Planta

, Volume 226, Issue 4, pp 877–888 | Cite as

Tomato fruit set driven by pollination or by the parthenocarpic fruit allele are mediated by transcriptionally regulated gibberellin biosynthesis

  • Irene Olimpieri
  • Francesca Siligato
  • Riccardo Caccia
  • Gian Piero Soressi
  • Andrea Mazzucato
  • Lorenzo Mariotti
  • Nello Ceccarelli
Original Article

Abstract

We investigated the role of gibberellins (GAs) in the phenotype of parthenocarpic fruit (pat), a recessive mutation conferring parthenocarpy in tomato (Solanum lycopersicum L.). Novel phenotypes that parallel those reported in plants repeatedly treated with gibberellic acid or having a GA-constitutive response indicate that the pat mutant probably expresses high levels of GA. The retained sensitivity to the GA-biosynthesis inhibitor paclobutrazol reveals that this condition is dependent on GA biosynthesis. Expression analysis of genes encoding key enzymes involved in GA biosynthesis shows that in normal tomato ovaries, the GA20ox1 transcript is in low copy number before anthesis and only pollination and fertilization increase its transcription levels and, thus, GA biosynthesis. In the unpollinated ovaries of the pat mutant, this mechanism is de-regulated and GA20ox1 is constitutively expressed, indicating that a high GA concentration could play a part in the parthenocarpic phenotype. The levels of endogenous GAs measured in the floral organs of the pat mutant support such a hypothesis. Collectively, the data indicate that transcriptional regulation of GA20ox1 mediates pollination-induced fruit set in tomato and that parthenocarpy in pat results from the mis-regulation of this mechanism. As genes involved in the control of GA synthesis (LeT6, LeT12 and LeCUC2) and response (SPY) are also altered in the pat ovary, it is suggested that the pat mutation affects a regulatory gene located upstream of the control of fruit set exerted by GAs.

Keywords

Fruit set GA 20-oxidase Parthenocarpy Tomato 

Abbreviations

CUC2

Cup-shaped cotyledon2

DAP

Days after pollination

GA

Gibberellin

GA3

Gibberellic acid

GA20ox

GA 20-oxidase

GA3ox

GA 3-oxidase

GC-MS/MS

Chromatography–tandem mass spectrometry

HAE

Hours after emasculation

HAP

Hours after pollination

HBP

Hours before pollination

HP

Hand-pollinated

IAA

Indole-3-acetic acid

KNOX

Knotted-like homeobox genes

OP

Open-pollinated

PAC

Paclobutrazol

Pat

Parthenocarpic fruit

RT

Reverse transcription

SPY

Spindly

WT

Wild type

Supplementary material

425_2007_533_MOESM1_ESM.ppt (39 kb)
Fig. S1 The principal pathways of gibberellin (GA) metabolism in higher plants, including the non-early-13-hydroxylated pathway (left branch) and the early-13-hydroxylated pathway (right branch). Arrows connect the precursor with the product of each step and circled numbers refers to the enzymes catalysing the reactions as follows: (1) ent-copalyl diphosphate synthase; (2) ent-kaurene synthase; (3) ent-kaurene 19-oxidase; (4) kaurenoic acid oxidase; (5) GA 13-hydroxylase; (6) GA 20-oxidase (GA20ox); (7) GA 3β-hydroxylase (GA3ox); (8) GA 2-oxidase (GA2ox). Those components of the early-13-hydroxylated pathway that have been monitored in this study are highlighted. (Modified from Hedden and Phillips, 2000). GGPP, geranylgeranyl pyrophosphate (DOC 39 kb)
425_2007_533_MOESM2_ESM.ppt (37 kb)
Fig. S2 Expression analysis of genes involved in GA synthesis and response in WT (white bars) and pat mutant (black bars) tomato ovaries at the time of flower opening (stage 3). RNA was isolated and reverse transcribed using oligo(dT) and Moloney murine leukemia virus-reverse transcriptase. The cDNAs generated were subsequently used in a 25 ml PCR reaction in the presence of primers specific for the studied genes or the ACTIN control. The RT-PCR products were separated on 1.5% (w/v) agarose gels stained with ethidium bromide. Expression data are reported as estimates of relative mRNA amount derived from the ratio between the quantitative values of PCR band intensity for the target gene and the actin control measured by the Gel Analyzer procedure of the ImageJ software (http://rsb.info.nih.gov/ij/). Data points are means of three biological replicates ±SE and * indicates significant differences for P£0.05 between the WT and pat mutant ovaries after one-way analysis of variance (DOC 37 kb)
425_2007_533_MOESM3_ESM.doc (32 kb)
Table S1. Sets of RT-PCR primers and PCR conditions used to amplify tomato gene-specific regions of coding sequences involved in GA biosynthesis and response (DOC 33 kb)

