/em\u003e (MsTefF-GCACACTTTTCGCGCTTCTGTA, MsTefR-TGTGCTCGCTGGGAAGAACATGA), and \u003cem\u003eM. euphorbiicola\u003c/em\u003e (MeTefF-AAGCATACTTTTCGTGCTCCTGC, MeTefR-AAAGGAACATGAGTGGCCAAAAA) (Santos et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThermal cycler was used with the following program: initial denaturation at 94\u0026deg;C for two minutes, 30 cycles of denaturation at 94\u0026deg;C for one minute, annealing at 63\u0026deg;C for 30 seconds, extension at 72\u0026deg;C for one minute, and a final extension at 72\u0026deg;C for ten minutes. After completing the amplification cycles, PCR products were separated by gel electrophoresis, loading 5 \u0026micro;L of the amplified DNA together with 3 \u0026micro;L of sample buffer\u0026thinsp;+\u0026thinsp;Gel Red 2X (Santos et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Finally, results were visualized under ultraviolet (UV) light. The species of the pathogen was confirmed by amplifying the products using primers specific to target species. All molecular techniques used to identify isolates were conducted in the Laboratory of Molecular Biology at the Universidade Federal do Cariri (UFCA), in the state of CE, Brazil.\u003c/p\u003e \u003cp\u003eFor the subsequent virulence test, a \u003cem\u003eMacrophomina\u003c/em\u003e isolate was selected, identified as \u003cem\u003eM. pseudophaseolina\u003c/em\u003e (RO-II), based on preliminary pathogenicity evaluations showing greater aggressiveness on \u003cem\u003eL. cylindrica\u003c/em\u003e and 'Natal' melon plants (unpublished data).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 \u003cem\u003eLuffa\u003c/em\u003e spp. accessions used to evaluate resistance to \u003cem\u003eM. pseudophaseolina\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eTwelve \u003cem\u003eLuffa\u003c/em\u003e spp. accessions from the Cucurbitaceae Germplasm Collection of the Department of Agronomic and Forestry Sciences of the Centre for Agrarian Sciences at UFERSA were used (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAccessions of \u003cem\u003eLuffa\u003c/em\u003e spp. used in the experiment\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAccession code\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIdentification\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eYear of collection\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePlace of collection\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB07\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eApodi/RN\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB24\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePetrolina/PE\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB26\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eApodi/RN\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB36\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSerra Talhada/PE\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB39\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2012\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePetrolina/PE\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB40\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2012\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eGov. Dix-Sept Rosado/RN\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB41\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2012\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eS\u0026atilde;o Jos\u0026eacute; do Belmonte/PE\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB44\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2012\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eApodi/RN\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB50\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eOroc\u0026oacute;/PE\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB51\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMonsenhor Tabosa/CE\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB52\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSerra Talhada/PE\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eAB55\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLuffa cylindrica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSerra Talhada/PE\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003csup\u003ea\u003c/sup\u003e Code of accessions in the Cucurbitaceae Germplasm Collection. \u003csup\u003eb\u003c/sup\u003e RN\u0026thinsp;=\u0026thinsp;Rio Grande do Norte State; PE\u0026thinsp;=\u0026thinsp;Pernambuco State; CE\u0026thinsp;=\u0026thinsp;Cear\u0026aacute; State.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Setting up and conducting the experiment\u003c/h2\u003e \u003cp\u003eTo assess the reaction of \u003cem\u003eLuffa\u003c/em\u003e species to RO-II, a test was conducted in a greenhouse at UFERSA's West Campus in the municipality of Mossor\u0026oacute; (RN), following a completely randomized design (CRD). Twelve accessions of \u003cem\u003eLuffa\u003c/em\u003e spp., yellow melon cultivar 'Natal', and watermelon cultivar 'Crimson Sweet' were inoculated with the fungal isolate, along with negative controls for each accession/cultivar. The experiment was repeated.\u003c/p\u003e \u003cp\u003e \u003cem\u003eLuffa\u003c/em\u003e spp. seeds underwent disinfection and dormancy overcoming processes (Oliveira et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Medeiros et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Seedlings were sown in polythene trays containing commercial substrate Tropstrato HT Hortali\u0026ccedil;as\u003csup\u003e\u0026reg;\u003c/sup\u003e (Vida Verde, Brazil) and watered daily. After showing the first definitive leaf, seedlings were transferred to 1 L pots containing a mixture of sand and autoclaved commercial substrate in a 3:1 ratio (v/v). Irrigation was carried out daily until the soil reached field capacity.\u003c/p\u003e \u003cp\u003eTen days after sowing, the \u003cem\u003eM. pseudophaseolina\u003c/em\u003e isolate was inoculated using the method described by Ambr\u0026oacute;sio et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), which involved inserting toothpicks colonized with the isolate's mycelium. Uninfested and autoclaved toothpicks were used as negative controls.