云南疣粒野生稻稻瘟病抗性
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摘要
野生稻(Oryza rufipogo)保存有许多栽培稻(O.sativa)不具备或已经消失的优异基因资源,是扩大栽培稻遗传背景、改良产量与品质、提高抗病虫害及抗逆境能力的重要基因库。疣粒野生稻(O.meyeriana)是中国3种野生稻资源之一,主要分布在云南。为进一步了解其稻瘟病抗性,首先利用来自不同稻作区的稻瘟病菌株,通过注射接种法对疣粒野生稻进行系统的稻瘟病抗性鉴定,发现疣粒野生稻对接种的所有稻瘟病菌株都感病。进一步采用3'/5'RACE方法,从疣粒野生稻中克隆了水稻同源基因Pid2和Pid3,并构建过表达转基因株系对基因功能进行了研究。结果表明,Pid2和Pid3与疣粒野生稻中同源基因间在DNA和氨基酸水平上有较大的序列差异,过表达转基因的日本晴植株对稻瘟病菌的敏感性与对照相似。推测疣粒野生稻在自然接种条件下,表现出的抗稻瘟病表型很可能是其旱生叶片结构特征形成了对稻瘟病菌侵染的天然屏障。对控制疣粒野生稻这一类性状基因资源的挖掘和利用,有利于优良抗性水稻品种的培育。研究结果为疣粒野生稻的研究利用提供了新信息和新思路。
Wild species of Oryza contain numerous genes of economic importance and are being used as an important cultivated rice gene pool to expand the genetic background, improve yield and quality, and increase the ability for resistance or tolerance to pests as well as bioic and abiotic stresses. O. meyeriana is one of three native wild rice species in China, mainly distributed in Yunnan province. To further understand the disease resistance to rice blast, we performed a systematic investigation of rice blast resistance with O. meyeriana. O. meyeriana plants were inoculated by injection with multiple blast strains from different rice planting areas. Then, we cloned and functionally identified Pid2 and Pid3 orthologous genes in O. meyeriana. O. meyeriana was sensitive to all blast isolates used in this experiment. The DNA and amino acid sequences significantly differed between Pid2 and its orthologs in O. meyeriana, as did Pid3 and its orthologs. Nipponbare plants overexpressing the orthologous genes of Pid2 and Pid3 showed mildly enhanced susceptibility to Magnaporthe oryzae. The orthologs of blast-resistant genes in O. meyeriana may be susceptible alleles. O. meyeriana exhibited resistance to rice blast under natural inoculation likely due to a xeromorphic leaf structure that reduces susceptibility to M. oryzae. Characterization of these genes will help breed elite rice. Our study provides new information and ideas for research and utilization of O. meyeriana.
Wild species of Oryza contain numerous genes of economic importance and are being used as an important cultivated rice gene pool to expand the genetic background, improve yield and quality, and increase the ability for resistance or tolerance to pests as well as bioic and abiotic stresses. O. meyeriana is one of three native wild rice species in China, mainly distributed in Yunnan province. To further understand the disease resistance to rice blast, we performed a systematic investigation of rice blast resistance with O. meyeriana. O. meyeriana plants were inoculated by injection with multiple blast strains from different rice planting areas. Then, we cloned and functionally identified Pid2 and Pid3 orthologous genes in O. meyeriana. O. meyeriana was sensitive to all blast isolates used in this experiment. The DNA and amino acid sequences significantly differed between Pid2 and its orthologs in O. meyeriana, as did Pid3 and its orthologs. Nipponbare plants overexpressing the orthologous genes of Pid2 and Pid3 showed mildly enhanced susceptibility to Magnaporthe oryzae. The orthologs of blast-resistant genes in O. meyeriana may be susceptible alleles. O. meyeriana exhibited resistance to rice blast under natural inoculation likely due to a xeromorphic leaf structure that reduces susceptibility to M. oryzae. Characterization of these genes will help breed elite rice. Our study provides new information and ideas for research and utilization of O. meyeriana.
引文
胡朝芹,刘剑宇,王韵茜,杨睿,汪秉琨,何月秋,曾千春,罗琼(2017).粳稻子预44抗LP11稻瘟病菌基因Pizy6(t)的定位.植物学报52,61-69.
蒋春苗,黄兴奇,李定琴,余腾琼,程在全(2012).云南野生稻叶茎根组织结构特性的比较研究.西北植物学报32,99-105.
李冬波,张端品,林兴华(2009).SSR标记在疣粒野生稻和普通栽培稻中的多态性研究.华中农业大学学报28,1-4.
