REVIEW OF LITERATURE (Garret, 1965). (Cook and Baker, 1983) BIOCONTROL AGENTS

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1 REVIEW OF LITERATURE Integrated Pest Management or IPM, is a method used to control pests in an environmentally responsible manner. By reducing our dependence on pesticides, IPM protects the environment and our health. It also saves money. IPM can be applied wherever pests are found: on and in farms, schools, homes, hospitals, restaurants, golf courses and home gardens. IPM combines different techniques to prevent pest damage without harming the environment. Pests can include insects and mites, rodents and certain birds, plant diseases and weeds. IPM practices include monitoring, modifying pest habitat, protecting natural enemies and when needed, the use of pesticides. Biological control can be defined as- any condition under which, or practice whereby, survival and activity of a pathogen is reduced through the agency of any other living organism (except man himself) wit the result that there is a reduction in the incidence of disease caused by the pathogen (Garret, 1965). Broadly speaking, it is a principle of cultural control of plant pathogens that principally involves alteration of biotic and abiotic environments from one favours disease/pathogen to one that discourages accumulation of infective or parasitic material and reduces the activity of the pathogen. Biological control was defined by Baker and Cook (1974) as the reduction of inoculum density or disease producing activities of a pathogen of parasite in its active or dormant state, by one or more organisms, accomplished naturally or through manipulation of the environment, host, or antagonists, or by mass introduction of one or more antagonists. Subsequently, they (Cook and Baker, 1983) revised the definition to biological control is the reduction of the amount of inoculum of disease producing activity of pathogen accomplished by one or more organisms other than man. BIOCONTROL AGENTS Biocontrol agents are microorganisms that adversely affect the population of another (e.g. target pathogen) growing in association with them. Generally biocontrol agents have potential to interfere in the life process of plant pathogens. Biocontrol agents include virtually all classes of organisms e.g.- fungi, bacteria, nematodes, protozoa, viruses and seed plants. 24

2 Fungi are by far the most extensively researched group of biocontrol agents and they have been used against aerial, root and soil microbes. These potential biocontrol fungi are mostly saprophytic in nature and proliferate abundantly in various natural soils. Interest in their use for control of aerial plant pathogens has developed more slowly than in the case of root pathogen and this probably reflects the relative dearth of information on the ecology of microorganisms on the aerial surfaces in comparison with those in the soil (Mukhopadhyay and Mukherjee, 1998). The important genera of fungi used as biocontrol agents against plant pathogens are Trichoderma, Gliocladium, Aspergillus, Penicillium Neurospora, Chaetomium, Dactylella, Arthrobotrys, Catenaria, Paecilomyces, Glomus, etc. The use of bacterial flora for management of plant diseases and yield improvement started in the early part of 20 th century. A number of bacterial strains have so far been tried as biocontrol agents. These are Agrobacterium, Actioplanes, Alcaligenes, Arthrobacter, Azotobacter, Bacillus, Enterobacter, Erwinia, Flavobacterium, Pseudomonas, Pasteuria, Rhizobium, Serratia, Streptomyces, etc. The most widely researched bacterium as a biocontrol agent is Pseudomonas fluorescens. Integrated Pest Management Raman et al. (2003) discussed the applications of biotechnology in integrated pest management, specifically covering the following: induction of resistance in crops through genetic transformation (with emphasis on Bt [Bacillus thuringiensis] transgenic plants); agronomic benefits of transgenic crops (rice, maize, soyabean and potato) in developing countries; main approaches for the production of new biological control agents using genetic engineering; new diagnostic tools based on monoclonal antibodies and molecular markers; DNA markers, mapping, and application for developing resistant plants; policy issues affecting access to new biotechnologies; pest resistance management; regulatory policy options for resistance management strategy (licensing and labelling); use of IPM to delay pest resistance; public-private linkage considerations and future prospects for biotechnology in IPM ZhenYing et al. (2003) studied the development of IPM (from 1950 to the early 1970s, from 1974 to 1982, and from 1983 to the present), IPM organizational structure, IPM research and practice (pest monitoring and forecasting, cultural control, biological control 25

3 and pest resistance) and examples of successful IPM application (rice, wheat and maize) in China. Santha-Govind and Perumal, (2004) stated that the production of rice mainly depends upon the management of pests and diseases, which takes a major share in the total production cost. Integrated pest management (IPM) is the eco-friendly solution to overcome the pest management problems. A study was undertaken to identify the constraints faced by the farmers in the adoption of IPM practices and to suggest appropriate corrective measures to make farming a viable and profitable enterprise. The major findings on the constraints faced in the adoption of IPM technologies among IPM and Non IPM farmers are analysed. Krishnamurthy et al. (2005) studied in five districts (Mandya, Mysore, Shimoga, Hassan and Bangalore Rural) of Karnataka, India, the impact of farmers' field schools (FFSs) on knowledge and measured the attitude of farmers and extension personnel in respect to IPM practices in rice farming. The respondents consisted of 60 trained and 60 untrained rice farmers and 60 trained and 60 untrained extension personnel. Results revealed that 53% of the trained farmers and 48% of the trained extension personnel had a high level knowledge on IPM practices in rice cultivation. The study also revealed that 43% of the trained farmers and 37% of the trained extension personnel had a more favourable attitude towards IPM practices in rice cultivation. Further, it also revealed that 36% of the farmers and 42% of the extension personnel had a low level of knowledge regarding IPM practices for rice, whereas 33% of the farmers and 46% of the extension personnel had a less favourable attitude towards IPM in rice cultivation. Patel et al. (2006) tested Isolates of Beauveria bassiana (Balsamo) Vuillemin for biological control of rice stink bug, Oebalus pugnax (F.) in the laboratory, in small-plot field experiments compared with conventional insecticides, and in a large-plot experiment to determine the spread and persistence of the fungus. The overall impact of B. bassiana was moderate on O. pugnax nymphs and minimal on adults in the small-plot experiments. A single application of B. bassiana reduced rice stink bug nymphs on six of nine sampling dates and adults on two of nine sampling dates from 2-10 d after application in the three small-plot experiments, and prevalence of the fungus was higher in the B. bassiana treatment than in controls for nymphs on four dates versus none for 26