\u003c/p\u003e \u003cp\u003eThe trial was evaluated 60 days after seedling transplantation. Plants were gently removed from pots, and root systems were carefully washed with running water. The variables analyzed included disease incidence and severity, average plant reaction to the pathogen, shoot and root lengths (SL and RL), fresh shoot and fresh root weights (FSW and FRW), and dry shoot and root weights (DSW and DRW).\u003c/p\u003e \u003cp\u003eDisease incidence was determined as the number of infected plants per treatment and expressed as percentage (%), while its severity was assessed by a diagrammatic grading scale. For \u003cem\u003eM. pseudophaseolina\u003c/em\u003e-inoculated plants, we used a modified scale described by Ravf \u0026amp; Ahmad (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1998\u003c/span\u003e), with scores ranging from 0 to 5, wherein: 0\u0026thinsp;=\u0026thinsp;no symptoms; 1\u0026thinsp;=\u0026thinsp;less than 3% of infected tissues; 2\u0026thinsp;=\u0026thinsp;3\u0026ndash;10% of infected tissues; 3\u0026thinsp;=\u0026thinsp;11\u0026ndash;25% of infected tissues; 4\u0026thinsp;=\u0026thinsp;25\u0026ndash;50% of infected tissues; and 5\u0026thinsp;=\u0026thinsp;more than 50% of infected tissues.\u003c/p\u003e \u003cp\u003eTo assess the overall reaction of plants to disease, we employed a scale outlined by Salari et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). This scale calculates the average score per treatment by summing the scores assigned to each individual plant, dividing by the total number of plants evaluated, and subsequently categorizing the results as follows: 0\u0026thinsp;=\u0026thinsp;immune (I); 0.1-1.0\u0026thinsp;=\u0026thinsp;highly resistant (HR); 1.1-2.0\u0026thinsp;=\u0026thinsp;moderately resistant (MR); 2.1-4.0\u0026thinsp;=\u0026thinsp;susceptible (SU); and 4.1-5.0\u0026thinsp;=\u0026thinsp;highly susceptible (HS).\u003c/p\u003e \u003cp\u003eShoot and root length were measured using a tape measure (cm). Fresh and dry weights of shoots and roots were determined using an analytical balance (g). To obtain the dry weights, each plant was placed individually in paper bags and subjected to a forced air circulation oven at 70\u0026deg;C until a consistent dry weight was achieved.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Statistical analysis\u003c/h2\u003e \u003cp\u003eIncidence and severity results were analyzed using the non-parametric Kruskal-Wallis test at a 5% probability level. Length, fresh and dry weights of shoots and roots were analyzed using ANOVA, and means were compared using the Scott-Knott test at a 5% probability level. Statistical analyses were conducted using Assistat 7.7 software (Silva \u0026amp; Azevedo, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e"},{"header":"3 ǀ RESULTS AND DISCUSSION","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Isolation and characterization of root-associated fungi in \u003cem\u003eLuffa\u003c/em\u003e spp.\u003c/h2\u003e \u003cp\u003eFrom asymptomatic \u003cem\u003eLuffa\u003c/em\u003e spp. roots, nineteen fungal isolates were obtained. Using an identification key, we successfully classified eight of these isolates to the genus level, namely: \u003cem\u003eLasiodiplodia\u003c/em\u003e sp. (5.0%), \u003cem\u003eChaetomium\u003c/em\u003e sp. (5.0%), \u003cem\u003eRhizoctonia\u003c/em\u003e sp. (16.0%), and \u003cem\u003eMacrophomina\u003c/em\u003e sp. (16.0%) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). However, conventional morphological identification methods proved inadequate for the remaining eleven isolates.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFurther characterization of the three \u003cem\u003eMacrophomina\u003c/em\u003e isolates identified through morphological features confirmed them as \u003cem\u003eM. pseudophaseolina\u003c/em\u003e using PCR analysis (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eFungal genera collected from \u003cem\u003eLuffa\u003c/em\u003e spp. roots in the states of Cear\u0026aacute; (CE) and Rio Grande do Norte (RN)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIdentification code\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIsolates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLocation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eGeographical coordinates\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRO \u0026ndash; II\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMacrophomina pseudophaseolina\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBrazil, Rio Grande do Norte, Mossor\u0026oacute;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5\u0026deg;12'23.26\"S/ 37\u0026deg;19'45.70\"W\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRO \u0026ndash; III\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMacrophomina pseudophaseolina\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBrazil, Rio Grande do Norte, Mossor\u0026oacute;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5\u0026deg;12'24.91\"S/ 37\u0026deg;19'46.29\"W\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eQZ \u0026ndash; I\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMacrophomina pseudophaseolina\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBrazil, Rio Grande do Norte, Mossor\u0026oacute;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5\u0026deg;12'30.05\"S/ 37\u0026deg;20'11.99\"W\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOV \u0026ndash; I\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eRhizoctonia\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBrazil, Rio Grande do Norte, Mossor\u0026oacute;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5\u0026deg;12'25.92\"S/ 37\u0026deg;18'59.85\"W\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eRhizoctonia\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBrazil, Rio Grande do Norte, Mossor\u0026oacute;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5\u0026deg;12'20.23\"S/ 37\u0026deg;19'07.46\"W\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBR- I\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eRhizoctonia\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBrazil, Rio Grande do Norte, Mossor\u0026oacute;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5\u0026deg;13'39.38\"S/ 37\u0026deg;19'17.17\"W\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCA \u0026ndash; II\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLasiodiplodia\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBrazil, Rio Grande do Norte, Mossor\u0026oacute;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5\u0026deg;11'48.95\"S/ 37e19'14.38\"W\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eR5 \u0026ndash; II\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eChaetomium\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBrazil, Cear\u0026aacute;, Caucaia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3\u0026deg;43'55.13\"S/ 38\u0026deg;38'41.69\"W\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003csup\u003ea\u003c/sup\u003e Isolates deposited in the collection of fungal isolates at the Phytopathology Laboratory II of the Universidade Federal Rural do Semi-\u0026Aacute;rido (UFERSA).