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Sains Malaysiana 53(12)(2024): 3871-3881
http://doi.org/10.17576/jsm-2024-5312-03
Isolation and
Characterization of Endophytic Bacteria from the Vegetative Phase of Indonesian
Garlic var. ‘Tawangmangu Baru’
(Pengasingan dan Pencirian Bakteria Endofit daripada Fasa Vegetatif Bawang Putih Indonesia var. ‘Tawangmangu Baru’)
SAPTINI
MUKTI RAHAJENG1, FARIDA YULIANTI2,*,
UNUN TRIASIH2, DITA AGISIMANTO2, NIRMALA F. DEVY2,
HARDIYANTO2, NUR HAYATI HADIBA3 & HADI M. YUSUF2
1Ketindan Agricultural Training Center, Jln. Ketindan No.1, Lawang, Malang, East Java, Indonesia
Received: 20
March 2024/Accepted: 25 October 2024
Abstract
The high
demand for garlic (Allium sativum L.) has led to efforts to improve its
productivity. Garlic endophytic bacteria have been found to boost the growth of
garlic plants and protect them from diseases without any harmful effects on the
host. The objective of the study was to isolate and characterise the endophytic
bacteria present in the healthy local Indonesian garlic variety ‘Tawangmangu Baru’ towards
developing them as plant growth-promoting bacteria in improving garlic quality
and productivity. The isolation of endophytic bacteria was performed from
garlic leaves and roots during the vegetative phase. A total of 20 endophytic
bacteria isolated from garlic leaves and roots were characterised by their
morphology and tested for their ability to fix nitrogen (N), produce auxin
(IAA), and solubilise phosphate. All the isolates were able to fix nitrogen,
with 15 of the isolates found to be Gram-positive having bacilli cell shape.
Furthermore, 8 isolates were shown to produce auxin, while 10 isolates were
able to solubilise phosphate, with both groups originating from the root
sample. Five potential isolates were identified as nitrogen-fixing,
IAA-producing, and phosphate-solubilising, with the highest values of IAA
concentration and the Phosphate Solubilisation Index (PSI) being 10.71 ppm and
2.00, respectively. These 5 potential endophytic bacterial isolates were
genetically identified as similar to Lysobacter tolerans, Enterobacter cloacae, Enterobacter huaxiensis, Klebsiella oxytoca,
and Enterobacter hormaechei. The potential
endophytic bacterial isolates hold promise for developing garlic plant
growth-promoting bacteria (PGPB) to enhance garlic production both
qualitatively and quantitatively.
Keywords: Allium
sativum; endophytic bacteria; IAA; nitrogen; phosphate
Abstrak
Permintaan yang tinggi untuk bawang putih (Allium sativum L.) membawa kepada usaha untuk meningkatkan produktivitinya. Bakteria endofit bawang putih didapati mempunyai keupayaan untuk meningkatkan pertumbuhan dan melindungi tumbuhan bawang putih daripada penyakit, tanpa kesan berbahaya pada perumah. Objektif kajian ini adalah untuk memencilkan dan mencirikan bakteria endofit yang terdapat dalam bawang putih tempatan Indonesia var ‘Tawangmangu Baru’ bagi membangunkannya sebagai bakteria penggalak pertumbuhan tumbuhan dalam meningkatkan kualiti dan produktiviti bawang putih. Pemencilan bakteria endofit dilakukan daripada daun dan akar bawang putih semasa fasa vegetatif. Sebanyak 20 bakteria endofit yang dipencilkan daripada daun dan akar bawang putih telah dicirikan morfologinya dan diuji kebolehannya untuk mengikat nitrogen (N), menghasilkan auksin (IAA) dan melarutkan fosfat. Semua pencilan dapat mengikat nitrogen dengan 15 pencilan didapati sebagai Gram-positif yang mempunyai bentuk rod. Tambahan pula, 8 pencilan telah menunjukkan keupayaan menghasilkan auksin, manakala 10 pencilan berupaya melarutkan fosfat, yang kesemuanya berasal daripada sampel akar. Sebanyak 5 pencilan berpotensi dikenal pasti sebagai pengikat nitrogen, penghasil IAA dan pelarut fosfat, dengan nilai kepekatan IAA dan IPF Indeks Pelarutan Fosfat (IPF) tertinggi, masing-masing ialah 10.71 ppm dan
2.00. Kesemua 5 pencilan bakteria endofit yang berpotensi ini dikenal pasti secara genetik menyamai Lysobacter tolerans, Enterobacter cloacae, Enterobacter huaxiensis, Klebsiella oxytoca dan Enterobacter hormaechei. Pencilan bakteria endofit yang berpotensi merupakan prospek dalam membangunkan bakteria penggalak pertumbuhan bawang putih untuk meningkatkan pengeluaran bawang putih secara kualitatif dan kuantitatif.
