Somatic embryogemic regeneration of Acacia mangium
A technique for somatic embryos and explants, used in, preparing somatic embryos of Acacia mangium, regenerating plants or regenerating bodies, amang, regenerating Acacia mangium by somatic embryogenesis, sand-soil mixture, cloning and planting stock, mature culture Continue to cultivate on the base and germination medium, and grow into the field of embryogenic tissue, which can solve the problems of unstable plant tissue and abnormal type.
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Embodiment 1
[0025] Example 1: Reagents and media
[0026] name
molecular formula
concentration
(mg / l)
Sigma
catalog number
Macronutrients
NH 4 NO 3
1650
A-3795
KNO 3
1900
P-8291
CaCl 2 .2H 2 o
440
C-2536
Magnesium Sulfate Heptahydrate
MgSO 4 .7H 2 o
370
M-7774
Potassium dihydrogen phosphate anhydrous
K H 2 PO 4
170
P-8416
FeSO4.7H2O
27.8
F-8263
Ethylenediaminetetraacetic acid
(EDTA)
C 10 h 14 N 2 o 8 Na 2 .2H 2 o
(Na 2 EDTA)
37.3
E-6635
Micronutrients
KI
0.83
P-8166
h 3 BO 3
6.2
B-9645
MnSO 4 .H 2 o
6.9
M-7899
...
Embodiment 2
[0057] Example 2: Plant material
[0058] 1.5-month-old immature green pods were collected from 20-meter-tall mature Acacia mangium Willd trees along Science Park Avenue, Singapore. Gently separate the green seeds from the pods and rinse with sterile double deionized water (ddH 2 O) wash 4-5 times. The surface of the green seeds was disinfected with 30% commercially available sodium hypochlorite bleach (NaClO) for 5 min and washed with sterile ddHO 2 After washing 5 times with O, the seeds were washed with 0.1% HgCl 2 Sterilize for 6 minutes, and use sterile ddH 2 O wash 5 times. The seed coat is removed with sterile forceps, and the embryos are isolated. Hypocotyls (EA) (Fig. 1A) were gently detached with sterile forceps and used as explants for embryogenic callus induction.
Embodiment 3
[0059] Embodiment 3: culture condition
[0060] After autoclaving at 121°C for 25 minutes, all media were adjusted to pH 5.8. The plant growth regulators tested in the experiment were added to the medium after autoclaving, but other substances were added to the medium before autoclaving. All cultures were maintained at 28°C at a luminance of 23.41-26.01 μmol.s -1 m -2 Range of warm white fluorescent lamps with a photoperiod of 16 hours.
PUM
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