Abstract
Main conclusion
Exogenous SA treatment at appropriate concentrations promotes adventitious root formation in cucumber hypocotyls, via competitive inhibiting the IAA-Asp synthetase activity of CsGH3.5, and increasing the local free IAA level.
Abstract
Adventitious root formation is critical for the cutting propagation of horticultural plants. Indole-3-acetic acid (IAA) has been shown to play a central role in regulating this process, while for salicylic acid (SA), its exact effects and regulatory mechanism have not been elucidated. In this study, we showed that exogenous SA treatment at the concentrations of both 50 and 100 µM promoted adventitious root formation at the base of the hypocotyl of cucumber seedlings. At these concentrations, SA could induce the expression of CYCLIN and Cyclin-dependent Kinase (CDK) genes during adventitious rooting. IAA was shown to be involved in SA-induced adventitious root formation in cucumber hypocotyls. Exposure to exogenous SA led to a slight increase in the free IAA content, and pre-treatment with the auxin transport inhibitor 1-naphthylphthalamic acid (NPA) almost completely abolished the inducible effects of SA on adventitious root number. SA-induced IAA accumulation was also associated with the enhanced expression of Gretchen Hagen3.5 (CsGH3.5). The in vitro enzymatic assay indicated that CsGH3.5 has both IAA- and SA-amido synthetase activity and prefers aspartate (Asp) as the amino acid conjugate. The Asp concentration dictated the functional activity of CsGH3.5 on IAA. Both affinity and catalytic efficiency (Kcat/Km) increased when the Asp concentration increased from 0.3 to 1 mM. In contrast, CsGH3.5 showed equal catalytic efficiency for SA at low and high Asp concentrations. Furthermore, SA functioned as a competitive inhibitor of the IAA-Asp synthetase activity of CsGH3.5. During adventitious formation, SA application indeed repressed the IAA-Asp levels in the rooting zone. These data show that SA plays an inducible role in adventitious root formation in cucumber through competitive inhibition of the auxin conjugation enzyme CsGH3.5. SA reduces the IAA conjugate levels, thereby increasing the local free IAA level and ultimately enhancing adventitious root formation.
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Abbreviations
- ARL1:
-
Adventitious rootless1
- Asp:
-
Aspartate
- dpe:
-
Day post-excision
- GH3:
-
Gretchen Hagen3
- GRXC9:
-
Glutaredoxin C9
- GST:
-
Glutathione S-transferase
- NPA:
-
1-Naphthylphthalamic acid
- PAT:
-
Polar auxin transport
- PIN:
-
PIN-FORMED
- PR1-1a:
-
Pathogenesis-related 1-1a
- SA:
-
Salicylic acid
- SAUR:
-
Small auxin-up RNA
- YUC:
-
YUCCA
References
Agullo-Anton MA, Ferrandez-Ayela A, Fernandez-Garcia N, Nicolas C, Albacete A, Perez-Alfocea F, Sanchez-Bravo J, Perez-Perez JM, Acosta M (2014) Early steps of adventitious rooting: morphology, hormonal profiling and carbohydrate turnover in carnation stem cuttings. Physiol Plant 150:446–462. https://doi.org/10.1111/ppl.12114
Armengot L, Marquès-Bueno MM, Soria-Garcia A, Müller M, Munné-Bosch S, Martínez MC (2014) Functional interplay between protein kinase CK2 and salicylic acid sustains PIN transcriptional expression and root development. Plant J 78:411–423. https://doi.org/10.1111/tpj.12481
Bellini C, Pacurar DI, Perrone I (2014) Adventitious roots and lateral roots: similarities and differences. Annu Rev Plant Biol 65:639–666. https://doi.org/10.1146/annurev-arplant-050213-035645
