Research Article

Evaluation of Pseudomonas fluorescens strains, Fungicides and Non-conventional Chemicals Against Botyrotinia ricini Causing Grey Mold Disease in Castor

S. Ameer Basha, V. Ramya, A. Sajeli Begum, G. Raghavendra, A. V. Ramanjaneyulu and M. V. Nagesh Kumar

  • Page No:  339 - 347
  • Published online: 07 Sep 2021
  • DOI : HTTPS://DOI.ORG/10.23910/1.2021.2293

  • Abstract

A study was made to evaluate the efficacy of Pseudomonas fluorescens strains, fungicides and non-conventional chemicals against Botryotinia ricini, causing grey mold disease in castor, under in vitro conditions. Among the 40 strains isolated from rhizosphere soil samples of different crops across the State of Telangana, India, only eight strains inhibited the growth of B. ricini under dual culture technique, of which strains Pf 21 (90.56%), Pf  23 (88.89%), Pf  34 (86.11%) and Pf 36 (84.17%) were the most effective. Among the seven chemicals (four fungicides and three non-conventional chemicals) tested for their efficacy, carbendazim followed by propiconazole had significant antagonistic effect against B. ricini. Exposure of healthy castor capsules to B. ricini and P. fluorescens for different time periods revealed that strains Pf 34 and Pf 36 were effective in completely inhibiting the growth of B. ricini and hence these two strains have been identified as effective biocontrol agents, on par with carbendazim, which offer scope for sustainable and integrated disease management of grey mold disease in castor.

Keywords :   Botrytonia ricini, biological control, castor, fungicides, Pseudomonas fluorescens

  • Introduction

    Excessive usage of chemicals like fertilizers and pesticides in agriculture has severely affected the environment and the ecosystem. Use of microorganisms as an alternative and replacement of hazardous biochemicals is an important solution for sustainable agriculture. Pseudomonas fluorescens is a widely-used plant growth promoter as well as biocontrol agent for the management of plant diseases (Panpatte et al., 2016). It  suppresses the disease through various mechanisms such as volatile antibiotics and other auxillary metabolites, siderophores, HCN while also competing with the phytopathogens for niche and nutrients through induction of systemic resistance in plants against diseases (Bhetwal et al., 2021).

    Castor is attacked by several pathogens including fungi, bacteria, viruses and nematodes throughout the world resulting in deterioration of seed quality and market value of the seeds thus causing severe losses to the farmers. Grey mold disease caused by the fungus Botryotinia ricini (Godfrey) Whetzel is a major fungal disease in castor. Though the disease is seen wherever the crop is cultivated, it is particularly troublesome for the farmers of South India, especially in Telangana, Andhra Pradesh and Tamil Nadu, where the weather conditions are more favorable for the disease development (Dange et al., 2005). The disease is responsible for disease epidemics and yield losses in Telangana state of India (Arutselvan et al., 2020).

    Spike, the economically important part in castor plant, is the primary site of grey rot infection. At later stages, infection is spread to the leaves and stem. Sussel et al. (2009) reported that if the infection occurs during flowering stage, seed filling gets affected resulting in the spike without (or less) capsule formation, thus posing great economic loss to farmers. Continuous cyclonic rainfall and high humidity are very congenial to the development of grey mold in castor (Wilcox and Seem, 1994). Sussel et al. (2011) observed a high correlation between the temperature and duration of leaf wetness with gray mold disease incidence and severity.

    Host plant resistance is an effective management strategy and researchers have been attempting to develop resistant cultivars against gray mold. So far, only cultivars with partial resistance and promising accessions (Lima and Soares, 1999; Anjani et al., 2004; Anjani and Raoof, 2010) have been identified. Cultural practices such as selection of good seed, sowing time, removal of infected plant debris and weed hosts, adoption of optimum spacing have been reported to manage the disease (Prasad, 2016; Gahukar, 2018; Soares and Cumagun, 2012, Soratto et al., 2012). Though fungicides such as propiconazole, carbendazim, azoxystrobin and iprodione were proved to be effective in controlling mold (Bezerra, 2007; Sudhakar et al., 2010; Bhat and Rajasri, 2015) the disease can be controlled only with right timing and dosage of application. 

