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Callus, shoot and hairy root formation in vitro as affected by the sensitivity to auxin and ethylene in tomato mutants

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Abstract

We analyzed the impact of ethylene and auxin disturbances on callus, shoots and Agrobacterium rhizogenes-induced hairy root formation in tomato (Solanum lycopersicum L.). The auxin low-sensitivity dgt mutation showed little hairy root initiation, whereas the ethylene low-sensitivity Nr mutation did not differ from the control Micro-Tom cultivar. Micro-Tom and dgt hairy roots containing auxin sensitivity/biosynthesis rol and aux genes formed prominent callus onto media supplemented with cytokinin. Under the same conditions, Nr hairy roots did not form callus. Double mutants combining Rg1, a mutation conferring elevated shoot formation capacity, with either dgt or Nr produced explants that formed shoots with little callus proliferation. The presence of rol + aux genes in Rg1 hairy roots prevented shoot formation. Taken together, the results suggest that although ethylene does not affect hairy root induction, as auxin does, it may be necessary for auxin-induced callus formation in tomato. Moreover, excess auxin prevents shoot formation in Rg1.

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Abbreviations

MT:

Micro-Tom

Rg1 :

Regeneration locus 1

dgt :

diageotropica

Nr :

Never ripe

rol :

root loci

pRi :

Root inductor plasmid

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

References

  • Aloni R, Wolf A, Feigenbaum P, Avni A, Klee HJ (1998) The Never-ripe mutant provides evidence that tumor-induced ethylene controls the morphogenesis of Agrobacterium tumefaciens-induced crown galls on tomato plants. Plant Physiol 177:841–849

    Article  Google Scholar 

  • Auer CA, Cohen JD, Laloue M, Cooke TJ (1992) Comparison of benzyladenine metabolism in two Petunia lines differing in shoot organogenesis. Plant Physiol 98:1035–1041

    Article  PubMed  CAS  Google Scholar 

  • Baumann K, De Paolis A, Costantino P, Gualberti G (1999) The DNA biding site of the Dof protein NtBBF1 is essential for tissue-specific and auxin-regulated expression of the rolB oncogene in plants. Plant Cell 11:323–333

    Article  PubMed  CAS  Google Scholar 

  • Bishop GJ, Nomura T, Yokota T, Harrison K, Noguchi T, Fujioka S, Takatsuto S, Jones JDG, Kamiya Y (1999) The tomato DWARF enzyme catalyses C-6 oxidation in brassinosteroid biosynthesis. Proc Natl Acad Sci USA 96:1761–1766

    Article  PubMed  CAS  Google Scholar 

  • Boiten H, Azmi A, Dillen W, De Schepper S, Debergh P, Gerats T, Van Onckelen H, Prinsen H (2004) The Rg-1 encoded regeneration capacity of tomato is not related to an altered cytokinin homeostasis. New Phytol 161:761–771

    Article  CAS  Google Scholar 

  • Cardarelli M, Spanó L, Mariotti D, Mauro ML, Van Sluys MA, Costantino P (1987) The role of auxin in hairy root induction. Mol Gen Genet 208:457–463

    Article  CAS  Google Scholar 

  • Cary A, Uttamchandani SJ, Smets R, Van Onckelen HA, Howell SHH (2001) Arabidopsis mutants with increased organ regeneration in tissue culture are more competent to respond to hormonal signals. Planta 213:700–707

    Article  PubMed  CAS  Google Scholar 

  • Centeno ML, Rodríguez A, Feito I, Fernández B (1996) Relationship between endogenous auxin and cytokinin levels and morphogenic responses in Actinidia deliciosa tissue cultures. Plant Cell Rep 16:58–62

    Article  CAS  Google Scholar 

  • Chatfield SP, Raizada MN (2008) Ethylene and shoot regeneration: hookless1 modulates de novo shoot organogenesis in Arabidopsis thaliana. Plant Cell Rep 27:655–666

    Article  PubMed  CAS  Google Scholar 

  • Chilton MD, Tepfer DA, Petit A, David C, Casse Delbart F, Tempé J (1982) Agrobacterium rhizogenes inserts T-DNA into genomes of the host plant root cells. Nature 295:432–434

