Genetic transformation of sweet oranges with the <i>D4E1</i> gene  driven by the AtPP2 promoter

Lísia Borges Attílio; Francisco de Assis Alves Mourão Filho; Ricardo Harakava; Tatiane Loureiro da Silva; Luzia Yuriko Miyata; Liliane Cristina Libório Stipp; Beatriz Madalena Januzzi Mendes

doi:10.1590/S1678-3921.pab2013.v48.14840

Authors

Lísia Borges Attílio Universidade de São Paulo Escola Superior de Agricultura "Luiz de Queiroz"
Francisco de Assis Alves Mourão Filho Universidade de São Paulo Escola Superior de Agricultura "Luiz de Queiroz"
Ricardo Harakava Institulo Biológico
Tatiane Loureiro da Silva Universidade de São Paulo Escola Superior de Agricultura "Luiz de Queiroz"
Luzia Yuriko Miyata Universidade de São Paulo Escola Superior de Agricultura "Luiz de Queiroz"
Liliane Cristina Libório Stipp Universidade de São Paulo Escola Superior de Agricultura "Luiz de Queiroz"
Beatriz Madalena Januzzi Mendes Universidade de São Paulo Centro de Energia Nuclear na Agricultura

DOI:

https://doi.org/10.1590/S1678-3921.pab2013.v48.14840

Keywords:

Citrus sinensis, antimicrobial peptide, tissue‑specific promoter

Abstract

The objective of this work was to produce transgenic 'Pêra' and 'Valência' sweet orange plants using the D4E1 gene driven by the Arabidopsis thaliana phloem protein (AtPP2) promoter and to quantify transgene expression in different transformation events. Genetic transformation experiments were carried out with epicotyl segments co‑cultivated with Agrobacterium tumefaciens. Six plants from 'Pêra' sweet orange and seven plants from 'Valência' sweet orange were confirmed as different transgenic events by means of the polymerase chain reaction (PCR) and the Southern blot techniques. Transgene expression was quantified using real‑time quantitative PCR. D4E1 gene expression levels vary from 5 up to 50 times among different transformation events.