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D’Hauwers, W.A.A. Tjalma, U. Punjabi and C.E. Depuydt",authors:[{id:"73860",title:"Mrs.",name:"Kathleen",middleName:null,surname:"DHauwers",fullName:"Kathleen DHauwers",slug:"kathleen-dhauwers"},{id:"78387",title:"Prof.",name:"Wiebren",middleName:null,surname:"Tjalma",fullName:"Wiebren Tjalma",slug:"wiebren-tjalma"},{id:"122938",title:"Dr.",name:"Usha",middleName:null,surname:"Punjabi",fullName:"Usha Punjabi",slug:"usha-punjabi"},{id:"122939",title:"Dr.",name:"Christophe",middleName:null,surname:"Depuydt",fullName:"Christophe Depuydt",slug:"christophe-depuydt"}]},{id:"26308",title:"The Impact of Human Papillomavirus on Cancer Risk in Penile Cancer",slug:"the-impact-of-human-papillomavirus-on-cancer-risk-in-penile-cancer",signatures:"Angela Adamski da Silva Reis and Aparecido Divino da Cruz",authors:[{id:"68829",title:"Dr.",name:"Angela",middleName:"Adamski Da Silva",surname:"Reis",fullName:"Angela Reis",slug:"angela-reis"},{id:"121027",title:"Prof.",name:"Aparecido Divino",middleName:null,surname:"Da Cruz",fullName:"Aparecido Divino Da Cruz",slug:"aparecido-divino-da-cruz"}]}]}]},onlineFirst:{chapter:{type:"chapter",id:"73149",title:"Assessment of Biocontrol Potential of Arbuscular Mycorrhizal (Glomus spp.) against Damping-off Disease (Rhizoctonia solani) on Cucumber",doi:"10.5772/intechopen.93313",slug:"assessment-of-biocontrol-potential-of-arbuscular-mycorrhizal-glomus-spp-against-damping-off-disease-",body:'\n
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1. Introduction
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\nRhizoctonia solani Kühn, the causative agent of damping-off disease in a variety of crop plants such as cucumber, is an economical important soil-borne pathogen [1, 2]. R. solani fungus is considered as a difficult pathogen to control due to several characters such as the great variability in the pathogen population, a wide host range, and long-term survival in soil [3]. Further, some cultural practices including the crop rotation, sanitation, and soil solarization with R. solani are not sufficiently effective because the pathogen is able to survive for many years in soil. The application of chemical pesticides, mainly methyl bromide, is the most reliable method to control R. solani; however, it causes serious risks including polluting the air, damaging the environment, building fungicides’ resistance of pathogen, and harming the human health [4, 5]. Therefore, the biological control method becomes an important component of the disease management to increase crop production and food safety [6].
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The biological control becomes an important target of many researchers in the field of biological and agricultural sciences [5]. Biocontrol agents use different mechanisms of action against fungal pathogens, such as antimicrobial compound production activity, mycoparasitism or hyperparasitism, cell wall-lytic enzyme activity, and the application of systemic resistance (ISR) activity [7]. In addition, some biocontrol agents are capable of improving some aspects of plant growth, such as the germination rate, shoot and root weight, nutrients’ uptake, and yield [8].
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Arbuscular mycorrhizal (AM) fungi have been known to form a symbiotic relationship with around 80% of vascular plants. The symbiotic relationship can provide the plant with many benefits, including enhancement of plant growth and germination rates, increasing supplement of water and nutrients [9, 10]. In return, the AM fungi are completely dependable on the nutrients that are coming from the living root system [9]. In addition, AM fungi have been known to increase the host’s resistance to a wide range of fungal and bacteria pathogens, especially rot pathogens [11]. The aim of this study was to examine the influence of different species of arbuscular mycorrhizal (AM) fungi (Glomus spp.) to promote systemic resistance against the disease agent of damping-off disease (R. solani Kühn) on cucumber (Cucumis sativus L.).
