Note : Les revendications sont présentées dans la langue officielle dans laquelle elles ont été soumises.
21
1. A manufactured seed comprising:
(a) a structure for containing a plant embryo; and
(b) a nutritive medium comprising about 5 mM to about 30 mM urea,
about 0.01 mM to about 8 mM L-arginine, and about 0.001 mM to
about 0.01 mM thiamine-HCl wherein the nutritive medium is
positioned to be capable of functional contact with at least a root end
of the plant embryo.
2. The manufactured seed according to claim 1, wherein the urea
concentration is about 5 mM to about 20 mM.
3. The manufactured seed according to claim 1 or 2, wherein the
urea concentration is about 13.32 mM.
4. The manufactured seed according to any one of claims 1 through
3, wherein the arginine concentration is about 0.01 mM to about 4 mM.
5. The manufactured seed according to any one of claims 1 through
4, wherein the arginine concentration is about 1.0471 mM.
6. The manufactured seed according to any one of claims 1 through
5, wherein the thiamine-HCl concentration is about 0.001 mM to about 0.005
mM.
7. The manufactured seed according to any one of claims 1 through
6, wherein the thiamine-HCl concentration is about 0.003 mM.
8. The manufactured seed according to any one of claims 1 through
7, wherein the nutritive medium further comprises charcoal.
22
9. The manufactured seed according to claim 8, wherein the
charcoal concentration is about 2 g/L to 5 g/L.
10. The manufactured seed according to claim 9, wherein the
charcoal concentration is about 2 g/L to 3 g/L.
11. The manufactured seed according to claim 10, wherein the
charcoal concentration is about 2.5 g/L.
12. The manufactured seed according to any one of claims 1 through
11, wherein the nutritive medium further comprises a non-protein amino acid.
13. The manufactured seed according to claim 12, wherein the non-
protein amino acid concentration is about 0.1 mM to 3 mM.
14. The manufactured seed according to claim 13, wherein the non-
protein amino acid concentration is about 0.1 mM to 1.25 mM.
15. The manufactured seed according to any one of claims 12
through 14, wherein the non-protein amino acid is ornithine.
16. The manufactured seed according to any one of claims 12
through 14, wherein the non-protein amino acid is selected from the group
consisting of arginosuccinate, citrulline, and canavanine.
17. The manufactured seed according to any one of claims 1 through
16, wherein the nutritive medium further comprises sucrose.
18. The manufactured seed according to claim 17, wherein the
sucrose concentration is about 146.07 mM.
23
19. The manufactured seed according to claim 17, wherein the
sucrose concentration is about 58.43 g/L.
20. The manufactured seed according to any one of claims 1 through
19, wherein the nutritive medium further comprises a smoke suspension.
21. The manufactured seed according to claim 20, wherein the smoke
suspension concentration is about 0.1 mL/L medium to 10 mL/L medium.
22. The manufactured seed according to claim 21, wherein the smoke
suspension concentration is about 0.1 mL/L medium to 1 mL/L medium.
23. The manufactured seed according to any one of claims 1 through
22, wherein the nutritive medium further comprises a polyamine selected from
the group consisting of putrescine, spermidine, spermine, and mixtures
thereof.
24. The manufactured seed according to claim 23, wherein the
polyamine concentration is about 0.1 mM to 0.5 mM.
25. The manufactured seed according to claim 1, wherein the
nutritive medium further comprises at least one component selected from the
group consisting of about 0.01 mM to 2 mM L-asparagine, 0.01 mM to 6 mM L-
glutamine, 0.01 mM to 2 mM L-lysine, 0.01 mM to 2 mM L-serine, 0.01 mM to
2 mM L-proline, 0.01 mM to 2 mM L-valine, 0.01 mM to 2 mM L-alanine, 0.01
mM to 2 mM L-cysteine, 0.01 mM to 2 mM L-leucine, 0.01 mM to 2 mM L-
tyrosine, 0.01 mM to 2 mM L-threonine, 0.01 mM to 2 mM L-phenylalanine,
0.01 mM to 2 mM L-histidine, 0.002 mM to 0.2 mM L-glycine, 0.01 mM to 2
mM L-tryptophan, 0.01 mM to 2 mM L-isoleucine and 0.01 mM to 2 mM L-
methionine.
