马铃薯晚疫病菌有性生殖发生对无毒基因Avr-blb1多样性及菌株毒性的影响
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摘要
【目的】马铃薯晚疫病菌(Phytophthora infestans)是马铃薯重要病害病原,前期研究表明,有性生殖的发生是群体结构变异的重要手段。【方法】在云南省马铃薯主产区采集晚疫病菌,分离纯化19株,其中A2交配型15株,是优势群体,A1A2交配型2株,A1交配型1株,自育型(SF) 1株;对来自不同寄主品种、不同交配型的4株菌株进行有性生殖诱导,以及对有性后代进行毒性测定。【结果】确定了3对卵孢子产生较多的亲本组合;采用改良的卵孢子萌发诱导方法得到的卵孢子有性后代菌株30株,卵孢子萌发率达6%。在有性后代群体中Avr-blb1无毒基因序列发生了多态性分离,其中亲本组合1、组合2和组合3的有性生殖后代序列多态性系数分别为0. 013,0. 010和0. 002;通过对马铃薯栽培品种合作88的毒性验证,发现组合1和组合3后代均有毒性分离现象。【结论】上述结果表明了有性生殖发生导致的无毒基因序列多样性对病原菌的毒性变异具有重要作用。
【Objective】Phytophthora infestans,cause of the potato late blight disease,was one of the most infamous diseases in agriculture.Previous studies showed that the occurrence of sexual reproduction was an important reason of population structure variation.【Method】Nineteen P. infestans isolates were collected and purified in the major potato producing area of Yunnan province. Among them,15 isolates were identified as A2 mating type,was the dominant groups,meanwhile,2 isolates of A1 A2,each 1 isolate of self-fertile( SF) and A1 mating type were also found. Four isolates representing different mating type collected from various hosts were used for inducing sexual reproduction and pathogenicity test.【Result】Oospore germination rate up to 6 % by optimized inducing methods and 30 sexual F1 offspring were obtained from 3 parental paired treatments,sequence analysis of the Avr-blb1 ORF were detected DNA polymorphism diversity,the genetic diversity index equal to 0. 01301,0. 01023 and 0. 00219,respectively. Phenotypic test on potato variety cooperation 88 showed pathogenicity segregation between parental paired treatment 1 and 3.【Conclusion】These results demonstrated that the occurrence of sexual reproduction in P. infestans had an important role in Avr gene and pathogenicity variation.
【Objective】Phytophthora infestans,cause of the potato late blight disease,was one of the most infamous diseases in agriculture.Previous studies showed that the occurrence of sexual reproduction was an important reason of population structure variation.【Method】Nineteen P. infestans isolates were collected and purified in the major potato producing area of Yunnan province. Among them,15 isolates were identified as A2 mating type,was the dominant groups,meanwhile,2 isolates of A1 A2,each 1 isolate of self-fertile( SF) and A1 mating type were also found. Four isolates representing different mating type collected from various hosts were used for inducing sexual reproduction and pathogenicity test.【Result】Oospore germination rate up to 6 % by optimized inducing methods and 30 sexual F1 offspring were obtained from 3 parental paired treatments,sequence analysis of the Avr-blb1 ORF were detected DNA polymorphism diversity,the genetic diversity index equal to 0. 01301,0. 01023 and 0. 00219,respectively. Phenotypic test on potato variety cooperation 88 showed pathogenicity segregation between parental paired treatment 1 and 3.【Conclusion】These results demonstrated that the occurrence of sexual reproduction in P. infestans had an important role in Avr gene and pathogenicity variation.
引文
[1]Fry W. Phytophthora infestans:the plant(and R gene)destroyer[J]. Molecular Plant Pathology,2008,9(3):385-402.
[2]Halterman D A,Chen Y,Sopee J,et al. Competition between Phy-tophthora infestans effectors leads to increased aggressiveness onplants containing broad-spectrum late blight resistance[J]. PLo SOne,2010,5(5):e10536.
[3]Song J,Bradeen J M,Naess S K,et al. Gene RB cloned from Sola-num bulbocastanum confers broad spectrum resistance to potato lateblight[J]. Proceedings of the National Academy of Sciences,2003,100(16):9128-9133.
[4]Burdon J J,Thrall P H. The fitness costs to plants of resistance topathogens[J]. Genome Biology,2003,4(9):227.
