我国主要栽培木薯品种体细胞胚胎发生与芽器官发生的研究  

A Study on Somatic Embryogenesis and Shoot Organogenesis of Major Cassava Cultivars in China

作　　者：赵姗姗[1] 李海霞[1] 刘佳 黄晨 马秋香 张鹏[1,2] 

机构地区：[1]中国科学院上海生命科学研究院植物生理生态研究所合成生物学重点实验室,上海200032 [2]植物分子遗传国家重点实验室,上海200032

出　　处：《农业生物技术学报》2010年第1期37-44,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(30771366);中国科学院知识创新工程重要方向项目(KSCX2-YW-G-026;KSCX2-YW-G-035);973计划(2010CB126605);上海市浦江人才计划(08PJ1410900);国家木薯产业技术体系项目(nycytx-17)共同资助

摘　　要：本研究对我国10个木薯(Manihot esculenta Crantz)主要栽培品种离体培养,研究结果表明不同品种之间在体细胞胚胎发生和器官发生的能力上因基因型不同而差异显著。以组培苗茎段侧芽或顶芽为外植体,在添加Picloram和2,4-D的体细胞胚胎发生诱导培养基上暗培养2周后,可诱导胚状体的形成。生长素浓度中,2,4-D在6mg/L及Picloram在12mg/L为最佳使用浓度,并且Picloram的效果明显优于2,4-D。NZ188、SC124和SC205体细胞胚胎发生的频率可达到80%以上。诱导的胚状体经循环继代培养可形成次生胚状体。次生胚状体在成熟培养基上经2～3周的光培养可形成绿色子叶的体细胞胚,进而萌发形成再生植株。NZ188及SC8体细胞胚萌发频率最好,可达80%以上。以培养2周的子叶胚切块置于体细胞胚胎发生诱导培养基上可诱导形成次生胚状体,诱导频率都在77%以上。子叶胚切块置于器官发生培养基上可诱导芽器官发生,2周后可形成芽原基及不定芽,诱导频率在34.4%到72.2%之间,其中NZ188、NZ199和SC124诱导效果较好。再生培养附加乙烯抑制剂AgNO3可明显抑制愈伤组织的形成,提高芽器官发生的能力,包括再生频率及芽再生的数目,浓度以4mg/L为宜。从含AgNO3的培养基上再生的芽有利于进一步生长和移栽成活。Study on in vitro cultures of ten cassava（Manihot esculenta Crantz） cultivars cultivated mainly in Chi na demonstrated the significant different capacities of somatic embryogenesis（SE） and shoot organogenesis（SO） among tested genotypes in this research.Using apical and axillary buds as explants,SE was induced on induction medium containing 12 mg/L picloram under the dark condition.After 2 weeks culturing,their SE frequencies were recorded.Among tested cultivars,NZ188,SC124 and SC205 could be over 80% on their SE frequencies.For SE induction,the effect of 12 mg/L picloram was better than that of 6 mg/L 2,4-D.Secondary SE could be established during cyclic subculturing of primary somatic embryos.Mature somatic embryos with green cotyledons could germinate and develop plantlets.The germination frequencies of somatic embryos from NZ188 and SC8 could reach more than 80%.Using the pieces of somatic cotyledons as explants,secondary SE and SO could be induced after culturing on SE and SO induction media,respectively.The SE frequencies from somatic cotyledons of all tested cultivars achieved more than 77%.On SO medium,shoot meristems and adventitious shoots were developed after 2 weeks.The induction frequencies were between 34.4% and 72.2% among tested cultivars,and the performance of NZ188,NZ199 and SC124 were better than that of other cultivars.Adding ethylene inhibitor AgNO3 to the regeneration medium significantly reduced callus formation and improved the development of shoots,including regeneration rate and average shoot number per explant.AgNO3 at 4 mg/L has been considered the most suitable concentration to be used in this study.The shoots regenerated from AgNO3-containing medium grew much better and were easy for planting into soil.

关 键 词：木薯 体细胞胚胎发生 芽器官发生 硝酸银 植株再生 

分 类 号：S682.33[农业科学—观赏园艺]