    Article  CAS  Google Scholar 

  • Chriqui D, Guivarch A, Dewitte W, Prinsen E, van Onkelen H (1996) Rol genes and root initiation and development. Plant Soil 187:47–55

    Article  CAS  Google Scholar 

  • Clark DG, Gubrium EK, Barrett JE, Nell TA, Klee HJ (1999) Root formation in the ethylene-insensitive plants. Plant Physiol 121:53–59

    Article  PubMed  CAS  Google Scholar 

  • Delbarre A, Muller P, Imhoff V, Barbier-Brygoo H, Maurel C, Leblanc N, Perrot-Rechenmann C, Guern J (1994) The rolB gene of Agrobacterium rhizogenes does not increase the auxin sensitivity of tobacco protoplasts by modifying the intracellular auxin concentration. Plant Physiol 105:563–569

    PubMed  CAS  Google Scholar 

  • Faiss M, Strnad M, Redig P, Dolezal K, Hanus J, Van Onckelen H, Schmülling T (1996) Chemically induced expression of the rolC-encoded beta-glucosidase in transgenic tobacco plants and analysis of cytokinin metabolism: rolC does not hydrolyze endogenous cytokinin glucosides in planta. Plant J 10:33–46

    Article  CAS  Google Scholar 

  • Fulton TM, Chunwongse J, Tanksley SD (1995) Microprep protocol for extraction of DNA from tomato and other herbaceous plants. Plant Mol Biol Rep 13:207–209

    Article  CAS  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirement of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  PubMed  CAS  Google Scholar 

  • Goda H, Sawa S, Asami T, Fujioka S, Shimada Y, Yoshida S (2004) Comprehensive comparison of auxin-regulated and brassinosteroid-regulated genes in Arabidopsis. Plant Physiol 134:1555–1573

    Article  PubMed  CAS  Google Scholar 

  • Hardtkea C, Dorceya E, Osmonta KS, Sibout R (2007) Phytohormone collaboration: zooming in on auxin–brassinosteroid interactions. Trend Cell Biol 17:485–492

    Article  CAS  Google Scholar 

  • Hicks GR, Rayle DL, Lomax TL (1989) The diageotropica mutant of tomato lacks high specific activity auxin binding sites. Science 245:52–54

    Article  PubMed  CAS  Google Scholar 

  • Kepczynski J, McKersie D, Brown DCW (1992) Requirement of ethylene for growth of callus and somatic embryogenesis in Medicago sativa L. J Exp Bot 43:1199–1202

    Article  CAS  Google Scholar 

  • Klee HJ, Estelle M (1991) Molecular genetic approaches to plant hormone biology. Annu Rev Pant Physiol Plant Mol Biol 42:529–551

    Article  CAS  Google Scholar 

  • Koka CV, Cerny RE, Gardner RG, Noguchi T, Fujioka S, Takatsuto S, Yoshida S, Clouse SD (2000) A putative role for the tomato genes DUMPY and CURL-3 in brassinosteroid biosynthesis and response. Plant Physiol 122:85–98

    Article  PubMed  CAS  Google Scholar 

  • Koornneef M, Bade J, Hanhart C, Horsman K, Schel J, Soppe W, Verkerk R, Zabel P (1993) Characterization and mapping of a gene controlling shoot regeneration in tomato. Plant J 3:131–141

    Article  CAS  Google Scholar 

  • Koornneef M, Alonso-Blanco C, Peeters AJM (1997) Genetic approaches in plant physiology. New Phytol 137:1–8

    Article  Google Scholar 

  • Kumar PP, Lakshmanan P, Thorpe TA (1998) Regulation of morphogenesis in plant tissue culture by ethylene. In Vitro Cell Dev Biol Plant 34:94–103

    Article  CAS  Google Scholar 

  • Lakshmanan P, Ng SK, Loh CS, Goh CJ (1997) Auxin, cytokinin and ethylene differentially regulate specific developmental states associated with shoot bud morphogenesis in leaf tissues of mangosteen (Garcinia mangostana L.) cultured in vitro. Plant Cell Physiol 38:59–64

