Abstract
An indigenous microbial consortium was developed in order to combat late blight disease of tomato. For obtaining better insight in the antagonistic potential of native bioagents, 33 isolates of bioagents were isolated and evaluated against Phytophthora infestans causing late blight of tomato. Upon in-vitro screening of the varied isolates, the highest growth inhibition of the pathogen was recorded in Pseudomonas isolates; Pf-2 (81.33%) and Pf-3 (73.33%) followed by Trichoderma isolates; T-11 (73.73%) and T-14 (66.67%). All potent native microbial isolates showed consistent ability to produce siderophore, ammonia, IAA, HCN, volatile metabolites and also able to release inorganic phosphorus from tri-calcium phosphate. The potential isolates were identified as T. asperellum and P. fluorescens based on the molecular characterization. In-vitro compatibility analysis of microbial consortia showed positive interaction. The potent biocontrol consortial sets of Trichoderma and Pseudomonas were tested in-vitro and highest inhibition of the pathogen was recorded in the combination of Pf-2 + Pf-3 + T-11 + T-14 (83.33%) followed by Pf-2 + Pf-3 + T-11 (78.38%). Liquid bio-formulations were prepared using the best two microbial consortia (MC) which were utilized for the management of late blight through seed treatment (1%), soil application (1%) and foliar spray (1%) under natural epiphytotic conditions. The highest reduction of late blight severity was recorded in chemical control treatment (91.92%) followed by MC-1 (84.38%) and MC-2 (77.20%). The MC-1 also significantly promoted the tomato plant height (101.20%), number of leaves per plant (116.48%), number of branches per plant (146.57%), number of fruits per plant (185.52%), fresh weight of fruit (42.59%), root length (67.28%) and marketable fruit yield (313.02%) over control treatment whereas chemical treatment showed non-significant with all above parameters. Among the tested microbial consortia, outstanding results were obtained in MC-1 indicating better plant growth promoting potential and disease reduction potential and thus exhibiting tremendous potential for its commercial exploitation.
Similar content being viewed by others
Explore related subjects
Discover the latest articles, news and stories from top researchers in related subjects.Avoid common mistakes on your manuscript.
Introduction
Tomato is an indispensable vegetable crop which is the major source of nutrients and medicinal values, hence known as ‘Nutraceutical vegetable’ (Singh et al. 2019). Tomato is highly adaptive to warm season and can be grown successfully in plains as well as in hills. Cultivation of tomato in rainy season is assuming a great importance in the north-eastern region of India in general and Nagaland in particular owing to its high prices of produce obtained from other parts of the country during this period (Babu 2006).
Though tomato crop occupies a very important place among the vegetable crops cultivated in India, the average yield of this crop on farmers’ fields is reasonably poor. One of the constrain for poor yield is the devastating effect of certain diseases. Among the diseases, late blight of tomato caused by Phytophthora infestans is destructive and wide spread in nature (Son et al. 2008). Worldwide losses were estimated is about $170 billion annually and thus this pathogen was considered as a major threat for global food security (Latijnhouwers et al. 2004; Wu et al. 2012). Yield losses up to 79% from late blight damage in tomato have been recorded in India (Arora et al. 2014; Chowdappa et al. 2015).
Although, several management components viz., cultural practices, biological agents, host resistance and fungicides are available, but fungicides hold promise in managing the late blight disease of tomato. Use of fungicides is costly, may lead to environmental pollution and less effective due to increasing resistance of the pathogen. Under such conditions, the most effective method is the biological control (Harish et al. 2008). In recent years, emerging strategy is integrated biological control as microbial consortia. Under field conditions microbial consortia are much more efficient than single strain of organisms with diverse metabolic capabilities (Yan et al. 2002).
Hence, looking into the aforesaid realities, the use of native biological agents as a consortium and also not much systematic research work has been carried out on late blight disease of tomato under Nagaland condition. Hence, in the present study attempts were made to explore native isolates of BCAs and developing an indigenous microbial consortium package for developing a biointensive management strategy against late blight disease of tomato along with yield enhancement.
Materials and methods
Identification of the pathogen
The causal agent of late blight disease of tomato was isolated by standard tissue isolation technique on rye-A agar medium (Hollomon 1965). The purified isolate was subjected to pathogenicity test. For this purpose, isolated pathogen was inoculated on 4 weeks old susceptible tomato cv. Pusa Ruby (Loliam et al. 2012).