梁斌,肖放华,黄费元,彭绍裘,陈勇,戴陆国,刘二明(1999).云南野生稻对稻瘟病的抗性评价.中国水稻科学13,183-185.
刘继梅,程在全,杨明挚,吴成军,王玲仙,孙一丁,黄兴奇(2003).云南3种野生稻中抗病基因同源序列的克隆及序列分析.中国农业科学36,273-280.
刘进,刘建锋,曾义(2001).寄主植物的形态结构抗病性.四川林业科技22(3),54-56.
彭绍裘,魏子生,毛昌祥,黄河清,肖放华,罗宽(1982).云南省疣粒野生稻、药用野生稻和普通野生稻多抗性鉴定.植物病理学报12,60-62,74.
盛腊红,何光存,舒理慧,廖兰杰(1999).提高疣粒野生稻愈伤组织分化能力的研究.植物学通报16,614-617.
文婷,梁毅,江南,李智强,金灵,杨婷婷,戴良英,王国梁,刘雄伦(2012).利用pi9基因序列标记辅助选择改良籼稻稻瘟病抗性.湖南农业大学学报(自然科学版)38,262-266.
颜惠霞,徐秉良,梁巧兰,薛应钰,陈荣贤,梁志福(2009).南瓜品种对白粉病的抗病性与叶绿素含量和气孔密度的相关性.植物保护35,79-81.
杨健源,康金平,黄显良,姜先芽,赖添奎,陈深,李传瑛,曾先列,朱小源(2007).稻瘟病田间自然诱发病圃的建立及其在抗性鉴定中的应用.广东农业科学(9),59-61.
云勇,韩义胜(2014).我国野生稻资源的抗病性鉴定与利用研究进展.植物遗传资源学报15,472-476,482.
张乃群,杨莉萍,杜敏华(1999).我国三种野生稻及两个栽培稻品种叶片表面亚显微结构的观察研究.南都学坛(自然科学版)19(3),70-73.
周镕,王波,杨睿,李书,樊琳琳,曾千春,罗琼(2015).粳稻子预44中稻瘟病数量抗性位点分析.植物学报50,691-698.
Chen XW,Shang JJ,Chen DX,Lei CL,Zou Y,Zhai WX,Liu GZ,Xu JC,Ling ZZ,Cao G,Ma BT,Wang YP,Zhao XF,Li SG,Zhu LH(2006).A B-lectin receptor kinase gene conferring rice blast resistance.Plant J 46,794-804.
Das A,Soubam D,Singh PK,Thakur S,Singh NK,Sharma TR(2012).A novel blast resistance gene,Pi54rh cloned from wild species of rice,Oryza rhizomatis confers broad spectrum resistance to Magnaporthe oryzae.Funct Integr Genomics 12,215-228.
Fu CY,Wu T,Liu WG,Wang F,Li JH,Zhu XY,Huang HJ,Liu ZR,Liao YL,Zhu MS,Chen JW,Huang YJ(2012).Genetic improvement of resistance to blast and bacterial blight of the elite maintainer line Rongfeng B in hybrid rice(Oryza sativa L.)by using marker-assisted selection.Afr JBiotechnol 11,13104-13124.
Jena KK(2010).The species of the genus Oryza and transfer of useful genes from wild species into cultivated rice,O.sativa.Breed Sci 60,518-523.
Jeung JU,Kim BR,Cho YC,Han SS,Moon HP,Lee YT,Jena KK(2007).A novel gene,Pi40(t),linked to the DNAmarkers derived from NBS-LRR motifs confers broad spectrum of blast resistance in rice.Theor Appl Genet115,1163-1177.
Koide Y,Kawasaki A,Telebanco-Yanoria MJ,Hairmansis A,Nguyet NTM,Bigirimana J,Fujita D,Kobayashi N,Fukuta Y(2010).Development of pyramided lines with two resistance genes,Pish and Pib,for blast disease(Magnaporthe oryzae B.Couch)in rice(Oryza sativa L.).Plant Breed 129,670-675.
Liu WD,Liu JL,Triplett L,Leach JE,Wang GL(2013).Novel insights into rice innate immunity against bacterial and fungal pathogens.Annu Rev Phytopathol 52,213-241.
Lv QM,Xu X,Shang JJ,Jiang GH,Pang ZQ,Zhou ZZ,Wang J,Liu Y,Li T,Li XB,Xu JC,Cheng ZK,Zhao XF,Li SG,Zhu LH(2013).Functional analysis of Pid3-A4,an ortholog of rice blast resistance gene Pid3 revealed by allele mining in common wild rice.Phytopathology 103,594-599.
