Bio Efficacy of Bacillus thuringiensis Isolates against Diamond Back Moth (Plutella xylostella L.) on Cauliflower Plant in Tamil Nadu, India

Main Article Content

S. S. Thilagavathi
G. Prasad
A. Ramalakshmi

Abstract

Bacillus thuringiensis (Bt) gram positive entomopathogenic bacteria being an eco-friendly biopesticide. In present study the potential of B. thuringiensis isolates was studied for the biological control of Diamond back moth (DBM). Fifteen Bt isolates were obtained from the Department of Agricultural Microbiology, TNAU Tamil Nadu. This mainly isolated from the cultivated lands Cotton (Gossypium hirsutum), Brinjal (Solanum melongena) and Tomato (Solanum lycopersicum). All the 15 isolates were identified as B. thuringiensis based on the crystalline structure. Four different types of crystalline forms were observed, in which the isolates CC, CB1, BC, TD, BD were produces cuboidal shape crystals. Then, the isolates were characterized based on presence of lepidopteron specific cry gene. Among the 15 B. thuringiensis isolates seven of them were found to be positive for lepidopteron specific cry genes include cry 1 & cry 2. Four Bt isolates were exhibited presences of both cry 1 and cry 2 genes. The selected 4 isolates further screened for protein profiling by SDS Page. Molecular weights of the protein ranging from 65 to 130 kDa. Toxicity of this four B. thuringiensis isolates were evaluated by bioassay using third instar larvae of the diamondback moth (Plutella xylostella) and isolate (CC) recorded maximum mortality of 95.33% comparable to standard strain HD1 98.31%. In quantitative bioassay, the LC50 for third instar larvae of Plutella xylostella was found to be least range 197.09 ppm with fiducial limits of 110.28 - 352.21 ppm respectively. Different concentration of the toxic protein (100, 250, 500, 750, 1000, 2000) will reduce the leaf damage and larval growth was (54-8% and 58-25%) observed. This achieved effective control of DBM in cauliflower plant.

Keywords:
Bacillus thuringiensis, biopesticide, cry protein, bioassay, Plutella xylostella

Article Details

How to Cite
Thilagavathi, S. S., Prasad, G., & Ramalakshmi, A. (2020). Bio Efficacy of Bacillus thuringiensis Isolates against Diamond Back Moth (Plutella xylostella L.) on Cauliflower Plant in Tamil Nadu, India. International Journal of Plant & Soil Science, 32(1), 10-20. https://doi.org/10.9734/ijpss/2020/v32i130231
Section
Original Research Article

References

Uthamasamy S, Kannan M, Senguttuvan K, Jayaprakash SA. Status, damage potential and management of diamondback moth, Plutella xylostella (L.) in Tamil Nadu, India. The 6th International Workshop on Management of the Diamondback Moth and Other Crucifer Insect Pests. 2005;270-279.

Rashmi G, Sanjay S. Bioecology and management strategy of diamond back moth (Plutella xylostella L.). International Journal of plant protection. 2013;6(1):192-197.

Krishnamoorthy A. Biological control of diamondback moth Plutella xylostella (L.), an Indian scenario with reference to past and future strategies. In Proceedings of the International Symposium. Montpellier France CIRAD. 2004;204-11.

Grzywacz D, Rossbach A, Rauf A, Russel DA, Srinivasan R. Shelton AM. Current control methods for diamondback moth and other brassica insect pests and the prospects for improved management with lepidopteran resistant B. thuringiensis vegetable brassicas in Asia and Africa. Crop Protection. 2010;29:68-79.

Kibata GN. Diamondback moth Plutella xylostella L. (Lepidoptera: Yponomeutidae), a problem pest of brassicae crops in Kenya. In proceeding of the First Biennial Crop Protection Conference. 1996;27-28:1-11.

Wasim AMD, Dwaipayan S, Ashim C. Impact of pesticides use in agriculture: their benefits and hazards. Interdisc Toxicol. 2009;2(1):1–12.