Morphological characters of the pathogen was studied on host as well as in pure culture on rye-B agar medium. The isolated pathogen was identified on the basis of morphological characters as documented by Waterhouse (1963).
Isolation and identification of bioagents
A field survey was undertaken for the collection of rhizospheric soil samples from different cropping areas in Nagaland, India (Table 1). Soil samples were taken from the rhizosphere of healthy plants and kept in polyethylene bags. The individual sample was mixed thoroughly after air drying. Thirty three isolates were obtained from the collected samples by soil dilution plate technique (Waksman 1927).
Initially isolated microbes were identified as Trichoderma spp. and Pseudomonas spp. based on morphological characteristics by use of selective media viz., Trichoderma selective medium (TSM) (Elad and Chet 1983) and King’s B medium (King et al. 1954) respectively. Further, the potential isolates were identified as T. asperellum (T-11; Acc. No MK928414 and T-14; Acc. No MK928417) and P. fluorescens (Pf-2; Acc. No MN783298 and Pf-3; Acc. No MN783297) based on the molecular characterization (Singh et al. 2020).
In-vitro antagonistic tests
The antagonistic effect of Trichoderma and Pseudomonas isolates were evaluated against Phytophthora infestans by dual culture plate technique as per Sivakumar et al. (2000) and Georgakopoulos et al. (2002) respectively. Linear mycelial growth of the pathogen was recorded in Petri plate after full growth of pathogen attained in control treatment. The per cent inhibition of the growth of pathogen by antagonists over control was calculated (Vincent 1927).
Investigation on the biocontrol mechanisms of antagonists
The effects of volatile metabolites and mycoparasitism activity of isolated BCAs were assessed against P. infestans by adopting the technique given by Dennis and Webster (1971) and Rodrigues (2010), respectively. The production of Ammonia, IAA and HCN by Trichoderma and Pseudomonas isolates were also determined in the qualitative assay technique given by Cappuccino and Sherman (1992), Gordon and Weber (1951) and Miller and Higgins (1970), respectively. Phosphate solubilization and siderophore production test was also conducted qualitatively by inoculation of Trichoderma and Pseudomonas isolates on National Botanical Research Institute’s phosphate (NBRIP) agar medium (Nautiyal 1999) and chrome azurol sulfonate (CAS) agar medium (Milagres et al. 1999), respectively.
Selection of potential isolates and their compatibility study in-vitro
Based on in-vitro antagonistic capabilities of Trichoderma and Pseudomonas isolates against P. infestans and elucidation of their various biocontrol mechanisms, the potent isolates were selected for further studies. In-vitro compatibility test amongst microbial consortia of potent isolates of Trichoderma and Pseudomonas were evaluated by dual culture plate method (Siddiqui and Shaukat 2003) in order to determine the compatibility among different combination of consortia.
Antagonistic efficacy of microbial consortia against P. infestans
The in-vitro bioassay technique was used for the testing of microbial consortia against P. infestans. The mycelial disc (10 mm diameter) of the pathogen (9 days old) was placed at centre of Petri plate containing rye-B agar medium (20 ml). Simultaneously, 10 mm diameter disc of potent Trichoderma (T-11 and T-14) isolates (9 days old) and 20 µl of an overnight culture of potent Pseudomonas (Pf-2 and Pf-3) isolates were poured in wells (5 mm diameter) at different corner of Petri plate. Linear mycelial growth of the pathogen was recorded in Petri plate when mycelium of test pathogen touched any antagonists in any treatment. The per cent inhibition of the growth of pathogen by antagonists over control was calculated (Vincent 1927).
Preparation of liquid based bio-formulation of microbial consortia
The conidial suspension of each selected isolates of T. asperellum (T-11 and T-14) was prepared from 9 days old PDA plates. The plates were rinsed with sterile distilled water and the mycelia were carefully scraped off with a bent glass rod. This suspension was filtered through filter paper (Whatman No.1) to separate the spores from the mycelia. The spore concentration was adjusted to 3.7 × 108 spores/ml (Dubos 1987) with the help of haemocytometer. Similarly, selected P. fluorescens isolates (Pf-2 and Pf-3) cell suspension was prepared by inoculating into King’s B broth followed by shaking for 48 h (150 rpm) at 28 °C. The bacterial suspension was adjusted optically at 1 × 109 cfu/ml (Mulya et al. 1996). Liquid based bio-formulations of consortia were prepared by mixing equal volume of each selected isolate just before use for field experiment (Srinivasan and Mathivanan 2009).