Blanco F, Garretón V, Frey N, Dominguez C, Pérez-Acle T, Van der Straeten D, Jordana X, Holuigue L (2005) Identification of NPR1-dependent and independent genes early induced by salicylic acid treatment in Arabidopsis. Plant Mol Biol 59:927–944. https://doi.org/10.1007/s11103-005-2227-x
Böttcher C, Boss PK, Davies C (2011) Acyl substrate preferences of an IAA-amido synthetase account for variations in grape (Vitis vinifera L.) berry ripening caused by different auxinic compounds indicating the importance of auxin conjugation in plant development. J Exp Bot 62:4267–4280. https://doi.org/10.1093/jxb/err134
Böttcher C, Dennis EG, Booker GW, Polyak SW, Boss PK, Davies C (2012) A novel tool for studying auxin-metabolism: the inhibition of grapevine indole-3-acetic acid-amido synthetases by a reaction intermediate analogue. PLoS ONE 7:e37632. https://doi.org/10.1371/journal.pone.0037632
Bourne DJ, Barrow KD, Milborrow BV (1991) Salicyloylaspartate as an endogenous component in the leaves of Phaseolus vulgaris. Phytochemistry 30:4041–4044. https://doi.org/10.1016/0031-9422(91)83462-T
Cano A, Sánchez-García AB, Albacete A, González-Bayón R, Justamante MS, Ibáñez S, Acosta M, Pérez-Pérez JM (2018) Enhanced conjugation of auxin by GH3 enzymes leads to poor adventitious rooting in carnation stem cuttings. Front Plant Sci 9:566. https://doi.org/10.3389/fpls.2018.00566
Chen Y, Shen H, Wang M, Li Q, He Z (2013) Salicyloyl-aspartate synthesized by the acetyl-amido synthetase GH3.5 is a potential activator of plant immunity in Arabidopsis. Acta Biochim Biophys Sin 45:827–836. https://doi.org/10.1093/abbs/gmt078
Chen L, Tong J, Xiao L, Ruan Y, Liu J, Zeng M, Huang H, Wang JW, Xu L (2016) YUCCA-mediated auxin biogenesis is required for cell fate transition occurring during de novo root organogenesis in Arabidopsis. J Exp Bot 67:4273–4284. https://doi.org/10.1093/jxb/erw213
De Klerk GJ, Guan HY, Huisman P, Marinova S (2011) Effects of phenolic compounds on adventitious root formation and oxidative decarboxylation of applied indoleacetic acid in Malus ‘Jork 9’. Plant Growth Regul 63:175–185. https://doi.org/10.1007/s10725-010-9555-9
Ding P, Ding Y (2020) Stories of salicylic acid: a plant defense hormone. Trends Plant Sci 25:549–565. https://doi.org/10.1016/j.tplants.2020.01.004
Ding X, Cao Y, Huang L, Zhao J, Xu C, Li X, Wang S (2008) Activation of the indole-3-acetic acid-amido synthetase GH3-8 suppresses expansin expression and promotes salicylate- and jasmonate-independent basal immunity in rice. Plant Cell 20:228–240. https://doi.org/10.1105/tpc.107.055657
Dong CJ, Cao N, Li L, Shang QM (2016a) Quantitative proteomic profiling of early and late responses to salicylic acid in cucumber leaves. PLoS ONE 11:e0161395. https://doi.org/10.1371/journal.pone.0161395
Dong C, Cao N, Wang L, Zhang H, Wang H, Tai L, Shang Q (2016b) Regulatory roles of cotyledon-generated auxin in adventitious root formation on the hypocotyls of cucumber seedlings. Acta Hortic Sin 43:1929–1940. https://doi.org/10.16420/j.issn.0513-353x.2016-0184
Druege U, Hilo A, Pérez-Pérez JM, Klopotek Y, Acosta M, Shahinnia F, Zerche S, Franken F, Hajirezaei MR (2019) Molecular and physiological control of adventitious rooting in cuttings: phytohormone action meets resource allocation. Ann Bot 123:929–949. https://doi.org/10.1093/aob/mcy234
