Holaphyllamine, a steroid, is able to induce defense responses in Arabidopsis thaliana and increases resistance against bacterial infection
- 405 Downloads
A chemical screen of plant-derived compounds identified holaphyllamine, a steroid, able to trigger defense responses in Arabidopsis thaliana and improve resistance against the pathogenic bacterium Pseudomonas syringae pv tomato DC3000.
A chemical screen of 1600 plant-derived compounds was conducted and allowed the identification of a steroid able to activate defense responses in A. thaliana at a concentration of 1 µM without altering growth. The identified compound is holaphyllamine (HPA) whose chemical structure is similar to steroid pregnanes of mammals. Our data show that HPA, which is not constitutively present in A. thaliana, is able to trigger the formation of reactive oxygen species, deposition of callose and expression of several pathogenesis-related genes of the salicylic and jasmonic acid pathways. In addition, the results show that pre-treatment of A. thaliana seedlings with HPA before infection with the pathogenic bacterium Pseudomonas syringae pv tomato DC3000 results in a significant reduction of symptoms (i.e., reduction of bacterial colonies). Using A. thaliana mutants, we have found that the activation of defense responses by HPA does not depend on BRI1/BAK1 receptor kinases. Finally, a structure/function study reveals that the minimal structure required for activity is a 5-pregnen-20-one steroid with an equatorial nucleophilic group in C-3. Together, these findings demonstrate that HPA can activate defense responses that lead to improved resistance against bacterial infection in A. thaliana.
KeywordsCallose Chemical screen Elicitor Holaphyllamine PATHOGENESIS-RELATED Plant defense Pseudomonas syringae pv tomato DC3000 ROS Steroid
BRASSINOSTEROID INSENSITIVE1-ASSOCIATED RECEPTOR-LIKE KINASE1
Pseudomonas syringae pv tomato DC3000
We are grateful to Pr. Jianming Li (University of Michigan, Department of Molecular, Cellular, and Developmental Biology, Ann Arbor, MI, USA) for the kind gift of the bri1-9 mutant.
Compliance with ethical standards
This work was supported by the research network “Végétal-Agronomie-Sol-Innovations” (VASI) de Haute-Normandie and the Fonds Européen de Développement Régional (FEDER) in the ALTERAGRO project and the French ministry of Research and Higher Education. The post-doctoral scholarship of AZ was funded by FEDER. Cell imaging, microplate fluorescence detection and qRT-PCR analyses were performed at PRIMACEN (http://www.primacen.fr), the Cell Imaging Platform of Normandy, IRIB, Normandie Univ, UniRouen, 76821 Mont-Saint-Aignan, France.
Conflict of interest
The authors declare that they have no conflict of interest.
- Abramoff MD, Magalhaes PJ, Ram SJ (2004) Image processing with ImageJ. Biophot Int 11:36–43Google Scholar
- Albrecht C, Boutrot F, Segonzac C, Schwessinger B, Gimenez-Ibanez S, Chinchilla D, Rathjen JP, de Vries SC, Zipfel C (2012) Brassinosteroids inhibit pathogen-associated molecular pattern-triggered immune signaling independent of the receptor kinase BAK1. Proc Natl Acad Sci USA 109:303–308. doi: 10.1073/pnas.1109921108 CrossRefPubMedGoogle Scholar
- Bellien J, Remy-Jouet I, Iacob M, Blot E, Mercier A, Lucas D, Dreano Y, Gutierrez L, Donnadieu N, Thuillez C, Joannides R (2012) Impaired role of epoxyeicosatrienoic acids in the regulation of basal conduit artery diameter during essential hypertension. Hypertension 60:1415–1421. doi: 10.1161/HYPERTENSIONAHA.112.201087 CrossRefPubMedGoogle Scholar
- Buell CR, Joardar V, Lindeberg M, Selengut J, Paulsen IT, Gwinn ML, Dodson RJ, Deboy RT, Durkin AS, Kolonay JF et al (2003) The complete genome sequence of the Arabidopsis and tomato pathogen P. syringae pv. tomato DC3000. Proc Natl Acad Sci USA 100:10181–10186. doi: 10.1073/pnas.1731982100 CrossRefPubMedPubMedCentralGoogle Scholar
