BAO Han,ZHANG Rui,ZHANG Meiling,et al.Establishment of a Regeneration System From Leaf Explant of Stevia rebaudiana[J].Northern Horticulture,2018,42(06):16-22.[doi:10.11937/bfyy.20173097]
甜叶菊叶片外植体再生体系的建立
- Title:
- Establishment of a Regeneration System From Leaf Explant of Stevia rebaudiana
- Keywords:
- Stevia rebaudiana; tissue culture; plant growth regulator; callus; adventitious bud; cluster bud
- 文献标志码:
- A
- 摘要:
- 以甜叶菊品种“中山二号”为试材，首先选用叶片、茎段和茎节3种外植体，在添加3种植物激素萘乙酸(1-Naphthaleneacetic acid,NAA)、苄氨基嘌呤(6-Benzylaminopurine,BAP)和激动素(kinetin,KT)不同浓度组合的MS(murashige and skoog)培养基上培养，明确其对愈伤组织诱导和不定芽分化的影响。以叶片作为外植体，使用激素噻苯隆(N-phenyl-N′-1,2,3-thidiazol-5-yl-urea,TDZ)替换NAA(BAP和KT浓度范围不变)，在不同浓度组合的MS培养基上进行愈伤组织、不定芽和丛生芽诱导，以期建立一套快速、有效的甜叶菊再生体系。结果表明：在培养基中TDZ浓度达到3.0 mg?L-1、BAP 1.0 mg?L-1和KT 0.5 mg?L-1时，诱导的愈伤组织分化的不定芽最多，且随后形成了大量的丛生芽。丛生芽继代到无添加激素的1/2MS培养基上，能够继续生长，分化成茎叶和根，形成完整植株。该再生体系生产周期短、繁殖系数高，可用于生产上大规模快速繁殖和基因转化研究。
- Abstract:
- In order to develop a rapid and efficient method for plant regeneration of Stevia rebaudiana,the segments of leaf,stem and nodalcut from the cultivar ‘Zhongshan No.2’,were used as explants,and the effects of the MS(murashige and skoog) basal medium supplied with three kinds of plant growth regulators,NAA(1-Naphthaleneacetic acid),BAP(6-Benzylaminopurine) and KT(kinetin) in different concentrations on the stages of callus and adventitious bud inducement were determined.Then the leaf was selected as optimal explant,and was cultured on the media that substituted TDZ(N-phenyl-N′-1,2,3-thidiazol-5-yl-urea) for NAA(BAP and KT remained unchanged).The results showed that the most adventitious buds were differentiated from the induced calli on the MS basal medium supplied with 3.0 mg?L-1 TDZ combined with 1.0 mg?L-1 BAP and 0.5 mg?L-1 KT,following a large number of cluster bud sappeared.The cluster buds elongated and differentiated leaves,stem and roots into a complete plantlet.With short production cycle,high propagation coefficient,the regeneration method could be used for rapid mass propagation and gene transformation.
参考文献/References:
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备注/Memo
第一作者简介：包晗（1986-），女，宁夏银川人，硕士，实验师，现主要从事植物组织培养等研究工作。E-mail:18795209024@163com.责任作者：陈任（1963-），男，福建龙海人，博士，教授，现主要从事植物分子生物学与基因工程等研究工作。E-mail:chenren511@163.com.基金项目：宁夏大学引进人才科研启动经费资助项目(BQC2012001)。收稿日期：2017-12-16