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Response of \u003cem\u003eLuffa\u003c/em\u003e spp. accessions to \u003cem\u003eM. pseudophaseolina\u003c/em\u003e inoculation\u003c/h2\u003e \u003cp\u003eData from both experiments were combined since no significant effect of the experiment repetitions was observed (ANOVA \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05) for all the variables analyzed. Inoculation of cucurbits by \u003cem\u003eM. pseudophaseolina\u003c/em\u003e revealed statistically significant differences for disease incidence and severity, as demonstrated by the Kruskal-Wallis test at a 5% probability (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.05) (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e), and for SL, RL, FSW, FRW, DSW, and DRW, as indicated by the Scott-Knott test at a 5% probability (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.05) (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDisease incidence and severity, and reaction class against \u003cem\u003eMacrophomina pseudophaseolina\u003c/em\u003e inoculation in accessions of \u003cem\u003eLuffa\u003c/em\u003e spp., \u003cem\u003eCucumis melo\u003c/em\u003e, and \u003cem\u003eCitrullus lanatus\u003c/em\u003e.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTreatment/ accessions\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eDisease incidence\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eDisease severity\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eReaction\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRank\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMean (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRank\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMean\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e45.1 a-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSU\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e46.5 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e63.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e37.1 a-c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e60.2 a-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSU\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e60.2 a-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSU\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.5 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11.5 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e85.4 cd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e93.0 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e5.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e85.0 cd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e4.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e93.0 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e5.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e39.5 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e32.5 a-c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e65.0 a-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSU\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e53.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e75.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e27.2 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMelon\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e75.4 b-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e4.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHS\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWatermelon\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e32.5 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e38.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21.1 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHR\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eχ \u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e65.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e80.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eχ 2\u0026thinsp;=\u0026thinsp;significant chi-squared values at 5% probability. \u003csup\u003ea\u003c/sup\u003e Values followed by the same letter within columns do not differ statistically from each other by the Kruskal-Wallis non-parametric test (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.05). \u003csup\u003eb\u003c/sup\u003e Average scores for all observations within each sample following the diagrammatic scale: 0 (no symptoms) and 5 (more than 50% of infected tissues) (Ravf \u0026amp; Ahmad, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1998\u003c/span\u003e). \u003csup\u003ec\u003c/sup\u003e Reaction to the disease according to Salari et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), wherein: I\u0026thinsp;=\u0026thinsp;immune; HR\u0026thinsp;=\u0026thinsp;highly resistant; MR\u0026thinsp;=\u0026thinsp;moderately resistant; SU\u0026thinsp;=\u0026thinsp;susceptible; and HS\u0026thinsp;=\u0026thinsp;highly susceptible. Data are average values from two experiments, each with four replicates (pots) per treatment and one plant per pot.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAverages of lengths and fresh and dry weights of shoots and roots in accessions of \u003cem\u003eLuffa\u003c/em\u003e spp., \u003cem\u003eCucumis melo\u003c/em\u003e, and \u003cem\u003eCitrullus lanatus\u003c/em\u003e inoculated with \u003cem\u003eMacrophomina pseudophaseolina\u003c/em\u003e.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment/accessions\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSL\u003csup\u003ea\u003c/sup\u003e (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRL\u003csup\u003eb\u003c/sup\u003e (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFSW\u003csup\u003ec\u003c/sup\u003e (g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eFRW\u003csup\u003ed\u003c/sup\u003e (g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eDSW\u003csup\u003ee\u003c/sup\u003e (g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eDRW\u003csup\u003ef\u003c/sup\u003e (g)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e111.