Field evaluation of liquid bio-formulation of microbial consortia against late blight of tomato under natural epiphytotic conditions
The field trials were conducted during the tomato growing seasons (Sept.–Jan.) of 2017–2018 and 2018–2019. The research field site is located in the foothills of Nagaland (India) and situated at 25° 45′ 45″ North latitude and 93° 51′ 45″ East longitudes at an elevation of 310 m above mean sea level.
The bio-formulation of microbial consortia (MC) and chemical treated seeds (400 seeds/treatment) of tomato were sown in nursery beds (8 × 1 cm at 1 cm depth) after 15 days of formalin (2%) treated soil. The tomato cv. Pusa Ruby was used in the field experiment, which is known to be highly susceptible to P. infestans in India (Singh et al. 2019). The 28 days old seedlings were transplanted (60 × 45 cm) in main field during second week of October in raised plot (1.8 m × 1.8 m). The each plot was framed at 50 cm distance apart. All the recommended standard cultural operations were followed.
The field experiment was laid out in a randomized block design (RBD) with six replications (72 plants per treatment). A total of four treatments viz., T1 (MC-1; seed treatment (1%) + soil application (1%) + foliar sprays (1%) at 15, 30 and 45 DAT), T2 (MC-2; seed treatment (1%) + soil application (1%) + foliar sprays (1%) at 15, 30 and 45 DAT), T3 (Chemical control; seed dressing with 0.3% Captan 50% WP + soil application of Mancozeb 75% WP (0.2%) + foliar sprays of Ridomil MZ 72% WP (0.25%) at 15, 30 and 45 DAT) and T4 (Control, sterile distilled water) were used.
Application methods of liquid microbial consortia
Seed treatment
The surface sterilized (1.0% sodium hypochlorite for 2 min) seeds were soaked in conidial suspension of microbial consortia at 1%, chemical control treatment (0.3% of captan 50% WP) and control treatment (soaked in sterile distilled water). All the treated seeds were dried by keeping under aseptic condition in laminar air flow for 5 h (Srinivasan and Mathivanan 2009).
Soil application
The soil application treatment was done with 1% of MC inoculated in FYM, Mancozeb 75% WP at 0.2% and sterile distilled water for control treatment at 10 days before transplanting (Srinivasan and Mathivanan 2011).
Foliar spray
Three foliar sprays were done with 1% of MC, Mancozeb + Metalaxyl-72% WP at 0.25% and sterile distilled water for control treatment at 15, 30 and 45 DAT. The total spray solution of 150 ml was used in each plot (12 plants) (Srinivasan et al. 2009).
Observations
The late blight disease severity was assessed visually on leaves, stems and fruits of all plants in each replication following rating scale as per Irzhansky and Cohen (2006), when all plants in control treatment infested with late blight disease under natural epiphytotic conditions. The severity grades were converted into percentage disease index (PDI) for analysis as per the formula given by Wheeler (1969).
Plant growth promoting attributes like plant height, number of leaves, number of branches, number of fruits per plant, fresh weight of fruit, marketable fruit yield and root length were recorded.
Statistical analysis
The data were analyzed using WASP 2.0 software developed by the Central Coastal Agricultural Research Institute, Goa (India).
Results
Identification of the pathogen
The pure culture was obtained from the diseased specimens were identified as P. infestans based on macroscopic and microscopic characters. The phenotypic characteristics of isolate were observed fluffy cottony mycelium and slow growth rate on the rye-B agar medium. Microscopic observation revealed that the fungal hyphae were hyaline, moderately thick hyphae, coenocytic and profusely branched. Sporangiosphores were sympodial with a small swelling at the base of each branch. Sporangia were terminal or lateral, ellipsoid, ovoid or limoniform, semipapillate, deciduous and pedicelless and they comparatively more frequently observed on the tomato plants than in pure culture. Chlamydospores of the pathogen were also recorded in diseased specimens (Fig. 1).
In-vitro antagonistic tests
Altogether 25 isolates of Trichoderma were screened for their inhibitory action on the radial growth of P. infestans. It was found that the growth of the pathogen in dual culture plates progressed until they come in contact with the leading edges of the antagonist. The per cent inhibition over control was calculated and it was observed that T-11 was the most promising isolate against P. infestans with 73.73 per cent inhibition. Next best isolate was T-14 (66.67%) followed by T-5 (64.93%), T-25 (64.00%) and the least antagonistic effect was observed in T-17 (51.07%) at 8 days after incubation at 18 ± 1 °C (Table 2).
