江苏省稻瘟病菌致病力分化及无毒基因组成分析
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摘要
针对2013-2017年从江苏省采集标样并分离得到的共计621个稻瘟病菌[Magnaporthe oryzae (Hebert)Barr]单孢菌株,利用10对稻瘟病菌无毒基因(ACE1、PWL1、Avr-Pita、Avr1-CO39、Avr-Pizt、Avr-Pib、Avr-Pia、Avr-Pii、Avr-Pik、Avr-Pi9)特异性引物进行扩增,分析江苏省稻瘟病菌无毒基因的分布及频率。结果表明:上述10个无毒基因在江苏省均有分布,但出现频率不同,其中平均扩增频率最高的为Avr-Pib,达到74. 56%,最低的是Avr-Pia,只有9. 34%。Avr-Pib、Avr-Pik、Avr-Pizt连续5年扩增频率较高且稳定遗传。此外,采用日本清泽鉴别品种对稻瘟病菌供试菌株进行致病力测定,结果表明供试菌株对Pi-zt、Pi-z、Pi-ta、Pi-ta2、Pi-i表现为弱毒力。综上所述,江苏省稻瘟病菌群体组成较为复杂,且存在较大易变性;无毒基因的分布存在较大差异,且与地理区域密切相关。
In this study,10 pairs of specific primers of avirulence genes( ACE1,PWL1,AvrP i-ta,Avr1-CO39,AvrPizt,Avr-Pib,Avr-Pia,Avr-Pii,Avr-Pik and Avr-Pi9) in a total of 621 single spore strains of Magnaporthe oryzae [Magnaporthe oryzae( Hebert) Barr] isolated from Jiangsu province during 2013-2017 were used to analyze the distribution and frequency of avirulence genes. The PCR results showed that the above 10 avirulence genes were distributed in M. oryzae strains of Jiangsu province,but the frequency of occurrence was different. Among them,Avr-Pib and Avr-Pia showed the highest and lowest frequency( 74.56% and 9.34%,respectively),Avr-Pib,Avr-Pik and Avr-Pizt had high frequency of amplification and stable heritability for five consecutive years. In addition,the pathogenicity of the M. oryzae was evaluated by using Kiyosawa' s differential varieties( KDV). The results revealed that the tested stains showed weak virulence against Pizt,Pi-z,Pi-ta,Pi-ta2,Pi-i. In summary,the population composition of M. oryzae in Jiangsu province is relatively complex,and it comes instable. The distribution of avr-genes is quite different and closely related to the geographical area.
In this study,10 pairs of specific primers of avirulence genes( ACE1,PWL1,AvrP i-ta,Avr1-CO39,AvrPizt,Avr-Pib,Avr-Pia,Avr-Pii,Avr-Pik and Avr-Pi9) in a total of 621 single spore strains of Magnaporthe oryzae [Magnaporthe oryzae( Hebert) Barr] isolated from Jiangsu province during 2013-2017 were used to analyze the distribution and frequency of avirulence genes. The PCR results showed that the above 10 avirulence genes were distributed in M. oryzae strains of Jiangsu province,but the frequency of occurrence was different. Among them,Avr-Pib and Avr-Pia showed the highest and lowest frequency( 74.56% and 9.34%,respectively),Avr-Pib,Avr-Pik and Avr-Pizt had high frequency of amplification and stable heritability for five consecutive years. In addition,the pathogenicity of the M. oryzae was evaluated by using Kiyosawa' s differential varieties( KDV). The results revealed that the tested stains showed weak virulence against Pizt,Pi-z,Pi-ta,Pi-ta2,Pi-i. In summary,the population composition of M. oryzae in Jiangsu province is relatively complex,and it comes instable. The distribution of avr-genes is quite different and closely related to the geographical area.
引文
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[2]周萌,邓国富,梁海福,等.抗稻瘟病优质籼型水稻不育系青A选育及应用[J].南方农业学报,2017,48(1):26-30.
[3] QI Z Q,LIU M X,ZHANG Z G,et al. The syntaxin protein(Mo-Syn8)mediates intracellular trafficking to regulate conidiogenesisand pathogenicity of rice blast fungus[J]. New Phytologist,2016,209(4):1655-1667.
[4]宋兆强,刘艳,王宝祥,等.稻瘟病抗性基因Pi-ta、Pi-b、Pi54和Pi-km的育种利用价值评价[J].江苏农业学报,2017,33(5):968-974.
[5]阎勇,马增凤,秦钢,等.华南常用籼稻亲本稻瘟病抗性评价及抗性基因鉴定[J].南方农业学报,2017,48(4):587-593.
[6]马军韬,张国民,张丽艳,等.黑龙江省水稻品种抗性与稻瘟病病菌致病性年际变化趋势分析[J].江苏农业科学,2017,45(20):109-113.
[7]马军韬,张国民,张丽艳,等.黑龙江省不同来源稻瘟病菌致病性分析[J].植物保护,2017,43(1):161-164.
[8]张善磊,孙旭超,陈涛,等. Pi-ta、Pi-5、Pi-km和Pi-b基因在粳稻品种(系)中的分布及对穗颈瘟的抗性[J].江苏农业学报,2018,34(5):961-971.
