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水稻组织培养－分化生根的激素
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生长素auxin
植物内源生长素：吲哚乙酸（IAA），吲哚丁酸（IBA），苯乙酸（PAA）
结构特点：有一个芳香环和羧基侧链，二者之间有一个芳香环或氧原子间隔。
人工合成生长素：2，4-二氯苯氧乙酸（2，4- D）和α-萘乙酸NAA
一种人工合成的具有与生长素类似生理效应的有机物。有效成分为α-NAA，分子式C12H10O2，白色结晶，不溶于水，溶于酒精等有机溶剂，其钠盐溶于水，效应与NAA相同，但应用浓度较高。生产容易，活性强，为农业及园艺上使用最广泛的人工合成生长素之一。它可以促进发芽、单性结实及花芽形成（波罗），抑制器官脱落以及刺激插枝生根等。由于生根量少且较粗，故宜与吲哚丁酸公用（刺激插枝生根数目多且较细）。
是广谱型植物生长调节剂，能促进细胞分裂与扩大，诱导形成不定根增加坐果，防止落果，改变雌、雄花比率等。可经叶片、树枝的嫩表皮，种子进入到植株内，随营养流输导到全株。
水稻生根培养基中加入生长素吲哚丁酸IBA 0.5 mg/L 促进生根。
细胞分裂素：在生长素存在的条件下诱导愈伤组织细胞分裂，适当比例下促经愈伤不定芽的发生。培养基中缺乏KT烟草愈伤细胞分裂受抑制，
较高的生长素和激动素的比列2：0.02 mg/L促进根的分化，
中等的生长素和激动素的比列为2：0.2 mg/L维持愈伤的生长，
较低的生长素和激动素的比列为2：0.5 mg/L促进芽的分化。
当培养基中[CTK]/[IAA]的比值高时,诱导愈伤组织形成芽;当[CTK]/[IAA]的比值低时,诱导愈伤组织形成根;
人工合成的细胞分裂素
激动素(KT，kinetin) 一种腺嘌呤衍生物－6-呋喃氨基腺嘌呤
6-苄基腺嘌呤(6-benzyl adenine,BA，6-BA)和四氢吡喃苄基腺嘌呤(tetrahydropyranyl benzyladenine,又称多氯苯甲酸，简称PBA)等。
The importance of plant growth regulators in plant tissue culture is well documented. Sigma offers a broad range of plant growth regulators specifically tested for plant cell culture. Each product is assayed for physical and chemical characteristics, then is biologically tested following the criteria established for powdered media. Each auxin is tested for enhancement of callus growth and/or root initiation in vitro. Each cytokinin is tested for stimulation of shoot production.
The plant growth regulator solutions are sterile filtered through a double 0.2 µm filter unit into sterile bottles. Each lot of plant growth regulator solution is tested for sterility according to specifications established by U.S. Pharmacopeia Vol. XXI, as well as biological testing in plant cell culture using criteria identical to those established for our plant tissue culture media.
The plant growth regulator solutions are sterile filtered through a double 0.2 µm filter unit into sterile bottles. Each lot of plant growth regulator solution is tested for sterility according to specifications established by U.S. Pharmacopeia Vol. XXI, as well as biological testing in plant cell culture using criteria identical to those established for our plant tissue culture media.
Preparation and Use
To prepare a 1 mg/ml stock solution: Add 100 mg of the plant growth regulator to a 100 ml volumetric flask or other glass container. Add 2-5 ml of solvent to dissolve the powder. Once completely dissolved, bring to volume with double processed water (Product No. W 3500). Stirring the solution while adding water may be required to keep the material in solution. Store the stock solution as recommended in the tables. Add 1.0 ml of the stock solution to 1 liter of medium to obtain a final concentration of 1.0 mg/L of the plant growth regulator in the culture medium. (See Table below.)
Volume of Stock Solution = (Desired Hormone Conc. X Medium Volume) / Stock Solution Conc.
Plant Cell Culture Tested Auxins are generally used in plant cell culture at a concentration range of 0.01-10.0 mg/L. When added in appropriate concentrations, they may regulate cell elongation, tissue swelling, cell division, formation of adventitious roots, inhibition of adventitious and axillary shoot formation, callus initiation and growth, and induction of embryogenesis.
Plant Cell Culture Tested Cytokinins are generally used in plant cell culture at a concentration range of 0.1-10.0 mg/L. When added in appropriate concentrations, they may regulate cell division, stimulate axillary and adventitious shoot proliferation, regulate differentiation, inhibit root formation, activate RNA synthesis and stimulate protein and enzyme activity.
