摘要
油菜叶片经油菜菌核病菌 （ Ｓｃｌｅｒｏｔｉｎｉａ ｓｃｌｅｒｏｔｉｏｒｕｍ ） 或水溶性壳聚糖处理后， 几丁质酶活性明显提高， 分别在处理后 １ ｄ 和 ２ ｄ 达活性高峰， 比对照分别高５ 倍和２ 倍左右。被激发提高的几丁质酶表现为内切酶活性， 细胞间的几丁质酶比活力显著高于细胞内的酶比活力。菌核病菌侵染后的油菜叶片提取液经６０％ 饱和度的硫酸铵沉淀，再通过再生几丁质柱亲和层析得到了纯化的几丁质酶， 经 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 显示单一的谱带， 其相对分子质量为 ３２ ０００。纯化的油菜叶片几丁质酶能降解油菜菌核病菌菌丝细胞壁， 在马铃薯葡萄糖琼脂培养基上可抑制菌丝生长。
Rape( Brassica napus )leaves were treated by water soluble chitosan and/or Sclerotinia sclerotiorum, a pathogen causing root rot of rape.The chitinase in the leaves increased rapidly.In pathogen infected leaves,chitinase activity started to increase after a lag 12 h and was about 6 times as high as that in the control within 24 h.While in the chitosan treated leaves,chitinase activity started to increase after treatment for 24 h,and was about 3 times as high as that in the control within 48 h.The chitinase in rape leaves showed endochitinase activity and had higher specific activity in extract of intercellular fluid extracted by vacuum infiltration than that in leaf cells.Chitinase in rape leaves treated by S.sclerotiorum was purified by precipitate with ammonium sulfate at a saturation of 60% followed by affinity chromatography on a column of regenerated chitin.The purified enzyme showed a single protein band on sodium dodecyl sulfate polyacrylamide gel electrophoresis with molecular weight about 32 000.The enzyme inhibited the mycelium radial growth of S.sclerotiorum on potato dextrose agar medium and lysised the mycelium cell wall.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
1999年第3期41-44,共4页
Journal of Nanjing Agricultural University
基金
江苏省 "九五"农业科技重点攻关项目
关键词
油菜
几丁质酶
壳聚糖
菌核病菌
纯化
rape
chitinase
chitosan
Sclerotinia sclerotiorum
purification
