Isolation, Molecular Identification and Antibacterial Activity of Endophytic Bacteria from the Bark of Plumeria acuminata

Authors

Lalu Zulkifli , Dewa Ayu Citra Rasmi , AA Sukarso , Yayuk Andayani , Dwi Soelistya Dyah Jekti

DOI:

10.29303/jppipa.v8i3.2249

Published:

2022-07-31

Issue:

Vol. 8 No. 3 (2022): July

Keywords:

Endophytic bacteria, P. acuminata, Inhibition zone, 16S rDNA

Research Articles

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Zulkifli, L., Rasmi, D. A. C. ., Sukarso, A. ., Andayani, Y. ., & Jekti, D. S. D. . (2022). Isolation, Molecular Identification and Antibacterial Activity of Endophytic Bacteria from the Bark of Plumeria acuminata. Jurnal Penelitian Pendidikan IPA, 8(3), 1158–1165. https://doi.org/10.29303/jppipa.v8i3.2249
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Abstract

Endophytic bacteria are able to produce bioactive compounds that are important for applications in the medical field. This study aims to isolate, identify and test the antibacterial activity of endophytic bacteria from the bark of Plumeria acuminata growing in the Mataram area. Isolation was carried out using four types of media, namely NA, TSA, MAC, and BHI. The well diffusion technique was used to test the inhibitory ability of the endophytic bacterial extract against the test pathogenic bacteria. The four test bacteria used were S. aureus, B. cereus, P. aeruginosa, K. pneumoniae. Isolates that showed inhibitory activity were then identified morphologically, biochemically and molecularly based on the 16S rRNA gene. The results of the isolation process obtained 35 colonies. where 9 of them inhibit the growth of test bacteria. There were 2 isolates that able to inhibit the growth of 4 types of test bacteria with an average clear zone diameter of very strong category (21–30 mm), namely isolates T1 and T2. The N2 isolate had the highest average inhibition zone compared to all isolates of endophytic bacteria, which was 32 mm, but was specific for P. aeruginosa. Two isolates, namely B1 and M3 gave various growth inhibitory effects with strong to very strong categories. Other isolates showed various antibacterial effects against at least 2 types of test bacteria. S. aureus was the most resistant pathogenic bacteria to the treatment of endophytic bacterial isolate extract. The cell shape of all endophytic isolates was streptobacillus with or without spores, and 50% of them were Gram-positive. Molecular identification based on the 16S rDNA gene grouped the 9 isolates into 2 main clusters, namely the genus Bacillus and the genus Pseudomonas. Isolates that were closely related to the genus Bacillus showed higher inhibitory activity than those of Pseudomonas. The results showed that isolates of endophytic bacteria from the bark of P acuminata have the potential as an important source of antibacterial substances.

References

Ardila, B. (2013). Uji Daya Hambat Getah Bunga Kamboja (Plumeria acuminata) Terhadap Pertumbuhan Shigella dysentri Secara in vivo. Jurnal Analis Kesehatan Klinical Sains. 1(1). 1-8.

Ashraf, M.D.F., Mazumder, A., Shambhawee, S., Mazumder, R., (2012). Review on Plumeria acuminata. International Journal of Reseach in Pharmacy and Chemestry (review Article) IJRPC 2(2). 467-469

Casella, T. M., Eparvier, V., Mandavid, H., Bendelac, A., Odonne, G., Dayan, L., Duplais, C., Espindola, L. S., & Stien, D. (2013). Antimicrobial and cytotoxic secondary metabolites from tropical leaf endophytes: Isolation of antibacterial agent pyrrocidine C from Lewia infectoria SNB-GTC2402. Phytochemistry, 96, 370–377. https://doi.org/10.1016/j.phytochem.2013.10.004

Ebadi, M. (2007). Pharmacodynamic Basis of Herbal Medicine.2nd Ed. CRC Press. Boca Raton

Gandhi, S. G., Mahajan, V., & Bedi, Y. S. (2015). Changing trends in biotechnology of secondary metabolism in medicinal and aromatic plants. Planta, 241(2), 303–317. https://doi.org/10.1007/s00425-014-2232-x

Gupta, M. Mazumder, UK. Gomathi, P. Thamil, S. (2008). Antimicobial activity of methanol extracts of Plumeria acuminata Ait. Leaves. Natural Product Radiance. 7(2). 102-105. Retrieved from http://nopr.niscair.res.in/handle/123456789/5651

Gupta, M., & Yadav, N. (2016). Phytochemical screening of leaves of Plumeria alba and Plumeria acuminata. Journal of Chemical and Pharmaceutical Research, 8(5), 354-358. Retrieved from https://www.jocpr.com/abstract/phytochemical-...6498.html

Hallmann J., Quadt-Hallmann A., Mahaffee WF. And Kloepper JW. (1997). Bacterial endophytes in agricultural crops. Can. J. Microbiol., 43. 895-914. https://doi.org/10.1139/m97-131

Kusari, S., Pandey, S. P., & Spiteller, M. (2013). Untapped mutualistic paradigms linking host plant and endophytic fungal production of similar bioactive secondary metabolites. Phytochemistry, 91, 81-87. https://doi.org/10.1016/j.phytochem.2012.07.021

Levy, S. B. (2002). Factors impacting on the problem of antibiotic resistance. Journal of Antimicrobial Chemotherapy, 49(1), 25-30. https://doi.org/10.1093/jac/49.1.25

Lin, T. P., Chen, C. L., Chang, L. K., Tschen, J. S., & Liu, S. T. (1999). Functional and transcriptional analyses of a fengycin synthetase gene, fenC, from Bacillus subtilis. Journal of bacteriology, 181(16), 5060–5067. https://doi.org/10.1128/JB.181.16.5060-5067.1999

