Functional role of an endophytic Bacillus amyloliquefaciens in enhancing growth and disease protection of invasive English ivy (Hedera helix L.)
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We hypothesize that invasive English ivy (Hedera helix) harbors endophytic microbes that promote plant growth and survival. To evaluate this hypothesis, we examined endophytic bacteria in English ivy and evaluated effects on the host plant.
Endophytic bacteria were isolated from multiple populations of English ivy in New Brunswick, NJ. Bacteria were identified as a single species Bacillus amyloliquefaciens. One strain of B. amyloliquefaciens, strain C6c, was characterized for indoleacetic acid (IAA) production, secretion of hydrolytic enzymes, phosphate solubilization, and antibiosis against pathogens. PCR was used to amplify lipopeptide genes and their secretion into culture media was detected by MALDI-TOF mass spectrometry. Capability to promote growth of English ivy was evaluated in greenhouse experiments. The capacity of C6c to protect plants from disease was evaluated by exposing B+ (bacterium inoculated) and B− (non-inoculated) plants to the necrotrophic pathogen Alternaria tenuissima.
B. amyloliquefaciens C6c systemically colonized leaves, petioles, and seeds of English ivy. C6c synthesized IAA and inhibited plant pathogens. MALDI-TOF mass spectrometry analysis revealed secretion of antifungal lipopeptides surfactin, iturin, bacillomycin, and fengycin. C6c promoted the growth of English ivy in low and high soil nitrogen conditions. This endophytic bacterium efficiently controlled disease caused by Alternaria tenuissima.
This study suggests that B. amyloliquefaciens plays an important role in enhancing growth and disease protection of English ivy.
KeywordsLipopeptide Biological control Plant growth promotion Invasive plants
The Federal University of Mato Grosso (UFMT), Department of Plant Biology and Pathology of Rutgers University; The Brazilian National Council for Scientific and Technological Development (CNPq) for Post Doctoral Fellowship; International Institute of Science and Technology in Wetlands (INAU); and Sr. Qiang Chen for confocal microscopy assistance were acknowledged. The authors are also grateful to the support from the John E. and Christina C. Craighead Foundation, USDA-NIFA Multistate Project W3147, and the New Jersey Agricultural Experiment Station. Any use of trade, product or firm names is for descriptive purposes only and does not imply endorsement by the U.S. Government. This article is Contribution 1957 of the USGS Great Lakes Science Center.
- Aravind R, Eapen SJ, Kumar A, Dinu A, Ramana KV (2010) Screening of endophytic bacteria and evaluation of selected isolates for suppression of burrowing nematode (Radopholus similis Thorne) using three varieties of black pepper (Piper nigrum L.). Crop Prot 29:318–324. doi: 10.1016/j.cropro.2009.12.005 CrossRefGoogle Scholar
- Bacon CW, White JF (2000) Microbial endophytes. Marcel Dekker, New YorkGoogle Scholar
- Beltran-Garcia MJ, White Jr, JF, Prado FM, Prieto KR, Yamaguchi LF, Torres MS, Di Mascio P (2014) Nitrogen acquisition in Agave tequilana from degradation of endophytic bacteria. Sci Rep 4. doi: 10.1038/srep06938
- Berg G, Krechel A, Ditz M, Sikora RA, Ulrich A, Hallmann J (2005a) Endophytic and ectophytic potato-associated bacterial communities differ in structure and antagonistic function against plant pathogenic fungi. FEMS Microbiol Ecol 51:215–229. doi: 10.1016/j.femsec.2004.08.006 PubMedCrossRefGoogle Scholar
- Borriss R, Chen XH, Rueckert C, Blom J, Becker A, Baumgarth B, Klenk HP (2011) Relationship of Bacillus amyloliquefaciens clades associated with strains DSM 7T and FZB42T: a proposal for Bacillus amyloliquefaciens subsp. amyloliquefaciens subsp. nov. and Bacillus amyloliquefaciens subsp. plantarum subsp. nov. based on complete genome sequence comparisons. Int J Syst Evol Microbiol 61:1786–1801. doi: 10.1099/ijs.0.023267-0 PubMedCrossRefGoogle Scholar
- Chung S, Kong H, Buyer JS, Lakshman DK, Lydon J, Kim SD, Roberts DP (2008) Isolation and partial characterization of Bacillus subtilis ME488 for suppression of soilborne pathogens of cucumber and pepper. Appl Microbiol Biotechnol 80:115–123. doi: 10.1007/s00253-008-1520-4 PubMedCrossRefGoogle Scholar
- Clapham AR, Tutin TG, Moore DM (1990) Flora of the British isles. CUP ArchiveGoogle Scholar
- Dabundo R, Lehmann MF, Treibergs L, Tobias CR, Altabet MA, Moisander PH, Granger J (2014) The contamination of commercial 15N2 gas stocks with 15N-labeled nitrate and ammonium and consequences for nitrogen fixation measurements. PLoS ONE 9:e110335. doi: 10.1371/journal.pone.0110335 PubMedPubMedCentralCrossRefGoogle Scholar
- Ellenberg H (1988) Vegetation ecology of central Europe. Cambridge University Press, CambridgeGoogle Scholar
- González‐Sánchez MÁ, Pérez‐Jiménez RM, Pliego C, Ramos C, De Vicente A, Cazorla FM (2010) Biocontrol bacteria selected by a direct plant protection strategy against avocado white root rot show antagonism as a prevalent trait. J Appl Microbiol 109:65–78. doi: 10.1111/j.1365-2672.2009.04628.x PubMedGoogle Scholar
