[1]秦正睿,孙磊,李宏宇.利用注射叶片法快速鉴定大豆GmCRY1和GmCRY2基因功能[J].大豆科学,2012,31(01):13-16.[doi:10.3969/j.issn.1000-9841.2012.01.004]
　QIN Zheng-rui,SUN Lei,LI Hong-yu.Rapid Identification of GmCRY1 and GmCRY2 Function by Leaf Injection in Soybean[J].Soybean Science,2012,31(01):13-16.[doi:10.3969/j.issn.1000-9841.2012.01.004]
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利用注射叶片法快速鉴定大豆GmCRY1和GmCRY2基因功能

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第31卷 期数: 2012年01期 页码: 13-16 栏目: 出版日期: 2012-02-25
Title:
Rapid Identification of GmCRY1 and GmCRY2 Function by Leaf Injection in Soybean
文章编号:
1000-9841（2012）01-0013-04
作者:
秦正睿1; 2; 孙磊1; 2; 李宏宇2
1．吉林大学 植物科学学院，吉林 长春 130062；
2．中国农业科学院 作物科学研究所，北京 100081
Author(s):
QIN Zheng-rui1; 2; SUN Lei1; 2; LI Hong-yu2
1.College of Plant Science,Jilin University,Changchun 130062,Jilin；
2.Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 10081,China
关键词:
大豆; 隐花色素; 根癌农杆菌; 开花时间
Keywords:
Soybean; Cryptochrome genes; Agrobacterium tumefaciens; Flowering time
分类号:
S565.1
DOI:
10.3969/j.issn.1000-9841.2012.01.004
文献标志码:
A
摘要:
将大豆隐花色素基因GmCRY1、GmCRY2构建在N端含有绿色荧光蛋白(GFP)的pEGAD载体上，并转化农杆菌EHA105菌株。将含有上述基因的EHA105菌液分别注射到大豆单叶及复叶叶片中，观察长日条件下（16 h光照/8 h 黑暗）和短日条件下（8 h光照/16 h黑暗）注射植株的开花时间。结果叶片注射含GmCRY1基因菌液的植株在长日条件下早开花，通过RT-PCR检测表明注射叶片中GmCRY1成功表达，而叶片注射含GmCRY2基因菌液的植株虽然也通过RT-PCR检测到基因表达，但无论长日和短日条件下开花时间均无改变。结果证实注射叶片法可快速鉴定GmCRY1和GmCRY2基因对大豆开花时间的影响。
Abstract:
Soybean(Glycine max)Cryptochrome 1(GmCRY1)and Cryptochrome 2(GmCRY2)genes were constructed into expression vector pEGAD,and injected into soybean leaves by Agrobacterium tumefaciens EHA105 mediated transformation.The Agrobacterium tumefaciens EHA105 with vector pEGAD was used as control.The RT-PCR method was carried on to detect if the GmCRY1 and GmCRY2 were transient expressed in the leaves.The transformed plants were grown in different photoperiod light conditions(SD:8 h light/16 h darkness;LD:16 h light/ 8 h darkness)and the flowering data was calculated to conclude the effect of transient expression of GmCRY1 and GmCRY2 on the flowering time in soybean.The results indicated that the transient expressed GmCRY1 in soybean flowered earlier in LD,and the GmCRY1 expression was detected in the injected leaves through RT-PCR analysis.Although the expression of GmCRY2 was detected,it had no effect on flowering time in both LD and SD conditions.This result confirmed that the leaves injection method can quickly identify the function of GmCRY1 and GmCRY2 genes on soybean flowering time.

参考文献/References:

[1]Ritz T,Yoshii T,Helfrich-Foerster C,et al.Cryptochrome:a photoreceptor with the properties of a magneto receptor[J].Communicative Integrative Biology,2010,3(1):24-27.

[2]Kami?C,Lorrain S,Hornitschek P,et al.Light-regulated plant growth and development[J].Current Topics in Developmental Biology,2010,91:29-66.

[3]Zieker D,Jenne I,Koenigsrainer I,et al.Circadian expression of clock and tumor suppress or genes in human oral mucosa[J].Cellular Physiology and Biochemistry,2010,26(2):155-166.

[4]Cashmore A R.Cryptochromes:enabling plants and animals to determine circadian time[J].Cell,2003,114:537-543.

[5]杨志攀，周新安.大豆光周期遗传育种研究进展[J].中国油料作物学报，1999，21(1):67-73.(Yang Z P,Zhou X A.Soybean light cycle genetic breeding research progress[J].Chinese Journal of Oil Crop Sciences,1999,21(1):67-73.)