    Biocontrol agents such as Trichoderma viride and T. harzianum were effective in inhibiting the mycelia growth of B. ricini (Bhattiprolu, 2008). Raoof et al. (2003) reported that T. viride (106 spores ml-1) application reduced the disease by 45% in detached spikes. Navaneetha et al. (2015) reported disease reduction of 65% on diseased spikes upon treatment with T. harzianum and T. asperillum. Simionato et al. (2017) and Gao et al. (2018) demonstrated the potential of Pseudomonas species in suppressing B. cinerea causing gray mold in strawberries, tomato and grapes Indira et al. (2004) reported T. viride, T. harzianum, T. hamatum, T. koeningii and P. fluorescens as potentially active biocontrol agents against gray mold disease in sorghum. Nevertheless, chemicals are also useful for farmers for immediate disease management. In castor, there is a requirement for newer or alternate disease controlling agents which can act faster and impart efficacy under field conditions. In view of this, an attempt was made to identify biocontrol agent, fungicides and non-conventional chemicals that are effective against B. ricini under laboratory conditions.

  • Materials and Methods

    2.1.  Isolation of the grey mold pathogen B. ricini

    The pathogen was isolated on Potato Dextrose Agar (PDA) medium from grey mold infected capsules of castor plants collected from Regional Agricultural Research Station, Palem, Nagarkurnool in Telangana State, India during 2015–16 and maintained at 25°C for three to four days. The culture was further purified by single spore isolation and pure culture of the fungus was maintained on PDA slants and stored at 5°C for further use (Prasad and Bhuvaneshwari, 2014).

    2.2.  Isolation of P. fluorescens from rhizosphere soil

    Soil samples from the rhizosphere of different crops such as castor, sorghum, groundnut, redgram and cotton were collected during 2015–16 from different districts of Telangana State, India.  Thirty-day old plants were selected and uprooted and the roots with enclosed soil were placed in plastic bags and stored at 4°C. For the isolation of bacteria from the rhizospheric soil, 10 g of soil was mixed with 90 ml of distilled and sterilized water. Then serial dilution of the suspension (10-1, 10-2, 10-3, 10-4, 10-5 and 10-6) was made. Around 0.1 ml from each dilution was grown on Kings B media (peptone 20 g, dipotassium hydrogen phosphate 2.5 g, glycerol 15 ml, magnesium heptahydrate 6 g, agar-agar 15 g, distilled water 1 l) and incubated at 28°C until the bacterial colonies were formed. The plates were then viewed under U.V transilluminator and fluorescent Pseudomonas colonies were identified and maintained for further use (Lamichhane and Varvaro, 2013). 

    2.3.In vitro antagonistic activity of P. fluorescens against B. ricini

    P. fluorescens strains were tested against B. ricini grown in Petri plates containing PDA medium by dual culture plate technique (Islam et al., 2018). P. fluorescens strains were streaked at a distance of 5 cm from the agar discs (5 mm in diameter) of B. ricini placed in the middle of the plate. The culture plates were sealed with plastic wrap and later the plates were incubated for eight days at 28°C. A control plate of B. ricini, without bacterial isolates was maintained and fungal growth was measured daily during the eight days of study. Zone of inhibition was measured (mm) from the edge of the mycelium to the margin of the each bacterial colony after 5 days of incubation.

    2.4.Characterization of P. fluorescens strains for antagonistic traits

    Preliminary identification of the isolates was based on colony formation, colour, pigmentation, shape and Gram staining. The strains were also characterized for production of ammonia, indole acetic acid and siderophores.

    2.4.1.  Production of ammonia

    Bacterial isolates were tested for the production of ammonia in peptone water. Freshly grown cultures were inoculated in 10 ml peptone water in each tube and incubated for 48–72 h at 36±2°C. Nessler’s reagent (0.5 ml) was added in each tube. Development of brown to yellow colour was a positive test for ammonia production (Trivedi et al., 2008).

    2.4.2.  Production of indole acetic acid

    Indole acetic acid (IAA) production was detected as described by Brick et al. (1991). Bacterial cultures were grown for 48 h on their respective media at 36±2°C. Fully grown cultures were centrifuged at 3000  rpm for 30 min. The supernatant (2 ml) was mixed with two drops of concentrated orthophosphoric acid and 4 ml of the Salkowski reagent (50 ml, 35% of perchloric acid, 1 ml 0.5 M FeCl3 solution). Development of pink colour indicated IAA production.