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2. Materials and methods
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Infected samples were brought from cucumber plants with wilting, yellowing, and dwarfing symptoms from a field related to the College of agriculture, University of Al-Qadisiyah. The plants were washed with sterilized water to remove soil residues and were cut to small pieces. Then, the samples were sterilized with sodium hypochlorite (NaCIO) 1% for 2 min, washed with sterilized water twice, and dried with filter papers. Nine petri dishes of potato dextrose agar (PDA) were inoculated with five pieces of the infected plants and incubated for 3 days at 25°C. Soil samples were diluted for pathogen isolation and the petri dishes were incubated at 27°C. Both plant and soil samples were kept in a refrigerator at 4°C and diagnosed using classification keys [12].
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Isolated pathogens were stored at 4°C prior to analysis and incubated at 25°C for 3 days. From the colony edge, four populated agar disks (7 mm) were cut and mixed in a 250 ml flask containing 100 ml of potato dextrose broth and 25 mg of chloramphenicol [13]. Sterilized soils were separated on each pot (3 kg) and inoculated with 1 ml from pathogen broth culture, and sterilized water was used for the control. Then, all pots were irrigated and covered for 3 days. Cucumber seeds were disinfected with sodium hypochlorite (NaCIO) 1% for 4 min and were planted in each pot. Germinated, not germinated seeds, and collapsed plants were recorded after 7 and 10 days for planting, and disease intensity was calculated as recommended [14]: 0 = no symptoms; 1 = seed rot, not germinated; 2 = brown rot on the stem base, plant is still standing; 3 = plant is wilted, laying on the ground; and 4 = plant is dead. DS was calculated from disease grades 0–3 using the following formula [15]:
where DS = disease severity, f = infection class frequencies, v = number of plants of each class, N = total of observed plants, and X = highest value of the evaluation scale.
\n
Cucumber seeds were surface-sterilized using 0.2% NaCIO for 2 min and rinsed several times with distilled water. Arbuscular mycorrhizal (AM) fungi were obtained from the Iraqi Ministry of Sciences and Technology’s laboratory. This mixture consists of propagated units of Glomus clarum (Nicol. Schenck) and Glomus mosseae (Nicol. Gerd) in a suspension form (1 × 106 unit L−1 concentration). Glomus spp. were identified and separated in two tubes by the experts at Iraqi Ministry of Sciences and Technology’s laboratory. Six healthy seeds of cucumber were planted in each pot (25 cm in diameter), which contained 3 kg of sterilized soil (clay:sand, 2:1, v/v) into each pot. For mycorrhizal inoculum, each pot was inoculated with dilution of 5 ml of either Glomus clarum or G. mosseae/L−1 water twice at the beginning of cultivation and after 14 days. As controls, the pots were provided with no AM + no pathogen, AM only, and pathogen only. For the pathogen inoculum, 5 ml of spore suspension (R. solani) was added at the beginning of cultivation. Six treatments were conducted as the following: Glomus clarum, G. mosseae, G. clarum + R. solani, G. mosseae + R. solani, control, and control + R. solani. Four replicates were made for each treatment. In this study, all plants did not receive any fertilizer and were watered when necessary at outdoor conditions. The disease severity for each treatment was monitored and estimated as mentioned above [16].
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When the plants emerged above the soil surface, five plants were harvested from each treatment after 5, 10, 15, and 20 days. The plants were washed with tap water to clean off soil particles. Fresh and dry weights were evaluated and recorded after drying the samples by a hot air oven at 60°C for 48 h until gaining constant weight [17].
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3. Results and discussion
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Five pathogens were isolated form the infected plants and soil. The fungal identification was performed according to the morphological characteristic as previously reported in literatures [18, 19]. Among five isolated pathogens, R. solani showed the highest disease severity (DS) on cucumber plants, which was about 63%, while Penicillium spp. showed the lowest disease severity (DS), which was about 8% (Figure 1). Therefore, R. solani was the most aggressive pathogen due to the suitable environment condition, and the availability of susceptible hosts and was used for all subsequent studies.
