全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

快速筛选拟南芥受精和早期胚胎发生相关基因的方法

DOI: 10.11913/PSJ.2095-0837.2015.40564, PP. 564-571

Keywords: 拟南芥,T-DNA,受精,早期胚胎发生,突变体筛选

Full-Text   Cite this paper   Add to My Lib

Abstract:

?阐明拟南芥受精和早期胚胎发生过程对理解被子植物生殖发育有着重要的指导意义,而利用正向遗传学方法研究拟南芥突变体的表型及其分子机理是探究植物基因功能最常用的一种方法。基于常规的插入突变(包括T-DNA和转座子)、化学诱变(如ethylmethanesulfonate,EMS)和高能射线方法构建的突变体库中假阳性突变体多,难以高效筛选到受精和早期胚胎发生相关基因的突变体。为解决这一难题,本研究建立了一种构建T-DNA插入突变体文库的新方法。即在载体pCAMBIA1302的T-DNA元件上增加花粉特异荧光标记基因(pLAT52::EGFP),并遗传转化具有四分体花粉的Columbia野生型拟南芥突变体qrt1-2;对获得的突变体库可利用花粉荧光快速排除假阳性突变体,并采用反向PCR(inverse-PCR)扩增技术确定突变位点。此方法在筛选拟南芥受精和早期胚胎发生相关基因突变体上的成功应用表明,其是一种效率高、针对性强、操作相对快捷方便的拟南芥突变体筛选方法。

References

[1]  曹冬梅,范喜英,王云山,唐黎芳. 拟南芥激活标签突变体库的构建及突变体表型的分析[J] .农业生物技术学报,2008,16(2): 292-298.
[2]  The Arabidopsis Genome Initiative. Analysis of the genome sequence of the flowering plant Arabidopsis thaliana[J]. Nature,2000, 408(6814): 796-815.
[3]  孙万刚. 拟南芥突变体构建方法及目的基因分离鉴定技术[J]. 安徽农业科学,2010,38(18): 9433- 9434.
[4]  Greene EA, Codomo CA, Taylor NE, Henikoff JG, Till BJ, Reynolds SH, Enns LC, Burtner C, Johnson JE, Odden AR, Comai L, Henikoff S. Spectrum of chemically induced mutations from a large-scale reverse-genetic screen in Arabidopsis[J]. Genetics, 2003, 164(2): 731-740.
[5]  Parry MA, Madgwick PJ, Bayon C, Tearall K, Hernandez-Lopez A, Baudo M, Rakszegi M, Hamada W, Al-Yassin A, Ouabbou H, Labhilili M,Phillips AL. Mutation discovery for crop improvement[J]. J Exp Bot, 2009, 60(10): 2817-2825.
[6]  宋磊, 杨仲南, 吴世福, 崔永兰. 拟南芥网状突变体E-210基因精细定位[J].植物科学学报,2008, 26(5): 437-442.
[7]  Springer PS. Gene traps: tools for plant development and genomics[J]. Plant Cell, 2000, 12(7): 1007-1020.
[8]  An G, Lee S, Kim SH, Kim SR. Molecular gene-tics using T-DNA in rice[J]. Plant Cell Physiol, 2005, 46(1): 14-22.
[9]  Schneeberger RG, Zhang K, Tatarinova T, Troukhan M, Kwok SF, Drais J, Klinger K, Orejudos F, Macy K, Bhakta A, Burns J, Subramanian G, Donson J, Flavell R, Feldmann KA. Agrobacterium T-DNA integration in Arabidopsis is correlated with DNA sequence compositions that occur frequently in gene promoter regions[J]. Funct Integr Genomics, 2005, 5(4): 240-253.
[10]  崔清志,刘晓红,陈惠明. EMS诱变技术研究进展[J]. 湖南农业科学,2013,13(5): 7-9.
[11]  阎双勇,谭振波,李仕贵. 水稻插入突变体库构建研究进展[J]. 中国生物工程杂志,2004,24(6): 48-53.
[12]  Brink RA, Cooper DC. Double fertilization and development of the seed in angiosperms[J]. Botanical Gazette, 1940, 102(1): 1-25.
[13]  Preuss D, Rhee SY, Davis RW. Tetrad analysis possible in Arabidopsis with mutation of the QUARTET(QRT)genes[J]. Science, 1994, 264(5164): 1458-1460.
[14]  Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures[J]. Physiologia Plantarum, 1962, 15(3): 473-497.
[15]  Clough SJ,Bent AF. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana[J]. Plant J, 1998, 16(6): 735-743.
[16]  曾凡锁,詹亚光. 转基因植物中外源基因的整合特性及其研究策略[J]. 植物学通报,2004,21(5): 565-577.
[17]  梁春莉,刘孟军,赵锦. 植物种子败育研究进展[J]. 分子植物育种,2005,3(1): 117-122.
[18]  Mathur J, Szabados L, Schaefer S, Grunenberg B, Lossow A, Jonas-Straube E, Schell J, Koncz C,Koncz-Kalman Z. Gene identification with sequenced T-DNA tags generated by transformation of Arabidopsis cell suspension[J]. Plant J, 1998, 13(5): 707-716.
[19]  Souer E, Quattrocchio F, de Vetten N, Mol J, Koes R. A general method to isolate genes tagged by a high copy number transposable element[J]. Plant J, 1995, 7(4): 677-685
[20]  赵霞,周波,李玉花. T-DNA 插入突变在植物功能基因组学中的应用[J]. 生物技术通讯,2009,6(20): 880-885.
[21]  Chen YH, Li HJ, Shi DQ, Yuan L, Liu J, Sreeni-vasan R, Baskar R, Grossniklaus U, Yang WC. The central cell plays a critical role in pollen tube guidance in Arabidopsis[J]. Plant Cell, 2007, 19(11): 3563-3577.
[22]  Gross-Hardt R, Kagi C, Baumann N, Moore JM, Baskar R, Gagliano WB, Jurgens G, Grossniklaus U. LACHESIS restricts gametic cell fate in the female gametophyte of Arabidopsis[J]. PLoS Biol, 2007, 5(3): 494-500.
[23]  Wu JJ, Peng XB, Li WW, He R, Xin HP, Sun MX. Mitochondrial GCD1 dysfunction reveals reciprocal cell-to-cell signaling during the maturation of Arabidopsis female gametes[J]. Dev Cell, 2012, 23(5): 1043-1058.

Full-Text

Contact Us

[email protected]

QQ:3279437679

WhatsApp +8615387084133