草地早熟禾农杆菌介导法基因转化条件

doi:10.11733/j.issn.1007-0435.2005.01.010

›› 2005, Vol. 13 ›› Issue (1): 39-42,70.DOI: 10.11733/j.issn.1007-0435.2005.01.010

草地早熟禾农杆菌介导法基因转化条件

佘建明¹, 张保龙¹, 何晓兰¹, 陈志一², 倪万潮¹

1. 江苏省农业科学院农业生物遗传生理研究所, 南京, 210014;
2. 浙江绍兴白云建设有限公司草业研究所, 新昌, 312500

收稿日期:2003-12-11 修回日期:2004-02-23 出版日期:2005-02-15 发布日期:2005-02-15
作者简介:余建明(1945- ),男,江苏苏州人,研究员,主要从事作物农业生物技术研究,已发表论文40余篇,E-mail:SheJM101@hotmail.com.
基金资助:
浙江省科技厅科研项目(2003C32067);江苏省农业科学院重点领域研究项目(6320301)

A Study on the Conditions of Genetic Transformation into Poa pratensis L. by Agrobacterium Tumefaciens

SHE Jian-ming¹, ZHANG Bao-long¹, HE Xiao-lan¹, CHEN Zhi-yi², NI Wan-chao¹

1. Institute of Agrobiological Genetics and Physiology, Jiangsu Academy of Agricultural Sciences, Nanjing, Jiangsu Province 210014, China;
2. Institute of Forage Industry, Zhejiang Shaoxing Baiyun Construction Co. Ltd, Xinchang, Zhejiang Province 312500, China

Received:2003-12-11 Revised:2004-02-23 Online:2005-02-15 Published:2005-02-15

摘要/Abstract

摘要： 以大青山草地早熟禾(Poapratensis L.cv.Daqingshan)和超级伊克利草地早熟禾(PoapratensisL.cv.Totaleclipse)成熟种子诱导产生的胚性愈伤组织为材料,比较抗生素及筛选剂对愈伤组织生长和绿苗分化的影响.结果表明:500mg/L的羧苄青霉素或头孢霉素对愈伤组织的生长与对照间差异不显著;羧苄青霉素可提高愈伤组织绿苗分化率,而头孢霉素则有抑制作用;2个供试品种G-418的筛选浓度分别为50和70mg/L时,愈伤组织的褐化率达100%;在幼苗分化期,G-418浓度为20mg/L时,2个供试品种的试管苗均为白化苗;在观察到愈伤组织周围的培养基上显现菌落时,立即转移到筛选培养基上为宜.用农杆菌介导法获得了转苏云金芽孢杆菌杀虫晶体蛋白(B.t)基因植株.

关键词: 草地早熟禾, 胚性愈伤组织, 根癌农杆菌, B.t基因, 转基因植株

Abstract: Using the embryogenic calli induced from the mature seeds of Poa pratensis L. cv. Daqingshan and Total eclipse as recipient materials, authors of the paper studied the effect of carbenicillin, cefotaxime, and geneticin (G-418) on the callus growth and shoot regeneration. The result shows that the effect of 500 mg/L carbenicillin or cefotaxime to the growth of calli matches that of the control. Carbenicillin could increase the rate of shoot regeneration of calli, while cefotaxime checks it. The calli of the two species, “Daqingshan” and “Total eclipse”, screened separately with 50 mg/L and 70 mg/L of G-418 both turn brown. During the shoot regeneration period, 20 mg/L of G-418 turns all the shoots white. The time needed for culturing the embryogenic callus and agrobacterium tumefaciens together is determined by the appearance of colonies of the latter that could be detected with naked eyes. The callus should be transferred immediately to the screening medium. The recombinant plasmid carrying B.t gene and NPT Ⅱ gene are introduced into callus of “Total eclipse” by means of agrobacterium-mediated transformation. Transgenic plants confirmed by PCR analysis are thus obtained.

Key words: Poa pratensis, Embryogenic callus, Agrobacterium tumefaciens, B.t. gene, Transgenic plant

中图分类号:

Q813.1
S812

佘建明, 张保龙, 何晓兰, 陈志一, 倪万潮. 草地早熟禾农杆菌介导法基因转化条件[J]. , 2005, 13(1): 39-42,70.

