抗根肿病甘蓝型油菜新种质的创制及抗性评价
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摘要
【目的】通过根肿病高抗白菜品种"康根51"与甘蓝型油菜种质进行杂交,创制获得对云南特定根肿病生理小种具有抗性的种质,为根肿病抗性育种奠定基础。【方法】采用染色体数目和PCR扩增对亲本和杂交后代进行分析,并采用田间抗性鉴定和室内人工接种鉴定相结合的方法进行抗性评价。【结果】染色体数目分析发现,"康根51"的染色体数目为20条,油菜10159染色体数目为38条,杂交F_1代材料染色体数目为29条,与预期结果相符。随着世代的增加,染色体数目逐渐趋于正常且自交结实正常。PCR扩增结果显示杂交后代具有双亲的互补带型,表明创制的种质为甘、白杂交后代种质。对不同世代的663份自交单株进行田间根肿病抗性初筛,29份材料发病率较低;以楚雄东瓜镇油菜发病根瘤上分离的休眠孢子为病原对田间初筛的种质进行室内抗性鉴定。R150和R322表现为高抗,发病率分别为6.7%和13.3%,病情指数分别为5.83和7.50;R072、R087等19份材料表现为抗病,发病率在13.3%～36.7%,病情指数在10.83～25.80,其余8份表现为感病或高感。相关性分析表明,油菜根肿病的病情指数与发病率呈显著正相关。【结论】获得对云南特定根肿病生理小种表现抗性的甘蓝型油菜新种质2份,可用于油菜根肿病抗病育种。
【Objective】To obtain clubroot-resistant Brassica napus germplasm in Yunnan and lay a foundation for resistance breeding, clubroot-resistant Chinese cabbage variety ‘CR51' was crossed with clubroot-susceptible Brassica napus.【Method】Chromosome numbers and PCR amplification were conducted to analyze the parents and filial generations, and the method of field resistance identification and artificial inoculation was used to evaluate the resistance of parents and progenies.【Result】Chromosome numbers analysis showed that the number of ‘CR51' was 20, rapeseed 10159 was 38 and the number of F_1 was 29, consistent with the expected results. With the increasing of generation, the number of chromosome and self-fruitfulness tended to be normal. PCR amplification results showed that the F_2 hybrid possessed the complementary bands of parents, which indicated that they were hybrids of Chinese cabbage and rapeseed. 29 offspring at different generations with low disease incidence were selected by field clubroot resistance identification. Clubroot pathogen obtained from Donggua Town of Chuxiong was used to artificial inoculation, which showed that R150 and R322 were highly resistant to clubroot, the disease incidence was 6.7 % and 13.3 %, the disease index was 5.83 and 7.50, respectively. 19 germplasm such as R072, R087 were resistant to clubroot, and the disease incidence was between 13.3 % and 36.7 % and the disease index was between 10.83 and 25.80.The rest 8 germplasm were susceptible or highly susceptible to clubroot. Correlation analysis showed that there was a significant positive correlation between the disease index and incidence of clubroot in brassica napus.【Conclusion】 Two new germplasm of Brassica napus highly resistant to certain physiological race in Yunnan were obtained via interbreeding which could be used in rapeseed clubroot-resistance breeding.
【Objective】To obtain clubroot-resistant Brassica napus germplasm in Yunnan and lay a foundation for resistance breeding, clubroot-resistant Chinese cabbage variety ‘CR51' was crossed with clubroot-susceptible Brassica napus.【Method】Chromosome numbers and PCR amplification were conducted to analyze the parents and filial generations, and the method of field resistance identification and artificial inoculation was used to evaluate the resistance of parents and progenies.【Result】Chromosome numbers analysis showed that the number of ‘CR51' was 20, rapeseed 10159 was 38 and the number of F_1 was 29, consistent with the expected results. With the increasing of generation, the number of chromosome and self-fruitfulness tended to be normal. PCR amplification results showed that the F_2 hybrid possessed the complementary bands of parents, which indicated that they were hybrids of Chinese cabbage and rapeseed. 29 offspring at different generations with low disease incidence were selected by field clubroot resistance identification. Clubroot pathogen obtained from Donggua Town of Chuxiong was used to artificial inoculation, which showed that R150 and R322 were highly resistant to clubroot, the disease incidence was 6.7 % and 13.3 %, the disease index was 5.83 and 7.50, respectively. 19 germplasm such as R072, R087 were resistant to clubroot, and the disease incidence was between 13.3 % and 36.7 % and the disease index was between 10.83 and 25.80.The rest 8 germplasm were susceptible or highly susceptible to clubroot. Correlation analysis showed that there was a significant positive correlation between the disease index and incidence of clubroot in brassica napus.【Conclusion】 Two new germplasm of Brassica napus highly resistant to certain physiological race in Yunnan were obtained via interbreeding which could be used in rapeseed clubroot-resistance breeding.
引文
[1]Piao Z Y,Ramchiary N,Lim Y P.Genetics of clubroot resistance in Brassica species[J].Journal of Plant Growth Regulation,2009,28:252-264.
[2]Schwelm A,Fogelqvist J,Knaust A,et al.The Plasmodiophora brassicae genome reveals insights in its life cycle and ancestry of chitin synthases[J].Scientific Reports,2015(5):11153.
[3]战宗祥.甘蓝型油菜抗根肿病资源创新与PbBa8.1位点遗传转育[D].武汉:华中农业大学,2017.