Ehmann A (1977) The van Urk-Salkowski reagent—A sensitive and specific chromogenic reagent for silica gel thin-layer chromatographic detection and identification of indole derivatives. J Chromatogr 132:267–276. https://doi.org/10.1016/s0021-9673(00)89300-0
Ermel FF, Vizoso S, Charpentier JP, Jay-Allemand C, Catesson AM, Couée I (2000) Mechanisms of primordium formation during adventitious root development from walnut cotyledon explants. Planta 211:563–574. https://doi.org/10.1007/s004250000314
Fu ZQ, Dong X (2013) Systemic acquired resistance: turning local infection into global defense. Annu Rev Plant Biol 64:839–863. https://doi.org/10.1146/annurev-arplant-042811-105606
Guan L, Tayengwa R, Cheng Z, Peer WA, Murphy AS, Zhao M (2019) Auxin regulates adventitious root formation in tomato cuttings. BMC Plant Biol 19:435. https://doi.org/10.1186/s12870-019-2002-9
Gutierrez L, Mongelard G, Floková K, Pacurar DI, Novák O, Staswick P, Kowalczyk M, Păcurar M, Demailly H, Geiss G, Bellini C (2012) Auxin controls Arabidopsis adventitious root initiation by regulating jasmonic acid homeostasis. Plant Cell 24:2515–2527. https://doi.org/10.1105/tpc.112.099119
Herrera-Vásquez A, Carvallo L, Blanco F, Tobar M, Villarroel-Candia E, Vicente-Carbajosa J, Salinas P, Holuigue L (2015) Transcriptional control of glutaredoxin GRXC9 expression by a salicylic acid-dependent and NPR1-independent pathway in Arabidopsis. Plant Mol Biol Report 33:624–637. https://doi.org/10.1007/s11105-014-0782-5
Jain M, Kaur N, Tyagi AK, Khurana JP (2006) Structure and expression analysis of early auxin-responsive Aux/IAA gene family in rice (Oryza sativa). Funct Integr Genomics 6:36–46. https://doi.org/10.1007/s10142-005-0005-0
Jasik J, De Klerk GJ (1997) Anatomical and ultrastructural examination of adventitious root formation in stem slices of apple. Biol Plant 39:79–90. https://doi.org/10.1023/A:1000313207486
Kling GJ, Myer MM (1983) Effects of phenolic compounds and indoleacetic acid on adventitious root initiation in cuttings of Phaseolus aureus, Acer saccharinum, and Acer griseum. HortScience 18:352–354. https://doi.org/10.1007/s10725-010-9555-9
Koo YM, Heo AY, Choi HW (2020) Salicylic acid as a safe plant protector and growth regulator. Plant Pathol J 36:1–10. https://doi.org/10.5423/PPJ.RW.12.2019.0295
Kora D, Bhattacharjee S (2020) The interaction of reactive oxygen species and antioxidants at the metabolic interface in salicylic acid-induced adventitious root formation in mung bean [Vigna radiata (L.) R. Wilczek]. J Plant Physiol 248:153152. https://doi.org/10.1016/j.jplph.2020.153152
Lakehal A, Bellini C (2018) Control of adventitious root formation: insights into synergistic and antagonistic hormonal interactions. Physiol Plant 165:90–100. https://doi.org/10.1111/ppl.12823
Lakehal A, Dob A, Novák O, Bellini C (2019) A DAO1-mediated circuit controls auxin and jasmonate crosstalk robustness during adventitious root initiation in Arabidopsis. Int J Mol Sci 20:4428. https://doi.org/10.3390/ijms20184428
Liu H, Wang S, Yu X, Yu J, He X, Zhang S, Shou H, Wu P (2005) ARL1, a LOB-domain protein required for adventitious root formation in rice. Plant J 43:47–56. https://doi.org/10.1111/j.1365-313X.2005.02434.x
Ljung K (2013) Auxin metabolism and homeostasis during plant development. Development 140:943–950. https://doi.org/10.1242/dev.086363