References

  1. Abdul KS, Harris GP (1978) Control of flower number in the first inflorescenze of tomato (Lycopersicon esculentum Mill.): the role of gibberellins. Ann Bot 42:1361–1367Google Scholar
  2. Aida M, Ishida T, Tasaka M (1999) Shoot apical meristem and cotyledon formation during Arabidopsis embryogenesis: interaction among the CUP-SHAPED COTYLEDON and SHOOT MERISTEMLESS genes. Development 126:1563–1570PubMedGoogle Scholar
  3. Angenent GC, Stuurman J, Snowden KC, Koes R (2005) Use of Petunia to unravel plant meristem functioning. Trends Plant Sci 10:243–250PubMedCrossRefGoogle Scholar
  4. Ben-Cheikh W, Perez-Botella J, Tadeo FR, Talon M, Primo-Millo E (1997) Pollination increases gibberellin levels in developing ovaries of seeded varieties of Citrus. Plant Physiol 114:557–564PubMedGoogle Scholar
  5. Beraldi D, Picarella ME, Soressi GP, Mazzucato A (2004) Fine mapping of the parthenocarpic fruit (pat) mutation in tomato. Theor Appl Genet 108:209–216PubMedCrossRefGoogle Scholar
  6. Fos M, Nuez F, García-Martínez J (2000) The gene pat-2, which induces natural parhenocarpy, alters the gibberellin content in unpollinated tomato ovaries. Plant Physiol 122:471–479PubMedCrossRefGoogle Scholar
  7. Fos M, Proaño K, Nuez F, García-Martínez JL (2001) Role of gibberellins in parthenocarpic fruit development induced by the genetic system pat-3/pat-4 in tomato. Physiol Plant 111:545–550PubMedCrossRefGoogle Scholar
  8. García-Martínez JL, López-Diaz I, Sánchez-Beltrán MJ, Phillips AL, Ward DA, Gaskin P, Hedden P (1997) Isolation and transcript analysis of gibberellin 20-oxidase genes in pea and bean in relation to fruit development. Plant Mol Biol 33:1073–1084PubMedCrossRefGoogle Scholar
  9. Gillaspy G, Ben-David H, Gruissem W (1993) Fruits: a developmental perspective. Plant Cell 5:1439–1451PubMedCrossRefGoogle Scholar
  10. Goetz M, Vivian-Smith A, Johnson SD, Koltunow AM (2006) AUXIN RESPONSE FACTOR8 is a negative regulator of fruit initiation in Arabidopsis. Plant Cell 18:1873–1886PubMedCrossRefGoogle Scholar
  11. Gorguet B, van Heusden AW, Lindhout P (2005) Parthenocarpic fruit development in tomato. Plant Biol 7:131–139PubMedCrossRefGoogle Scholar
  12. Greb T, Schmitz G, Theres K (2002) Isolation and characterization of the Spindly homologue from tomato. J Exp Bot 53:1829–1830PubMedCrossRefGoogle Scholar
  13. Harrison AL (1955) New “mouse-eared” mutant from var Rutgers. Rep Tomato Genet Coop 5:18Google Scholar
  14. Hay A, Kaur H, Phillips A, Hedden P, Hake S, Tsiantis M (2002) The gibberellin pathway mediates KNOTTED1-type homeobox function in plants with different body plans. Curr Biol 12:1557–1565PubMedCrossRefGoogle Scholar
  15. Hisamatsu T, King RW, Helliwell CA, Koshioka M (2005) The involvement of gibberellin 20-oxidase genes in phytochrome-regulated petiole elongation of Arabidopsis. Plant Physiol 138:1106–1116PubMedCrossRefGoogle Scholar
