High efficiency initiation of regenerable embryogenic callus from anther filaments of 19-grapevine genotypes grown worldwide

The need for mastery in somatic cell culture and plant regeneration in the Grapevine, beyond its initial application for genetic transformation, has further increased when considering its potential use for deciphering the divergent genetic evolutions of somatic cells. In order to overcome the Grapevine’s recalcitrance to tissue culture, we developed an approach for the induction of embryogenic callus from anthers. Better-responding cells from anther’s filaments were identified enabling a callogenesis rate of nearly 100%. The optimisation of culture media composition, each one better adapted to the physiological stage of the cells, led to the design of three culture steps. The root stocks 3309C, 110R, 41B, Rupestris St. Georges, Fercal, SO4 and the Vitis vinifera genotypes Chardonnay, Chasselas blanc, Cabernet Sauvignon, Gewurztraminer, Grenache, Merlot, Muscat Ottonel, Pinot Meunier, Pinot Gris, Pinot Noir, Sauvignon, Syrah and five different clonal accessions of Riesling were amenable to reproducible efficient initiation of regenerable embryogenic callus. When combined with the developed regeneration procedure [1] we show that novel medium compositions and optimised culture steps offer a complete way leading from canes to embryogenic callus initiation and to subsequent plant regeneration.