    CAS  Google Scholar 

  • Lanahan MB, Yan H-C, Giovannoni JJ, Klee HJ (1994) The Never ripe mutation blocks ethylene perception in tomato. Plant Cell 6:521–530

    Article  PubMed  CAS  Google Scholar 

  • Lima JE, Carvalho RF, Neto AT, Figueira A, Peres LEP (2004) Micro-MsK: a tomato genotype with miniature size, short life cycle, and improved in vitro shoot regeneration. Plant Sci 167:753–757

    Article  CAS  Google Scholar 

  • Martí E, Gisbert C, Bishop GJ, Dixon MS, Garcia-Martinez JL (2006) Genetic and physiological characterization of tomato cv. Micro-Tom. J Exp Bot 57:2037–2047

    Article  PubMed  CAS  Google Scholar 

  • Meissner R, Jacobson Y, Melamed S, Levyatuv S, Shalev G, Ashri A, Elkind Y, Levy A (1997) A new model system for tomato genetics. Plant J 12:1465–1472

    Article  CAS  Google Scholar 

  • Moritz T, Schmülling T (1998) The gibberellin content of rolA transgenic tobacco plants is specifically altered. J Plant Physiol 153:774–776

    CAS  Google Scholar 

  • Mouchel CF, Osmont KS, Hardtke CS (2006) BRX mediates feedback between brassinosteroid level and auxin signaling in root growth. Nature 443:458–461

    Article  PubMed  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Oh K, Ivanchenko MG, White TJ, Lomax TL (2006) The diageotropica gene of tomato encodes a cyclophilin: a novel player in auxin signaling. Planta 224:133–144

    Article  PubMed  CAS  Google Scholar 

  • Paciorek T, Friml J (2006) Auxin signaling. J Cell Sci 119:1199–1202

    Article  PubMed  CAS  Google Scholar 

  • Peres LEP, Kerbauy GB (1999) High cytokinin accumulation following root tip excision changes the endogenous auxin to cytokinin ratio during root-to-shoot conversion in Catasetum fimbriatum Lindl. (Orchidaceae). Plant Cell Rep 18:1002–1006

    Article  CAS  Google Scholar 

  • Peres LEP, Amar S, Kerbauy GB, Salatino A, Zaffari GR, Mercier H (1999) Effects of auxin, cytokinin and ethylene treatments on the endogenous ethylene and auxin-to-cytokinin ratio related to direct root tip conversion of Catasetum fimbriatum Lindl. (Orchidaceae) into buds. J Plant Physiol 155:551–555

    CAS  Google Scholar 

  • Peres LEP, Morgante PG, Vechi C, Kraus JE, Van Sluys M-A (2001) Shoot regeneration capacity from roots and transgenic hairy roots of different tomato cultivars and wild related species. Plant Cell Tissue Organ Cult 65:37–44

    Article  CAS  Google Scholar 

  • Petit A, Tempé J (1978) Isolation of Agrobacterium Ti-plasmid regulatory mutants. Mol Gen Genet 167:147–155

    Article  CAS  Google Scholar 

  • Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:2002–2007

    Article  Google Scholar 

  • Ricker AJ (1930) Studies on infections hairy roots of nursery apple trees. J Agric Res 41:438–446

    Google Scholar 

  • Sarul P, Vlahova M, Ivanova A, Atanassov A (1995) Direct shoot formation in spontaneously occurring root pseudonodules of alfalfa (Medicago sativa L.). In Vitro Cell Dev Biol 31:21–25

    Article  Google Scholar 

  • Scott JW, Harbaugh BK (1989) Micro-Tom, a miniature dwarf tomato. Florida Agricultural Sciences, University of Florida, Circular S-370:1–6

  • Serrani JC, Sanjuan R, Ruiz-Rivero O, Fos M, Garcia-Martinez JL (2007) Gibberellin regulation of fruit set and growth in tomato. Plant Physiol 145:246–257

    Article  PubMed  CAS  Google Scholar 

  • Shen WH, Petit A, Guern J, Tempé J (1988) Hairy roots are more sensitive to auxin than normal roots. Proc Natl Acad Sci USA 85:3417–3421