  16. Ishida T, Aida M, Takada S, Tasaka M (2000) Involvement of CUP-SHAPED COTYLEDON genes in gynoecium and ovule development in Arabidopsis thaliana. Plant Cell Physiol 41:60–67PubMedGoogle Scholar
  17. Jacobsen SE, Olszewski NE (1993) Mutations at the SPINDLY locus of Arabidopsis alter gibberellin signal transduction. Plant Cell 5:887–896PubMedCrossRefGoogle Scholar
  18. Jacobsen SE, Binkowski KA, Olszewski NE (1996) SPINDLY, a tetratricopeptide repeat protein involved in gibberellin signal transduction in Arabidopsis. Proc Natl Acad Sci USA 93:9292–9296PubMedCrossRefGoogle Scholar
  19. Janssen B-J, Williams A, Chen J-J, Mathern J, Hake S, Sinha N (1998a) Isolation and characterization of two knotted-like homeobox genes from tomato. Plant Mol Biol 36:417–425PubMedCrossRefGoogle Scholar
  20. Janssen B-J, Lund L, Sinha N (1998b) Overexpression of a homeobox gene, LeT6, reveals indeterminate features in the tomato compound leaf. Plant Physiol 117:771–786PubMedCrossRefGoogle Scholar
  21. Jasinski S, Piazza P, Craft J, Hay A, Wooley L, Rieu I, Phillips A, Hedden P, Tsiantis M (2005) KNOX action in Arabidopsis is mediated by coordinate regulation of cytokinin and gibberellin activities. Curr Biol 15:1560–1565PubMedCrossRefGoogle Scholar
  22. Kamiya Y, García-Martínez JL (1999) Regulation of gibberellin biosynthesis by light. Curr Opin Plant Biol 2:398–403PubMedCrossRefGoogle Scholar
  23. Kang H-G, Jun S-H, Kim J, Kawaide H, Kamiya Y, An G (1999) Cloning and molecular analyses of a gibberellin 20-oxidase gene expressed specifically in developing seeds of watermelon. Plant Physiol 121:373–382PubMedCrossRefGoogle Scholar
  24. Kataoka K, Uemachi A, Yazawa S (2003) Fruit growth and pseudoembryo development affected by uniconazole, an inhibitor of gibberellin biosynthesis, in pat-2 and auxin-induced parthenocarpic tomato fruits. Sci Hortic 98:9–16CrossRefGoogle Scholar
  25. Kataoka K, Okita H, Uemachi A, Yazawa S (2004) A pseudoembryo highly stainable with toluidine blue may induce fruit growth of parthenocarpic tomato. Acta Hortic 637:213–221Google Scholar
  26. Kerstetter RA, Laudencia-Chingcuanco D, Smith LG, Hake S (1997) Loss-of-function mutations in the maize homeobox gene, knotted1, are defective in shoot meristem maintenance. Development 124:3045–3054PubMedGoogle Scholar
  27. King PJ (1988) Plant hormone mutants. Trends Genet 4:157–162PubMedCrossRefGoogle Scholar
  28. Koltunow AM, Vivian-Smith A, Tucker MR, Paech N (2002) The central role of the ovule in apomixis and parthenocarpy. In: O’Neill SD, Roberts JA (eds) Plant reproduction. Academic, Sheffield, pp 221–256Google Scholar
  29. Koshioka M, Nishijima T, Yamazaki H, Nonaka M, Mander LN (1994) Analysis of gibberellins in growing fruits of Lycopersicon esculentum after pollination or treatment with 4-chlorophenoxyacetic acid. J Hortic Sci 69:171–179Google Scholar
