Volatile metabolites of endophytic Klebsiella aerogenes from Zingiber zerumbet rhizome and its antagonistic effect on soft rot causative Pythium myriotylum
DOI:
https://doi.org/10.25081/josac.2020.v29.i2.6553Keywords:
antagonistic, endophyte, soft-rot, volatile organic compoundsAbstract
Rhizomes of Zingiber zerumbet collected from their natural habitat and reported earlier to have high zerumbone content were selected for isolation of endophytes. Biochemical and molecular characterization using 16S rRNA sequencing of the endophytes identified the isolates as belonging to genus Klebsiella, Pantoea and Enterobacter. Isolate designated ZzKSD8 identified as K. aerogenes yielded maximal antagonistic activities against P. myriotylum determined as 83.5% ± 0.77. Volatile metabolites produced by ZzKSD8 caused absolute impairment of P. myriotylum hyphal growth compared to control (23.9 ± 0.37 cm). Volatile metabolites were extracted from 48 hour grown stationary phase ZzKSD8 cultures using absolute methanol, ethyl acetate and ethanol (60% v/v). GC-MS metabolite profiling detected alkanes and fatty acid methyl esters as the predominant constituents in the solvent extracts. Major constituents included methyl palmitate (31.37%), methyl stearate (18.57%) and cyclopropaneoctanoic acid, 2-hexyl-, methyl ester (17.05%) in methanol extract; alkanes like tetratetracontane (13.18%) and 2-methyloctacosane (12.10%) in ethyl acetate extracts and 4,22-sigmastadiene-3-one as major metabolite (22.51%) and stigmast-5-en-3-ol, (3.beta) (17.40%) in ethanol extract. Identified metabolites reported to modulate defense strategies in plants against phytopathogens makes ZzKSD8 a potential candidate for development of biological alternatives to control soil-borne soft-rot disease.
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References
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Aleklett K, Leff JW, Fierer N & Hart M 2015 Wild plant species growing closely connected in a subalpine meadow host distinct root-associated bacterial communities. Peer J. 3: e804.
Baby S, Dan M, Thaha A R M, Johnson A J, Kurup R, Balakrishnapillai P & Lim C K 2009 High content of zerumbone in volatile oils of Zingiber zerumbet from southern India and Malaysia. Flav. Frag. J. 24: 301–308.
Banik A, Mukhopadhaya S K & Dangar T K 2016 Characterization of N2-fixing plant growth promoting endophytic and epiphytic bacterial community of Indian cultivated and wild rice (Oryza spp.) genotypes. Planta 243: 799–812.
Chaparro J M, Sheflin A M, Manter D K & Vivanco J M 2012 Manipulating the soil microbiome to increase soil health and plant fertility. Biol. Fert. Soils 48: 489–499.
Chien T Y, Chen L G, Lee C J, Lee F Y & Wang C C 2008 Anti-inflammatory constituents of Zingiber zerumbet. Food Chem. 110: 584–589.
Dong Y, Chelius M K, Brisse S, Kozyrovska N, Kovtunovych G, Podschun R & Triplett E W 2003 Comparisons between two Klebsiella: The plant endophyte K. pneumoniae 342 and a clinical isolate, K. pneumoniae MGH78578. Symbiosis 35: 247–259.
El Arab H G D, Vilich V & Sikora R A 2001 The use of phospholipid fatty acids (PL-FA) in the determination of rhizosphere specific microbial communities (RSMC) of two wheat cultivars. Plant Soil 228: 291–297.
Felsenstein J 1981 Evolutionary trees from DNA sequences: a maximum likelihood approach. J. Mol. Evol. 17: 368–376.
Gaiero J R, McCall C A, Thompson K A, Day N J, Best A S & Dunfield K E 2013 Inside the root microbiome: bacterial root endophytes and plant growth promotion. Am. J. Bot. 100: 1738–1750.
Hoover T R 2000 Control of nitrogen fixation genes in Klebsiella pneumonia. In: Triplett EW (Ed.) Prokaryotic Nitrogen Fixation: A model system for te analysis of a biological process (pp. 131–147). Horizon Scientific Press, Norfolk, UK
Jia M, Chen L, Xin H L, Zheng C-J, Rahman K, Han T & Qin L-P 2016 A friendly relationship between endophytic fungi and medicinal plants: A systematic review. Front Microbiol. 7: 906.
Jishma P, Hussain N, Chellappan R, Rajendran R, Mathew J & Radhakrishnan E K 2017 Strain-specific variation in plant growth promoting volatile organic compounds production by five different Pseudomonas spp. as confirmed by response of Vigna radiata seedlings. J. Appl. Microbiol. 123: 204–216.
Keerthi D, Geethu C, Nair R A, Pillai P 2014 Metabolic profiling of Zingiber zerumbet following Pythium myriotylum infection: Investigations on the defensive role of the principal secondary metabolite, Zerumbone. Appl. Biochem. Biotechnol. 172: 2593–2603.