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e25.5 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.0 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.7 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.8 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.4 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e139.5 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e29.4 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.7 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.2 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.6 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e112.9 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e25.7 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10.7 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.0 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.1 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.2 d\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e122.0 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e26.0 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e16.6 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.0 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.5 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e163.9 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e43.4 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e24.9 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.8 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.4 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.5 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e58.5 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18.1 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.2 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.0 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.2 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.2 d\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e47.0 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.3 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.5 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.5 f\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.3 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.1 e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e69.1 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.0 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.7 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.5 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.4 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.1 e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e63.0 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.5 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.2 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.3 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.2 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.2 d\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e156.5 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35.2 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21.4 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.2 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.0 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e115.2 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.0 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.1 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.2 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.5 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAB55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e113.2 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.0 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e16.0 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.7 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.0 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMelon\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e46.2 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e21.0 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.0 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.3 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.1 e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWatermelon\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e83.0 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e26.6 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e22.5 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.6 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.0 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.2 d\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCV (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e26.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e23.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e20.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e18.17\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eCV (%)\u0026thinsp;=\u0026thinsp;significant coefficients of variation; values followed by the same letter within columns do not differ statistically from each other by the Scott-Knott test (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.05). \u003csup\u003ea\u003c/sup\u003e Shoot length. \u003csup\u003eb\u003c/sup\u003e Root length. \u003csup\u003ec\u003c/sup\u003e Fresh shoot weight. \u003csup\u003ed\u003c/sup\u003e Fresh root weight. \u003csup\u003ee\u003c/sup\u003e Dry shoot weight. \u003csup\u003ef\u003c/sup\u003e Dry root weight.\u003c/p\u003e \u003cp\u003eFor disease incidence, treatments AB24 (63.0%), AB39 (0.0%), and AB51 (50.0%) showed no significant difference compared to Watermelon (38.0%). Notably, among the Melon treatments, only AB39 (0.0%) exhibited a statistically different result remained unaffected by the pathogen, indicating its immunity to the disease (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe reaction to \u003cem\u003eM. pseudophaseolina\u003c/em\u003e revealed that nine out of the 12 \u003cem\u003eLuffa\u003c/em\u003e spp. accessions exhibited a disease incidence of over 75% (AB07, AB26, AB36, AB40, AB41, AB44, AB50, AB52, and AB55) (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Remarkably, only accession AB39 remained unaffected by the disease. Comparatively, among the positive controls, Watermelon showed the least susceptibility. This aligns with findings by Negreiros et al. (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), who observed higher disease incidence on melon (40% and 80% for isolates CMM4771 and CMM4801, respectively) compared to watermelon (0% and 40%, respectively), supporting the results of this study.