    Article  PubMed  CAS  Google Scholar 

  • Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp Soc Exp Biol 11:118–131

    Google Scholar 

  • Stevens MA, Rick CM (1986) Genetic and breeding. In: Atherton JG, Rudich J (eds) The tomato crop: a scientific basis for improvement. Chapman and Hall, London, pp 35–109

    Google Scholar 

  • Sugiyama M (1999) Organogenesis in vitro. Curr Opin Plant Biol 2:61–64

    Article  PubMed  CAS  Google Scholar 

  • Tanimoto M, Roberts K, Dolan L (1995) Ethylene is a positive regulator of root hair development in Arabidopsis thaliana. Plant J 6:943–948

    Google Scholar 

  • Tieman DM, Ciardi JA, Taylor MG, Klee HJ (2001) Members of the tomato LeEIL (EIN3-like) gene family are functionally redundant and regulate ethylene responses throughout plant development. Plant J 26:47–58

    Article  PubMed  CAS  Google Scholar 

  • Vasil IK (2008) A history of plant biotechnology: from the cell theory of Schleiden and Schwann to biotech crops. Plant Cell Rep 27:1423–1440

    Article  PubMed  CAS  Google Scholar 

  • Wang H, Jones B, Li ZG, Frasse P, Delalande C, Regad F, Chaabouni S, Latche A, Pech JC, Bouzayen M (2005) The tomato Aux/IAA transcription factor IAA9 is involved in fruit development and leaf morphogenesis. Plant Cell 17:2676–2692

    Article  PubMed  CAS  Google Scholar 

  • Wilkinson JQ, Lanahan MB, Yen HC, Giovannoni JJ, Klee HJ (1995) An ethylene-inducible component of signal-transduction encoded by Never-ripe. Science 270:1807–1809

    Article  PubMed  CAS  Google Scholar 

  • Yamada T, Palm CJ, Brooks B, Kosuge T (1985) Nucleotide sequences of the Pseudomonas savastanoi indoleacetic acid genes show homology with Agrobacterium tumefaciens T-DNA. Proc Natl Acad Sci USA 82:6522–6526

    Article  PubMed  CAS  Google Scholar 

  • Yoshimatsu K, Shimomura K (1994) Plant regeneration on cultured root segments of Cephalis ipecacuanha A. Richard. Plant Cell Rep 14:98–101

    Article  CAS  Google Scholar 

  • Zsögön A, Lambais MR, Benedito VA, Figueira AVO, Peres LEP (2008) Reduced arbuscular mycorrhizal colonization in tomato ethylene mutants. Sci Agric 65:259–267

    Article  Google Scholar 

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Acknowledgments

The following are acknowledged: FAPESP (grant 02/00329-8 and fellowship 03/03940-2) and CNPq (grant 475494/03-2 and fellowship 308075/03-0) for financial support.

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Author notes

  1. Joni Esrom Lima

    Present address: Institut für Pflanzenernährung, Universität Hohenheim, 70593, Stuttgart, Germany

  2. Vagner Augusto Benedito

    Present address: Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Pky, Ardmore, OK, 73401, USA

Authors and Affiliations

  1. Department of Biological Sciences (LCB), Escola Superior de Agricultura “Luiz de Queiroz” (ESALQ), Universidade de São Paulo (USP), Av. Pádua Dias, 11, CP 09, Piracicaba, SP, 13418-900, Brazil

    Lázaro Eustáquio Pereira Peres

  2. Centro de Energia Nuclear na Agricultura (CENA), USP, Av. Centenário, 303, Piracicaba, SP, 13400-970, Brazil

    Joni Esrom Lima, Vagner Augusto Benedito & Antonio Figueira

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  1. Joni Esrom Lima
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  2. Vagner Augusto Benedito
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Correspondence to Lázaro Eustáquio Pereira Peres.

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Communicated by A. Feher.

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Lima, J.E., Benedito, V.A., Figueira, A. et al. Callus, shoot and hairy root formation in vitro as affected by the sensitivity to auxin and ethylene in tomato mutants. Plant Cell Rep 28, 1169–1177 (2009). https://doi.org/10.1007/s00299-009-0718-y

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  • DOI: https://doi.org/10.1007/s00299-009-0718-y

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