植物学报 ›› 2015, Vol. 50 ›› Issue (5): 623-627.doi: 10.11983/CBB14174

• 技术方法 • 上一篇    下一篇

以黄花乌头发根为外植体的再生培养体系建立

刘莉莉, 卢淑波, 徐佳萍, 张庆田, 李昌禹*()   

  1. 中国农业科学院特产研究所, 长春 130112
  • 收稿日期:2014-09-22 接受日期:2015-03-30 出版日期:2015-09-01 发布日期:2015-10-09
  • 通讯作者: 李昌禹 E-mail:lcy_lcy2002@163.com
  • 作者简介:

    ? 共同第一作者

  • 基金资助:
    吉林省科学技术厅自然科学基金(No.201115131)

Establishment of Plant Regeneration System from Hairy Roots of Aconitum coreanum

Lili Liu, Shubo Lu, Jiaping Xu, Qingtian Zhang, Changyu Li*   

  1. Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China
  • Received:2014-09-22 Accepted:2015-03-30 Online:2015-09-01 Published:2015-10-09
  • Contact: Li Changyu E-mail:lcy_lcy2002@163.com
  • About author:

    ? These authors contributed equally to this paper

摘要:

为获得黄花乌头(Aconitum coreanum)发根再生植株, 将其发根在添加0.5-5.0 mg·L-1 6-BA及0-0.5 mg·L-1 NAA的1/2MS固体培养基上进行光诱导培养, 发根经脱分化形成愈伤组织, 然后形成芽及芽丛, 最终获得发根再生植株。结果表明, 愈伤组织及不定芽分化的最适培养基为1/2MS+2.0 mg·L-1 6-BA+0.2 mg·L-1 NAA+3%蔗糖; 不定芽生根最适培养基为1/2MS+0.5 mg·L-1 6-BA+0.1 mg·L-1 NAA+5 mg·L-1 GA+3%蔗糖。

Abstract:

The hairy roots of Aconitum coreanum were used as explants to induce callus via differentiation in 1/2MS medium with 0.5-5 mg·L-1 6-BA and 0-0.5 mg·L-1 NAA, then regeneration plantlets were induced from the callus. A whole transgenetic plantlet was obtained. The optimal medium for inducing callus and adventitious buds was 1/2MS+2.0 mg·L-1 6-BA+0.2 mg·L-1 NAA+3% sugar. The optimal medium for inducing roots was 1/2MS+0.5 mg·L-1 6-BA+0.1 mg·L-1 NAA+5 mg·L-1 GA+3% sugar.

图1

黄花乌头发根诱导植株再生 (A) 发根; (B) 基部膨大的发根; (C) 愈伤组织; (D) 芽丛; (E) 完整植株; (F) 白化苗; (G) 再生根。Bar=5 mm"

表1

不同激素配比对诱导黄花乌头产生愈伤组织及不定芽的影响"

No. of
medi- um
Phytohormone
composition (mg·L-1)
Color change of hairy roots Callus
induced
Adventitious buds induced
NAA 6-BA
M1 0 0.5 10% turned green - -
M2 0 1.0 30% turned green - -
M3 0 2.0 10% could see green spots - -
M4 0 4.0 5% could see green spots - -
M5 0 5.0 5% could see green spots - -
M6 0.1 0.5 50% turned green - 3%
M7 0.2 0.5 60% turned yellow-green 20% Few adventitious buds were induced
M8 0.5 0.5 40% turned yellow-green 20% -
M9 0.1 1.0 90% turned yellow-green - 10%
M10 0.2 1.0 40% turned yellow-green, 20% turned green 25% Few adventitious buds were induced
M11 0.5 1.0 50% turned yellow-green 30% Few adventitious buds were induced
M12 0.1 2.0 30% turned yellow-grown 30% 30%
M13 0.2 2.0 40% turned yellow-grown, 20% turned green - -
M14 0.5 2.0 40% turned yellow-green Few callus
were obtained
-
M15 0.1 4.0 30% turned green - -
M16 0.2 4.0 30% turned yellow-green 20% 40%
M17 0.5 4.0 40% turned yellow-green 20% -
M18 0.1 5.0 20% turned green - -
M19 0.2 5.0 50% turned yellow-green, 50% turned green 50% Few adventitious buds were induced
M20 0.5 5.0 50% turned yellow-green 20% 20%

表2

植物材料在原培养基上继代培养1次后的生长状态"

