利用瞬时表达技术分析小麦抗白粉病相关基因CAF1、BI-1和DAD1的功能

doi:10.3969/j.issn.1000-7091.2012.05.006

华北农学报 ›› 2012, Vol. 27 ›› Issue (5): 28-32. doi: 10.3969/j.issn.1000-7091.2012.05.006

所属专题： 小麦；生物技术；

利用瞬时表达技术分析小麦抗白粉病相关基因CAF1、BI-1和DAD1的功能

王瑜晖, 辛明明, 宋娜, 彭福祥, 唐华山, 李映辉, 倪中福, 解超杰, 孙其信

中国农业大学农业生物技术国家重点实验室中国农业大学农业部作物基因组学与遗传改良重点开放实验室北京市作物遗传改良重点实验室教育部作物杂种优势研究与利用重点实验室中国农业大学农学与生物技术学院

收稿日期:2012-07-10 出版日期:2012-10-28
作者简介:王瑜晖(1986-)，女，广东广州人，在溪硕士，主要从事小麦抗病育种研究。
基金资助:
转基因专项(2011ZX08002-001);“973”项目(2009CB118300)

Analysis of the Functions of Three Wheat Powdery Mildew Resistance-related Genes by A Transient Expression

WANG Yu-hui, XIN Ming-ming, SONG Na, PENG Fu-xiang, TANG Hua-shan, LI Ying-hui, NI Zhong-fu, XIE Chao-jie, SUN Qi-xin

State Key Laboratory of AgroBiotechnology Key Laboratory of Crop Genomics and Genetic Improvement Ministry of Agriculture Beijing Key Laboratory of Crop Genetic Improvement Key Laboratory of Crop Heterosis Research & Utilization Ministry of Education College of Agronomy and Biotechnolog China Agricultural University Beijing 100193 China

Received:2012-07-10 Published:2012-10-28

摘要/Abstract

摘要： 利用小麦离体叶片瞬时表达技术分析了3个小麦抗白粉病相关基因CAF1、BI-1和DAD1的功能。根据小麦响应白粉菌侵染的差异表达基因芯片检测结果,筛选出表达差异的抗病相关基因CAF1和BI-1,并挑选与植物程序性死亡相关的基因DAD1,将克隆到的这3个小麦基因全长ORF序列构建入超表达载体pAHC25中,利用瞬时表达技术检测目标基因和GUS基因共转化的阳性小麦叶片表皮细胞中白粉菌成功侵入并形成吸器的细胞比例(侵入频率),研究目标基因的超表达后对白粉菌入侵及吸器形成产生的影响。结果表明,小麦CAF1基因在感病小麦品种叶片中的瞬时表达,对白粉病侵入和吸器形成具有一定的抑制作用(侵入频率为(37.37±3.2)%,对照为(54.75±0.6)%),在一定程度上增强了表达细胞对白粉菌的抗性;表达DAD1基因则可能在一定程度上有利于白粉菌吸器的形成(侵入频率为(57.34±1.1)%,对照为(50.93±1.3)%);而在感病叶片中表达基因BI-1对白粉菌吸器形成没有明显影响(侵入频率为(54.31±1.7)%,对照为(52.37±1.8)%)。

关键词: 小麦白粉菌, 瞬时表达技术, CAF1基因, BI-1基因, DAD1基因

Abstract: Transient expression system was used to analyze the function of three wheat powdery mildew resistance-related genes,CAF1,BI-1,and DAD1.Target genes were constructed into vectors pAHC25 and transformed into leaf epidermal cells of a powdery mildew-susceptible wheat variety Jingdong 8 by bombardment.GUS gene was co-transformed with target genes to mark the transformed cells.The penetration efficiency(PE) after bombarding of target gene was compared with control,then evaluated the effects of the target gene′s products on the invasion of powdery mildew.The results implied that transient expression of gene CAF1 could partly inhibit the penetration of conidia spores and formation of haustoria(PE was reduced to(37.37±3.2)%,as compared with the control of(54.75±0.6)%),while gene BI-1(PE was(54.31±1.7)%,PE of control was(52.37±1.8)%) and gene DAD1(PE was(57.34±1.1)%,PE of control was(50.93±1.3)%)showed no inhibiting effects.

Key words: Wheat powdery mildew, Transient expression, CAF1 BI-1 DAD1

中图分类号:

S435.121.3

王瑜晖, 辛明明, 宋娜, 彭福祥, 唐华山, 李映辉, 倪中福, 解超杰, 孙其信. 利用瞬时表达技术分析小麦抗白粉病相关基因CAF1、BI-1和DAD1的功能[J]. 华北农学报, 2012, 27(5): 28-32. doi: 10.3969/j.issn.1000-7091.2012.05.006.

WANG Yu-hui, XIN Ming-ming, SONG Na, PENG Fu-xiang, TANG Hua-shan, LI Ying-hui, NI Zhong-fu, XIE Chao-jie, SUN Qi-xin. Analysis of the Functions of Three Wheat Powdery Mildew Resistance-related Genes by A Transient Expression[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(5): 28-32. doi: 10.3969/j.issn.1000-7091.2012.05.006.

http://www.hbnxb.net/CN/Y2012/V27/I5/28

参考文献

[1] Schweizer P,Pokorny J,Abderhalden O. A transient assay system for the functional assessment of defense-related genes in wheat[J]. Mol Plant Microbe Interact, 1999,12 (8) :647 -654.

