马铃薯高世代选系疮痂病抗性评价及SSR辅助筛选
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摘要
利用国外种质资源创造的20份高世代马铃薯无性系为试材,以尤金为感病对照,克新18号为抗病对照,采用自然病圃和人工接种的鉴定方式进行疮痂病抗性评价,利用SSR分子标记分析试材之间的亲缘关系,并对比SSR遗传相似性系数聚类结果和试材抗病性分类结果,探究利用SSR分子标记辅助筛选疮痂病抗病资源的可行性。从试验地土壤和感病块茎共分离纯化具有链霉菌特征的菌株278份,其中240份来自于块茎,38份来自于土壤。具有致病力的菌株127株,经鉴定全部属于Streptomyces scabies菌种。试材间的抗病性存在明显差异,可分为高抗、中抗、中感和高感4个类型。人工接种与自然病圃抗性鉴定结果存在极显著相关(R~2=0.946 7)。抗病类型与感病类型通过SSR遗传相似性系数聚类大致可以区分。
Taking 20 potato advanced generation clones created by foreign germplasm resources as test material,‘Youjin'as susceptible contrast and‘Kexin No.18' as resistant contrast,this paper evaluated the scab resistance of test material by natural infested field and artificial inoculation methods;analyzed the genetic relationship between test materials using SSR molecular markers;and compared the clustering results of SSR genetic similarity coefficient and classification results of disease resistance of the tested materials,so as to explore the feasibility of using SSR molecular marker for screening resources with scab resistance.Two hundred and seventy-eight bacterial strains with Streptomyces characteristics were separated and purified from soil and tuber,of which 240 from tuber and 38 from soil.One hundred and twenty-seven bacterial strains with pathogenicity were all identified as Streptomyces scabies.There were significant differences in disease resistance among the tested materials,which could be divided into 4 types:high resistant,moderate resistant,moderate susceptible and high susceptible.Correlation between artificial inoculation and resistance identification in natural disease nurseries was highly significant(R~2=0.946 7).The resistant type and susceptible type could be roughly distinguished by SSR genetic similarity coefficient clustering,indicating that scab resistance was related to the genetic relationship.
Taking 20 potato advanced generation clones created by foreign germplasm resources as test material,‘Youjin'as susceptible contrast and‘Kexin No.18' as resistant contrast,this paper evaluated the scab resistance of test material by natural infested field and artificial inoculation methods;analyzed the genetic relationship between test materials using SSR molecular markers;and compared the clustering results of SSR genetic similarity coefficient and classification results of disease resistance of the tested materials,so as to explore the feasibility of using SSR molecular marker for screening resources with scab resistance.Two hundred and seventy-eight bacterial strains with Streptomyces characteristics were separated and purified from soil and tuber,of which 240 from tuber and 38 from soil.One hundred and twenty-seven bacterial strains with pathogenicity were all identified as Streptomyces scabies.There were significant differences in disease resistance among the tested materials,which could be divided into 4 types:high resistant,moderate resistant,moderate susceptible and high susceptible.Correlation between artificial inoculation and resistance identification in natural disease nurseries was highly significant(R~2=0.946 7).The resistant type and susceptible type could be roughly distinguished by SSR genetic similarity coefficient clustering,indicating that scab resistance was related to the genetic relationship.
引文
何虎翼，谭冠宁，何新民，唐洲萍，李丽淑，杨鑫．2017．马铃薯品种（系）资源的疮痂病抗性鉴定．植物遗传资源学报，18(4):786-793．
靳海波，王文丽，邱慧珍，李娟，赵旭．2015．生物有机肥GSJ-1对马铃薯土壤疮痂病病原菌分布影响及生防效果研究．干旱地区农业研究，33(2):165-169．
刘思泱．2010.6个彩色马铃薯品种的核型及ISSR分析[硕士论文]．呼和浩特：内蒙古农业大学．
聂峰杰，陈虞超，巩檑，张丽，甘晓燕，石磊，宋玉霞．2018．马铃薯疮痂病致病链霉菌分类及其致病机理研究进展．分子植物育种，（4):1313-1319．
单友蛟，刘杰，卞春松，段绍光，徐建飞，庞万福，金黎平．2010．马铃薯SSR遗传连锁图谱构建及3个重要农艺性状QTLs定位．中国蔬菜，（18):10-14．
石景．2011．基于SSR标记的彩色马铃薯亲缘关系分析及指纹图谱构建[硕士论文]．武汉：华中农业大学．
孙静，金光辉，刘喜才．2015．不同药剂及施用方式对马铃薯疮痂病的防效．中国马铃薯，（2):107-111．
王跃栋，刘自学，苏爱莲．2011．草地早熟禾品种在北京地区对褐斑病的抗性评价．草业科学，28(10):1796-1800．
魏延安．2005．世界马铃薯产业发展现状及特点说明．现代种业，（3):29-32．
吴立萍．2017．马铃薯种质资源的疮痂病抗性鉴定及其抗源的遗传多样性分析[硕士论文]．大庆：黑龙江八一农垦大学．