[6]陈洁敏,赵九洲,杨方人,等.播期对大豆开花及产量的影响[J].大豆科学，1998,17(3):225-229.(Chen J M,Zhao J Z,Yang F,et al.Effect of sowing dates on blooming model and grain yield of soybean[J].Soybean Science,1998,17(3):225-229.)

[7]党尉,卫志明.根癌农杆菌介导的高效大豆遗传转化体系的建立[J].分子细胞生物学报,2007,40(3):185-193.(Dang W,Wei Z M.Efficient Agrobacterium-mediated transformation of soybean[J].Journal of Molecular Cell Biology,2007,40(3):185-193.)

[8]李文霞，李文滨，吕文河，等.农杆菌介导大豆子叶节系统的两个问题突破[J].大豆科学,2008,27(1):173-180.(Li W X,Li W B,Lv W H,et al.Breakthrough of two questions on the?Agrobacterium-mediated soybean cotyledonary node systems[J].Soybean Science,2008,27(1):173-180.)

[9]马丽萍，胡正，张保缺，等.一种快速、高效的大豆农杆菌转化技术[J]中国农业科学，2008，41(3):661-668.(Ma L P,Hu Z,Zhang B Q, et al.A fast and high efficient technique for?Agrobacterium-Mediated transformation in soybean(Glycme max?L.Merrill)[J].Scientia Agricultura Sinica,2008,41(3):661-668.)

[10]徐香玲,李兴华,刘伟华,等.Ri质粒介导大豆花叶病毒外壳蛋白基因转化大豆的研究[J].大豆科学,1996,15(4):279-288.(Xu X L,Li X H,Liu W H,et al.Study on transferring soybeans mosaic vrious coat protein(SMV-CP)gene into soybean by Ri-plasmid[J].Soybean Science,1996,15(4):279-288.)

[11]王全伟,李新玲,张海玲,等.转Na+/H+逆向转运蛋白基因(nhaA)大豆植株的获得及耐盐性分析[J].分子植物育种,2010,8(4):701-707.(Wang Q W,Li X L,Zhang H L,et al.Overexpression of Na+?/H+antiporter gene(nhaA)improves salt tolerance in soybean[J].Molecular Plant Breeding,2010,8(4):701-707.)

[12]吕山花,樊颖伦,吕福堂,等.大豆GmPDS基因的克隆及VIGS表达载体构建和鉴定[J].生物技术通报,2010(4):122-131.(Lv S H,Fan Y L,Lv F T,et al.Construction and identification of VIGS expression vector carrying GmPDS gene cloned from Glycine max[J].Biotechnology Bulletin,2010(4):122-131.)

[13]王关林,方宏筠.植物基因工程[M].北京:科学出版社,2002.(Wang G L,Fang H J.Plant genetic engineering[M].Beijing:Science Press,2002.)

[14]皮伟，李名扬.根癌农杆菌介导FPF1基因转化菊花的研究[J].西南大学学报（自然科学版）,2007,29(4):70-73.(Pi W,Li M Y.Preliminary studies on transgenic chrysanthemum with?FPF1 gene mediated by Agrobactrium tumefaciens[J].Journal of Southwest University(Natural Science Edition),2007,29(4):70-73.)

[15]徐纪明,向太和.含GFP植物转基因表达载体的构建及在矮牵牛转基因不定根中的高效表达[J].遗传,2008,30(8):1069-1074.(Xu J M,Xiang T H.Construction of a novel vector harboring green fluorescence protein gene(GFP)and high expression of GFP?in transformed roots of Petunia hybrida[J].Hereditas,2008,30(8):1069-1074.)

[16]Lin C,Shalitin D.Cryptochrome structure and signal transduction[J].Annual Review of Plant Biology,2003,54:469-96.

[17]Zhang Q Z,Li H Y,Li R,et al.Association of the circadian rhythmic expression of GmCRY1a with a latitudinal cline in photoperiodic flowering of soybean[J].Proceedings of the National Academy Science,2008,105(52):21028-21033.

[18]Guo H,Yang H,Mockler T C,et al.Regulation of flowering time by Arabidopsis?photoreceptors[J].Science,1998,279:1360-1363.

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备注/Memo

基金项目：转基因专项资助项目（2009ZX08004-010B,2008ZX08010-004）;国家高技术研究发展计划资助项目（2006AA10Z107）。

第一作者简介：秦正睿（1985-），男，在读硕士，研究方向为植物分子生物学。E-mail：qzrv@163.com。
通讯作者：李宏宇（1970-），女，副研究员，博士，研究方向为植物分子生物学与基因工程。E-mail:lihongyu@caas.net。

更新日期/Last Update: 2014-08-14