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Revista mexicana de ciencias agrícolas
versão impressa ISSN 2007-0934
Rev. Mex. Cienc. Agríc vol.9 spe 20 Texcoco Abr./Mai. 2018
https://doi.org/10.29312/remexca.v0i20.1000
Articles
Antagonistic potential of bacteria and marine yeasts for the control of phytopathogenic fungi
1Center for Biological Research of the Northwest, SC. La Paz, Baja California Sur, Mexico. CP. 23096. Tel 01 (612) 1238484. (lhernandez@cibnor.mx; eltom-r@hotmail.com).
2Autonomous University of Baja California Sur. La Paz, Baja California Sur, Mexico. CP. 23080. Tel 01(612) 1238800. (mirerome22@hotmail.com; fruiz@uabcs.mx).
3Faculty of Agricultural Sciences-Campus Xalapa, Universidad Veracruzana. Xalapa, Veracruz, Mexico. CP. 91090. Tel. 01(228) 8421749 (cchiquito@uv.mx).
The application of synthetic fungicides is a common practice in the control of phytopathogenic fungi. However, its use indiscriminately has brought problems to human, animal and environmental health and has generated resistance in phytopathogens. In the search for alternatives, biological control using microorganisms can be an efficient option to the use of synthetic fungicides. Although bacteria and yeasts isolated from soil and plants have been evaluated as biological control agents, the search for new antagonists continues. The oceanic microflora can be an option for the selection of new antagonistic agents. The objective of this study was to evaluate the antagonistic potential of different bacteria (Stenotrophomonas rhizophila, Bacillus amyloliquefaciens and B. subtilis) and yeasts (Debaryomyces hansenii, Cryptococcus diffluens and Rhodotorula minuta) previously isolated from a hyperhialin lagoon against 13 phytopathogenic fungi of agronomic importance. The results show that different strains of the S. rhizophila bacterium were those that exerted greater inhibition on spore germination and mycelial growth of all the phytopathogenic fungi, surpassing the treatments with synthetic fungicides. Among the yeasts, the strains of D. hansenii stood out. According to their antagonistic capacity, marine microorganisms can be an option for the management of diseases caused by phytopathogenic fungi.
Keywords: biological control; synthetic fungicides; phytopathogenic fungi; marine microorganisms
La aplicación de fungicidas sintéticos es una práctica común en el control de hongos fitopatógenos. Sin embargo, su uso de manera indiscriminada ha traído problemas para la salud humana, animal, medio ambiente y ha generado resistencia en los fitopatógenos. En la búsqueda de alternativas, el control biológico usando microorganismos puede ser una opción eficiente al uso de fungicidas sintéticos. Aunque bacterias y levaduras aisladas de suelo y plantas han sido evaluadas como agentes de control biológico, la búsqueda de nuevos antagonistas continúa. La microflora oceánica puede ser una opción para la selección de nuevos agentes antagonistas. El objetivo de este estudio fue evaluar el potencial antagónico de diferentes bacterias (Stenotrophomonas rhizophila, Bacillus amyloliquefaciens y B. subtilis) y levaduras (Debaryomyces hansenii, Cryptococcus diffluens y Rhodotorula minuta) aisladas previamente de una laguna hiperhialina contra 13 hongos fitopatógenos de importancia agronómica. Los resultados muestran que diferentes cepas de la bacteria S. rhizophila fueron las que ejercieron mayor inhibición sobre la germinación de esporas y crecimiento micelial de todos los hongos fitopatógenos, superando a los tratamientos con fungicidas sintéticos. Dentro de las levaduras destacaron las cepas de D. hansenii. De acuerdo con su capacidad antagónica, los microorganismos marinos pueden ser una opción para el manejo de enfermedades ocasionadas por hongos fitopatógenos.
Palabras clave: control biológico; fungicidas sintéticos; hongos fitopatógenos; microorganismos marinos
Introduction
Worldwide, phytopathogenic fungi cause considerable economic losses due to the damage they cause to crops during the different stages of their development (eg. flowering, maturity, harvest) (Pusztahelyi et al., 2015; Mumford et al., 2016). Traditionally, its control has been based on the application of synthetic fungicides; however, the use of these products can cause harm to human, animal and ecosystem health (Tu et al., 2013; Moshi and Matoju, 2017), as well as generating resistance in phytopathogens (Liu et al., 2016; Romanazzi et al., 2016).
