小菊悬浮细胞培养与植株再生研究  被引量:11

Studies on Cell Suspension Culture and Plant Regeneration of Dendranthema×grandiflorum with Small Inflorescences

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作  者:陈发棣[1] 蒋甲福[1] 郭维明[1] 房伟民[1] 赵宏波[1] 

机构地区:[1]南京农业大学园艺学院,江苏南京210095

出  处:《园艺学报》2006年第5期1021-1026,共6页Acta Horticulturae Sinica

基  金:江苏省科技厅高技术研究项目(BG2003305;BG2004310);上海市农委重点攻关项目〔农科攻字(2004)第3-1〕;江苏省高技术产业化项目(JH02-086);江苏省农业三项工程项目〔SX(2003)065〕

摘  要:以小菊‘七月红’品种为试材,进行了细胞悬浮培养及植株再生诱导的探讨。结果表明,MS+1.0mg·L^-1 2,4-D+0.2mg·L^-1 6-BA培养基适合从试管苗茎段有效诱导生长迅速、质地疏松的愈伤组织。获得的愈伤组织在MS+0.5mg·L^-1 2,4-D+0.2mg·L^-1 6-BA液体培养基中振荡培养,建立细胞悬浮培养体系。悬浮细胞的生长曲线呈上升的半抛物线型,细胞生长大致分缓慢生长期(0~2d)、对数生长期(2~10d)和停滞期(10d以后);随着悬浮细胞生长,培养液pH值迅速下降。悬浮细胞固液双层培养表明,在培养基MS+0.2mg·L^-1 KT+0.2mg·L^-1 2,4-D上植板率最高;4℃低温2h处理能促进悬浮细胞生长,提高植板率;随着继代次数增加植板率呈下降趋势。培养1个月后,形成了直径约0.2cm的愈伤组织,将其转到MS+2mg·L^-1 6-BA+0.1mg·L^-1 NAA培养基后,继代4次,分化后出芽;分化芽在培养基MS+0.5mg·L^-1 NAA上诱导生根,形成小植株。Chrysanthemum ‘ Qiyuehong' with small inflorescence was employed for study of cell suspension and plant regeneration. The results showed that MS (Murashige and Skoog) medium with supplement of 1.0 mg · L^-1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0. 2 mg · L^-1 6-benzylamino purine (6-BA) could efficiently induce rapid-growing and loose calli. Obtained calli were subsequently cuhured in liquid MS medium with 0. 5 mg ·L^-1 2,4-D and 0. 2 mg · L^-1 6-BA for establishing cell suspension system. The growth of suspension cuhured cells showed a parabola curve, mainly characterized by lagged growth phase (0 -2 d), logarithmic growth phase (2 -10 d) and stationary growth phase (10 d later), respectively. In addition, pH value of medium decreased rapidly during the cuhure process. Highest plant efficiency was obtained on MS medium with 0. 2 mg · L^-1 kinetin (KT) and 0. 2 mg· L^-1 2,4-D. Low temperature treatment at 4% for 2 hours could enhance the growth of cell, and increase the planting efficiency as well. However, the plant efficiency decreased with increasing of subcuhure times. Calli with a diameter of 0. 2 cm were obtained in one month. When they were transferred to MS medium containing 2. 0 mg ·L^-1 6-BA and 0. 1 mg · L^-1 naphthaleneacetic (NAA) and subcuhured for 4 times, shoots differentiated from the calli. Rooting plantlets were induced on MS with 0. 5 mg· L^-1 NAA.

关 键 词:小菊 悬浮细胞培养 植株再生 

分 类 号:S682.11[农业科学—观赏园艺]

 

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