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Bio Efficacy of Heterorhabditis Bacteriophora and Oscheius Chongmingensis Against Helopeltis Theivora and Andraca Bipunctata

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Bio Efficacy of Heterorhabditis Bacteriophora and Oscheius Chongmingensis Against Helopeltis Theivora and Andraca Bipunctata Int.J.Curr.Microbiol.App.Sci (2020) 9(4): 2460-2473 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 9 Number 4 (2020) Journal homepage: http://www.ijcmas.com Original Research Article https://doi.org/10.20546/ijcmas.2020.904.295 Bio efficacy of Heterorhabditis bacteriophora and Oscheius chongmingensis against Helopeltis theivora and Andraca bipunctata Bharath Amuri*and Gitanjali Devi Department of Nematology, Assam Agricultural University, Jorhat, Assam, India *Corresponding author ABSTRACT Native isolates of EPNs, Heterorhabditis bacteriophora and Oscheius chongmingensis were isolated from the tea plantation crop of Jorhat district of Assam and were tested against tea mosquito bug Helopeltis theivora andbunch caterpillar, Andraca bipunctata K e yw or ds through mortality test under laboratory condition. For that 5, 10, 50, 100,150, 200 and 250 infective juveniles of each isolate were inoculated per insect. The insect mortality was H.bacteriophora, observed after 24, 48, 72 and 96h exposure. It was observed that 50, 60 and 70 per cent O.chongmingensis, H. theivora and A. mortality of H. theivora were recorded by H. bacteriophora (150 IJs/insect) at 48 h, 72h and 96h of exposure. It recorded that with the increase in the exposure period to 96 h, H. bipunctata bacteriophora could induce up to 90% mortality of the tested insect at the dose of 250 IJs. Article Info Similarly, mortality of H.theivora increased with increase in the dosage of O.chongmingensis and recorded 50, 60 and 70 per cent mortality at 200 IJs/insect at 48 h, Accepted: rd 72 h and 96h. Bioassay of H. bacteriophora and O. chongmingensis against the 3 instar 18 March 2020 larvae of A. Bipunctata indicated that at 250IJs/larva recorded 70 and 60% mortality in 48 Available Online: 10 April 2020 hours of exposure, while 100 and 90% mortality of A. bipunctatawas recorded at 96h.However, in the present investigation, probit analysis revealed that H. bacteriophora showed maximum mortality than O. chongmingensis against H. theivora and A. bipunctata at 48 h,72 h and 96h of exposure time. Introduction health benefits like reduction of weight loss, the risk of diabetes and cardiovascular Tea (Camellia sinensis L.) belonging to the disease. family Theaceae and tribe Gordonaceae is a long duration perennial crop grown under The crop is extensively cultivated in 13 states monoculture. Tea leaves are mostly plucked of India out of which Assam, West Bengal, for the making of tea beverages like green tea Tamil Nadu and Kerala are the largest and black tea and it contain the polyphenols producers. In Assam, it is cultivated on 765 such as catechins and epicatechins. Such tea gardens covering an area of 307.08 compounds act as antioxidant and provide thousands hectares and more than 17% 2460 Int.J.Curr.Microbiol.App.Sci (2020) 9(4): 2460-2473 workers of Assam are engaged in the tea entomopathogenic nematodes (EPNs). industry (Anonymous, 2018).The total annual Entomopathogenic nematodes, Steinernema production of tea in Assam is 676.31 million and Heterorhaditis in the family kg in year 2017-18, which is more than 50 Steinernematidae and Heterorhabditidae of percent of India’s total tea production the order Rhabditida are obligate parasites of (Anonymous, 2018). Crop loss in tea due to insect pests. The other genus Oscheius, in the pests, diseases and weeds varies between 7- family Rhabditidae, is also recently been 15% (Borthakur et al., 1992). Pest infestation isolated and found to be entomopathogenic. is a major problem associated with tea cultivation that caused reduction not only in EPNs are considered as one of the most the quantity but also in quality of tea. Tea significant non-chemical alternatives to insect plantation provides a permanent ecosystem pest control due to their high reproductive for more than1034 arthropods (Chen and potential, ease of mass production and their Chen, 1989). Das (1965) reported that 167 harmlessness to microbes, animals, humans species of arthropods have been noted from and plants. Only the third juvenile stage is the the northeast India and cause 11.00-55.00% infective juvenile that is free-living in the soil, annual yield loss. Among them, tea mosquito non-feeding, encased in a double cuticle with bug, Helopeltis theivora (Waterhouse) closed mouth and anus and capable of (Hemiptera: Miridae) is one of them. surviving for several weeks in the soil, before H.theivora becomes the greatest enemies of infecting a new host individual. tea planters in Africa and Asia causing 55% and 11-100% crop loss, respectively (Wilson The infective juveniles actively penetrate and Clifford 1992; Anonymous, 1994; through the mid gut wall or tracheae into the Sundararaju and Sundara Babu, 1999). In insect body cavity (hemocoel) containing Assam H. theivora caused 15-20% crop loss insect