4 adults. A single application of chemical insecticide reduced total rice stink bug numbers more than B. bassiana for at least 7 d in small-plot experiments, whereas a double application was more effective than B. bassiana for 10 d against nymphs. Beauveria bassiana was nearly as effective as a single application of chemical insecticide in suppressing rice stink bug numbers 7-8 d after application. Mixtures of B. bassiana and chemical insecticide provided better control of rice stink bug than a single application of either material alone. Thus, B. bassiana has potential for integrated management programs of O. pugnax in rice, because it was moderately effective against nymphs and had an additive effect with insecticides. Peshin and Kalra (2006) selected a sample of 150 farmers (135 of which were trained in integrated pest management or IPM and 15 were untrained) in Ludhiana district, Punjab, India to study the extent and level of adoption of IPM practices in rice crop. There was not much difference in the adoption of different cultural practices between IPM and non-ipm farmers, but the IPM farmers had adopted resistant cultivars in a large scale. IPM farmers had applied lesser number of pesticide sprays (1.36 per season) compared to non-ipm farmers (2.47 per season). Ten percent of the IPM-trained farmers did not apply any insecticide. To have desired impact of IPM, the employment of farmers is necessary, which was not the case in all villages. Sithanantham, et al. (2007) added that Thrips are an important group of sucking insects causing substantial yield losses in several tropical crops, as direct pests and/or as virus vectors. The emerging interest at national level in organic farming and export agriculture calls for development of appropriate and wide range of eco-friendly and biological products for thrips management. Entomopathogens (like Verticillium lecani) have shown promise for augmentative biological control of Scirtothrips dorsalis, and there was scope for identifying more adapted and virulent strains of the entomopathogens, There was need to focus on research and development (R&D) with public-private partnerships, for widening the range and refining the technology options as well as the integration of different biological control agents with other pest control technologies, besides linking them to the crop management practices so as to evolve holistic Integrated Pest Management (IPM) strategies. 27

5 Kaur et al. (2007) demonstrated bio-intensive pest management of leaf folder and stem borer on basmati rice at village Karni Khera (District Ferozepur), Punjab over 60 hectares during Bio-intensive management package included one application of cartap hydrochloride, 25 kg/ha) and seven weekly releases of Trichogramma chilonis and T /ha each starting from 30 days after transplantation. The mean per cent leaves folded (2.02), dead hearts (3.05), white ears (5.45) and yield (43.99 q/ha) in BPIM were on par with chemical control, (mean per cent leaves folded (1.77), dead hearts (2.62), white ears (4.48) and yield (44.83 q/ha)), and both the treatments were significantly better than control. The cost: benefit ratio was 1:4.01 and 1:4.68 for biointensive management practice and chemical control, respectively. Alka et al. (2007) examined pesticide use pattern and adoption of integrated pest management (IPM) practices in rice, vegetables and cotton. Estimated values of farmers' willingness to pay for pesticide hazard reduction indicated that a marker exists for environmentally friendly pesticides in the study areas. Alka et al. (2008) studied adoption of Integrated Pest Management (IPM) practices on paddy in Haryana and assessed the impact of key socio-economic and institutional factors on adoption. A survey was conducted in on 83 farmers cultivating paddy in the Karnal and Kaithal districts of Haryana. The study shows that technology awareness through formal crop-specific IPM training provided by farmers' field schools was extremely important for wider adoption of IPM in the study area. The study has found mixed evidence about the relationship between farm-size and adoption of IPM practices. In the case of paddy, a negative relationship was observed. Chen et al. (2008) studied efficacy and economic benefits of 4 rice pest control methods in Zhejiang, China. Damage in rice fields due to the insect pests was severe, and the economic benefit decreased when pesticide was not sprayed during the rice growth period. Standard spray and farmers' spray exhibited similar control efficacy against planthoppers (Nilaparvata lugens, Sogatella furcifera and Laodelphax striatellus) and leaf roller (Cnaphalocrocis medinalis). Grain agronomic character, rice yield and economic benefit were also similar for these treatments. However, the incidence of wilt sheath shoot rate caused by the striped stem borer (Chilo suppressalis) was highest for farmers' spray. The biological control system based on duck-rice interaction + 28

6 biopesticide spray was effective against insect pests, but this treatment resulted in lower rice yield than the standard spray (55.15% of the yield for this treatment) and farmers's spray (59.60%). However, higher economic return was obtained with the biological control system (by ~ yuan/667 m2) than with standard spray and farmers' spray. Lal et al. (2008) conducted an experiment in Bhagalpur, Bihar, India, to determine the constraints in adoption of IPM in rice cultivation. The overall percentage regarding the constraints pertaining to technology was 67.80, constraints pertaining to extension was and constraints pertaining to service, supply and marketing was and constraints pertaining to transfer of technology was in the adoption of IPM in rice cultivation. Ramandeep et al. (2008) conducted experiment on the management of rice leaf folder and stem borer in two rice varieties, viz., PR 116 and Basmati 386, in the organic field by comparing three treatments, viz. organic, recommended and integrated practices. Organic practices and integrated practices (seven releases of Trichogramma chilonis and T each at weekly interval starting 30 DAT) proved to be effective in the management of rice leaf folder and stem borer in both the rice varieties. Mariyono, (2008) studied Integrated pest management (IPM) technology disseminated since 1989 in Indonesia to cut down pesticide use, but the adoption and diffusion of the technology were still debated. This study aimed to estimate the models of demand for pesticides and to analyse the impact of IPM technology on pesticide use. There was an indication that IPM technology has been adopted by farmers. This is evidence that the IPM programme in Indonesia was successful in this area. Tripathi et al. (2008) stated that integrated pest management (IPM) is the integrated use of pest control strategies in a way that not only reduces pest population to satisfactory levels but is sustainable and non-polluting. IPM in India was as receiving high priority since about three decades, under the ICAR set up, the AICRP on IPM are in operation in collaboration with SAU. The goal of the IPM development is not just to maximize yields but also to increase farmer's income and welfare. Therefore, the socio-economy and culture of the local farmers need to be studied to determine proper strategy for extension and training of IPM technology. Farmers play an important role in the development and implementation of IPM. A project initiated in the Philippines based on the principle of 29