Plant Cell Culture Tested Cytokinins are generally used in plant cell culture at a concentration range of 0.1-10.0 mg/L. When added in appropriate concentrations, they may regulate cell division, stimulate axillary and adventitious shoot proliferation, regulate differentiation, inhibit root formation, activate RNA synthesis and stimulate protein and enzyme activity.
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Product
Name |
Product
Number |
Molar Equivalence
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Solution Preparation
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Mol. Wt.
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µM for 1mg/L
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Solvent
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Diluent
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Powder Storage
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Liquid Storage
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Steriliz-
ation* |
Working Conc. (mg/L)
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p-Chlorophenoxyacetic acid (4-CPA)4-氯苯氧乙酸
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186.6
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5.36
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EtOH
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—
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RT
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2-8°C
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CA
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0.1-10.0
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2,4-Dichlorophenoxyacetic acid 2,4-二氯苯氧基乙酸
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221
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4.53
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—
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—
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RT
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2-8°C
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CA
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0.01-6.0
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2,4-Dichlorophenoxyacetic acid Sodium salt
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243
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4.12
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Water
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—
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RT
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2-8°C
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CA
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0.01-6.0
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Indole-3-acetic acid Free acid (IAA)
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175.2
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5.71
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EtOH/1N NaOH
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Water
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-0°C
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-0°C
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CA/F
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0.01-3.0
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Indole-3-acetic acid Sodium salt
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197.2
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5.07
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Water
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Water
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2-8°C
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-0°C
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CA/F
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0.01-3.0
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Indole-3-acetic acid methyl ester
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189.2
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5.29
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—
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—
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2-8°C
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2-8°C
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—
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—
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Indole-3-acetyl-L-aspartic acid
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290.3
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3.45
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0.5N NaOH
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Water
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-0°C
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-0°C
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F
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0.01-5.0
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Indole-3-butyric acid (IBA)
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203.2
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4.90
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EtOH/1N NaOH
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Water
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2-8°C
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-0°C
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CA/F
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0.1-10.0
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Indole-3-butyric acid Potassium salt (K-IBA)
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241.3
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4.14
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Water
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—
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2-8°C
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-0°C
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CA/F
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0.1-10.0
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alpha-Naphthaleneacetic acid Free acid (NAA)
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186.2
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5.37
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1N NaOH
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Water
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RT
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2-8°C
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CA
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0.1-10.0
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beta-Naphthoxyacetic acid Free acid (NOA)
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202.2
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4.95
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1N NaOH
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Water
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RT
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2-8°C
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CA
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0.1-10.0
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Phenylacetic acid (PAA)
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136.2
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7.34
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EtOH
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—
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RT
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2-8°C
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CA/F
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0.1-50.0
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Picloram
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241.5
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4.14
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DMSO
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—
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RT
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2-8°C
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CA
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0.01-10.0
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2,4,5-Trichlorophenoxyacetic acid (2,4,5-T)
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255.5
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3.91
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EtOH
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—
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RT
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2-8°C
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CA
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0.01-5.0
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2,3,5-Triiodobenzoic acid Free acid (TIBA)
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499.8
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2.00
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1N NaOH
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Water
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-0°C
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-0°C
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F
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0.05-5.0
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*CA = coautoclavable with other media components. F = filter sterlize. CA/F = coautoclavable with other media components, however, some loss of activity may occur. This can be compensated for by increasing component concentration. Component may be filter sterilized.
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Product
Name |
Product
Number |
Molar Equivalence
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Solution Preparation
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Mol. Wt.