Zhong CH, Ellar DJ, Bishop A, Johnson C, Lin SS, Hart ER. Characterization of a Bacillus thuringiensis δ-endotoxin which is toxic to insects in three orders. Journal of Invertebrate Pathology. 2000;76(2):131–139.

Bravo A, Gill SS, Soberon M. Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon. 2007;49(4):423–435.

Talekar NS. Diamondback moth and other crucifer pests: Proceedings of the sec¬ond international workshop. Asian Vegetable Research and Development Center. 1992; 92:368-603.

Broderick NA, Raffa KF, Handelsman J. Midgut bacteria required for B. thuringiensis insecticidal activity. Proc Natl Acad Sci. 2006;103(41):196–199.

Monnerat RG, Soares CM, Capdeville G, Jones G, Martins ES, Praca L, Cordeiro BA, Braz SV, Dos Santos RC, Berry C. Translocation and insecticidal activity of Bacillus thuringiensis living inside of plants. Microbial Biotechnology. 2009;2(4):512–520.

Kati H, Sezen K, Demirbag Z. Characterization of a highly pathogenic Bt strain isolated from common cockchafer, Melolontha folia. Microbiol. 2007;52(2): 146-152.

Lopez PSA, Martinez JW, Castillo AX, Salamanca JAC. Presence and significance of Bacillus thuringiensis cry protein associated with Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae). Revista de Biologia Tropical. 2009;57(4):1235-43.

Ramalakshmi A, Udayasuriyan V. Diversity of Bacillus thuringiensis isolated from Western Ghats of Tamil Nadu State, India. Current Microbiology. 2010;61:13–18.

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;277: 680-685.

Ahmad T, Ali H, Shfiq M. Biology of diamondback moth, Plutella xylostella (Linn.) on Brassica juncea cv. PUSA BOLD. Asian Journal of Biological Sciences. 2008;3(2):260-262.

Tabashnik BE, Cushing NL, Finson N, Johnson MW. Field development of resistance to Bacillus thuringiensis in diamondback moth (Lepidoptera: Plutellidae). Journal of Economic Entomology. 1990;83:1671–1676.

Bravo A, Sarabia S, Lopez L, Ontiveros H, Abarca C, Ortiz A, Ortiz M, Lina L, Villalobos FJ, Pena G, Valdez MEN, Soberon M, Quintero R. Characterization of cry genes in a Mexican Bacillus thuringiensis strain collection. Applied and Environmental Microbiology 1998;64: 4965–4972.

Lone SA, Yadav R, Malik A, Padaria JC. Molecular and insecticidal characterization of Vip3A protein producing Bacillus thuringiensis strains toxic against Helicoverpa armigera (Lepidoptera: Noctuidae). Canadian Journal of Microbiology. 2016;62(2):179–190.

Senthil NS, Choi MY, Paik CH, Kalaivani K. The toxicity and physiological effect of goniothalamin, a styryl - pyrone, on the generalist herbivore, Spodoptera exigua Hubner. Chemosphere. 2008;72:139-1400.

Finney DJ. Probit Analysis. 3rd edition. Cambridge University Press. Cambridge. 1971;20-63.

Monnerat RG, Masson L, Brousseau R, Pusztai-Carey M, Bordat D, Frutos R. Differential activity and activation of Bacillus thuringiensis insecticidal proteins in Diamondback moth, Plutella xylostella. Current Microbiology. 1999;39:159–162.

Neves AD, Oliveira RF, Parra JRP. A new concept for insect damage evaluation based on plant physiological variables. Annals of the Brazilian Academy of Sciences. 2006;78(4):821–835.

Geeta G, Alagawadi AR, Kridhnaraj PU, Basavana GK. Characterization of Bacillus thuringiensis isolates of Western Ghats and their insecticidal activity against diamond back moth (Plutella xylostella L.). Karnata Journal of Agriculture Sciences. 2012;25(2):199-202.

Babita M, Veena K. Isolation, Characterization and crystal morphology study of Bacillus thuringiensis isolates from soils of Punjab. Journal of Pure and Applied Microbiology. 2018;12(1):189-193.

