Articles
DEVELOPMENT AND GROWTH OF EMBRYOGENIC SUSPENSION CULTURES OF VITIS VINIFERA CVS IN BIOREACTOR AS A SYSTEM FOR GENETIC TRANSFORMATION
Article number
447_61
Pages
313 – 316
Language
Abstract
Suspension cultures in shake flasks were developed from embryogenic tissue cultures established from anthers of four commercial cultivars of Vitis vinifera L. [cv. "Superior seedless" (SR), cv. 49, cv. "Red Globe" (RG) and cv. 902]. We studied the suitability of this type of suspension culture for Agrobacterium mediated transformation.
The selection of a highly morphogenetic cell suspension resulted in obtaining a collection of 17 lines of embryogenic cell and embryoid like-suspension cultures of the above cultivars.
It was found that all these lines retain their high embryogenic ability, but they differed in the duration of culture on induction media.
The possibility of growing the suspension cultures of cvs. 902 and RG in the conical bubble type bioreactors with working volume up to 0.7 1 in batch regime, and several cycles of draw-fill regime was demonstrated over the course of 6 months cultivation.
High level and low fluctuation of embryogenic activity of these suspension cultures grown in the bioreactor was found.
Susceptibility to Agrobacterium mediated transformation of these cell suspension of cv.
RG from fourth and fifth cycles of bioreactor cultivation and cv. 902 obtained from a shake flask was studied by exposure to Agrobacterium tumefaciens harboring the uidA gene.
High efficiency of cytosolic GUS activity was determined by staining of the cells with X-gluc.
The selection of a highly morphogenetic cell suspension resulted in obtaining a collection of 17 lines of embryogenic cell and embryoid like-suspension cultures of the above cultivars.
It was found that all these lines retain their high embryogenic ability, but they differed in the duration of culture on induction media.
The possibility of growing the suspension cultures of cvs. 902 and RG in the conical bubble type bioreactors with working volume up to 0.7 1 in batch regime, and several cycles of draw-fill regime was demonstrated over the course of 6 months cultivation.
High level and low fluctuation of embryogenic activity of these suspension cultures grown in the bioreactor was found.
Susceptibility to Agrobacterium mediated transformation of these cell suspension of cv.
RG from fourth and fifth cycles of bioreactor cultivation and cv. 902 obtained from a shake flask was studied by exposure to Agrobacterium tumefaciens harboring the uidA gene.
High efficiency of cytosolic GUS activity was determined by staining of the cells with X-gluc.
Authors
A. Kh. Lipsky, N. Sahar, D. Holland, Moshe A. Flaishman, A. Perl
Keywords
embryogenic activity, blockage of development, scaling up, acetosyringone, GUS expression
Online Articles (132)