邢莹莹．2015．黑龙江省部分地区马铃薯疮痂病病原鉴定及主栽品种的抗性评价[硕士论文]．大庆：黑龙江八一农垦大学．
徐建飞，金黎平．2017．马铃薯遗传育种研究：现状与展望．中国农业科学，50(6):990-1015．
张建平，哈斯，林团荣，尹玉和，郑友，闫任沛，程玉臣，胡冰．2013．不同杀菌剂对马铃薯疮痂病的防效试验．中国马铃薯，27(2):83-86．
张萌，赵伟全，于秀梅，杨文香，张汀，刘大群．2009．中国马铃薯疮痂病病原菌16S rDNA的遗传多样性分析．中国农业科学，42(2):499-504．
Jan F K,Sini S,Lars P,Valkonen J P.1999.Phylogenetic analysis of 16S rRNA genes and PCR analysis of the nec1 gene from Streptomyces spp.causing common scab,pitted scab,and netted scab in Finland.Phytopathology,89(6):462-469．
Lehtonen M J,Rantala H,Kreuze J F,Bang H,Kuisma L,Koski P,Virtanen E,Vihlman K,Valkonen J P T.2004.Occurrence and survival of potato scab pathogens(Streptomyces species)on tuber lesions:quick diagnosis based on a PCR-based assay.Plant Pathology,53(3):280-287．
Mishra K K,Srivastava J S.2004.Soil amendments to control common scab of potato.Potato Research,47(1):101-109．
Pasco C,Jouan B,Andrivon D.2005.Resistance of potato genotypes to common and netted scab-causing species of Streptomyces.Plant Pathology,54(3):383-392．
Singhai P K,Sarma B K,Srivastava J S.2011.Phenolic acid content in potato peel determines natural infection of common scab caused by Streptomyces spp.World Journal of Microbiology&Biotechnology,27(7):1559-1567．
Waterer D.2002.Impact of high soil pH on potato fields and grade losses to common scab.Canadian Journal of Plant Science,82(3):583-586．
Wanner L A.2006.A survey of genetic variation in Streptomyces isolates causing potato common scab in the United States.Phytopathology,96(12):1363-1371．
Wanner L A,Haynes K G.2009.Aggressiveness of Streptomyces on four potato cultivars and implications for common scab resistance breeding.American Journal Potato Research,86(5):335-346.
靳海波，王文丽，邱慧珍，李娟，赵旭．2015．生物有机肥GSJ-1对马铃薯土壤疮痂病病原菌分布影响及生防效果研究．干旱地区农业研究，33(2):165-169．
刘思泱．2010.6个彩色马铃薯品种的核型及ISSR分析[硕士论文]．呼和浩特：内蒙古农业大学．
聂峰杰，陈虞超，巩檑，张丽，甘晓燕，石磊，宋玉霞．2018．马铃薯疮痂病致病链霉菌分类及其致病机理研究进展．分子植物育种，（4):1313-1319．
单友蛟，刘杰，卞春松，段绍光，徐建飞，庞万福，金黎平．2010．马铃薯SSR遗传连锁图谱构建及3个重要农艺性状QTLs定位．中国蔬菜，（18):10-14．
石景．2011．基于SSR标记的彩色马铃薯亲缘关系分析及指纹图谱构建[硕士论文]．武汉：华中农业大学．
孙静，金光辉，刘喜才．2015．不同药剂及施用方式对马铃薯疮痂病的防效．中国马铃薯，（2):107-111．
王跃栋，刘自学，苏爱莲．2011．草地早熟禾品种在北京地区对褐斑病的抗性评价．草业科学，28(10):1796-1800．
魏延安．2005．世界马铃薯产业发展现状及特点说明．现代种业，（3):29-32．
吴立萍．2017．马铃薯种质资源的疮痂病抗性鉴定及其抗源的遗传多样性分析[硕士论文]．大庆：黑龙江八一农垦大学．
邢莹莹．2015．黑龙江省部分地区马铃薯疮痂病病原鉴定及主栽品种的抗性评价[硕士论文]．大庆：黑龙江八一农垦大学．
徐建飞，金黎平．2017．马铃薯遗传育种研究：现状与展望．中国农业科学，50(6):990-1015．
张建平，哈斯，林团荣，尹玉和，郑友，闫任沛，程玉臣，胡冰．2013．不同杀菌剂对马铃薯疮痂病的防效试验．中国马铃薯，27(2):83-86．
张萌，赵伟全，于秀梅，杨文香，张汀，刘大群．2009．中国马铃薯疮痂病病原菌16S rDNA的遗传多样性分析．中国农业科学，42(2):499-504．
Jan F K,Sini S,Lars P,Valkonen J P.1999.Phylogenetic analysis of 16S rRNA genes and PCR analysis of the nec1 gene from Streptomyces spp.causing common scab,pitted scab,and netted scab in Finland.Phytopathology,89(6):462-469．
Lehtonen M J,Rantala H,Kreuze J F,Bang H,Kuisma L,Koski P,Virtanen E,Vihlman K,Valkonen J P T.2004.Occurrence and survival of potato scab pathogens(Streptomyces species)on tuber lesions:quick diagnosis based on a PCR-based assay.Plant Pathology,53(3):280-287．
Mishra K K,Srivastava J S.2004.Soil amendments to control common scab of potato.Potato Research,47(1):101-109．
Pasco C,Jouan B,Andrivon D.2005.Resistance of potato genotypes to common and netted scab-causing species of Streptomyces.Plant Pathology,54(3):383-392．
Singhai P K,Sarma B K,Srivastava J S.2011.Phenolic acid content in potato peel determines natural infection of common scab caused by Streptomyces spp.World Journal of Microbiology&Biotechnology,27(7):1559-1567．
Waterer D.2002.Impact of high soil pH on potato fields and grade losses to common scab.Canadian Journal of Plant Science,82(3):583-586．
Wanner L A.2006.A survey of genetic variation in Streptomyces isolates causing potato common scab in the United States.Phytopathology,96(12):1363-1371．
Wanner L A,Haynes K G.2009.Aggressiveness of Streptomyces on four potato cultivars and implications for common scab resistance breeding.American Journal Potato Research,86(5):335-346.