Kata kunci: Allium sativum; bakteria endofit; fosfat; IAA;
nitrogen
REFERENCES
Azizah, H., Rahajeng,
S.M. & Jatmiko, Y.D. 2020. Isolation and
screening of phosphate and potassium solubilizing endophytic bacteria in maize
(Zea mays L.). Journal of
Experimental Life Science 10(3): 165-170.
Bhattacharjee, A., Dubey, S. & Sharma,
S. 2023. “Next-Generation Bioformulations” for plant growth promotion and
stress mitigation: A promising approach for sustainable agriculture. Journal
of Plant Growth Regulation 42(10): 6741-6759.
BPS-Statistics Indonesia. 2023. Vegetable
Crop Production (Indonesian).
https://www.bps.go.id/indicator/55/61/1/produksi-tanaman-sayuran.html
Costa Júnior, P.S.P., Cardoso, F.P.,
Martins, A.D., Buttrós, V.H.T., Pasqual, M., Dias,
D.R., Schwan, R.F. & Dória, J. 2020. Endophytic
bacteria of garlic roots promote growth of micropropagated meristems. Microbiological Research 241: 126585.
Ek-Ramos, M.J., Gomez-Flores, R.,
Orozco-Flores, A.A., Rodríguez-Padilla, C., González-Ochoa, G. & Tamez-Guerra, P. 2019. Bioactive products from plant-endophytic Gram-positive bacteria. Frontiers
in Microbiology 10: 463.
Ekowati, C.N., Mirani, M., Handayani,
K. & Agustrina, R. 2021. Detection of nitrogenase producing bacteria
from the soil of Liwa botanical garden. Jurnal Ilmu Biologi Eksperimen dan Keanekaragaman Hayati 8: 53-58.
Fadeev, E., Cardozo-Mino, M.G., Rapp, J.Z., Bienhold, C., Salter, I., Salman-Carvalho, V., Molari, M., Tegetmeyer, H.E.,
Buttigieg, P.L. & Boetius, A. 2021. Comparison of
two 16S rRNA primers (V3–V4 and V4–V5) for studies of arctic microbial
communities. Frontiers in Microbiology 12: 637526.
Gómez Expósito,
R., Postma, J., Raaijmakers, J.M. & De Bruijn, I. 2015. Diversity and activity of Lysobacter species from disease suppressive soils. Frontiers
in Microbiology 6: 166241.
Gordon, S.A. & Weber, R.P. 1951.
Colorimetric estimation of indoleacetic acid. Plant Physiology 26(1):
192-195.
Gunstone, T., Cornelisse,
T., Klein, K., Dubey, A. & Donley, N. 2021. Pesticides and soil
invertebrates: A hazard assessment. Frontiers in Environmental Science 9: 643847.
Halimah, D., Munif, A.
& Giyanto, G. 2016. The potential of Ochrobactrum intermedium - C939A31, Klebsiella oxytoca - C939A32, Bacillus subtilis -
I308A32 isolated from coffee plant for controlling root lesion nematode Pratylenchus coffeae. Jurnal Fitopatologi Indonesia 12(2): 62-68.
Jha, P.N. & Kumar, A. 2007. Endophytic
colonization of Typha australis by a plant
growth-promoting bacterium Klebsiella oxytoca strain GR-3. Journal of Applied Microbiology 103(4): 1311-1320.
Ji, C., Liu, Z., Hao, L., Song, X., Wang,
C., Liu, Y., Li, H., Li, C., Gao, Q. & Liu, X. 2020. Effects of Enterobacter
cloacae HG-1 on the nitrogen-fixing community structure of wheat
rhizosphere soil and on salt tolerance. Frontiers in Plant Science 11: 510947.
Kaga, H., Mano, H., Tanaka, F., Watanabe, A.,
Kaneko, S. & Morisaki, H. 2009. Rice seeds as
sources of endophytic bacteria. Microbes and Environments 24: 154-162.
