title: Seed-Specific Stable Expression of the α-AI1 Inhibitor in Coffee Grains and theIn VivoImplications for the Development of the Coffee Berry Borer.
abstract: Genetic transformation of coffee (Coffeaspp.), the second most traded commodity worldwide, is an alternative approach to introducing features that cannot be introgressed by traditional crossings. The transgenic stability, heritability and quantitative and spatial expression patterns of the seed-specific promoter phytohemagglutinin (PHA-L) fromPhaseolus vulgariswere characterized in genetically modifiedC. arabicaexpressing the α-amylase inhibitor-1 (α-AI1) gene. The α-AI1 inhibitor shows considerable activity toward digestive enzymes of the coffee berry borer (CBB)Hypothenemus hampei. This insect pest expends its life cycle almost entirely in coffee berries. Transgene containment in the fruit is important to meeting food and environmental safety requirements for releasing genetically modified (GM) crops. PCR analysis of T2 coffee plants showed a Mendelian single-copy segregation pattern. Ectopic transgene expression was only detected in coffee grains, as demonstrated by reverse transcription-PCR analysis of different plant tissues. An intense immunocytochemical signal associated with α-AI1 protein expression was localized to endospermic cells. In addition, a delay in the larval development of CBB was observed after challenging transgenic coffee seeds with the insect. These results indicate that the PHA-L promoter might be a useful tool in coffee for the seed-specific expression of genes related to coffee bean productivity, quality and pest protection. The biotechnological applicability of theα-AI1gene for controlling CBB is also discussed. This work is the first report showing a seed-specific transgene expression in coffee plants.
authors.affiliation: Embrapa Recursos Genéticos e Biotecnologia, 70770-917 Brasília, DF Brazil.
authors.forename: Érika V S
authors.initials: ÉV
authors.lastname: Albuquerque
authors.affiliation: Embrapa Recursos Genéticos e Biotecnologia, 70770-917 Brasília, DF Brazil ; Universidade Católica de Brasília, 70790-160 Brasília, DF Brazil.
authors.forename: Caroline A
authors.initials: CA
authors.lastname: Bezerra
authors.affiliation: Universidad del Atlántico, km 7, Barranquilla, Colombia.
authors.forename: Juan V
authors.initials: JV
authors.lastname: Romero
authors.affiliation: Universidad del Atlántico, km 7, Barranquilla, Colombia.
authors.forename: Jorge W A
authors.initials: JW
authors.lastname: Valencia
authors.affiliation: Facultad de Ciencias Agropecuarias, Universidad de Caldas, Apartado aéreo 275, Manizales, Colombia.
authors.forename: Arnubio
authors.initials: A
authors.lastname: Valencia-Jiménez
authors.affiliation: Embrapa Recursos Genéticos e Biotecnologia, 70770-917 Brasília, DF Brazil ; Universidade Católica de Brasília, 70790-160 Brasília, DF Brazil.
authors.forename: Lucas M
authors.initials: LM
authors.lastname: Pimenta
authors.affiliation: Universidade Católica de Brasília, 70790-160 Brasília, DF Brazil.
authors.forename: Aulus E A D
authors.initials: AE
authors.lastname: Barbosa
authors.affiliation: Embrapa Recursos Genéticos e Biotecnologia, 70770-917 Brasília, DF Brazil.
authors.forename: Maria C M
authors.initials: MC
authors.lastname: Silva
authors.affiliation: IAPAR, Instituto Agronômico do Paraná, 86047-902 Londrina, Brazil.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Meneguim
authors.affiliation: EPAMIG, Empresa de Pesquisa Agropecuária de Minas Gerais, 38001-970 Uberaba, MG Brazil.
authors.forename: Maria Eugênia L
authors.initials: ME
authors.lastname: Sá
authors.affiliation: INRA, Institut National de la Recherche Agronomique, Plant, Health and Environment, Plant-Nematodes Interaction Team, UMR 1355 ISA/Centre National de la Recherche Scientifique, Sophia-Antipolis, France.
authors.forename: Gilbert
authors.initials: G
authors.lastname: Engler
authors.affiliation: INRA, Institut National de la Recherche Agronomique, Plant, Health and Environment, Plant-Nematodes Interaction Team, UMR 1355 ISA/Centre National de la Recherche Scientifique, Sophia-Antipolis, France.
authors.forename: Janice
authors.initials: J
authors.lastname: de Almeida-Engler
authors.affiliation: IRD, Institut de Recherche pour le Développement UMR 186 - IRD/CIRAD/UM2 Résistance des Plantes aux Bio-agresseurs, 34394 Montpellier Cedex 5, France.
authors.forename: Diana
authors.initials: D
authors.lastname: Fernandez
authors.affiliation: Embrapa Recursos Genéticos e Biotecnologia, 70770-917 Brasília, DF Brazil ; Universidade Católica de Brasília, 70790-160 Brasília, DF Brazil.
authors.forename: Maria F
authors.initials: MF
authors.lastname: Grossi-de-Sá
keywords: Coffea arabica; Fruit-specific expression; Genetically modified plants; Hypothenemus hampei; Immunolocalization