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The ACC oxidase (ACO) activity was measured as the ability of plant tissue to convert exogenous ACC to ethylene. The homogenate was prepared in an extraction buffer as described byMoya-Leon and John (1994), with modifications. One g tomato pericarp tissue was homogenized using a cold pestle and mortar with 3mL extraction buffer which contained 0.1MTris–HCl (pH7.5), 5%PVP, 10%glycerol, 5.0 mM dithiothreitol, 0.1 mM FeSO4 and 30 mM sodium ascorbate. The homogenate was centrifuged at 10,000×g for 20min at 4 ◦C.The determination of ACO activity was carried out in sealed tubes with a reaction mixture containing 1.8mL of 0.1M Tris–HCl buffer (pH 7.5, containing 10% glycerol, 30mM sodium ascorbate, 30mM NaHCO3, 0.1mM FeSO4 and 1.0mM ACC) and 0.2mL of enzyme extract. After being incubated at 30 ◦C for 20 min, 1mL of the gaseous portion was removed and assayed for ethylene as described above. The ACO activity was expressed as nmol g−1 FWh−1. Extracts for lipoxygenase (LOX) activity assays were prepared according to the method of Surrey (1964), with slight modifications. Two g tomato pericarp tissue was homogenized using a cold pestle and mortar in 10mL of 50mM phosphate buffer (pH 7.0) at 4 ◦C and centrifuged at 10,000×g for 15min. The supernatant liquid was used as the enzyme source. The reaction mixture contained 25 L of 100mM linoleic acid sodium, 2.775mL of 100mM acetic acid buffer (pH 5.5) and 0.2mL of supernatant enzyme extract at 30 ◦C. A unit of LOX was defined as a change of 1 absorbance unit at 234 nm in 1 min using a UVIKON XL Ultraviolet spectrophotometer (Secomam, France). These determinations were made from three replicates sampled from 5 fruit. |
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5楼2011-03-20 14:01:45
昔年残梦(金币+2, 翻译EPI+1): 2011-03-20 13:50:38
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The ACC oxidase (ACO) activity was measured as the ability of plant tissue to convert exogenous ACC to ethylene. The homogenate was prepared in an extraction buffer as described by Moya-Leon and John (1994), with modifications. ACC氧化酶（ACO）的活性测定可以用来衡量植物组织将外源的ACC转化为乙烯的能力。这种匀浆由Moya-Leon和约翰在1994年提取的一种缓冲液中制备、描述并加以修饰。 |
2楼2011-03-20 13:39:44
昔年残梦(金币+4): 2011-03-20 13:52:09
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One g tomato pericarp tissue was homogenized using a cold pestle and mortar with 3mL extraction buffer which contained 0.1MTris–HCl (pH7.5), 5%PVP, 10%glycerol, 5.0 mM dithiothreitol, 0.1 mM FeSO4 and 30 mM sodium ascorbate. The homogenate was centrifuged at 10,000×g for 20min at 4 ◦C. The determination of ACO activity was carried out in sealed tubes with a reaction mixture containing 1.8mL of 0.1M Tris–HCl buffer (pH 7.5, containing 10% glycerol, 30mM sodium ascorbate, 30mM NaHCO3, 0.1mM FeSO4 and 1.0mM ACC) and 0.2mL of enzyme extract. 1克番茄果皮组织匀浆的制备需要用冷的研钵和杵拌着3毫升的提取缓冲液进行研磨，缓冲液中包含有0.1摩Tris -盐酸（pH7.5），5％的PVP，10％甘油，5.0毫摩二硫苏糖醇，0.1毫摩硫酸亚铁和30毫摩抗坏血酸钠。将这个匀浆在4度下，10，000转离心20分钟克。 ACO活性的测定：将其放在一个装有混合物的密封管中，混合物中包含有1.8毫升的0.1摩的Tris - HCl缓冲液（pH 7.5 ，含10％甘油，30毫摩抗坏血酸钠，30毫摩碳酸氢钠，0.1毫摩硫酸亚铁和1.0毫摩ACC）和0.2毫升酶提取物。 |
3楼2011-03-20 13:50:44
昔年残梦(金币+10): 2011-03-20 14:03:54
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After being incubated at 30 ◦C for 20 min, 1mL of the gaseous portion was removed and assayed for ethylene as described above. The ACO activity was expressed as nmol g−1 FWh−1.Extracts for lipoxygenase (LOX) activity assays were prepared according to the method of Surrey (1964), with slight modifications. Two g tomato pericarp tissue was homogenized using a cold pestle and mortar in 10mL of 50mM phosphate buffer (pH 7.0) at 4 ◦C and centrifuged at 10,000×g for 15min. 30度温育20分钟，其中1毫升的气态部分被去除，就能够检测到如上所述的乙烯成分。ACO的活性表现为nmol g−1 FWh−1。根据萨里法（1964年）所提出的方法，脂氧合酶（LOX）活性检测的提取物被制备，但略有修改。两g番茄果皮组织匀浆使用上述同样方法，用10毫升50毫摩磷酸盐缓冲液（pH值7.0）在4◦ç，10000 ×转下离心15分钟。 |
4楼2011-03-20 13:58:15
