青霉素-溶菌酶法分离蓝藻原生质球

ISOLATION OF BLUE-GREEN ALGAL SPHEROPLASTS BY PENICILLIN-LYSOZYME METHOD

本试验使用磷酸二氢钾、柠檬酸钠、酒石酸铵、硫酸铵、硝酸铵、硝酸钠、KCl和NaCl等8种物质组成的复合渗透稳定剂,每种浓度0.15mo1,等量混合,并加入原生质膜稳定剂(0.1％CaCl_2)。材料经青霉素预处理6小时,然后用溶菌酶处理。溶菌酶浓度0.2％。处理温度28°—30℃,处理时间3小时。所形成的原生质球为透明和膨大的圆球体,对低渗和机械处理极为敏感。

Osmotic stabilizer used in present study consists of eight compounds, i.e. ammonium sul- fate, ammonium tertrate, KH_2PO_4, sodium citrate, NH_4NO_3, NaNO_3, KCl and NaCl. Each of them was dissolved in to 0.5 mol then eight solutions were mixed in equal volume. A menbrane stabilizer CaCl_2(0.1%) was added to the mixed solution. The pH of the mixed solution was about 6.2. Blue-green algal trichomes for the spheroplast isolation were first treated with penicil--lin. Penicillin dosage and treating time were different according to different materials. After penicillin procedure, the materials were then treated with lysozyme (0.2%). Temperature of the treatment was 28--30℃, Spheroplast production started in 15 minutes after treatment and finished in 3 hours. The spheroplasts remained a chain form mostly and became single sphe- roid through centrifugation and resuspension. The spheroplasts were typical spherical, more transparent and sensitive to hypotonic condition or mechanical treatment.