2
Formulation Technology of Biocontrol Agents: Present Status and Future Prospects Chetan Chetan Keswani, Keswani, Kartikay Kartikay Bisen, Vivek Singh, Birinchi Kumar Sarma, and Harikesh Bahadur Singh
Contents
Abstract
2.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 2.2 2.2. 2.2.1 1 2.2.2
. . . . . . . . . .. . . . . 37 Types of Formulations . . . . . . . . . .. Wett Wettab able le Powd Powder erss and and Liqu Liquid idss .. .. .. .. .. .. .. . 37 Granular Formula mulati tio ons .. .. .. .. .. .. .. .. .. .. .. . 37
2.3
Biocontrol Products Containing Fungi and Their Formulations . . . . . . . . . . . . . . . . . . . . 39
2.4
Biocontrol Products Containing Bacteria and Their Formulation . . . . . . . . . . . . . . . . . . . . . 41
2.5
Seed Treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
2.6
Carriers Carriers and Adjuvant Adjuvant Used in Microbial Microbial Formulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
2.7
Basic Information Required for Microbial Product Registration . . . . . . . . .. . . . . . . . . .. .. . . . . . . Stra train Specifications .. .. .. .. .. .. .. .. .. .. .. .. .. Shelf Life and Storage ... .. .. ... .. .. .. ... ... .. CFU Count ... .... .... .... ... ... ... .... .... .... Path athogen ogenic ic Cont Contam amin inat atio ion n .. .. .. .. .. .. .. .. .. . Moisture Content ... ... ... ... ... ... .. ... ... ...
2.7.1 2.7.2 2.7.3 2.7. 2.7.4 4 2.7.5
44
46 46 46 46 47 47
2.8
Constraints in the Production of Microorganism-Based Biopesticides . . . . 47
2.9
. . 47 Future Prospects . . . . . . . . . . . . . . . . . . . . . . . . . . ..
2.10 Conclusion . . . . . . . . . . . .. . . . . . . . . . . . . . . . .. . . . . . . . . . 48 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
49
C. Keswani • K. Bisen • V. Singh • B.K. Sarma • H.B. Singh (*) Department of Mycology and Plant Pathology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi 221005, India e-mail:
[email protected] e-mail:
[email protected]
Recent shift in trends of agricultural practices from application of synthetic fertilizers and pestici pesticides des to organi organicc farmin farming g has brough broughtt into into focus focus the use use of microo microorga rganis nisms ms those those carryout carryout analogous analogous functions functions.. Formulatio Formulations ns of rhizomicro rhizomicroorgan organisms isms available available in global global markets markets range from talc-based talc-based and liquid and secondary secondary metabolitemetabolite-based based formulation formulations. s. The ideal conditions required for development of high efficiency formulations of biopesticides include selection of potent strains, shelf life, storage, application technology, quality control, trol, biosaf biosafety ety,, and regist registrat ration ion.. In this this chapte chapter, r, we will discuss the constraints associated with deve develo lopm pmen entt and and comm commer erci cial aliz izat atio ion n of bioino bioinocul culant ants. s. Moreo Moreover ver,, specia speciall emphas emphasis is will be on the next generation of antimicrobial secondary metabolite formulations which will not only have a much longer shelf life but also a higher efficiency against soilborne rne phytopathogens particularly against bacteria; also, a consortium of antimicrobial meta metabo boli lite tess agai agains nstt indi indivi vidu dual al path pathog ogen enss could be formulated and used regardless of geogra geographi phicc locatio location n where where the incide incidence nce of that particular disease is high. This approach would be unsurpassed by current technology, as the formulation would specifically target a part partic icula ularr path pathog ogen en whil whilee remai remaini ning ng soil soil microbiota would remain unaffected.
# Springer India 2016
N.K. Arora et al. (eds.), Bioformulations: for Sustainable Agriculture , DOI 10.1007/978-81-322-2779-3_2
35
36
C. Keswani et al.
biop biopes esti tici cide dess was was arou around nd 0.2 0.2 % of the the tota totall pesticides’ market during the year 2000, and it Convention Conventional al farming farming around around the world world is primarprimar- amplified to 4.5 % by 2010. The market value is ily based on chemical fertilizers and pesticides for expected to reach around US$ 1 billion (Singh al. 2012 2012). ). In India, India, curren currently tly 34 microorg microorgani anisms sms plant plant nutriti nutrition on and diseas diseasee manageme management, nt, a practi practice ce et al. which pounded huge negative impacts on human have been included in the schedule of Gazette of and environment health. Globally, rising aware- India for registration as biopesticide with Central Insecticid icidee Board, Board, Farida Faridabad bad,, under under section sectionss 9 ness ness of the the haza hazarrdous dous effe effect ctss of synt synthe heti ticc Insect (3B) and and 9(3) 9(3) of the the Inse Insect ctic icid ides es Act, Act, 1968 1968 pest pestic icid ides es has has incr increa ease sed d the the dema demand nd for for safe safer r (3B) (Table 2.1)) (Keswani et al. 2013a al. 2013a). ). alternatives alternatives.. Various Various microorgani microorganisms sms are currently currently (Table 2.1 Variou Variouss clas classe sess of bioc biocon ontr trol ol agent agentss have have being explored and utilized as biological control shown n sign signifi ifica cant nt anta antago goni nism sm to a rang rangee of agents agents (BCAs) (BCAs) or biopesti biopesticid cides. es. Popul Popular ar BCAs BCAs show include Trichoderma spp., Pseudomonas fluores- phytopathogens in vitro, but generally they have cence, Bacillus spp., Ampelomyces Ampelomyces quisqualis quisqualis, irregular performance in field conditions. Various factors are respon responsib sible le for inconsi inconsiste stent nt perfor perfor- Agrobacteriu Agrobacterium m radiobacter radiobacter , nonp nonpat atho hoge geni nicc factors Fusarium Fusarium, Coniothyrium, and atoxigen atoxigenic ic Aspergil Aspergil- mance which includes poor shelf life, susceptibillus niger (Singh 2006 (Singh 2006;; Keswani et al. 2014 al. 2014,, 2015; ity of microbial strain to various abiotic stresses, Mish Mishra ra et al. al. 2015 2015). ). Approx Approxima imatel tely, y, there there are 1400 1400 and low organic carbon content in the soil. Forbiop biopes esti tici cide de prod produc ucts ts bein being g sold sold worl worldw dwid idee mulation technologies are used for stabilizing the (NAAS 2013) 2013). The tot total market share of microorganisms during production, storage, and
2.1
Int Introdu oductio ction n
Microbes listed in the Gazette of India for production production of biopesticide biopesticidess and registration registration under sections sections 9 Table 2.1 Microbes (3B) and 9(3) of the Insecticides Act, 1968 Bacterial
Fungal
Viral
Burkholderia cepacia strain 84 Agrobacterium radiobacter strain Agrobacterium tumefaciens Erwinia amylovora (hairpin protein) Alcaligenes spp.
Verticillium chlamydosporium Streptomyces griseoviridis Streptomyces lydicus Candida oleophila Fusarium oxysporum (non pathogenic) Penicillium islanidicum (for groundnut) Pythium oligandrum VAM ( fungus) Trichoderma spp. – strain Aspergillus niger – strain AN27 Gliocladium spp. Beauveria bassiana Verticillium lecanii
Granulosis viruses Nuclear polyhedrosis viruses (NPV)
Photorhabdus luminescences akhurustii strain K -1 Photorhabdus luminescences Serratia marcescens GPS 5 Bacillus subtilis Pseudomonas fluorescens
Metarhizium anisopliae Nomuraea rileyi Hirsutella sp. Ampelomyces quisqualis Phlebia gigantean Coniothyrium minitans Chaetomium globosum Myrothecium verrucaria Paecilomyces lilacinus Piriformospora indica
2
Formulation Technology of Biocontrol Agents: Present Status tus and Future Prospects
dist distri ribu buti tion on,, aidi aiding ng in the the appl applic icat atio ion n and and hand handli ling ng,, prot protec ecti ting ng the the micro microor orga gani nism sm from from damaging environmental factors, and enhancing the activity of the organism (Jones and Burges 1997). 1997 ). In a micr microb obia iall form formul ulat atio ion, n, the the majo major r focus is to preserve microorganism for enhancing thei theirr anta antago goni nism sm agai agains nstt targ target et path pathog ogen ens. s. Potency Potency of microbi microbial al formul formulatio ation n is primaril primarily y dependant on the strain of microorganism used, thou though gh ther theree may may be cruc crucia iall phys physic ical al and and nutritional requirements of the microorganism to rema remain in acti active ve for for long longer er time time.. Benefi enefici cial al microorganis microorganisms ms are considered considered as eco-friendl eco-friendly, y, and it is mandatory that any additives in the formulation should be eco-friendly. Commercial success of these formulations is based on the capability of a microorganism to survi survive ve and prolif prolifera erate te in the field field condit condition ion,, shelf life, and efficiency to control pest and disease, market price, ease of handling, and application cation (Lisansky (Lisansky 1985). 1985). Decisi Decision on of select selecting ing formulation depends fundamentally on the target organism to be managed, as well as on the ecology and biology of the biocontrol agents and host plant (Jacobsen and Backman 1993 1993). ). Moreover, the best feature of this approach is that it can easily be integrated with different pest management modules. BCAs have been formulated in various ways such as wettable powders, liquid, and granules for application such as sprays, seed treatments, drenches, and dips and incorporation into soil and pot mix.
37
in wett wettab able le powd powdeer, liqui iquid, d, and granu anular formul formulatio ations ns (Singh (Singh et al. 2012 2012,, 2014 2014)) (Table 2.2 2.2). ). Dusts (DP) are formulated by adding an active ingredient on fine solid mineral powder such as clay and talc with particle size ranging from 50 to 100 μm. Dusts are applied directly to the target, either manually or mechanically. Inert ingredients used for dust formulation are anticaking agents, ultraviolet protectants, and adhesive materials to enhance adsorption. Dusts usually contain <10 % of microorganisms by weight. Granule (GR) particles are heavier and larger compared to dust. Microgranules (100–600 μm) and and coar coarse se part partic icle less size size (100 (100–1 –100 000 0 μm) are made made from from mine minera rall mate materi rial alss such such as sili silica ca,, kaol kaolin in,, star starch ch,, atta attapu pulg lgit ite, e, poly polyme mers rs,, grou ground nd plan plantt residues, and dry fertilizers (Tadros 2005 2005). ). Concentration of microorganisms in granules ranges from 5 to 20 %. Three types of granule formul formulati ations ons are curren currently tly availab available: le: (1) the micromicroorganism is sprayed on a rotating granular carrier withou withoutt a sticke sticker, r, (2) the microo microorga rganis nism m is attach attached ed to the the oute outerr surf surfac acee of a gran granul ular ar carr carrie ierr by a sticker, and (3) the microorganism is incorporated into a carrier paste or powder as a matrix.