\u003c/p\u003e \u003cp\u003eRegarding disease severity, \u003cem\u003eLuffa\u003c/em\u003e spp. accessions AB40 (4.5), AB41 (5.0), AB44 (4.6), and AB50 (5.0) differed from Watermelon (0.5). However, when comparing treatments inoculated with \u003cem\u003eM. pseudophaseolina\u003c/em\u003e to Melon (4.1), only AB39 (0.0) showed a statistically significant difference (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eNotably, the infection by \u003cem\u003eM. pseudophaseolina\u003c/em\u003e in accessions AB41, AB44, and AB50 initiated around the 19th day after inoculation. Various structures such as microsclerotia and pycnidia were observed in close proximity to the plant's neck. Additionally, symptoms such as cracks in the hypocotyl and chlorosis on lower leaves appeared, ultimately resulting in the death of the plants (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eBased on the reaction classification proposed by Salari et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), AB39 demonstrated immunity to the pathogen, while AB55 exhibited a highly resistant reaction, akin to the positive control Watermelon. Treatments AB24 and AB51 were categorized as moderately resistant, whereas treatments AB07, AB26, AB36, and AB52 were deemed susceptible to the pathogen. Treatments AB40, AB41, AB44, AB50, and Melon were classified as highly susceptible (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eHilal et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2000\u003c/span\u003e) conducted inoculation trials on \u003cem\u003eL. aegyptiaca\u003c/em\u003e plants infected with \u003cem\u003eM. phaseolina\u003c/em\u003e and reported the absence of symptoms. However, when combined with another pathogen, \u003cem\u003ePythium\u003c/em\u003e sp., visible symptoms emerged. Interestingly, the authors noted that the same \u003cem\u003eM. phaseolina\u003c/em\u003e isolate (derived from \u003cem\u003eL. aegyptiaca\u003c/em\u003e) exhibited high pathogenicity when introduced to various other crops, including cotton 'Giza 75', cucumber 'Amira 2', okra 'Costa Dourada', peanut 'Giza 4', pumpkin 'Balady', sesame 'Giza 32', soya 'Crawford', zucchini 'Eskandarani', and watermelon 'Giza 1'.\u003c/p\u003e \u003cp\u003eWu et al. (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) noted that the significant genetic variability within \u003cem\u003eLuffa\u003c/em\u003e spp. may result in certain accessions possessing enriched resistance genes. These genes enhance the plants' capacity to withstand various environmental stresses, including water scarcity, temperature fluctuations, high humidity, and diseases.\u003c/p\u003e \u003cp\u003eNamisy et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) observed similar findings, noting variations in the reactions of different accessions when inoculated with \u003cem\u003eFusarium oxysporum\u003c/em\u003e f. sp. \u003cem\u003eluffae\u003c/em\u003e. The authors attributed these differences to the influence of plant genotype on disease susceptibility and resistance.\u003c/p\u003e \u003cp\u003eFor the biometric variable SL, treatments AB24, AB39, and AB51 demonstrated statistically superior performance compared to other treatments, with average values ranging from 139.5 to 163.9 cm, representing the highest recorded lengths. Conversely, treatments AB07, AB26, AB36, AB52, and AB55 significantly differed from both Melon and Watermelon, displaying lengths ranging from 111.5 to 122.0 cm. However, treatments AB40, AB41, AB44, and AB50 (ranging from 47.0 to 69.1 cm) did not exhibit significant differences compared to Melon and Watermelon (measuring 46.2 and 83.0 cm, respectively), indicating the shortest lengths.\u003c/p\u003e \u003cp\u003eAccession AB39 stood out statistically among all treatments for the variable RL (43.4 cm), FSW (24.9 g), FRW (9.8 g), and DRW (0.5 g), boasting the highest average values across these variables.\u003c/p\u003e \u003cp\u003eConversely, for the RL variable, treatments AB40, AB41, AB44, and Melon showed significantly lower averages (ranging from 16.0 to 21.0 cm), indicating a notable reduction in this characteristic. Similarly, in terms of FSW, treatments AB40, AB41, AB44, and AB50 exhibited inferior values compared to other treatments, with averages ranging from 4.2 to 6.7 g. Regarding FRW, AB41 displayed the lowest average value (0.5 g) among treatments, indicating a greater reduction in the root system. With respect to DRW, the lowest recorded weight was 0.1 g for treatments TB41, TB44, and Melon.\u003c/p\u003e \u003cp\u003eConcerning DSW, treatments AB07, AB24, AB26, AB36, AB39, AB51, AB52, and Watermelon demonstrated statistically superior performance, ranging from 2.1 to 3.4 g. Conversely, treatments AB40, AB41, AB44, AB50, and AB55 exhibited average values ranging from 1.2 to 2.0 g, which differed significantly from the Watermelon treatment but not from the Melon treatment.\u003c/p\u003e \u003cp\u003eAs \u003cem\u003eMacrophomina\u003c/em\u003e spp. is a fungus naturally present in the soil and associated with its chemical and physical conditions, besides constantly interacting with other pathosystems, any strategy for disease management should aim to minimize alterations or manipulations of environmental and natural resources (Marquez et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In this sense, pathogenicity studies play a crucial role in identifying resistant plant materials, as controlling \u003cem\u003eMacrophomina\u003c/em\u003e spp. is inherently complex. Therefore, it is vital to implement one or a combination of control methods to mitigate the detrimental effects of phytopathogen attacks on vulnerable crops, thereby reducing inoculum pressure and the incidence of disease (Agrofit, 2024).