Liu J. Zhang Z., Zhu H., Dang H., Lu J., Cui Z., (2011). Isolation and characterization of amylase from marine Pseudomonas sp. K6-28-040. African J Biotech, 10(14), 2733-2740. http://dx.doi.org/10.5897/AJB10.2042

Long, H. H., Schmidt, D. D., & Baldwin, I. T. (2008). Native bacterial endophytes promote host growth in a species-specific manner; phytohormone manipulations do not result in common growth responses. PloS one, 3(7), e2702. https://doi.org/10.1371/journal.pone.0002702

Ludwig-Müller J. (2015). Plants and endophytes: equal partners in secondary metabolite production?. Biotechnology letters, 37(7), 1325–1334. https://doi.org/10.1007/s10529-015-1814-4

Mishra, A., Singh, S. P., Mahfooz, S., Singh, S. P., Bhattacharya, A., Mishra, N., & Nautiyal, C. S. (2018). Endophyte-Mediated Modulation of Defense-Related Genes and Systemic Resistance in Withania somnifera (L.) Dunal under Alternaria alternata Stress. Applied and environmental microbiology, 84(8), e02845-17. https://doi.org/10.1128/AEM.02845-17

Morales, G., Sierra, P., Mancilla, A., Paredes, A., Loyola, L. A., Gallardo, O., & Borquez, J. (2003). Secondary metabolites from four medicinal plants from northern Chile: antimicrobial activity and biotoxicity against Artemia salina. Journal of the Chilean Chemical Society, 48(2), 13-18. http://dx.doi.org/10.4067/S0717-97072003000200002

Mursito B., & Prihmantoro, H. (2011). Tanaman Hias Berkhasiat Obat, Ed ke-4. Jakarta: Penebar Swadaya.

Nazar, S., Ravikumar, S., Williams, G. P., Ali, M. S., & Suganthi, P. (2009). Screening of Indian coastal plant extracts for larvicidal activity of Culex quinquefasciatus. Indian J Sci Technol, 2(3), 24-27. http://dx.doi.org/10.17485/ijst/2009/v2i3/29407

Ngamau CN., Matiru VN., Muthuri CW., & Tani A., (2012). Isolation and identification of Endophytic Bakteria of bananas (Musa spp.) in Kenya and Their potential as biofertilizer for sustainable banana production. African Journal of Microbiology Research. Vol 6(34) pp: 6414-6422. Retrieved from https://www.worldagroforestry.org/publication/isolation...and-their

Sahu, S., Chaturvedi, R., Mathew, B., Behra, P., Divya, R., Venketesha, R. T., & Sadanandan, B. (2014). Antibacterial and antioxidant activity of endophytic bacteria isolated from Annona muricata. Int. Rev. Appl. Biotechnol. Biochem, 2, 179-188

Singh, S. P., and Gaur, R. (2017). Endophytic Streptomyces spp. Underscore induction of defense regulatory genes and confers resistance against Sclerotium rolfsii in chickpea, Biological Control 104, 44–56. https://doi.org/10.1016/j.biocontrol.2016.10.011

Sriwarthini, N.L.P.N. (2014). Uji aktivitas antibacteri ekstrak kulit batang kamboja (P.acuminata) terhadap bakteri isolat klinik, FKIP Universitas Mataram : Mataram

Strobel, G.A. and Daisy, B. (2003). Bioprospecting for Microbial Endophytes and Their Natural Products. Microbiology and Molecular Biology Reviews, 67, 491-502. https://doi.org/10.1128/MMBR.67.4.491-502.2003

Sunkar, S. dan Nachiyar, C.V. (2013). Isolation and Characterization of an Endophytic Bacterium from Brassica oleracea with Potential Enzyme and Antibacterial Activity, Asian Journal of Pharmaceutical and Clinical Research, 6(2). 183-187.

Tiwari, P., & Bae, H. (2020). Horizontal Gene Transfer and Endophytes: An Implication for the Acquisition of Novel Traits. Plants (Basel, Switzerland), 9(3), 305. https://doi.org/10.3390/plants9030305

Tjitrosupomo G. (2000). Taksonomi Tumbuhan obat-obatan. Gajah Mada University Press: Yogyakarta

Wahyudi, & Sukarjati. (2013). Pengaruh ekstrak etil asetat getah kamboja (Plumeria acuminate) Terhadap pertumbuhan dan daya hambat bakteri Staphylococcus aureus. Universitas PGRI Adi Buana: Surabaya

Yahya, M., & Hindiah, M. H. (2014). Antibacterial activities of ethyl acetate extract from Plumeria alba stem bark. Scientific Study & Research. Chemistry & Chemical Engineering, Biotechnology, Food Industry. 15(2). 107.

Yao, S., Gao, X., Fuchsbauer, N., Hillen, W., Vater, J., & Wang, J. (2003). Cloning, sequencing, and characterization of the genetic region relevant to biosynthesis of the lipopeptides iturin A and surfactin in Bacillus subtilis. Current microbiology, 47(4), 272–277. https://doi.org/10.1007/s00284-002-4008-y

Vasanthakumari, R. (2009). Practical Microbiology. New Delhi, BI Publications Pvt Ltd.

Author Biographies

Lalu Zulkifli, SCOPUS ID : 15066284800, Mataram University, Faculty of Learning Teacher and Education Science, Indonesia

Dewa Ayu Citra Rasmi, University of Mataram

AA Sukarso, Universitas Mataram

Yayuk Andayani, University of Mataram

Dwi Soelistya Dyah Jekti, University of Mataram

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