Characteristics of Bacillus thuringiensis isolates indigenous soil of South Sumatra (Indonesia) and their pathogenicity against oil palm pests Oryctes rhinoceros (Coleoptera: Scarabaeidae)
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Abstract. Pujiastuti Y, Arsi, Sandi S. 2020. Characteristics of Bacillus thuringiensis isolates indigenous soil of South Sumatra (Indonesia) and their pathogenicity against oil palm pests Oryctes rhinoceros (Coleoptera: Scarabaeidae). Biodiversitas 21: 1287-1294. Bacillus thuringiensis is a gram-positive, entomopathogenic bacterium that could be isolated from soil and be used to control various plant pests. Oryctes rhinoceros is an important pest in oil palm. Application of B. thuringiensis-based bioinsecticides is an alternative in controlling these pests. The purposes of this study were to isolate and identify B. thuringiensis bacteria from the soil of South Sumatra, production of B. thuringiensis-based bioinsecticides and to test their toxicity to O. rhinoceros larvae. The study was conducted in several cities/districts in the province of South Sumatra. Soil samples were taken from various habitats and B. thuringiensis isolates were grown on NGKG agar media. Among 76 soil samples (6 districts and 2 cities) B. thuringiensis colonies were obtained leading to 24 isolates of B. thuringiensis. Toxicity screening tests for armyworm Spodoptera litura were 55.79% (53 isolates) and their mortality to 25.26% O. rhinoceros larvae (24 isolates). From these isolates whose effectively killed O. rhinoceros larvae, 10 isolates were taken and propagated with Nutrient Broth (NB) and biourine enriched with 5% molasses. Number of spores produced was counted during 24, 48 and 72 hours. Furthermore, a bioassay test was carried out on O. rhinoceros larvae for 7 days. Isolate of KJ3P1 caused the highest mortality of O. rhinoceros larvae after 7 days of observation. SDS Page resulted in KJ3P1 and KJ3R5 isolates showing several bands whose content of various types of protein molecular weight. Isolation of B. thuringiensis in South Sumatra produced 2 isolates potentially to be active ingredients in production of bioinsecticides which were effective in killing O. rhinoceros larvae.
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References
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Sarrafzadeh MH. 2012. Nutritional Requirements of Bacillus thuringiensis during different phases of growth, sporulation and germination
evaluated by plackett-burman method. Iran J. Chem. Chem. Eng. 31(4): 131–136.
Soberón M., Rose Monnerat, and Alejandra Bravo. 2018. Mode of Action of Cry Toxins from Bacillus thuringiensis and Resistance
Mechanisms. Chapter. January 2018 DOI: 10.1007/978-94-007-6449-1_28
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Susanto, A, Sudharto, dan AE Prasetyo. 2011. Information of insect pest Oryctes rhinoceros Linn. Research Center of oilpalm. Medan. (in
Indonesian)
Valicente FH, Tuelher EDS, Leite MIS, Freire FL, & Vieira CM. 2010. Production of Bacillus thuringiensis biopesticide using commercial
lab medium and agricultural by-products as nutrient sources. Revista Brasileira de Milho e Sorgo 9(1): 1–11
van Frankenhuyzen, K. 2009. Insecticidal activity of Bacillus thuringiensis crystal proteins. Journal of Invertebrate Pathology 101(1), 1–16.
Wiest S.L.F., Pilz Júnior H.L. , Fiuza L.M. 2015. Thuringiensin: a toxin from Bacillus thuringiensis. Bt Research 2015, Vol.6, No.4, 1-12
trapping and aerial spraying of Bacillus thuringiensis (Bt) for controlling bagworm, Metisa plana Walker (Lepidoptera: Psychidae) in Yong Peng, Johor, Malaysia. Journal of Oil Palm Research 29 (1): 55-65.