The antagonistic effects of Pseudomonas isolates were evaluated against P. infestans which significantly inhibited the growth of the pathogen as compared to control treatment. Among the Pseudomonas isolates, maximum per cent inhibition was observed in Pf-2 (81.33%) which is significantly superior to all other treatments followed by Pf-3 (73.33%), Pf-1 (69.33%), Pf-7 (66.67%) and Pf-4 (65.33%) at 8 days after incubation at 18 ± 1 °C (Table 2). The clear zone of inhibition was also observed in the dual culture plate of Pf-2 and Pf-3.
Investigation on the biocontrol mechanisms of antagonists
The effects of volatile metabolites of Trichoderma and Pseudomonas isolates were assessed against P. infestans. Among the tested isolates, the per cent inhibition over control was calculated and it was recorded that Trichoderma (T-11) and Pseudomonas (Pf-3) was found to be most promising in production of volatile compounds against P. infestans with 45.55 and 53.67 per cent inhibition (Table 2). The mycoparasitism activity of 25 isolates of Trichoderma were also assessed against P. infestans and they showed the presence of coiling as hyphal interactions between them (Table 2).
The production of Ammonia by Trichoderma and Pseudomonas isolates were also determined in the qualitative assay. Among the tested isolates, Pseudomonas isolates (Pf-3, Pf-4, Pf-7 and Pf-8) and Trichoderma isolates (T-1, T-2, T-11, T-14 and T-25) exhibited strong ammonia production by turning initial peptone water broth from yellow to dark brown colour (Table 2). The results of qualitative assay of IAA production by different native BCAs revealed that Pseudomonas isolates (Pf-2 and Pf-3) and Trichoderma isolates (T-11 and T-14) exhibited strong IAA production. The moderate production of HCN was observed in Pseudomonas isolates (Pf-2, Pf-3 and Pf-8) (Table 2).
The results of qualitative assay of phosphate solubilization by different native BCAs revealed that Pseudomonas isolates (Pf-2, Pf-3 and Pf-8) and Trichoderma isolates (T-11 and T-14) elucidated strong phosphate solubility activity (Table 2). The siderophore production test was also conducted qualitatively by inoculation of Trichoderma and Pseudomonas isolates on chrome azurol sulfonate (CAS) agar medium. All 33 isolates showed positive results for siderophore production. Among the tested isolates, Pseudomonas isolates (Pf-3 and Pf-8) and Trichoderma isolates (T-3, T-4, T-5, T-7, T-8, T-9, T-10, T-11, T-14, T-15, T-18 and T-21) exhibited strong siderophore production by pink and orange halo colour development (Table 2).
Selection of potential isolates and their compatibility study in-vitro
Based on in-vitro antagonistic capabilities of Trichoderma and Pseudomonas isolates against P. infestans and elucidation of their various biocontrol mechanisms, the potent isolates of Pseudomonas (Pf-2 and Pf-3) and Trichoderma (T-11 and T-14) were used for further studies. Selected native microbial isolates were showed consistent ability to produce siderophore, ammonia, IAA, volatile metabolites and also able to release inorganic phosphorus from tri-calcium phosphate (Table 2).
In-vitro experiment was carried out in all permutations and combination amongst the potent isolates of Trichoderma and Pseudomonas. Altogether 11 treatment combinations were tested and compared with growth of Pf-2 (Control-1), Pf-3 (Control-2), T-11 (Control-3) and T-14 (Control-4). The microorganisms showing positive compatibility among them was recorded, tabulated and selected for further study. The data showed compatibility among all the treatment combinations of the four bioactive microorganisms in-vitro. No clear inhibition zone was recorded between the tested microbial consortia. Absence of inhibition zone indicated that the potential isolates of Trichoderma and Pseudomonas were compatible with each other.
Antagonistic efficacy of microbial consortia against P. infestans
A total of 12 treatment combinations were compared. Eleven consortia produced varying inhibitions (%) in-vitro against P. infestans (Table 3 and Fig. 2). All consortia tested against P. infestans were significantly superior over control. Among the different consortial sets tested in-vitro the significant highest inhibition of pathogen was recorded in the combination of Pf-2 + Pf-3 + T-11 + T-14 (83.33%) followed by Pf-2 + Pf-3 + T-11 (78.38%), Pf-2 + Pf-3 (77.43%) and Pf-2 + T-14 (76.54%) respectively at 5 days after incubation at 18 ± 1 °C.
