Patent ID: 9055721
Filing Date: 2015-06-16
Classification: A01H

Abstract:
1. A method of propagating a graminaceous plant, the method comprising the steps of: a) using immature inflorescence as explants to induce embryogenic calli, wherein step a) comprises washing, sterilizing, and culturing the immature inflorescence in callus induction medium comprising MS basal medium, MS vitamins, 20-35 g/l of sucrose, 1-4 mg/l of 2,4-D, 0.1-1.5 mg/l of BA, 500-4000 mg/l of proline, 0-2000 mg/l of MgCl b) performing callus multiplication in liquid suspension medium or on solid medium to propagate embryogenic calli, wherein step b) comprises culturing induced calli from step a) in a suspension culture medium comprising MS basal medium, MS vitamins, 30 g/l sucrose, 2 mg/l 2,4-D, 0.5 mg/l BA, 0.5-5.0 mg/l of ABA, 250-2000 mg/l of proline, and 100-150 mg/l of myo-inositol; c) regenerating shoots from the embryogenic calli of step b) on regeneration medium, wherein the regeneration medium comprises MS basal medium, vitamins, 30 g/l sucrose, 1-8 mg/l of BA, 0.1-1.0 mg/l of 2,4-D, 0.1-1.5 μM of GA, 500-2000 mg/l of proline, 0-750 mg/l of MgCl d) culturing the shoots in liquid shoot multiplication media for shoot multiplication resulting in an increased number of shoots, wherein the liquid shoot multiplication media comprises MS medium, 30 g/l of sucrose, 2.0-8.0 mg/l of BA, 0.1-2.0 mg/l of IAA, and 0-1.0 mg/l of IBA; e) culturing the shoots in liquid shoot quality improvement media for shoot quality improvement resulting in a further increased number of shoots and an increase in shoot diameter, wherein the liquid shoot quality improvement media comprises MS medium, vitamins, 1.0-8.0 mg/l of BA, 0-2.0 mg/l of IAA and 0-1.0 mg/l of IBA; and f) transferring the shoots to liquid rooting medium comprising activated charcoal to produce plantlets, wherein the liquid rooting medium comprises MS basal medium, MS vitamins and activated charcoal.