\u003c/p\u003e \u003cp\u003eGrafting stands out as one of the most effective and environmentally friendly methods for controlling \u003cem\u003eMacrophomina\u003c/em\u003e spp. in agricultural fields. Cohen et al. (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) emphasized its success in select regions of Israel, where it has effectively managed this pathogen in melon and watermelon crops by utilizing pumpkin rootstock resistant to the disease. However, this approach necessitates careful selection of plant materials that not only align with target crops but also offer replicability and resistance to pathogens (Davis et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). In recent years, researchers have explored \u003cem\u003eLuffa\u003c/em\u003e species as potential rootstocks for cucurbits such as pumpkin, cucumber, and melon (Li et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Shahiba \u0026amp; Chacko, 2019; Nordey et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Guo et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), making accession AB39 particularly promising for breeding programs aimed at developing resistant cultivars.\u003c/p\u003e \u003cp\u003eMoreover, it is noteworthy that this study marks the first documentation of \u003cem\u003eLuffa\u003c/em\u003e spp. acting as an alternative host for \u003cem\u003eM. pseudophaseolina\u003c/em\u003e, thus contributing significantly to our understanding of the interaction between \u003cem\u003eM. pseudophaseolina\u003c/em\u003e and \u003cem\u003eLuffa\u003c/em\u003e species. These findings offer valuable insights for managing the disease in other cucurbit crops. Additionally, accession AB39 exhibited the highest averages for RL and FRW among all treatments. As highlighted by Katuuramu et al. (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), robust root development enhances nutrient and water absorption while mitigating severity of diseases transmitted by soil pathogens.\u003c/p\u003e \u003c/div\u003e"},{"header":"4 ǀ CONCLUSION","content":"\u003cp\u003eAccession AB39 was classified as \"immune\" to the pathogen. This is the first report of \u003cem\u003eLuffa\u003c/em\u003e spp. acting as an alternative host for \u003cem\u003eMacrophomina pseudophaseolina\u003c/em\u003e.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was funded by the Conselho Nacional de Desenvolvimento Cient\u0026iacute;fico e Tecnol\u0026oacute;gico (CNPq) \u0026ndash; Brazil.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors of the manuscript declare that there are no personal, commercial, academic, political, or financial conflicts of interest in the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAMPN, LMS, and RSJ contributed to the conception and design of the study. DMV, AMPN, ALAC, and CPSSA set up the tests and collected data. DMV and AMPN performed the statistical analyses. DMV, AMPN, LMS, ALAC, and RSJ assisted with the writing, proofreading, and editing of the manuscript. All authors contributed to the data analysis, interpretation of the results, and writing of the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAGROFIT. \u003cem\u003eSistemas de agrot\u0026oacute;xicos fitossanit\u0026aacute;rios\u003c/em\u003e.\u0026nbsp;Available at: \u0026lt;http://extranet.agricultura.gov.br/agrofit_cons/principal_agrofit_cons\u0026gt;. Accessed on: Jan 23, 2024.\u003c/li\u003e\n \u003cli\u003eAlmeida, A. M. R., Sosa-Gomez, D. R., Binneck, E., Marin, S. R. R., Zucchi, M. I., Abdelnoor, R. V., \u0026amp; Souto, E. R. (2008). Effect of crop rotation on specialization and genetic diversity of \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e. \u003cem\u003eTropical Plant Pathology\u003c/em\u003e,\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e33: 257-264. https://doi.org/10.1590/S1982-56762008000400001\u003c/li\u003e\n \u003cli\u003eAmbr\u0026oacute;sio, M. M. Q., Dantas, A. C. A., Martinez‐Perez, E., Medeiros, A. C., Nunes, G. H. S., \u0026amp; Pic\u0026oacute;, M. B. (2015). Screening a variable germplasm collection of \u003cem\u003eCucumis melo\u003c/em\u003e L. for seedling resistance to \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e. \u003cem\u003eEuphytica\u003c/em\u003e, 206: 287-300. https://doi.org/10.1007/s10681-015-1452-x\u003c/li\u003e\n \u003cli\u003eBarnett, H. L., \u0026amp; Hunter, B. B. (1998). \u003cem\u003eIllustrated genera of imperfect\u0026nbsp;fungi\u003c/em\u003e. 4th Ed. St. Paul, Minneapolis: American Phytopathological Society Press, 218.\u003c/li\u003e\n \u003cli\u003eBindal, S., Sheu, Z-M., Kenyon, L., Taher, D., \u0026amp; Rakha, M. (2023). Novel sources of resistance to fusarium wilt in \u003cem\u003eLuffa\u003c/em\u003e species. \u003cem\u003eFrontiers Plant Science\u003c/em\u003e, 14: 1-7. https://doi.org/10.3389/fpls.2023.1116006\u003c/li\u003e\n \u003cli\u003eBrito, A. C. Q., Mello, J. F., Michereff, S. J., Souza-Motta, C. M., \u0026amp; Machado, A. R. (2019). First report of \u003cem\u003eMacrophomina pseudophaseolina\u003c/em\u003e causing stem dry rot in cassava in Brazil. \u003cem\u003eJournal of Plant Pathology\u003c/em\u003e, 101: 1245. https://doi.org/10.1007/s42161-019-00309-3\u003c/li\u003e\n \u003cli\u003eBruton, B. D., Popham, T. W., Garcia-Jim\u0026eacute;nez, J., Armengol, J., \u0026amp; Miller, M. E. (2000). Disease reaction among selected Cucurbitaceae to an \u003cem\u003eAcremonium cucurbitacearum\u003c/em\u003e isolate from Texas. \u003cem\u003eHorticultural Science\u003c/em\u003e, 35: 677-680. https://doi.org/10.21273/HORTSCI.35.4.677\u003c/li\u003e\n \u003cli\u003eCastellani, A. (1939). Viability of some pathogenic fungi in distilled water. \u003cem\u003eJournal of Tropical Medicine \u0026amp; Hygiene\u003c/em\u003e, 24: 270-276.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eCohen, R., Elkabetz, M., Paris, H. S., Gur, A., Dai, N., Rabinovitz, O., \u0026amp; Freeman, S. (2022). Occurrence of \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e in Israel: Challenges for disease management and crop germplasm enhancement. \u003cem\u003ePlant Disease\u003c/em\u003e,\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e27: 50-74. https://doi.org/10.1094/PDIS-07-21-1390-FE\u003c/li\u003e\n \u003cli\u003eDavis, A. R., Perkins-Veazie, P., Sakata, Y., L\u0026oacute;pez-Galarza, S., Maroto, J. V., Lee, S. G., Huh, Y. C., Sun, Z., Miguel, A., King, S. R., Cohen, R., \u0026amp; Lee, J. M. (2008). Cucurbit Grafting, Taylor \u0026amp; Francis Group. \u003cem\u003eCritical Reviews in Plant Sciences\u003c/em\u003e, 27: 50-74.