- Do Rego JL, Seong JY, Burel D, Leprince J, Luu-The V, Tsutsui K, Tonon MK, Pelletier G, Vaudry H (2009) Neurosteroid biosynthesis: enzymatic pathways and neuroendocrine regulation by neurotransmitters and neuropeptides. Front Neuroendocrinol 30:259–301. doi: 10.1016/j.yfrne.2009.05.006 CrossRefPubMedGoogle Scholar
- Gauthier A, Trouvelot S, Kelloniemi J, Frettinger P, Wendehenne D, Daire X, Joubert JM, Ferrarini A, Delledonne M, Flors V, Poinssot B (2014) The sulfated laminarin triggers a stress transcriptome before priming the SA- and ROS-dependent defenses during grapevine’s induced resistance against Plasmopara viticola. PLoS One 9:e88145. doi: 10.1371/journal.pone.0088145 CrossRefPubMedPubMedCentralGoogle Scholar
- Janot MM, Cave A, Goutarel R (1960) Steroid alkaloids. Holaphyllamine and holamine, alkaloids from Holarrhena floribunda (G. Don) Dur. and Schinz. CR Hebd Séances Acad Sci 251:559–561Google Scholar
- Laquitaine L, Gomès E, François J, Marchive C, Pascal S, Hamdi S, Atanassova R, Delrot S, Coutos-Thévenot P (2006) Molecular basis of ergosterol-induced protection of grape against Botrytis cinerea: induction of type I LTP promoter activity, WRKY, and stilbene synthase gene expression. Mol Plant-Microbe Interact 19:1103–1112. doi: 10.1094/MPMI-19-1103 CrossRefPubMedGoogle Scholar
- Ménard R, Alban S, de Ruffray P, Jamois F, Franz G, Fritig B, Yvin JC, Kauffmann S (2004) β-1,3 glucan sulfate, but not β-1,3 glucan, induces the salicylic acid signaling pathway in tobacco and Arabidopsis. Plant Cell 16:3020–3032. doi: 10.1105/tpc.104.024968 CrossRefPubMedPubMedCentralGoogle Scholar
- Noutoshi Y, Okazaki M, Kida T, Nishina Y, Morishita Y, Ogawa T, Hideyuki S, Daisuke S, Yusuke J, Atsushi H, Yuji K, Ken S (2012) Novel plant immune-priming compounds identified via high-throughput chemical screening target salicylic acid glucosyltransferases in Arabidopsis. Plant Cell 24:3795–3804. doi: 10.1105/tpc.112.098343 CrossRefPubMedPubMedCentralGoogle Scholar
- Plancot B, Santaella C, Jaber R, Kiefer-Meyer MC, Follet-Gueye ML, Leprince J, Gattin I, Souc C, Driouich A, Vicré-Gibouin M (2013) Deciphering the responses of root border-like cells of Arabidopsis thaliana and Linum usitatissimum to pathogen-derived elicitors. Plant Physiol 163:1584–1597. doi: 10.1104/pp.113.222356 CrossRefPubMedPubMedCentralGoogle Scholar
- Postel S, Küfner I, Beuter C, Mazzotta S, Schwedt A, Borlotti A, Halter T, Kemmerling B, Nürnberger T (2010) The multifunctional leucine-rich repeat receptor kinase BAK1 is implicated in Arabidopsis development and immunity. Eur J Cell Biol 89:169–174. doi: 10.1016/j.ejcb.2009.11.001 CrossRefPubMedGoogle Scholar
- Sanchez L, Courteaux B, Hubert J, Kauffman S, Renault JH, Clement C, Baillieul F, Dorey S (2012) Rhamnolipids elicit defense responses and induce disease resistance against biotrophic, hemibiotrophic and necrotrophic pathogens that require different signalling pathways in Arabidopsis thaliana and highlight a central role for salicylic acid. Plant Physiol 160:1630–1641. doi: 10.1104/pp.112.201913 CrossRefPubMedPubMedCentralGoogle Scholar
- Schreiber KJ, Nasmith CG, Allard GC, Singh J, Subramaniam R, Desveaux D (2011) High-throughput chemical screening in Arabidopsis thaliana identifies small molecules that reduce Fusarium head blight disease in wheat. Mol Plant Microbe Interact 24:640–648. doi: 10.1094/MPMI-09-10-0210 CrossRefPubMedGoogle Scholar
- Shi CL, Park HB, Lee JK, Ryu S, Ryu CM (2010) Inhibition of primary roots and stimulation of lateral root development in Arabidopsis thaliana by the rhizobacterium Serratia marcescens 90–166 is through both auxin-dependent and independent signaling pathways. Mol Cells 29:251–258. doi: 10.1007/s10059-010-0032-0 CrossRefPubMedGoogle Scholar