Biological control using antagonistic microorganisms may represent a viable and environmentally safe alternative to synthetic fungicides (Weaver et al., 2016; Bach et al., 2016). Bacteria and yeasts have been successfully used to control diseases (Eljounaidi et al., 2016; Wisniewski et al., 2016). Some of its main antagonistic mechanisms are competition for space and nutrients (Droby et al., 2016), inhibition by volatile organic compounds (CVO’s) (Raza et al., 2016a; Arrarte et al., 2017), siderophores (Sasha et al., 2016; Sasirekha and Srividya, 2016), antibiotics (Sharifazizi et al., 2017), hydrolytic enzymes (Ferraz et al., 2016; Tokpah et al., 2016), induction of resistance (Punja et al., 2016) among others.
Bacteria and yeasts are commonly isolated from the surfaces of plants and soil (Sharma et al., 2009; Larkin, 2016); however, there are other environments such as the ocean where microorganisms with antagonistic capacity could be isolated as efficient as synthetic fungicides. Studies in recent years on the oceanic microflora have been based mainly on describing its pharmaceutical properties, such as antimicrobial, antituberculous, antiviral, antiparasitic, anthelmintic, among others (Dewapriya and Kim, 2014; Jin et al., 2016). However, bacteria and yeasts from marine environments are being considered as new sources of products that can be applied in various areas such as agriculture because they have been shown to be highly efficient microorganisms in the biological control of phytopathogens (Wang et al., 2008; Hernández-Montiel et al., 2010; Wang et al., 2011; Medina-Córdova et al., 2016).
The antagonistic potential of marine microorganisms should be studied to select those as promising biological control agents that in the medium term can be a treatment that promote food security, ecological-sustainable production (Usall et al., 2016) and the development of new biological products (Vero et al., 2013). Therefore, the objective of this work was to evaluate the antagonistic potential of strains of bacteria and marine yeast against various phytopathogenic fungi of agronomic importance.
Materials and methods
Microorganisms used
The phytopathogenic fungi used in this study were Colletotrichum gloeosporioides, Penicillium italicum, P. digitatum, Alternaria solani, Fusarium oxysporum, Neoscytalidium dimidiatum A. alternata, F. solani y Curvularia sp. (Table 1), which belong to the Phytopathology Laboratory of the Center for Biological Research of the Northwest (CIBNOR) and the Autonomous University of Baja California Sur. The fungi were cultivated in Petri dishes with potato and dextrose agar culture medium (PDA) at 27 °C for 7 days. Their concentrations were adjusted to 1x104 spores mL-1. The collection of bacteria and marine yeasts were provided by the CIBNOR and were originally isolated from the Ojo de Liebre hyperhyaline lagoon, located between 27 °35’ and 27° 52’ north latitude and 113° 58’ and 114° 0’ of latitude west in the municipality of Mulege, Baja California Sur, Mexico.
Table 1 Origin of different phytopathogenic fungi of agricultural importance.
| Key£ | Phytopathogen | Hospedero | Disease |
| CIB-CGP | Colletotrichum gloeosporioides | Carica papaya L. | Anthracnose |
| CIB-CGM | C. gloeosporioides | Mangifera indica L. | Anthracnose |
| CIB-PIL | Penicillium italicum | Citrus aurantifolia (Christm.) Swingle | Blue rot |
| CIB-PDN | P. digitatum | Citrus sinensis (L.) Osbeck | Green rot |
| CIB-AST | Alternaria solani | Lycopersicon esculentum Mill. | Early blight |
| CIB-FOC | Fusarium oxysporum | Capsicum annuum L. | Root rot |
| CIB-FOA | F. oxysporum | Agave tequilana Weber | Root rot |
| MR-HF12 | Neoscytalidium dimidiatum | Ficus Carica L. | Descending death |
| MR-AA16 | A. alternata | Ocimum basilicum L. | Leaf spot |
| MR-FG16 | F. solani | Cicer arietinum L. | Root rot |
| MR-FE16 | F. oxysporum | Asparagus officinalis L. | Root rot |
| MR-FA16 | F. oxysporum | Ocimum basilicum L. | Root rot |
| MR-CP16 | Curvularia sp. | Washingtonia robusta Wendl. | Leaf spot |
£= fungi with the CIB key belong to the collection of the Center for Biological Research of the Northwest and with the key MR belong to the collection of the Autonomous University of Baja California Sur.
