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Tree Physiology Advance Access originally published online on January 30, 2009
Tree Physiology 2009 29(4):569-577; doi:10.1093/treephys/tpn049
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© The Author 2009. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Characterization of gibberellin 20-oxidases in the citrus hybrid Carrizo citrange

Laura Huerta1, Andres Garcia-Lor1 and Jose L. Garcia-Martinez1,2

1 Instituto de Biología Molecular y Celular de Plantas, Universidad Politécnica de Valencia-CSIC, Avda de los Naranjos s/n, 46022 Valencia, Spain
2 Corresponding author (jlgarcim{at}ibmcp.upv.es)


  Abstract

CcGA20ox1 is a gene encoding a GA 20-oxidase, a gibberellin (GA) biosynthetic enzyme, previously isolated from the citrus hybrid Carrizo citrange (Citrus sinensis (L.) Osbeck x Poncirus trifoliata (L.) Raf.). Southern blot analysis of genomic DNA of Carrizo citrange with CcGA20ox1 suggested the presence in the hybrid of another gene encoding another GA 20-oxidase. A cDNA clone from this new gene (CcGA20ox2) was isolated using RNA from the other parent C. sinensis. CcGA20ox2 encoded a protein of 372 amino acids that showed 67.1% identity with CcGA20ox1, and its expression product catalyzed the in vitro conversion of GA12 to GA9, confirming that it corresponds to another active GA20ox. Amplification of genomic DNA and isolation of genomic clones of CcGA20ox1 and CcGA20ox2 revealed that the parental sources of these genes in the hybrid were P. trifoliata and C. sinensis, respectively. The sequences of CcGA20ox1 and CcGA20ox2 showed that both genes contained two introns, which are also conserved in GA20ox genes of other species like Arabidopsis thaliana L., Pisum sativum L. and Solanum lycopersicum L. Determination of transcript levels in the Carrizo citrange hybrid by quantitative real-time polymerase chain reaction showed that CcGA20ox1 was expressed mainly in internodes, leaves and seeds, and CcGA20ox2 in flower buds and flowers at anthesis, with the genes having similar transcript levels in young developing fruits.

Keywords: citrus rootstock, Citrus sinensis, GA 20-oxidase, gibberellins, Poncirus trifoliata

Received July 21, 2008; Accepted December 2, 2008


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