    2.4.3.  Siderophore production

    Siderophore production was carried out in Kings B medium using blue agar plates containing chromazurol S dye (Schwyn and Neilands, 1987). Orange color halos around the colonies on blue were indicative for siderophore production.

    2.5.  In vitro efficacy of fungicides and chemicals against B. ricini

    Four fungicides (carbendazim, mancozeb, carbendazim+ mancozeb and propioconazole) and three chemicals (oxalic acid, salicylic acid and zinc sulphate) were evaluated for their efficacy on mycelial growth of B. ricini by poisoned food technique (Schmitz, 1930). Each fungicide and chemical was dispensed separately in molten sterilized PDA medium at seven concentrations (300, 500, 700, 900, 1000, 2000 and 3000 ppm), obtained by adding sterile water to the stock solutions. Five mm mycelia disc, taken from one week old culture of the B. ricini, were asceptically placed in the center of solidified poisoned PDA. Five replications were maintained for each concentration. The Petriplates were incubated at 24°C and observations of the mycelial growth of B. ricini were recorded after seven days of incubation. The growth of the fungus on non-poisoned PDA served as a control. The % inhibition in the growth due to various fungal treatments at various concentrations was computed as follows:

    Mycelial growth inhibition (%)=[(dc-dt/dc)] x 100 (%).

    where, dc=Average diameter of fungal colony in control, and

    dt=Average diameter of fungal colony in treatment group

    2.6.  Efficacy of P. fluorescens against B. ricini using detached capsule technique

    Based on in vitro experimental data, five strains of P. fluorescens that were effective in inhibiting the mycelial growth of B. ricini along with the most effective fungicide (or chemical), were tested for their efficacy against B.ricini using detached capsule technique (Prasad et al., 2016). Fruit capsules of 15–20 days old were detached from healthy castor spikes, surface sterilized and dipped in a suspension of B. ricini and bioagent/fungicide for 0, 1, 4, 6 and 12 hours. For each treatment, four capsules were maintained at 20–25°C and 90% relative humidity in petri plate for a week in three replications. Capsules treated with B. ricini were positive control and uninoculated capsules constituted negative control. The number of capsules infected was recorded each day after the appearance of first symptom.

  • Results and Discussion

    3.1.  Isolation of the grey mold pathogen B. ricini

    B. ricini was isolated from grey mold infected castor plants collected from Regional Agricultural Research Station, Palem, Nagarkurnool in Telangana State (Figure 1). The fungus produced pluffy mycelial growth on PDA without sporulation. This is in partial agreement with the observations made by Prasad et al. (2016) who also reported pluffy mycelia growth but with enhanced sporulation of B. ricini on oat meal agar medium.

    3.2.  Isolation of P. fluorescens from rhizosphere soil

    Forty strains of P. fluorescens were isolated from the rhizosphere soil samples of  castor, cotton, groundnut, rice and redgram crops in Nagarkurnool district, Telangana State. This is similar to the results reported in rice where thirty P. fluorescens isolates were collected from rhizosphere soils (Reddy et al.,2007). The present reporting of 40 P. fluorescens assumes significance because there are limited reports on evaluation of so many different isolates of P. fluorescens against B. ricini. Though there are several studies on use of other biological control agents like Trichoderma spp. to control gray mold in castor (Bhattiprolu and Bhattiprolu, 2006; Chagas, 2009; Raoof et al., 2003; Tirupathi et al., 2006), there are limited studies on use of P. fluorescens. Raoof et al. (2003) reported the use of both P. fluoresens and T. viride to manage B. ricini, however, single isolates were evaluated in the study compared to the forty isolates used in this study.Navaneetha et al. (2015) reported the use of B. ricini using T. harzianum and T. asperellum isolates. Bhattiprolu and Bhattiprolu (2006) evaluated the effectiveness of a single isolate of T. viride against B. ricini as influenced by culture medium pH, incubation temperature, fungicides and exposure to mutagenic agents (gamma rays). 