24
26. The manufactured seed according to claim 1, wherein the
nutritive medium further comprises at least one component selected from the
group consisting of about 0.01 mM to 1 mM L-asparagine, 0.01 mM to 3 mM L-
g1 utamine, 0.01 mM to 1 mM L-lysine, 0.01 mM to 1 mM L-serine, 0.01 mM to
1 mM L-proline, 0.01 mM to 1 mM L-valine, 0.01 mM to 1 mM L-alanine, 0.01
mM to 1 mM L-cysteine, 0.01 mM to 1 mM L-leucine, 0.01 mM to 1 mM L-
tyrosine, 0.01 mM to I mM L-threonine, 0.01 mM to 1 mM L-phenylalanine,
0.01 mM to 1 mM L-histidine, 0.01 mM to 0.1 mM L-glycine, 0.01 mM to 1 mM
L-tryptophan, 0.01 mM to 1 mM L-isoleucine and 0.01 mM to 1 mM L-
methionine.
27. The manufactured seed according to claim 1wherein the nutritive
medium further comprises at least one component selected from the group
consisting of about 0.8076 mM L-asparagine, 1.8248 mM L-glutamine, 0.1624
mM L-lysine, 0.7613 mM L-serine, 0.463 mM L-proline, 0.455 mM L-valine,
0.5983 mM L-alanine, 0.2204 mM L-cysteine, 0.6099 mM L-leucine, 0.2942
mM L-tyrosine, 0.2241 mM L-threonine, 0.3227 mM L-phenylalanine, 0.1721
mM L-histidine, 0.71 mM L-glycine, 0.1307 mM L-tryptophan, 0.2036 mM L-
isoleucine, and 0.1789 mM L-methionine.
28. The manufactured seed according to claim 1, wherein the
nutritive medium further comprises at least one component selected from the
group consisting of about 10 g/L to 30 g/L gel solute, 5 mM to 200 mM carbon
source, 10 mM to 20 mM KNO3, 2.5 mM to 5 mM NH4NO3, 0.002 mM to 0.01
mM KI, 0.01 mM to 20 mM KH2PO4, 0.5 mM to 5 mM CaCl2, 0.1 mM to 10 mM
MgSO4, 0.01 mM to 0.1 mM Na2EDTA, 0.01 mM to 0.1 mM FeSO4, 0.1 mM to
0.5 mM ferric citrate, 0.02 mM to 0.1 mM MnSO4, 0.01 mM to 0.1 mM MnCl2,
0.01 mM to 1 mM H3BO3, 0.0001 mM to 0.005 mM ZnSO4, 1x10 -5 mM to 1x10-
4 mM CoCl2, 0.001 mM to 0.005 mM CuCl2, 1x10 -5 mM to 1x10 -4 mM CuSO4,
1x10 -4 mM to 0.001 mM Na2MoO4, 1x10 -4 mM to 0.001 mM (NH4)2MoO4, 1x10-
4 mM to 0.001 mM Riboflavin, 0.001 mM to 0.005mM pyridoxine.cndot.HCl, 0.0005
25
mM to 0.004 mM Ca-pantothenate, 0.001 mM to 0.02 mM nicotinic acid, 1x10 -7
mM to 1x10 -5 mM Biotin, 1x10 -4 mM to 1x10 -3 mM folic acid, and 0.1 mM to
2.5
mM myo-inositol.
29. The manufactured seed according to claim 1, wherein the
nutritive medium further comprises at least one component selected from the
group consisting of about 10 g/L to 25 g/L gel solute, 25 mM to 175 mM
carbon source, 10 mM to 15 mM KNO3, 2 mM to 4 mM NH4NO3, 0.002 mM to
0.005 mM KI, 0.05 mM to 18 mM KH2PO4, 1 mM to 3 mM CaCl2, 0.2 mM to 3
mM MgSO4, 0.01 mM to 0.8 mM Na2EDTA, 0.01 mM to 0.05 mM FeSO4, 0.1
mM to 0.4 mM ferric citrate, 0.02 mM to 0.5 mM MnSO4, 0.01 mM to 0.05 mM
MnCl2, 0.1 mM to 0.5 mM H3BO3, 0.0003 mM to 0.004 mM ZnSO4, 0.001 mM
to 0.004 mM pyridoxine-HCl, 0.0005 mM to 0.015 mM Ca-pantothenate, 0.001
mM to 0.01 mM nicotinic acid, and 0.1 mM to 1 mM myo-inositol.