[5]van de Vondervoort P J I,Bouwmeester K,Govers F. Allele varia-tion in the Phytophthora infestans effector gene ipiO[C]//Book ofAbstracts Annual meeting Oomycete Molecular Genetics Network,Wageningen,The Netherlands,2006:40(30).
[6]赵志坚,张志铭.云南省发现马铃薯晚疫病菌(Phytophthora in-festans)A2交配型[J].西南农业学报,1999,12(3):1-3.
[7]杨素祥.云南马铃薯晚疫病菌群体的遗传多样性研究[D].昆明:云南师范大学,2006.
[8]孟然然,郝大海,王兆富,等.马铃薯晚疫病菌SSR扩增产物的检测方法比较[J].安徽农学通报,2012,18(13):23-24.
[9]朱杰华.中国马铃薯晚疫病菌群体遗传结构研究[D].保定:河北农业大学,2004.
[10]Yuen J E,Andersson B. What is the evidence for sexual reproduc-tion of Phytophthora infestans in Europe?[J]. Plant Pathology,2013,62(3):485-491.
[11]Judelson H S. Genetic and physical variability at the mating type lo-cus of the oomycete,Phytophthora infestans[J]. Genetics,1996,144(3):1005-1013.
[12]Fabritius A L,Cvitanich C,Judelson H S. Stage-specific gene ex-pression during sexual development in Phytophthora infestans[J].Molecular Microbiology,2002,45(4):1057-1066.
[13]马利松.马铃薯晚疫病菌卵孢子生物学特性的研究[D].保定:河北农业大学,2003.
[14]Chang T T,Ko W H. Factors affecting germination of oospores ofPhytophthora infestans[J]. Journal of Phytopathology,1991,133(1):29-35.
[15]Yuen J E,Andersson B. What is the evidence for sexual reproduc-tion of Phytophthora infestans in Europe?[J]. Plant Pathology,2013,62(3):485-491.
[16]杨芮,方治国,詹家绥,等.马铃薯晚疫病病菌在贵州和云南的交配型分布与卵孢子生物学特性分析[J].江苏农业科学,2014,42(7):122-124.
[17]Li Y,van der Lee T A J,Evenhuis A,et al. Population dynamicsof Phytophthora infestans in the Netherlands reveals expansion andspread of dominant clonal lineages and virulence in sexual offspring[J]. G3:Genes,Genomes,Genetics,2012,2(12):1529-1540.
[18]Halterman D A,Chen Y,Sopee J,et al. Competition betweenPhytophthora infestans effectors leads to increased aggressiveness onplants containing broad-spectrum late blight resistance[J]. PLo SOne,2010,5(5):e10536.
[19]Champouret N,Rietman H,Bouwmeester K,et al.(A)virulenceon Rpi-blb1 expressing plants is determined by the ipiO effector fami-ly from Phytophthora infestans[C]//Book of Abstracts Annual Meet-ing Oomycete Molecular Genetics Network,Birnam,Scotland,Unit-ed Kingdom,2008:23.
[20]Champouret N,Bouwmeester K,Rietman H,et al. Phytophthorainfestans isolates lacking class I ipiO variants are virulent on Rpi-blb1potato[J]. Molecular Plant-microbe Interactions,2009,22(12):1535-1545.
[21]Nyongesa M,Phelan S,Wright D,et al. Evaluating the potential ofPhytophthora infestans to genetically adapt to the Rpi-blb1(RB)source of blight resistance[C]//Proceedings of the Fourteenth Eu-ro Blight Workshop,Limassol,Cyprus,2014:163-168.
[22]Pieterse C M J,van West P,Verbakel H M,et al. Structure andgenomic organization of the ipi B and ipiO gene clusters of Phytoph-thora infestans[J]. Gene,1994,138(1):67-77.
[23]Pieterse C M J,Riach M B R,Bleker T,et al. Isolation of putativepathogenicity genes of the potato late blight fungus Phytophthora in-festans by differential hybridization of a genomic library[J]. Physio-logical and Molecular Plant Pathology,1993,43(1):69-79.
[2]Halterman D A,Chen Y,Sopee J,et al. Competition between Phy-tophthora infestans effectors leads to increased aggressiveness onplants containing broad-spectrum late blight resistance[J]. PLo SOne,2010,5(5):e10536.