  30. Mapelli S, Frova C, Torti G, Soressi GP (1978) Relationship between set, development and activities of growt regulators in tomato fruits. Plant Cell Physiol 19:1281–1288Google Scholar
  31. Mazzucato A, Taddei AR, Soressi GP (1998) The parthenocarpic fruit (pat) mutant of tomato (Lycopersicon esculentum Mill.) sets seedless fruits and has aberrant anther and ovule development. Development 125:107–114PubMedGoogle Scholar
  32. Mazzucato A, Testa G, Biancari T, Soressi GP (1999) Effect of gibberellic acid treatments, environmental conditions, and genetic background on the expression of the parthenocarpic fruit mutation in tomato. Protoplasma 208:18–25CrossRefGoogle Scholar
  33. Nitsch J (1970) Hormonal factors in growth and development. In: AC Hulme (ed) The biochemistry of fruits and their products, vol II. Academic, London, pp 427–472Google Scholar
  34. O’Neill SD (1997) Pollination regulation of flower development. Annu Rev Plant Physiol Plant Mol Biol 48:547–574PubMedCrossRefGoogle Scholar
  35. Oikawa T, Koshioka M, Kojima K, Yoshida H, Kawata M (2004) A role of OsGA20ox1, encoding an isoform of gibberellin 20-oxidase, for regulation of plant stature in rice. Plant Mol Biol 55:687–700PubMedCrossRefGoogle Scholar
  36. Olszewski N, Sun TP, Gubler F (2002) Gibberellin signaling: biosynthesis, catabolism, and response pathways. Plant Cell 14(Suppl):S61–S80PubMedGoogle Scholar
  37. Ozga JA, Reinecke DM (2003) Hormonal interactions in fruit development. J Plant Growth Regul 22:73–81CrossRefGoogle Scholar
  38. Ozga JA, Yu J, Reinecke DM (2003) Pollination-, development-, and auxin-specific regulation of gibberellin 3β-hydroxilase gene expression in pea fruit and seeds. Plant Physiol 131:1137–1146PubMedCrossRefGoogle Scholar
  39. Parnis A, Cohen O, Gutfinger T, Hareven D, Zamir D, Lifschitz E (1997) The dominant developmental mutants of tomato, Mouse-ear and Curl, are associated with distinct modes of abnormal transcriptional regulation of a Knotted gene. Plant Cell 9:2143–2158PubMedCrossRefGoogle Scholar
  40. Picciarelli P, Piaggesi L, Ceccarelli N, Guglielminetti L, Alpi A (1994) Gibberellins in suspensor, embryo and integuments from very young seeds of Phaseolus coccineus L. Plant Growth Regul 14:183–185CrossRefGoogle Scholar
  41. Ray A, Robinson-Beers K, Ray S, Baker SC, Lang JD, Preuss D, Milligan SB, Gasser CS (1994) Arabidopsis floral homeotic gene BELL (BEL1) controls ovule development through negative regulation of AGAMOUS gene (AG). Proc Natl Acad Sci USA 91:5761–5765PubMedCrossRefGoogle Scholar
  42. Rebers M, Kaneta T, Kawaide H, Yamaguchi S, Yang YY, Imai R, Sekimoto H, Kamiya Y (1999) Regulation of gibberellin biosynthesis genes during flower and early fruit development of tomato. Plant J 17:241–250PubMedCrossRefGoogle Scholar
  43. Ross JJ, O’Neill DP, Smith JJ, Kerckhoffs LHJ, Elliot RC (2000) Evidence that auxin promotes gibberellin A1 biosynthesis in pea. Plant J 21:547–552PubMedCrossRefGoogle Scholar