- Grime JP, Hodgson JG, Hunt R (1988) Comparative plant ecology. A functional approach to common British species. Unwin Hyman Ltd, LondonGoogle Scholar
- Jourdan E, Henry G, Duby F, Dommes J, Barthélemy JP, Thonart P, Ongena MARC (2009) Insights into the defense-related events occurring in plant cells following perception of surfactin-type lipopeptide from Bacillus subtilis. Plant Microbe Interact 22:456–468. doi: 10.1094/MPMI-22-4-0456 CrossRefGoogle Scholar
- Kowalski KP, Bacon C, Bickford W, Braun H, Clay K, Leduc-Lapierre M, Lillard E, McCormick MK, Nelson E, Torres M, White J, Wilcox DA (2015) Advancing the science of microbial symbiosis to support invasive species management: a case study on Phragmites in the Great Lakes. Front Microbiol 6:95. doi: 10.3389/fmicb.2015.00095 PubMedPubMedCentralCrossRefGoogle Scholar
- Kumaresan V, Suryanarayanan TS (2002) Endophyte assemblages in young, mature and senescent leaves of Rhizophora apiculata: evidence for the role of endophytes in mangrove litter degradation. Fungal Divers 9:81–91Google Scholar
- Leelasuphakul W, Hemmanee P, Chuenchitt S (2008) Growth inhibitory properties of Bacillus subtilis strains and their metabolites against the green mold pathogen (Penicillium digitatum Sacc.) of citrus fruit. Postharvest Biol Technol 48:113–121. doi: 10.1016/j.postharvbio.2007.09.024 CrossRefGoogle Scholar
- Montanez A, Blanco AR, Barlocco C, Beracochea M, Sicardi M (2012) Characterization of cultivable putative endophytic plant growth promoting bacteria associated with maize cultivars (Zea mays L.) and their inoculation effects in vitro. Appl Soil Ecol 58:21–28. doi: 10.1016/j.apsoil.2012.02.009 CrossRefGoogle Scholar
- Norris JR, Chapman HM (1968) Classification of Azotobacter. In: Gibbs BM, Shapton DA (eds) Identification methods for microbiologists. Academic Press, New York, pp 19–27Google Scholar
- Panchal H, Ingle S (2011) Isolation and characterization of endophytes from the root of the medicinal plant Chlorophytum borivilianum (Safed musli). J Adv Dev Res 2:205–209Google Scholar
- Rajagopal R, Suryanarayanan TS (2000) Isolation of endophytic fungi from leaves of neem (Azadirachta indica). Curr Sci 78:1375–1378Google Scholar
- Romero D, de Vicente A, Rakotoaly RH, Dufour SE, Veening JW, Arrebola E, Pérez-García A (2007) The iturin and fengycin families of lipopeptides are key factors in antagonism of Bacillus subtilis toward Podosphaera fusca. Mol Plant Microbe Interact 20:430–440. doi: 10.1094/MPMI-20-4-0430 PubMedCrossRefGoogle Scholar
- Singh N, Pandey P, Dubey RC, Maheshwari DK (2008) Biological control of root rot fungus Macrophomina phaseolina and growth enhancement of Pinus roxburghii (Sarg.) by rhizosphere competent Bacillus subtilis BN1. World J Microbiol Biotechnol 24:1669–1679. doi: 10.1007/s11274-008-9680-z CrossRefGoogle Scholar
- Souto GI, Correa OS, Montecchia MS, Kerber NL, Pucheu NL, Bachur M, Garcia AF (2004) Genetic and functional characterization of a Bacillus sp. strain excreting surfactin and antifungal metabolites partially identified as iturin‐like compounds. J Appl Microbiol 97:1247–1256. doi: 10.1111/j.1365-2672.2004.02408.x PubMedCrossRefGoogle Scholar
- Stevenson FJ, Cole MA (1999) Cycles of soil: carbon, nitrogen phosphorus, sulfur, and micronutrients. Wiley, New YorkGoogle Scholar
- Suryanarayanan TS, Vijaykrishna D (2001) Fungal endophytes of aerial roots of Ficus benghalensis. Fungal Divers 8:155–161Google Scholar
- Sylvester-Bradley R, Asakawa N, La Torraca S, Magalhães FMM, Oliveira L, Pereira RM (1982) Levantamento quantitativo de microrganismos solubilizadores de fosfatos na rizosfera de gramíneas e leguminosas forrageiras na Amazônia. Acta Amazon 12:15–22Google Scholar
- Westbrook R (1998) Invasive plants: weeds of the global garden. Brooklyn Botanical Garden, New YorkGoogle Scholar
- Whitehouse R (2006) Which ivy? Metula Books, LondonGoogle Scholar
- Xu M, Sheng J, Chen L, Men Y, Gan L, Guo S, Shen L (2014) Bacterial community compositions of tomato (Lycopersicum esculentum Mill.) seeds and plant growth promoting activity of ACC deaminase producing Bacillus subtilis (HYT-12-1) on tomato seedlings. World J Microbiol Biotechnol 30:835–845. doi: 10.1007/s11274-013-1486-y PubMedCrossRefGoogle Scholar
- Yuan J, Ruan Y, Wang B, Zhang J, Waseem R, Huang Q, Shen Q (2013) Plant growth-promoting rhizobacteria strain Bacillus amyloliquefaciens NJN-6-enriched bio-organic fertilizer suppressed Fusarium wilt and promoted the growth of banana plants. J Agric Food Chem 61:3774–3780. doi: 10.1021/jf400038z PubMedCrossRefGoogle Scholar
- Zhao P, Quan C, Jin L, Wang L, Wang J, Fan S (2013) Effects of critical medium components on the production of antifungal lipopeptides from Bacillus amyloliquefaciens Q-426 exhibiting excellent biosurfactant properties. World J Microbiol Biotechnol 29:401–409. doi: 10.1007/s11274-012-1180-5 PubMedCrossRefGoogle Scholar