Rashini YB, Jayanetti K, Ramani R Samaras S, Ovitigala V, Don Si, Jagathpriya, Weerasena, Kurt L, Juan JLF. Identification of a native Bacillus thuringiensis strain from Sri Lanka active against Dipel-resistant Plutella xylostella. Peer J. 2019;7535.

Showkat AL, Abdul M, Jasdeep C, Padaria. Selection and characteriza- tion of Bacillus thuringiensis strains from northwestern Himalayas toxic against Helicoverpa armigera. Microbiology Open. 2017;6:484.

Mahadeva Swamy HM, Selvakumar G, Asokan R, Nagalakshmi G, Soumya BR, Abraham Verghese. Molecular characterization and elucidation of Bacillus thuringiensis Cry1I toxin isolated from the insect pest mango leaf webber, Orthaga exvinacea (Noctuidae: Lepidoptera) Journal of Entomology and Zoology Studies. 2020;8(1):522-530.

Boukedi H, Sellami S, Ktari S, Hassan NBB, Boudawara TS, Tounsi S, Mesrati LA. Isolation and characterization of a new Bacillus thuringiensis strain with a promising toxicity against Lepidopteran pests. Microbiological Research. 2016; (186–187):9-15.

Reyaz AL, Gunapriya L, Indra Arulselvi P. Molecular characterization of indigenous Bacillus thuringiensis strains isolated from Kashmir valley. 3 Biotech. 2017;7:143.

Katiane DSL, Joelma SDS, Maria CDS, Wanderli PT, Ricardo AP Valeria CSP. Isolation and molecular characterization of Bacillus thuringiensis found in soils of the Cerrado region of Brazil and their toxicity to Aedes aegypti larvae. 2018;62.

Praca LB, Caixeta CF, Gomes ACMM, Gomes R, Monnerat. Selection of Brazilian Bacillus thuringiensis strains for controlling diamondback Moth on cabbage in a systemic way. Bt Research. 2013;4(1):1-7.

Kahrizeh AG, Khoramnezhad A, Hassanloui RT. Isolation, characterization and toxicity of native Bacillus thuringiensis isolates from different hosts and habitats in Iran. Journal of Plant Protection Research. 2017;57(3):212–218.

Yılmaz S, Ayvaz A, Akbulut M, Azizoglu U, Karaborklu S. A novel Bacillus thuringiensis strain and its pathogenicity against three important pest insects. Journal of Stored Products Research. 2012;51:33–40.

Ebrahimi MS, Ahad, Reza TH. Effect of Bacillus thuringiensis var. kurstaki on survival and mortality of immature and mature stages of Diadegma insulare parasitizing Plutella xylostella. Phytoparasitica. 2012;40:393–401.

Savitri G, Murali MP. Pathogenicity of the bacterium Bacillus thuringiensis coagulans in silkworm, Bombyx mori (L). Indian Journal of Sericulture. 2003;42(1):4-8.

Torres Q, Mary C, Arenas S, Ivan HV, Victor M, Suarez RR, Pena C. Characterization of Bacillus thuringiensis (Bacillaceae) Strains Pathogenic to Myzus persicae (Hemiptera: Aphididae) Florida Entomologist. 2020;99(4):639-643.

Ferrari JZ, Ana Paula SR, Fernanda A, Pires Fazion, Ana Maria M, Pedro MO, Janeiro N, Laurival A, Vilas B, Gislayne, Trindade VB. Isolation, morphological and molecular characterization of Bacillus thuringiensis strains against Hypothenemus hampei. Revista Brasileira de Entomologia. 2018;62(3):198-204.

Legwailal MM, David C, Munthali, Baone C, Kwerepe, Obopile M. Efficacy of Bacillus thuringiensis (var. kurstaki) against Diamond-back Moth (Plutella xylostella L.) eggs and larvae on cabbage under semi-controlled Greenhouse conditions. International Journal of Insect Science. 2015;7:39–45.

Chandrasekaran R, Revathi K, Jayanthi S. Combined effect of Bacillus thuringiensis and Bacillus subtilis against Helicoverpa armigera. International Journal of Current Microbiology and Applied Sciences. 2017;4 (7):127-141.