草地早熟禾午夜Ⅱ号原生质体培养及植株再生

doi:10.11733/j.issn.1007-0435.2010.01.020

›› 2010, Vol. 18 ›› Issue (1): 103-107.DOI: 10.11733/j.issn.1007-0435.2010.01.020

草地早熟禾午夜Ⅱ号原生质体培养及植株再生

马晖玲, 赵小强, 白小明

甘肃农业大学草业学院, 草业生态系统教育部重点实验室, 中-美草地畜牧业可持续研究中心, 甘肃, 兰州, 730070

收稿日期:2009-04-30 修回日期:2009-12-14 出版日期:2010-02-15 发布日期:2010-02-15
作者简介:马晖玲(1966- ),女,回族,甘肃兰州人,博士,教授,主要从事以细胞工程和基因工程手段改良草地植物品种特性的研究,E-mail:mahl@gsau.edu.cn
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);甘肃省教育厅项目(0702-03)

Protoplast Culture and Plant Regeneration of Midnight Ⅱ(Poa Pratensis L.)

MA Hui-ling, ZHAO Xiao-qiang, BAI Xiao-ming

Key laboratory of Grassland Ecosystem of Ministry of Education & Sino-US Centers for Grazingland Ecosystem Sustainability, Pratacultural College, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China

Received:2009-04-30 Revised:2009-12-14 Online:2010-02-15 Published:2010-02-15

摘要/Abstract

摘要： 对草地早熟禾(Poa Pratensis L.)品种-午夜Ⅱ号进行了原生质体培养及其植株再生的研究。以午夜Ⅱ号成熟种子诱导的松软胚性愈伤组织为材料,利用酶解法分离出大量有活力的原生质体。原生质体经培养,持续分裂形成愈伤组织,并分化出再生绿苗和白化苗。研究了不同酶液组成和酶解时间对原生质体游离的影响。结果表明:采用继代培养8-10 d的胚性愈伤组织在1.0%纤维素酶+1.0%离析酶+0.5%果胶酶+0.3%崩溃酶条件下,酶解14-16 h可得到产量和活力较高的原生质体。原生质体以2.0×10⁵-5.0×10⁵个/mL的植板密度,采用液体浅层法在附加1.0 mg/L 2,4-D、0.3 mg/L 6-BA、100 mg/L水解酪蛋白、100 mg/L水解乳蛋白、1.0%蔗糖、0.4 mol/L甘露醇的KM₈P培养基中培养,可形成较小的愈伤组织。愈伤组织经过增殖在分化培养基上(MS+0.5mg/LNAA+2.0 mg/L 6-BA)可诱导芽、根的形成,再生出完整植株。

关键词: 草地早熟禾, 午夜Ⅱ号, 原生质体培养, 植株再生

Abstract: The friable calli induced from mature seeds of Poa pratensis L.cv.Midnight II were used for protoplast preparation and the protoplasts were isolated using enzyme digestion.Calli appeared after sustained protoplast divisions in culture medium and the green shoot and albino plantlets were obtained from the protocalli on differentiated medium.The effects of different enzyme composition and enzyme digestion time on protoplast isolation were discussed in this paper.The results show that it was available to harvest high yield and viabilities of protoplasts from embryogenic calli with treatment of digestion in 1.0% Cellulase Onozuka R-10,1.0% Macerozyme R-10,0.3% Driselase,and 0.5% Pectolase Y-23 in 8 to 10 d.It was better to digest for 14-16 h in enzyme digestion solution mentioned above.Calli formed on KM₈P medium supplemented with 1.0 mg/L 2,4-D,0.3 mg/L 6-BA,100 mg/L casein hydrolysate,100 mg/L lactablumin hydrolysate,1.0%(W/V) sucrose,and 0.4 mol/L mannitol at plating density of 2.0×10⁵-5.0×10⁵ mL^-1.The protoplast-derived calli developed shoots and roots on differentiation media(MS+0.5 mg/L NAA+2.0 mg/L 6-BA) after further subculture,and then complete plants were obtained.

Key words: Poa pratensis, Midnight II, Protoplast culture, Plant regeneration

中图分类号:

Q813.11

马晖玲, 赵小强, 白小明. 草地早熟禾午夜Ⅱ号原生质体培养及植株再生[J]. , 2010, 18(1): 103-107.

MA Hui-ling, ZHAO Xiao-qiang, BAI Xiao-ming. Protoplast Culture and Plant Regeneration of Midnight Ⅱ(Poa Pratensis L.)[J]. , 2010, 18(1): 103-107.

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草地早熟禾午夜Ⅱ号原生质体培养及植株再生

Protoplast Culture and Plant Regeneration of Midnight Ⅱ(Poa Pratensis L.)

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