7 IPM is expected to help farmers increase vegetable yields while decreasing the use of potentially harmful pesticides. The project, was based on a successful small scale experiment carried out with farmers in the Philippines, where vegetable yields rose and total production costs fell by 20%, as farmers reduced expenditure on chemicals. Government of India launched IPM programme with FFS approach in 1992 on small scale for rice and cotton crops as the pesticides used for these crops contributes to more than 75% of total consumption. This introduction of IPM programme was well timed as chemical control against some pests such as Heliothis and red-hairy caterpillar (Amsacta sp)] had become ineffective and there was a desperate search for other alternatives. Among all these constraints, unavailability of location specific IPM package was a major constraint. This had to be developed by extension functionaries by involving farmers and scientists. In recent years farmers, non-government organizations, scientists and governmental and international institutions are engaging in joint efforts to replace chemical-intensive farming methods with alternative agro-ecological approaches. Singh et al. (2008) conducted a field experiment in Meerut, Uttar Pradesh, India, during the kharif season of 2005 to study the effects of cartap hydrochloride 4 G at 0.75 kg a.i./ha, Beauveria bassiana at 2.5 kg/ha, Bacillus thuringiensis at 1.5 kg/ha, imidacloprid 17.8 SL at 0.05%, Trichogramma japonicum at eggs/ha, Neem Gold at 5 ml/litre and Neem Gold at 3 ml/litre + T. japonicum at eggs/ha on yellow stem borer (YSB; Scirpophaga incertulas) and rice leaf folder (Cnaphalocrocis medinalis) infesting rice (cv. Basmati), and their predators (spiders). The biological pesticides were superior over the synthetic insecticides in conserving the spider population. The release of T. japonicum with botanicals reduced the populations of both pests and conserved the spider population. The cost: benefit ratio was highest with imidacloprid 17.8 SL (1:10.13) and lowest with Bacillus thuringiensis (1:1.90). Sharifi et al. (2008) developed Integrated pest management (IPM) as one of sustainable agricultural development components, based on synergy of empowering farmers, environmental friendly technology and integrated farm management. Effective IPM extension needs identification and analysis of related principal components with aim to policy making and planning by farmers' participation. The statistical population included 1145 of rice farmers. A sample of 90 farmers was selected by the use of 30

8 proportional random sampling method. The result of factor analysis showed that three components were extracted of IPM practices. The first factor called the optimal cultivation practices that explained 26% of the total variance and others were biological practices and physical-mechanical practices. Mohapatra et al. (2009) adopted Indigenous pest control practices by tribes at 7 locations in Tamil Nadu, India and studied the efficacy of the most popular practices (including the application of plant extracts and organic amendments) which were evaluated against pests of rice, groundnut, pulses, vegetables and stored grains under field and laboratory conditions. These practices were effective against the pests. Patel et al. (2009) conducted a field experiment during the kharif season from 2000 to 2004 in Nawagam, Gujarat, India, and evaluated the efficacy of integrated pest management (IPM) against C. medinalis on rice. Pooled data indicated that IPM exhibited a lower incidence of C. medinalis (3.60%) than farmers' practice (7.0%). Spider population ranged from to spiders/m2 for IPM, and from 2.30 to 9.50 spiders/m2 for farmers' practice. IPM also resulted in higher grain yield (4845 kg/ha), net return ( rupees/ha) and cost benefit ratio (1:1.14) than farmers' practice (3845 kg/ha, 9486 rupees/ha and 1:0.69, respectively). Raikar et al. (2009) conducted field experiment in Mugad, Karnataka, India, during the kharif of 2005 and 2006 to study the effects of nutrient The application of 50% RDN through farmyard manure + 50:50:50 kg NPK/ha combined with the integrated pest management system resulted in the highest seed yield (3806 kg/ha). Insect Management Garg and Baranwal (1998) used an improved Basmati rice variety Taraori, a neembased pesticide, the parasitoid, Trichogramma japonicum and insecticides were used against leaf folder (Cnaphalocrocis medinalis) and Yellow Stem Borer (Scirpophaga incertulas), in an IPM approach during the kharif seasons in Haryana, India. The results revealed that IPM technology was as effective as the pesticidal approach in controlling pests and increasing rice yields. Furthermore, it was environmentally safe and economically viable. Chai et al. (2000) reported the pathogenicity of 37 isolates of Metarhizium anisopliae to Lissorhoptrus oryzophilus and the sensitivity of L. oryzophilus at different stages to M. 31