Mackelprang R, Okrent RA, Wildermuth MC (2017) Preference of Arabidopsis thaliana GH3.5 acyl amido synthetase for growth versus defense hormone acyl substrates is dictated by concentration of amino acid substrate aspartate. Phytochemistry 143:19–28. https://doi.org/10.1016/j.phytochem.2017.07.001
Mignolli F, Mariotti L, Picciarelli P, Vidoz ML (2017) Differential auxin transport and accumulation in the stem base lead to profuse adventitious root primordia formation in the aerial roots (aer) mutant of tomato (Solanum lycopersicum L.). J Plant Physiol 213:55–65. https://doi.org/10.1016/j.jplph.2017.02.010
Miura K, Tada Y (2014) Regulation of water, salinity, and cold stress responses by salicylic acid. Front Plant Sci 5:4. https://doi.org/10.3389/fpls.2014.00004
Nobuta K, Okrent RA, Stoutemyer M, Rodibaugh N, Kempema L, Wildermuth MC, Innes RW (2007) The GH3 acyl adenylase family member PBS3 regulates salicylic acid-dependent defense responses in Arabidopsis. Plant Physiol 144:1144–1156. https://doi.org/10.1104/pp.107.097691
Okrent RA, Brooks MD, Wildermuth MC (2009) Arabidopsis GH3.12 (PBS3) conjugates amino acids to 4-substituted benzoates and is inhibited by salicylate. J Biol Chem 284:9742–9754. https://doi.org/10.1074/jbc.M806662200
Pacurar DI, Perrone I, Bellini C (2014) Auxin is a central player in the hormone cross-talks that control adventitious rooting. Physiol Plant 151:83–96. https://doi.org/10.1111/ppl.12171
Park JE, Park JY, Kim YS, Staswick PE, Jeon J, Yun J, Kim SY, Kim J, Lee YH, Park CM (2007) GH3-mediated auxin homeostasis links growth regulation with stress adaptation response in Arabidopsis. J Biol Chem 282:10036–10046. https://doi.org/10.1074/jbc.M610524200
Pasternak T, Groot EP, Kazantsev FV, Teale W, Omelyanchuk N, Kovrizhnykh V, Palme K, Mironova VV (2019) Salicylic acid affects root meristem patterning via auxin distribution in a concentration-dependent manner. Plant Physiol 180:1725–1739. https://doi.org/10.1104/pp.19.00130
Qi X, Li Q, Ma X, Qian C, Wang H, Ren N, Shen C, Huang S, Xu X, Xu Q, Chen X (2019) Waterlogging-induced adventitious root formation in cucumber is regulated by ethylene and auxin through reactive oxygen species signaling. Plant Cell Environ 42:1458–1470. https://doi.org/10.1111/pce.13504
Riov J, Yang SF (1989) Enhancement of adventitious root formation in mung bean cuttings by 3,5-dihalo-4-hydroxybenzoic acids and 2,4-dinitrophenol. Plant Growth Regul 8:277–281. https://doi.org/10.1007/BF00025397
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425. https://doi.org/10.1093/oxfordjournals.molbev.a040454
Shakirova FM, Sakhabutdinova AR, Bezrukova MV, Fatkhutdinova RA, Fatkhutdinova DR (2003) Changes in the hormonal status of wheat seedlings induced by salicylic acid and salinity. Plant Sci 164:317–322. https://doi.org/10.1016/S0168-9452(02)00415-6
Shang QM, Li L, Dong CJ (2012) Multiple tandem duplication of the phenylalanine ammonia-lyase genes in Cucumis sativus L. Planta 236:1093–1105. https://doi.org/10.1007/s00425-012-1659-1
Shen J, Zhang Y, Ge D, Wang Z, Song W, Gu R, Che G, Cheng Z, Liu R, Zhang X (2019) CsBRC1 inhibits axillary bud outgrowth by directly repressing the auxin efflux carrier CsPIN3 in cucumber. Proc Natl Acad Sci USA 116:17105–17114. https://doi.org/10.1073/pnas.1907968116