2.2.1 2.2.1
Wettab Wettable le Powder Powderss and and Liqui Liquids ds
BCAs such BCAs such as Bacillus Bacillus subtilis subtilis, Pseudomonas putida, and Trichoderma spp. are used to control various diseases and are generally applied as dip or dren drenche chess and and spra sprays ys to frui fruitt afte afterr harv harves estt (Tronsmo and Dennis 1983 1983;; Colyer and Mount 1984;; Pusey and Wilson 1984 1984 Wilson 1984;; Wilson and Pusey 2.2 2.2 Type Typess of Form Formul ulat atio ions ns 1985). 1985 ). Spraying of Penicillium sp. to pineapple fruit result resulted ed in reduce reduced d postha postharve rvest st diseas diseases es Although Although pestic pesticide idess are formul formulated ated in various various fruit ways ways includi including ng dry formulat formulation ionss such such as dusts dusts (Lim and Rohrbach 1980 Rohrbach 1980). ). (DP), (DP), granul granules es (GR), (GR), and microgr microgranu anules les (MG); (MG); seed dressing formulations such as powders for seed dressing (DS); dry formulations for dilution 2.2.2 2.2.2 Granu Granular lar Formul Formulati ations ons in water including dispersible granules (WG) and wettable powders (WP); liquid formulations for Lignite silage was applied to produce granules Trichod oderm erma a harzi harzian anum um dilution in water such as emulsions, suspension containing Trich and concentrates (SC), oil dispersions (OD), and cap- Gliocl Gliocladi adium um roseum roseum to cont contro roll Rhizoctonia sule sule susp suspen ensi sion onss (CS) (CS);; and and ultr ultralo alow w volu volume me solani in soil soil causin causing g dampin damping-o g-off ff of peanut peanut formulation formulationss (Knowles (Knowles 2005, 2005, 2006) 2006) howe howeve ver, r, (Jones et al. 1984 al. 1984). ). Lignite was grinded to proglobally biopesticides available in the market are duce granules of 425–2000 μm in diameter and
38
C. Keswani et al.
Table 2.2 Types of pesticide formulations formulations (Modified (Modified from Patanjali and Raza 2013 Raza 2013;; Jones and Burges 1997 Burges 1997)) Formulation
Abbreviation
Formulations diluted in water Emulsifiable concentrate EC Water-in-oil emulsion EO Oil-in-water emulsion EW Suspension concentrate SC Capsule suspension CS Soluble concentrate SL Water-soluble powder SP Water-soluble granule SG Tablet TB Briquette BR Wettable powder WP Water-dispersible granule WG Formulations diluted with organic solvents Oil-miscible liquid OL Oil-miscible flowable OF Oil-dispersible powder OP Formulations applied undiluted Dustable powder DP Encapsulated granule CG Microgranule MG Electro-chargeable liquid ED Spreading oil SO Ultralow volume liquid UL Ultralow volume suspension SU Granule GR Seed treatments
Powder for dry seed treatment Flowable concentrate Solution for seed treatment Coated seed Water-dispersible powder Miscellaneous Bait concentrate Bait
Fe F eatures Emulsion formed when added to spray tank Preformed emulsion Preformed emulsion Suspended insoluble AI AI contained in capsules Used for water-soluble AI Powder soluble, but may contain inert ingredients Used for water-soluble AI Used for portable water-soluble AI Controlled-release formulation Typically consist of AI, clay carrier, and surfactants AI dispersed, but not dissolved, in water AI dissolved in organic solvent Suspension in organic liquid Powder to be applied in oil AI carried on free-flowing powder Controlled-release granule Diameter below O.6 mm Used with electrostatic spray equipment Applied to water surface Applied through UL V sprayers As above Applied to soil and water
DS FS LS PS SS
Liquid suspension
CB RB
Bait diluted before application
then amended with the product of sorghum ferment mentat atio ion. n. Isol Isolat ates es of T. harz harzia ianu num m and G. rose roseum um were were allo allowe wed d grow growin ing g on thes thesee granules for 7 days. These granules were allowed to air-dry followed by incubation before application in R. solani -infested soil. Trapping of antagonis onisti ticc micr microo oorg rgan anis ism m in calc calciu ium m algi algina nate te gran granul ules es,, also also know known n as pril prill, l, has has been been used used widely widely used (Connick (Connick 1988). 1988). Though most of the commercially available alginates are derived from kelp, other organisms are also reported to prod produc ucee algi algina nate tes. s. Algi Algina nate tess prod produc uced ed by
Azotobacter vinelandii can be used in place of kelp kelp algina alginate te for the produc productio tion n of microb microbial ial formulat lation to control plant pathogens (De Lucca et al. 1990). 1990). Furthe Furtherr resear research ch may lead to less costly alginates. Sodium alginates showing great variation in viscosity and purity are are avai availa labl blee comm commer erci cial ally ly.. More More gran granula ular r sodium sodium algina alginates tes like like Kelgin Kelgin HV, Kelgin Kelgin,, and sodi sodium um algi algina nate te IG-3 IG-350 50 are are easi easier er to hand handle le than the more powdery alginic acids. Daigle and Cotty (1995 (1995)) reported that 5 % gluten from wheat grains improved the
2
Formulation Technology of Biocontrol Agents: Present Status tus and Future Prospects
performance of a toxigenic Aspergillus flavus but that concentrations were very complex to process. cess. The final final formul formulated ated produc productt contai contained ned 5 % gluten, 1 % sodium alginate, and 5 % corn cob grits. Likewise, in case of Streptomyce Streptomyce s spp. formulation, clay was handy to keep in suspension, ion, form formul ulat ateed in algi algin nate ate with with carr carrie ier r polyamide.
2.3 2.3
Bioc Biocon ontr trol ol Prod Produc ucts ts Containing Fungi and Their Formulations
Plethora of antagonistic fungi and their products have been registered as commercial biopesticides glob global ally ly (Tab (Table le 2.3 2.3)). Different types of formulations of these antagonists are currently available in markets and are successfully used against against various various phytopath phytopathogen ogens. s. Trichoderma spp., Gliocladium spp., Coniothyrium , and nonpathogenic strains of fungal genus are available as different formulations. Vari Variou ouss biop biopes esti tici cide de prod produc ucts ts (Bin (Binab ab-T -T,, Bio-Fung Bio-Fungus, us, Supresivit Supresivit,, RootShield RootShield,, T-22HB, T-22HB, T-22 T-22G, G, Tric Tricho hode dex, x, Tric Tricho hose seal al,, Tric Tricho hope pel, l, Tricho Trichodow dowels els,, BioMax BioMax,, BioVam BioVam,, Tricho Trichojec ject, t, Trichoderma 2000) contain Trichoderma spp. to control a variety of pathogens, including Fusarium, Botrytis, Gaeumannomyces , Rhizoctonia , Pythium, Sclerotinia , Verticillium, Sclerotium , and wood-rot fungi. Trichoderma product formulati lation on vari varies es cons consid ider erab ably ly.. Co Comb mbina inati tion on of Tricho Trichoder derma ma viride viride and T. harz harzia ianu num m is formulated differently, for example, as a pellet for soil (Trichopel), as dowels for injecting in woods (Trichodowels), as a wettable powder for resusp resuspens ension ion to apply apply on wounds wounds with brush brush (Trichoseal), and as a wettable powder in syringe (Trichoject). Bio-Fungus is available as a granule, as crumbles for soil incorporation, and as wetta ettabl blee powde owderr impr impreg egna nate ted d in stick ticks. s. Gliocladiu Gliocladium m virens virens formulation is available in alginate alginate prill under under the trade name SoilGard SoilGard and effectively used against R. solani and Pythium spp. Nonpat pathogenic strain of Fusarium oxysporum has has been been used used agains againstt pathog pathogenic enic F. oxysporum and Fusarium Fusarium moniliforme moniliforme on
39
carnation, tomato, basil, and cyclamen. Commercial cially ly avai availa labl blee prod produc ucts ts of nonp nonpath athog ogen enic ic F. oxysporum , namely, Biofox C, are formulated as alginate prill or dust, and Fusaclean is available able as microg microgran ranule ule.. Aspire Aspire contai containin ning g yeast yeast Candida Candida oleophila oleophila, is form formul ulate ated d as wetta wettabl blee powder for postharvest application in citrus and pome fruit to control Penicillium spp. and Botrytis. Ampelomyces quisqualis , parasite of powdery mild mildew ew fung fungi, i, is comm commer erci cial ally ly avai availab lable le as water-disp water-dispersib ersible le granule granule (AQ10) (AQ10) and applied applied to the leaves leaves of apples apples,, grapes grapes,, strawb strawberr erries ies,, tomatoes, tomatoes, cucurbits, cucurbits, and ornamental ornamentals. s. Pythium oligandrum has been formulated as granule or powder for use in seed treatment for management of pathogenic Pythium spp. (Jones and Burges 1997). 1997 ). Lewis and Lumsden (2001 (2001)) prepared a solid matr matrix ix form ormula ulation tion of Gliocladium and Trichoderma using wheat bran and vermiculite effective against R. solani . Formulated product was applied at a rate of 1.0 % (w/w). The product was was tes tested, ted, and and a signi ignifi ficant cant red reducti uction on in damping-off of pepper seedlings was observed. Efficacy of Trichoderma Trichoderma spp. against Sclerotinia sclerotiorum causi causing ng sunfl sunflowe owerr head head rot rot was was evaluated in the field. Trichoderma formulation contained viable hyphal fragments, Trichoderma conidia, milled corn kernels, and industrial talc. Sunflower heads were infected with S. sclerotiorum sclerotiorum after 2 days of the first delivery Trichoderma formulation by honeybees. Head of Trichoderma rot incide incidence nce was reduce reduced d in sunflow sunflower er when when 100 g formulation was carried by honeybees in a 10 h per day period (Escande et al. 2002 al. 2002). ). An invert emulsion based on soybean and coconut oils offered the lowest viscosity (27 0.81 cps) and most stable emulsion layer (93 % v/v) for formulating T. harzianum conidia (Batta (Batta 2004). 2004). In this formulation, 36 months of shelf life of conidia was recorded with 50 % declined viability at 20 1 C after 5 months. Botrytis sporulati lation on on the the frui fruitt lesi lesion on surf surfac acee was was also also inhibited after 10 days of inoculation. Biocontrol potential potential of Trichoderma isolated isolated from rhizorhizosphe sphere re of Musa sp. sp. was was evalu evaluat ated ed agai agains nstt F. oxysporum in vitro. Among different isolates, T. harzianum Th-10 was found most significant
40
C. Keswani et al.
Table 2.3 Some commercially available fungal biocontrol products Organism
Product name
Di Disease against used
Formulation
Application
Ampelomyces quisqualis
AQ10 Biofungicide
Powdery mildew
Spray
Candida oleophila Trichoderma harzianum, T. polysporll
Aspire
Penicillium spp. Botrytis spp.
Waterdispersible granule Wettable powder Wettable powder
Fusarium oxysporum
BiofoxC
Trichoderma spp.
