Molecular Genetics and Genomics

, Volume 282, Issue 1, pp 47–64 | Cite as

The peach (Prunus persica [L.] Batsch) homeobox gene KNOPE3, which encodes a class 2 knotted-like transcription factor, is regulated during leaf development and triggered by sugars

  • Giulio Testone
  • Emiliano Condello
  • Ignazio Verde
  • Emilia Caboni
  • Maria Adelaide Iannelli
  • Leonardo Bruno
  • Domenico Mariotti
  • Maria Beatrice Bitonti
  • Donato Giannino
Original Paper


Class 1 KNOTTED1-like transcription factors (KNOX) are known to regulate plant development, whereas information on class 2 KNOX has been limited. The peach KNOPE3 gene was cloned, belonged to a family of few class 2 members and was located at 66 cM in the Prunus spp. G1 linkage-group. The mRNA localization was diversified in leaf, stem, flower and drupe, but recurred in all organ sieves, suggesting a role in sap nutrient transport. During leaf development, the mRNA earliest localized to primordia sieves and subsequently to mesophyll cells of growing leaves. Consistently, its abundance augmented with leaf expansion. The transcription was monitored in leaves responding to darkening, supply and transport block of sugars. It peaked at 4 h after darkness and dropped under prolonged obscurity, showing a similar kinetic to that of sucrose content variation. Feeding leaflets via the transpiration stream caused KNOPE3 up-regulation at 3 h after fructose, glucose and sucrose absorption and at 12 h after sorbitol. In girdling experiments, leaf KNOPE3 was triggered from 6 h onwards along with sucrose and sorbitol raise. Both the phloem-associated expression and sugar-specific gene modulation suggest that KNOPE3 may play a role in sugar translocation during the development of agro-relevant organs such as drupe.


KNOPE3 Transcription factor Peach Gene regulation by light and sugars 



We thank Chiara Nicolodi and Mauro Santini (IBBA-CNR) for technical support, Dr. Samuel Forrest (Clemson University, SC, US) for criticism and English revision. Financial sustains derived from the UNICAL (E.C. was awarded by a PhD scholarship), the project “Eumorfo” of CRA-MIPAF and the CNR Agro-food Department (Director Alcide Bertani). This work is dedicated to the memory of our guide Domenico Mariotti (1948–2008).

Supplementary material

438_2009_445_MOESM1_ESM.doc (36 kb)
Exons are typed in black, introns in italics. Intron consensuses are underlined. Sequences of primers are capitalised; primer names and orientation are shaded in red. Hinc II sites are double-underlined. (DOC 36 kb)
438_2009_445_MOESM2_ESM.ppt (156 kb)
Supplementary material 2 (PPT 156 kb)
438_2009_445_MOESM3_ESM.ppt (121 kb)
Supplementary material 3 (PPT 121 kb)
438_2009_445_MOESM4_ESM.ppt (68 kb)
Supplementary material 4 (PPT 68 kb)
438_2009_445_MOESM5_ESM.ppt (142 kb)
Supplementary material 5 (PPT 142 kb)


