Abstract: A method of preparation of a plant tissue fragment is provided wherein apical dominance of plant meristematlc tissue is inhibited followed by fragmentation of the tissue. Also provided are methods of plant micropropagation and methods of artificial seed production using apical dominance suppression in preferably, a semi-automated process. Also provided is a plant tissue processing machine that generates plant fragments with high regeneration efficiency and an artificial seed production apparatus.

Abstract: A method of producing a transgenic monocotyledonous plant includes culturing a thin section explant from a monocotyledonous plant, such as sugarcane, wheat or sorghum, in the presence of an auxin and, optionally, a cytokinin, prior to transformation. Optimally, the thin section is oriented during this pre-transformation culture period of 1-6 days so that a basal surface is substantially not in contact with the culture medium. The cultured explant is then transformed followed by a rest period of 4-15 days in a culture medium without selection agent but comprising an auxin and, optionally, a cytokinin. After this rest period, transgenic plants are selectively propagated from the transformed plant tissue in the presence of a selection agent such as paromomycin sulphate or geneticin. This system provides rapid, efficient generation of transgenic monocotyledonous plants from transformed, non-callus tissue and thereby reduces the likelihood of somaclonal variation among transgenic progeny.

Abstract: A method of plant micopropagation is provided whereby a thin section of non-callus tissue obtained from a monocotyledonous plant, such as sugarcane, wheat or sorghum, is cultured in the presence of a cytokinin and/or an auxin. Optimal regeneration occurs when a basal surface of the thin section is oriented so as to be substantially not in contact with the culture medium. This micropropagation method produces mature monocotyledonous plants by either organogenic or embryogenic regeneration without substantial callus formation. By avoiding use of callus as a starting tissue, this method reduces the likelihood of propagated plants displaying somaclonal variation.