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Figure 1.
Pathogenicity test for isolated pathogens against damping-off diseases on cucumber. Each column represents the mean of five replicates. Bars on the pillars represent standard error and LSD = 5.73 (P = 0.01).
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The effect of AM fungi against R. solani on cucumber plants was studied by the inoculation of cucumber plants with the AM, G. mosseae + G. clarum, which showed a significant reduction in the disease severity of damping-off compared with control (Figure 2). Disease severity (DS) of mycorrhizal plants was reduced by 46% and 41%, respectively. Furthermore, inoculated plants with mycorrhiza showed fewer symptoms compared with non-mycorrhizal plants. Disease severity in AM-inoculated plants with G. mosseae was about 20%, which was slightly less than AM-inoculated plants with G. clarum (Figure 2).
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Figure 2.
Evaluation of arbuscular mycorrhizal (AM) fungi on the disease severity of damping-off diseases on cucumber. Each column represents the mean of four replicates. Bars on the pillars represent standard error and LSD (P = 0.01).
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The effect of AM fungi on the growth parameters of cucumber plants was assessed by shoot dry weight and root dry weight. AM fungi-colonized plants had significantly increased shoot and root dry weights when compared with the non-mycorrhizal plants (Table 1). Cucumber plants, colonized with AM (G. mosseae), showed a slight increase in all growth parameters compared with the plant colonized with AM (G. clarum), which matches with our results on the DS experiment (Table 1).
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\n
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\n
\n
\n
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\n
\n\n
\n
Treatment
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Shoot dry weight (g/plant)
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Root dry weight (g/plant)
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\n
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5 days
\n
10 days
\n
15 days
\n
20 days
\n
5 days
\n
10 days
\n
15 days
\n
20 days
\n
\n\n\n
\n
Control
\n
0.5
\n
0.8
\n
0.9
\n
1.1
\n
0.2
\n
0.4
\n
0.7
\n
0.9
\n
\n
\n
Control + R. solani\n
\n
0.1
\n
0.3
\n
0.4
\n
0.5
\n
0.08
\n
0.1
\n
0.2
\n
0.3
\n
\n
\n
\nGlomus clarum\n
\n
0.4
\n
0.6
\n
0.7
\n
1.2
\n
0.15
\n
0.3
\n
0.6
\n
0.8
\n
\n
\n
\nG. mosseae\n
\n
0.6
\n
0.7
\n
0.8
\n
1.1
\n
0.2
\n
0.4
\n
0.7
\n
0.9
\n
\n
\n
\nG. clarum + R. solani\n
\n
0.3
\n
0.5
\n
0.6
\n
0.9
\n
0.15
\n
0.2
\n
0.5
\n
0.6
\n
\n
\n
\nG. mosseae + R. solani\n
\n
0.4
\n
0.6
\n
0.8
\n
1
\n
0.2
\n
0.3
\n
0.6
\n
0.7
\n
\n\n
Table 1.
Evaluation of AM fungi on the growth parameters of cucumber plants.
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Mycorrhizal fungi are considered as ideal biocontrol agents due to some characteristics such as the ability to form a mutualistic symbiosis relationship with the roots of most vascular plant species [20]. Moreover, the plant-mycorrhiza relationship benefits the plant not only to control soil-borne pathogens but also to enhance the plant’s resistance to various abiotic stresses and increases the nutrients’ absorption [21].
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In the present study, inoculated plant with mycorrhizal fungi reduces significantly the disease severity of R. solani pathogen, which may be attributed to increase the nutrients’ status, reduce the direct competition for root space and resources with the pathogen, induce the plant’s immunity to involve certain systemic mechanisms such as the systemic acquired resistance (SAR) and cell wall defenses, and enhance the production of defense compounds such as phenolics, -1,3-glucanase, and chitinolytic enzymes [9]. Additionally, inoculated plants with mycorrhizal fungi (G. mosseae) showed a lower disease severity than G. clarum, which may lead to a potential active control tool. Furthermore, the inoculation with mycorrhizal fungi increases both the root dry weight and shoot dry weight, which supports our hypothesis.