An in-vitro study was taken up to select the two best microbial consortia (MC) against the test pathogen. The MC-1 (P. fluorescens Pf-2 + P. fluorescens Pf-3 + T. asperellum T-11 + T. asperellum T-14) and MC-2 (P. fluorescens Pf-2 + P. fluorescens Pf-3 + T. asperellum T-11) inhibited the pathogen significantly and were found to be the most effective consortia. Hence, they were selected for field study.
Field evaluation of liquid bio-formulation of microbial consortia against late blight of tomato under natural epiphytotic conditions
Among different treatments, liquid microbial consortia (MC)-1 significantly decreased the late blight severity (12.08 PDI) compared to all other treatments. This was comparable with the chemical treatment (6.25 PDI). In case of untreated control, high severity of 77.36 PDI was recorded at 45 DAT (Table 4). These results revealed that the chemical control significantly decreased late blight severity (91.92%) over control treatment. Next in order of merit was MC-1 (84.38%) and MC-2 (77.20%). The mortality per cent significantly decreased in all the treatment (100%) over control treatment (Table 4). Simultaneously, liquid bio-formulation of MC-1 significantly increased the tomato plant height (101.20% and Fig. 3), number of leaves per plant (116.48%), number of branches per plant (146.57%), number of fruits per plant (185.52%), fresh weight of fruit (42.59%), root length (67.28%) and marketable fruit yield (313.02%) over untreated control (Tables 5 and 6).
Discussions
The pathogen was identified as P. infestans based on the nature of disease observed, morphological and cultural characters seen under the microscope. These characters were further compared with the characters reported by Waterhouse (1963) and description given by Agrios (1997) and Zentmyer (1983). The present observations corroborates with the descriptions and findings of earlier workers.
The development of native bio-formulation is more efficient antagonistic player in plant disease management and growth promotion. Antagonistic effect of Trichoderma and Pseudomonas isolates against P. infestans has already been reported by several research workers (Kabir et al. 2013; Kumar et al. 2015; Lamsal et al. 2013; Patel and Mukadam 2011; Zegeye et al. 2011). In this study, the probable reasons of high inhibitory activity of the antagonists observed on P. infestans in dual cultures may be due to production of antifungal metabolites such as mycoparasitic activities, volatile gases, cell-wall degrading enzymes, HCN, siderophores, pyoluteorin, pyrrolnitrin and 2-4 diacetyl phloroglucinol.
Many secondary metabolites have been recorded to be involved in microbial interactions (Dennis and Webster 1971; Kapri and Tewari 2010; Vespermann et al. 2007). These reports are commensurate with result of the present investigation, which suggests that the production of secondary metabolites by both Trichoderma and Pseudomonas isolates have definite influence on the high degree of inhibition of P. infestans.
The mycoparasitic potential of Trichoderma spp. against P. infestans is well documented in previous findings (Ezziyyani et al. 2007; Pugeg and Ian 2006; Zegeye et al. 2011). However, in the present investigation, working with the native isolates, the mycoparasitic potential was further manifested by characteristic envelopment and coiling around the hyphae by all isolates of Trichoderma spp. The hyphae of Trichoderma spp. was also observed to grow in close proximity to the hyphae of P. infestans before coagulation and disintegration occurred.
Dixit et al. (2015) evaluated 11 isolates of fluorescent Pseudomonas for ammonia production. All isolates showed positive result for ammonia production. Lalngaihawmi and Bhattacharyya (2019) also evaluated Trichoderma spp. for ammonia production and results revealed that all the Trichoderma spp. showed positive result. These reports are in agreement with the result of the present investigation, which suggests that the production of ammonia by both Trichoderma and Pseudomonas isolates have positive impact on the plant growth.
Dixit et al. (2015) further evaluated 20 isolates of Trichoderma for IAA production. All Trichoderma spp. isolates elucidated positive results for IAA production. Prasad et al. (2017) also evaluated 24 isolates of Trichoderma spp. and 12 isolates of B. subtilis and P. fluorescens for IAA production. In the present investigation, 3 isolate of Pseudomonas and 12 isolates of Trichoderma were observed to produce IAA at varying intensity. This occurrence may be ascribed to the heterogeneous nature of the source and the strains of the antagonists.