The bacteria selected were Stenotrophomonas rhizophila (strain KM01 and KM02), Bacillus amyloliquefaciens (strain RB01 and RB02) and B. subtilis (strain RBM01 and RBM02), which were cultivated in 250 mL Erlenmeyer flask with medium broth trypticasein (CST) at 25 °C for 24 h and 180 rpm. Selected yeasts were Debaryomyces hansenii (strain L01, L02 and L03), Cryptococcus diffluens (strain N02) and Rhodotorula minuta (strain R04 and R06), which were cultivated in 250 mL Erlenmeyer flask with half potato broth and dextrose (CPD) at 25 °C for 24 h and 180 rpm. Bacteria and yeast were used at a concentration of 1x106 cells mL-1.
Inhibition of spore germination of phytopathogenic fungi
In order to determine the antagonistic capacity of the microorganisms on the germination of spores of the different phytopathogens, the methodology proposed by Hernández-Montiel et al. (2010). One 1.5 mL Eppendorf tube, 500 μL of each bacterial or yeast suspension (both pre-adjusted to 1x106 cells mL-1) were combined with 500 μL of each fungus suspension (previously adjusted to 1x104 spores mL-1) and incubated 28 °C for 48 h. Another combination was made with 500 μL of each fungus with a synthetic fungicide, which was selected for each species of phytopathogen (Tecto 60 [ia 2-4-thiazolyl-1H-benzimidazole] at 5 g L-1 for Colletotrichum gloeosporioides, Penicillium italicum and P. digitatum, Amistar G. [ia Methyl E-2-2-6-2-cyanophenoxy pyrimidin-4-yloxy-phenyl-3-methoxyacrylate] at 3 g L-1 for Alternaria solani and A. alternata, Derosal 50 [ia Methyl-2-benzimidazol-carbamate] at 2 g L-1 for Fusarium oxysporum and F. solani and Cantus [ia 2-Chloro-N-4’-chlorobiphenyl-2-yl nicotinamide] at 1 g L-1 for Neoscytalidium dimidiatum and Curvularia sp.).
As a control, 500 μL of each fungus suspension was placed in a 1.5 mL Eppendorf tube. Aliquots of each treatment were taken to determine the number of whole and germinated spores, considering an entire spore as that which showed no change of color or rupture in its cell wall and a germinated spore when the size of hypha was equal to or greater than the diameter of the spore (Yao et al., 2004). Ten repetitions were made per treatment, observing 200 spores per repetition.
Inhibition of radial growth of phytopathogenic fungi
A 0.5 cm diameter PDA tampon containing the culture of each 7-day fungus was placed in the center of Petri dishes with PDA medium. Subsequently, 10 μL of each bacterial or yeast concentration was inoculated and striated at the two ends of the Petri dish. As a control, Petri dishes were inoculated only with the fungus. All Petri dishes were incubated at 25 °C for 7 days. At the end, the area of mycelial growth of each fungus was quantified using the ImajeJ® program and the (%) inhibition was determined by the formula I%= DC-DT/DCX100, where I%= inhibition of the fungus in percentage, DC= diameter of the mycelium of the control treatment and DT= diameter of the mycelium in the presence of the antagonist. Five repetitions were made per treatment.
Results and discussion
The germination of spores in all the phytopathogenic fungi was inhibited between 90 to 94% by strain KM01 of S. rhizophila, significantly surpassing (p< 0.05) the inhibition exerted by other marine microorganisms and synthetic fungicides (Table 2). Strains of B. amyloliquefaciens and B. subtilis inhibited between 81 to 93% and 51 to 69%, respectively. In relation to the yeasts, strains L01 and L02 of D. hansenii with a range of inhibition between 50 to 91% stood out. The lowest values of inhibition were observed with the strains of R. minuta and C. diffluens. The various fungicides used in this study inhibited spore germination by 80 to 90%. On the other hand, mycelial growth was inhibited in all fungi by strains KM01 and KM02 of S. rhizophila by 90 to 98% and 88 to 97%, respectively, significantly exceeding (p< 0.05) the inhibition exerted by the other marine microorganisms and synthetic fungicides (Table 3).