\n
Mycorrhizal fungi play a main part in plant defense against pathogens and form a mutual relationship with plants. In summary, both mycorrhiza species could be an important tool to control soil-borne pathogens, increase plant nutrient absorption, and increase resistance to abiotic stresses. In future research, specific systemic mechanisms of mycorrhiza fungi against pathogens should be investigated more.
\n
\n
Acknowledgments
\n
The research was supported by University of Muthanna, Iraq. The authors acknowledge the Ministry of Sciences and Technology in Iraq for providing them with the isolates of arbuscular mycorrhizal (AM) fungi to complete their research.
\n
\n',keywords:"biological control, Rhizoctonia solani, arbuscular mycorrhiza, cucumber, damping-off diseases",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/73149.pdf",chapterXML:"https://mts.intechopen.com/source/xml/73149.xml",downloadPdfUrl:"/chapter/pdf-download/73149",previewPdfUrl:"/chapter/pdf-preview/73149",totalDownloads:91,totalViews:0,totalCrossrefCites:0,dateSubmitted:"June 24th 2020",dateReviewed:"July 2nd 2020",datePrePublished:"September 7th 2020",datePublished:null,dateFinished:null,readingETA:"0",abstract:"Rhizoctonia solani is one of the most important causative agents of damping-off diseases on cucumber plants and significantly reduces their yield. R. solani possesses some characteristics, such as wide host range and unlimited survival in soil, which made it most difficult to control. Therefore, the research for a biocontrol agent will be valuable to control this disease. Two species of mycorrhizal fungi (Glomus mosseae and Glomus clarum) that were evaluated against the agent R. solani reduced the damping-off disease on the cucumber plant. Mycorrhizal-inoculated plants with both species showed a significant reduction in disease severity (DS), which were 21 and 25%, respectively, whereas the disease severity was 65% for non-inoculated plants. Furthermore, the effects of mycorrhizal fungi were evaluated against the growth parameters of cucumber plants. Plants inoculated with both species of mycorrhizal fungi showed a significant increase in both shoot dry weight and root dry weight compared with uninoculated plants. In conclusion, both mycorrhiza species could be an important tool to control soil-borne pathogens, increase plant’s nutrients’ absorption, and increase resistance to abiotic stresses.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/73149",risUrl:"/chapter/ris/73149",signatures:"Baker Diwan Getheeth Aljawasim, Hussein M. 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The effectiveness of wood vinegar in controlling Rhizoctonia solani and Sclerotinia sclerotiorum in green house-cucumber. International Journal of Agricultural Sciences and Natural Resources. 2013;1(4):38-43\n'},{id:"B2",body:'\nBartz FE, Cubeta MA, Toda T, Naito S, Ivors KL. An in planta method for assessing the role of basidiospores in Rhizoctonia foliar disease of tomato. Plant Disease. 2010;94(5):515-520\n'},{id:"B3",body:'\nThakur M, Sahu NR, Tiwari P, Kotasthane A. Combination of azoxystrobin + difenocanazole provides effective management of sheath blight of rice caused by Rhizoctonia solani. IJCS. 2018;6(4):1682-1685\n'},{id:"B4",body:'\nVinale F, Sivasithamparam K, Ghisalberti EL, Marra R, Woo SL, Lorito M. Trichoderma–plant–pathogen interactions. Soil Biology and Biochemistry. 2008;40(1):1-10\n'},{id:"B5",body:'\nManganiello G, Sacco A, Ercolano MR, Vinale F, Lanzuise S, Pascale A, et al. Modulation of tomato response to Rhizoctonia solani by Trichoderma harzianum and its secondary metabolite harzianic acid. Frontiers in Microbiology. 