  1. An YQ, McDowell JM, Huang S, McKinney EC, Chambliss S, Meagher RB (1996) Strong, constitutive expression of the Arabidopsis ACT2/ACT8 actin subclass in vegetative tissues. Plant J 10:107–121PubMedCrossRefGoogle Scholar
  2. Bassett C (2007) Regulation of gene expression in plants. The role of transcript structure and processing. Springer, HeidelbergGoogle Scholar
  3. Burglin TR (1997) Analysis of TALE superclass homeobox genes (MEIS, PBC, KNOX, Iroquois, TGIF) reveals a novel domain conserved between plants and animals. Nucleic Acids Res 25:4173–4180PubMedCrossRefGoogle Scholar
  4. Cañas L, Busscher M, Angenent G, Beltran J, van Tunen A (1994) Nuclear localization of the petunia MADS box protein FBP1. Plant J 6:597–604CrossRefGoogle Scholar
  5. Chandler J, Wolfgang W (2004) KNAT3 and KNAT4: two KNOX genes control multiple aspects of plant development and are active in the shoot apical meristem. 15th International conference on Arabidopsis research. BerlinGoogle Scholar
  6. Church G, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995PubMedCrossRefGoogle Scholar
  7. Cole M, Nolte C, Werr W (2006) Nuclear import of the transcription factor SHOOT MERISTEMLESS depends on heterodimerization with BLH proteins expressed in discrete sub-domains of the shoot apical meristem of Arabidopsis thaliana. Nucleic Acids Res 34:1281–1292PubMedCrossRefGoogle Scholar
  8. Di Giacomo E, Sestili F, Iannelli MA, Testone G, Mariotti D, Frugis G (2008) Characterization of KNOX genes in Medicago truncatula. Plant Mol Biol 67:135–150PubMedCrossRefGoogle Scholar
  9. Dirlewanger E, Graziano E, Joobeur T, Garriga-Caldere F, Cosson P, Howad W, Arus P (2004) Comparative mapping and marker-assisted selection in Rosaceae fruit crops. Proc Natl Acad Sci USA 101:9891–9896PubMedCrossRefGoogle Scholar
  10. Giannino D, Frugis G, Ticconi C, Florio S, Mele G, Santini L, Cozza R, Bitonti MB, Innocenti A, Mariotti D (2000) Isolation and molecular characterisation of the gene encoding the cytoplasmic ribosomal protein S28 in Prunus persica [L.] Batsch. Mol Gen Genet 263:201–212PubMedCrossRefGoogle Scholar
  11. Giannino D, Condello E, Bruno L, Testone G, Tartarini A, Cozza R, Innocenti AM, Bitonti MB, Mariotti D (2004) The gene geranylgeranyl reductase of peach (Prunus persica [L.] Batsch) is regulated during leaf development and responds differentially to distinct stress factors. J Exp Bot 55:2063–2073PubMedCrossRefGoogle Scholar
  12. Graber JH, Cantor CR, Mohr SC, Smith TF (1999) In silico detection of control signals: mRNA 3′-end-processing sequences in diverse species. Proc Natl Acad Sci USA 96:14055–14060PubMedCrossRefGoogle Scholar
  13. Groover AT, Mansfield SD, DiFazio SP, Dupper G, Fontana JR, Millar R, Wang Y (2006) The Populus homeobox gene ARBORKNOX1 reveals overlapping mechanisms regulating the shoot apical meristem and the vascular cambium. Plant Mol Biol 61:917–932PubMedCrossRefGoogle Scholar
  14. Guo M, Thomas J, Collins G, Timmermans MC (2008) Direct repression of KNOX loci by the ASYMMETRIC LEAVES1 complex of Arabidopsis. Plant Cell 20:48–58PubMedCrossRefGoogle Scholar
  15. Hackbusch J, Richter K, Muller J, Salamini F, Uhrig JF (2005) A central role of Arabidopsis thaliana ovate family proteins in networking and subcellular localization of 3-aa loop extension homeodomain proteins. Proc Natl Acad Sci USA 102:4908–4912PubMedCrossRefGoogle Scholar
  16. Hake S, Smith HM, Holtan H, Magnani E, Mele G, Ramirez J (2004) The role of knox genes in plant development. Annu Rev Cell Dev Biol 20:125–151PubMedCrossRefGoogle Scholar
  17. Harrison J, Moller M, Langdale J, Cronk Q, Hudson A (2005) The role of KNOX genes in the evolution of morphological novelty in Streptocarpus. Plant Cell 17:430–443PubMedCrossRefGoogle Scholar