Table 2 In vitro effect of marine microorganisms on the inhibition of spore germination of phytopathogenic fungi.
| Cepa | Inhibition of spore germination (%) | ||||||||||||
| CIB-CGP | CIB-CGM | CIB-PIL | CIB-PDN | CIB-AST | CIB-FOC | CIB-FOA | MR-HF12 | MR-AA16 | MR-FG16 | MR-FE16 | MR-FA16 | MR-CP16 | |
| KM01 | 91.6a¥ | 93.5a | 94.2a | 94.7a | 90.2a | 91.4a | 91.8a | 91.8a | 92.3a | 90.5 a | 90.1 a | 91.4 a | 91.8a |
| KM02 | 90.8a | 83.6e | 90.6c | 94.6a | 74.6d | 75.6e | 86.7e | 81.6e | 80.2d | 84.7c | 81.9c | 90.7b | 90.1b |
| RB01 | 88.3b | 93.4a | 87.3e | 90.4c | 81.7b | 81.9d | 90.1b | 85.8d | 86.5b | 82.7d | 87.3b | 91.2a | 91.7a |
| RB02 | 83.9d | 85.6d | 91.8b | 88.9d | 81.9b | 87.4c | 87.7d | 89.7c | 85.6c | 81.1e | 87.5b | 84.9c | 89.1c |
| RBM01 | 60.4e | 69.4f | 57.6g | 60.2f | 58.9f | 58.1h | 54.6h | 66.1h | 62.5f | 63.7f | 66.5d | 55.7f | 59.5f |
| RBM02 | 60.2e | 61.7g | 54.7h | 57.4h | 58.6f | 54.9i | 58.4f | 57.3i | 51.9h | 51.8i | 59.1f | 60.1e | 61.7e |
| L01 | 89.1b | 90.1b | 91.7b | 91.7b | 69.5e | 64.7f | 52.6i | 65.8f | 66.2e | 57.7h | 54.1g | 62.7d | 58.1g |
| L02 | 89.3b | 87.9c | 87.1e | 86.3e | 49.7g | 54.7i | 53.7g | 53.8j | 53.1g | 50.9 j | 50.4h | 51.4g | 51.2h |
| L03 | 58.3f | 60.3h | 63.9f | 58.7g | 41.9h | 61.8g | 30.3j | 63.7g | 43.7i | 60.7 | 60.7e | 50.1h | 61.7e |
| N02 | 13.4h | 13.5k | 15.7i | 17.2i | 13.8i | 15.3j | 14.8k | 16.2k | 12.9k | 17.3k | 16.9i | 17.3i | 10.7k |
| R04 | 18.5g | 18.7j | 15.6i | 16.6j | 13.4i | 11k | 13.6l | 15.9k | 17.4 j | 13.2l | 15.3j | 13.4j | 15.8i |
| R06 | 12.9i | 19.9i | 13.2j | 10.7k | 13.6i | 6.9l | 9.8m | 10.9l | 11.7l | 11.7m | 9.7k | 12.7k | 11.8j |
| Fungicida | 85.4c§ | 90.2b§ | 89.6 d§ | 90.6c§ | 80.2c£ | 90.1b₡ | 87.8c₡ | 90.2b ℓ | 85.8c₺ | 88.1b₡ | 87.7b₡ | 90.3b₡ | 87.7d ℓ |
§= Tecto 60 (ia. 2-(4-tiazolil)-1H-bencimidazol) a 5 g L-1. ₺= Amistar G. (ia. Metil (E)-2-2-6-(2-cianofenoxi) pirimidin-4-iloxi-fenil-3-metoxiacrilato) a 3 g L-1; ₡= Derosal 50 (ia. Metil-2-bencimidazol-carbamato) a 2 g L-1. ℓ= Cantus (ia. 2-Cloro-N-(4’-clorobifenil-2-il) nicotinamida) a 1 g L-1. ¥= different letters in the columns indicate significant differences according to the Fisher LSD post-hoc test (p <0.05).
