Patent ID: 7368631

Claim:
A method of making transgenic Acacia mangium plants comprising: a) culturing stem pieces of Acacia mangium cells on culture medium comprising basic MS medium supplemented with a) thidiazuron, b) indole-3-acetic acid, c) casein enzymatic hydrolysate, d) L-ascorbic acid, e) L-glutamine, f) L-asparagine, g) L-proline, h) sucrose and i) phytagel or agar; b) co-cultivating the stem pieces of step (a) with Agrobacterium in medium comprising basic MS medium supplemented with a) thidiazuron, b) indole-3-acetic acid, c) casein enzymatic hydrolysate, d) L-ascorbic acid, e) L-glutamine, f) L-asparagine, g) L-proline, h) sucrose, i) acetosyringone and j) phytagel or agar to form infected stem pieces; c) culturing the infected stem pieces on medium comprising basic MS medium supplemented with a) thidiazuron, b) indole-3-acetic acid, c) casein enzymatic hydrolysate, d) L-ascorbic acid, e) L-glutamine, f) L-asparagine, g) L-proline, h) sucrose, i) an Agrobacterium eradicant, j) an antibiotic and k) phytagel or agar to induce and select transformed callus and adventitious bud formation on the stem pieces; d) culturing the stem pieces having callus and adventitious buds on a medium comprising basic MS medium supplemented with a) thidiazuron, b) indole-3-acetic acid, c) casein enzymatic hydrolysate, d) L-ascorbic acid, e) L-glutamine, f) L-asparagine, g) L-proline, h) sucrose, i) an antibiotic, j) gibberellic acid and k) phytagel or agar to promote adventitious bud elongation; e) culturing the elongated adventitious buds on medium comprising basic MS medium supplemented with a) 6-benzylaminopurine, b) gibberellic acid, c) casein enzymatic hydrolysate, d) L-ascorbic acid, e) L-glutamine, f) L-asparagine, g) L-proline, h) sucrose, and i) phytagel or agar to promote pinnate leaf formation on the adventitious buds; and f) culturing the adventitious buds with leaves on medium comprising ½ basic MS medium supplemented with a) α-naphthaleneacetic acid, b) kinetin, c) casein enzymatic hydrolysate, d) L-ascorbic acid, e) L-glutamine, f) L-asparagine, g) L-proline, h) sucrose, and i) phytagel to form rooted transgenic plantlets.