Genetic and molecular analysis of Arabidopsis thaliana (ecotype 'Estland')transformed with Agrobacterium

Agrobacterium-mediated transformation of Arabidopsis, ecotype 'Estland', wa s established from root explants using kanamycin selection. Continuous ligh t during callus and shoot induction phases was promotive for shoot regenera tion, as compared to light/dark cycles. Use of optimized conditions for tra nsformation led to the formation of kanamycin-resistant calluses (up to 77% ) and transformed plantlets at a frequency of up to 45%. Southern analysis showed the presence of 1, 2, or more T-DNA inserts in 33%, 50%, and 17% of the primary transformants, respectively. Mendelian, as well as non-Mendelia n, inheritance patterns were obtained upon screening the progeny (TI) of va rious transformants for the expression of gus and nptII genes; the analysis of some of these transformants at the molecular level also corroborated th e Mendelian inheritance pattern. Moreover, genotypes of the T1 progeny coul d be predicted on the basis of T2 progeny analysis.