Corbett (1974) described that HCN inhibits proper functioning of enzymes and natural receptors reversible mechanism of inhibition in the pathogens. This report is in agreement with the result of the present investigation, which suggests that the production of HCN by Pseudomonas isolates have absolute influence on the high degree of inhibition of P. infestans.
It has been observed by many investigators (Bhakthavatchalu et al. 2013; Gangwar et al. 2012; Kapri and Tewari 2010; Prasad et al. 2017; Rai 2017) that a high proportion of phosphate solubilizing microorganisms (PSMs) reside in the rhizosphere of plants and play an important role in solubilization of bound phosphates, making them available to the plants. This report is in agreement with the result of the present investigation, which suggests that the phosphate solubilization by both Trichoderma and Pseudomonas isolates have obvious influence on the plant growth.
In this present study, the strong and positive siderophore production exhibited by Pseudomonas isolates (Pf-3 and Pf-8) and Trichoderma isolates (T-3, T-4, T-5, T-7, T-8, T-9, T-10, T-11, T-14, T-15, T-18 and T-21) explicate the corresponding inhibited radial growth and high per cent inhibition of P. infestans.
Microbial consortia are known to enhance plant growth, which can result in development of various plant parts and higher growth leads to significant enhancement of vegetative growth attributes through plant growth promotion, whereas growth promotion was absent in chemical control in addition to disease suppression. Presence of consortia in the rhizoshphere increases the availability of nutrients through solubilization of insoluble sparingly soluble minerals have better nutrient uptake thereby enhancing plant growth (Biam and Majumder 2019; Harish et al. 2008; Idris et al. 2007; Raupach and Kloepper 1998; Yan et al. 2002).
Based on activities of biological control mechanism and plant growth promotion studies, the best microbial consortium was identified as MC-1. This promising indigenous liquid consortium has promoted the tomato plant growth and reduced the losses due to late blight disease in an eco-friendly manner exhibiting tremendous potential for its commercial exploitation.
References
Agrios GN (1997) Plant pathology, 4th edn. Academic Press, New York, pp 248–278
Arora RK, Sharma S, Singh BP (2014) Late blight disease of potato and its management. Potato J 41:16–40
Babu N (2006) Performance of tomato (Lycopersicon esculentum L.) as influenced by transplanting times and cultivation methods in foot hills of Nagaland. Annu Agric Res New Ser 27:159–161
Bhakthavatchalu S, Shivakumar S, Sullia SB (2013) Characterization of multiple plant growth promotion traits of Pseudomonas aeruginosa FP6, a potential stress tolerant biocontrol agent. Ann Biol Res 4:214–223
Biam M, Majumder D (2019) Biocontrol efficacy of Trichoderma isolates against tomato damping-off caused by Pythium spp. and Rhizoctonia solani (Kuhn.). Int J Chem Stud 7:81–89
Cappuccino JC, Sherman N (1992) Microbiology a laboratory manual, 3rd edn. Benjamin/Cumming Pub. Co., New York
Chowdappa P, Kumar NBJ, Madhura S, Myers KL, Fry WE, Cooke DL (2015) Severe outbreaks of late blight on potato and tomato in south India caused by recent changes in the Phytophthora infestans population. Plant Pathol 64:191–199
Corbett JR (1974) Pesticide design. In: The biochemical mode of action of pesticides. Academic Press, Inc., London, pp 44–86
Dennis C, Webster J (1971) Antagonistic properties of species groups of Trichoderma II. Production of volatile antibiotics. Trans Br Mycol Soc 57:41–48
Dixit R, Singh RB, Singh HB (2015) Screening of antagonistic potential and plant growth promotion activities of Trichoderma spp. and fluorescent Pseudomonas spp. isolates against Sclerotinia sclerotiorum causing stem rot of French bean. Legum Res 38(3):375–381
Dubos B (1987) Fungal antagonism in aerial agrobiocenoses. In: Chet I, John W, Sons (eds) Innovative approaches to plant disease control. New York, pp. 107–135