30. The manufactured seed according to claim 1, wherein the
nutritive medium further comprises at least one component selected from the
group consisting of about 3.74 mM NH4NO3, 13.23 mM KH2PO4, 1.54 mM
CaCl2, 1.98 mM MgSO4, 0.245 mM ferric citrate, 0.019 mM MnCl2, 0.162 mM
H3B03, 0.0023 mM CuCl2, 0.0003 mM (NH4)2MoO4, 0.1 mg/L GA4/7, 0.0003
mM Riboflavin, 0.0012 mM pyridoxine.cndot.HCl, 0.001 mM Ca-pantothenate,
0.0081 mM nicotinic acid, 4.09x10 -6 Biotin, 0.0003 mM folic acid, and 0.555
mM myo-inositol.
31. The manufactured seed according to claim 1, wherein the
nutritive medium further comprises at least one component selected from the
group consisting of about 11.57 mM KNO3, 2.577 mM NH4NO3, 0.0025 mM KI,
0.625 mM KH2PO4, 1.133 mM CaCl2, 0.367 mM MgSO4, 0.05 mM Na2EDTA,
0.027 mM FeSO4, 0.045 mM MnSO4, 0.05 mM H3BO3, 8.4x10 -3 mM ZnSO4,
3.2x10 -5 mM CuSO4, 2.9x10 -5 mM CoCl2, 4.4x10 -4 mM Na2MoO4, 0.0024 mM
26
pyridoxine.cndot.HCl, 0.0041 mM nicotinic acid, 0.555 mM myo-inositol, and
0.027
mM L-glycine.
32. The manufactured seed according to any one of claims 1 through
31, wherein the nutritive medium further comprises at least one component
selected from the group consisting of about 18 g/L agar, 3.6 g/L Pluronic F-
68,
and 240 mL/L DC-200 silicone oil.
33. The manufactured seed according to any one of claims 1 through
32, wherein the pH of nutritive medium is about 5.7.
34. The manufactured seed according to any one of claims 1 through
33 wherein the plant embryo is a gymnosperm plant embryo.
35. The manufactured seed according to claim 34 wherein the
gymnosperm is Pine.
36. A method for germinating a plant embryo, comprising placing at
least a root end the plant embryo in functional contact with a nutritive
medium
comprising about 5 mM to about 30 mM urea, about 0.01 mM to about 8 mM
L-arginine, and about 0.001 mM to about 0.01 mM thiamine-HCl in a
manufactured seed.
37. The method according to claim 36, wherein the urea
concentration is about 5 mM to about 20 mM.
38. The method according to claim 36 or 37, wherein the urea
concentration is about 13.32 mM.
39. The method according to any one of claims 36 through 38,
wherein the arginine concentration is about 0.01 mM to about 4 mM.
27
40. The method according to any one of claims 36 through 39,
wherein the arginine concentration is about 1.0471 mM.
41. The method according to any one of claims 36 through 40,
wherein the thiamine-HCl concentration is about 0.001 mM to about 0.005
mM.
42. The method according to any one of claims 36 through 41,
wherein the thiamine-HCl concentration is about 0.003 mM.
43. The method according to any one of claims 36 through 42,
wherein the nutritive medium further comprises charcoal.
44. The method according to claim 43, wherein the charcoal
concentration is about 2 g/L to 5 g/L.
45. The method according to claim 44, wherein the charcoal
concentration is about 2 g/L to 3 g/L.
46. The method according to claim 45, wherein the charcoal
concentration is about 2.5 g/L.
47. The method according to any one of claims 36 through 46,
wherein the nutritive medium further comprises a non-protein amino acid.
48. The method according to claim 47, wherein the non-protein amino
acid concentration is about 0.1 mM to 3 mM.
49. The method according to claim 48, wherein the non-protein amino
acid concentration is about 0.1 mM to 1.25 mM.
28
50. The method according to any one of claims 47 through 49,
wherein the non-protein amino acid is ornithine.
51. The method according to any one of claims 47 through 49,
wherein the non-protein amino acid is selected from the group consisting of
arginosuccinate, citrulline, and canavanine.
52. The method according to any one of claims 36 through 51,
wherein the nutritive medium further comprises sucrose.
53. The method according to claim 52, wherein the sucrose
concentration is about 146.07 mM.
54. The method according to claim 52, wherein the sucrose
concentration is about 58.43 g/L.