Table 3 In vitro effect of marine microorganisms on the inhibition of radial growth of phytopathogenic fungi.
| Cepa | Inhibition of radial growth (%) | ||||||||||||
| CIB-CGP | CIB-CGM | CIB-PIL | CIB-PDN | CIB-AST | CIB-FOC | CIB-FOA | MR-HF12 | MR-AA16 | MR-FG16 | MR-FE16 | MR-FA16 | MR-CP16 | |
| KM01 | 97.2a¥ | 98.4 | 97.1a | 96.7a | 93.6a | 95.4a | 90.7a | 93.1a | 93.2a | 90.6a | 91.1a | 93.2a | 95.9a |
| KM02 | 97.4a | 94.7b | 94.5b | 96.3a | 87.5b | 93.2b | 90.5a | 90.5b | 88.1b | 90.5a | 90.9a | 91.8b | 93.7b |
| RB01 | 91.5b | 83.1d | 89.3d | 88.2d | 84.1c | 85.1e | 82.1c | 84.3d | 86.7c | 83.1c | 86.5c | 86.3d | 85.1d |
| RB02 | 91.1b | 83.4d | 89.7d | 92.5b | 87.3b | 86.9d | 82.6c | 84.4d | 83.2d | 82.7c | 83.2d | 80.1e | 85.4d |
| RBM01 | 64.3d | 57.4f | 73.4e | 61.4e | 75.4e | 50.7g | 56.7e | 58.2f | 71.3e | 55.6e | 54.7f | 61.7g | 71.3e |
| RBM02 | 64.7d | 61.7e | 59.6f | 58.3f | 74.1f | 60.6f | 68.3d | 68.7e | 64.1f | 60.7d | 63.3e | 68.1f | 62.8f |
| L01 | 15.5f | 19.3g | 13.4h | 15.2i | 12.7i | 13.1i | 18.7f | 14.3g | 12.8h | 15.1g | 17.4g | 18.3h | 15.3h |
| L02 | 15.2f | 14.2i | 13.2h | 18.7h | 20.2g | 18.1h | 10.8h | 14.4g | 19.6g | 15.4g | 16.2h | 14.5i | 17.4g |
| L03 | 18.1e | 18.4h | 20.7g | 19.2g | 15.6h | 18.5h | 15.1g | 10.1h | 11.1i | 17.3f | 16.5h | 18.3h | 17.7g |
| N02 | 10.7g | 11.8j | 12.1i | 8.1j | 9.1j | 9.4j | 14.9g | 8.5i | 11.3i | 11.3h | 10.1i | 11.7j | 10.8i |
| R04 | 6.1h | 5.2k | 4.3k | 7.8k | 5.1l | 6.8k | 5.6i | 5.1k | 6.2j | 5.8i | 7.8j | 7.6k | 6.6j |
| R06 | 5.7i | 4.7l | 7.1j | 7.6k | 6.2k | 9.7j | 5.7i | 6.2j | 4.9k | 6.1i | 6.3k | 7.4k | 5.1k |
| Fungicida | 86.1c§ | 91.1c§ | 90.3c§ | 89.7c§ | 81.5d£ | 90.7c₡ | 89.3b₡ | 86.6c ℓ | 86.3c₺ | 88.9b₡ | 88.1b₡ | 90.1c₡ | 90.2c ℓ |
§= Tecto 60 (ia. 2-(4-tiazolil)-1H-bencimidazol) a 5 g L-1; ₺= Amistar G. (ia. Metil (E)-2-2-6-(2-cianofenoxi) pirimidin-4-iloxi-fenil-3-metoxiacrilato) a 3 g L-1. ₡ = Derosal 50 (ia. Metil-2-bencimidazol-carbamato) a 2 g L-1. ℓ = Cantus (ia. 2-Cloro-N-(4’-clorobifenil-2-il) nicotinamida) a 1 g L-1; ¥ = different letters in the columns indicate significant differences according to Fisher’s LSD post-hoc test (p <0.05).
Strains of B. amyloliquefaciens and B. subtilis inhibited between 80 to 92% and 50 to 75%, respectively. In relation to the yeasts, the strains of D. hansenii presented an inhibition of 10 to 20%, the lowest values were observed with the strains of R. minuta. The various fungicides inhibited between 86 to 91%. This ability to inhibit spore germination and the mycelial growth of phytopathogenic fungi by bacteria and yeasts has already been studied in isolated plant or soil strains (Mnif and Ghribi, 2015; Kröber et al., 2016; Palazzini et al., 2016; Reiss and Jørgensen, 2017).
