影响决明无菌苗子叶原生质体分离和培养因素的研究

以决明(Cassia obtusi folia)无菌苗子叶为材料,对酶组合、无菌苗日龄,植物激素组合和培养方法对其原生质体的分离和培养的影响进行了研究。结果表明:用3％的纤维素酶和0.2％Pectinase Y-23的酶组合处理决明无菌苗子叶块8小时可以高效分离出有活力的原生质体;约14日龄的决明无菌苗子叶比较适合于原生质体的分离;适当浓度的2,4-D 有利于原生质体的分离。促进原生质体分裂的理想的植物激素组合为0.4 mg/L 2,4-D,1.0 mg/L NAA and 0.1 mg/L KT;漂浮培养法最有利于原生质体的分裂和发育。找出了适合于决明无菌苗子叶原生质体的分离和培养的酶组合、植物激索组合、有效培养方法和决明无菌苗子叶日龄。这为有效地从决明无菌苗子叶原生质体再生植株奠定了基础。

Factors influencing protoplast culture of the seedlings of Cassia obtusifolia (Leguminosae)

In this work,the cotyledons of Cassia obtusifolia seedlings were used as materials. The effects of some factors such as different combinations of enzyme mixture, day-age of C. obtusi folia seedlings, culture methods and phytohormones in medium, on protoplast isolation and culture were investigated. The results demonstrated that protoplasts with high yield and quality were obtained by treating the cotyledon pieces with a mixture of 3% cellulase(Onozuka R-10)and 0. 2% Pectinase Y-23 for 8h. The cotyledons from 14 day-old seedlings were appropriate for isolating protoplasts. The floating culture method was more suitable for protoplast division and the division frequency was 9. 7%. The proper phytohormone combination was 0. 4 mg/L 2, 4-D, 1. 0 mg/L NAA and 0. 1 rng/L KT 2,4-D,in which the division frequency 21. 2% was gotten. The optimum combination of enzyme mixture,phytohormone combination,culture method and proper day-age of C. obtusi folia seedlings were determined. This laid the foundation of efficient plant regeneration from cotyledonary protoplasts of C. obtusifolia seedlings.