55. The method according to any one of claims 36 through 54,
wherein the nutritive medium further comprises a smoke suspension.
56. The method according to claim 55, wherein the smoke
suspension concentration is about 0.1 mL/L medium to 10 mL/L medium.
57. The method according to claim 56, wherein the smoke
suspension concentration is about 0.1 mL/L medium to 1 mL/L medium.
58. The method according to any one of claims 36 through 57,
wherein the nutritive medium further comprises a polyamine selected from the
group consisting of putrescine, spermidine, spermine, and mixtures thereof.
59. The method according to claim 58, wherein the polyamine
concentration is about 0.1 mM to 0.5 mM.
29
60. The method according to claim 36, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 0.01 mM to 2 mM L-asparagine, 0.01 mM to 6 mM L-glutamine, 0.01
mM to 2 mM L-lysine, 0.01 mM to 2 mM L-serine, 0.01 mM to 2 mM L-proline,
0.01 mM to 2 mM L-valine, 0.01 mM to 2 mM L-alanine, 0.01 mM to 2 mM L-
cysteine, 0.01 mM to 2 mM L-leucine, 0.01 mM to 2 mM L-tyrosine, 0.01 mM
to 2 mM L-threonine, 0.01 mM to 2 mM L-phenylalanine, 0.01 mM to 2 mM L-
histidine, 0.002 mM to 0.2 mM L-glycine, 0.01 mM to 2 mM L-tryptophan, 0.01
mM to 2 mM L-isoleucine and 0.01 mM to 2 mM L-methionine
61. The method according to claim 36, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 0.01 mM to 1 mM L-asparagine, 0.01 mM to 3 mM L-glutamine, 0.01
mM to 1 mM L-lysine, 0.01 mM to 1 mM L-serine, 0.01 mM to 1 mM L-proline,
0.01 mM to 1 mM L-valine, 0.01 mM to 1 mM L-alanine, 0.01 mM to 1 mM L-
cysteine, 0.01 mM to 1 mM L-leucine, 0.01 mM to 1 mM L-tyrosine, 0.01 mM
to I mM L-threonine, 0.01 mM to 1 mM L-phenylalanine, 0.01 mM to 1 mM L-
histidine, 0.01 mM to 0.1 mM L-glycine, 0.01 mM to 1 mM L-tryptophan, 0.01
mM to 1 mM L-isoleucine and 0.01 mM to 1 mM L-methionine.
62. The method according to claim 36, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 0.8076 mM L-asparagine, 1.8248 mM L-glutamine, 0.1624 mM L-
lysine, 0.7613 mM L-serine, 0.463 mM L-proline, 0.455 mM L-valine, 0.5983
mM L-alanine, 0.2204 mM L-cysteine, 0.6099 mM L-leucine, 0.2942 mM L-
tyrosine, 0.2241 mM L-threonine, 0.3227 mM L-phenylalanine, 0.1721 mM L-
histidine, 0.71 mM L-glycine, 0.1307 mM L-tryptophan, 0.2036 mM L-
isoleucine, and 0.1789 mM L-methionine.
63. The method according to claim 36, wherein the nutritive medium
further comprises at least one component selected from the group consisting
30
of about 10 g/L to 30 g/L gel solute, 5 mM to 200 mM carbon source, 10 mM
to 20 mM KNO3, 2.5 mM to 5 mM NH4NO3, 0.002 mM to 0.01 mM KI, 0.01 mM
to 20 mM KH2PO4, 0.5 mM to 5 mM CaCl2, 0.1 mM to 10 mM MgSO4, 0.01 mM
to 0.1 mM Na2EDTA, 0.01 mM to 0.1 mM FeSO4, 0.1 mM to 0.5 mM ferric
citrate, 0.02 mM to 0.1 mM MnSO4, 0.01 mM to 0.1 mM MnCl2, 0.01 mM to 1
mM H3BO3, 0.0001 mM to 0.005 mM ZnSO4, 1x10 -5 mM to 1x10 -4 mM CoCl2,
0.001 mM to 0.005 mM CuCl2, 1x10 -5 mM to 1x10 -4 mM CuSO4, 1x10 -4 mM to
0.001 mM Na2MoO4, 1x10 -4 mM to 0.001 mM (NH4)2MoO4, 1x10 -4 mM to 0.001
mM Riboflavin, 0.001 mM to 0.005mM pyridoxine.cndot.HCl, 0.0005 mM to 0.004
mM Ca-pantothenate, 0.001 mM to 0.02 mM nicotinic acid, 1x10 -7 mM to 1x10 -
mM Biotin, 1x10 -4 mM to 1x10 -3 mM folic acid, and 0.1 mM to 2.5 mM myo-
inositol.
