罗布麻愈伤组织原生质体分离培养  

Preliminary Study on the Isolation and Culture of Protoplast from Apocynum venetum Callus

作　　者：徐琴[1] 王晓军[1] 郝秀英[2] 刘敏[1] 康喜亮[1] 波拉提[1] 

机构地区：[1]中国科学院新疆理化技术研究所,新疆乌鲁木齐830011 [2]新疆农业科学院微生物应用研究所,新疆乌鲁木齐830091

出　　处：《安徽农业科学》2010年第3期1139-1140,共2页Journal of Anhui Agricultural Sciences

摘　　要：[目的]探究罗布麻愈伤组织原生质体的分离培养及其影响因素。[方法]以罗布麻无菌苗子叶与下胚轴诱导的愈伤组织为材料,研究MS培养基附加不同激素组合对原生质体分离培养的影响以及愈伤组织继代培养、不同浓度纤维素酶与离析酶组合、DPD和MS等4种培养基对原生质体产量的影响。[结果]激素组合中以6-BA 0.5 mg/L+NAA 0.5 mg/L组合获得最高愈伤组织诱导率,达100%;用固体培养基进行继代培养的愈伤组织的原生质体产量高于液体培养基,尤其以从继代1次愈伤组织收集的原生质体最为适宜;纤维素酶0.30%+离析酶0.30%组合获得的原生质体产量最高,且细胞碎片较少;DPD培养基的原生质体较其他培养基的分裂得早,MS培养基的培养效果最差。[结论]该研究为罗布麻遗传转化、突变体筛选及体细胞融合育种提供了技术途径。[ Objective] The study aimed to explore the protoplast isolation and cuhure of callus in Apocynum venetu.m and the influence factors. [ Method] With callus induced from the cotyledon and hypoeotyts in aseptic seedlings of A. venetum as material, the effeets of MS medium supplemented with different hormone combinations on the protoplast isolation and culture and the effects of the eallus subeulture, different eonen, of cellulase enzymes combined with the maceration enzyme, 4 kinds of culture mediums sueh as DPD and MS on the protoplast yield were researched. [ Result] Hormone combination with 6-BA 0.5 mg/L ＋ NAA 0.5 mg/L combination received the highest eallus induction rate, up to 100% ; The protoplast yield of the callus cuhured on solid medium for subculture was higher than that of the callus cultured on the liquid medium and specially the protoplasts collected from the callus subeuhured for once were most suitable. The protoplast yield obtained with the combination of cellulase enzyme 0.30% ＋ maceration enzyme 0.30% was the highest, with the less segments of cells. The division of the protoplast cuhured on DPD medium was earlier than that on other mediums and the cuhure effect of MS medium was the worst. [ Conclusion] This study provided a technical approach for the genetic transformation, mutant screening and somatic fusion breeding of A. venetum.

关 键 词：罗布麻 愈伤组织 原生质体 

分 类 号：S563.7[农业科学—作物学]