Bio-Fungus Sclerotinia, Rhizoctonia solani, Pythium spp., Fusarium, Verticillium, Phytophthora
Coniothyrium minitans Myrothecium verrucaria (killed cells)
Contans DiTer iTeraa
Sclerotinia sclerotiorum and S. minor Roo Root-kn t-kno ot, citr citrus us cyst cyst,, stub stubby by root, sting, lesion, and burrowing nematodes
Fusarium oxysporum (nonpathogenic)
Fusaclean
F. oxysporum
Phlebia gigantean T. harzianum strainT-22 Gliocladium virens
Rotstop
Heterobasidion annosum
Bina Binabb-T T
Trichodex
Trichopel, Trichoject, Trichodowel Trichoseal Trichoderma sp. Trichoderma 2000 Sardar Eco T. harzianum NBRI-1055 Green Biofungicide Tricha T. harzianum NBRI-1055 T. harzianum and T. viride
Path Pathog ogen enic ic fung fungii caus causin ing g wilt wilt,, take-all, root rot, internal decay of wood products, and decay in tree wounds F. oxysporum, F. moniliforme
Root Shield Pythium spp., R. solani, Fusarium spp. Soil Gard Rhizoctonia solani, Pythium (formerly spp. GlioGard) Supr Supres esiv ivit it Vari Variou ouss fung fungii T-22G Pythium spp., R. solani, 19, T-22 HB Fusarium spp., Sclerotinia homeocarpa
T. harzianum T. harzianum and T. viride
T. harzianum
Botrytis cinerea, Colletotrichum spp., Plasmopara viticola, Armillaria, Botryosphaeria, Fusarium, Chondrostereum
Dust or alginate granule Granules, wettable powders, sticks, and crumbles
Postharvest application to fruit as drench, drip, or spray Postharvest application to fruit as drench, drip, or spray
Seed treatment soil or incorporation Applied after fumigation, incorporated incorporated in soil sprayed or injected
Spray Wettable powder, emulsifiable liquid or granule Spores, microgranule Spores in inert powder Gran ranules ules Granule (1 10 6 cfu g1) Granules or dry powder (both at 1 107cfu g1) Wettable powder
R. solani , S. rolfsii, Pythium sp. Pythium sp., R. solani, S. rolfsii Wettable powder Pythium sp., R. solani, S. rolfsii Wettable powder
In drip to rock wool; incorporate in potting Spray, chain saw oil Mix Mix with with soil soil or pott pottin ing g medium Granules are incorporated in soil or soilless growing media prior to seedlings Granules Granules added in furrow with granular applicator, by broadcast application to turf, mixed with green house soil Spray
Incorporated into soil or potting medium Spray
Spray
2
Formulation Technology of Biocontrol Agents: Present Status tus and Future Prospects
in pathogen pathogen inhibition inhibition.. Five organic substrates substrates includ including ing rice rice chaffy chaffy grain, grain, rice rice bran, bran, banana banana pseudostem, farmyard manure, and dried banana leaf leaf were were tested tested for the mass mass produc productio tion, n, and drie dried d bana banana na leaf leaf was was foun found d the the best best carr carrie ier r material for T. T. harzianum growth. Dried banana leaves were colonized within a few days by strain Th-10 and produced propagules of high density (4.6 1032 cfu g1 of leaf). leaf). Furthermor Furthermore, e, addition of jaggery (10 % w/v) to the dried leaves enhances the growth of T. T. Harzianum, and more than 6 months of surviva ival was recorded (Thangavelu et al. 2004 al. 2004). ). Trichoderma Trichoderma atroviride atroviride isol isolat atee C52 was was observed on onion roots when inoculated in soil with various formulations (McLean et al. 2005 al. 2005). ). Pellet formulation product maintained the con Trichoderma up to 105 cfu g1soil centration of Trichoderma in comp compar aris ison on to 101 cfu cfu and and 104 g1 soil concen concentra tratio tions ns that that were were mainta maintaine ined d by seed seed coating and solid substrate formulations, respectively. Trichoderma isolate C52 was inoculated into Sclerotium cepivorum -infested soil as both solid substrate and pellet formulations, and no difference was observed in disease control, but more healthy plants were observed in the pellet treatment. Increased root and shoot lengths, plant height, and dry weight were recorded after treatment of plants with T. viride formulated in talc. Application of T. T. viride formulation also resulted in a significant reduction of sheath blight caused by R. solani (Mathivanan et al. 2005 al. 2005). ). New carrier ier form formu ulati latio on was dev develop eloped ed by usin using g T. harzianum M1, and resistance to carbendazim show showed ed inhi inhibi bito tory ry effe effect ct agai agains nstt Pythium aphanidermatum. Different formulations including lignite, talc, wettable powder, lignite + fly ash-based powder formulation, bentonite paste, gelatin-glycerin gel, and polyethylene and glycol paste were developed for seed treatment. Shelf life of the microbial formulations was assessed at 24 C for 9 months. Up to 74 % reduction in disease incidence was recorded when Trichoderma formul formulati ation on was applie applied d as seed seed treatment. Additionally, enhanced plant biomass under field and greenhouse conditions was also reco ecorde rded (Jay Jayaraj araj et al. al. 2006). 2006). Pell Pellet etiz ized ed formulations of kaolin clay and wheat bran in
41
an algina alginate te gel contai containin ning g conidi conidia, a, fermen fermentor tor biomass, biomass, or chlamydosp chlamydospores ores of G. virens virens and Trichoderma spp. spp. were were prep prepare ared d (Lew (Lewis is and and Papavizas 2007 Papavizas 2007). ). Higher population densities of Trichoderma and Gliocladium were observed observed when soil is incorporated with alginate pellets containing containing chlamydosp chlamydospores ores rather rather than condia condia and bran rather than kaolin as the carriers. A new Trichoderma asperellum formulation was develo developed ped using using soybe soybean an oil disper dispersio sion. n. Complete inhibition of Phytophthora Phytophthora megakarya causing cacao black pod disease was recorded when when form formul ulat atio ion n was was appl applie ied d to the the pods pods.. Ninety percent prevention of infection in treated pods was recorded after 1 week, and 50 % reduction after 3 weeks was recorded when formulation was sprayed on cacao clones susceptible to P. megakarya. The formulations showed a significant effect in disease management (Mbarga et al. 2014 al. 2014). ). Talc-based formulation of a novel T. virid viridee, BHU-29 BHU-2953, 53, succe success ssfull fully y contro controlled lled damping-off of chili caused by P. aphanidermatum and tomato wilt caused by F. oxysporum. Significant reduction in diseases was observed when T. viride formulated as 2 % wettable powder was applied to the seeds and furr furrow ow (Sin (Singh gh et al. al. 2014). 2014). Formul Formulatio ations ns of Trichoderma with with mixt mixtur uree of 1 % w/v w/v Su Sure re-1 Jell, 1 % w/v PDB, and 0.3 ml L of the surfactant tant Break BreakThr Thru u 100SL 100SL (BT) (BT) and an invert invert oil emulsion of 50 % v/v corn oil, 2.5 % w/v lecithin, and 0.5 % w/v PDB (COP) were evaluated against frosty pod rot pathogen Moniliophthora roreri. T. harzianum DIS 219f and Trichoderma ovalisporum DIS DIS 70a 70a were were appli applied ed (180 (180 ml tree tree1, 2.46 107 conidia ml1) in the field. COP/DIS70a formulation resulted in a maximum increase in yield as compared to other treatments (Crozier et al. 2015 al. 2015). ).
2.4 2.4
Bioc Biocon ontr trol ol Prod Produc ucts ts Containing Bacteria and Their Formulation
Seve Several ral bact bacter erial ial spec specie iess havi having ng bioc biocon ontr trol ol potential have been formulated and are commercial cially ly avail availab able le (Tab (Table le 2.4 2.4). ). Nonpat Nonpathog hogeni enicc
42
C. Keswani et al.
Table 2.4 Some commercially available bacterial biocontrol products Biocontrol agent
Trade name
Pseudomonas syringae ESC 10
BioSave10
P. syringae ESC 11
BioSave11
P. fluorescens A506 P. fluorescens NCIB B. subtilis
Blight Ban A 506 Conquer
Agrobacterium radiobacter
GalltrolA13
Pseudomonas cepacia A. radiobacter
Intercept R. solani, Fusarium spp., 14 Pythium sp. Nogall, A. tumefaciens Diegall 16 PSSOL 12 P. solanacearum
P. solanacearum (nonpathogenic) P. fluorescens NCIB 12089 B. subtilis B. cepacia-type Wisconsin M36
Epic II
Target pathogen
Formulation
Application
B. cinerea, Penicillium spp., Mucor piriformis, Geotrichum candidum B. cinerea , Penicillium spp., M. piriformis, G. candidum
Wettable powder
Frost, Erwinia amylovora
Wettable powder
Postharvest application to fruit as drench, dip, or spray Postharvest application to fruit as drench, dip, or spray Drench dip or spray
P. tolaasii
Aqueous biomass suspension Dry powder (5.5 1010 spores g1)
R. solani, Fusarium, Alternaria spp., and Aspergillus spp. Crown gall disease A. tumefaciens
Wettable powder
Petri dishes with pure culture grown on agar (1.2 1011 cfu plate1) Wettable powder