64. The method according to claim 36, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 10 g/L to 25 g/L gel solute, 25 mM to 175 mM carbon source, 10 mM
to 15 mM KNO3, 2 mM to 4 mM NH4NO3, 0.002 mM to 0.005 mM KI, 0.05 mM
to 18 mM KH2PO4, 1 mM to 3 mM CaCl2, 0.2 mM to 3 mM MgSO4, 0.01 mM to
0.8 mM Na2EDTA, 0.01 mM to 0.05 mM FeSO4, 0.1 mM to 0.4 mM ferric
citrate, 0.02 mM to 0.5 mM MnSO4, 0.01 mM to 0.05 mM MnCl2, 0.1 mM to 0.5
mM H3BO3, 0.0003 mM to 0.004 mM ZnSO4, 0.001 mM to 0.004 mM
pyridoxine.cndot.HCl, 0.0005 mM to 0.015 mM Ca-pantothenate, 0.001 mM to 0.01
mM nicotinic acid, and 0.1 mM to 1 mM myo-inositol.
65. The method according to claim 36, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 3.74 mM NH4NO3, 13.23 mM KH2PO4, 1.54 mM CaCl2, 1.98 mM
MgSO4, 0.245 mM ferric citrate, 0.019 mM MnCl2, 0.162 mM H3BO3, 0.0023
mM CuCl2, 0.0003 mM (NH4)2MoO4, 0.1 mg/L GA4/7, 0.0003 mM Riboflavin,
0.0012 mM pyridoxine.cndot.HCl, 0.001 mM Ca-pantothenate, 0.0081 mM nicotinic
acid, 4.09x10 -6 Biotin, 0.0003 mM folic acid, and 0.555 mM myo-inositol.
31
66. The method according to claim 36, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 11.57 mM KNO3, 2.577 mM NH4NO3, 0.0025 mM KI, 0.625 mM
KH2PO4, 1.133 mM CaCl2, 0.367 mM MgSO4, 0.05 mM Na2EDTA, 0.027 mM
FeSO4, 0.045 mM MnSO4, 0.05 mM H3B03, 8.4x10 -3 mM ZnSO4, 3.2x10 -5 mM
CuSO4, 2.9x10 -5 mM CoCl2, 4.4x10 -4 mM Na2MoO4, 0.0024 mM
pyridoxine.cndot.HCl, 0.0041 mM nicotinic acid, 0.555 mM myo-inositol, and
0.027
mM L-glycine.
67. The method according to any one of claims 36 through 66,
wherein the nutritive medium further comprises at least one component
selected from the group consisting of about 18 g/L agar, 3.6 g/L Pluronic F-
68,
and 240 mL/L DC-200 silicone oil.
68. The method according to any one of claims 36 through 67,
wherein the pH of nutritive medium is about 5.7.
69. A method for germinating a plant embryo comprising placing the plant
embryo in a manufactured seed according to any one of claims 1 through 35.
70. The use of a nutritive medium for germinating a plant embryo by
placing the nutritive medium into functional contact with at least a root end
of the
plant embryo, the nutritive medium comprising about 5 mM to about 30 mM urea,
about 0.01 mM to about 8 mM L-arginine, and about 0.001 mM to about 0.01 mM
thiamine-HCl in a manufactured seed.
71. The use according to claim 70, wherein the urea concentration is
about 5 mM to about 20 mM.
72. The use according to claim 70 or 71, wherein the urea
concentration is about 13.32 mM.
32
73. The use according to any one of claims 70 through 72, wherein
the arginine concentration is about 0.01 mM to about 4 mM.
74. The use according to any one of claims 70 through 73, wherein
the arginine concentration is about 1.0471 mM.
75. The use according to any one of claims 70 through 74, wherein
the thiamine-HCl concentration is about 0.001 mM to about 0.005 mM.
76. The use according to any one of claims 70 through 75, wherein
the thiamine-HCl concentration is about 0.003 mM.
77. The use according to any one of claims 70 through 76, wherein
the nutritive medium further comprises charcoal.
78. The use according to claim 77, wherein the charcoal
concentration is about 2 g/L to 5 g/L.
