草地早熟禾新格莱德胚性愈伤组织原生质体培养及植株再生的研究

草业学报 ›› 2010, Vol. 19 ›› Issue (2): 55-60.

草地早熟禾新格莱德胚性愈伤组织原生质体培养及植株再生的研究

赵小强^1,2,3,马晖玲^1,2,3*,林栋^1,2,3,周万海^1,2,3,吴翔^1,2,3

1.甘肃农业大学草业学院,甘肃兰州 730070;
2.草业生态系统教育部重点实验室, 甘肃兰州 730070;

3.中－美草地畜牧业可持续研究中心,甘肃兰州 730070

收稿日期:2009-02-23 出版日期:2010-02-25 发布日期:2010-04-20
作者简介:赵小强(1985-),男,甘肃武山人,在读硕士。E-mailwushan2002zxq@163.com
基金资助:
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)和甘肃省教育厅项目(0702-03)资助。

Culture and plant regeneration of protoplasts from embryogenic calli of Nuglade (Poa pratensis)

ZHAO Xiao-qiang^1,2,3, MA Hui-ling^1,2,3, LIN Dong^1,2,3, ZHOU Wan-hai^1,2,3, WU Xiang^1,2,3

1.College of Pratacultural Science, Gansu Agricultural University, Lanzhou 730070, China;
2.Key
Laboratory of Grassland Ecosystem, Ministry of Education, Lanzhou 730070, China;
3.Sino-U.S.
Centers for Grazingland Ecosystem Sustainability, Lanzhou 730070, China

Received:2009-02-23 Online:2010-02-25 Published:2010-04-20

摘要/Abstract

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3.0mg/L2,4-二氯苯氧乙酸(2,4-D)、0.5mg/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3.0mg/L2,4-D、0.5mg/L6-BA、100mg/L水解酪蛋白、100mg/L水解乳蛋白、1%蔗糖、0.4mol/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3d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3～5mm时,转入固体培养基MS+3.0mg/L2,4-D+0.5mg/L6-BA和MS+0.5mg/L萘乙酸(NAA)+5.0mg/L6-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。

Abstract: 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.

中图分类号:

赵小强,马晖玲,林栋,周万海,吴翔. 草地早熟禾新格莱德胚性愈伤组织原生质体培养及植株再生的研究[J]. 草业学报, 2010, 19(2): 55-60.

ZHAO Xiao-qiang, MA Hui-ling, LIN Dong, ZHOU Wan-hai, WU Xiang. Culture and plant regeneration of protoplasts from embryogenic calli of Nuglade (Poa pratensis)[J]. Acta Prataculturae Sinica, 2010, 19(2): 55-60.

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