摘要
以草地早熟禾品种——新格莱德成熟种子为供试材料,在含3.0 mg/L 2,4-二氯苯氧乙酸（2,4-D）0、.5 mg/L6-苄氨基嘌呤（6-BA）的MB5培养基中进行胚性愈伤组织诱导的培养。并从生长了7-9个月的胚性愈伤组织中分离出原生质体,将该原生质体置于KM8P培养基（含3.0 mg/L 2,4-D、0.5 mg/L 6-BA、100 mg/L水解酪蛋白、100 mg/L水解乳蛋白、1%蔗糖0、.4 mol/L甘露醇）中进行了液体浅层培养。结果表明,新格莱德原生质体在上述KM8P培养基中培养3 d后出现第1次细胞分裂,2-3周后形成小细胞团,此时添加低渗培养液2-3次,小细胞团持续分裂并形成愈伤组织。当愈伤组织块长至3-5 mm时,转入固体培养基MS＋3.0 mg/L 2,4-D＋0.5 mg/L6-BA和MS＋0.5 mg/L萘乙酸（NAA）＋5.0 mg/L 6-BA上进行培养,使其细胞增殖和分化,且逐步形成完整的植株。
Embryogenic calli were induced from mature seeds of Nuglade（Poa Pratensis） on MB5 medium supplemented with 3.0 mg/L 2,4-D,0.5 mg/L 6-BA.Protoplasts were isolated from the embryogeni calli after subculture for 7-9 months.The protoplasts were cultured on KM8P medium supplemented with 3.0 mg/L 2,4-D,0.5 mg/L 6-BA,100 mg/L casein hydrolysate,100 mg/L lactablumin hydrolysate,1%（W/V）sucrose and 0.4 mol/L mannitol.The first divisions happened after 3 days and small cell clusters appeared after 2 to 3 weeks in the culture medium.The protoplast-derived cells could keep dividing sustainably and forming calli by adding fresh protoplast culture liquids once or twice to lower osmotic pressure.The regenerated calli,3 to 5 mm in diameter,were transferred to solid medium（MS medium supplements with 3.0 mg/L 2,4-D,0.5 mg/L 6-BA then MS medium supplements with 0.5 mg/L NAA,5.0 mg/L 6-BA） to carry out subculture for cell propagation,differentiation,then formation of complete plants.
出处
《草业学报》
CSCD
北大核心
2010年第2期55-60,共6页
Acta Prataculturae Sinica
基金
甘肃农业大学草业学院草业科学国家级重点学科学术骨干科研项目暨草业生态系统教育部省部共建重点实验室资助项目(CY-GG-2006-10)
甘肃省教育厅项目(0702-03)资助