79. The use according to claim 78, wherein the charcoal
concentration is about 2 g/L to 3 g/L.
80. The use according to claim 79, wherein the charcoal
concentration is about 2.5 g/L.
81. The use according to any one of claims 70 through 80, wherein
the nutritive medium further comprises a non-protein amino acid.
82. The use according to claim 81, wherein the non-protein amino
acid concentration is about 0.1 mM to 3 mM.
33
83. The use according to claim 82, wherein the non-protein amino
acid concentration is about 0.1 mM to 1.25 mM.
84. The use according to any one of claims 81 through 83, wherein
the non-protein amino acid is ornithine.
85. The use according to any one of claims 81 through 83, wherein
the non-protein amino acid is selected from the group consisting of
arginosuccinate, citrulline, and canavanine.
86. The use according to any one of claims 70 through 85, wherein
the nutritive medium further comprises sucrose.
87. The use according to claim 86, wherein the sucrose concentration
is about 146.07 mM.
88. The use according to claim 86, wherein the sucrose concentration
is about 58.43 g/L.
89. The use according to any one of claims 70 through 88, wherein
the nutritive medium further comprises a smoke suspension.
90. The use according to claim 189, wherein the smoke suspension
concentration is about 0.1 mL/L medium to 10 mL/L medium.
91. The use according to claim 90, wherein the smoke suspension
concentration is about 0.1 mL/L medium to 1 mL/L medium.
92. The use according to any one of claims 70 through 91, wherein
the nutritive medium further comprises a polyamine selected from the group
consisting of putrescine, spermidine, spermine, and mixtures thereof.
34
93. The use according to claim 92, wherein the polyamine
concentration is about 0.1 mM to 0.5 mM.
94. The use according to claim 70, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 0.01 mM to 2 mM L-asparagine, 0.01 mM to 6 mM L-glutamine, 0.01
mM to 2 mM L-lysine, 0.01 mM to 2 mM L-serine, 0.01 mM to 2 mM L-proline,
0.01 mM to 2 mM L-valine, 0.01 mM to 2 mM L-alanine, 0.01 mM to 2 mM L-
cysteine, 0.01 mM to 2 mM L-leucine, 0.01 mM to 2 mM L-tyrosine, 0.01 mM
to 2 mM L-threonine, 0.01 mM to 2 mM L-phenylalanine, 0.01 mM to 2 mM L-
histidine, 0.002 mM to 0.2 mM L-glycine, 0.01 mM to 2 mM L-tryptophan, 0.01
mM to 2 mM L-isoleucine and 0.01 mM to 2 mM L-methionine.
95. The use according to claim 70, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 0.01 mM to 1 mM L-asparagine, 0.01 mM to 3 mM L-glutamine, 0.01
mM to 1 mM L-lysine, 0.01 mM to 1 mM L-serine, 0.01 mM to 1 mM L-proline,
0.01 mM to 1 mM L-valine, 0.01 mM to 1 mM L-alanine, 0.01 mM to 1 mM L-
cysteine, 0.01 mM to 1 mM L-leucine, 0.01 mM to 1 mM L-tyrosine, 0.01 mM
to I mM L-threonine, 0.01 mM to 1 mM L-phenylalanine, 0.01 mM to 1 mM L-
histidine, 0.01 mM to 0.1 mM L-glycine, 0.01 mM to 1 mM L-tryptophan, 0.01
mM to 1 mM L-isoleucine and 0.01 mM to 1 mM L-methionine.
96. The use according to claim 70, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 0.8076 mM L-asparagine, 1.8248 mM L-glutamine, 0.1624 mM L-
lysine, 0.7613 mM L-serine, 0.463 mM L-proline, 0.455 mM L-valine, 0.5983
mM L-alanine, 0.2204 mM L-cysteine, 0.6099 mM L-leucine, 0.2942 mM L-
tyrosine, 0.2241 mM L-threonine, 0.3227 mM L-phenylalanine, 0.1721 mM L-
histidine, 0.71 mM L-glycine, 0.1307 mM L-tryptophan, 0.2036 mM L-
isoleucine, and 0.1789 mM L-methionine.