haemolymph. EPNs have a mutualistic (Hazarika et al., 2009; Anonymous, 2010). partnership with gram-negative gamma- This particular pest causes damage to tea proteobacteria in the family plant throughout the year but the incidence is Enterobacteriaceae. Xenorhabdus bacteria are more severe during the months of May- associated with steinernematid nematodes September. while Photorhabdus are symbionts of heterorhabditids. Bunch caterpillars, Andraca bipunctata Walk. (Lepidoptera: Bombycidae) is a well-known Photorhabdus and Xenorhabdus, are carrying pest of tea (Watt and Mann, 1903) and mostly in their intestines, whereas occurs in India, Indonesia, Taiwan, China, Heterorhabditidoides (=Oscheius) carry and Vietnam. In India, the pest is reported symbiotic bacterial strains of Serratia in both from Assam, Sikkim, and West Bengal. The the intestine and the cuticle (Kaya and hatched larvae feed on the shoot, bud, young Gaugler, 1993; Torres-Barragan et al., 2011; leaves, and matured leaves in a group and Zhang et al., 2008, 2009, 2012). completely defoliate the tea plant causing 15% yield loss in Assam (Anonymous, 2010). Nematode and bacteria overcome the insect immune system and the host insect is killed Nematodes which are capable of killing, within 24-48 hours post infection (Adams and sterilizing or hampering the development of Nguyen, 2002). The EPNs thrives in wide insect and completing at least one stage of range of climatic conditions and distributed in their life cycle in the host are called natural and agricultural soils. They have 2461 Int.J.Curr.Microbiol.App.Sci (2020) 9(4): 2460-2473 enormous potential as biocontrol agent juveniles). The Petri dishes were sealed with against a wide range of insect pests but their cling film and stored in polythene bags to pathogencity varied from species to species conserve moisture. The infective juveniles of and /or strain to strain. The application of nematodes were harvested by using White native isolates found to be a prime importance Traps as described by White (1927). The for the management of insect pest of various nematode multiplications in G. mellonella crops because they share common niche with were continued in batches and the infective their host. The present study was undertaken juveniles (IJs) were collected up to 12 days of to evaluate the best native isolates of EPN inoculation. These juveniles were washed and against tea pestlike H. theivora and rinsed several times with sterile distilled A.bipunctata under in-vitro condition through water. The collected IJs were preserved in a bio efficacy test. beaker at 10°C and utilized within two days for the study. Materials and Methods Infectivity assay Collection and preparation of insects The infectivity tests were carried out in cavity Helopeltis theivora and Andraca bipunctata block (55mm x 55mm) lined with a wet filter were collected from the sites in tea (Camellia paper. One larva/adult of test insect was sinensis) plantation areas (26.714910˝N placed in each cavity block. Infective 94.204440˝E) of AAU, Jorhat, Assam, India. juveniles of H. bacteriophora and O. The collected insects used in the experiment chongmingensis were inoculated at 0 were kept in large plastic containers in (control), 10, 50, 100,150, 200 and 250 laboratory at room temperature (25-30°C). /insect. In control the insects were not The adults of H. theivora, having the average exposed to the nematodes. The cavity blocks weight of 6.48±0.13mg and 3rdstage larvae of were wrapped with tiny perforated A. bipunctata average weight of polyethylene bag and kept at room 519.70±2.70mg were separated and used for temperature in a dark room (30±2°C) and infectivity test. replicated 10 times along with control. Maintenance of nematode culture Insect mortality was observed after 24, 48, 72 and 96h exposure. Dead insects were The entomopathogenic nematodes viz., dissected and the hemolymph of each one was Heterorhabditis bacteriophora and Oscheius observed under a stereoscopic microscope. chongmingensis were isolated from tea When nematodes in different stages of through Galleria bating technique and were development were detected, we considered continuously multiply on larva of greater wax they were the cause of death. moth, G. mellonella. The insect larvae were exposed to the nematode as per the method The mortality data (expressed in percentage) described by Woodring and Kaya (1988). were transformed to arc sine and subjected to analysis of variance. The means were About ten larvae of G. mellonella were separated by Duncan’s Multiple Range Test released into a Petri dish over two Whatman (SPSS, 2007). The corrected per cent No.1 filter papers inoculated with infective mortality was computed after considering kill juveniles stored in sterile distilled water (1 ml in control treatment. The LD50 was calculated suspension containing 200 infective using probit analysis and all comparisons 2462 Int.J.Curr.Microbiol.App.Sci (2020) 9(4): 2460-2473 were made at P = 0.05 level of significance The LD50 values determined from different (Hewlett and Placket, 1979). dosage levels for both EPN species against H.
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