    3.3.  In vitro antagonistic activity of P. fluorescens against B. ricini

    Among the 40 strains of P. fluorescens tested for their antagonistic activity against B. ricini, only eight strains (Pf 3, Pf 4, Pf 21, Pf 22, Pf 23, Pf 24, Pf 34 and Pf 36) showed zone of inhibition against B. ricini. Maximum inhibition (90.56%) was recorded by Pf 21 followed by Pf 23 (88.89%).  These two strains were isolated from rhizosphere soils of rice and groundnut respectively. Strains Pf 34 and Pf 36 recorded 86.11% and 84.17% inhibition respectively. Minimum inhibition was observed with Pf 3, Pf 4, Pf 22 and Pf 24 (Figure 2).

    Interestingly Pf3 and Pf4 were isolated from the rhizosphere soils of castor crop (Table 1). P.fluorescens strains were proved to be effective in inhibiting the growth of B. ricini. The results of this study were in conjunction with the earlier reports on the effect of P.fluorescens against B.ricini in castor (Singh, 2014). In castor, in addition to P. fluorescens, the importance of Trichoderma spp. in integrated disease management against grey mold weas reported by Navaneetha et al. (2015). They demonstrated the effectiveness of T. harzianum and T. asperellum isolates against B. ricini  by 55–65%. Similarly, Bhattiprolu and Bhattiprolu (2006) showed 60% inhibition of B. ricini using poisoned food technique. A gamma ray mutant isolate of T. viride TV4 obtained the highest inhibition of B. ricini of about 88% when compared to the wild isolate (Bhattiprolu, 2008). In rice, Reddy et al.(2007) reported one isolate (Pf 003) of P. fluorescens among the 30 isolates which effectively inhibited the growth of four rice pathogens viz., Magnaporthe grisea, Drechslera oryzae, Rhizoctonia solani and Sarocladium oryzae by 62 to 85%.

    3.4.  Characterization of P. fluorescens strains for plant growth promoting traits

    All the eight strains i.e., Pf 3, Pf 4, Pf 21, Pf 22, Pf 23, Pf 24, Pf 34 and Pf 36) took about 24 hours to establish their growth on kings B agar medium. Colonies were dull white in color, small to medium in size, smooth and glistening with yellowish green to light green pigmentation. The strains were Gram negative, small, single isolated rods without sporulation. The strains were tested for antagonisitic properties viz., HCN, ammonia, IAA and siderophore production. Strains Pf 21 and Pf 34 tested positive for HCN production (Table 1; Figure 3a).

    Similar results were reported by Singh et al. (2015), where in change in color after incubation from yellow to light brown indicated weak HCN production and change in color from yellow to reddish brown indicated strong HCN production. They reported that isolates PSF 1, PSF 3, PSF 8 and PSF 12 produced highest HCN. Bano and Mussarat (2003) reported efficient HCN producing Pseudomonas aeruginosa strain NJ-15 as a potential biocontrol agent. Bhoonobtong et al. (2012) also reported the anti-microbial activity of P. aeruginosa isolated from medicinal plants in Thailand. All the eight isolates tested positive for ammonia production (Table 1). Four isolates viz., Pf 3, Pf 4, Pf 22 and Pf 24 tested positive for IAA (Figure 3b) and four isolates viz., Pf 21, Pf 23, Pf 34 and Pf 36 tested positive for siderophore production (Figure 4). Similarly, fluorescent Pseudomonas strains were isolated from the rhizosphere soils of various crops namely castor, redgram, rice, greengram, sunflower, potato etc. and characterized for their morphological, cultural and biochemical attributes (Gupta et al., 2002; Bhatia et al., 2008).

    3.5.  In vitro efficacy of fungicides and chemicals against B. ricini

    Four fungicides tested effectively inhibited mycelial growth of B.ricini. There was a significant decrease in mycelial growth with increase in the concentration of fungicide (Table 2). Carbendazim completely inhibited (100%) the growth of B. ricini at all the tested concentrations including low concentrations of 300, 500, 700, 900 and 1000 ppm compared to the recommended dose of 1 g l-1 (Yasmeen et al., 2003) Interestingly, the combination fungicide viz., carbendazim+mancozeb recorded complete inhibition at concentration 700 ppm and above which is much lower than the recommended dosage of 1.5 g l-1. Propioconazole was also toxic to B. ricini with a significant inhibitory effect at all concentrations and showed complete inhibition of B. ricini at concentrations 2000 and 3000 ppm. All the three tested chemicals showed significant inhibition of B. ricini, with oxalic acid showing complete inhibition at 700 ppm and above. Salicylic acid and ZnSo4 recorded no growth at 2000 and 3000 ppm concentration with negligible growth observed up to 1000 ppm concentration (Table 2).