  18. Hay A, Tsiantis M (2006) The genetic basis for differences in leaf form between Arabidopsis thaliana and its wild relative Cardamine hirsuta. Nat Genet 38:942–947PubMedCrossRefGoogle Scholar
  19. Hay A, Craft J, Tsiantis M (2004) Plant hormones and homeoboxes: bridging the gap? Bioessays 26:395–404PubMedCrossRefGoogle Scholar
  20. Hofer J, Gourlay C, Michael A, Ellis TH (2001) Expression of a class 1 knotted1-like homeobox gene is down-regulated in pea compound leaf primordia. Plant Mol Biol 45:387–398PubMedCrossRefGoogle Scholar
  21. Janssen BJ, Williams A, Chen JJ, Mathern J, Hake S, Sinha N (1998) Isolation and characterization of two knotted-like homeobox genes from tomato. Plant Mol Biol 36:417–425PubMedCrossRefGoogle Scholar
  22. Joobeur T, Viruel M, de Vicente M, Jàuregui B, Ballester J, Dettori M, Verde I, Truco M, Messeguer R, Batlle I, Quarta R, Dirlewanger E, Arùs P (1998) Construction of a saturated linkage map for Prunus using an almond peach F2 progeny. Theor Appl Genet 97:1034–1041CrossRefGoogle Scholar
  23. Kerstetter R, Vollbrecht E, Lowe B, Veit B, Yamaguchi J, Hake S (1994) Sequence analysis and expression patterns divide the maize knotted1-like homeobox genes into two classes. Plant Cell 6:1877–1887PubMedCrossRefGoogle Scholar
  24. Kim JY, Rim Y, Wang J, Jackson D (2005) A novel cell-to-cell trafficking assay indicates that the KNOX homeodomain is necessary and sufficient for intercellular protein and mRNA trafficking. Genes Dev 19:788–793PubMedCrossRefGoogle Scholar
  25. Kimura S, Koenig D, Kang J, Yoong FY, Sinha N (2008) Natural variation in leaf morphology results from mutation of a novel KNOX gene. Curr Biol 18:672–677PubMedCrossRefGoogle Scholar
  26. Kingston RE, Chomczynski P, Sacchi N (2001) Guanidine methods for total RNA preparation. Curr Protoc Mol Biol Chap 4, Unit 4.2Google Scholar
  27. Kitamura-Abe S, Itoh H, Washio T, Tsutsumi A, Tomita M (2004) Characterization of the splice sites in GT-AG and GC-AG introns in higher eukaryotes using full-length cDNAs. J Bioinform Comput Biol 2:309–331PubMedCrossRefGoogle Scholar
  28. Kosambi D (1944) The estimation of map distances from recombination values. Ann Eugen 12:172–175Google Scholar
  29. Krapp A, Quick W, Stitt M (1991) Ribulose-1, 5-bisphosphate carboxylase-oxygenase, other Calvin-cycle enzymes, and chlorophyll decrease when glucose is supplied to mature spinach leaves via the transpiration stream. Planta 186:58–69CrossRefGoogle Scholar
  30. Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln SE, Newberg L (1987) MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174–181PubMedCrossRefGoogle Scholar
  31. Li H, Xu L, Wang H, Yuan Z, Cao X, Yang Z, Zhang D, Xu Y, Huang H (2005) The Putative RNA-dependent RNA polymerase RDR6 acts synergistically with ASYMMETRIC LEAVES1 and 2 to repress BREVIPEDICELLUS and MicroRNA165/166 in Arabidopsis leaf development. Plant Cell 17:2157–2171PubMedCrossRefGoogle Scholar
  32. Lin JF, Wu SH (2004) Molecular events in senescing Arabidopsis leaves. Plant J 39:612–628PubMedCrossRefGoogle Scholar
  33. Lincoln S, Daly M, Lander E (1992) Mapping genes controlling quantitative traits with MAPMAKER/QTL, 2nd edn. Whitehead Institute Technical Report, CambridgeGoogle Scholar
  34. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  35. Lo Bianco R, Rieger M, Sung SJ (1999) Carbohydrate metabolism of vegetative and reproductive sinks in the late-maturing peach cultivar ‘Encore’. Tree Physiol 19:103–109PubMedGoogle Scholar