Spray Added to a slurry, mix with a chemical fungicide Root dips, drench
Drench dip or spray
Washed plates, culture suspension
Root dips
Victus
R. solani, Fusarium spp.
Aqueous suspension
Spray
System3
S. rolfsii, S. sclerotiorum
Dust
Blue Circle
Fusarium, Pythium, spiral, lesion, lance, and sting nematode
Peat Peat carr carrie ierr or liqu liquid id
Seed treatment in planter box Seed eed trea treatm tmen entt or drip irrigation
A. radiobacter is commercially available under to the chick pea seed, the total shelf life of the various trade names such as Nogall, Galltrol-A, bacteria was recorded at 180 days (Vidhyasekaran Diegall, Diegall, and Norbac 84C. Bacteria Bacteria are generally and Muthamilan 1995 Muthamilan 1995). ). Peat-based formulation of suspended in non-chlorinated water and applied P. fluorescens strains PfALR2 was developed and as dips and sprays to cuttings and stems or as soil assessed for root treatment, seed treatment, foliar Pseudomonas syringae syringae formulate drench. Pseudomonas formulated d as spraying, spraying, and soil application. application. All four treatments wett wettab able le powd powder erss is comm commer erci ciall ally y avai availa labl blee in combin combinatio ation n result resulted ed in the significa significant nt control control of under under trade trade names names Bio-Sa Bio-Save ve 10 and Bio-Sa Bio-Save ve sheath blight in greenhouse greenhouse condition (Rabindran (Rabindran Vidhyasekaran n 1996). 1996). P. fluorescens fluorescens strain 11 and used for postharvest application to citrus and Vidhyasekara and and pome pome frui fruitt for for mana manage geme ment nt of Botrytis, PF-1, isolated from rhizosphere of maize roots, Mucor , Penicillium, and Geotrichum. showed antagonistic potential against against R. solani solani P. fluorescens strains strains isolat isolated ed from from rhizosp rhizospher heree f. sp. sp. sasakii causing banded banded leaf and sheath blight blight of various various crops crops with with antagoni antagonisti sticc potent potential ial agains againstt of maize. Among the different carriers, talc and Fusarium Fusarium spp. were formulated as talc-based and peat maintained the population at 18.3 107 and peat-based products. In talc-based and peat-based 19.5 107 cfu cfu g1 of the bacterium, bacterium, respectively respectively,, formulations, P. fluorescens survived for a maxi- after 40 days. Significant control of disease was mum of 240 days. When formulation was applied recorded after the seed treatment with peat-based
2
Formulation Technology of Biocontrol Agents: Present Status tus and Future Prospects
formul formulatio ation n (Sivak (Sivakuma umarr et al. 2000 2000). ). Formulation Formulationss of B. subtil subtilis is AF 1 show showed ed both both plan plantt grow growth th promotion promotion and biocontrol biocontrol potential potential when prepared prepared in peat. The formulation was supplemented with A. niger myce myceliu lium, m, 0.5 0.5 % chiti chitin, n, and and orga organi nicc comcompost from cultivation of Agaricus Agaricus bisporus . Biocont contro roll pote potenti ntial al of form formul ulat ated ed prod produc ucts ts was was evalua evaluated ted against against two pathoge pathogens ns on ground groundnut nut and pigeon pea. Chitin, A. niger mycelium, and A. bisporus bisporus compost were used as supplement for improving the growth rate of B. B. subtilis AF 1A. Peat formulation supplemented with chitin when used for seed treatment demonstrated better control of wilt in pigeon pea caused by Fusarium udum and A. niger responsible responsible for crown rot disease ease in groundn groundnut ut (Manju (Manjula la and Podile Podile 2001). 2001). P. fluorescens fluorescens strains FP7 and PF1 and their consortiu sortium m were were formul formulated ated as talc-ba talc-based sed produc productt and mixed both with and without chitin were assessed assessed individually against sheath blight of rice. Significant reduction in disease incidence was recorded after after the applic applicatio ation n of formul formulated ated produc productt through through seed seed,, soil soil,, root root,, and and foli foliar ar spra spray. y. In field field as much much as 62.1 % reduction in sheath blight incidence was observed in the consortium treatment containing chiti chitin n (Com (Comma mare re et al. al. 2002). 2002). 0.1 0.1 % calc calciu ium m hydroxide hydroxide significantly significantly promoted the growth of B. amyloliquefa amyloliquefaciens ciens strain strain B190 B190 used agains againstt Botrytis Botrytis elliptica elliptica in in lily. Spraying B. amyloliquefac amyloliquefaciens iens B190 mixed with 0.05 % sodium sodium carbona carbonate, te, 0.025 0.025 % calcium calcium hydrox hydroxide ide,, or 0.025 0.025 % ammo ammoni nium um nitra nitrate te supp suppre ress ssed ed the the gray mold on lily. Concentration of adjuvant was kept below 0.1 % (v/v); carboxymethyl cellulose (CMC) and Tween 80 were effective to B. amyloliquefac amyloliquefaciens iens B190 formulation formulation against against lily gray mold (Chiou and Wu 2003 Wu 2003). ). The efficacy viride de and of talctalc-ba base sed d form formul ulat atio ions ns of T. viri P. fluorescens alone and in combination on sheath blight disease, crop growth, and yield in rice was stud studied ied in field field expe experi rime ments nts.. Appl Applic icat ation ion of form formul ulat ated ed prod produc uctt of P. fluor fluoresc escen enss and T. viride either alone or in combination resulted in increase in root and shoot lengths and plant height when compared with control. Significant redu reduct ctio ion n in shea sheath th blig blight ht incid inciden ence ce was was also also recorded after application of P. fluorescens fluorescens and T. viride (Mathivanan et al. 2005 al. 2005). ).
43
Bacillus licheniformis strain N1 exhibiting the biocontrol activity against Botrytis cinerea was formulated formulated using fermentati fermentation on of the bacterial bacterial culture in Biji medium. Wettable powder formulation of antagonist based on olive oil corn and starch was selected for evaluation of the disease control. A dose of 100-fold-diluted B. licheniformis N1E was found to be the optimum spray formulation and significantly reduced the the dise diseas ase. e. 90.5 90.5 % redu reducti ction on in dise diseas asee by formulated product was recorded in comparison to the the 77 % redu reduct ctio ion n by synt synthe heti ticc fung fungici icide de incl includ udin ing g carb carben enda dazim zim and and diet dietho hofe fenc ncar arb. b. Results of this study indicated that the olive oiland corn starch-based formulation of B. licheniformis using using liquid liquid fermentatio fermentation n will be effec effectiv tivee agai agains nstt toma tomato to gray gray mold mold (Lee (Lee et al. 2006). 2006). Ninete Nineteen en isolat isolates es of antago antagonis nistic tic Pseudomonas and twelve isolates of yeast were scre screen ened ed for for the the bioc biocon ontr trol ol acti activi vity ty agai agains nstt Colletotrich Colletotrichum um musae causing banana anthracnose. P. fluorescens strain FP7 showed a maximum inhibition of C. musae musae mycelial mycelial growth. growth. Water-in-oil formulation of P. fluorescens FP7 was formulated by adding various oils such as rice bran (28.50 %), coconut (28.50 %), and castor (28.50 %) separately to the bacterial culture and bacterial populations were reported to survive for 210 days of storage. The application of waterwater-inin-oil oil formul formulati ation on of bacter bacterium ium signifi signifi-cantly cantly reduce reduced d the diseas diseasee incide incidence nce (Peera (Peeran n et al. al. 2014). 2014). B. subtilis strain BY-2 was unable to colonize the lea leaf surface and stem in oilseed rape when applied as pellet. Populations of BY-2 declined from 10 8 CFU seed1 to 104 CFU g root1 and 1023 CFU g stem1 after 60 days. days. Signifi Significan cantt reduct reduction ion in diseas diseasee was observed when compared to control (Hu et al. 2014 al. 2014). ). Efficacy of aqueous suspension (Serenade ASO or QRD 145) and foliar sprays of wettable powder formulation (Serenade MAX or QRD 141) of B. B. subtilis QST 713 alone and in comb combin inat atio ion n with with cop copper per hydr hydro oxid xide was was invest investiga igated ted agains againstt bacter bacterial ial spot spot diseas diseasee of tomato. tomato. The aqueous aqueous suspensio suspension n of B. subtilis subtilis QST 713 alone alone signifi significan cantly tly reduce reduced d bacter bacterial ial spot on tomato foliage when compared to control. trol. The wettab wettable le powder powder of B. subtili subtiliss QST
44
C. Keswani et al.
713 alone did not reduce bacterial spot, but in B. subtil subtilis is-based -based biopes biopestici ticide, de, was used used as seed seed combinatio combination n with copper hydroxide hydroxide it reduced reduced inoculant for peanut, cotton, and beans to control dise diseas asee seve severi rity ty and and enha enhanc nced ed the the tota totall frui fruitt the the root root dise diseas ases es caus caused ed by Fusariu Fusarium m and Rhizoct Rhizoctooyield (Abbasi and Weselowski 2015 Weselowski 2015). ). nia (Mahaf (Mahaffee fee and Backma Backman n 1993 1993). ). Quantum-400 Quantum-4000, 0, a B. subtilis strain A13-based product, is commercially available as seed inoculants for peanut, and another biopesticide based on strain GB 07 named 2.5 See Seed Trea reatmen tmentt Epic Epic is availab available le for cotton cotton.. Bacter Bacterial ial BCAs BCAs activel actively y The appl applic icat ation ion of micr microb obes es to seed seed surf surfac acee colonize the rhizosphere and compete with other requires few technical considerations. Significant micr microo oorg rgan anis isms ms incl includ udin ing g path pathog ogen enss (Bis (Bisen en amou amount nt of the the inoc inocul ulum umss must must surv surviv ivee the the appl applic icaa- et al. 2015). 2015). Nons Nonspo pore re-f -for ormi ming ng bacte bacteri rium um Enterobac bacter ter cloacae cloacae showed antagonistic activity tion procedure and must have the capacity to grow Entero in vicinity vicinity of seed. seed. Since seeds are at low moisture moisture against Pythium spp. spp. caus causin ing g seed seed rot rot in cott cotton on leve levels ls for for most most of the the time time duri during ng stor storag age, e, seed seeds. s. Simp Simple le seed seed inoc inocul ulat atio ion n tech techni niqu quee with with microorganisms must have the ability to survive (CMC) as sticker in cotton seeds was carried out under low water activity. Microorganisms might successfull successfully y (Nelson (Nelson 1988). 1988). Liqu Liquid id cult cultur uree of like likewi wise se need need to be mixe mixed d with with othe otherr acti active ve E. cloaca cloacaee was applied by solid matrix priming to ingr ingred edie ien nts, ts, for for exam exampl ple, e, inse insecctici ticide dess and and tomato and cucumber seeds in combination with a fungicides. These aspects raise issues of formula- fungicide (Harman and Taylor 1988). 1988). tion stability and strain selection. Seed treatment Generally, in seed treatment, treatment, microbial formuwith beneficial microorganisms has been a prime lation is applied to seed as powder or liquid. For fluorescens and Burkholderi Burkholderia a cepacia cepacia area area of inve invest stig igat atio ion n for for many many year yearss (Bis (Bisen en example, P. fluorescens et al. 2015). 2015). Micr Microo oorg rgan anis isms ms with with vari variou ouss were applied to pea seeds with or without captan properties have been applied to seeds to perform for control of Aphanomyces Aphanomyces root rot and Pythium damping-o g-off ff (Parke (Parke et al. 1991 1991). ). Pota Potato to tube tubers rs have have various various functions, functions, including including plant growth promopromo- dampin tion, nitrogen nitrogen fixation, fixation, phosphate phosphate solubilizat solubilization, ion, been been trea treated ted with with asco ascosp spor ores es of Talaromyces and biolog biological ical contro controll of plant plant pathoge pathogens ns (Keswa (Keswani ni flavus in in a pyrop rophyllite lite carrier ier (Fravel vel al. 1985). ). In order to ensure the better adhesion et al. 2013b al. 2013b). ). BCAs are applied to the seeds for et al. 1985 protection of seed and seedlings from effects of of bioi bioino nocu culan lants ts on seed seed surf surfac ace, e, stick stickers ers are are various seed-borne and soilborne plant pathogens. added added to microbi microbial al formula formulatio tions ns for seed seed treattreatFor the succes successfu sfull biolog biological ical contro control, l, applied applied ment. To control Pythium ultimum attack in pea microorganism must grow and colonize the rhizo- and soybean seeds, PelGel has been used to treat sphe sphere re in orde orderr to prot protec ectt the the plan plant, t, thus thus the relea release se the seed with P. putida (Paulitz et al. 1992 1992). ). In of microbes from the formulation and prerequisite cucumber seed treatment, PelGel has also been harzianum (Harman 1991; condi conditio tion n for growth growth are of param paramoun ountt import importan ance. ce. used with T. harzianum 1991; Taylor The comm commer erci cial al wett wettab able le powd powder er prod produc uctt et al. 1991). 1991). Another sticker Polynox-N-I0 was used for for seed seed treatm treatmen entt with with shal shalee and and then then appl applied ied Mycostop (Kemira AgroOy, Finland), containing used 8 Streptomyces Streptomyces griseoviridis griseoviridis stra strain in K61, K61, with with 10 on bean seeds with conidia of T. harzianum, and 1 cfug is available for seed treatment. The powder efficien efficientt diseas diseasee contro controll was observ observed. ed. Various Various 1 concentrat tration ionss of plant plant gum, gum, methyl methyl cellulo cellulose, se, is usedat 5–8 g kg seeds seeds for contro controll of seed-b seed-born ornee concen and soilbo soilborne rne fungal fungal phytop phytopath athoge ogens ns in herbs, herbs, and xanthane gum with talc have been used for vegetables, vegetables, and ornamentals ornamentals (Tomlin 1994). 1994). The treatment of potato tuber with PGPR (Kloepper Schroth h 1981) 1981). At 40 C, popu popula lati tion on of shel shelff life life of form formul ulat atio ion n is esti estima mate ted d to be 6 mont months hs and Schrot at 8 C or for 12 months at 12 C and must be rhizobacteria in talc and 20 % xanthane gum did stored in airtight containers. Seed inoculation with not decline for 2 months and resulted in better bacte acteri riaa to pro protect tect the the seed seed from from soilb oilbor orn ne incr increa ease se in plan plantt deve develo lopm pmen entt in field field pota potato to.. path pathog ogen enss and and prom promot otee plan plantt has has been been well well However, rhizobacteria did not last longer in forinvest investiga igated ted (Merri (Merriman man et al. 1974). 1974). Kodi Kodiak ak,, a mula mulati tion on cont contai aini ning ng gum gum traga tragaca cant nth h or gum gum
2
Formulation Technology of Biocontrol Agents: Present Status tus and Future Prospects
45
kara karaya ya.. CMC CMC was was appl applie ied d with with micr microb obia iall agen agents ts to seeds to control R. solani. One percent CMC with clay carrier carrier was applied applied to Verticillium Verticillium biguttatum biguttatum and other biocontrol agents to potato tubers (Jager and Velvis 1985 Velvis 1985). ). Various antagonists have been applied to treat the sugar beet seeds using either gum xanthan or methylcellulose in combination with a neutralized neutralized talc or peat carrier (Suslow (Suslow and Schroth 1982 Schroth 1982). ). Similarly, to control take-all disease ease in wheat, wheat, surf surfac acee-di disi sinf nfes este ted d seed seedss were were coated with P. fluorescens fluorescens in combination with 1 % methylcellulose (Weller and Cook 1983 Cook 1983). ). In seed treatment of Chinese aster with T. flavus, a polymer binder was used in quartz flour to pellet seeds seeds (Nagtza (Nagtzaam am and Bollen Bollen 1994 1994), ), and antagon antagonist ist T. flavus was isolated from seeds after 17 years. Strains Strains of E. E. cloacae cloacae and Trichoderma were delivdelivered to cucumber and tomato seeds through solid matrix matrix primin priming g (Harman (Harman and Taylor Taylor 1988 1988). ). During During priming seeds are brought to a certain moisture level just below the required level for germination and then mixed with moistened Trichoderma or E. cloacae cloacae, shale, sphagnum moss, or bituminous coal. The seeds and carriers are then mixed with water and incubated before planting.
