BioTechnica International Inc., 85, Bolton Street, 02140, Cambridge, MA, USA.
Plant Cell Rep. 1992 Nov;11(12):627-31. doi: 10.1007/BF00236387.
A gene from Pseudomonas putida coding for a dehalogenase capable of degrading 2,2 dichloropropionic acid (2,2DCPA), the active ingredient of the herbicide dalapon, has been isolated and characterised. In plant transformation experiments the gene was shown to confer resistance to 2,2DCPA at a tissue culture level where 2,2DCPA could be used to select for transformants. At the whole plant level, transformed plants showed resistance to 2,2DCPA at concentrations up to 5 times the recommended dose rate of dalapon when it was sprayed on their leaves. At lower concentrations, the herbicide caused a non-lethal yellowing of sensitive plants which clearly distinguished them from resistant plants. The mode of action of chlorinated aliphatic acids is not known but they probably affect many enzyme pathways. The results described here are the first example of engineering a plant resistant to a herbicide that does not have one specific enzyme as its target site. This gene has several advantages as a marker in plant breeding and genetic studies. For example, the herbicide is readily available and has low toxicity, transformants can be selected at both the tissue culture and the whole plant level, a large number of transformed plants can easily be screened even in the field, and there is a very low probability of selecting spontaneous mutants.
从恶臭假单胞菌中分离出一种编码脱卤酶的基因,该脱卤酶能够降解除草剂达草灭的有效成分 2,2-二氯丙酸(2,2DCPA)。对该基因进行了鉴定和特性分析。在植物转化实验中,该基因在组织培养水平上赋予了对 2,2DCPA 的抗性,在组织培养水平上可以使用 2,2DCPA 来选择转化体。在整个植株水平上,当将达草灭喷洒在叶片上时,转化植株对浓度高达推荐剂量 5 倍的 2,2DCPA 表现出抗性。在较低浓度下,除草剂会导致敏感植物出现非致死性黄化,这使它们与抗性植物明显区分开来。氯化脂肪族酸的作用方式尚不清楚,但它们可能会影响许多酶途径。这里描述的结果是首次对植物进行工程改造,使其对不以特定酶为靶标的除草剂具有抗性。该基因在植物育种和遗传研究中有几个作为标记的优点。例如,该除草剂易于获得且毒性低,转化体可以在组织培养和整个植株水平上进行选择,可以很容易地对大量转化植物进行筛选,即使在田间也是如此,而且选择自发突变体的可能性非常低。