[2] Nielsen K, Olsen O. A transient expression system to assay putative antifungal genes on powdery mildew infected barley leaves[J]. Physiol Mol Plant Pathol, 1999,54 : 1 12.

[3] Nelson A J,Bushnell W R. Transient expression of anthocyanin genes in barley epidermal cells:potential for use in evaluation of disease response genes [J]. Transgenic Res, 1997,6:233 244.

[4] 辛明明.小麦Pr030近等基因系响应白粉菌侵染的转录组表达谱与小RNA表达谱分析[D].北京:中国农业大学作物遗传育种系,2011.

[5] Bogdan J A, Adams-burton C, Pedicord D L,et al. Human carbon catabolite repressor protein (CCR4)-associative factor 1 :cloning,expression and characterization of its interaction with the B-cell translocation protein BTG1 [J]. Biochem J, 1998,336:471 481.

[6] Sarowar S, Oh H W, Cho H S, et al. Capsicum annuum CCR4-associated factor CaCAF1 is necessary for plant development and defense response [J]. The Plant Journal, 2007,51:792 802.

[7] Liang W X, Li C B, Liu F, et al. The Arabidopsis homologs of CCRg-associated factor 1 show mRNA deadenylation activity and play a role in plant defense responses [J]. Cell Research ,2009,19:307 316.

[8] Walley J W, Kelley D R, Savchenko T,et al. Investigating the function of CAF1 deadenylases during plant stress responses [J]. Plant Signaling & Behavior, 2010, 5 ( 7 ) : 802 805.

[9] Chae H J, Ke N, Kim H R,et al. Evolutionarily conserved cytoprotection provided by Bax inhibitor-1 homologs from animals, plants, and yeast [J]. Gene, 2003,323 : 101 113.

[10] Watanabe N, Eric L. Bax inhibitor-1, a conserved cell death suppressor,is a key molecular switch downstream from a variety of biotic and abiotic stress signals in plants[J]. Int J Mol Sci ,2009,10:3149 3167.

[11] Raft M C. Social control on cell survival and cell death [J]. Nature, 1992,356:397 400.

[12] Schweizer P,Christoffel A. Transient expression of members of the germin-like gene family in epidermal ceils of wheat confers disease resistance [J]. Plant J, 1999,20 (5) :540 -552.

[13] Kawai M, Pan L, Reed J C, et al. Evolutionally conserved plant homologue of the Bax Inhibitor-1 (BI-1) gene capable of suppressing Bax-induced cell death in yeast [J]. FEBS Lett, 1999,46.4 : 143 147.

[14] Eichmann R, Bischof M, Weis C, et al. BAX INHIBITOR-1 is required for full susceptibility of barley to powdery mildew[J]. Mol Plant Microbe Interact,2010, 23(9) :1217 1227.

[15] Babaeizad V,lmani J, Kogel K H, et al. Over-expression of the cell death regulator BAX inhibitor-1 in barley confers reduced or enhanced susceptibility to distinct fungal pathogens [J]. Theor Appl Genet, 2009,118 ( 3 ) : 455 465.

[16] Eichmann R,H ckelhoven R. Accommodation of powdery mildew fungi in intact plant ceils [J]. J Plant Physiol, 2008,165:5 18.

[17] Eichmann R, Dechert C, Kogel K H, et al. Transient over-expression of barley BAX Inhibitor-1 weakens oxidative defence and MLA12-mediated resistance to Blumeria graminis f. sp. hordei [J]. Mol Plant Pathol, 2006,7 (6) -543 -552.

[18] Eichmann R, Schultheiss H, Kogel K-H,et al. The barley apoptosis suppressor homologue BAX inhibitor-1 compromises nonhost penetration resistance of barley to the inappropriate pathogen Blumeria graminis f. sp. Tritici [J]. Molecular Plant Microbe Interactions, 2004, 17 (5) :484 -490.

[19] Gallois P, Makishima T, Hechtt V, et al. An Arabidopss thaliana cDNA complementing a hamster apoptosis suppressor mutant [J]. Plant Journal, 1997, 11 : 1325 1331.

[20] Lindholm P, Kuittinen T,Sorri O, et al. Glycosylation of phytepsin and expression of dadl, dad2 and ostl during onset of cell death in germinating barley scutella [J]. Mech Dev ,2000,93 ( 1/2 ) : 169 73.

[21] Apte S S,Mattei M G,Seldin M F,et al. The highly conserved defender against the death (DAD1) gene maps to human chromosome 14q11-q12 and mouse chromosome 14 and has plant and nematode homologs [J]. FEBS Lett, 1995,363:304 306.

[22] 龚文芳.棉花抗细胞凋亡基因GhDAD1的克隆、定位及表达分析[D].武汉:华中农业大学作物遗传育种系,2010.

[23] 孙晓丽.小麦抗白粉病基因的分子标记及白粉菌引发的细胞程序性死亡的相关研究[D].北京:中国农业科学院,2006.

[24] 《荧光素钠和考马斯亮蓝应用于小麦白粉病菌染色效果比较》[J].菌物系统,2002(4).

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利用瞬时表达技术分析小麦抗白粉病相关基因CAF1、BI-1和DAD1的功能

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[1]	林晓民, 侯文邦, 周丽鸿, 商鸿生. 小麦白粉菌寄主范围研究初报[J]. 华北农学报, 1995, 10(1): 82-87.
[2]	王锡锋, 何家泌. 小麦品种(系)与小麦白粉菌的相互作用Ⅰ.小麦白粉菌毒力频率和毒性基因的研究[J]. 华北农学报, 1988, 3(1): 73-79.

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