Identification of Differentially Expressed Genes During Embryogenesis in Maritime Pine (Pinus pinaster)

 

Sónia Gonçalves*¹, John Cairney**, Margarida Oliveira*** and Célia Miguel****

*PhD Student

Instituto de Tecnologia Química e Biológica. Quinta do Marquês, 2784-505 Oeiras

**Associate Professor

School of Biology and Institute of Paper Science and Technology, 500 10^th Street Atlanta, NW 30332-0620, USA

***Assistant Professor

Faculdade de Ciências de Lisboa. Departamento Biologia Vegetal. Campo Grande 1749-016 Lisboa, Portugal/Instituto de Tecnologia Química e Biológica, Quinta do Marquês, 2784-505 Oeiras

****Post-Doctoral Researcher

Instituto de Biologia Experimental e Tecnológica. Quinta do Marquês, 2784-505 Oeiras

 

 

Abstract. Differential display of Poly (A⁺) RNA isolated from zygotic embryos of maritime pine (Pinus pinaster) in 9 stages of development was used to isolate several cDNAs of differentially expressed genes during embryo development. Many cDNA sequences share low similarities to any known DNA or protein sequences but 5 transcripts are predicted to encode homologues of DNA-binding proteins, GTP-binding proteins, lipid transferases and dehydrogenases. A full-length cDNA clone was isolated for a gene (named PpAAI-LTSS 1) whose expression appears to be specific to middle stages of development. The full-length cDNA was also isolated for two clones (named PpRab1 and PpCR1) specific to an early stage of embryo development.

Key words: pine; embryo development; gene expression

 

Identificação dos Genes Diferencialmente Expressos Durante a Embriogénese no Pinho Marítimo (Pinus pinaster)

Sumário. A técnica de "differential display" foi utilizada em RNA mensageiro extraído de embriões zigóticos de pinheiro-bravo em 9 estádios de desenvolvimento, de modo a isolar fragmentos de cDNA correspondentes a genes diferencialmente expressos durante o desenvolvimento do embrião. Muitos destes fragmentos possuem baixas homologias com sequências de DNA ou proteicas conhecidas, mas cinco transcritos codificam para homólogos de proteínas de ligação ao DNA, proteínas de ligação ao GTP, transferases lipídicas e desidrogenases. Um clone de cDNA de sequência completa foi isolado para um gene (denominado PpAAI-LTSS1), cuja expressão parece ser específica dos estádios intermédios de desenvolvimento. A sequência completa de cDNA foi também obtida para outros dois clones (denominados PpRab1 e PpCR1), específicos dos estádios iniciais de desenvolvimento do embrião.

Palavras-chave: pinheiro; desenvolvimento embrionário; expressão génica

 

Identification des Gènes Différentiellement Exprimés Pendant l'Embryogenèse en Pin Maritime (Pinus pinaster)

Résumé. Le criblage différentiel d'RNA messagers isolés d'embryons zygotiques de pin maritime (Pinus pinaster) à 9 stades différents du développement a été utilisé pour identifier des gènes dont l'expression varie au cours de l'embryogenèse. Une partie des cDNA isolés présentent une faible homologie avec des séquences connues. Cinq transcrits sont homologues à des gènes codant pour des protéines de liaison à l'DNA et au GTP, à des protéines de transfert de lipides et à des déshydrogénases. Un cDNA complet (nommé PpAAI-LTSS 1) dont l'expression semble être spécifique de la phase intermédiaire du développement a été cloné ainsi que deux autres cDNA complets (PpRab1 et PpCR1) exprimés spécifiquement dans les phases précoces du développement.

Mots clés: pin; développement de l'embryon; expression des gènes

 

 

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Entregue para publicação em Abril de 2004

Aceite para publicação em Março de 2005

 

¹ 1º Autor E-mail: sonia@itqb.unl.pt