9 anisopliae were tested. The results showed that 6 isolates of M. anisopliae parasitized over 90% of L. oryzophilus. L. oryzophilus was more sensitive to the infection of M. anisopliae at the reproductive stage, and intermediate at the overwintering stage and less sensitive at the emerging stage. Chen et al. (2000) studied Metarhizium anisopliae to control rice water weevil, Lissorhoptrus oryzophilus at the pre-oviposition stage in the field [in China]. Results showed that the adult population of L. oryzophilus was reduced by 92.5% 13 d after spraying at a rate of 1014 spores/ha. Larval and adult populations in the coming generation decreased to 2.12 and 0.30 per hill, while those in the check the populations were 8.40 and 4.17 per hill, respectively. El-Khayat (2000) evaluated the effects of some plant extracts (Thuja orientalis leaves and fruits and Melia azedarach fruits extracted with petroleum ether) and also of two bioinsecticides (Dipel [Bacillus thuringiensis subsp. kurstaki] and Biofly [Beauveria bassiana]) at two concentrations (5, 10 mg/50 g) in the laboratory against three stored grain pests, Sitophilus oryzae and Rhyzopertha dominica adults and Trogoderma granarium larvae. The results indicated that Thuja extracts of leaves and fruits were more effective against the three insects causing 80.6, 68.2 and 53.3%, and 56.2% mortality to S. oryzae adults R. dominica adults and T. granarium larvae, respectively, after 3 weeks of treatments. The treatment with Dipel and Biofly at the higher concentration (10 mg/50 g grains) gave moderate mortality rates of the three insects. These biocontrol agents showed more effect on S. oryzae than on R. dominica and/or T. granarium. The plant extracts gave good protection to wheat grains against the three insect pests during the period of storage (3 months). The bioinsecticide (Dipel) gave accepted degree of protection to wheat grains during the 3 months of storage. Legaspi et al. (2000) evaluated Beauveria bassiana (strain GHA) against the Mexican Rice Borer, Eoreuma loftini, which causes damage in south Texas estimated between $10 and $20 million annually. They performed bioassays against the sugarcane borer, Diatraea saccharalis, the key pest in other sugarcane growing areas. In the bioassays, E. loftini was substantially more susceptible to B. bassiana than D. saccharalis, based on both 5-day LD50 values and survival times. A commercial oil-based formulation of B. bassiana was evaluated in the field using the following treatments: oil alone (control), B. 32

10 bassiana + oil, and B. bassiana + Silwet L-77 carrier at an application rat of 5 x 1013 spores per hectare. Neither numbers of E. loftini per stalk, nor stalk damage (~20% bored internodes) were significantly affected by treatment. The application of B. bassiana + Silwet significantly affected the numbers of internodes showing high damage, but not those with low or medium damage. Analysis of yield data and juice quality showed to significant treatment effects. Carballo et al. (2001) studied the various B. bassiana isolates against control of pepper weevil (Anthonomus eugenii). Several concentrations of the fungus in water and oil suspension were evaluated to determine the half-lethal concentration (LC 50) using immersion and aspiration application methods. All isolates were pathogenic to the weevil, with the greatest % mortality, lowest lethal time (LT 50) and greatest yield of conidia on rice determined for the isolates 447, RL9-1, 113, 9205, 9218, 9006, 35 and 290. Dainingstar (2001) evaluated the efficacy of aqueous and methanolic leaf extracts from Ageratum conyzoides, Artemisia nilagirica, Eupatorium riparium and Lantana camara (50, 100, and 200 mg/ml) to inhibit the feeding activity of O. hyla hyla. Rice leaves were dipped into the extracts before being fed to adult grasshoppers for 6 days. Aqueous extracts were less effective than methanolic extracts in inhibiting the feeding activity of O. hyla hyla. Variation in the inhibitory action of the aqueous extracts at various concentrations was not significant. The aqueous extract of E. riparium showed the greatest feeding inhibition. Unlike the aqueous extracts, all methanolic extracts inhibited feeding completely with the increase in concentration and treatment duration. The methanolic extracts of A. conyzoides and E. riparium were most potent. Dal Bello et al. (2001) tested the virulence of ten different fungal isolates of: Beauveria bassiana, Metarhizium anisopliae, Verticillium lecanii and Paecilomyces farinosus to the rice weevil Sitophilus oryzae. A fungal mix of the most efficient isolates, B. bassiana ARSEF M. anisopliae ARSEF 2974, which caused the highest mortality, was assayed in combination with fenitrothion at a concentration lower (3 ppm) than the normal 6 ppm. Fungal inoculation of insects was conducted by spraying conidial suspensions of each fungus on wheat. Insecticide formulations were added by spraying wheat. Treated and untreated insects were incubated on durum wheat. Insects were kept 33

11 in a climatized chamber for 30 days and observations were module at 7, 14 and 30 days to record insect mortality. Highly significant differences were demonstrated for B. bassiana 5500 and 5501 and for M. anisopliae The level of mortality produced by treatments was: 6 ppm insecticide = 97.50%, B. bassiana ARSEF M. anisopliae ARSEF ppm insecticide = 74.17%, B. bassiana ARSEF M. anisopliae ARSEF 2974 = 50% and 3 ppm insecticide = 37.50%. There was a statistically significant difference (p < 0.05) among treatments. Govindan et al. (2001) tested the entomopathogenic fungus, B. bassiana was against the rice weevil S. oryzae. Bioassays were conducted by introducing 25 adult insects on 50 g of B. bassiana-mixed rice in glass jars maintained at 28 + or - 2 degrees C and 70% RH. Mortality was monitored at 5-day intervals until 25 days. At higher rates (7.6 log conidia/ml), B. bassiana produced a higher percentage of mortality up to 75.8% and F 1 adult emergence was reduced by 86.2%. At lower concentrations, B. bassiana was not effective against S. oryzae. The study suggested the need for higher numbers of conidial inoculation for maximum mortality. Kulagod et al. (2001) evaluated insecticides against Yellow Stem Borer [Scirpophaga incertulas] and Leaf folder [Cnaphalocrocis medinalis] on rice was in Uttara Kannada, Karnataka, India, during the Kharif 2008: fipronil 5 FS (2.5 ml), indoxacarb 14.5 SC (0.5 ml), thiodicarb 75 WP (1.0 g), Bacillus thuringiensis (1.0 g), novaluron 10 EC (0.1 ml/litre), cartap hydrochloride 50 SP (1.0 g/litre), spinosad 45 SC (0.2 ml/litre), chlorpyrifos 20 EC (2.0 ml/litre), flubendiamide 480 SC (0.2 ml/litre) and azadirachtin (5000 ppm). At 5 days after spraying (DAS), the chemical treatments were on a par and superior to B. thuringiensis, azadirachtin and control in reducing dead heart incidence; almost a similar trend was observed at 10 DAS. At 15 DAS, cartap hydrochloride 50 SP, fipronil 5 FS and flubendiamide 480 SC registered the lowest percent dead heart. Cartap hydrochloride 50 SP (4.83%), fipronil 5 FS (4.99%), flubendiamide 480 SC (4.12%) and thiodicarb 75 WP (5.58%) recorded the lowest percent white ear. The treatments were at par in terms of damaged leaves at 5 DAS. At 10 and 15 DAS, flubendiamide 480 SC, spinosad 45 SC, indoxacarb 14.5 SC and fipronil 5 FS resulted in the lowest percentage of damaged leaves. Cartap hydrochloride 50 SP gave the lowest percent dead heart and 34

