摘要
本文报道一种提纯玉米丛矮病类菌原体的方法:将病玉米组织在捣碎机中低速去除薄壁组织;用纤维素酶、果胶酶和半纤维酶处理,35℃下1小时;高速离心沉淀;进行二次Percoll密度梯度离心。间接酶联免疫吸附试验和电镜检查结果表明,利用此方法能够得到高纯度、完繁的类菌原体。
An improved method was developed for the purification of mycoplasmalike organism associated with maize bushy stunt disease. Diseased corn plants were crushed with a tissue homogennizer using slow and gentle cycle until most of the parenchyma tissue was broken.The retained vascular tissues were then incubated at 35鈩?for 1 hr in an enzyme solution containing 0.8% (W/V) cellulose, 0.2% (W/V) pectinase, 0.2% (W/V) hemicellulose, 1 mM CaCl_2 and 0.3 M mannitol. The suffered tissues were macerated with a tissue grinder and the suspension solution was subjected to one cycle of low and high speed centrifugation. The pellets were suspended and centrifugated twice in discontinuous Percoll density gradient.Indirect enzyme-linked immunosorbent assay and electron microscopy confirmed that highly purified and intact MLO could be obtained with this method.
出处
《植物病理学报》
CAS
CSCD
北大核心
1991年第1期27-30,共4页
Acta Phytopathologica Sinica