of the the micr microo oorg rgan anis ism m prio priorr to inoc inocul ulat atio ion n in field, (2) poor survival of the organism in the environment, and (3) low quality of the microbial product itself. Success of biopesticides depends on the delivery of viable, active microorganisms in high numbers to the field which requires highqualit quality y inocul inoculant ants. s. The carrie carrierr substr substrate ate is the most critical part of the microbial formulation, and it must be capable of supporting high numbers of microbe. Carriers are inert ingredients, and they do not have biocontrol potential; however, they can affect the efficacy of the product and shelf life of microorganism (Table 2.5 (Table 2.5). ). Better surviv survival al of Pseudomonas spp. spp. on minera minerals ls with small particle size, such as zeolite, montmorillonite, and vermiculite, than on minerals with larger particle size such as talc, pyrophyllite, and kaolinite was observed during storage at 20 C (Dan (Dandu dura rand nd et al. al. 1994) 1994). Backman and RodriguezRodriguez-Kabana Kabana (1975 1975)) compar compared ed diatom diatomaaceous earth and attapulgus clay for their various physic physical al proper propertie tiess includ including ing waterwater-hol holdin ding g cap capacit acity y and and stre trength ngth afte afterr autoc utocla lav ving. ing. Attapulgus clay granules swelled in water and lost lost their their integr integrity ity after after autocl autoclavin aving, g, wherea whereas, s, diatom diatomace aceous ous earth earth granul granules es did not swell swell in water and remained intact after autoclaving thus making it to absorb a molasses-based medium for 2.6 2.6 Carr Carrie iers rs and and Adju Adjuva vant nt Used Used delivery delivery of T. harzianum harzianum to soil. Wheat branin Microbial Formulations perlit perlitee mixtur mixturee and poplar poplar bark bark compos compostt have have been used as carrier for nonpathogenic Fusarium Vari Variat atio ions ns in eff effecti ective vene nesss of micro icrobi biaal strains mixed with soil that induced resistance to formul formulati ations ons from from benefi beneficial cial microo microorga rganis nisms ms F. oxysporum f. f. sp. dianthi (Garibaldi are credited to three main causes: (1) presence et al. 1987). 1987). Fine-g Fine-grou round nd tree tree barks barks have have also also
Table 2.5 Various adjuvants used in microbial formulations Adjuvant
Type
Function
Oils
Mineral oils Crop oil Wetting agents Spreaders Penetrants Emulsifiers Dispersants Anti-flocculating agents Cosolvents Coupling agents Humectants
Improve uptake, photostability
Surfactant
Stabilizing agents
Solvents Hygroscopic agents
Improve spreading, wetting, or dispersion
Maintain stability during application
Maintain AI in solution Prevent premature drying of deposit
46
C. Keswani et al.
been used as a carrier (Stack et al. 1988 al. 1988). ). Peat is generally generally used as carrier carrier for Rhizobium Rhizobium spp. and is also useful for soil applications and seed coating of biocontrol agents. Huber et al. (1989 ( 1989)) used fine-ground peat with a methylcellulose sticker for wheat seed treatment with bacteria for control Gaeumannomyces graminis . Alder bark has of Gaeumannomyces been used as a carrier to apply T. flavus to potato seed pieces (Keinath et al. 1990 al. 1990). ). One of themec the mechan hanis isms ms invol involved vedin in biocon biocontro troll is the production production of hydrolytic hydrolytic enzymes by the antagantagonistic microorganism. The amount and type of nutrients in the formulation must allow ample production of hydrolytic enzymes (Stack et al. 1988 1988). ). Increas Increased ed molar molar concentr concentratio ations ns of carbon carbon and nitronitrogen sources (0.02–0.18 M maltose and 0.006–0 0.006–0.024 .024 M arginine arginine)) and increas increased ed carbon/ carbon/ nitrogen ratios (12:1–80:1) enhanced the proliferaproliferaThielaviopsis basicola and Trichoderma tion tion of Thielaviopsis spp. on lignite granules (Stack et al. 1987 al. 1987). ). Likewise, T. flavus was formulated with eight different orga organic nic carrie carriers rs and used used again against st Verticillium dahliae on eggplant, and a maximum inhibition of patho pathoge gen n was recor recorded ded in treatm treatmen ents ts with with the highest carbon/nitrogen carbon/nitrogen ratios (159:1 for pyrophylpyrophyllite lite,, 97:1 97:1 for for corn corn cobs cobs)) (Fra (Frave vell et al. al. 1985 1985)). As new information on mechanism of biocontrol revealed, it may be possible to express desirable biocontrol trait traitss by manipu manipulat latin ing g nutri nutrient ent comp compos ositi ition on of formulations. For example, biocontrol potential of G. virens virens GL-2 GL-21 1 depe depend ndss on the the form form of nitr nitrog ogen en in the formulation. Alginate prill of G. virens with wheat bran as a carrier resulted in significant control of Sclerotiu Sclerotium m rolfsii rolfsii in compa comparis rison on to vermi vermicuculite plus wheat bran (Ristaino et al. 1994 al. 1994). ).
2.7 2.7
Basi Basicc Info Inform rmat atio ion n Requ Requir ired ed for Microbial Product Registration
Certain information about the various parameters is prerequisite for the biopesticide registration. These parameters include microorganism strain specifications, cfu count of microorganism used, target target microo microorga rganis nism, m, moistu moisture re conten contentt of the product, type of formulation, and technical bulleti letin/ n/pr prod oduc uctt profi profile le.. Stra Strain in spec specifi ifica cati tion onss
include information on genus and species, rhizosphere competence, biological control potential, grow growth th promo romoti tion on poten otenti tial al,, and and growt rowth h parameters like pH and temperature.
2.7.1 2.7.1
Strain Strain Specif Specifica icatio tions ns
Selection of potential antagonist strain under field and lab condi conditio tions ns ensur ensures es the the effec effectiv tivee and cons consisistent performance of bioagent in field. Screening of effect effective ive strai strain n can be done done in differ different ent ways: ways: sele electi ction of poten otenttial ial strain ain in relati lation on to phytopa phytopathog thogens ens,, screenin screening g of isolates isolates with high biotechn biotechnolog ological ical applicati application, on, or search search for economeconomical viable substrates which are suitable for mass production of bioagent (Singh et al. 2003 al. 2003,, 2006 2006). ).
2.7.2 2.7.2
Shelf Shelf Lif Life e and Storag Storage e
For commercialization of a microbial product in the market, it is essential that it has long shelf life and can be stored at room temperature. It has been been sugg sugges este ted d that that shel shelff life life of a mini minimu mum m 18 month onthss is accep ccepta tabl blee for comm commer erccial ial microorganism-based product. Storage at room temp temper erat atur uree is an esse essent ntia iall cond conditi ition on as the the farmers cannot afford the equipment to keep the produc productt at any temper temperatu ature. re. Accord According ing to the rece recent nt guid guidel elin inee of the the Cent Centra rall Inse Insect ctic icid idee Boar Bo ard d and and Regi Registr strat atio ion n Co Comm mmit itte tee, e, the the data data requ requir ired ed for for clai claimi ming ng 1-ye 1-year ar shel shelff life life of the the product is for 15 months for talc-based formulation, i.e., the microbe should remain viable for 15 months.
2.7. 2.7.3 3
CFU CFU Coun Countt
According to the Central Insecticide Board and Regi Regist stra rati tion on Co Comm mmit ittee tee guid guideli eline ne,, colo colony ny-forming unit (cfu) count should not be less than 2 106 spores ml1 or g1 on selective media (SM) for antagonistic fungi, and CFU count on sele select ctiv ivee medi medium um shou should ld be a mini minimu mum m of 8 1 1 1 10 ml or g for antagonis antagonistic tic bacteria bacteria (Singh 2012 (Singh 2012). ).