Anonim. 2018. Identification of Bacillus species. UK Standards for Microbiology Investigations. Standards Unit Microbiology Services
Public Health England. https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/697260/ID_9i3.1.pdf
Bravo,A., Diana L. Martínez de Castro, Jorge Sanchez. 2015. Mechanism of action of Bacillus thuringiensis insecticidal toxins and their use in the control of insect pests. In book: The Comprehensive Sourcebook of Bacterial Protein Toxins. December 2015 DOI: 10.1016/B978-0-12-800188-2.00030-6
Bravo,A., Isabel Gómez, Gretel Mendoza, Meztli Gaitan, and Mario Soberón. 2015. Chapter 6. Different models of the mode of action of
3d-Cry toxins from Bacillus thuringiensis. December 2015 DOI: 10.1016/B978-0-12-800188-2.00030-6
Bravo,A and M Sobero´ n S S Gill. 2015. Bacillus thuringiensis: Mechanisms and Use. Chapter In Insect Control Biological and Synthetic
Agent. Edited by Lawrence I. Gilbert Sarjeet S. Gill. Academic Press. 42 p.
Brian A. F., Hyun-Woo Park and Dennis K. Bideshi. 2010. Overview of the Basic Biology of Bacillus thuringiensis with Emphasis on
Genetic Engineering of Bacterial Larvicides for Mosquito Control. Open Toxinology Journal, 2010, 3, 154-171
Fernández, J. H. and SA López-Pazos . 2011. Bacillus thuringiensis: Soil microbial insecticide, diversity and their relationship with the
entomopathogenic activity In: Soil Microbes and Environmental Health. Chapter 3. ISBN 978-1-61209-647-6. Editor: Mohammad Miransari, pp. © 2011 Nova Science Publishers, Inc. 24 p.
Federici, Brian A, Park, Hyun-Woo, Bidesh DK. 2010. Overview of the basic biology of Bacillus thuringiensis with emphasis on genetic
engineering of bacterial larvacides for mosquito control. The Open Toxicol J. 3:83-100
He, F. 2011. Laemmli-SDS-PAGE. Bio-101: e80. DOI: 10.21769/BioProtoc.80. June 05, 2011
Jouzani, G.S. & Elena Valijanian & Reza Sharafi. 2017. Bacillus thuringiensis: a successful insecticide with new
environmental features and tidings. Appl Microbiol Biotechnol (2017) 101:2691–2711 DOI 10.1007/s00253-017-8175-y
Khaeruni, R. and Purnamaningrum, 2012. The isolation of Bacillus thuringiensis Berl. from land and its pathogenity to Crocidolomia
binotalis Zell larvae on the Sawi Plant (Brassica juncea L.). J. Agroteknos Maret, 2: 21-27.
Letowski J, Bravo A, Brousseau R, Masson L. 2005. Assessment of cry1 gene contents of B. thuringiensis strains by use of DNA
microassays. Appl Environ Microbiol 71(9):5391–5398
Marzban R. 2012. Investigation on the suitable isolate and medium for production of Bacillus thuringiensis. J. Biopest. 5(2): 144–147.
Mizuki,E., T. Ichimatsu, S.-H. Hwang, Y.S. Park, H. Saitoh, K. Higuchi and M. Ohba. 1999. Ubiquity of Bacillus thuringiensis on
phylloplanes of arboreous and herbaceous plants in Japan. J. of Applied Microbiology, (86): 979–984
Noorhazwani, K; Siti ramlah, A.A., Mohamed Mazmira, M.M; mohd Najib, A; Che Ahmad hafiz,C.M; and Norman K. 2017. Controlling
Metisa plana Walker (Lepidoptera:psychidae) outbreak using Bacillus thuringiensis at an oilpalm plantation in Slim River Perak. Malaysia. J. Oil Palm Res. Vol. 29 (1): 47-54
Osman G. E. H., Rasha Already, A. S. A. Assaeedi, S. R. Organji, Doaa El-Ghareeb, H. H. Abulreesh and A. S. Althubiani. Bioinsecticide
B.thuringiensis a Comprehensive Review. Egyptian Journal of Biological Pest Control, 25(1), 2015, 271-288
Pedro AN, Ibarra JE. 2010. Detection of new cry genes of Bacillus thuringiensis by use of a novel PCR primer system. Appl Environ
Microbiol 76:6150–6155. doi:10.1128/AEM.00797-10
Prabakaran G, Hoti SL, Manonmani AM, & Balaraman K. 2008. Coconut water as a cheap source for the production of endotoxin of
Bacillus thuringiensis var. israelensis, a mosquito control agent. Acta Trop. 105(1): 35–38.