[3]Song J,Bradeen J M,Naess S K,et al. Gene RB cloned from Sola-num bulbocastanum confers broad spectrum resistance to potato lateblight[J]. Proceedings of the National Academy of Sciences,2003,100(16):9128-9133.
[4]Burdon J J,Thrall P H. The fitness costs to plants of resistance topathogens[J]. Genome Biology,2003,4(9):227.
[5]van de Vondervoort P J I,Bouwmeester K,Govers F. Allele varia-tion in the Phytophthora infestans effector gene ipiO[C]//Book ofAbstracts Annual meeting Oomycete Molecular Genetics Network,Wageningen,The Netherlands,2006:40(30).
[6]赵志坚,张志铭.云南省发现马铃薯晚疫病菌(Phytophthora in-festans)A2交配型[J].西南农业学报,1999,12(3):1-3.
[7]杨素祥.云南马铃薯晚疫病菌群体的遗传多样性研究[D].昆明:云南师范大学,2006.
[8]孟然然,郝大海,王兆富,等.马铃薯晚疫病菌SSR扩增产物的检测方法比较[J].安徽农学通报,2012,18(13):23-24.
[9]朱杰华.中国马铃薯晚疫病菌群体遗传结构研究[D].保定:河北农业大学,2004.
[10]Yuen J E,Andersson B. What is the evidence for sexual reproduc-tion of Phytophthora infestans in Europe?[J]. Plant Pathology,2013,62(3):485-491.
[11]Judelson H S. Genetic and physical variability at the mating type lo-cus of the oomycete,Phytophthora infestans[J]. Genetics,1996,144(3):1005-1013.
[12]Fabritius A L,Cvitanich C,Judelson H S. Stage-specific gene ex-pression during sexual development in Phytophthora infestans[J].Molecular Microbiology,2002,45(4):1057-1066.
[13]马利松.马铃薯晚疫病菌卵孢子生物学特性的研究[D].保定:河北农业大学,2003.
[14]Chang T T,Ko W H. Factors affecting germination of oospores ofPhytophthora infestans[J]. Journal of Phytopathology,1991,133(1):29-35.
[15]Yuen J E,Andersson B. What is the evidence for sexual reproduc-tion of Phytophthora infestans in Europe?[J]. Plant Pathology,2013,62(3):485-491.
[16]杨芮,方治国,詹家绥,等.马铃薯晚疫病病菌在贵州和云南的交配型分布与卵孢子生物学特性分析[J].江苏农业科学,2014,42(7):122-124.
[17]Li Y,van der Lee T A J,Evenhuis A,et al. Population dynamicsof Phytophthora infestans in the Netherlands reveals expansion andspread of dominant clonal lineages and virulence in sexual offspring[J]. G3:Genes,Genomes,Genetics,2012,2(12):1529-1540.
[18]Halterman D A,Chen Y,Sopee J,et al. Competition betweenPhytophthora infestans effectors leads to increased aggressiveness onplants containing broad-spectrum late blight resistance[J]. PLo SOne,2010,5(5):e10536.
[19]Champouret N,Rietman H,Bouwmeester K,et al.(A)virulenceon Rpi-blb1 expressing plants is determined by the ipiO effector fami-ly from Phytophthora infestans[C]//Book of Abstracts Annual Meet-ing Oomycete Molecular Genetics Network,Birnam,Scotland,Unit-ed Kingdom,2008:23.
[20]Champouret N,Bouwmeester K,Rietman H,et al. Phytophthorainfestans isolates lacking class I ipiO variants are virulent on Rpi-blb1potato[J]. Molecular Plant-microbe Interactions,2009,22(12):1535-1545.
[21]Nyongesa M,Phelan S,Wright D,et al. Evaluating the potential ofPhytophthora infestans to genetically adapt to the Rpi-blb1(RB)source of blight resistance[C]//Proceedings of the Fourteenth Eu-ro Blight Workshop,Limassol,Cyprus,2014:163-168.
[22]Pieterse C M J,van West P,Verbakel H M,et al. Structure andgenomic organization of the ipi B and ipiO gene clusters of Phytoph-thora infestans[J]. Gene,1994,138(1):67-77.
[23]Pieterse C M J,Riach M B R,Bleker T,et al. Isolation of putativepathogenicity genes of the potato late blight fungus Phytophthora in-festans by differential hybridization of a genomic library[J]. Physio-logical and Molecular Plant Pathology,1993,43(1):69-79.