12 white ear incidence, and the highest grain yield. However, this treatment was on a par with fipronil 5 FS, flubendiamide 480 SC and thiodicarb 75 WP for these traits. Misra et al. (2001) evaluated in the field experiment twenty-seven rice cultivars for growth performance, and pest and disease resistance in Uttar Pradesh, India during Nilaparvata lugens and Nephotettix virescens were below moderate levels on all cultivars except Suryu-52, IRRI-137, MTU-1001 and Nagarjuna. Yellow stem borer (Scirpophaga incertulas) incidence was low to moderate in Pusa-basmati, IRRI-123, Basmati and Nagarjuna, but was trace to low in all other cultivars. Incidences of brown leaf spot (Drechslera oryzae [Cochliobolus miyabeanus]), bacterial leaf blight (Xanthomonas campestris), Cercospora leaf spot (Cercospora oryzae) and false smut (Ustilaginoidea virens) were also observed. The occurrence of biological control agents and natural enemies in these cultivars was also recorded. Padmaja and Kaur (2001) recorded for the first time the pathogenicity of entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorokin has been on rice leaf folder, Cnaphalocrocis medinalis (Guenee) (Lepidoptera: Pyralidae). Effective control of the pest was recorded under field conditions after application of spore suspension of Metarhizium anisopliae in gelatin (1%) at 1x10x spores/ml on the infested rice crop. Between 5 to 7 days after treatment, per cent mortality was recorded. Vijila et al. (2001) evaluated in laboratory bioassays the mortality of rice weevil, Sitophilus oryzae (L.) caused by Metarhizium anisopliae (Metschnikoff) Sorokin alone and in combination with a neonicotinoid insecticide, acetamiprid. Spray application of M. anisopliae alone at a spore concentration of 6x107 conidia/ml required 6 days to cause 10 percent mortality whereas 4x107 conidia/ml required 8 days to cause 10 percent mortality. Acetamiprid alone at a concentration of micro g/dl required 96 h to cause 10% mortality and at micro g/dl required 120 h to cause 20% mortality. In combination with acetamiprid, M. anisopliae killed S. oryzae significantly faster than without acetamiprid. M. anisopliae at a spore concentration of 3x107 conidia/ml with 7.50 micro Niazi et al. (2002) studied the Beauveria bassiana and Paecilomyces amoeneroseus against rice green leafhopper Nephotettix virescens. In the first experiment, adult leafhoppers were separately sprayed in 2 batches with spore suspension. In the second 35

13 experiment, 4 rice seedlings (cv. TN-1) were separately sprayed with spore suspension. The in vivo effect of the entomogenous fungi on the survival of leafhoppers were also studied by allowing the leafhopper to feed for 22 h on spore suspension mixed with 10% sucrose solution enclosed in parafilm M membrane (stretched 4x). In the first experiment, P. amoeneroseus (100% mortality in 20 days) was more effective than B. bassiana. Treatment of rice seedlings with B. bassiana spore suspension, in the second experiment, was most effective (100% mortality in 20 days). Alice et al. (2003) revealed that natural mycosis occurred in the pests of rice during Rabi. In rice, the leaffolder (Cnaphalocrocis medinalis) and the skipper (Pelopidas mathias) were infected and mummified by the fungus. The cadavers were covered by a white powdery mycelium. Mortality was highest in the first and second larval instars. Microscopic examination of the dead larvae showed a large number of spores. Spores were single-celled borne on zigzag conidiophores found in clusters. The pathogenic fungus was identified as Beauveria bassiana. Amaugo and Emosairue (2003) conducted field experiment during the 1999 and 2000 wet season at the National Cereal research Institute, Amakama sub-station, Umuahia, Nigeria to evaluate the efficacy of aqueous and acetone extracts of some indigenous medicinal plants for the control of upland Rice Stem Borers. The treatment consisted of 2.55, 5 and 10% each of aqueous and acetone extract of seed kernels of neem (Azadirachta indica, A. Juss), nutmeg Mondoora myristica (Gaertn.) Dunal, Physic-nut (Jatropha curcas, L.), castor oil (Ricinus communis L.) synthetic insecticides: monocrotophos and the untreated check. The result showed that the monocrotophos and a number of the plants seed kernel extract treatment significantly (P<0.05) reduced the % deadhearts and % whiteheads caused by stem borer. The yield of the rice culture in the two years trails were significantly (P>0.05) higher in monocrotophos treated plants and in a number of the plants extracts treated plant than the control. Comparatively, neem seed and physic nut seed kernel extracts were superior to the other plant extract in controlling stem borers and influencing yield of the crop. The result suggest, that aqueous and acetone extract of neem seed kernel and physic nut seed kernel can be sued by upland rice farmers to control stem borers and obtain higher crop yields. 36