2
Formulation Technology of Biocontrol Agents: Present Status tus and Future Prospects
47
faci facilit litie iess which which are are gene genera rally lly not not avail availab able le to most producers, sellers, and farmers. Shelf life is result of combini combining ng severa severall factors factors includin including g The pathog pathogeni enicc contam contamina inants nts such such as Salmo- a result production tion technol technology ogy,, materia materiall used used as carrier carrier nella, Shigella , and Vibrio should not be present. produc packaging, ing,and and transpo transport. rt. The mass mass produc production tion Other microbial contaminants must not to exceed and packag of significa significant nt number numberss of viable, viable, efficien efficient, t, and 1 104 counts ml1or g1. stable propagules of the microorganism is a prerequ requis isit itee in biop biopes estic ticid idee deve develo lopm pmen entt (Sing (Singh h et al. al. 2004b). 2004b). Submerg Submerged ed fermenta fermentatio tion n system system 2.7.5 2.7.5 Moistu Moisture re Conten Contentt has been traditionally followed by the producers over over the the solid solid subs substr trate ate ferm fermen entat tatio ion n becau because se of its Maxim aximum um mois moistu ture re cont conten entt of the the prod produc uctt cost-effectiveness and easy technology (Churchill should not exceed more than 8 % for dry formu1982;; Stowell 1991). 1982 1991). However, new large-scale lati lation on of fung fungii and and 12 % for for bact bacter eria ia (www. production systems are required for bacteria and cibrc.nic.in/2.1.22011.doc). cibrc.nic.in/2.1.22011.doc ). fungi that do not like to produce spores in liquid media. media. Unfortun Unfortunatel ately y fermen fermentati tation on system systemss for mass production based on solid substrates are not freq freque uent ntly ly avai availa labl blee (Con (Conni nick ck et al. al. 1990). 1990). 2.8 2.8 Cons Constr trai aint ntss in the the Prod Produc ucti tion on Submerged Submerged fermentation fermentation methods methods are generally generally of Microorganism-Based well adopted adopted for the mass mass produc production tion of second secondary ary Biopesticides metabolites, metabolites, antibiotics, antibiotics, organic organic acids, and bacteria; howe howeve ver, r, it is not not suit suitab able le for for the the prod produc ucti tion on of The main reasons for slow growth of microbial- ria; based biopesticide biopesticide industry industry include include inconsisten inconsistentt viable filamentous fungi. Therefore, selection of performance of final product in field condition, the cost-effective fermentation technology for the short shelf life of microorganism in formulation, mass production viable and efficient propagules is possibilities of contamination with other plant and a matter of concern. Choice of carriers and adjuhuman human pathoge pathogens, ns, lack of suitab suitable le applicat application ion vant used in the formulation is another technical problem lem in the the deve develop lopme ment nt of stab stable le and and effe effect ctiv ivee technology, small market size, and lack of proper prob knowle knowledge dge about about the biopes biopestici ticides des in farmers farmers.. biop biopes esti tici cid des. One One of the the majo majorr goals in Research should be focused on the development formul formulatin ating g biopes biopestici ticides des is to maintain maintain the viabilviabilof superior formulation to protect the reliability of ity and effectiveness of the active ingredient for a the product, because a single failure will jeopar- possib possible le durati duration, on, prefer preferably ably 2 years. years. After After the dize the whole trade’s reputation. Production of production production of microorganis microorganism, m, the main challenge biopes biopestici ticides des is a long long proces processs which which include includess faced by producers in formulation development is selection of suitable strain for formulation, mass the shelf life of microorgani microorganism sm during the storage production of selected strain, screening of micro- peri period od.. If the the prod produc uctt carr carrie iess less less numb number erss of activ activee organi organism sm for suitabl suitablee carrier carrier for formula formulatio tion, n, ingredients due to shorter shelf life, the overall assessment of the shelf life of microorganism in performance of the formulation will be affected. selected formulation, and product efficacy in field condition. Several reports on contamination and low population of microorganism in biopesticides 2.9 2.9 Futur uture e Pro Prospe spects cts being sold in the market were registered (Singh et al. al. 2004a 2004a;; Alam Alam 2000 2000;; Aror Aroraa et al. al. 2010 2010). ). Due Due to The necessi necessity ty for more safe safe produc products ts for plant low microb microbial ial count, count, it is obviou obviouss that that their their perfo perforr- diseas diseasee managem management ent prompt promptss an inclinat inclination ion of mance in the field is inconsistent and poor. The microbial biopesticide formulations with efficient unpr unpred edict ictab able le seas season onal al natu nature re of the the exis existin ting g antagonism and good stability. Biopesticides give demand needs capable storage for biopesticides. eco-friendly eco-friendly alternatives alternatives to synthetic synthetic pesticides, pesticides, The storag storagee requir requires es sophisti sophisticate cated d and specia speciall yet yet they they conf confro ront nt vario various us diffi difficu culti lties es in their their
2.7.4
Pathogen Pathogenic ic Contamin Contamination ation
48
production production,, formulation, formulation, and application. application. Since biopes biopestici ticide de generall generally y contai contains ns live organi organism, sm, maxi maximu mum m care care is need needed ed to main mainta tain in the the micr microb obia iall population and efficacy from beginning to the end use. use. Study Study of their their formul formulatio ation n and produc production tion could enormously help in the commercialization of biopesticides. It appears to be that biopesticides will will have have a more more exte extens nsiv ivee use use in the the futu future re as thei their r application techniques enhance as less expensive iner inertt mate materi rial alss are are reco recogn gniz ized ed for for diff differ eren entt form formul ulat atio ions ns.. Intr Introd oduc ucti tion on of new new adjuv adjuvan ants ts showed showed significa significant nt increas increasee in activity activity of microbe microbess and proposed a new area of research. Selection of proper formulation may enhance the product stabili bility ty,, larg larger er shel shelff life life,, and and perf perfor orm mance ance of microbes in field conditions. Biopesticides offer a more balanced plant protection product application, and in the future formulation products should have more balance between production cost and efficiency (El–Sayed 2005 (El–Sayed 2005;; Rao et al. 2007 al. 2007;; Glare et al. 2012 al. 2012;; Khater 2012). 2012). Development related to the formul formulatio ation n type type would would possib possibly ly shift shift from dusts to granules, from suspension concentrates and and wett wettab able le powd powder erss to wate waterr-di disp sper ersi sibl blee granules, granules, and from single microorgani microorganism-bas sm-based ed product to microbial consortium-based formulation. With the advances in nanotechnology science, different new microbial formulations such as nan nanosusp suspen enssion, on, nanoemu emulsion, and and nanocapsule suspension with superior efficiency will be released in the market (Rao et al. 2007 2007;; Ghormade et al. 2011 al. 2011;; Glare et al. 2012 al. 2012). ). SignifiSignificant cant adva advanc ncem emen entt has has been been made made in the the prod produc uctio tion n of new new form formul ulati ation on prod produc ucts ts and and appl applic icat ation ion methods; however, there is still much work to be done. For further research to improve production and and appl applic icat atio ion n tech techni niqu ques es,, scie scien ntist tist and and researchers are likely to provide safe and effective products for plant disease management. Eco-friendly and safer biopesticides play an important role in modern agriculture; however, thei theirr majo majorr draw drawba back ckss have have led led to the the use use of nano nanote tech chno nolo logy gy in agri agricu cultu lture re (Mis (Mishr hraa and and Singh 2015). 2015). Silver nanoparticles (AgNPs) are the most frequently used metallic nanoparticles in various sector including ing agricultu lture (Jo et al. 2009; 2009; Kim et al. 2012; 2012; Mish Mishra ra et al. 2014). 2014). Many workers have reported the
C. Keswani et al.
antimicrobial activity of AgNPs against a vast range range of phytop phytopath athoge ogens. ns. A nanosi nanosized zed silica silica-silver silver partic particle le formul formulatio ation n was develo developed ped and showed significant antimicrobial activity against a wide wide rang rangee of phyt phytop opat atho hoge gens ns incl includ udin ing g Colletotrichum sp., Pythium sp., P. syring syringae ae, Xanthomonas compestris , etc. (Park et al. 2006 al. 2006). ). Biosynthesized AgNPs of bacterium Serratia sp. BHU-S4 showed significant antifunBipolaris is soroki sorokinia niana na gal gal acti activi vity ty agai agains nstt Bipolar (Mishra et al. 2014 al. 2014). ). Recove Recoverin ring g the real real connec connectio tions ns of AgNPs AgNPs with agroecosy agroecosystems stems including including soil, soil biota, biota, and plants and their poisonous quality level can be infe inferr whet whethe herr AgNP AgNP appl applic icati ation on coul could d be helpfu helpfull for agroec agroecos osyst ystems ems.. Consid Consideri ering ng the elements deciding destiny, transport, portability, and and pois poison onou ouss quali quality ty of AgNP AgNPss in soil soil,, it is additionally accepted that the size of the poisonous ous qual qualit ity y of thes thesee part particl icles es coul could d real really ly be evad evaded ed by cont contro rolli lling ng them them and and henc hencef efor orth th requires more profound exploration. Specifically, rese resear arch ch conc concen entr trat atin ing g on this this meth method odol olog ogy y utiliz utilizing ing biosyn biosynthe thesiz sized ed AgNPs AgNPs ought ought to get more thoughtfulness regarding an efficient comprehension of how incorporating biosynthesized AgNP AgNPss in agrar agraria ian n appl applic icat atio ions ns cont contrib ribut utes es towa toward rd the the farm farmer er’s ’s bene benefit fits. s. One One impo importa rtant nt poin pointt is that that bio biosynt synthe hesi sizzed AgNP AgNPss have have demo demons nstr trat ated ed thei theirr wort worth h for for agri agricu cult ltur ural al appl applic icati ation onss by perf perfor ormi ming ng two two note notewo wort rthy hy undertakin undertakings, gs, viz., plant development development upgrade upgrade and plant malady management. Hence, now it is high time when future studies must be coordinated toward upgrading the utility of biosynthesized AgNPs in regular environments keeping in mind the end goal to predict their future agricultural extension.
2.10 2.10
Conc Conclu lusi sion on
In recent recent past, past, diseas diseasee manage managemen mentt strate strategie giess have have been been inclin inclined ed to much much safer safer altern alternativ atives es due to concerns over hazardous effects of chemical cal pest pestic icid ides es on huma human n and and plan plantt heal health th.. Biological control of plant pathogens employing livi livin ng micro icroor orga gani nissms offe offerrs such uch saf safe
2
Formulation Technology of Biocontrol Agents: Present Status tus and Future Prospects
alternative alternative to the chemicals. chemicals. In order to improve the efficiency and shelf life of biocontrol agents, various formulations based on solid and liquid carriers have been developed. Maximum care is required in biopesticide production and formulation as it contains live organisms. It is quite a confirmation that biopesticides will have a more extensive use and share greater market space in the future. Specific procedures and technologies have been recently developed that would significantly affect biopesticide formulations. Selection of abiotic stress tolerance or rhizospheric competence could allow a wider range of applications (Keswani 2015 (Keswani 2015). ). Moreover, fungicide tolerance screen screening ing would would promot promotee integr integrate ated d manage manage-ment with reduced chemical inputs. grateful to the Department Department of Acknowledgments HBS is grateful Biotechn Biotechnolog ology y (BT/PR (BT/PR5990 5990/AG /AGR/5/ R/5/587/ 587/2012 2012), ), New Delhi, for providing financial support. KB is grateful to BHU for financial assistance. BKS and CK are thankful to ICAR-A ICAR-AMAA MAAS S (Project (Project No. P27/131) P27/131) for financial financial support. support.