    Niki (2003) observed that carbendazim was effective in inhibiting Botrytis sp. under glass house conditions. Simiarly, Bezerra (2007) also reported that carbendazim was effective against grey rot under controlled conditions. Chagas (2009) reported that carbendazim, tebuconazole, iprodione and procymidone were highly effective against grey rot under in vitro and in vivo conditions. Fungicides such as armure, thiram, edifenphos and carbendizam and their mixtures were proved effective against B. ricini mycelia growth rate and spore germination under in vitro conditions. Haenssler et al. (2000) observed that fenhexamid was effective against grey rot under green house conditions. In addition to fenhaxamid, newer fungicides namely pyrimethanil, fludioxonil, fluazinam etc. are also being used along with carbendizam (Couderchet, 2003).

    3.6.  Efficacy of P. fluorescens and fungicides against B. ricini using detached capsule technique

    Symptoms of B. ricini infection characterized by grayish cottony mycelial growth appeared on castor capsules on 3 days after inoculation (DAI). By 6 DAI, the infected capsules were fully covered by the fungus. In 0 hour treated set, there was no infection of B. ricini. Similarly, in uninoculated control (without B. ricini) and only P. fluorescens (without B. ricini) treated samples, the capsules appeared healthy without any pathogen infection. Overall, increased exposure time to P. fluorescens and carbendazim reduced B. ricini infection. Capsules treated with carbendazim for 1 hour recorded 25% infection by 6 DAI compared to zero infection in capsules exposed for 4, 6 and 12 hours (Figure 5). Within a given exposure time, P. fluorescens strains differed in their efficacy to inhibit B. ricini infection on castor capsules. In castor capsules exposed for 1 hour to P. fluorescens and B. ricini, strain Pf 36 recorded 50% infection by 6 DAI compared to 100% infection for Pf 22. In capsules exposed to P. fluroescens and B. ricini for 6 hours, Pf 21, Pf 34 and Pf 36 recorded 50% capsule infection compared to 75% infection for Pf 23 and Pf 22. Both Pf 34 and Pf 36 were effective in completely controlling B. ricini growth on castor capsules when exposed for 12 hours followed by strain Pf 21 with 25% infection. Strain Pf 22 was least effective with 75% capsules infected with B. ricini (Figure 5). Raoof et al. (2003) reported that Trichoderma spp. and P. fluorescens controlled castor gray mold to an extent of 40–50% under artificial inoculation conditions.

    The evaluation of antifungal activity of different fungicides in vitro condition is important for integrated disease management where limited and appropriate quantity of agrochemical should be used for achieving eco friendly management of grey rot. The effectiveness of P. fluorescens strains in inhibiting the growth of B. ricini has been clearly demonstrated with Pf 34 and Pf 36 being the most effective strains. Both the isolates tested positive for siderophore production. P. fluorescens secretes a hydroxamate-type siderophore which was effective in controlling Macrophomina phaseolina in groundnut (Gupta et al., 2002) through their ability to chelate and reduce the amount of ferric ions available in rhizosphere, competing with the plant pathogens and thus reduce their ability to colonize plant roots. P. fluorescens induces plant systemic resistance, which reduces the pathogen infection (Abozaid et al., 2020). The siderophore producing Pf 34 and Pf 36 strains were most effective in inhibiting B. ricini on castor capsules at 12 hours exposure period.

  • Conclusion

    Eight strains out of all the tested P. fluorescens were antagonistic to castor grey mold pathogen B. ricini. Among the chemicals, carbendazim showed maximum efficacy on the growth of B. ricini and in lowering capsule infection. The effectiveness of P. fluorescens strains Pf 34 and Pf 36 in inhibiting the growth of B. ricini on castor capsules at 12 hours exposure time has been clearly demonstrated and was on par with fungicide treated.

