TH11202Y - A culture medium formula to induce callus formation in tissue culture from bougainvillea leaves. - Google Patents

Abstract

DC60 (August 25, 2015) A study of callus-inducing culture media formulas in tissue culture using bougainvillea leaves found that the best formula was MS medium. The chemicals used for preparing the MS culture medium in the specified amounts followed the standard formula. Amount in concentrated solution (mg/L) 100 times (g/L) Main nutrient solution MS CaCl12.2H2O (Calcium chloride) 440.00 44 KNO3 (Potassium nitrate) 1,990.00 190 KH2PO4 (Potassium dihydrophosphate) 170.00 17 MgSO4.7H2O (Magnesium sulfate) 370.00 37 NH4NO3 (Ammonium nitrate) 1,650.00 165.00 Secondary nutrient solution MS MnSO4.4H2O (Manganese sulfate) 16.90 1.69 ZnSO4.7H2O (Zinc sulfate) 8.60 0.86 H3BO3 (Boric acid) 6.20 0.62 KI (Potassium iodide) 0.83 0.083 Na2MoO4.2H2O (Sodium molybdate) 0.25 0.025 CoCl2.6H2O (Cobalt chloride) 0.025 0.0025 CuSO4.5H2O (Copper sulfate) 0.025 0.0025 Concentrated iron solution (MS): FeSO4.7H2O (Ferrous sulfate) 27.30 2.73 Na2EDTA (Sodium EDTA) 37.30 3.73 Concentrated organic solution (MS): Myo-inositol 100.00 10.00 Glycine 2.00 0.20 Thiamine.HCl (Thiamine hydrochloride) 0.10 0.01 Pyridoxin HCl (pyridoxine hydrochloride) 0.50 0.05 Nicotinic acid 0.50 0.05 Add 30.00 g/L of sugar and 8.00 g of agar and stir until the agar dissolves. Add the growth regulator cytokinin, BA (N6-benzyladenine), at a concentration of 1.00 mg/L, along with the hormone auxin, NAA (alpha-naphthalene acetic acid), at a concentration of 0.10 mg/L. The MS medium was the most effective in inducing callus formation, with an average callus size of 1.15 cm. All calluses were loosely clustered (friable callus) and were light green in color. Callus formation occurred within 25 days of culture. (Revised 7/09/2015 (25/08/2015)) A study of callus-inducing culture media formulas using bougainvillea leaves revealed that the best formula was MS medium. The chemicals used in preparing the MS medium were specified in standard quantities. Amount in concentrated solution (mg/L) 100 times (g/L) Main nutrient solution MS CaCl12.2H2O (Calcium chloride) 440.00 44.00 KNO3 (Potassium nitrate) 1,900.00 190.00 KH2PO4 (Potassium dihydrophosphate) 170.00 17.00 MgSO4.7H2O (Magnesium sulfate) 370.00 37.00 NH4NO3 (Ammonium nitrate) 1,650.00 165.00 Secondary nutrient solution MS MnSO4.4H2O (Manganese sulfate) 16.90 1.69 ZnSO4.7H2O (Zinc sulfate) 8.60 0.86 H3BO3 (Beric acid) 6.20 0.62 KI (Potassium Iodide) 0.83 0.083 Na2MoO4.2H2O (Sodium Molybdate) 0.25 0.025 CoCl2.6H2O (Cobalt Chloride) 0.025 0.0025 CuSO4.5H2O (Copper Sulfate) 0.025 0.0025 Concentrated Iron Solution (MS) FeSO4.7H2O (Ferrous Sulfate) 27.30 2.73 Na2EDTA (Sodium EDTA) 37.30 3.73 Concentrated Organic Solution (MS) Myo-inositol 100.00 10.00 Glycine 2.00 0.20 Thiamine.HCl (Thiamine Hydrochloride) 0.10 0.01 Pyridoxin HCl (pyridoxine hydrochloride) 0.50 0.05 Nicotinic acid 0.50 0.05 Add 30.00 g/L of sugar and 8.00 g of agar, then boil until the agar dissolves. Add the growth regulator cytokinin, BA (N6-benzyladenine), at a concentration of 1.00 mg/L, along with the hormone auxin, NAA (Naphthalene acetic acid), at a concentration of 0.10 mg/L. This induced the best callus formation, with an average callus size of 1.15 cm. All calluses were loosely attached (friable callus) and were light green in color. Callus formation occurred within 25 days after culture. --------------------------------------------------------------------------------------- A study of callus-inducing culture media formulas from Bougainvillea hybrida tissue culture revealed that the best formula was MS medium supplemented with the cytokinin growth regulator BA (N6-benzyladenine) at a concentration of 1.00 mg/L, along with the auxin hormone NAA (Naphthalene acetic acid) at a concentration of 0.10 mg/L. This formula induced the best callus formation, with an average callus size of 1.15 cm. All calluses were loosely clustered (friable callus) and were light green in color. Callus formation occurred within 25 days after culture. (Revised August 7, 2014) Further studies on callus-inducing culture media formulas using Bougainvillea hybrida leaves showed that the best formula was MS medium. The chemicals used for preparing the MS culture medium in standard quantities were... Amount in concentrated solution (mg/L) 100 times (g/L) Main nutrient solution MS CaCl12.2H2O (Calcium chloride) 440.00 44 KNO3 (Potassium nitrate) 1990.00 190 KH2PO4 (Potassium dihydrophosphate) 170.00 17 MgSO4.7H2O (Magnesium sulfate) 370.00 37 NH4NO3 (Ammonium nitrate) 1,650.00 165 Secondary nutrient solution MS MnSO4.4H2O (Manganese sulfate) 16.90 1.69 ZnSO4.7H2O (Zinc sulfate) 8.60 0.86 H3BO3 (Boric acid) 6.20 0.62 KI (Potassium iodide) 0.83 0.083 Na2MoO4.2H2O (Sodium molybdate) 0.25 0.025 CoCl2.6H2O (Cobalt chloride) 0.025 0.0025 CuSO4.5H2O (Copper sulfate) 0.025 0.0025 Concentrated iron solution (MS): FeSO4.7H2O (Ferrous sulfate) 27.30 2.73 Na2DETA (Sodium EDTA) 37.30 3.73 Concentrated organic solution (MS): Myo-inositol 100.00 10 Glycine 2.00 0.2 Thiamine.HCl (Thiamine hydrochloride) 0.10 0.01 Pyridoxin.HCl Pyridoxine hydrochloride 0.50 0.05 Nicotinic acid 0.50 0.05, sugar 30.00 g/L and agar 8.00 g were added and the mixture was stirred until the agar dissolved. A growth regulator from the cytokinin group, BA (N6-benzyladenine), at a concentration of 1.00 mg/L was added, along with the hormone auxin, NAA (Naphthalene acetic acid), at a concentration of 0.10 mg/L. This induced callus formation most effectively. The callus had an average size of 1.15 cm, were all loosely attached (friable callus), and were light green in color. Callus formation occurred within 25 days of cultivation.