35
97. The use according to claim 70, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 10 g/L to 30 g/L gel solute, 5 mM to 200 mM carbon source, 10 mM
to 20 mM KNO3, 2.5 mM to 5 mM NH4NO3, 0.002 mM to 0.01 mM KI, 0.01 mM
to 20 mM KH2PO4, 0.5 mM to 5 mM CaCl2, 0.1 mM to 10 mM MgSO4, 0.01 mM
to 0.1 mM Na2EDTA, 0.01 mM to 0.1 mM FeSO4, 0.1 mM to 0.5 mM ferric
citrate, 0.02 mM to 0.1 mM MnSO4, 0.01 mM to 0.1 mM MnCl2, 0.01 mM to 1
mM H3B03, 0.0001 mM to 0.005 mM ZnSO4, 1x10 -5 mM to 1x10 -4 mM CoCl2,
0.001 mM to 0.005 mM CuCl2, 1x10 -5 mM to 1x10 -4 mM CuSO4, 1x10 -4 mM to
0.001 mM Na2MoO4, 1x10 -4 mM to 0.001 mM (NH4)2MoO4, 1x10 -4 mM to 0.001
mM Riboflavin, 0.001 mM to 0.005mM pyridoxine.cndot.HCl, 0.0005 mM to 0.004
mM Ca-pantothenate, 0.001 mM to 0.02 mM nicotinic acid, 1x10 -7 mM to 1x10 -
mM Biotin, 1x10 -4 mM to 1x10 -3 mM folic acid, and 0.1 mM to 2.5 mM myo-
inositol.
98. The use according to claim 70, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 10 g/L to 25 g/L gel solute, 25 mM to 175 mM carbon source, 10 mM
to 15 mM KNO3, 2 mM to 4 mM NH4NO3, 0.002 mM to 0.005 mM KI, 0.05 mM
to 18 mM KH2PO4, 1 mM to 3 mM CaCl2, 0.2 mM to 3 mM MgSO4, 0.01 mM to
0.8 mM Na2EDTA, 0.01 mM to 0.05 mM FeSO4, 0.1 mM to 0.4 mM ferric
citrate, 0.02 mM to 0.5 mM MnSO4, 0.01 mM to 0.05 mM MnCl2, 0.1 mM to 0.5
mM H3BO3, 0.0003 mM to 0.004 mM ZnSO4, 0.001 mM to 0.004 mM
pyridoxine.cndot.HCl, 0.0005 mM to 0.015 mM Ca-pantothenate, 0.001 mM to 0.01
mM nicotinic acid, and 0.1 mM to 1 mM myo-inositol.
99. The use according to claim 70, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 3.74 mM NH4NO3, 13.23 mM KH2PO4, 1.54 mM CaCl2, 1.98 mM
MgSO4, 0.245 mM ferric citrate, 0.019 mM MnCl2, 0.162 mM H3BO3, 0.0023
mM CuCl2, 0.0003 mM (NH4)2MoO4, 0.1 mg/L GA4/7, 0.0003 mM Riboflavin,
36
0.0012 mM pyridoxine.cndot.HCl, 0.001 mM Ca-pantothenate, 0.0081 mM nicotinic
acid, 4.09x10 -6 Biotin, 0.0003 mM folic acid, and 0.555 mM myo-inositol.
100. The use according to claim 70, wherein the nutritive medium
further comprises at least one component selected from the group consisting
of about 11.57 mM KNO3, 2.577 mM NH4NO3, 0.0025 mM KI, 0.625 mM
KH2PO4, 1.133 mM CaCl2, 0.367 mM MgSO4, 0.05 mM Na2EDTA, 0.027 mM
FeSO4, 0.045 mM MnSO4, 0.05 mM H3BO3, 8.4x10 -3 mM ZnSO4, 3.2x10 -5 mM
CuSO4, 2.9x10 -5 mM CoCl2, 4.4x10 -4 mM Na2MoO4, 0.0024 mM
pyridoxine.cndot.HCl, 0.0041 mM nicotinic acid, 0.555 mM myo-inositol, and
0.027
mM L-glycine.
101. The use according to any one of claims 70 through 100, wherein
the nutritive medium further comprises at least one component selected from
the group consisting of about 18 g/L agar, 3.6 g/L Pluronic F-68, and 240 mL/L
DC-200 silicone oil.
102. The use according to any one of claims 70 through 101, wherein
the pH of nutritive medium is about 5.7.
103. The use according to any one of claims 70 through 102 wherein
the plant embryo is a gymnosperm plant embryo.
104. The use according to claim 103 wherein the gymnosperm is Pine.