  • Further Research

    Since the present study was conducted under laboratory conditions, more insight is required into the effect of exposure time of B. ricini spayed with bacterial (potential strains P. fluorescens) suspension alone or in combination with fungicides and non- conventional chemicals on disease infection under field conditions, more so because grey mold is a weather dependent disease. This would immensely help in taking prophylactic measures at appropriate time before the onset of the cyclonic rainfall and subsequently reduce the disease infection.

  • Reference
  • Abozaid, G.A., Soliman, N.A., Abdullah, A.S., El-Sharouny, E.E., Matar, S.M., Sabry, S.A.F., 2020. Maximization of siderophores production from biocontrol agents, Pseudomonas aeruginosa F2 and Pseudomonas fluorescens JY3 using batch and exponential fed-batch fermentation. Processes 8, 455–475.

    Adefris, T., Nigussie A., 1993. Castor in Ethiopia: production, utilization and research. Oilcrops Newsletter 10, 40–43.

    Anjani, K., Raoof, M.A., Ashoka, P., Reddy, V., Hanumanta Rao, C., 2004. Sources of resistance to major castor (Ricinus communis) diseases. Plant Genetic Resources Newsletter 137, 46–48.

    Anjani, K., Raoof, M., 2010. Sources of resistance to Botrytis ricini (Godfrey) in castor germplasm. Journal of Oilseeds Research 27(1), 20–21.

    Arutselvan, R., Prasad, R.D., Uma Devi, G., Sarada, C., 2020. Effect of temperature and culture media on mycelia growth and sporulation of Botryotinia ricini causing grey mold of castor. Journal of Research, PJTSAU, 48(1&2), 21–27.

    INDIASTAT, 2019. Department of agriculture and cooperation, Govt of India. Availabe from Accessed 27 October 2020.

    Bano, N., Musarrat, J., 2003. Characterization of a new Pseudomonas aeruginosa strain NJ-15 as a potential biocontrol agent. Current Microbiology 46(5), 324–328.

    Bezerra, C.S., 2007. Estrutura genetica e sensibilidade a fungicidas de Amphobotrys ricini agente causal do mofo cinzento da mamoneira. MSc dissertation (Genetic and molecular biology), Universidade Federal do Rio Grande do Norte, Natal, Brazil.

    Bhat, B.N., Rajasri, M., 2015. Management of botrytis grey mold of castor with chemicals and biocontrol agents. Progressive Research 10(2), 129–131.

    Bhatia, S., Maheswari, D.K., Dubey, R.C., Arora, S., Bajpai, V.K., Kang, S.C., 2008. Beneficial effects of fluorescent Pseudomonads on seed germination, growth promotion and suppression of charcoal rot in groundnut (Arachis hypogea L.). Journal of Microbiology and Biotechnology 18(9), 1578–1583.

    Bhattiprolu, S., 2008. Growth of Trichoderma viride as influenced by pH, temperature, botanicals, fungicides and mutagenic agent. Indian Journal of Plant Protection 36(2), 279–282.

    Bhattiprolu, S.L., Bhattiprolu, G.R., 2006. Management of castor gray rot disease using botanical and biological agents. Indian Journal of Plant Protection 34(1), 101–104.

    Bhetwal, S., Rijal, R., Das, S., Sharma, A., Pooja, A., Malannavar, A., 2021. Pseudomonas fluorescens: Biological control aid for managing various plant diseases: A Review. Biological Forum 13(1), 484–494.

    Bhoonobtong, A., Sawadsitang, S., Sodngam, S., Mongkolthanaruk, W., 2012. Characterization of endophytic bacteria, Bacillus amyloliquefaciens for antimicrobial agents production. International Conference on Biological and Life Sciences 40, 6–11.

    Brick, J.M., Bostock, R.M., Silverstones, S.E., 1991. Rapid in situ assay for indole acetic acid production by bacteria immobilized on nitrocellulose membrane. Applied and Environmental Microbiology 57(2), 535–538.

    Chagas, H.A., 2009. Controle de mofo-cinzento (Amphobotrys ricini) da mamoneira (Ricinus communis L.) por metodos quimico, biologico e com oleos essenciais. MSc Dissertation, (Agronomy), Universidade Estadual Paulista “Júlio de Mesquita Filho”, Botucatu, Brazil.

    Couderchet, M., 2003. Benefits and problems of fungicide control of Botrytis cinereain vineyards of Champagne. Vitis42(4), 165–171.