SHE Jian-ming, ZHANG Bao-long, HE Xiao-lan, CHEN Zhi-yi, NI Wan-chao. A Study on the Conditions of Genetic Transformation into Poa pratensis L. by Agrobacterium Tumefaciens[J]. , 2005, 13(1): 39-42,70.

参考文献

[1] Wang Z Y, Takamizo T, Iglesias V A, et al. Transgenic plants of tall fescue (Festuca arundinacea Schreb. ) obtained by direct gene transfer to protoplasts [J]. Bio/Technology, 1992, 10:691-696
[2] Dalton S J, Bettany A J E, Timms E, et al. The effect of selection pressure on transformation frequency and copy number in transgenic plants of tall fescue (Festuca arundinacea Schreb. )[J]. Plant Sci., 1995, 108:63-70
[3] Spangenberg G, Wang Z Y, Nagel J, et al. Protoplast culture and generation of transgenic plants in red fescue (Festuca rubra L. )[J]. Plant Sci. , 1994, 97:83-94
[4] Lee L, Laramore C L, Day P, et al. Transformation and regeneration of creeping bentgrass (Agrostis palustris Huds. )protoplasts[J]. Crop Sci. , 1996, 36:401-406
[5] Wang G R, Binding H, Posselt U K. Fertile transgenic plants from direct gene transfer to protoplasts of Lolium perenne L. and Lolium multiflorum Lam. [J]. J. Plant Physiol, 1997, 151: 83-90
[6] Dalton S J, Bettany A J E, Timms E, et al. Transgeic plants of Lolium multiflorum, Lolium perenne, Festuca arundinacea and Agrostis stolonifera by silicaon carbide fibre-mediated transformation of cell suspension cultures[J]. Plant Sci. , 1998, 132: 31-43
[7] Zhong H, Bolyard M G, Srinivasan C, et al. Transgenic plants of turfgrass (Agrostis palustris Huds) from microprojectile bombardment of embryogenic callus[J]. Plant Cell Rep, 1993, 13:1-6
[8] Spangenberg G, Wang Z Y, Wu X L, et al. Transgenic tall fescue (Festuca arundinacea) and red fescue (F. rubra) plant from microprojectile bombardment of embryogenic suspension cells[J]. J. Plant Pysiol. , 1995, 145:693-700
[9] Spangenberg G, Wang Z Y, Nagel J, et al. Transgenic perennial ryegrass (Lolium perenne) plants from microprojectile bombardment of embryogenic suspension cells[J]. Plant Sci. , 1995, 108:209-217
[10] Ye X, Wang Z Y, Wu X, et al. Transgenic Italian ryegrass(Lolium multiflorum) plants from microprojectile bombardment of embryogenic suspension cells[J]. Plant Cell Rep. , 1997, 16:379-384
[11] Meyer W, Zhang G, Lu S, et al. Transformation of Kentucky bluegrass (Poa pratensis L. ) with betaine aldehyde dehydrogenase gene for salt and drought tolerance[A]. Annual Meetings Abstracts-American Society of Agronomy, Crop Science Society of America, Soil Science ociety of American[C]. Minneapolis Minnesota. November 5-9, 2000.
[12] Liang C M, Hwan K D. Agrobacterium-mediated transformation of Korean lawngrass (Zoysia japonica)[J]. J. Korean Society Horticultural Science, 2000, 41:455-458
[13] Chai B F, Liang A H, Wang W, et al. Agrobacterium-mediated transformation of Kentucky bluegrass[J]. Acta Botanica Sinica,2003, 45(8) :966-973
[14] Murashige T, Skoog F. A revised medium from rapid growth and bioassays with tobacco tissue cultures[J]. Physiol Plant, 1962,15:473-497
[15] Gamborg O L, Miller R A, Ojima K. Nutrient requirements of suspension cultures of soybean root cells [J]. Exp. Cell Res. ,1968, 50:151-158
[16] Pich U, Schubert I. Miniprep method for isolation of DNA from plants with a high content of polyphenolics [J]. Nucleic Acids Research, 1993, 21 (14): 28-33
[17] 佘建明,张保龙,陈志一,等.草地早熟禾成熟胚离体培养植株再生技术的研究[J].草地学报,2003,11(1):58-62
[18] 黄健秋,卫志明,安海龙,等.农杆菌转化获得转B.t.基因水稻及其生物学鉴定[J].植物生理学报,2000,26(6):519-524

