Obtainment Of Transgenic Wheat With The Insecticidal Lectin From Pinellia Pedatisecta Schott (Pinellia Pedatisecta Agglutinin; PPA) Gene

In recent years, the world witnessed the rapid development of plant transformation technology. Genetically modified cultivars from soybean and cotton have been widely grown and provided enormous economic and social benefits. Wheat is one of the major food crops for the world's population. The first report of wheat transgenic plants in 1992, provides a solid foundation for molecular breeding in wheat. The topic for purpose of solve the issues existing wheat transformation, we used PPA gene into the wheat and build a more mature transformation system for aphid resistance wheat.The callus of the immature embryos chosen from wheat rarieties,"shi4185"and"yangmai12"etc, were cultured to study the effect of various factors on callus induction and differentiation. And study the transformation efficiency on main factors of the particle bombardment. Plasmid which containing selectable PPA gene and GUS reporter gene, was directly delivered into immature embryos of wheat. hygromycin-resistant plants were selected by hygromycin and regenerated.The results showed that the best acceptors was embryonic callus of immature embryo. The callus placed in medium containing 0.4 mol?L-1 mannitol prior 6 hour and after 18 h to bombardment, which increase conversion rate of bombardment. The transient expression of GUS reached the highest when the callus of immature embryos were bombarded at the pressure of 1 100 psi, using 300μg gold particles and 1μg plasmid per bombardment. Bombarded callus cultured in the recovery medium containing 2 mg?L-1 2,4-D on the recovery 5 d-7 d can improve the plant regeneration capacity of callus. Resistant callus obtained after the 60 mg?L-1 hygromycin, resistant callus tested and stable GUS gene expression. And then differentiate green buds in differentiation medium contains KT 1 mg?L-1 and NAA 0.1 mg?L-1-0.5 mg?L-1, and induced seedling.