    Damodaran, T., Hegde, D.M., 2002. Oilseeds situation: a statistical compendium. Directorate of Oilseeds Research, Hyderabad, 210–229.

    Dange, S.R.S., Desai, A.G., Patel., S.I., 2005. Diseases of castor. In: Saharan, G.S., Mehta, N., Sangwan, M.S. (Eds), Diseases of oilseed crops. Indus Publishing Co, New Delhi India, 211–234.

    Gao, P., Qin, J., Li, D., Zhou, S., 2018. Inhibitory effect and possible mechanism of a Pseudomonas strain QBA5 against gray mold on tomato leaves and fruits caused by Botrytis cinerea. PLoS ONE, 13(1), e0190932.

    Gahukar,  R.T., 2018. Management of pests and diseases of castor (Ricinus communis L.) in India: current status and future perspective. Archives of Phytopathology and Plant Protection 51(18), 956–978.

    Gobin, A.M.L., Uguru, M.I., Deckers, J., 2001. Castor. In: Reamaekers, R.H. (Ed), Crop Production in Tropical Africa, 725–733.

    Gupta, C.P., Dubey, R.C., Maheshwari, D.K., 2002. Plant growth enhancement and suppression of Macrophomina phaseolina causing charcoal rot of peanut by fluorescent pseudomonas. Biology and Fertility of Soils 35, 399–405.

    Indira, S., Venkateshwaran, M., Tonapi, V., Varanavasiappan, S., Seetharama, N., 2006. Utility of biocontrol agents in the suppression of seed borne pathogenic mycoflora and their effect on seed quality in sorghum (Sorghum bicolor (L.) Moench). Archives of Phytopathology and Plant Protection 39, 379–387. 

    Islam, M.A., Nain, Z., Alam, M.K., Banu, N.A., Islam, M.R., 2018. In vitro study of biocontrol potential of rhizospheric Pseudomonas aeruginosa against Fusarium oxysporum f. sp. cucumerinum. Egyptian Journal of Biological Pest Control 28, 90–101. 

    Haenssler, G., Study, A., Stuebler, D., 2000. Uptake, sensitivity profile and biochemical mode of action of the new Botryticide fenhexamid. In: Proceedings of XII International Botrytis Symposium. Reihms, 33.

    Lamichhane, J.R., Varvaro, L., 2013. A new medium for the detection of fluorescent pigment production by pseudomonads. Plant Pathology 62, 624–632.

    Lima, E.F., Soares, J.J., 1990. Resistance of castor bean cultivars to gray mold caused by Botrytis ricini. Brazilian Phytopathology 15(1), 96–98.

    Navaneetha, T., Prasad, R., Rao, L.V., 2015. Liquid formulation of Trichoderma species for management of gray mold in castor (Ricinus communis L.) and Alternaria leaf blight in sunflower (Helianthus annuus L.). Journal of Biofertilizers and Biopesticides 6, 1–11.

    Niki, P.P., Markellou, E., Kalamarakis, A.E., Kyriakopoulou, D., Malathrakis, N.E., 2003. In vitro and in vivo activity of cyprodinil and pyrimethanil on Botrytis cinerea isolates resistant to other botryticides and selection of resistance to pyrimethanil in a greenhouse population in Greece. European Journal of Plant Pathology 109, 173–182.

    Panpatte, D., Jhala, Y., Shelat, H., Vyas, R.V., 2016. Pseudomonas fluorescens: A Promising biocontrol agent and PGPR for sustainable agriculture. In: Singh, D.P. et al. (Eds.), Microbial inoculants in sustainable agricultural productivity, 257–270.

    Prasad, R.D., 2016. Gray mold of castor. Indian council of agricultural research-Indian Institute of Oilseeds Research, Hyderabad, 1–40.

    Prasad, R.D., Bhuvaneswari, R., 2014. A modified medium for improved sporulation of gray mold pathogen, Botryotinia ricini (Godfrey) Whetzel in castor (Ricinus communis L.) Journal of Oilseeds Research 31(1), 79–81.

    Prasad, R.D, Raoof, M.A., Senthilvel, S., Dinesh Kumar, V., Praduman, Y., Bhuvaneshwari, R., Varaprasad, K.S., 2016. Gray mold of castor. Indian Council of Agricultural Research-Indian Institute of Oilseeds Research, Rajendranagar, Hyderabad.