Sorin C, Negroni L, Balliau T, Corti H, Jacquemot MP, Davanture M, Sandberg G, Zivy M, Bellini C (2006) Proteomic analysis of different mutant genotypes of Arabidopsis led to the identification of 11 proteins correlating with adventitious root development. Plant Physiol 140:349–364. https://doi.org/10.1104/pp.105.067868
Staswick PE, Tiryaki I, Rowe M (2002) Jasmonate response locus JAR1 and several related Arabidopsis genes encode enzymes of the firefly luciferase superfamily that show activity on jasmonic, salicylic, and indole-3-acetic acids in an assay for adenylation. Plant Cell 14:1405–1415. https://doi.org/10.1105/tpc.000885
Staswick PE, Serban B, Rowe M, Tiryaki I, Maldonado MT, Maldonado MC, Suza W (2005) Characterization of an Arabidopsis enzyme family that conjugates amino acids to indole-3-acetic acid. Plant Cell 17:616–627. https://doi.org/10.1105/tpc.104.026690
Steffens B, Rasmussen A (2016) The physiology of adventitious roots. Plant Physiol 170:603–617. https://doi.org/10.1104/pp.15.01360
Terol J, Domingo C, Talon M (2006) The GH3 family in plants: genome wide analysis in rice and evolutionary history based on EST analysis. Gene 371:279–290. https://doi.org/10.1016/j.gene.2005.12.014
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The ClustalX windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882. https://doi.org/10.1093/nar/25.24.4876
Vlot AC, Dempsey DMA, Klessig DF (2009) Salicylic acid, a multifaceted hormone to combat disease. Annu Rev Phytopathol 47:177–206. https://doi.org/10.1146/annurev.phyto.050908.135202
Wang J, Wu B, Lu K, Wei Q, Qian J, Chen Y, Fang Z (2019) The amino acid permease 5 (OsAAP5) regulates tiller number and grain yield in rice. Plant Physiol 180:1031–1045. https://doi.org/10.1104/pp.19.00034
Westfall CS, Herrmann J, Chen Q, Wang S, Jez JM (2010) Modulating plant hormones by enzyme action: the GH3 family of acyl acid amido synthetases. Plant Signal Behav 5:1607–1612. https://doi.org/10.4161/psb.5.12.13941
Westfall CS, Zubieta C, Herrmann J, Kapp U, Nanao MH, Jez JM (2012) Structural basis for pre-receptor modulation of plant hormones by GH3 family proteins. Science 336:1708–1711. https://doi.org/10.1126/science.1221863
Westfall CS, Sherp AM, Zubieta C, Alvarez S, Schraft E, Marcellin R, Ramirez L, Jez JM (2016) Arabidopsis thaliana GH3.5 acyl acid amido synthetase mediates metabolic crosstalk in auxin and salicylic acid homeostasis. Proc Natl Acad Sci USA 113:13917–13922. https://doi.org/10.1073/pnas.1612635113
Wu J, Liu S, Guan X, Chen L, He Y, Wang J, Lu G (2014) Genome-wide identification and transcriptional profiling analysis of auxin response-related gene families in cucumber. BMC Res Notes 7:218. https://doi.org/10.1186/1756-0500-7-218
Xu L (2018) De novo root regeneration from leaf explants: wounding, auxin, and cell fate transition. Curr Opin Plant Biol 41:39–45. https://doi.org/10.1016/j.pbi.2017.08.004
Xu L, Zhao H, Ruan W, Deng M, Wang F, Peng J, Luo J, Chen Z, Yi K (2017) Abnormal inflorescence meristem1 functions in salicylic acid biosynthesis to maintain proper reactive oxygen species levels for root meristem activity in rice. Plant Cell 29:560–574. https://doi.org/10.1105/tpc.16.00665
Xu X, Ji J, Xu Q, Qi X, Weng Y, Chen X (2018) The major-effect quantitative trait locus CsARN6.1 encodes an AAA ATPase domain-containing protein that is associated with waterlogging stress tolerance by promoting adventitious root formation. Plant J 93:917–930. https://doi.org/10.1111/tpj.13819
