MONOCOTYLEDON TRANSGENIC METHOD FOR INVADING GROWING POINTS OF SEED BUDS MINIMALLY AND FULLY

摘要

The present invention is a method of shoot apical meristem transformation for monocot plant via sufficient and micro wounding (SMW). The technical process includes: expose the apical meristem by removing the coleoptile away when the shoot grows to 0.2-2 cm after 1-2 days of seed germination; make sufficient and micro wounding transformation to the apical meristem by stabbing and brushing for 2-3 times using the SMW brush having 100-5000 bristles which is 4-20 μm in diameter for each one and 0.5-3 mm in exposed length, and dipped with the Agrobacterium tumefaciens containing binary vector harboring exogenous genes; develop the treated meristems directly to normal plants after co-cultivation; promote the plants to develop big spikes and set more seeds; harvest the seeds of T₀ plants separately; detect and identify the transformation results in T₁ generation which is bred from each individual T₀ plant. The advantages of the invention are independent of tissue culture, unlimited in genotype, unnecessary to carry resistant marker, simple and large scale to perform, and applicable to all monocot plants which can set seeds. The transformation efficiencies for wheat, rice and maize using this method are 49%, 66.3%, and 100%, respectively.