Among the main antagonistic mechanisms of bacteria and yeasts, there is the production of hydrolytic enzymes, competition for space and nutrients and siderophores (Droby et al., 1989; Kai et al., 2007; Ryan et al., 2009; Herzog et al., 2016; Medina-Córdova et al., 2016; Grzegorczyk et al., 2017). In relation to hydrolytic enzymes (chitinases, glucanases and proteases), these present an activity directly on the cell wall of the fungus, which, is composed mainly of chitin, β-glucan and proteins, which are hydrolyzed to produce oligosaccharides from smaller size that are harnessed as carbon by bacteria and yeast (Sharma et al., 2009).
The competition for nutrients and space is another antagonistic mechanism presented by microorganisms (Jamalizadeh et al., 2011) and is directly related to carbon competition in the environment, which is rapidly diminished by bacteria and yeast, limiting the fungus in its host germination and infection processes (Janisiewicz and Korsten, 2002; Liu et al., 2013).
On the other hand, the siderophores produced by bacteria or yeasts are molecules of low molecular weight related to the Fe3+ ion, which is trapped and transported by the microorganisms in an active transport process, using a multitude of membrane receptors. Once inside the cell, the iron is released through a redox process. Without iron in the environment, microorganisms cannot continue with their vital biological processes such as the synthesis and repair of nucleic acids, respiration, photosynthetic transport, nitrate reduction, free radical detoxification, among others. This strategy of siderophore production by bacteria and yeast has been involved in the control of phytopathogens and has been recognized as an important antagonist trait found in many of the biological control agents (Yu et al., 2011; Sasha et al., 2016; Liu et al., 2017).
In relation to the in vitro effect of COV’s, the inhibition exerted by the marine strains KM01 and KM02, both of S. rhizophila, towards all the phytopathogenic fungi was from 92 to 95%, significantly exceeding (p< 0.05) the inhibition exerted by other marine microorganisms (Table 4). The strains of B. amyloliquefaciens and B. subtilis inhibited between 81 to 89% and 60 to 69%, respectively. In relation to the yeasts, the strains of D. hansenii presented a range of inhibition between 44 to 59%. The lowest inhibition values were observed with R. minuta strains. The production of COV’s has already been identified as a way to inhibit spore germination and mycelial growth of fungi (Raza et al., 2016a; Arrarte et al., 2017).
Table 4 In vitro effect of marine microorganisms on the inhibition of radial growth of phytopathogenic fungi by COV’s.
| Cepa | Inhibition of growth (%) | ||||||||||||
| CIB-CGP | CIB-CGM | CIB-PIL | CIB-PDN | CIB-AST | CIB-FOC | CIB-FOA | MR-HF12 | MR-AA16 | MR-FG16 | MR-FE16 | MR-FA16 | MR-CP16 | |
| KM01 | 94.1a¥ | 92.2a | 95.4a | 93.6a | 96.1a | 93.1a | 94.9a | 94.4a | 94.8a | 91.5a | 95.5a | 95.2a | 93.2a |
| KM02 | 93.8a | 91.8b | 93.9b | 92.9b | 94.7b | 92.7b | 94.4a | 93.8b | 92.6b | 91.5a | 95.9a | 94.8a | 93.1a |
| RB01 | 83.4b | 85.4c | 87.8c | 85.6d | 84.1c | 89.1c | 83.5c | 84.8d | 86.1c | 80.2c | 82.4c | 83.4c | 89.4b |
| RB02 | 81.1c | 81.6d | 85.6d | 87.8c | 84.2c | 83.9d | 85.6b | 87.1c | 83.8d | 84.5b | 88.1b | 87.6b | 87.5c |
| RBM01 | 61.9d | 68.7e | 69.4f | 63.4f | 68.4d | 69.7e | 66.4d | 69.1e | 63.5e | 67.1d | 64.7d | 63.8e | 66.3d |
| RBM02 | 60.8e | 60.1f | 70.1e | 68.7e | 65.5e | 65.8f | 62.9e | 67.5f | 61.5f | 63.4e | 64.1d | 67.4d | 63.1e |
| L01 | 52.9g | 48.5h | 51.7h | 57.2g | 54.4f | 52.7g | 51.2g | 58.2g | 54.4h | 56.8f | 55.7f | 54.9g | 56.1f |