Yan S, Che G, Ding L, Chen Z, Liu X, Wang H, Zhao W, Ning K, Zhao J, Tesfamichael K, Wang Q, Zhang X (2016) Different cucumber CsYUC genes regulate response to abiotic stresses and flower development. Sci Rep 6:20760. https://doi.org/10.1038/srep20760
Yang W, Zhu C, Ma X, Li G, Gan L, Ng D, Xia K (2013) Hydrogen peroxide is a second messenger in the salicylic acid-triggered adventitious rooting process in mung bean seedlings. PLoS ONE 8:e84580. https://doi.org/10.1371/journal.pone.0084580
Yang H, Klopotek Y, Hajirezaei MR, Zerche S, Franken P, Druege U (2019) Role of auxin homeostasis and response in nitrogen limitation and dark stimulation of adventitious root formation in petunia cuttings. Ann Bot 124:1053–1066. https://doi.org/10.1093/aob/mcz095
Zhang Y, Li X (2019) Salicylic acid: biosynthesis, perception, and contributions to plant immunity. Curr Opin Plant Biol 50:29–36. https://doi.org/10.1016/j.pbi.2019.02.004
Zhang Z, Li Q, Li Z, Staswick PE, Wang M, Zhu Y, He Z (2007) Dual regulation role of GH3.5 in salicylic acid and auxin signaling during Arabidopsis-Pseudomonas syringae interaction. Plant Physiol 145:450–464. https://doi.org/10.1104/pp.107.106021
Zhang SW, Li CH, Cao J, Zhang YC, Zhang SQ, Xia YF, Sun DY, Sun Y (2009) Altered architecture and enhanced drought tolerance in rice via the downregulation of indole-3-acetic acid by TLD1/OsGH3.13 activation. Plant Physiol 151:1889–1901. https://doi.org/10.1104/pp.109.146803
Zhang Q, Huber H, Beljaars SJM, Birnbaum D, de Best S, de Kroon H, Visser EJW (2017) Benefits of flooding-induced aquatic adventitious roots depend on the duration of submergence: linking plant performance to root functioning. Ann Bot 120:171–180. https://doi.org/10.1093/aob/mcx049
Zhao Y (2018) Essential roles of local auxin biosynthesis in plant development and in adaptation to environmental changes. Annu Rev Plant Biol 69:417–435. https://doi.org/10.1146/annurev-arplant-042817-040226
Zhao X, Wang J, Yuan J, Wang XL, Zhao QP, Kong PT, Zhang X (2015) NITRIC OXIDE-ASSOCIATED PROTEIN1 (AtNOA1) is essential for salicylic acid-induced root waving in Arabidopsis thaliana. New Phytol 207:211–224. https://doi.org/10.1111/nph.13327
Zheng Z, Guo Y, Novák O, Chen W, Ljung K, Noel JP, Chory J (2016) Local auxin metabolism regulates environment-induced hypocotyl elongation. Nat Plants 2:16025. https://doi.org/10.1038/nplants.2016.25
Zhu X, Galili G (2003) Increased lysine synthesis coupled with a knockout of its catabolism synergistically boosts lysine content and also transregulates the metabolism of other amino acids in Arabidopsis seeds. Plant Cell 15:845–853. https://doi.org/10.1105/tpc.009647
Acknowledgements
This work was supported by grants from the National Key R&D Program of China (No. 2019YFD1000305), Central Public-interest Scientific Institution Basal Research Fund (No. IVF-BRF2019013), the China Agriculture Research System (CARS-25), and the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences (CAASASTIP-IVFCAAS).
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Dong, CJ., Liu, XY., Xie, LL. et al. Salicylic acid regulates adventitious root formation via competitive inhibition of the auxin conjugation enzyme CsGH3.5 in cucumber hypocotyls. Planta 252, 75 (2020). https://doi.org/10.1007/s00425-020-03467-2
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DOI: https://doi.org/10.1007/s00425-020-03467-2