Claims (1)

Revised 7/09/2015 (25/08/2015) 1. The culture medium formula for callus induction in bougainvillea tissue culture consists of: a. 100x concentration MS primary nutrient solution, 10.00 ml. b. 100x concentration MS secondary nutrient solution, 10.00 ml. c. 100x concentration MS iron solution, 10.00 ml. d. 100x concentration organic matter solution, 5.00 ml. e. BA (N6-benzyl adenine) growth regulator, 1.00 ml. f. NAA (naphthalene acetic acid) growth regulator, 0.10 ml. g. Sugar, 30.00 ml. h. Distilled water, adjusted to 1000 ml. i. Agar, 8.00 ml. 2. A culture medium formula that induces callus formation in bougainvillea leaf tissue culture. According to claim 1, the MS primary nutrient solution, 100x concentration, contains: - CaCl12.2H2O (calcium chloride) 44 g - KNO3 (potassium nitrate) 190 g - KH2PO4 (potassium dihydrophosphate) 17 g - MgSO4.7H2O (magnesium sulfate) 37 g - NH4NO3 (ammonium nitrate) 165 g - distilled water, final volume adjusted to 1000 ml. The MS secondary nutrient solution, 100x concentration, contains: - MnSO4.4H2O (manganese sulfate) 1.69 g - ZnSO4.7H2O (zinc sulfate) 0.86 g - H3BO3 (beric acid) 0.62 g - KI (potassium iodide) 0.083 g - A concentrated MS (Multiplier) iron solution (100x concentration) contains: - FeSO4.7H2O (Ferrous Sulfate) 2.73 g - Na2EDTA (Sodium Hydrochloride) 0.0025 g - CuSO4.5H2O (Copper Sulfate) 0.0025 g - Distilled water. Adjust the final volume to 1000 ml. (Pyridoxine hydrochloride) 0.05 g - Nicotinic acid 0.05 g - Distilled water. Adjust the final volume to 1,000 ml. ------------------------------------------------------ 1. Tissue culture medium formula for Bougainvillea hybrida leaves to induce callus formation. Consists of MS medium supplemented with the growth regulator BA (N6-benzyl adenine) at a concentration of 1.00 mg/L combined with NAA (Napthalene acetic acid) 0.10 mg/L. (Revised August 8, 2014) 1. The callus-inducing culture medium formula for tissue culture using bougainvillea leaves consists of: a. 100x concentration MS primary nutrient solution, 10.00 ml. b. 100x concentration MS secondary nutrient solution, 10.00 ml. c. 100x concentration MS iron solution, 10.00 ml. d. 100x concentration organic solution, 5.00 ml. e. BA (N6-benzyl adenine) growth regulator, 1.00, 3.00, or 5.00 ml. f. NAA (naphthalene acetic acid) growth regulator, 0.10 ml. g. Sugar, 30.00 g. h. Distilled water, adjusted to 1000 ml. i. Agar, 8.0 g.