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eGuo Z., Qin, Y., LV, J., Wang, X., Dong, H., Dong, X., Zhang, T., Du, N., \u0026amp; Piao, F. (2023). \u003cem\u003eLuffa\u003c/em\u003e rootstock enhances salt tolerance and improves yield and quality of grafted cucumber plants by reducing sodium transport to the shoot. \u003cem\u003eEnvironmental Pollution\u003c/em\u003e, 316: 120-521. https://doi.org/10.1016/j.envpol.2022.120521\u003c/li\u003e\n \u003cli\u003eGupta G. K., Sharma S. K., \u0026amp; Ramteke R. (2012). Biology, epidemiology and management of the pathogenic fungus \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e (Tassi) goid with special reference to charcoal rot of soybean (\u003cem\u003eGlycine max\u003c/em\u003e (L.) Merrill). \u003cem\u003eJournal of Phytopatholgy\u003c/em\u003e, 160: 167-180. https://doi.org/10.1111/j.1439-0434.2012.01884.x\u003c/li\u003e\n \u003cli\u003eHilal, A. A., Abdel-Kader, D. A., Abo-El-Ela, A. M., \u0026amp; Nada, M. G. A. (2000). Soilborne fungal diseases of loofa (\u003cem\u003eLuffa aegyptiaca\u003c/em\u003e L.): new diseases in Egypt. \u003cem\u003eEgyptian Journal of Agricultural Research\u003c/em\u003e, 78: 1823-1839.\u003c/li\u003e\n \u003cli\u003eIslam, M. S., Haque, M. S., Islam, M. M., Emdad, E. M., Halim, A., Hossen, Q. M. M., Hossain, M. Z., Ahmed, B., Rahim, S., Rahman, M. S., Alam, M. M., Hou, S., Wan, X., Saito, J. A., \u0026amp; Alam, M. (2012). Tools to kill: Genome of one of the most destructive plant pathogenic fungi \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e. \u003cem\u003eBMC Genomics\u003c/em\u003e, 13: 493. https://doi.org/10.1186/1471-2164-13-493\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eJang, Y., Huh, Y., Park, D., Mun, B., Lee, S., \u0026amp; Um, Y. (2014). Greenhouse evaluation of melon rootstock resistance to \u003cem\u003eMonosporascus\u003c/em\u003e root rot and vine decline as well as of yield and fruit quality in grafted \u0026apos;inodorus\u0026apos; melons. \u003cem\u003eKorean Journal of Horticultural Science \u0026amp; Technology\u003c/em\u003e, 32: 614-622. https://doi.org/10.7235/hort.2014.14065\u003c/li\u003e\n \u003cli\u003eKatuuramu D. N., Wechter, W. P., Washington, M. L., Horry, M., Cutulle, M. A., Jarret, R. L., \u0026amp; Levi, A. (2020). Phenotypic diversity for root traits and identification of superior germplasm for root breeding in watermelon. \u003cem\u003eHorticultural Science\u003c/em\u003e, 55: 1272-1279. https://doi.org/10.21273/HORTSCI15093-20\u003c/li\u003e\n \u003cli\u003eKwon, M-K., Hong, J-R., Kim, Y-H., \u0026amp; Kim, K-C. (2001). Soil-Environmental factors involved in the development of root rot/vine on Cucurbits caused by \u003cem\u003eMonosporascus cannonballus.\u0026nbsp;\u003c/em\u003e\u003cem\u003eThe Plant Pathology Journal\u003c/em\u003e, 17: 45-51.\u003c/li\u003e\n \u003cli\u003eLi, H., Ahammed, G. J., Zhou, G., Xia, X., Zhou, J., Shi, K., Yu, J., \u0026amp; Zhou, Y. (2016). Unraveling main limiting sites of photosysnthesis under below-and above-ground heat stress in Cucumber and the alleviatory role of \u003cem\u003eLuffa\u0026nbsp;\u003c/em\u003erootstock.\u0026nbsp;\u003cem\u003eFrontiers Plant Science\u003c/em\u003e, 7: 746. https://doi.org/10.3389/fpls.2016.00746\u003c/li\u003e\n \u003cli\u003eMachado, A. R., Pinho, D. B., Soares, D. J., Gomes, A. A. M., \u0026amp; Pereira, O. L. (2018).\u0026nbsp;Bayesian analyses of five gene regions reveal a new phylogenetic species of \u003cem\u003eMacrophomina\u003c/em\u003e associated with charcoal rot on oilseed crops in Brazil. \u003cem\u003eEuropean Journal of Plant Pathology\u003c/em\u003e, 153: 89-100. https://doi.org/10.1007/s10658-018-1545-1\u003c/li\u003e\n \u003cli\u003eMarquez, N., Giachero, M. L., Declerck, S., \u0026amp; Ducasse, D. A. (2021). \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e: General characteristics of pathogenicity and methods of control. \u003cem\u003eFrontiers in Plant Science\u003c/em\u003e, 12: 634397. https://doi.org/10.3389/fpls.2021.634397\u003c/li\u003e\n \u003cli\u003eMedeiros, M. G., Neto, J. S. S. S., Oliveira, G. B. S., Torres, S. B., \u0026amp; Silveira, L. M. (2019).\u0026nbsp;Physiological maturity of \u003cem\u003eLuffa cylindrica\u003c/em\u003e (L.) Roem. seeds. \u003cem\u003eRevista Ci\u0026ecirc;ncia Agron\u0026ocirc;mica\u003c/em\u003e, 50: 76-82. https://doi.org/10.5935/1806-6690.20190009\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eMello, S. C. M., Reis, A., \u0026amp; Silva, J. B. T. (2011). \u003cem\u003eManual de curadores de germoplasma \u0026ndash; Micro-organismos: fungos filamentosos\u003c/em\u003e. Embrapa Recursos Gen\u0026eacute;ticos e Biotecnologia, 335. Embrapa Hortali\u0026ccedil;as, 134, 25 p.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eNamisy, A., Huang, J. H., Rakha, M., Hong, C. F., \u0026amp; Chung, W. H. (2023).\u0026nbsp;Resistance to \u003cem\u003eFusarium oxysporum\u003c/em\u003e f. sp. \u003cem\u003eluffae\u003c/em\u003e in \u003cem\u003eLuffa\u003c/em\u003e germplasm despite hypocotyl colonization. \u003cem\u003ePlant Disease\u003c/em\u003e,\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003es/v.: 1-9. https://doi.org/10.1094/PDIS-08-22-1986-RE\u003c/li\u003e\n \u003cli\u003eNegreiros, A. M. P., Sales J\u0026uacute;nior, R., Le\u0026oacute;n, M., Melo, N. J. A., Michereff, S. J., Ambr\u0026oacute;sio, M. M. Q., Medeiros, H. L. S., \u0026amp; Armengol, J. (2019). Identification and pathogenicity of \u003cem\u003eMacrophomina\u003c/em\u003e species collected from weeds in melon fields in Northeastern Brazil. \u003cem\u003eJournal of Phytopathology\u003c/em\u003e, 137: 326-337. https://doi.org/10.1111/jph.12801\u003c/li\u003e\n \u003cli\u003eNegreiros, A. M. P., Melo, N. J. A., Ambr\u0026oacute;sio, M. M. Q., Nunes, G. H. S., \u0026amp; Sales J\u0026uacute;nior, R. (2022). Growth rate, pathogenicity and fungicide sensitivity of \u003cem\u003eMacrophomina\u0026nbsp;\u003c/em\u003espp. from weeds, melon and watermelon roots. \u003cem\u003eRevista Caatinga\u003c/em\u003e, 35: 537-547. https://doi.org/10.1590/1983-21252022v35n304rc\u003c/li\u003e\n \u003cli\u003eNordey, T., Schwarz, D., Kenyon, L., Manickam, R., \u0026amp; Huat, J. (2020). Tapping the potential of grafting to improve the performance of vegetable cropping systems in sub-Saharan Africa. A review. \u003cem\u003eAgronomy for Sustainable Development\u003c/em\u003e,\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e40: 1-18. https://doi.org/10.1007/s13593-020-00628-1\u003c/li\u003e\n \u003cli\u003eOliveira, F. S., Nunes, M. C. C., Costa, F. M. C. D., Silva Neto, J. S. S., Silveira, L. M., \u0026amp; Torres, S. B. (2012). Supera\u0026ccedil;\u0026atilde;o de dorm\u0026ecirc;ncia em diferentes acessos de bucha. \u003cem\u003eHorticultura Brasileira\u003c/em\u003e, 30: 2656-2662. (Supplement CD-ROM).