[9]姜兆远,刘晓梅,李莉,等.吉林省稻瘟病菌无毒基因Avr Pi9鉴定及分析[J].江苏农业科学,2017,45(10):27-29.
[10]杨军,薛芳,王海凤,等.水稻稻瘟病菌单孢分离技术及常见问题分析[J].山东农业科学,2017,49(2):132-135.
[11] FARMAN M,LEONG S. Chromosome walking to the AVR1-CO39avirulence gene of Magnaporthe grisea:discrepancy between thephysical and genetic maps[J]. Genetics,1998,150:1049-1058.
[12] WU J,KOU YJ,BAO JD,et al. Comparative genomics identifiesthe Magnaporthe oryzae avirulence effector Avr Pi9 that triggersPi9-mediated blast resistance in rice[J]. New Phygologist,2015,206(4):1463-1475.
[13] LI W,WANG B,ZHANG X,et al. The Magnaporthe oryzae avir-ulence gene Avr Piz-t encodes a predicted secreted protein that trig-gers the immunity in rice mediated by the blast resistance gene Piz-t[J]. Molecular Plant Microbe Interaction,2009,22:411-420.
[14]张哲,姜华,孙国昌,等.稻瘟病菌无毒基因研究进展[J].遗传,2011,33(6):591-600.
[15] DONG Y H,LI Y,ZHAO M M,et al. Global genome and tran-scriptome analysis of Magnaporthe oryzae epidemic isolate 98-06uncover novel effectors and pathogenicity-related genes,revealinggene gain and lose dynamics in genome evolution[J]. PLoSPathogens,2015,11(4):e1004801.
[16] QI Z Q,WANG Q,DOU X Y,et al. Mo Swi6,an APSES familytranscription factor,interacts with Mo Mps1 and is required for hy-phal and conidial morphogenesis,appressorial function and patho-genicity of Magnaporthe oryzae[J]. Molecular Plant Pathology,2012,13(7):677-689.
[17] ZHANG S,WANG L,PAN Q,et al. Functional evaluation ofMagnaporthe oryzae avirulence gene Avr Pib responding to the riceblast resistance gene Pib[J]. Scientific Reports,2015,5:11642.
[18] FLOR H. Current status of the gene-for-gene concept[J]. AnnualReview of Phytopathology,1971,9:275-296.
[19] DENG Y,ZHAI K,XIE Z,et al. Epigenetic regulation of antago-nistic receptors confers rice blast resistance with yield balance[J].Science,2017,355(6328):962-965.
[20] DE WIT P J G. How plants recognize pathogens and defend them-selves[J]. Cell&Molecular Life Sciences Cmls,2007,64(21):2726-2732.
[21]李祥晓,王倩,黎志康,等.黑龙江省稻瘟病菌无毒基因分析及抗病种质资源筛选[J].作物学报,2012,38(12):2192-2197.
[22]兰波,李湘民,何烈干.江西省稻瘟病菌的无毒基因分析[J].江西农业大学学报,2010,32(2):271-275.
[23]王世维,郑文静,刘志恒,等.辽宁省稻瘟病菌无毒基因型鉴定及分析[J].中国农业科学,2014,47(3):462-472.
[24]杨秀娟,阮宏椿,王茂明,等.福建省稻瘟病菌致病性及其无毒基因分析[J].植物保护学报,2007,34(4):337-342.
[25]凌忠专,雷财林,王久林.稻瘟病菌生理小种研究的回顾与展望[J].中国农业科学,2004,37(12):1849-1859.
[26]周江鸿,王久林,凌忠专,等.我国稻瘟病菌毒力基因的组成及其地理分布[J].作物学报,2003,29(5):646-651.
[27]张科,刘丽,刘东渤,等.黑龙江省稻瘟病菌无毒基因的组成及其频率初探[J].种子世界,2016(12):19-21.
[28]朱名海,赵美,舒灿伟,等.南繁区稻瘟病菌无毒基因的检测[J].华中农业大学学报,2017,36(4):21-25.
[29]张琦峰,靳学慧,蔡鑫鑫,等.黑龙江省稻瘟病菌无毒基因的检测[J].黑龙江农业科学,2014(12):70-73.
[30] VALENT B,CHUMLEY F. Molecular genetic analysis of the riceblast fungus Magnaporthe grisea[J]. Annual Review of Phytopa-thology,1991,29:443-467.
[2]周萌,邓国富,梁海福,等.抗稻瘟病优质籼型水稻不育系青A选育及应用[J].南方农业学报,2017,48(1):26-30.
[3] QI Z Q,LIU M X,ZHANG Z G,et al. The syntaxin protein(Mo-Syn8)mediates intracellular trafficking to regulate conidiogenesisand pathogenicity of rice blast fungus[J]. New Phytologist,2016,209(4):1655-1667.
[4]宋兆强,刘艳,王宝祥,等.稻瘟病抗性基因Pi-ta、Pi-b、Pi54和Pi-km的育种利用价值评价[J].江苏农业学报,2017,33(5):968-974.