  36. Magnani E, Hake S (2008) KNOX lost the OX: the Arabidopsis KNATM gene defines a novel class of KNOX transcriptional regulators missing the homeodomain. Plant Cell 20:875–887PubMedCrossRefGoogle Scholar
  37. Maurel K, Leite GB, Bonhomme M, Guilliot A, Rageau R, Petel G, Sakr S (2004) Trophic control of bud break in peach (Prunus persica) trees: a possible role of hexoses. Tree Physiol 24:579–588PubMedGoogle Scholar
  38. Merlo L, Passera C (1991) Changes in carbohydrates and enzyme levels during development of leaves of Prunus persica: a sorbitol synthesizing species. Physiol Plant 83:621–626CrossRefGoogle Scholar
  39. Moing A, Carbonne R, Zipperlin B, Svanella L, Gaudillere J (1997) Phloem loading in peach: sympiastic or apoplastic? Physiol Plant 101:489–496CrossRefGoogle Scholar
  40. Morandi B, Rieger M, Grappadelli LC (2007) Vascular flows and transpiration affect peach (Prunus persica Batsch.) fruit daily growth. J Exp Bot 58:3941–3947PubMedCrossRefGoogle Scholar
  41. Morere-Le Paven MC, Anzala F, Recton A, Limami AM (2007) Differential transcription initiation and alternative RNA splicing of Knox7, a class 2 homeobox gene of maize. Gene 401:71–79PubMedCrossRefGoogle Scholar
  42. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  43. Pagnussat GC, Yu HJ, Sundaresan V (2007) Cell-fate switch of synergid to egg cell in Arabidopsis eostre mutant embryo sacs arises from misexpression of the BEL1-like homeodomain gene BLH1. Plant Cell 19:3578–3592PubMedCrossRefGoogle Scholar
  44. Porra R, Thompson W, Kriedemann P (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophylls standards by atomic absorption spectroscopy. Biochim Biophys Acta 975:384–394CrossRefGoogle Scholar
  45. Price J, Laxmi A, St Martin SK, Jang JC (2004) Global transcription profiling reveals multiple sugar signal transduction mechanisms in Arabidopsis. Plant Cell 16:2128–2150PubMedCrossRefGoogle Scholar
  46. Reiser L, Sanchez-Baracaldo P, Hake S (2000) Knots in the family tree: evolutionary relationships and functions of knox homeobox genes. Plant Mol Biol 42:151–166PubMedCrossRefGoogle Scholar
  47. Rolland F, Baena-Gonzalez E, Sheen J (2006) Sugar sensing and signaling in plants: conserved and novel mechanisms. Annu Rev Plant Biol 57:675–709PubMedCrossRefGoogle Scholar
  48. Sanchez Perez R, Howad W, Dicenta F, Arus P, Martinez-Gomez P (2007) Mapping major genes and quantitative trait loci controlling agronomic traits in almond. Plant Breed 126:310–318CrossRefGoogle Scholar
  49. Schutze K, Harter K, Chaban C (2008) Post-translational regulation of plant bZIP factors. Trends Plant Sci 13:247–255PubMedCrossRefGoogle Scholar
  50. Scofield S, Murray JA (2006) KNOX gene function in plant stem cell niches. Plant Mol Biol 60:929–946PubMedCrossRefGoogle Scholar
  51. Sentoku N, Tamaoki M, Nishimura A, Matsuoka M (1998) The homeobox gene NTH23 of tobacco is expressed in the basal region of leaf primordia. Biochim Biophys Acta 1399:203–208PubMedGoogle Scholar
  52. Serikawa KA, Martinez-Laborda A, Zambryski P (1996) Three knotted1-like homeobox genes in Arabidopsis. Plant Mol Biol 32:673–683PubMedCrossRefGoogle Scholar
  53. Serikawa KA, Martinez-Laborda A, Kim HS, Zambryski PC (1997) Localization of expression of KNAT3, a class 2 knotted1-like gene. Plant J 11:853–861PubMedCrossRefGoogle Scholar
  54. Singer SD, Ashton NW (2007) Revelation of ancestral roles of KNOX genes by a functional analysis of Physcomitrella homologues. Plant Cell Rep 26:2039–2054PubMedCrossRefGoogle Scholar
  55. Smith AM, Zeeman SC, Thorneycroft D, Smith SM (2003) Starch mobilization in leaves. J Exp Bot 54:577–583PubMedCrossRefGoogle Scholar
  56. Soucek P, Klima P, Rekova A, Brzobohaty B (2007) Involvement of hormones and KNOXI genes in early Arabidopsis seedling development. J Exp Bot 58:3797–3810PubMedCrossRefGoogle Scholar