14 Keller and Tuor (2003) isolated 50 different fungal strains from the soil by dilution plate and Galleria mellonella baiting methods. A few strains (Nomuraea sp.) were isolated from naturally infected Spodoptera found on sugarbeet. Using a bioassay with Helicoverpa armigera larvae, 4 promising strains (2 strains of Metarhizium sp., one strain of Nomuraea sp. and one strain of Beauveria sp.) were selected on the basis of mortality (>70% in 7 days) for further large scale production and field trials. The biochemical characterization with regard to cuticle degrading enzymes, effect of different environmental conditions on the spore germination, and growth on solid substrates, such as beaten rice, sorghum, wheat and barley for maximum sporulation, were also studied. Rao et al. (2003) studied efficacy of various insecticides and biopesticides in controlling Cnaphalocrocis medinalis infesting rice cv. Pusa 834. The treatments included: phorate at 0.5 and 0.75 kg a.i./ha; thiamethoxam at 12.5 g a.i./ha; thiamethoxam + Dispel L (Beauveria bassiana) at 12.5 g a.i./ha ml; profenofos at 750 g a.i./ha; profenofos + Biobit DF (Bacillus thuringiensis) at g a.i./ha; lambda-cyhalothrin at 25 g a.i./ha; lambda-cyhalothrin + Biobit DF at g a.i./ha; flufenoxuron at 100 g Biobit + flufenoxuron at g a.i./ha; Dispel at 1000 ml; Biobit DF + Dispel L at 750 g ml; Vitex negundo leaf extract at 1%; neem oil at 1%; Datura sp. whole plant extracts at 1%; Chukrasia velutina [Chukrasia tabularis] seed extract at 1%; solvent + emulsifier; and the control. The lowest leaf folder damage was observed with flufenoxuron and lambda-cyhalothrin treatments, but these insecticides adversely affected the predator populations (Coccinellids and spiders). Biobit treatment resulted in a leaf folder damage of 17.55% folded leaves, and in the highest predator population in the range predator/five hills. Shahid et al. (2003) applied the Bt (Bacillus thuringiensis) at 250 kg/acre and CAMB fungi (Metarhizium anisopliae) at 250 kg/acre, applied alone or in combination, for controlling stem borer (Scirpophaga incertulas) and leaf folder (Cnaphalocrocis medinalis) infesting rice in field trials in Pakistan. Bioassays were also conducted in the laboratory to investigate the pathogenicity of the fungal suspension or bt formulation on Helicoverpa armigera larvae after 48, 72 and 96 h of exposure. Bioassay results showed that the larval mortality was 65, 75 and 100% after 48, 72 and 96 h, respectively. In field trials, CAMB pesticides reduced insect population on rice from 1 to 9 days after spraying. 37

15 Yasodha and Narayanasamy (2004) isolated three entomopathogenic fungi viz., Mucor hiemalis, Fusarium moniliforme [Gibberella fujikuroi] and Scopulariopsis sp. from the yellow rice borer (Scirpophaga incertulas), Leaffolder cutworm (Cnaphalocrocis medinalis) and Yellow hairy caterpillar moths of rice ecosystem. Geng and Zhang (2005) studied the effects of low concentration buprofezin and M. anisopliae var. acridum on N. lugens nymphs (young and old) and adults. The treatments included: buprofezin at 2.5 micro g/ml; fungi at 107 conidia/ml; and combination of buprofezin + fungi. The combination of insecticide + fungi was effective in increasing insect mortality. Nguyen et al. (2005) conducted the experiments in the greenhouse and field to evaluate the efficacy of some new isolates of Metarhizium anisopliae and Beauveria bassiana against Rice Ear Headbug (REB), Leptocorisa. acuta. The results in greenhouse showed that all of 12 selected isolates of M. anisopliae and B. bassiana which have been isolated from naturally infected insects during were found to be pathogenic to the tested Rice Ear Headbug (REB). The mortality percentage of L. acuta caused by B. bassiana and M. anisopliae isolates ranged from 57.5 to 77.7% and from 74.7 to 87% at 10 DAT, respectively. In field experiments, all of 12 selected isolates of M. anisopliae and B. bassiana were found to be effective for controlling REB; the efficacy could be seen from 7 DAT and reached to its highest peak at 14 DAT. The field mortality of L. acuta caused by B. bassiana and by M. anisopliae isolates ranged from 45.3 to 74.9% and from 63.6 to 86.6% at 10 DAT, respectively. M. anisopliae showed better efficacy against REB as compared to B.bassiana. Among 12 new selected isolates of M. anisopliae and B. bassiana have been tested, M.a (OM 3 -BD), M.a (HG 3 -B) and M.a (HG 5 -BD) exhibited higher efficacy against the REB as compared to the rest. Rupesh et al. (2005) evaluated efficacy of B. bassiana against D. armigera infesting rice (cv. Taraori Basmati) in a pot experiment. B. bassiana suspension (106 conidia/ml) was sprayed to seedlings, and 10 adults of D. armigera were released per pot at 0, 12, 24, 36 and 48 h after spraying. Adult mortality was evaluated at 3, 6, 9 and 12 days after releasing the insects. The efficacy of B. bassiana decreased with the increase in the period between fungal inoculation and adult release. On the 12th day, mean percent mortality was highest for adults released at 0 (93.3%) and 12 h (83.3%) after spraying. 38