[5]阎勇,马增凤,秦钢,等.华南常用籼稻亲本稻瘟病抗性评价及抗性基因鉴定[J].南方农业学报,2017,48(4):587-593.
[6]马军韬,张国民,张丽艳,等.黑龙江省水稻品种抗性与稻瘟病病菌致病性年际变化趋势分析[J].江苏农业科学,2017,45(20):109-113.
[7]马军韬,张国民,张丽艳,等.黑龙江省不同来源稻瘟病菌致病性分析[J].植物保护,2017,43(1):161-164.
[8]张善磊,孙旭超,陈涛,等. Pi-ta、Pi-5、Pi-km和Pi-b基因在粳稻品种(系)中的分布及对穗颈瘟的抗性[J].江苏农业学报,2018,34(5):961-971.
[9]姜兆远,刘晓梅,李莉,等.吉林省稻瘟病菌无毒基因Avr Pi9鉴定及分析[J].江苏农业科学,2017,45(10):27-29.
[10]杨军,薛芳,王海凤,等.水稻稻瘟病菌单孢分离技术及常见问题分析[J].山东农业科学,2017,49(2):132-135.
[11] FARMAN M,LEONG S. Chromosome walking to the AVR1-CO39avirulence gene of Magnaporthe grisea:discrepancy between thephysical and genetic maps[J]. Genetics,1998,150:1049-1058.
[12] WU J,KOU YJ,BAO JD,et al. Comparative genomics identifiesthe Magnaporthe oryzae avirulence effector Avr Pi9 that triggersPi9-mediated blast resistance in rice[J]. New Phygologist,2015,206(4):1463-1475.
[13] LI W,WANG B,ZHANG X,et al. The Magnaporthe oryzae avir-ulence gene Avr Piz-t encodes a predicted secreted protein that trig-gers the immunity in rice mediated by the blast resistance gene Piz-t[J]. Molecular Plant Microbe Interaction,2009,22:411-420.
[14]张哲,姜华,孙国昌,等.稻瘟病菌无毒基因研究进展[J].遗传,2011,33(6):591-600.
[15] DONG Y H,LI Y,ZHAO M M,et al. Global genome and tran-scriptome analysis of Magnaporthe oryzae epidemic isolate 98-06uncover novel effectors and pathogenicity-related genes,revealinggene gain and lose dynamics in genome evolution[J]. PLoSPathogens,2015,11(4):e1004801.
[16] QI Z Q,WANG Q,DOU X Y,et al. Mo Swi6,an APSES familytranscription factor,interacts with Mo Mps1 and is required for hy-phal and conidial morphogenesis,appressorial function and patho-genicity of Magnaporthe oryzae[J]. Molecular Plant Pathology,2012,13(7):677-689.
[17] ZHANG S,WANG L,PAN Q,et al. Functional evaluation ofMagnaporthe oryzae avirulence gene Avr Pib responding to the riceblast resistance gene Pib[J]. Scientific Reports,2015,5:11642.
[18] FLOR H. Current status of the gene-for-gene concept[J]. AnnualReview of Phytopathology,1971,9:275-296.
[19] DENG Y,ZHAI K,XIE Z,et al. Epigenetic regulation of antago-nistic receptors confers rice blast resistance with yield balance[J].Science,2017,355(6328):962-965.
[20] DE WIT P J G. How plants recognize pathogens and defend them-selves[J]. Cell&Molecular Life Sciences Cmls,2007,64(21):2726-2732.
[21]李祥晓,王倩,黎志康,等.黑龙江省稻瘟病菌无毒基因分析及抗病种质资源筛选[J].作物学报,2012,38(12):2192-2197.
[22]兰波,李湘民,何烈干.江西省稻瘟病菌的无毒基因分析[J].江西农业大学学报,2010,32(2):271-275.
[23]王世维,郑文静,刘志恒,等.辽宁省稻瘟病菌无毒基因型鉴定及分析[J].中国农业科学,2014,47(3):462-472.
[24]杨秀娟,阮宏椿,王茂明,等.福建省稻瘟病菌致病性及其无毒基因分析[J].植物保护学报,2007,34(4):337-342.
[25]凌忠专,雷财林,王久林.稻瘟病菌生理小种研究的回顾与展望[J].中国农业科学,2004,37(12):1849-1859.
[26]周江鸿,王久林,凌忠专,等.我国稻瘟病菌毒力基因的组成及其地理分布[J].作物学报,2003,29(5):646-651.
[27]张科,刘丽,刘东渤,等.黑龙江省稻瘟病菌无毒基因的组成及其频率初探[J].种子世界,2016(12):19-21.
[28]朱名海,赵美,舒灿伟,等.南繁区稻瘟病菌无毒基因的检测[J].华中农业大学学报,2017,36(4):21-25.
[29]张琦峰,靳学慧,蔡鑫鑫,等.黑龙江省稻瘟病菌无毒基因的检测[J].黑龙江农业科学,2014(12):70-73.
[30] VALENT B,CHUMLEY F. Molecular genetic analysis of the riceblast fungus Magnaporthe grisea[J]. Annual Review of Phytopa-thology,1991,29:443-467.