References Abbasi PA, Weselowski B (2015) Efficacy of Bacillus formulati ations ons,, copper copper hydrox hydroxide ide,, subtilis QST 713 formul and and their their tank tank mixes mixes on bact bacter erial ial spot spot of toma tomato. to. Crop Prot 74:70–76 Alam G (2000) A study of biopesticides and biofertilizers in Haryan Haryana, a, India, India, Gateke Gatekeepe eperr series series no. 93. IIED, IIED, London Arora NK, Khare E, Maheshwari DK (2010) Plant growth promoting rhizobacteria: constraints in biofor bioformu mula lati tion on,, comme commerc rcia ializ lizat atio ion, n, and and futur futuree strategies. In: Maheshwari DK (ed) Plant growth and heal health th prom promot oting ing bact bacter eria ia.. Sprin Springe ger, r, Berli Berlin, n, pp 97–116 Backman PA, Rodriguez–Kabana R (1975) A system for the growth and delivery of biological control agents to the soil. Phytopathology 65:819–821 Batta YA (2004) Postharvest biological control of apple gray mold by Trichode Trichoderma rma harzian harzianum um Rifai formulated in an invert emulsion. Crop Prot 23:19–26 Bisen K, Keswani C, Mishra S, Saxena A, Rakshit A, Sing Singh h HB (201 (2015) 5) Unre Unreal aliz ized ed pote potent ntia iall of seed seed bioprim biopriming ing for versat versatile ile agricu agricultu lture. re. In: Rakshi Rakshitt A, Singh HB, Sen A (eds) Nutrient use efficiency: from basics to advances. Springer, New Delhi, pp 193–206 Chio Chiou u AL, AL, Wu WS (200 (2003) 3) Form Formul ulat atio ion n of Bacillus B190 for for cont contro roll of lily lily grey grey amyloliquefaciens B190 mould ( Botrytis elliptica). J Phytopathol 151:13–18
49
Churchill BW (1982) Mass production of microorganisms for biological control. In: Charudattan R, Walker HL (eds (eds)) Biol Biolog ogic ical al cont contro roll of weed weedss with with plan plantt pathogens. Wiley, New York, pp 139–156 Colyer PD, Mount MS (1984) Bacterization of potatoes with Pseudomonas putida and its influence on post–harvest harvest soft rot diseases. diseases. Plant Dis 68:703–706 68:703–706 Commar Commaree RR, Nandakum Nandakumar ar R, Kandan Kandan A, Suresh Suresh S, Bharathi M, Raguchander T, Samiyappan R (2002) Pseudomonas fluorescens based bio–formulation for the manage managemen mentt of sheath sheath blight blight disease disease and leaf leaf folder insect in rice. Crop Prot 21:671–677 Connick WJ Jr (1988) Formulation of living biological control agents with alginate. In: Cross, Scher HB (eds) Pesticide formulations: innovations and developments, developments, vol 371, ACS symposium series. American Chemical Society, Washington, Washington, DC, pp 241–250 Connick WJ Jr, Lewis JA, Quimby PC (1990) Formulation of biocontrol agents for use in plant pathology. In: Baker RR, Dunn PE (eds) New directions in biological control: alternatives for suppressing agricultural pests and diseases. Alan R. Liss, New York, pp 345–372 Crozier J, Arroyo C, Morales H, Melnick RL, Strem MD, Vinyard BT, Bailey BA (2015) The influence of formulatio mulation n on Trichoderma biologi biological cal activi activity ty and fros frosty ty pod pod rot rot mana manage geme ment nt in Theobroma Theobroma cacao. Plant Pathol 64(6):1385–1395 Daigle DJ, Cotty PJ (1995) Formulating atoxigenic Aspergillus flavus for field release. Biocontrol Sci Technol 5:175–184 Dand Dandur uran and d LM, LM, Morr Morraa MJ, MJ, Chav Chaver erra ra MH, MH, Orse Orserr CS (1994) (1994) Surviv Survival al of Pseudomonas spp. spp. in air air drie dried d mineral powders. Soil Biol Biochem 26:1423–1430 DeLucca AJ, Connick WJ II, Fravel DR Jr (1990) Use of bacterial alginates to prepare biocontrol formulations. J Indust Microbiol 6:129–134 El–Say El–Sayed ed W (2005) (2005) Biolog Biologica icall control control of weeds weeds with with pathog pathogens ens:: curren currentt status status and future future trends. trends. J Plant Plant Dis Prot 112:209–221 Escande AR, Laich FS, Pedraza MV (2002) Field testing of honeybee‐dispersed Trichoderma spp. to manage sunflower head rot (Sclerotinia Sclerotinia sclerotiorum sclerotiorum). Plant Pathol 51:346–351 Fravel DR, Marois JJ, Lumsden RD, Connick WJ (1985) Encapsulation of potential biocontrol agents in alginate. Phytopathology 75:774–777 Garibaldi A, Brunatti F, Gullino ML (1987) Evaluation of several antagonists and different methods of application against fusarium wilt of carnation. OEPP/EPPO Bull 17:625–629 Ghor Ghorma made de V, Desh Deshpa pand ndee MV, MV, Pakn Paknik ikar ar KM (201 (2011) 1) Perspectives for nano–biotechnology enabled protectio tion and and nutr nutrit itio ion n of plan plants ts.. Biote iotech chno noll Adv Adv 29:792–803 Glare T, Caradus J, Gelernter W, Jackson T, Keyhani N, Ko¨ hl J, Stewart A (2012) Have biopesticides come of age. Trend Biotechnol 30:250–258 Harman GE (1991) Seed treatments for biological control of plant disease. disease. Crop Prot 10:166–171 10:166–171
50 Harman GE, Taylor AG (1988) Improved seedling performance by integration of biological control agents at favourable pH levels with solid matrix priming. Phytopathology 78:520–525 Hu X, Roberts DP, Xie L, Maul JE, Yu C, Li Y, Liao X (2014) Formulations of Bacillus Bacillus subtilis BY–2 suppress Sclerotinia sclerotiorum on oilseed rape in the field. Biol Control 70:54–64 Huber DM, El–Nasshar H, Moore LW (1989) Interaction between between a peat carrier carrier and bacteri bacterial al seed seed treatmen treatments ts evaluated for biological control of the take–all disease of wheat (Triticum aestivum L.). Biol Fert Soils 8:166–171 Jacobsen BJ, Backman A (1993) Biological and cultural plant disease controls: alternatives and supplements to chem chemic ical alss in IPM IPM syst system ems. s. Plan Plantt Dis Dis (USA (USA)) 77:311–315 Jager G, Velvis H (1985) Biological control of Rhizocto Rhizoctonia solani on potatoes by antagonists. 4. Inoculation of seed seed tubers tubers with with Verticillium other Verticillium biguttatum biguttatum and other antagonists in field experiments. Neth J Plant Pathol 91:49–63 Jayara Jayarajj J, Radhak Radhakrish rishnan nan NV, NV, Velazh Velazhaha ahan n R (2006) (2006) Deve Develo lopm pmen entt of form formul ulat atio ions ns of Trichoderma harzianum strain M1 for control of damping–off of tomato tomato caused caused by Pythium aphaniderma Arch aphanidermatum tum. Arch Phytopathol Plant Protec 39:1–8 Jo YK, Kim BH, Jung G (2009) Antifungal activity of silver silver ions ions and nanopa nanopartic rticles les on phytop phytopath athoge ogenic nic fungi. Plant Dis 93:1037–1043 Jones Jones KA, Burges Burges HD (1997) (1997) Produc Productt stabilit stability: y: from experimental experimental preparation preparation to commercial commercial reality. In: Evans Evans HF (ed) (ed) Microb Microbial ial insectic insecticide ides: s: novelty novelty or necess necessity ity?? Sympos Symposium ium procee proceedin dings gs no. 68. British British Crop Protection Council, Farnham, pp 163–171 Jones RW, Pettit RE, Taber RA (1984) Lignite and silage: carrier and substitute for application of fungal biocontrol agents to soil. Phytopathology 74:1167–1170 Keinath AP, Fravel DR, Papavizas GC (1990) Evaluation of formulations of Talaromyces flavus for biocontrol of Verticillium Verticillium wilt of potato. Biol CultTests 7:27 Keswan Keswanii C (2015) (2015) Ecofrie Ecofriendl ndly y manage managemen mentt of plant plant diseases by biosynthesized secondary metabolites of doi:10.5281/zenodo. Trichoderma spp. J Brief Idea. doi:10.5281/zenodo. 15571 Kesw Keswan anii C, Sing Singh h SP SP,, Sing Singh h HB (201 (2013a 3a)) Beauveria status, mode mode of action action,, applica applicatio tions ns and bassiana: status, safety issues. Biotech Today 3:16–20 Keswani C, Singh SP, Singh HB (2013b) A superstar in biocontrol biocontrol enterprise: enterprise: Trichoderma spp. spp. Biotec Biotech h Today 3:27–30 Keswani C, Mishra S, Sarma BK, Singh SP, Singh HB (2014) Unraveling the efficient application of secondary ary meta metabo boli lite tess of vari variou ouss Trichoderma. Appl Microbiol Biotechnol 98:533–544 Khater Khater HF (2012) Prospects of botanical botanical biopesticides biopesticides in insect pest management. J Appl Pharm Sci 2:244–259 Kim SW, Jung JH, Lamsal K, Kim YS, Min JS, Lee YS (2012) (2012) Antifu Antifunga ngall effects effects of silver silver nanopa nanopartic rticles les
C. Keswani et al. (AgNPs (AgNPs)) agains againstt variou variouss plant plant pathog pathogeni enicc fungi. fungi. Mycobiology 40:53–58 Kloepp Kloepper er JW, Schrot Schroth h MN (1981) (1981) Develo Developme pment nt of a powder formulation of rhizobacteria for inoculation of potato seed pieces. Phytopathology 71:590–592 Knowles Knowles A (2005) New developments developments in crop protection protection product formulation, Agrow reports. T and F Informa UK Ltd, London, pp 153–156 Know Knowle less A (200 (2006) 6) Adju Adjuva vant ntss and and addi additiv tives es,, Agro Agrow w reports. T&F Informa UK Ltd, London, pp 126–129 Lee JP, Lee SW, Kim CS, Son JH, Song JH, Lee KY, Moon BJ (2006) Evaluation of formulations of Bacil Bacillus licheniformis for the biological control of tomato gray mold caused by Botrytis cinerea. Biol Control 37:329–337 Lewis JA, Lumsd msden RD (20 (2001) Biocontrol of damping–off of greenhouse–grown crops caused by Rhizoctonia solani with a formulation of Trichoderma Trichoderma spp. Crop Prot 20:49–56 Lewis JA, Papavizas GC (2007) Characteristics of alginate nate pelle pellets ts form formul ulat ated ed with with Trichoderma and Gliocladium and their effect on the proliferation of the fungi in soil. Plant Pathol 34:571–577 Lim Lim TK, TK, Rohr Rohrba bach ch KG (198 (1980) 0) Role Role of Penicillium strains in the development of pineapple funiculosum fruit diseases. Phytopathology 70:663–665 Lisansky SG (1985) Production and commercialization of pathogens. In: Scopes N, Hussey NW (eds) Biological pest control. Blanford Press, Poole, pp 210–218 Mahaffee WF, Backman PA (1993) Effect of seed factors on spermosphere and rhizosphere colonization of cotton by Bacillu GB03. Phytopatholo Phytopathology gy Bacilluss subtili subtiliss GB03. 83:1120–1125 Manjul Manjulaa K, Podile Podile AR (2001) (2001) Chitin– Chitin–sup supple plemen mented ted formula formulation tionss improve improve biocon biocontrol trol and plant plant growth growth promoting efficiency of Bacillus subtilis AF 1. Can J Microbiol 47:618–625 Mathivanan N, Prabavathy VR, Vijayanandraj VR (2005) Applica Application tion of talc talc formula formulation tionss of Pseudomonas fluorescens Migula and Trichoderma viride Pers. ex SF Gray Gray decr decrea ease se the the shea sheath th blig blight ht disea disease se and and enha enhanc ncee the the plan lant grow growth th and yiel yield d in ric rice. J Phytopathol 153:697–701 Mbarga Mbarga JB, Begoud Begoudee BAD, BAD, Ambang Ambang Z, Meboma Meboma M, Kuate J, Schiffers B, Ten Hoopen GM (2014) A new oil–based formulation of Trichoderma Trichoderma asperellum for the biolog biologica icall contro controll of cacao cacao black black pod diseas diseasee caused caused by Phytophthora Biol Cont Contro roll Phytophthora megakarya megakarya. Biol 77:15–22 McLean KL, Swaminathan J, Frampton CM, Hunt JS, Ridgway HJ, Stewart A (2005) Effect of formulation on the rhizosphere competence competence and biocontrol ability of Trichode C52. Plan Plantt Path Pathol ol Trichoderma rma atrovi atrovirid ridee C52 54:212–218 Merriman Merriman PR, Price RD, Kollmorgen JF (1974) Effect Effect of seed inoculation with Bacillus subtilis and Streptomyces griseus on the growth of cereals and carrots. Aust J Agric Res 25:219–226