\u003c/li\u003e\n \u003cli\u003ePereira, R. B., Pinheiro, J. B., \u0026amp; Carvalho, A. D. F. (2012). \u003cem\u003eIdentifica\u0026ccedil;\u0026atilde;o e manejo das principais doen\u0026ccedil;as f\u0026uacute;ngicas do meloeiro\u003c/em\u003e. Bras\u0026iacute;lia, DF: Embrapa (Circular T\u0026eacute;cnica, 112), s/v.:\u0026nbsp;5-6.\u003c/li\u003e\n \u003cli\u003eRavf, B. A., \u0026amp; Ahmad, I. (1998).\u0026nbsp;Studies on correlation of seed infection to field incidence of \u003cem\u003eAlternaria alternata\u003c/em\u003e and \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e in Sunflower. \u003cem\u003e13 th Iranian Plant Protection Congress-Karaj\u003c/em\u003e, 23-27.\u003c/li\u003e\n \u003cli\u003eSalari, M., Panjehkeh, N., Nasirpoor, Z., \u0026amp; Abkhoo, J. (2012). Reaction of melon (\u003cem\u003eCucumis melo\u003c/em\u003e L.) cultivars to soil-borne plant pathogenic fungi in Iran. \u003cem\u003eAfrican Journal of Biotechnology\u003c/em\u003e, 11: 15324-15329.\u003c/li\u003e\n \u003cli\u003eSales J\u0026uacute;nior, R., Senhor, R. F., Michereff, S. J., \u0026amp; Negreiros, A. M. P. (2019). Reaction of genotypes to the root\u0026rsquo;s rot caused by \u003cem\u003eMonosporascus.\u0026nbsp;\u003c/em\u003eRevista Caatinga, 32: 288-294. https://doi.org/10.1590/1983-21252019v32n130rc\u003c/li\u003e\n \u003cli\u003eSantos, K. M., Lima, G. S., Barros, A. P. O., Machado, A. R., Souza-Motta, C. M., Correia, C. C., \u0026amp; Michereff, S. J. (2020).\u0026nbsp;Novel specific primers for rapid identification of \u003cem\u003eMacrophomina\u003c/em\u003e species. \u003cem\u003eEuropean Journal of Plant Pathology\u003c/em\u003e, 156: 1213-1218. https://doi.org/10.1007/s10658-020-01952-8\u003c/li\u003e\n \u003cli\u003eSarr, M. P., Ndiaye, M., Groenewald, J. Z., \u0026amp; Crous, P. W. (2014). Genetic diversity in \u003cem\u003eMacrophomina phaseolina\u003c/em\u003e, the causal agent of charcoal rot. \u003cem\u003ePytopathologia Mediterranea\u003c/em\u003e,\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e53: 250-268.\u003c/li\u003e\n \u003cli\u003eSeifert, K., Morgan-Jones, G., Gams, W., \u0026amp; Kendrick, B. (2011). The Genera of Hyphomycetes. \u003cem\u003eCBS Biodiversity Series\u003c/em\u003e. CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands, 1-997. https://doi.org/10.3767/003158511X617435\u003c/li\u003e\n \u003cli\u003eShahiba, A. M., \u0026amp; Arun, C. (2019). Grafting as a tool to improve drought tolerance in plants. In: Kumar, S. Ramesh \u003cem\u003eet al\u003c/em\u003e. (ed.). \u003cem\u003eReaders Shelf\u003c/em\u003e. J. V. Publishing House, 16: 1-68.\u003c/li\u003e\n \u003cli\u003eSilva, F. A. Z., \u0026amp; Azevedo, C. A. V. (2016).\u0026nbsp;The Assistat Software Version 7.7 and its use in the analysis of experimental data. \u003cem\u003eAfrican Journal Agricultural Research\u003c/em\u003e, 11: 3733-3740.\u003c/li\u003e\n \u003cli\u003eWu, H., Zhao, G., Gong, H., Li, J., Luo, C., He, X., Luo, S., Zheng, X., Liu, X., Guo, J., Chen, J., \u0026amp; Luo J. (2020). A high-quality sponge gourd (\u003cem\u003eLuffa cylindrica\u003c/em\u003e) genome. \u003cem\u003eHorticulture Research\u003c/em\u003e,\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e7: 128. https://doi.org/10.1038/s41438-020-00350-9\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Loofah, Cucurbitaceae, Reaction of accessions, Root pathogens","lastPublishedDoi":"10.21203/rs.3.rs-4094029/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4094029/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe genus \u003cem\u003eMacrophomina\u003c/em\u003e is responsible for causing diseases in various species of the Cucurbitaceae family. This study aimed to evaluate the reaction of \u003cem\u003eLuffa\u003c/em\u003e species when associated with \u003cem\u003eM. pseudophaseolina\u003c/em\u003e. Initially, a prospection study was carried out to identify fungi associated with \u003cem\u003eLuffa\u003c/em\u003e roots. Three isolates belonging to the \u003cem\u003eM. pseudophaseolina\u003c/em\u003e species were identified and confirmed by DNA extraction and PCR amplification, using specific primers. One isolate was selected to test pathogenicity on \u003cem\u003eLuffa\u003c/em\u003e. Twelve \u003cem\u003eLuffa\u003c/em\u003e spp. accessions from the cucurbitaceous germplasm collection of Universidade Federal Rural do Semi-\u0026Aacute;rido were used, along with two positive controls: melon and watermelon plants. Inoculation was performed using the infested toothpick method. The plants were maintained under greenhouse conditions in a completely randomized design with four replications. The trial was repeated. The following variables were assessed: disease incidence and severity, reaction class, shoot length (SL), root length (RL), fresh shoot weight (FSW), dry shoot weight (DSW), fresh root weight (FRW), and dry root weight (DRW). Accession AB39 proved to be immune to the pathogen, as well as exhibiting the highest values of SL (163.9 cm), RL (43.4 cm), FSW (24.9 g), FRW (9.8 g), DSW (3.4 g), and DRW (0.5 g). These results indicate that the immune resistance of AB39 makes this accession promising, both in terms of disease resistance and vegetative growth, standing out as the best option among the accessions evaluated. Additionally, this is the first report of \u003cem\u003eLuffa\u003c/em\u003e spp. acting as an alternative host of \u003cem\u003eM. pseudophaseolina\u003c/em\u003e.\u003c/p\u003e","manuscriptTitle":"Reaction of Luffa spp. to Macrophomina pseudophaseolina inoculation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-19 09:49:20","doi":"10.21203/rs.3.rs-4094029/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":"f7562b33-808e-45e6-9c50-6a0551e4dd7a","owner":[],"postedDate":"March 19th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-03-30T16:59:30+00:00","versionOfRecord":[],"versionCreatedAt":"2024-03-19 09:49:20","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4094029","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4094029","identity":"rs-4094029","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}