  57. Tamaoki M, Tsugawa H, Minami E, Kayano T, Yamamoto N, Kano-Murakami Y, Matsuoka M (1995) Alternative RNA products from a rice homeobox gene. Plant J 7:927–938PubMedCrossRefGoogle Scholar
  58. Testone G, Bruno L, Condello E, Chiappetta A, Bruno A, Mele G, Tartarini A, Spano L, Innocenti AM, Mariotti D, Bitonti MB, Giannino D (2008) Peach [Prunus persica (L.) Batsch] KNOPE1, a class 1 KNOX orthologue to Arabidopsis BREVIPEDICELLUS/KNAT1, is misexpressed during hyperplasia of leaf curl disease. J Exp Bot 59:389–402PubMedCrossRefGoogle Scholar
  59. Thum KE, Shin MJ, Gutierrez RA, Mukherjee I, Katari MS, Nero D, Shasha D, Coruzzi GM (2008) An integrated genetic, genomic and systems approach defines gene networks regulated by the interaction of light and carbon signaling pathways in Arabidopsis. BMC Syst Biol 2:31PubMedCrossRefGoogle Scholar
  60. Truernit E, Siemering KR, Hodge S, Grbic V, Haseloff J (2006) A map of KNAT gene expression in the Arabidopsis root. Plant Mol Biol 60:1–20PubMedCrossRefGoogle Scholar
  61. Vemmos S, Goldwin G (1994) The photosynthetic activity of Cox’s Orange Pippin apple flowers in relation to fruit setting. Ann Botany 73:385–391CrossRefGoogle Scholar
  62. Watillon B, Kettmann R, Boxus P, Burny A (1993) Developmental and circadian pattern of rubisco activase mRNA accumulation in apple plants. Plant Mol Biol 23:501–509PubMedCrossRefGoogle Scholar
  63. Watillon B, Kettmann R, Boxus P, Burny A (1997) Knotted1-like homeobox genes are expressed during apple tree (Malus domestica [L.] Borkh) growth and development. Plant Mol Biol 33:757–763PubMedCrossRefGoogle Scholar
  64. Weaver LM, Amasino RM (2001) Senescence is induced in individually darkened Arabidopsis leaves, but inhibited in whole darkened plants. Plant Physiol 127:876–886PubMedCrossRefGoogle Scholar
  65. Yamaki S (1980) Properties and function of sorbitol-6-phosphate dehydrogenase, sorbitol dehydrogenase and sorbitol oxidase in fruit and cotyledon of apple Malus pumila Mill. var.domestica Schneid. J Japan Soc Hort Sci 49:429–434CrossRefGoogle Scholar
  66. Yoshii A, Shimizu T, Yoshida A, Hamada K, Sakurai K, Yamaji Y, Suzuki M, Namba S, Hibi T (2008) NTH201, a novel class II KNOTTED1-like protein, facilitates the cell-to-cell movement of Tobacco mosaic virus in tobacco. Mol Plant Microbe Interact 21:586–596PubMedCrossRefGoogle Scholar
  67. Zanchin A, Bonghi C, Casadoro G, Ramina A, Rascio N (1994) Cell enlargement and cell separation during peach fruit development. Int J Plant Sci 155:49–56CrossRefGoogle Scholar
  68. Zhong R, Richardson EA, Ye ZH (2007) The MYB46 transcription factor is a direct target of SND1 and regulates secondary wall biosynthesis in Arabidopsis. Plant Cell 19:2776–2792PubMedCrossRefGoogle Scholar
  69. Zhou R, Cheng L, Dandekar AM (2006) Down-regulation of sorbitol dehydrogenase and up-regulation of sucrose synthase in shoot tips of the transgenic apple trees with decreased sorbitol synthesis. J Exp Bot 57:3647–3657PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Giulio Testone
    • 1
  • Emiliano Condello
    • 1
  • Ignazio Verde
    • 2
  • Emilia Caboni
    • 2
  • Maria Adelaide Iannelli
    • 1
  • Leonardo Bruno
    • 3
  • Domenico Mariotti
    • 1
  • Maria Beatrice Bitonti
    • 3
  • Donato Giannino
    • 1
  1. 1.Institute of Agricultural Biology and BiotechnologyNational Research Council of Italy (CNR)Monterotondo Scalo, RomeItaly
  2. 2.Fruit Tree Research CenterAgriculture Research Council (CRA)RomeItaly
  3. 3.Department of EcologyUniversity of CalabriaArcavacata di Rende, CosenzaItaly

Personalised recommendations