Elad Y, Chet I (1983) Improved selective media for isolation of Trichoderma spp. or Fusarium spp. Phytoparasitica 11:55–58
Ezziyyani M, Requena ME, Egea-Gilabert C, Candela ME (2007) Biological control of Phytophthora root rot of pepper using Trichoderma harzianum and Streptomyces rochei in combination. J Phytopathol 155:342–349
Gangwar M, Rani S, Sharma N (2012) Investigating endophytic actinomycetes diversity from Rice for plant growth promoting and antifungal activity. Int J Adv Life Sci 12(1):10–21
Georgakopoulos DG, Fiddaman P, Leifert C, Malathrakis NE (2002) Biological control of cucumber and sugar beet damping-off caused by Pythium ultimum with bacterial and fungal antagonists. J Appl Microbiol 92:1078–1084
Gordon SA, Weber RP (1951) Colorimetric estimation of IAA. Plant Physiol 26:192–195
Harish S, Kavino M, Kumar N, Saravanakumar D, Soorianathasundaram K, Samiyappan R (2008) Biohardening with plant growth promoting Rhizosphere and Endophytic bacteria induces systemic resistance against Banana bunchy top virus. Appl Soil Ecol 39(2):187–200
Hollomon DW (1965) Isolation of Phytophthora infestans from blighted leaves. Plant Pathol 14:34–35
Idris EES, Iglesias DJ, Talon M, Borriss R (2007) Tryptophan dependent production of indole-3-actic acid (IAA) affects level of plant growth-promotion by Bacillus amyloliquefaciens FZB42. Mol Plant Microbe Interact 20:619–626
Irzhansky I, Cohen Y (2006) Inheritance of resistance against Phytophthora infestans in Lycopersicon pimpinellifolium L3707. Euphytica 149:309–316
Kabir L, Sang WK, Yun SK, Youn SL (2013) Biocontrol of late blight and plant growth promotion in tomato using rhizobacterial isolates. J Microbiol Biotechnol 23(7):897–904
Kapri A, Tewari L (2010) Phosphate solubilization potential and phosphatase activity of rhizospheric Trichoderma spp. Braz J Microbiol 5(2):380–389
King EO, Ward MK, Raney DE (1954) Two simple media for the demonstration of pyocyanin and fluorescin. J Lab Clin Med 44(2):301–307
Kumar MSP, Chowdappa P, Krishna V (2015) Development of seed coating formulation using consortium of Bacillus subtilis OTPB1 and Trichoderma harzianum OTPB3 for plant growth promotion and induction of systemic resistance in field and horticultural crops. Indian Phytopathol 68(1):25–31
Lalngaihawmi, Bhattacharyya A (2019) Study on the different modes of action of potential Trichoderma spp. from banana rhizosphere against Fusarium oxysporum f.sp. cubense. Int J Curr Microbiol Appl Sci 8(1):1028–1040
Lamsal K, Kim SW, Kim YS, Lee YS (2013) Biocontrol of late blight and plant growth promotion in Tomato using rhizobacterial isolates. J Microbiol Biotechnol 23(7):897–904
Latijnhouwers M, Ligterink W, Vleeshouwers VG, Van West P, Govers F (2004) A Gα subunit controls zoospore mobility and virulence in the potato late blight pathogen Phytophthora infestans. Mol Microbiol 51:925–936
Loliam B, Morinaga T, Chaiyanan S (2012) Biocontrol of Phytophthora infestans, fungal pathogen of seedling damping-off disease of economic plant nursery. J Entomol. https://doi.org/10.1155/2019/324317
Milagres AM, Machuca A, Napoleao D (1999) Detection of siderophore production from several fungi and bacteria by a modification of chrome azurol S (CAS) agar plate assay. J Microbiol Methods 37:1–6
Miller RL, Higgins VJ (1970) Association of cyanide with infection of birds foot trefoil by Stemphylium loti. Phytopathology 60:269–271
Mulya K, Wataneabe M, Goto M, Takikawa Y, Tsuyumu S (1996) Suppression of bacterial wilt disease of tomato by root dipping with P. fluorescens. Annu Phytopathol Soc Jpn 62:134–140
Nautiyal CS (1999) An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiol Lett 170:265–270
Patel SS, Mukadam DS (2011) Management of plant pathogenic fungi by using Trichoderma. Biosci Discov 2(1):36–37
Prasad MR, Sagar BV, Devi GU, Triveni S, Rao SRK, Chari KD (2017) Isolation and screening of bacterial and fungal isolates for plant growth promoting properties from tomato (Lycopersicon esculentum Mill.). Int J Curr Microbiol Appl Sci 6(8):753–761
