玉米丛矮病类菌原体的提纯

 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。

PURIFICATION OF MYCOPLASMA-LIKE ORGANISM (MLO) ASSOCIATED WITH MAIZE BUSHY STUNT DISEASE

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℃ 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 CaCl₂ 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.