Nishimura A,Ashikari M,Lin SY,Takashi T,Angeles ER,Yamamoto T,Matsuoka M(2005).Isolation of a rice regeneration quantitative trait loci gene and its application to transformation systems.Proc Natl Acad Sci USA 102,11940-11944.
Shang JJ,Yong T,Chen XW,Zou Y,Lei CL,Wang J,Li XB,Zhao XF,Zhang MJ,Lu ZK,Xu JC,Cheng ZK,Wan JM,Zhu LH(2009).Identification of a new rice blast resistance gene,Pid3,by genomewide comparison of paired nucleotide-binding site-leucine-rich repeat genes and their pseudogene alleles between the two sequenced rice genomes.Genetics 182,1303-1311.
Skamnioti P,Gurr SJ(2009).Against the grain:safeguarding rice from rice blast disease.Trends Biotechnol27,141-150.
Standard Evaluation System(SES)for Rice(2013).Authority Files for International Rice Research Institute,5th edition.http://www.clrri.org/ver2/uploads/SES.
Tanweer FA,Rafii MY,Sijam K,Rahim HA,Ahmed F,Latif MA(2015).Current advance methods for the identification of blast resistance genes in rice.CR Biol 338,321-334.
Tharreau D,Lebrun MH,Talbot NJ,Notteghem JL(2000).Advances in Rice Blast Research.Netherlands:Springer Press.pp.54-62.
Vaughan DA(1994).The Wild Relatives of Rice.A Genetic Resources Handbook.Manila:International Rice Research Institute.pp.457-462.
Xiao N,Wu YY,Pan CH,Yu L,Chen Y,Liu GQ,Li YH,Zhang XX,Wang ZP,Dai ZY,Liang CZ,Li AH(2016).Improving of rice blast resistances in Japonica by pyramiding major R genes.Front Plant Sci 7,1918.
Zou XH,Zhang FM,Zhang JG,Zhang LL,Tang L,Wang J,Sang T,Ge S(2008).Analysis of 142 genes resolves the rapid diversification of the rice genus.Genome Biol 9,R49.
蒋春苗,黄兴奇,李定琴,余腾琼,程在全(2012).云南野生稻叶茎根组织结构特性的比较研究.西北植物学报32,99-105.
李冬波,张端品,林兴华(2009).SSR标记在疣粒野生稻和普通栽培稻中的多态性研究.华中农业大学学报28,1-4.
梁斌,肖放华,黄费元,彭绍裘,陈勇,戴陆国,刘二明(1999).云南野生稻对稻瘟病的抗性评价.中国水稻科学13,183-185.
刘继梅,程在全,杨明挚,吴成军,王玲仙,孙一丁,黄兴奇(2003).云南3种野生稻中抗病基因同源序列的克隆及序列分析.中国农业科学36,273-280.
刘进,刘建锋,曾义(2001).寄主植物的形态结构抗病性.四川林业科技22(3),54-56.
彭绍裘,魏子生,毛昌祥,黄河清,肖放华,罗宽(1982).云南省疣粒野生稻、药用野生稻和普通野生稻多抗性鉴定.植物病理学报12,60-62,74.
盛腊红,何光存,舒理慧,廖兰杰(1999).提高疣粒野生稻愈伤组织分化能力的研究.植物学通报16,614-617.
文婷,梁毅,江南,李智强,金灵,杨婷婷,戴良英,王国梁,刘雄伦(2012).利用pi9基因序列标记辅助选择改良籼稻稻瘟病抗性.湖南农业大学学报(自然科学版)38,262-266.
颜惠霞,徐秉良,梁巧兰,薛应钰,陈荣贤,梁志福(2009).南瓜品种对白粉病的抗病性与叶绿素含量和气孔密度的相关性.植物保护35,79-81.
杨健源,康金平,黄显良,姜先芽,赖添奎,陈深,李传瑛,曾先列,朱小源(2007).稻瘟病田间自然诱发病圃的建立及其在抗性鉴定中的应用.广东农业科学(9),59-61.
云勇,韩义胜(2014).我国野生稻资源的抗病性鉴定与利用研究进展.植物遗传资源学报15,472-476,482.
张乃群,杨莉萍,杜敏华(1999).我国三种野生稻及两个栽培稻品种叶片表面亚显微结构的观察研究.南都学坛(自然科学版)19(3),70-73.
周镕,王波,杨睿,李书,樊琳琳,曾千春,罗琼(2015).粳稻子预44中稻瘟病数量抗性位点分析.植物学报50,691-698.