  44. Sawhney VK, Greyson RI (1971) Induction of multilocular ovary in tomato by gibberellic acid. J Am Soc Hortic Sci 96:196–198Google Scholar
  45. Schwabe WW, Mills JJ (1981) Hormones and parthenocarpic fruit set. Hort Abstr 51:661–698Google Scholar
  46. Schumacher K, Schmitt T, Rossberg M, Schmitz G, Theres K (1999) The lateral suppressor (Ls) gene of tomato encodes a new member of the VHIID protein family. Proc Natl Acad Sci USA 96:290–295PubMedCrossRefGoogle Scholar
  47. Solfanelli C, Ceron F, Paolicchi F, Giorgetti L, Geri C, Ceccarelli N, Kamiya Y, Picciarelli P (2005) Expression of two genes encoding gibberellin 2- and 3-oxidases in developing seeds of Phaseolus coccineus. Plant Cell Physiol 46:1116–1124PubMedCrossRefGoogle Scholar
  48. Sun TP, Gubler F (2004) Molecular mechanism of gibberellin signaling in plants. Annu Rev Plant Biol 55:197–223PubMedCrossRefGoogle Scholar
  49. Talon M, Koornneef M, Zeevaart JAD (1990) Endogenous gibberellins in Arabidopsis thaliana and possible steps blocked in the biosynthetic pathways of the semidwarf ga4 and ga5 mutants. Proc Natl Acad Sci USA 87:7983–7987PubMedCrossRefGoogle Scholar
  50. Talon M, Zacarias L, Primo-Millo E (1992) Gibberellins and parthenocarpic ability in developing ovaries of seedless mandarins. Plant Physiol 99:1575–1581PubMedCrossRefGoogle Scholar
  51. Tanaka-Ueguchi M, Itoh H, Oyama N, Koshioka M, Matsuoka M (1998) Over-expression of a tobacco homeobox gene, NTH15, decreases the expression of a gibberellin biosynthetic gene encoding GA 20-oxidase. Plant J 15:391–400PubMedCrossRefGoogle Scholar
  52. Testa G, Caccia R, Tilesi F, Soressi GP, Mazzucato A (2002) Sequencing and characterization of tomato genes putatively involved in fruit set and early development. Sex Plant Reprod 14:269–277CrossRefGoogle Scholar
  53. Varoquaux F, Blanvillain R, Delseny M, Gallois P (2000) Less is better: new approaches for seedless fruit production. Trends Biotechnol 18:233–242PubMedCrossRefGoogle Scholar
  54. Vivian-Smith A, Luo M, Chaudhury A, Koltunow A (2001) Fruit development is actively restricted in the absence of fertilization in Arabidopsis. Development 128:2321–2331PubMedGoogle Scholar
  55. Wang H, Jones B, Li Z, Frasse P, Delalande C, Regad F, Chaabouni S, Latché A, Pech J-C, Bouzayen M (2005) The tomato Aux/IAA transcription factor IAA9 is involved in fruit development and leaf morphogenesis. Plant Cell 17:2676–2692PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Irene Olimpieri
    • 1
  • Francesca Siligato
    • 1
  • Riccardo Caccia
    • 1
  • Gian Piero Soressi
    • 1
  • Andrea Mazzucato
    • 1
  • Lorenzo Mariotti
    • 2
  • Nello Ceccarelli
    • 2
  1. 1.Dipartimento di Agrobiologia e Agrochimica, Sezione di GeneticaUniversità degli Studi della TusciaViterboItaly
  2. 2.Dipartimento di Biologia delle Piante AgrarieUniversità degli Studi di PisaPisaItaly

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