Keerthi D, Aswati N R & Prasath D 2016 Molecular phylogenetics and anti-Pythium activity of endophytes from rhizomes of wild ginger congener, Zingiber zerumbet Smith. World J. Microbiol. Biotechnol. 32: 41.
Kumar S, Stecher G, Li M, Knyaz C & Tamura K 2018 MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 35: 1547–1549.
Lu Q, Liu T, Wang N, Dou Z, Wang K & Zuo Y 2020 Nematicidal effect of Methyl Palmitate and Methyl Stearate against Meloidogyne incognita in Bananas. J. Agric. Food Chem. 68: 24: 6502–6510.
Macedo-Raygoza G M, Valdez-Salas B, Prado F M, Prieto K R, Yamaguchi L F, Kato M J, Canto-Canché B B, Carrillo-Beltrán M, Di Mascio P, White J F & Beltrán-García M J 2019 Enterobacter cloacae, an endophyte that establishes a nutrient-transfer symbiosis with banana plants and protects against the Black Sigatoka pathogen. Front. Microbiol. 10: 804.
Martinez L, Caballero-Mellaod J, Orozco J & Martinez-Romero E 2003 Diazotrophic bacteria associated with banana (Musa spp.). Plant Soil 257: 35–47.
Mendes R, Garbeva P & Raaijmakers J M 2013 The rhizosphere microbiome: significance of plant beneficial, plant pathogenic, and human pathogenic microorganisms. FEMS Microbiol. Reviews 379: 634–663.
Mishra A, Chauhan P S, Chaudhry V, Tripathi M & Nautiyal C S 2011 Rhizosphere competent Pantoea agglomerans enhances maize (Zea mays) and chickpea (Cicer arietinum L.) growth, without altering the rhizosphere functional diversity. Antonie Van Leeuwenhoek. 100: 405–13.
Nair R A & Thomas G 2013 Molecular characterization of ZzR1 resistance gene from Zingiber zerumbet with potential for imparting Pythium aphanidermatum resistance in ginger. Gene 516: 58–65.
Nigris S, Baldan E & Tondello A 2018 Biocontrol traits of Bacillus licheniformis GL174, a culturable endophyte of Vitis vinifera cv. Glera. BMC Microbiol. 18: 133.
Overbeek L V & Elsas J D V 2008 Effects of plant genotype and growth stage on the structure of bacterial communities associated with potato (Solanum tuberosum L.). FEMS Microbiol. Ecol. 64: 283–296.
Raaijmakers J M, Paulitz T C, Steinberg C, Alabouvette C & Moënne-Loccoz Y 2009 The rhizosphere: a playground and battlefield for soilborne pathogens and beneficial microorganisms. Plant Soil 321: 341–361.
Ruiz B, Chávez A, Forero A, García-Huante Y, Romero A, Sánchez M, Rocha D, Sánchez B, Rodriguez-Sanoja R, Sanchez S & Langley E 2010 Production of microbial secondary metabolites: Regulation by the carbon source. Crit. Rev. Microbiol. 36: 146–167.
Ryan R P, Germaine K, Franks A, Ryan D J & Dowling D N 2007 Bacterial endophytes: recent developments and applications. FEMS Microbiol. Lett. 278: 1–9.
Santoyo G, Moreno-Hagelsieb G, Orozco-Mosqueda M C & Glick B R 2016 Plant growth-promoting bacterial endophytes. Microbiol. Res. 183: 92–99.
Shubin L, Juan H, RenChao Z, ShiRu X & YuanXiao J 2014 Fungal endophytes of Alpinia officinarum rhizomes: insights on diversity and variation across growth years, growth sites, and the inner active chemical concentration. PLoS One 9: e115289.
Strobel G & Daisy B 2003 Bioprospecting for microbial endophytes and their natural products. Microbiol. Mol. Biol. Rev. 67: 491–502.
Thompson J D, Higgins D G & Gibson T J 1994 CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22: 4673–4680.
Torres A R, Araújo W L, Cursino L, Hungria M, Plotegher F, Mostasso F L &f Azevedo J L 2008 Diversity of endophytic enterobacteria associated with different host plants. J. Microbiol. 46: 373–379.
Tyc O, Song C, Dickschat J S, Vos M & Garbeva P 2017 The ecological role of volatile and soluble secondary metabolites produced by soil bacteria. Trends Microbiol. 25: 280–292.
William S, Feil H & Copeland A 2012 Bacterial genomic DNA isolation using CTAB. Sigma 50: 6876.
Zhang Y, Kang X, Liu H, Liu Y, Li Y, Yu X, Zhao K, Gu Y, Xu K, Chen C & Chen Q 2018 Endophytes isolated from ginger rhizome exhibit growth promoting potential for Zea mays. Arch. Agron. Soil Sci. 64: 1302–1314.
Published
30-12-2020
How to Cite
Harsha, K., & Aswati Nair, R. (2020). Volatile metabolites of endophytic Klebsiella aerogenes from Zingiber zerumbet rhizome and its antagonistic effect on soft rot causative Pythium myriotylum. Journal of Spices and Aromatic Crops, 29(2), 122–132. https://doi.org/10.25081/josac.2020.v29.i2.6553
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