Chen XW,Shang JJ,Chen DX,Lei CL,Zou Y,Zhai WX,Liu GZ,Xu JC,Ling ZZ,Cao G,Ma BT,Wang YP,Zhao XF,Li SG,Zhu LH(2006).A B-lectin receptor kinase gene conferring rice blast resistance.Plant J 46,794-804.
Das A,Soubam D,Singh PK,Thakur S,Singh NK,Sharma TR(2012).A novel blast resistance gene,Pi54rh cloned from wild species of rice,Oryza rhizomatis confers broad spectrum resistance to Magnaporthe oryzae.Funct Integr Genomics 12,215-228.
Fu CY,Wu T,Liu WG,Wang F,Li JH,Zhu XY,Huang HJ,Liu ZR,Liao YL,Zhu MS,Chen JW,Huang YJ(2012).Genetic improvement of resistance to blast and bacterial blight of the elite maintainer line Rongfeng B in hybrid rice(Oryza sativa L.)by using marker-assisted selection.Afr JBiotechnol 11,13104-13124.
Jena KK(2010).The species of the genus Oryza and transfer of useful genes from wild species into cultivated rice,O.sativa.Breed Sci 60,518-523.
Jeung JU,Kim BR,Cho YC,Han SS,Moon HP,Lee YT,Jena KK(2007).A novel gene,Pi40(t),linked to the DNAmarkers derived from NBS-LRR motifs confers broad spectrum of blast resistance in rice.Theor Appl Genet115,1163-1177.
Koide Y,Kawasaki A,Telebanco-Yanoria MJ,Hairmansis A,Nguyet NTM,Bigirimana J,Fujita D,Kobayashi N,Fukuta Y(2010).Development of pyramided lines with two resistance genes,Pish and Pib,for blast disease(Magnaporthe oryzae B.Couch)in rice(Oryza sativa L.).Plant Breed 129,670-675.
Liu WD,Liu JL,Triplett L,Leach JE,Wang GL(2013).Novel insights into rice innate immunity against bacterial and fungal pathogens.Annu Rev Phytopathol 52,213-241.
Lv QM,Xu X,Shang JJ,Jiang GH,Pang ZQ,Zhou ZZ,Wang J,Liu Y,Li T,Li XB,Xu JC,Cheng ZK,Zhao XF,Li SG,Zhu LH(2013).Functional analysis of Pid3-A4,an ortholog of rice blast resistance gene Pid3 revealed by allele mining in common wild rice.Phytopathology 103,594-599.
Nishimura A,Ashikari M,Lin SY,Takashi T,Angeles ER,Yamamoto T,Matsuoka M(2005).Isolation of a rice regeneration quantitative trait loci gene and its application to transformation systems.Proc Natl Acad Sci USA 102,11940-11944.
Shang JJ,Yong T,Chen XW,Zou Y,Lei CL,Wang J,Li XB,Zhao XF,Zhang MJ,Lu ZK,Xu JC,Cheng ZK,Wan JM,Zhu LH(2009).Identification of a new rice blast resistance gene,Pid3,by genomewide comparison of paired nucleotide-binding site-leucine-rich repeat genes and their pseudogene alleles between the two sequenced rice genomes.Genetics 182,1303-1311.
Skamnioti P,Gurr SJ(2009).Against the grain:safeguarding rice from rice blast disease.Trends Biotechnol27,141-150.
Standard Evaluation System(SES)for Rice(2013).Authority Files for International Rice Research Institute,5th edition.http://www.clrri.org/ver2/uploads/SES.
Tanweer FA,Rafii MY,Sijam K,Rahim HA,Ahmed F,Latif MA(2015).Current advance methods for the identification of blast resistance genes in rice.CR Biol 338,321-334.
Tharreau D,Lebrun MH,Talbot NJ,Notteghem JL(2000).Advances in Rice Blast Research.Netherlands:Springer Press.pp.54-62.
Vaughan DA(1994).The Wild Relatives of Rice.A Genetic Resources Handbook.Manila:International Rice Research Institute.pp.457-462.
Xiao N,Wu YY,Pan CH,Yu L,Chen Y,Liu GQ,Li YH,Zhang XX,Wang ZP,Dai ZY,Liang CZ,Li AH(2016).Improving of rice blast resistances in Japonica by pyramiding major R genes.Front Plant Sci 7,1918.
Zou XH,Zhang FM,Zhang JG,Zhang LL,Tang L,Wang J,Sang T,Ge S(2008).Analysis of 142 genes resolves the rapid diversification of the rice genus.Genome Biol 9,R49.