鹰嘴紫云英甲硫氨酸抗性系原生质体培养及植株再生

本研究建立了鹰嘴紫云英(AstragaluscicerL.)甲硫氨酸抗性系原生质体再生植株的实验体系。以茎切段诱导的松软愈伤组织为材料,通过酶法游离出大量有活力的原生质体。原生质体经培养持续细胞分裂形成了愈伤组织,并分化出再生苗。比较了不同培养基、培养密度对原生质体形成细胞分裂和再生的影响。结果表明,原生质体以2×105个/ml的植板密度,在附加2.0mg/L2,4-二氯苯氧乙酸(2,4-D)、0.2mg/L6-苄氨基嘌呤(6-BA)、200mg/L水解酪蛋白、2%蔗糖和0.3mol/L甘露醇DPD培养基中培养后,其分裂频率达38.3%。原生质体培养形成的愈伤组织仍具有对甲硫氨酸的抗性。转移到附加10mg/LKT、0.5mg/LNAA的MS分化培养基上,获得大量的再生苗。

PROTOPLAST CULTURE AND PLANT REGENERATION OF THE METHIONINE RESISTANT VARIANT OF ASTRAGALUS CICER L.

A protoplast-to-plant system for the methionine resistant variant of Astragalus cicer L. has been developed. The friable calli induced from stem segments of variant plants were used as materials for protoplast isolation through enzyme digestion. The effects of different media and plating densities on protoplast divisions and plant regeneration were studied. Sustained cell divisions and colony formation from the protoplasts of the methionine resistant cell line of Astragalus cicer L. were obtained by a DPD medium containing 2.0 mg/L 2,4- dichlorophenoxyacetic acid (2,4-D), 0.2 mg/L 6 -benzylaminopurine(6-BA), 0.3 mol/L mannitol, 200 mg/L casein hydrolysate and 2% (W/V) sucrose at a plating density of 2x10(5) /ml. The division frequency was 38.3%. At the same time, different dividing types of protoplasts were found. Organogenesis and shoot formation from the protoplast-derived calli were induced on MS medium supplemented with 0.5 mg/L NAA, 10 mg/L KT and 2% (W/V) sucrose. The protoplast-derived calli still expressed resistance to methionine. The protoplast to plant regeneration protocol developed in this study might provide the foundation for the resistant cell line as a parent for somatic hybridization.