Pugeg INA, Ian DG (2006) Mycoparasitic and antagonistic inhibition on Phytophthora cinnamomi by microbial agents isolated from manure composts. Plant Pathol 5:291–298
Rai S (2017) Genomic diversity of antagonistic Trichoderma species against Phytopathogen of Lycopersicon esculentum Mill. Ph.D. Thesis, Sam Higginbottom University of Agriculture, Technology and Sciences, Allahabad, UP, India
Raupach GS, Kloepper JW (1998) Mixtures of plant growth promoting rhizobacteria enhance biological control of multiple cucumber pathogens. Phytopathology 88:1158–1164
Rodrigues J (2010) Trichoderma spp. associated with NPK fertilizer levels in pathosystem Sclerotinia sclerotiorum. M.Sc. Thesis, Federal University of Santa Maria, Santa Maria
Siddiqui IA, Shaukat SS (2003) Combination of Pseudomonas aeruginosa and Pochonia chlamydosporia for control of root-infecting fungi in tomato. J Phytopathol 151:215–222
Singh R, Ao NT, Kangjam V, Chanu NB, Daiho L, Banik S (2019) Performance assessment of native tomato genotypes to late blight disease under natural epiphytotic. Int J Curr Microbiol Appl Sci 8(11):1923–1931
Singh R, Ao NT, Kangjam V, Banik S, Sharma SK, Rajesha G, Hajong M, Lalhruaitluangi C, Daiho L (2020) Molecular characterization of potent biocontrol isolates of Trichoderma asperellum and Pseudomonas fluorescens from tomato rhizosphere. Int J Curr Microbiol Appl Sci 9(1):160–168
Sivakumar D, Wijeratnam WRS, Wijesundera RLC, Marikar FMT, Abeyesekere M (2000) Antagonistic effect of Trichoderma harzianum on post-harvest pathogens of Rambutan (Nephelium lappaceum). Phytoparasitica 28:240–247
Son SW, Kim HY, Choi GJ, Lim HK, Jang KS, Lee SO, Sung ND, Kim JC (2008) Bikaverin and fusaric acid from Fusarium oxysporum show antioomycete activity against P. infestans. J Appl Microbiol 104:692–698
Srinivasan K, Mathivanan N (2009) Biological control of sunflower necrosis virus disease with powder and liquid formulations of plant growth promoting microbial consortia under field conditions. Biol Control 51:395–402
Srinivasan K, Mathivanan N (2011) Plant growth promoting microbial consortia mediated classical biocontrol of sunflower necrosis virus disease. J Biopestic 4:65–72
Srinivasan K, Krishanraj M, Mathivanan N (2009) Plant growth promotion and the control of sunflower necrosis virus disease by the application of biocontrol agents in sunflower. Arch Phytopathol Plant Prot 42:160–172
Vespermann A, Kai M, Piechulla B (2007) Rhizobacterial volatiles affect the growth of fungi and Arabidopsis thaliana. Appl Environ Microbiol 73:5639–5641
Vincent JM (1927) Distortion of fungal hyphae in the presence of certain inhibitors. Nature 159:850
Waksman SM (1927) Principles of soil microbiology. Williams and Wikins Co., Baltimore, p 897
Waterhouse GM (1963) Key to the species of Phytophthora de Bary. Mycol Pap 92:22
Wheeler BJ (1969) An introduction to plant diseases. Wiley, New York, p 510
Wu Y, Jiang J, Gui C (2012) Low genetic diversity of Phytophthora infestans population in potato in north China. Afr J Biotechnol 11(90):15636–15642
Yan Z, Reddy MS, Ryu CM, McInroy JA, Wilson M, Kloepper JW (2002) Induced systemic resistance against tomato late blight elicited by plant growth promoting rhizobacteria. Phytopathology 92:1329–1333
Zegeye ED, Santhanam A, Gorfu D, Tessera M, Kassa B (2011) Bio-control activity of Trichoderma viride and Pseudomonas fluorescens against Phytophthora infestans under greenhouse conditions. J Agric Technol 7(6):1589–1602
Zentmyer GA (1983) The world of Phytophthora. In: Erwin DC, Bartnicki-Garcia S, Tsao PH (eds) Phytophthora: its biology, taxonomy, ecology and pathology. APS Press, The American Phytopathological Society, St. Paul, pp 1–7
Author information
Authors and Affiliations
Corresponding author
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Singh, R., Ao, N.T., Kangjam, V. et al. Plant growth promoting microbial consortia against late blight disease of tomato under natural epiphytotic conditions. Indian Phytopathology 75, 527–539 (2022). https://doi.org/10.1007/s42360-022-00464-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s42360-022-00464-1