2018;9:1966\n'},{id:"B6",body:'\nJustyna N, Magdalena S, Urszula M. Trichoderma atroviride enhances phenolic synthesis and cucumber protection against Rhizoctonia solani. Plant Protection Science. 2017;54(1):17-23\n'},{id:"B7",body:'\nVinale F, Marra R, Scala F, Ghisalberti E, Lorito M, Sivasithamparam K. Major secondary metabolites produced by two commercial Trichoderma strains active against different phytopathogens. Letters in Applied Microbiology. 2006;43(2):143-148\n'},{id:"B8",body:'\nLiu K, McInroy JA, Hu C-H, Kloepper JW. Mixtures of plant-growth-promoting Rhizobacteria enhance biological control of multiple plant diseases and plant-growth promotion in the presence of pathogens. Plant Disease. 2018;102(1):67-72\n'},{id:"B9",body:'\nJacott CN, Murray JD, Ridout CJ. Trade-offs in arbuscular mycorrhizal symbiosis: Disease resistance, growth responses and perspectives for crop breeding. Agronomy. 2017;7(4):75\n'},{id:"B10",body:'\nSmith SE, Smith FA, Jakobsen I. Mycorrhizal fungi can dominate phosphate supply to plants irrespective of growth responses. Plant Physiology. 2003;133(1):16-20\n'},{id:"B11",body:'\nHoeksema JD, Chaudhary VB, Gehring CA, Johnson NC, Karst J, Koide RT, et al. A meta-analysis of context-dependency in plant response to inoculation with mycorrhizal fungi. Ecology Letters. 2010;13(3):394-407\n'},{id:"B12",body:'\nDomsch KH, Gams W, Anderson T-H. Compendium of Soil Fungi. Vol. 1. London: Academic Press Ltd.; 1980\n'},{id:"B13",body:'\nJaiswal AK, Elad Y, Graber ER, Frenkel O. Rhizoctonia solani suppression and plant growth promotion in cucumber as affected by biochar pyrolysis temperature, feedstock and concentration. Soil Biology and Biochemistry. 2014;69:110-118\n'},{id:"B14",body:'\nKhan MR, Fischer S, Egan D, Doohan FM. Biological control of Fusarium seedling blight disease of wheat and barley. Phytopathology. 2006;96(4):386-394\n'},{id:"B15",body:'\nAbawi G, Widmer T. Impact of soil health management practices on soilborne pathogens, nematodes and root diseases of vegetable crops. Applied Soil Ecology. 2000;15(1):37-47\n'},{id:"B16",body:'\nAl-Askar A, Rashad Y. Arbuscular mycorrhizal fungi: A biocontrol agent against common. Plant Pathology Journal. 2010;9(1):31-38\n'},{id:"B17",body:'\nManila R, Nelson R. Nutrient uptake and promotion of growth by arbuscular mycorrhizal fungi in tomato and their role in bio-protection against the tomato wilt pathogen. Journal of Microbiology and Biotechnology Research. 2017;3(4):42-46\n'},{id:"B18",body:'\nSharma M, Gupta S, Sharma T. Characterization of variability in Rhizoctonia solani by using morphological and molecular markers. Journal of Phytopathology. 2005;153(7-8):449-456\n'},{id:"B19",body:'\nGuleria S, Aggarwal R, Thind T, Sharma T. Morphological and pathological variability in rice isolates of Rhizoctonia solani and molecular analysis of their genetic variability. Journal of Phytopathology. 2007;155(11-12):654-661\n'},{id:"B20",body:'\nSong Y, Chen D, Lu K, Sun Z, Zeng R. Enhanced tomato disease resistance primed by arbuscular mycorrhizal fungus. Frontiers in Plant Science. 2015;6:786\n'},{id:"B21",body:'\nSmith SE, Read DJ. Mycorrhizal Symbiosis. Academic Press; 2010\n'}],footnotes:[],contributors:[{corresp:"yes",contributorFullName:"Baker Diwan Getheeth Aljawasim",address:"baker.aljawasim@mu.edu.iq",affiliation:'
Department of Plant Protection, College of Agriculture, Al-Muthanna University, Iraq
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