    Raoof, M., Yaseem, M., Kausar, R., 2003. Potential of biocontrol agents for the management of castor grey mold, Botrytis ricini godfrey. Indian Journal of Plant Protection 31(2), 124–126.

    Reddy, K.R.N., Choudary, D.A., Reddy, M., 2007. Antifungal metabolites of pseudomonas fluorescens isolated from rhizosphere of rice crop. Journal of Mycology and Plant Pathology 37, 280–284.

    Schmitz, H., 1930. Poisoned food technique. Industrial and Engineering Chemistry Analyst Ed. 2, 361.

    Schwyn, B., Neilands, J.B., 1987. Universal chemical assay for the detection and determination of siderophores. Analytical Biochemistry 160, 47–56.

    Sharma, D.K., Shafkat, R., 2017. Seed-borne and post-harvest diseases of castor bean (Ricinus communis Linn.) and their management: A review. Journal of Phytological Research 30(1), 31–45

    Simionato, A.S., Navarro, M.O.P., de Jesus, M.L.A., Barazetti, A.R., da Silva, C.S., Simoes, G.C., Balbi-Pena, M.I., de Mello, J.C.P., Panagio, L.A., de Almeida, R.S.C., Andrade, G., de Oliveira, A.G., 2017. The effect of phenazine-1-carboxylic acid on mycelial growth of Botrytis cinerea produced by Pseudomonas aeruginosa LV strain. Frontiers in Microbiology 8, 1102.

    Singh, H.B., 2014. Management of plant pathogens with microorganisms. Proceedings of National Academy of Sciences, India 80(2), 443–454.

    Singh, N., Ghosh, M., Singh, D., Raina, S., Helfish, A.I., 2015. Antagonistic activity of indigenous strains of Pseudomonas strains against fungal plant Pathogens. Journal of Pure and Applied Microbiology 9(3), 2255–2259.

    Soares, D.J., Cumagun, C., 2012. The gray mold of castor bean: a review. Plant pathology. In Tech Publisher, Rijeka, Croatia, 219–240.

    Soratto, R.P., Souza-Schlick, G.D., Fernandes, A.M., Zanotto, M.D., Crusciol, C.A., 2012. Narrow row spacing and high plant population to short height castor genotypes in two cropping seasons. Industrial Crops and Products 35(1), 244–249.

    Sudhakar, R., Khayum Ahammed, S., Meena Kumari, K., Srinivas, T., Vishnuvardhan Reddy, A., 2010. Integrated disease management of botrytis grey rot of castor (Ricinus communis L.). Journal of Oilseeds Research 27, 239–240.

    Sussel, A.A.B., Pozza, E.A., Castro, H.A., 2009. Elaboration and validation of diagrammatic scale to evaluate gray mold severity in castor. Tropical Plant Pathology 34(3), 186–191.

    Sussel, A.A.B., Pozza, E.A., Castro, H.A., Lasmar, E.B.C., 2011. Incidence and severity of castor bean gray mold based on temperatures, wetting periods and concentration of conidia. Summa Phytopathologica 37(1), 30–34.

    Tirupathi, J., Kumar, C.P.C.,  Reddy, D.R.R., 2006. Trichoderma as potential biocontrol agents for the management of grey mold of castor. Journal of Research ANGRAU 34(2), 31–36.

    Trivedi, P., Pandey, A., Palni, L.M.S., 2008. In vitro evaluation of antagonistic properties of Pseudomonas corrugata. Microbiological Research 163, 329–336.

    Wilcox, W., Seem, R., 1994. Relationship between strawberry gray mold incidence, environmental variables, and fungicide applications during different periods of the fruiting season. Phytopathology 84, 264–270.

    Yasmeen, M., Raoof, M.A., Rana, K., 2003. Host range of Botrytis ricini Godfrey, castor gray mold pathogen. In national symposium on “plant pathogens diversity in relation to plant health” held at Osmania University, Hyderabad, India).

People also read

Short Research

Preference Towards Online Mode of Distance Education Courses–conjoint Analysis

M. Malarkodi, V. M. Indumathi and S. Praveena

Conjoint analysis, distance education, preference, online learning

Published Online : 07 Feb 2018