| L02 | 56.6f | 49.1g | 54.3g | 53.6h | 52 g | 52.9g | 59.5f | 55.3h | 57.5g | 54.1g | 58.9e | 57.2f | 54.7g |
| L03 | 50.1h | 49.3g | 47.5i | 51.2i | 50.3h | 49.8h | 48.3h | 48.7i | 45.6i | 48.5h | 44.1g | 49.7 h | 45.7h |
| N02 | 20.4i | 22.6i | 21.5j | 23.6j | 20.8i | 20.7i | 26.9i | 22.4j | 21.5j | 19.5i | 17.4h | 20.5i | 22.8i |
| R04 | 10.1j | 10.6j | 9.1l | 9.5k | 8.7k | 10.1j | 9.9j | 9.3k | 8.5k | 9.8j | 7.9i | 9.5j | 8.4k |
| R06 | 9.8j | 10.4j | 10.7k | 9.7k | 9.9j | 9.8j | 10.4j | 10.1l | 8.9k | 9.7k | 8.2i | 9.6j | 10.4j |
| Testigo | 0 k | 0 k | 0 m | 0 l | 0 l | 0 k | 0 k | 0 m | 0 l | 0 l | 0 j | 0 k | 0 l |
¥= different letters in the columns indicate significant differences according to Fisher's LSD post-hoc test (p< 0.05).
The COV’s produced by bacteria such as dimethyl disulfide, dimethylhexadecylamine, phenylethyl alcohol, furan 2-methyl-5-methyl, among others, and those produced by yeasts such as 2-methyl-1-propanol, 3-methyl-1-butanol, 2-methyl-1-butanol, among others, have already been reported in the inhibition of phytopathogens (Hernández-León et al., 2015; Raza et al., 2016b). In general, the antimicrobial activity of these compounds is attributed to their interaction with the cell membrane of the phytopathogen, which breaks down the acceleration of the diffusion of their ions and essential metabolites of their membrane (Heipieper et al., 1994).
Finally, the management of diseases caused by phytopathogenic fungi in plants through antagonistic microorganisms is a priority worldwide (Bardin et al., 2015; Stenberg et al., 2015; Van Bruggen and Finckh, 2016). This is the first study to demonstrate the antagonistic potential of marine bacteria of the species Stenotrophomonas rhizophila, Bacillus amyloliquefaciens and B. subtilis and of marine yeasts of the species Debaryomyces hansenii, Cryptococcus diffluens and Rhodotorula minuta towards different phytopathogenic fungi of soil and plants, surpassing the inhibition of the different synthetic fungicides used in this work.
Subsequent studies will study the antagonistic mechanisms (e.j competition space and nutrients, hydrolytic enzymes, siderophores, among others) of the best marine strains of bacteria and yeast, in addition to determine their ability to control diseases caused by fungi in vivo. The selection of the best marine microorganisms as antagonistic agents can be an alternative in the production of food in a sustainable way, reducing the dependence on synthetic fungicides and lowering the production costs of crops.
Conclusions
The greater antagonistic capacity of the different marine strains of bacteria and yeasts towards the different phytopathogenic fungi was observed with the KM01 strain of the S. rhizophila bacterium, which inhibited spore germination between 90 and 94% and between 90 and 94%. 98% mycelial growth of the fungi Colletotrichum gloeosporioides, Penicillium italicum, P. digitatum, Alternaria solani, Fusarium oxysporum, Neoscytalidium dimidiatum, A. alternata, F. solani y Curvularia sp., surpassing the effect of synthetic fungicides. Among the marine yeasts, strains L01, L02 and L03 of D. hansenii stood out. The antagonistic efficiency of marine microorganisms suggests that they may be a medium-term option in the management of diseases caused by phytopathogenic fungi.
Gratefulness
The authors thank the financial support of the National Council of Science and Technology (CONACYT) through project SEP-2012-181972 and Geol. Ernesto Díaz Rivera for his excellent technical assistance.
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Received: December 2017; Accepted: January 2018
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