[1]杨利艳,冉慧,乔白露,等.基于绿色荧光蛋白观察的花粉介导转化泡桐初探[J].北方园艺,2014,38(09):117-120.
　YANG Li-yan,RAN Hui,QIAO Bai-lu,et al.Preliminary Study on Paulownia Transformation via Pollen-mediated Transformation Approach Based on GFP Observation[J].Northern Horticulture,2014,38(09):117-120.

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基于绿色荧光蛋白观察的花粉介导转化泡桐初探

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第38卷期数: 2014年09期页码: 117-120 栏目: 出版日期: 2014-05-15

Title:: Preliminary Study on Paulownia Transformation via Pollen-mediated Transformation Approach Based on GFP Observation

文章编号:: 1001-0009（2014）09-0117-04

作者:: 杨利艳¹; 冉慧¹; 乔白露¹; 孙毅²; 3; (1.山西师范大学生命科学学院，山西临汾 041004;
2.山西省农业科学院生物技术研究中心,山西太原 030031;
3.农业部黄土高原作物基因资源与种质创制重点实验室,山西太原 030031

Author(s):: YANG Li-yan¹; RAN Hui¹; QIAO Bai-lu1; SUN Yi²; 3; 1.College of Life Science,Shanxi Normal University,Linfen,Shanxi 041004;
2.Biotechnology Research Center,Shanxi Academy of Agricultural Sciences,Taiyuan,Shanxi 030031;
3.Key Laboratory of Crop Gene Resources and Germplasm Enhancementon Loess Plateau,Ministry of Agriculture,Taiyuan,Shanxi 030031

关键词:: 泡桐; 超声波; 花粉介导转化; 绿色荧光蛋白（GFP）

Keywords:: Paulownia; ultrasonication; pollen-mediated transformation; green fluorescence protein (GFP)

分类号:: S 792.43

文献标志码:: A

摘要:: 以泡桐花粉为受体，在超声波的作用下将含有绿色荧光蛋白（GFP）基因的质粒与花粉共处理，利用荧光显微镜对GFP基因在花粉粒及花粉管中的表达进行了示踪观察。结果表明：100 g/L 的蔗糖溶液是泡桐花粉较适宜的体外培养液；花粉粒有较强烈的自发荧光，因此不能根据花粉粒荧光来确定GFP基因是否表达；处理组花粉管较对照呈现强烈的绿色荧光，表明GFP基因在花粉管中表达。该试验首次利用泡桐花粉对GFP基因的表达观察，初步证明了这一转基因方法对泡桐花粉转化的可行性。

Abstract:: Taking Paulownia pollen as receptor,the fresh Paulownia pollen was treated under ultrasonication mixed with plasmid DNA harboring green fluorescence protein (GFP) gene,the expressions of GFP gene in pollen grains,pollen tubes were observed under a fluorescence microscope.The results showed that 100 g/L sucrose was the suitable concentration for in vitro pollen germination.Pollen grains had strong green autoflorescent background,so GFP gene was not a reliable reporter for tracing the transformation of exotic genes in pollen.Pollen tubes in the treatment group produced strong green florescence compared to the control which meant the GFPgene expression in pollen tubes.The present study preliminarily provided evidence for Paulownia genetic transformation via pollen-mediated transformation approach.

参考文献/References:

[1]钟启宏,孔繁瑞,孙威,等.通过花粉管途径建立泡桐转化系统的初探[J].遗传,1994,16(6):16-19.

[2]施士争,倪善庆.泡桐组织培养系统性研究初探[J].江苏林业科技,1995,22(3):20-22.

[3]FanG Q,Zhai X Q,Zhai C J,et al.Callus induction from leaves of different?Paulownia?species and its plantlet regeneration[J].Journal of ForestryResearch,2001,12(4):209-214.

[4]范国强,彭海风,翟晓巧,等.泡桐叶片蛋白质多态性及其聚类分析[J].植物学通报,2001,18(6):739-774.

[5]范国强,翟晓巧,蒋建平,等.不同种泡桐叶片愈伤组织诱导及其植株再生[J].林业科学,2002,38(1):29-35.

[6]范国强,翟晓巧,马新业.两种泡桐叶片体细胞胚胎发生及植株再生[J].核农学报,2005,19(4):274-278.

[7]范国强,董占强,李峰稳,等.光周期对泡桐叶片离体植株再生影响研究[J].西北植物学报,2007,27(1):104-109.

[8]Kalaycioglu H,Deniz I,Hiziroglu S.Some of the properties of particleboard made from Panlownia[J].Journal of Wood Science,2005,51:410-414.

[9]刘飞,范国强,董占强.泡桐离体开花培养系统的建立[J].林业科学,2007,43(12):56-63.

[10]孙毅,王景雪,崔贵梅.超声波处理花粉介导植物基因转化方法[P].中国：ZL991211529，20060329.

[11]Wang J X,Sun Y,Cui G M,et al.Transgenic maize plants obtained by pollen-mediated transformation [J].Acta Botanica Sinica,2001,43：275-279.

[12]Wang W Q,Wang J X,Yang C Q,et al.Pollen-mediated transformation of Sorghum bicolor?plants [J].Biotechnology Applied Biochemistry,2007,48:79-83.

[13]Wang J X,Li Y H,Liang C.Recovery of transgenic plants by pollen mediated transformation in?Brassica juncea[J].Transgenic Research,2008,17:417-424.

[14]Eapen S.Pollen grains as a target for introduction of foreign genes into plants:an assessment [J].Physiology and Molecular Biology of Plants,2011,17:1-8.

[15]Sambrook J,Maniatis T,Fritsch E F.Molecular Cloning:A Laboratory Manual (2ndedition)[M].New York:Cold Spring Harbor Labrotory Press，1989.

[16]陈志远.泡桐属（Paulownia）花粉形态学的初步研究[J].武汉植物学研究,1983,1(2):143-147.

[17]ChenX,Equi R,Baxter H,et al.?A high-throughput transient gene expressionsystem for switchgrass (Panicum virgatumL.) seedlings [J].Biotechnologyfor Biofuels,2010,3:1-10.

[18]Ottenschlager I,Barinova I,Voronon V,et al.Green fluorescent protein (GFP) as a marker during pollen development [J].Transgenic Research,1999,4:279-294.

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备注/Memo

第一作者简介：杨利艳（1972-），女，博士，副教授，现主要从事植物基因工程等研究工作。E-mail：yangswallow163@163.com.

责任作者：孙毅（1953-），男，博士，研究员，现主要从事植物育种及基因工程等研究工作。E-mail:sunyi692003@163.com.

基金项目：山西省回国留学人员科研资助项目(2011-061)；国家农业部转基因生物新品种培育重大专项资助项目(2013ZX08003-001)；国家自然科学基金资助项目（31240081）；山西省科技攻关资助项目(20110311009)。

更新日期/Last Update: 2014-08-13