乙酰乳酸合酶活性的简易测定方法建立  

SIMPLIFIED ASSAY OF ACETOLACTATE SYNTHASE ACTIVITIES

作　　者：陈以峰[1] 李宜慰[1] 汤日圣[1] 李永丰[1] 梅传生[1] 翟其楷 刘宁政 

机构地区：[1]江苏省农科院遗传生理所,江苏省农科院现代化所,昆山市农业局,江宁县农业局

出　　处：《江西农业大学学报》1996年第2期213-218,共6页Acta Agriculturae Universitatis Jiangxiensis

摘　　要：乙酰乳酸合酶（ＥＣ４，１，３，１８）（简称ＡＬＳ）是磺酰脲类等新型除草剂的靶标。该酶催化２个丙酮酸（或１个丙酮酸与１个α－丁酮酸）形成乙酰乳酸（或乙酰羟丁酸）。利用间接比色法测定该酶活性，即将产物乙酰乳酸脱梭形成３－羟基丁酮，再与肌酸及甲萘酚形成粉红色复合物，该复合物在５３０ｎｍ处有最大吸收。本文对影响测定该酶活性的各种因素作了比较，认为提取液中省去甘油，提取液及反应液中省去ＦＡＤ，提高反应液中丙酮酸钠浓度至１００ｍｍｏ１／Ｌ，降低显色剂浓度至０．０８３％（体积分数）肌酸／０．８３％（质量分数）甲萘酚能提高该测定方法的准确性与灵敏度，降低成本，由此建立了适用于测定罔草等杂草ＡＬＳ活性的技术。Acetolactate synthase(EC 4.1.3.18,ALS) is known as the target of some new classes ofherbicides such as sullonylureas,imidazolinones;This enzyme catalyzes the reactions:2 Pyruvate→Acetolactate+CO_2 or Pyruvate + a-Ketobutyrate→Aceto hydrobutyrate+ CO_2.ALS activities were detected bv indirect spectrophotometry of the produet acetolactate at 530 nm.This assav contains two conversions:the end product acetolactate to acetoin and formation ofacetoin,creatine and naphthol complex.In this article,a number of factors involved in influencing ALS activities in calli from young panicles of the weed Backnlana syzigczchnek are com pared and analysed.As a result it is suggested that omission of glycerol in extract mixture andof FAD in extract and reaction mixtures,increase of sodium pyruvate level in reaction mixtureupto 100 mmo1/L,and reduction of creatine and naphthol levels respectively to 0.083%and0.83%could raise accuracy and sensitivity of this assay and lower assay cost.Therefore,thismodified assav has been established suitable for detection of ALS activities in weeds such asBackmana syzigachnek which grow in wheat fields.

关 键 词：乙酰乳酸合酶 活性 测定 绿黄隆 

分 类 号：Q55[生物学—生物化学] S482.4[农业科学—农药学]