16 The lethal time mortality of D. armigera caused by B. bassiana was lowest (5.50 days) when adult release was simultaneous with fungal inoculation. Anandhi and Pillai (2006) revealed that the percentage of infection by entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana, varied significantly in relation to the age of the rice crop. Mortality of the black bug, Scotinophara coarctata, due to infection by M. anisopliae and B. bassiana was first observed at 16 days after transplanting (DAT) and peaked at 72 DAT. Gindin et al. (2006) reported that the red palm weevil (RPW, Rhynchophorus ferrugineus) (Olivier) (Coleoptera: Curculionidae) was one of the most severe pests of various palm species, including date palms. While examining the susceptibility of RPW to two entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana, strains of the former were found to be more virulent than those of the latter, achieving 100% larval mortality within 6-7 days. The most virulent strains of M. anisopliae were then tested on RPW eggs and adults. Incubation in a substrate treated with M. anisopliae spores increased egg mortality and reduced their hatchability. The total percentage mortality of eggs and hatched larvae was 80-82%, compared with 34% in the controls. RPW adults were challenged with two types of fungal formulation: dry powder and aqueous suspension. Cumulative adult mortality of 100% was achieved in 2-3 weeks for the dry rice-based formulation and in 4-5 weeks for the spore suspension. Patel et al. (2006) tested the isolates of Beauveria bassiana (Balsamo) Vuillemin tested for biological control of rice stink bug, Oebalus pugnax (F.), in the laboratory, in smallplot field experiments and compared with conventional insecticides, and in a large-plot experiment to determine the spread and persistence of the fungus. Isolate RSB was found in a naturally-infected O. pugnax in a rice field near Crowley, LA, and isolates LRC28 and LRC21 were obtained for their relatively good growth at high temperatures. The soilderived isolate LRC28 was more virulent to O. pugnax adults than isolate RSB in a laboratory experiment. The fungal isolates, applied at conidia/ha, did not differ from one another in reducing insect numbers or in infecting rice stink bugs in the small-plot experiments, although isolates LRC28 and RSB, but not LRC21, occasionally differed from the control. 39

17 Rao et al. (2006) tested the various combinations of insecticides (phorate at 0.5 and 0.75 kg/ha, thiamethoxam 12.5 g/ha, profenofos at 750 g/ha, lambda-cyhalothrin at 25 g/ha and flufenoxuron at 100 g/ha), botanicals (Vitex negundo 1%, neem oil 1%, Datura sp. 1% and Chukrasia velutina 1%) and biopesticides (Biobit [Bacillus thuringiensis subsp. kurstaki] at 750 g/ha. and Dispel (Beauveria bassiana) at 1000 ml/ha.) for their effects on Rice leaffolder (Cnaphalocrocis medinalis) and its coleopteran predators (Paederus fuscipes and Ophionia indica [Casnoidea indica]). Sarao and Mehal (2006) evaluated the effectiveness of different level of nitrogen with the insecticide against insect pest of rice during kharif 2005 and 2006 at village balloke (Ludhiana). The pooled mean of both the year showed that leaf folder infestation (10.88), dead heart (4.73%), white year (4.98%) infestation and plant hopper population were significantly more in dose of higher level of N (375kg urea/ha), grain yield in case of 225, 275 and 325 kg urea per ha was q/ha. Ambethgar et al. (2007) evaluated pathogenicity of 42 local isolates of entomopathogenic fungi including Beauveria bassiana (twenty two), Meturhizium anisopliae (three), M. fluvoviride (one), Nomuraea rileyi (four), Paecilomyces sp. (one), Aspergillus spp. (five), Fusarium spp. (three), Zoophthora radicans (two) and Entomophthora sp. (one) originating from a range of insect species against third instar larvae of Cnaphalocrocis medinalis. All the isolates tested were pathogenic to the pest at varying degrees. In single-dose (1x107 conidia/ml) time-mortality assay, only five isolates of B. bassiana, viz., BbCm KKL 1100, BbCm TVR 0101, BbCm ADT 0101 (isolated from C. medinalis), BbMp KKL 1195 (isolated from Marasmia patnalis) and BbOn KKL 0597 (isolated from Oxya nitidula), were superior to all other isolates with BbCm KKL 1100 having the lowest LT50 value of 7.81 days. In the multiple dose assays (ranging from to conidia/ml), the Karaikal isolate of B. bassiana (BbCm KKL 1100 isolated from C. medinalis) was found to be the most virulent. The LC50 value estimated at 13 days post-inoculation from three independent bioassays for this isolate was 2.8 x103 conidia/ml. Majidi-Shilsar (2007) collected fungal pathogens of the rice striped stem borer, C. suppressalis, from dead larvae in rice fields in Guilan province, Iran. The isolated fungi were incubated with larvae placed on wet sterile filter paper, and incubated at 25 (+) 2 C 40

18 and relative humidity of 85 (+) 5 %. The fungal colony was formed on dead larvae within h, and sporulation started after 5-7 days. The fungi appeared on the larvae as white or green colonies. The fungi were identified as B. bassiana and M. anisopliae. The spores of B. bassiana were hyaline, and globose to subglobose ( micro m), while the spores of M. anisopliae were cylindrical to oval, often slightly narrower in the middle, and truncate at both ends ( micro m). Rabindra and Ramanujam (2007) reported that the sucking pests cause serious damage to several agricultural, horticultural and plantation crops either by direct feeding or by transmitting plant viral diseases. Since sucking pests like plant and leaf hoppers, aphids, whiteflies, scale insects, thrips and mites to have developed resistance to insecticides, biological control using microbial pathogens, particularly fungal pathogens like Beauveria bassiana, Metarhizium anisopliae, and Verticillium lecanii, has been explored for a number of pests. Several commercial formulations based on entomopathogenic fungi were developed for the control of sucking pests in different countries. Mycotrol and Botanigard based on B. bassiana, Mycotal based on V. lecanii and PFR-97 and Pae-Sin based on Paecilomyces fumosoroseus were developed for the control of whiteflies, aphids and thrips. Dhuyo et al. (2008) evaluated two isolates (274 and 373) with concentrations ranging from 105 to 109 spores ml-1 of the entomopathogenic fungus Beauveria bassiana (Bal.) Vuill for pathogenicity against immature stages of yellow rice stem borer, Scirpophaga incertulas (Walker) under laboratory conditions at Rice Research Institute, Dokri, Sindh Pakistan. Not only the highest rates of mortality of S. incertulas were recorded at higher concentrations of spores, ml-1 but also reduced percentage egg hatching of yellow rice stem borer. Log dose (spores/ml) of the fungus B. bassiana isolate No. 274 and isolate No.373 on egg, larva and pupa of S. incertulas was recorded as 5.12, 5.46, 6.10 and 6.23, 8.33, 8.33, respectively. The isolate No.274 was more pathogenic than the isolate No.373 to control of S. incertulas. Jin et al. (2008) studied the effect of fungal isolates for microbial control of brown planthopper (BPH) Nilaparvata lugens (Stal), to which little attention has been paid in the past two decades. Results: Thirty-five isolates of Metarhizium anisopliae (Metschnikoff) Sorokin and M. flavoviride Gams & Rozsypal from different host insects 41