草地早熟禾农杆菌介导法基因转化条件

A Study on the Conditions of Genetic Transformation into Poa pratensis L. by Agrobacterium Tumefaciens

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	左郎, 王树森, 马迎梅, 温苏雅勒图, 白刚, 郁鑫明. 火炬树浸提液对两种草坪草种子萌发的影响[J]. 草地学报, 2021, 29(9): 1927-1933.
[2]	金一锋, 陈阳, 齐欣, 高岩松, 金忠民, 赵清峰, 王琦. 草地早熟禾NRT1/PTR FAMILY 8.3基因的克隆及表达分析[J]. 草地学报, 2021, 29(7): 1397-1405.
[3]	朱琨, 郑桂华, 刘丽杰, 周璐, 李珊珊, 范震宇, 钟玥, 尼尼. 草地早熟禾叶片表皮解剖结构与抗白粉病性的研究[J]. 草地学报, 2021, 29(7): 1430-1435.
[4]	张韦钰, 王春勇, 杜红梅. 富氢水对草地早熟禾耐盐性的影响以及与抗氧化酶活性的关系[J]. 草地学报, 2021, 29(7): 1436-1445.
[5]	朱琨, 刘丽杰, 周璐, 李珊珊, 范震宇, 吴纪一, 尼尼. 外源激素对染白粉菌草地早熟禾叶片膜透性及表面结构的影响[J]. 草地学报, 2021, 29(6): 1210-1216.
[6]	朱瑞婷, 牛奎举, 张娣君, 杨阳, 宋云, 马晖玲. 外源2,4-表油菜素内酯对镉胁迫下草地早熟禾叶绿素代谢的影响[J]. 草地学报, 2021, 29(4): 635-643.
[7]	高娅楠, 韩烈保, 许立新. 乙烯利对干旱胁迫下草地早熟禾抗氧化酶基因表达的影响[J]. 草地学报, 2021, 29(10): 2200-2213.
[8]	张然, 马祥, 朱瑞婷, 牛奎举, 赵春旭, 马晖玲. 青海野生草地早熟禾响应干旱胁迫的代谢通路及转录调控分析[J]. 草地学报, 2020, 28(6): 1508-1518.
[9]	赵春旭, 张然, 牛奎举, 朱瑞婷, 王勇, 马祥, 马晖玲. 青海野生草地早熟禾响应低温胁迫的代谢组学研究[J]. 草地学报, 2020, 28(4): 904-914.
[10]	周彤, 师尚礼, 陈建纲, 吴芳, 宫文龙, 来幸樑, 阿芸, 张成君. 紫花苜蓿与草地早熟禾轮作模式对其农艺性状和营养品质的影响[J]. 草地学报, 2020, 28(3): 720-726.
[11]	赵春旭, 马祥, 董文科, 牛奎举, 张然, 马晖玲. 低温胁迫下不同青海野生草地早熟禾的转录组比较分析[J]. 草地学报, 2020, 28(2): 305-318.
[12]	张金青, 陈金龙, 李凡, 赵炳森, 贾新凤, 马晖玲. 草地早熟禾种子萌发和幼苗生长对铅胁迫的适应性[J]. 草地学报, 2020, 28(1): 130-140.
[13]	鲜靖苹, 王勇, 马晖玲. 外源NO缓解草地早熟禾镉胁迫的转录组分析[J]. 草地学报, 2019, 27(6): 1467-1476.
[14]	鲜靖苹, 王勇, 马晖玲. 一氧化氮信号途径参与草地早熟禾耐镉机制的研究[J]. 草地学报, 2019, 27(6): 1577-1586.
[15]	陈润娟, 白小明, 郑文博, 郭宵, 黄琪, 李昌宁, 卢妮. 甘肃地区11个野生草地早熟禾材料根茎扩展性研究[J]. 草地学报, 2019, 27(5): 1250-1258.