小菊悬浮细胞培养与植株再生研究

园艺学报 ›› 2006, Vol. 33 ›› Issue (5): 1021-1026.

小菊悬浮细胞培养与植株再生研究

陈发棣;蒋甲福;郭维明;房伟民;赵宏波

(南京农业大学园艺学院, 江苏南京210095)

收稿日期:2006-01-25 修回日期:2006-03-26 出版日期:2006-10-25 发布日期:2006-10-25

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

Chen Fadi; Jiang Jiafu;GuoWeiming;Fang Weimin;Zhao Hongbo

(College of Horticulture, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China)

Received:2006-01-25 Revised:2006-03-26 Online:2006-10-25 Published:2006-10-25

摘要/Abstract

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+ 1.0 mg·L ^-1 2, 4-D + 0.2 mg·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 + 0.5 mg·L ^{- 1} 2, 4-D + 0.2 mg·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(0～2 d) 、对数生长期(2～10 d) 和停滞期(10 d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 + 0.2 mg·L ^{- 1} KT + 0.2 mg·L ^{- 1} 2, 4-D上植板率最高; 4℃低温2 h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0.2 cm的愈伤组织, 将其转到MS + 2 mg·L ^-1 6-BA + 0.1 mg·L ^{- 1} NAA培养基后, 继代4次, 分化后出芽; 分化芽在培养基MS + 0.5 mg·L ^-1 NAA上诱导生根, 形成小植株。

关键词: 小菊, 悬浮细胞培养, 植株再生

Abstract: 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^-16-benzylamino purine ( 6-BA) could efficiently induce rapid-growing and loose calli. Obtained calli were subsequently cultured 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 cultured 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 ofmedium decreased rapidly during the culture 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 p lant efficiency decreased with increasing of subculture times. Calli with a diameter of 0.2 cm were obtained in one month. When theywere transferred to MS medium containing 2.0 mg·L ^-1 6-BA and 0.1 mg·L ^-1 naphthaleneacetic (NAA) and subcultured for 4 times, shoots differentiated from the calli. Rooting plantlets were induced on MS with 0.5 mg·L ^-1 NAA.

Key words: Dendranthema ×grandiflorum, Cell suspension culture, Plant regeneration

陈发棣;蒋甲福;郭维明;房伟民;赵宏波. 小菊悬浮细胞培养与植株再生研究[J]. 园艺学报, 2006, 33(5): 1021-1026.

Chen Fadi;Jiang Jiafu;GuoWeiming;Fang Weimin;Zhao Hongbo. Studies on Cell Suspension Culture and Plant Regeneration of Dendranthema×grandiflorum with Small Inflorescences[J]. ACTA HORTICULTURAE SINICA, 2006, 33(5): 1021-1026.

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