%0 Journal Article
%9 ACL : Articles dans des revues avec comité de lecture répertoriées par l'AERES
%A Jiang, Q. B.
%A Ma, Y. Z.
%A Zhong, C. L.
%A Zeng, B. S.
%A Zhang, Y.
%A Pinyopusarerk, K.
%A Bogusz, Didier
%A Franche, Claudine
%T Optimization of the conditions for Casuarina cunninghamiana Miq. genetic transformation mediated by Agrobacterium tumefaciens
%D 2015
%L fdi:010064063
%G ENG
%J Plant Cell Tissue and Organ Culture
%@ 0167-6857
%K Actinorhizal plant ; Casuarina cunninghamiana ; Agrobacterium tumefaciens ; Genetic transformation ; Frankia
%M ISI:000351524900017
%N 1
%P 195-204
%R 10.1007/s11240-014-0694-y
%U https://www.documentation.ird.fr/hor/fdi:010064063
%> https://www.documentation.ird.fr/intranet/publi/2015/04/010064063.pdf
%V 121
%W Horizon (IRD)
%X Using epicotyl fragments of the actinorhizal tree Casuarina cunninghamiana and the disarmed strain of Agrobacterium tumefaciens C58C1 (pGV2260) containing the pBIN19-35S-GUSINT binary vector, a method for the genetic transformation of C. cunninghamiana was established. Transformed cells were initially selected for 2 weeks on nutrient medium supplemented with kanamycin 20 mg L-1 during callus induction, and then subcultured with 50 mg L-1 until adventitious bud and shoot differentiation. Different factors involved in the early stages of the T-DNA transfer process were studied. Agrobacterium-mediated DNA delivery was most successful when epicotyl fragments excised from 45-day-old seedlings were co-cultivated with an exponentially growing culture of A. tumefaciens at an OD600nm of 0.3, for 5 days in the presence of 50 mu M of acetosyringone. Kanamycin resistant calli were observed on 88.89 % of the explants and transgenic rooted C. cunninghamiana plants were obtained in 6 months. Evidence of genetic transformation was demonstrated by -glucuronidase histochemical assays and polymerase chain reaction analyses. The possibility to obtain transgenic nitrogen-fixing nodules after inoculation by the soil actinomycete Frankia was established.
%$ 084 ; 076