Khianngam, S., Techakriengkrai,
T., Raksasiri, B.V., Kanjanamaneesathian,
M. & Tanasupawat, S. 2013. Isolation and screening of endophytic
bacteria for hydrolytic enzymes from plant in mangrove forest at Pranburi, Prachuap Khiri Khan,
Thailand. In Endophytes for Plant Protection: The State of the Art. Proc.
5th Int. Symp. Plant Protect. Plant Health Europe,
edited by Schneider, C., Leifert, C. & Feldmann,
F. Berlin: Deutsche Phytomedizinische Gesellschaft.
pp. 279-284.
Koskey, G., Mburu, S.W., Awino,
R., Njeru, E.M. & Maingi, J.M. 2021. Potential
use of beneficial microorganisms for soil amelioration, phytopathogen
biocontrol, and sustainable crop production in smallholder agroecosystems. Frontiers
in Sustainable Food Systems 5: 606308.
Lin, H., Liu, C., Peng, Z., Tan, B., Wang,
K. & Liu, Z. 2022. Distribution pattern of endophytic bacteria and fungi in
tea plants. Frontiers in Microbiology 13: 872034.
Matos, A.D.M., Gomes, I.C.P., Nietsche, S. & Xavier, A.A. 2017. Phosphate
solubilization by endophytic bacteria isolated from banana trees. The Annals
of the Brazilian Academy of Sciences 89(4): 2945-2954.
Ministry of Agriculture. 2023. Strategic
Food Commodity Analysis in 2023 (Indonesian). Jakarta: Ministry of Agriculture.
Pande, A., Pandey, P., Mehra,
S., Singh, M. & Kaushik, S. 2017. Phenotypic and genotypic characterization
of phosphate solubilizing bacteria and their efficiency on the growth of maize. Journal of Genetic Engineering and Biotechnology 15(2): 379-391.
Purwaningsih, R., Dewi, W.
& Pujiasmanto, B. 2019. Indigenous
phosphate-solubilizing bacteria enhance germination in deteriorated rice seed. Bulgarian
Journal of Agricultural Science 25(3): 486-493.
Rana, K.L., Kour,
D., Kaur, T., Devi, R., Yadav, A.N., Yadav, N., Dhaliwal, H.S. & Saxena,
A.K. 2020. Endophytic microbes: Biodiversity, plant growth-promoting mechanisms
and potential applications for agricultural sustainability. Antonie van Leeuwenhoek 113(8): 1075-1107.
Ryan, R.P., Germaine, K., Franks, A., Ryan,
D.J. & Dowling, D.N. 2008. Bacterial endophytes: Recent developments and
applications. FEMS Microbiology Letters 278(1): 1-9.
Shofiyah, L., Sudadi, Dewi, W.S. & Cahyani, V.R.
2023. Endophytic phosphate solubilization and potential nitrogen-fixing
bacteria in the leaf and root of rice planted on the conventional wetland. IOP
Conference Series: Earth and Environmental Science 1162(1): 012007.
Silitonga, D.M, Priyani,
N. & Nurwahyuni, I. 2008. Isolation and potential
testing of phosphate-solubilizing bacteria and IAA (Indole Acetic Acid)
hormone-producing bacteria on soybean (Glicine max L.) growth on yellow soil (Indonesian). Medan: Sumatera Utara
University.
Singh, H., Khar,
A. & Verma, P. 2021. Induced mutagenesis for genetic improvement of Allium
genetic resources: A comprehensive review. Genetic Resources and Crop
Evolution 68(7): 2669-2690.
Smith, A. & Hussey, M. 2005. Gram stain
protocols. American Society for Microbiology 1(14): 113-144.
Srivastav, A.L., Patel, N., Rani, L.,
Kumar, P., Maddodi, I.D.B.S. & Chaudhary, V.K.
2024. Sustainable options for fertilizer management in agriculture to prevent
water contamination: A review. Environment, Development and Sustainability 26:
8303-8327.
Walia, A., Guleria, S.,
Chauhan, A. & Mehta, P. 2017. Endophytic bacteria: Role in phosphate
solubilization. In Endophytes: Crop Productivity and Protection. Sustainable
Development and Biodiversity, edited by Maheshwari, D. & Annapurna, K.
Springer, Cham.
Wang, Z., Solanki, M.K., Pang, F., Singh,
R.K., Yang, L-T., Li, Y-R., Li, H-B., Zhu, K. & Xing, Y-X. 2017.
Identification and efficiency of a nitrogen-fixing endophytic actinobacterial
strain from sugarcane. Sugar Tech 19: 492-500.
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