Pujiastuti,Y., D T Astuti, S R Afriyani, S Suparman, C Irsan, E R Sembiring, S Nugraha, Mulawarman and N Damiri. 2018.
Characterization of Bacillus thuringiensis Berl. indigenous from soil and its potency as biological agents of Spodoptera litura (Lepidoptera: Noctuidae). Proceeding IOP Conference Series: Earth and Environmental Science, Vol.102, conference 1
Purnawati R, Sunarti TC, Syamsu K, & Rahayuningsih M. 2014. Characterization of novel Bacillus thuringiensis isolated from Attacus atlas
and its growth kinetics in the cultivation media of tofu whey for bioinsecticide production.J. Biol. Agri. Healthcare 4(16): 33–40.
Rusmana, I and Hadioetomo, R.S. 1994. Isolation of Bacillus thuringiensis Berl. from silkworm farms and their toxicity to the larvae of
Crocodolomia binotalis Zell and spodoptera litura F. Hayati 1 (1): 22-23.
Salazar-Magallon, J.A., Víctor M. Hernandez-Velazquez, Andrés Alvear-Garcia, Iván Arenas-Sosa, Guadalupe Peña-Chora. 2015.
Evaluation of industrial by-products for the production of Bacillus thuringiensis strain GP139 and the pathogenicity when applied to Bemisia tabaci nymphs. Bulletin of Insectology 68 (1): 103-109, 2015 ISSN 1721-8861
Santi, I.S. dan B. Sumaryo. 2008. Effect of the color of pheromone towards number of trapped Oryctes rhinoceros in oilpalm estate. Jurnal
Perlindungan Tanaman Indonesia. 14(2): 76-79. (in Indonesian)
Sarrafzadeh MH. 2012. Nutritional Requirements of Bacillus thuringiensis during different phases of growth, sporulation and germination
evaluated by plackett-burman method. Iran J. Chem. Chem. Eng. 31(4): 131–136.
Soberón M., Rose Monnerat, and Alejandra Bravo. 2018. Mode of Action of Cry Toxins from Bacillus thuringiensis and Resistance
Mechanisms. Chapter. January 2018 DOI: 10.1007/978-94-007-6449-1_28
Sudharto, P. S, dan P. Guritno. 2003. Biological control of oil palm nettle caterpillars in Indonesia : review of research activities in
Indonesia. Oil Palm Research Institute (IOPRI). Proceedings of the PIPOC 2003 International Palm Oil Congress. pp. 362-371.
Susanto, A, Sudharto, dan AE Prasetyo. 2011. Information of insect pest Oryctes rhinoceros Linn. Research Center of oilpalm. Medan. (in
Indonesian)
Valicente FH, Tuelher EDS, Leite MIS, Freire FL, & Vieira CM. 2010. Production of Bacillus thuringiensis biopesticide using commercial
lab medium and agricultural by-products as nutrient sources. Revista Brasileira de Milho e Sorgo 9(1): 1–11
van Frankenhuyzen, K. 2009. Insecticidal activity of Bacillus thuringiensis crystal proteins. Journal of Invertebrate Pathology 101(1), 1–16.
Wiest S.L.F., Pilz Júnior H.L. , Fiuza L.M. 2015. Thuringiensin: a toxin from Bacillus thuringiensis. Bt Research 2015, Vol.6, No.4, 1-12
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