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µM for 1mg/L
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Solvent
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Diluent
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Powder Storage
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Liquid Storage
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Steriliz-
ation* |
Working Conc. (mg/L)
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Adenine Free base
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135.1
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7.40
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1.0 HCl
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Water
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RT
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2-8°C
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CA
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50-250
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Adenine hemisulfate Hemisulfate salt
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184.2
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5.43
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Water
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—
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RT
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2-8°C
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CA
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50-250
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6-Benzylaminopurine (BA) 6-苄氨基嘌呤
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225.3
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4.44
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1N NaOH
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Water
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RT
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2-8°C
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CA/F
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0.1-5.0
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6-Benzylaminopurine Hydrochloride
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261.7
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3.82
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Water
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—
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RT
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2-8°C
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CA/F
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0.1-5.0
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6-Benzylaminopurine (BA)
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225.3
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4.44
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1N NaOH
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Water
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RT
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2-8°C
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CA/F
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0.1-5.0
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N-Benzyl-9-(2-tetrahydropyranyl)adenine (BPA)
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309.4
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3.23
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EtOH
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—
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-0°C
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-0°C
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CA/F
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0.1-5.0
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N-(2-Chloro-4-pyridyl)-N'-phenylurea (4-CPPU)
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247.7
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4.04
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DMSO
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—
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2-8°C
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2-8°C
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F
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0.001-1.0
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6-(gamma,gamma-Dimethylallylamino)purine (2iP)
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203.2
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4.92
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1N NaOH
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Water
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-0°C
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-0°C
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CA/F
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1.0-30.0
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6-(gamma,gamma-Dimethylallylamino)purine (2iP)
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203.2
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4.92
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1N NaOH
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Water
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-0°C
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-0°C
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CA/F
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1.0-30.0
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1,3-Diphenylurea (DPU)
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212.3
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4.71
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DMSO
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—
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RT
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2-8°C
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F
|
0.1-1.0
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Kinetin
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215.2
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4.65
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1N NaOH
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Water
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-0°C
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-0°C
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CA/F
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0.1-5.0
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Kinetin
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215.2
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4.65
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1N NaOH
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Water
|
-0°C
|
-0°C
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CA/F
|
0.1-5.0
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Kinetin
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215.2
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4.65
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1N NaOH
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Water
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-0°C
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-0°C
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CA/F
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0.1-5.0
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Kinetin Hydrochloride
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251.7
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3.97
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Water
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—
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-0°C
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-0°C
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CA/F
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0.1-5.0
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1-Phenyl-3-(1,2,3-thiadiazol-5-yl)urea
|
220.2
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4.54
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DMSO
|
—
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RT
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2-8°C
|
CA/F
|
0.001-0.05
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trans-Zeatin Free base
|
219.2
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4.56
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1N NaOH
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Water
|
-0°C
|
-0°C
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CA/F
|
0.01-5.0
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Zeatin
|
219.2
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4.56
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1N NaOH
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Water
|
-0°C
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-0°C
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CA/F
|
0.01-5.0
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trans-Zeatin Hydrochloride
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255.7
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3.91
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Water
|
—
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-0°C
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-0°C
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CA/F
|
0.01-5.0
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trans-Zeatin riboside
|
351.4
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2.85
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1N NaOH
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Water
|
-0°C
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-0°C
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F
|
0.01-5.0
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*CA = coautoclavable with other media components. F = filter sterlize. CA/F = coautoclavable with other media components, however, some loss of activity may occur. This can be compensated for by increasing component concentration. Component may be filter sterilized.
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Product
Name |
Product
Number |
Molar Equivalence
|
Solution Preparation
|
||||||
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Mol. Wt.
|
µM for 1mg/L
|
Solvent
|
Diluent
|
Powder Storage
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Liquid Storage
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Steriliz-
ation* |
Working Conc. (mg/L)
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(±)-cis,trans-Abscisic acid (ABA)
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264.3
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3.78
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1N NaOH
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Water
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-0°C
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-0°C
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CA/F
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0.1-10.0
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Ancymidol
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256.3
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3.90
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DMSO
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—
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2-8°C
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-0°C
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CA/F
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1.0-10.0
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Chlorocholine chloride (CCC)
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158.1
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6.33
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Water
|
—
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RT
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2-8°C
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F
|
up to 500
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3,6-Dichloro-o-anisic acid (Dicamba)
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221.0
|
4.52
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EtOH/Water
|
—
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2-8°C
|
2-8°C
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F
|
0.01-10.0
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Gibberellic acid (GA3)
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346.4
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2.89
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EtOH
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—
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RT
|
2-8°C
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CA/F
|
0.01-5.0
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Gibberellic acid Potassium salt (K-GA3)
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384.5
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2.60
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Water
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—
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2-8°C
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-0°C
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CA/F
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0.01-5.0
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Gibberellin A4 Free acid (GA4)
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332.4
|
3.01
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EtOH
|
—
|
-0°C
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-0°C
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F
|
0.01-5.0
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(±)-Jasmonic acid
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210.3
|
4.76
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EtOH
|
—
|
2-8°C
|
-0°C
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F
|
0.01-100.0
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Phloroglucinol
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126.1
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7.93
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Water
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—
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RT
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2-8°C
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CA/F
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up to 162
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N-(Phosphonomethyl)glycine (Glyphosate)
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169.1
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5.91
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1N NaOH
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Water
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RT
|
2-8°C
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F
|
—
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Succinic acid 2,2-dimethylhydrazide
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160.2
|
6.24
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Water
|
—
|
2-8°C
|
2-8°C
|
CA/F
|
0.1-10.0
|
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|
*CA = coautoclavable with other media components. F = filter sterlize. CA/F = coautoclavable with other media components, however, some loss of activity may occur. This can be compensated for by increasing component concentration. Component may be filter sterilized.
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