RefDoc
Haut

Faire une nouvelle recherche
Make a new search
Lancer la recherche


Titre du document / Document title

Agrobacterium-mediated transformation of Pinus radiata organogenic tissue using vacuum-infiltration

Auteur(s) / Author(s)

CHARITY J. A. (1) ; HOLLAND L. (1) ; DONALDSON S. S. (1) ; GRACE L. (1) ; WALTER C. (1) ;

Affiliation(s) du ou des auteurs / Author(s) Affiliation(s)

(1) New Zealand Forest Research Institute Ltd, Private Bag 3020, Rotorua, NOUVELLE-ZELANDE

Résumé / Abstract

An Agrobacterium tumefaciens-mediated transformation protocol was developed for detached cotyledons of Pinus radiata zygotic embryos resulting in up to 55% of cotyledons transiently expressing the reporter gene uidA. Transient expression of uidA was improved when detached cotyledons were pre-cultured on half strength medium containing cytokinin for 7 days, wounded by vortexing and then vacuum-infiltrated in a solution of A. tumefaciens. The transformation protocol was applied both to cotyledons and also to the apical meristematic dome which was the portion of the embryo remaining after cotyledons were detached, and from which the apical shoot and axillary shoots regenerate. Molecular analysis of putatively transformed shoots regenerated either adventitiously from cotyledons or via axillary shoots from apical domes, indicated the presence of uidA and nptll genes by PCR in some of these shoots. Biochemical analysis of putatively transformed shoots using nptII ELISA indicated that they contained the nptII enzyme. However, Southern hybridisation indicated stable integration of nptII only in one shoot which was regenerated from an apical dome. Shoots regenerated from cotyledons appeared to exhibit chimeric expression and were not stably transformed. Based on a comparison of time for regeneration, technical difficulty, molecular and biochemical analysis, apical domes may be more suitable as explants for transformation and subsequent regeneration of transclones than detached cotyledons.

Revue / Journal Title

Plant cell, tissue and organ culture    ISSN  0167-6857   CODEN PTCEDJ 

Source / Source

Congrès
New Zealand Regional IAPTC & B. Biennial Conference No14, Grand Chateau, Mount Ruapehu , NOUVELLE-ZELANDE (21/02/2001)
2002, vol. 70, no 1 (124 p.)  (1 P.1/4), pp. 51-60

Langue / Language

Anglais

Editeur / Publisher

Springer, Heidelberg, ALLEMAGNE  (1981) (Revue)

Mots-clés anglais / English Keywords

Plant pathogen

;

Softwood forest tree

;

Bacteria

;

Rhizobiaceae

;

Spermatophyta

;

Gymnospermae

;

Coniferales

;

Experimental protocol

;

Apex

;

Meristem

;

Cotyledon

;

Agrobacterium tumefaciens

;

Pinus radiata

;

Embryo

;

Gene expression

;

Stability

;

Under vacuum

;

Infiltration

;

Organogenesis

;

Tissue culture

;

Genetic transformation

;

Technique

;

Mots-clés français / French Keywords

Phytopathogène

;

Arbre forestier résineux

;

Bactérie

;

Rhizobiaceae

;

Spermatophyta

;

Gymnospermae

;

Coniferales

;

Protocole expérimental

;

Apex

;

Méristème

;

Cotylédon végétal

;

Agrobacterium tumefaciens

;

Pinus radiata

;

Embryon

;

Expression génique

;

Stabilité

;

Sous vide

;

Infiltration

;

Organogenèse

;

Culture tissu

;

Transformation génétique

;

Technique

;

Mots-clés espagnols / Spanish Keywords

Fitopatógeno

;

Arbol forestal resinoso

;

Bacteria

;

Rhizobiaceae

;

Spermatophyta

;

Gymnospermae

;

Coniferales

;

Protocolo experimental

;

Apex

;

Meristema

;

Cotiledón

;

Agrobacterium tumefaciens

;

Pinus radiata

;

Embrión

;

Expresión genética

;

Estabilidad

;

En vacío

;

Infiltración

;

Organogénesis

;

Cultivo tejido

;

Transformación genética

;

Técnica

;

Localisation / Location

INIST-CNRS, Cote INIST : 19374, 35400010440825.0060

Nº notice refdoc (ud4) : 13824106



Faire une nouvelle recherche
Make a new search
Lancer la recherche
Bas