19 worldwide were bioassayed for their lethal effects against third-instar BPH nymphs at 25 C and a 14:10 h light:dark photoperiod at ca 1000 conidia mm-2. On day 9 posttreatment, mortality attributable to mycosis ranged from 6.5 to 64.2% and differed significantly among the tested isolates with no apparent relationship to their host origin. Only two BPH-derived M. anisopliae isolates from the Philippines (ARSEF456) and Indonesia (ARSEF576) killed >50% of the nymphs. Both isolates were further bioassayed for time-concentration-mortality responses of the nymphs to the sprays of 19-29, and conidia mm-2 in repeated bioassays. The resultant data fitted a time-concentration-mortality model very well. Their LC50 values were estimated as 731 and 1124 conidia mm-2 on day 7 and fell to 284 and 306 conidia mm-2, respectively, on day 10. Conclusion: The two M. anisopliae isolates are potential biocontrol agents of BPH for further research. This is the first report of the lethal effects of global Metarhizium isolates on the rice pest. Sher et al. (2008) studied the effects of cartap hydrochloride 4 G at 0.75 kg a.i./ha, Beauveria bassiana at 2.5 kg/ha, Bacillus thuringiensis at 1.5 kg/ha, imidacloprid 17.8 SL at 0.05%, Trichogramma japonicum at eggs/ha, Neem Gold at 5 ml/litre and Neem Gold at 3 ml/litre + T. japonicum at eggs/ha on yellow stem borer (YSB; Scirpophaga incertulas) and rice leaf folder (Cnaphalocrocis medinalis) infesting rice (cv. Basmati), and their predators (spiders). Cartap hydrochloride 4 G and imidacloprid 17.8 SL resulted in the lowest incidence of yellow stem borer and leaf folder, but had adverse effects on spider fauna. Bacillus thuringiensis and Beauveria bassiana were the most effective against both pests. The biological pesticides were superior over the synthetic insecticides in conserving the spider population. The release of T. japonicum with botanicals reduced the populations of both pests and conserved the spider population. The cost: benefit ratio was highest with imidacloprid 17.8 SL (1:10.13) and lowest with Bacillus thuringiensis (1:1.90). Ambethgar et al. (2009) studied the compatibility of Beauveria bassiana (Balsamo) Vuillemin (isolate BbCm KKL 1100) with twelve insecticides and three neem formulations on agar plate to develop suitable combinations for the management of insect pests in rice fields. All chemical and botanical insecticides inhibited mycelial growth of B. bassiana either partially or completely depending on their concentrations (10X, 1X 42

20 and 0.1X, where X=treated concentration). Chemical insecticides completely inhibited the mycelial growth of B. bassiana, while the neem formulations inhibited 70-86% biomass production of the fungus at 10X concentration. At 1X concentration, carbofuran caused total inhibition, but all other insecticides caused % inhibition. However, at 0.1X concentration, only neem seed kernel extract, chlorpyriphos and dimethoate exhibited 22.2%, 27.3% and 32.6% mycelial inhibition, respectively, and these could be used with B. bassiana in the field condition. Kalita et al. (2009) evaluated the bioefficacy of six new biopesticides (three entomopathogenic fungi and three neem formulations) viz., Beauveria bassiana (BiopowerReg.), Metarhizium anisopliae (BiomagicReg.), Verticillium lecanii (Verticel), NimbicidineReg., MultineemReg. and AchokReg. against insect pests of rice during 2007 and 2008 at ICAR Research Complex for NEH Region, Sikkim Centre, Tadong with monocrotophos as check. All the insecticides were found effective in suppressing pests population and consequently increase in yield (33.99 to q/ha). In comparison to check (monocrotophos), biopesticides were inferior in reducing pest population. Among the biopesticides, 3 ml/1 was found most effective followed by Achook at same dose in managing stem borer, whorl maggot and leaf folder. M. anisopliae (BiomagicReg.) and B. bassiana (BiopowerReg.) were found effective against gundhi bug, damaged grain 4.89% and 5.15% respectively in comparison to untreated control (13.06%). Karthikeyan and Jacob (2009) studied the efficacy of the white muscardine fungus, Beauveria bassiana (Balsamo) Vuillemin and the entomopathogenic nematode, Heterorhabditis indica (Poinar) against adult and grubs of rice blue beetle, Leptispa pygmaea Baly. B. bassiana caused a cumulative adult mortality of % at spores ml-1 and the LC50 value was spores ml-1. The entomopathogenic nematode, H. indica caused a cumulative mortality of % at concentrations of 5IJs to 9IJs in the grubs of L. pygmaea. The cumulative LC50 value was 3.83IJs. The mortality of L. pygmaea was found to increase with reducing dose and increasing exposure period of both B. bassiana and H. indica. Dharmadasa et al. (2010) reported the coffee berry borer Hypothenemus hampei (Ferrari) was the most frequently occurring and destructive pest of coffee (Coffea spp.) in 43

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