No. of medium Phytohormone composition (mg·L-1) Description of hairy root growing and differentiation status
NAA 6-BA
M6 0.1 0.5 The material turned albino and died
M7 0.2 0.5 30% of the material turned brown, 10% turned green, 30% turned yellow slim hairy roots, 10% produced cluster buds
M8 0.5 0.5 80% turned yellow slim hairy roots, the base section of hairy roots became enlargement and 10% base section turned green
M9 0.1 1.0 The material turned albino and died
M10 0.2 1.0 The material turned albino and died
M11 0.5 1.0 30% base section of hairy roots turned green, 20% new produced hairy roots grew long and thin, others were short and thick
M12 0.1 2.0 30% of the material turned green, 30% kept yellow-green and enlargement
M13 0.2 2.0 The material turned albino and died
M14 0.5 2.0 50% of the material turned green and was going to be buds, hairy roots grew thick, 40% hairy roots were becoming brown
M15 0.1 4.0 40% of the material turned green and some were going to be buds, hairy roots were enlargement and strong, 50% were becoming brown
M16 0.2 4.0 The material turned albino and died
M17 0.5 4.0 The material turned albino and died
M18 0.1 5.0 The material turned albino and died
M19 0.2 5.0 40% of the material turned green, 20% kept yellow-green and grew strongly, 20% turned brown
M20 0.5 5.0 50% of the material turned green and grew strongly, 10% became cluster plantlets, 20% turned brown

表3

黄花乌头再生苗诱导生根适宜培养基"

Phytohormone composition
(mg·L-1)
No. of root
0.1 0.5 1.0 1.5
IBA 1 2 6 6
NAA 5 3 1 0
IAA 0 1 3 4
0.5 mg·L-1 6-BA+NAA 4 3 2 1
0.5 mg·L-1 6-BA +0.5 mg·L-1 GA+ NAA 8 3 1 0
1 冯欢, 易姝利, 谢佳恒, 雷梦琦, 黄萱 (2014). 微型月季愈伤组织诱导及植株再生. 植物学报 49, 595-602.
2 侯丽丽, 施和平, 余武, 曾宝强, 周卓辉 (2014). 烟草毛状根多倍体诱导及其植株再生. 生物工程学报 30, 581-594.
3 李昌禹, 王英平, 卢淑波, 张庆田, 孔祥义, 徐佳萍, 艾军 (2012). 黄花乌头转基因苗及其获得方法. 中国专利. 2011- 10317204.4. 2012-06-20.
4 李昌禹, 王英平, 杨福合, 卢淑波, 臧埔, 孔长伟, 孔祥义 (2006). 黄花乌头发根及其获得方法. 中国专利. 2006100- 89523.3. 2006-12-27.
5 李昌禹, 王英平, 臧埔, 卢淑波, 宋婷婷 (2005). 黄花乌头叶片诱导植株再生研究. 特产研究 27(2), 34-36.
6 施和平, 朱远锋, 王蓓, 孙蒋兵, 黄胜琴 (2014). 香石竹毛状根诱导、离体培养及其植株再生. 生物工程学报 30, 1742-1750.
7 中国科学院中国植物志编辑委员会 (1979). 中国植物志(第27卷). 北京: 科学出版社. pp. 323.
8 Bai YZ, Li CY, Zhao JH, Zheng PH, Li YL, Pan YQ, Wang YP (2013). A high yield method of extracting alkaloid from Aconitum coreanum by pulsed electric field.Chromatographia 76, 635-642.
9 ?
10 ? … show all 7 hide
11 Li CY, Lu SB, Kong XY, Liu LL, Wang YP (2013). Optimization of suspension culture conditions for hairy roots of Aconitum coreanum (Levl.) Raipaics.Agric Biotechnol 2(4), 5-7.
No related articles found!
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] 戴锡玲;曹建国;王全喜*. 水蕨孢子壁的形成和发育[J]. 植物学报, 2008, 25(01): 72 -79 .
[2] 刘德立. 植物热激蛋白及其功能[J]. 植物学报, 1996, 13(01): 14 -19 .
[3] 丁承强 马丹 王绍华 丁艳锋. 水稻蛋白质组双向电泳优化流程及方法[J]. 植物学报, 2011, 46(1): 67 -73 .
[4] 宋克敏. 植物的磷营养:磷酸盐运转系统及其调节[J]. 植物学报, 1999, 16(03): 251 -256 .
[5] 杨洪强 接玉玲 李军. 植物根源逆境信使及其产生和传输[J]. 植物学报, 2002, 19(01): 56 -62 .
[6] 李辉, 张光灿, 谢会成, 许景伟, 李传荣, 孙居文. 苯酚废水对垂柳叶片光合生理参数的影响[J]. 植物学报, 2016, 51(1): 31 -39 .
[7] 刘林德 王仲礼 田国伟 申家恒. 刺五加胚与胚乳发育的研究[J]. 植物学报, 1996, 13(专辑): 97 -98 .
[8] 刘晓梅, 方建, 张婧, 林吴颖, 樊廷录, 冯虎元. 长期施肥对麦田土壤微生物垂直分布的影响[J]. 植物生态学报, 2009, 33(2): 397 -404 .
[9] 王仁卿. 山东半岛和辽东半岛植被的比较研究[J]. 植物生态学报, 1984, 8(1): 41 -51 .
[10] 宋永昌. 第二届欧洲生态学学术讨论会和第十届德国生态学年会[J]. 植物生态学报, 1981, 5(2): 151 -166 .