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利用RNAi技术抑制马铃薯类受体激酶基因(StRLK)表达的研究

DOI: 10.3969/j.issn.1000-7091.2012.04.004, PP. 18-22

Keywords: 二倍体马铃薯,StRLK基因,遗传转化,RNA干扰

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Abstract:

植物类受体蛋白激酶(Receptor-likeproteinkinase,RLK)是蛋白激酶的一个亚家族,参与植物的信号转导,在植物生长发育、抗病等过程中起重要作用。在前期研究中自二倍体马铃薯高抗青枯病基因型ED13中获得了类受体蛋白激酶基因StRLK。利用StRLK基因的特异区段,以pUCCRNAi为中间克隆载体、pCHF1为植物表达载体,构建了该基因的RNA干扰载体pCHF1-StRLK。进一步以ED13的茎段为外植体,利用农杆菌介导法将pCHF1-StRLK导人ED13中,获得了5株再生植株。用CaMV35S启动子特异引物对5株再生植株进行PCR扩增,均得到大小约500bp的特异性条带,初步证明pCHF1-StRLK成功转入ED13。以ED13为对照,利用StRLK基因的特异引物对这5株再生植株进行半定量RT-PCR分析,结果表明,该基因的表达在5个转基因植株中均受到了不同程度的抑制,说明导入的pCHF1-StRLK发挥了干扰活性,且干扰效果与基因的插入位点有关,为进一步研究StRLK基因的功能提供依据。

References

[1]  《RNA干扰及其在植物中的研究进展》[J].草业科学,2011(5).
[2]  《水稻类受体激酶CRINKLY4胞外结合蛋白的研究》[J].湖北农业科学,2011(14).
[3]  Martin C,French E R. Bacterial wilt of potato [M]. Peru,li- ma:Technical Information Bulletin 13 CIP,1985; 16- 18.
[4]  Hawkes J G. The potato:evolution,biodiversity and genet- ic resources hard cover [M]. London: Belhaven Press, 1990.
[5]  Schwessinger B, Roux M, Kadota Y, et al. Phosphoryla- tion-dependent differential regulation of plant growth,cell death,and innate immunity by the regulatory receptor-like kinase BAKI[J]. PLoS Genet,2011,7 (4) : 1 - 17.
[6]  Li G C, Jin L P, Wang X W, et al. Gene transcription a- nalysis during interaction between potato and Ralstonia so- lanacearum [J]. Russian Journal of Plant Physiology. 2010,57(5) :685 -695.
[7]  Qu D Y. Use of unreduced gametes of potato (Solarium tuberosum L. ) for true potato seed production through 4x- 2x crosses [M]. Wageningen : Plant Breeding, 1996,104 : 38 - 46.
[8]  Hubbard S R, Miller W T. Receptor tyrosine kinases: mechanisms of activation and signaling [J]. Curr Opin CellBiol,2007,19(2) :117-123.
[9]  Zeng L, Velasquez A C, Munkvold K R, et al. A tomato LysM receptor-like kinase promotes immunity and its ki- nase activity is inhibited by AvrPtoB[J]. The Plant Jour- nal,2011,69(1) :92 - 103.
[10]  Rayapuram C,Jensen M K, Maiser F. Regulation of basal resistance by a powdery mildew-induced cysteine-rich receptor-like protein kinase in barley [J]. Mol Plant Pathol,2011,13(2) :135 - 147.
[11]  Papon N,Vansin A,Gantet P. Histidine-containing phos- photransfer domain extinction by RNA interference turns off a cytokinin signaling circuitry in Cathatanthus roseus suspension cells [J]. FEBS Lett,2004,558 : 85 - 88.
[12]  Koga A, Ishibashi T, Kimura S. Characterization of T- DNA insertion mutants and RNAi silenced plants of Ar- abidopsis thalianaUV-damaged DNA binding protein 2 (AtUV-DDB2) [J]. Plant Molecular Biology, 2006 (4) :227 -240.
[13]  Tremblay R,Diao H, Htiner N,et al. The development of a high-yield recombinant protein bioreactor throughRNAiin- duced knockdown of ATP/ADP transporter inSolanum tu- berosum[J]. J Biotechnol,2011,156( 1 ) :59 - 66.
[14]  Holeu T, Amarzguioui M, Wiigor M T, et al. Positional effects of short interfering RNAs targeting the human co- agulation trigger tissue factor [J]. Nucleic Acid Re- search, 2002,30 ( 8 ) : 1757 - 1766.
[15]  赵德尧.siRNA特异性及脱靶效应的研究[D].杭州:浙江理工大学,2010.
[16]  Jackson A L,Linsley P S. Recognizing and avoiding siR- NA off-target effects for target identification and thera- peutic application [J]. Nat Rev Drug Discov, 2010,9 (1) :57 -67.
[17]  《植物类受体蛋白激酶的研究进展》[J].河北师范大学学报:自然科学版,2010(2).
[18]  《RNAi在植物中的作用机理及其应用研究进展》[J].内蒙古民族大学学报:自然科学版,2010(1).

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