2
Formulation Technology of Biocontrol Agents: Present Status tus and Future Prospects
Mish Mishra ra S, Sing Singh h HB (201 (2015) 5) Bios Biosyn ynth thes esiz ized ed silv silver er nanopart nanoparticle icless as a nanowe nanoweapo apon n against against phytopat phytopathoge hogens: ns: exploring their scope and potential in agriculture. App Microbiol Biotechnol 99:1097–1107 Mishra S, Singh BR, Singh A, Keswani C, Naqvi AH, Singh HB (2014) Biofabricated silver nanoparticles act as a strong fungicide against Bipolaris sorokiniana caus causin ing g spot spot blot blotch ch dise diseas asee in whea wheat. t. PLoS PLoS One One 9:97881 Mishra S, Singh A, Keswani C, Saxena A, Sarma BK, Singh HB (2015) Harnessing plant–microbe interaction for enhanced enhanced protection protection against against phytopathog phytopathogens. ens. In: Arora NK (ed) Plant microbes symbiosis: applied facets. Springer, New Delhi, pp 111–125 NAAS (2013) Biopesticides – quality assurance. Policy paper paper No. 62. Nation National al Academ Academy y of Agricu Agricultu ltural ral Sciences, New Delhi, p 20 Nagtzaam MPM, Bollen GJ (1994) Long shelf life of coating g materi material al of pellet pelleted ed Talaromyces Talaromyces flavus in coatin seed. Eur J Plant Pathol 100:279–282 Nelson EB (1988) Biological control of Pythium Pythium seed rot and and pre pre emer emergen gence ce damp dampin ing– g–of offf of cotto cotton n with with Enterobacter cloacae and Erwinia herbicola applied as seed treatments. Phytopathology 72:140–142 Park HJ, Kim SH, Kim SJ, Choi SH (2006) A new composition position of nanosized silica–silver silica–silver for control control of various plant diseases. diseases. Plant Pathol Pathol J 22:295–302 22:295–302 Parke JL, Rand RE, Joy AE, King EB (1991) Biological control of Pythium damping–off and Aphanomyces root Pythium damping–off rot of peas by application of Pseudomonas cepacia or P. fluorescens to seed. Plant Dis 75:987–992 Patanjali PK, Raza SK (2013) Recent advances in pesticide formulation technology. In: Abstract of international conference on emerging issues on health effects of pesticides residues in food and environment– ultimate challenges and research opportunities, Food and Drug Drug Toxico Toxicolog logica icall Resear Research ch Institu Institute, te, Hydrab Hydrabad, ad, 12–13 December 2013 Paulitz Paulitz TC, Anas O, Fernan Fernando do DG (1992) (1992) Biolog Biologica icall control control of Pythium damping–of damping–offf by seed treatment treatment with Pseudomonas putida: relationship with ethanol production by pea and soybean seeds. Biocontrol Sci Technol 2:193–201 Peeran Peeran FM, Krishn Krishnan an N, Thanga Thangaman manii PR, Gandhi Gandhi K, Thiruv Thiruveng engada adam m R, Kuppus Kuppusamy amy P (2014) (2014) Develo Developpment and evaluation of water–in–oil formulation of (FP7) against Pseudomonas fluorescens (F Colletotrichum musae incident of anthracnose disease in banana. Eur J Plant Pathol 138:167–180 Pusey PL, Wilson CL (1984) Postharvest Postharvest biological biological control of stone fruit brown rot by Bacillus subtilis. Plant Dis 68:753–756 Rabindran Rabindran R, Vidhyasekar Vidhyasekaran an P (1996) Development Development of a formulation of Pseudomonas fluorescens PFALR2 for mana manage geme ment nt of rice rice shea sheath th blig blight ht.. Crop Crop Prot Prot 15:15–721 Rao GVR, Rupela OP, Rao VR, Reddy YVR (2007) Role of biopesticides biopesticides in crop protection: protection: present status and future prospects. Ind J Plant Prot 35:1–9
51
Ristaino Ristaino JB, Lewis JA, Lumsden RD (1994) (1994) Influence Influence of Gliocladium virens and delivery systems on biological control of southern blight on carrot and tomato in the field. Plant Dis 78:153–156 Singh HB (2006) Trichoderma: a boon for biopesticides industry. industry. J Mycol Plant Pathol 36:373–384 36:373–384 Singh HB (2012) Regulatory requirements, quality control, trol, commer commercia cializ lizati ation on and IPR relate related d issues issues by bio– bio–pe pest stic icid ides es.. In: In: Sing Singh h HP, HP, Chow Chowda dapp ppaa P, Chakraborty, Podile AR (eds) Molecular approaches for plant fungal disease management. Westville, New Delhi, pp 228–224 Singh HB, Singh A, Nautiyal Nautiyal CS (2003) CommercializaCommercialization of biocontrol biocontrol agents: problems and prospects. In: Rao Rao GP, GP, Mano Manoha hach char arii C, Bhat Bhat DJ, DJ, Raja Rajak k RC RC,, Lakhnapal TN (eds) Frontiers of fungal diversity in India. India. Interna Internation tional al Book Book Distrib Distributi uting ng Compan Company, y, Lucknow, pp 847–861 Sing Singh h HB, HB, Sing Singh h A, Sing Singh h SP SP,, Naut Nautiy iyal al CS (200 (2004a 4a)) Commercialization of biocontrol agents: the necessity and its impact on agriculture. In: Singh SP, Singh HB (eds (eds)) Eco– Eco–ag agri ricu cult ltur uree with with bioa bioaug ugme menta ntatio tion: n: an emerging concept. Rohitashwa printers, Lucknow, pp 1–20 Singh HB, Singh S, Singh A, Nautiyal CS (2004b) Mass production, formulation and delivery system of fungal and bacter bacterial ial antago antagonis nistic tic organi organisms sms in India. India. In: Sing Singh h SP SP,, Sing Singh h HB (eds (eds)) Eco– Eco–ag agric ricult ultur uree with with bioaugmenta bioaugmentation: tion: an emerging emerging concept. concept. Rohitashwa Rohitashwa printers, Lucknow, pp 53–69 Singh HB, Ahmad A, Srivastav S, Nautiyal CS (2006) A synergistic composition useful as bioinoculant (PCT No. WO 2007/11068A2) Sing Singh h HB, HB, Sing Singh h BN, BN, Sing Singh h SP SP,, Sarm Sarmaa BK (201 (2012) 2) Explori Exploring ng differ different ent avenue avenuess of Trichoderma as a potent potent bio–fu bio–fungi ngicid cidal al and plant plant growth growth promot promoting ing candidate–an overview. Rev Plant Pathol 5:315–426 Singh HB, Singh A, Sarma BK, Upadhyay DN (2014) Trichoderma Trichoderma viride 2% WP (Strain No. BHU–2953) formulation suppresses tomato wilt caused by Fusarf. sp. lycopersici and and chil chilli li ium ium oxys oxyspo poru rum m f. dampin damping–o g–off ff caused caused by Pythium aphanidermat aphanidermatum um effectively effectively under different different agroclimatic agroclimatic conditions. conditions. Int J Agri Environ Biotechnol 7:313–320 Sivakumar G, Sharma RC, Rai SN (2000) Biocontrol of banded leaf and sheath blight of maize by peat based formulati ation on.. India Indian n Pseudomonas fluorescens formul Phytopathol 53:190–192 Stack JP, Kenerley CM, Pettit RE (1987) Influence of carbon carbon and nitroge nitrogen n source sources, s, relative relative carbon carbon,, and nitrog nitrogen en concen concentra tratio tions ns and soil soil moistu moisture re on the grow growth th in nons nonste teri rile le soil soil of soil soilbo born rnee fung fungal al antagonists. Can J Microbiol 33:626–631 Stack JP, Kenerley CM, Pettit RE (1988) Application of biocon biocontro troll agents, agents, in biocon biocontro troll of plant plant diseas diseases. es. CRCPress, Boca Raton, pp 43–54 Stowell Stowell LJ (1991) Submerged Submerged fermentation fermentation of biological biological herbicides. In: TeBeest D (ed) Microbial control of weeds. Chapman and Hall, New York, pp 225–261
52 Suslow Suslow T, Schrot Schroth h MN (1982) (1982) Rhizob Rhizobact acteria eria of sugar sugar beets: effects of seed application and root colonization on yield. Phytopathology 72:199–206 Tadros Tadros F (2005) (2005) Applie Applied d surfact surfactant ants, s, princi principle pless and appl applic icat atio ions ns.. Wile Wiley– y–VC VCH H Verl Verlag ag GmbH GmbH and and Co. KGA, Weinheim, pp 187–256 Taylor AG, Min TG, Harman GE, Jin X (1991) Liquid coating formulation for the application of biological seed treatments of Trichoderma Trichoderma harzianum. Biol Control 1:16–22 Thanga Thangavel velu u R, Palani Palaniswa swami mi A, Velazh Velazhaha ahan n R (2004) (2004) Mass production of Trichoderma Trichoderma harzianum for managing aging fusari fusarium um wilt wilt of banana banana.. Agri Agri Eco Environ Environ 103:259–263
C. Keswani et al. Tomlin C (ed) (1994) The pesticide manual. British Crop Protec Protectio tion n Counci Council/R l/Roya oyall Societ Society y of Chemist Chemistry, ry, Farnham Tronsmo A, Dennis C (1983) The use of Trichoderma species to control strawberry fruit rots. Neth J Plant Pathol 83:449–455 Vidhyasekaran P, Muthamilan M (1995) Development of formulations of Pseudomonas fluorescens for control of chickpea wilt. Plant Dis 79:782–786 79:782–786 Weller DM, Cook RJ (1983) Suppression of take–all of wheat by seed trea reatmen ments with ith fluorescent Pseudomonads. Phytopathology 73:463–469 Wilson Wilson CL, Pusey Pusey PL (1985) (1985) Potent Potential ial for biolog biologica icall control of post harvest diseases. Plant Dis 69:375–378