[4]刘勇,黄小琴,柯绍英,等.四川主栽油菜品种根肿病抗性研究[J].中国油料作物学报,2009,31(1):90-09.
[5]王靖,黄云,胡晓玲,等.油菜根肿病症状、病原形态及产量损失研究[J].中国油料作物学报,2008,30(1):112-115.
[6]季海雯,任莉,陈坤荣,等.油菜根肿病病原主要生理小种和品种抗病性鉴定[J].中国油料作物学报,2013,35(3):301-306.
[7]严位中,杨家鸾,孙道旺,等.云南十字花科蔬菜根肿病发生规律及防治技术研究[J].石河子大学学报,2004,22:118-122.
[8]符明联,杨玉珠,李根泽,等.不同油菜品种对根肿病的抗性分析[J].华中农业大学学报,2011,30(4):443-447.
[9]Caroline D,Ian P.Integrated control of clubroot[J].Journal of Plant Growth Regulation,2009,28:289-303.
[10]Kageyama K,Asano T.Life cycle of Plasmodiophora brassicae[J].Journal of Plant Growth Regulation,2009,28:203-211.
[11]Saito M,Kubo N,Matsumoto S,et al.Fine mapping of the clubroot resistance gene Crr3 in Brassica rapa[J].Theoretical and Applied Genetics,2006,114:81-91.
[12]白亭亭,杨佩文,李向东,等.不同措施对十字花科作物根肿病控制效果研究[J].西南农业学报,2018,31(4):731-735.
[13]Hirai M.Genetic analysis of clubroot resistance in Brassica crops[J].Breeding Science,2006,56:223-229.
[14]刘勇,罗一帆,黄小琴,等.芸薹根肿菌生理小种鉴别方法研究进展[J].中国油料作物学报,2011,33(4):420-426.
[15]沈向群,聂凯,吴琼,等.大白菜根肿病主要生理小种种群分化鉴定初报[J].中国蔬菜,2009 (8):59-62.
[16]刘峰,张丽辉,姬广海.云南和西藏十字花科蔬菜根肿病菌生理小种鉴定[J].中国蔬菜,2013 (20):77-81.
[17]丁云花,简元才,余阳俊,等.我国8省市十字花科蔬菜根肿病菌生理小种的鉴定[J].中国蔬菜,2013(16):85-88.
[18]聂凯,沈向群,吴琼,等.大白菜抗根肿病雄性不育系转育效果的评价[J].江苏农业科学,2009(2):157-159.
[19]谭翀,岳艳玲.不同大白菜品种对根肿病的抗性鉴定[J].中国蔬菜,2013(8):91-94.
[20]李懋学,张赞平.作物染色体及其研究技术[M].北京:中国农业出版社,1996.
[21]杨佩文,李家瑞,杨勤忠,等.十字花科蔬菜根肿病菌休眠孢子的分离与检测[J].云南农业大学学报,2002,17(3):301-303.
[22]Siemens J,Nagel M,Ludwig-Müller J,et al.The interaction of Plasmodiophora brassicae and Arabidopsis thaliana:parameters for disease quantification and screening of mutant lines[J].J Phytopathol,2002,150:592-605.
[23]陈静,任雪松,宋洪元,等.4个不同地区十字花科根肿病菌生理小种鉴定及甘蓝新组合的抗性鉴定[J].西南大学学报(自然科学版),2016,38(1):67-72.
[24]黄小莉,任佐华,彭沙莎,等.湖南白菜品种根肿病抗性鉴定及评价[J].湖南农业大学学报(自然科学版),2014,40(1):43-47.
[2]Schwelm A,Fogelqvist J,Knaust A,et al.The Plasmodiophora brassicae genome reveals insights in its life cycle and ancestry of chitin synthases[J].Scientific Reports,2015(5):11153.
[3]战宗祥.甘蓝型油菜抗根肿病资源创新与PbBa8.1位点遗传转育[D].武汉:华中农业大学,2017.
[4]刘勇,黄小琴,柯绍英,等.四川主栽油菜品种根肿病抗性研究[J].中国油料作物学报,2009,31(1):90-09.
[5]王靖,黄云,胡晓玲,等.油菜根肿病症状、病原形态及产量损失研究[J].中国油料作物学报,2008,30(1):112-115.
[6]季海雯,任莉,陈坤荣,等.油菜根肿病病原主要生理小种和品种抗病性鉴定[J].中国油料作物学报,2013,35(3):301-306.
[7]严位中,杨家鸾,孙道旺,等.云南十字花科蔬菜根肿病发生规律及防治技术研究[J].石河子大学学报,2004,22:118-122.
[8]符明联,杨玉珠,李根泽,等.不同油菜品种对根肿病的抗性分析[J].华中农业大学学报,2011,30(4):443-447.
[9]Caroline D,Ian P.Integrated control of clubroot[J].Journal of Plant Growth Regulation,2009,28:289-303.
[10]Kageyama K,Asano T.Life cycle of Plasmodiophora brassicae[J].Journal of Plant Growth Regulation,2009,28:203-211.
[11]Saito M,Kubo N,Matsumoto S,et al.Fine mapping of the clubroot resistance gene Crr3 in Brassica rapa[J].Theoretical and Applied Genetics,2006,114:81-91.
[12]白亭亭,杨佩文,李向东,等.不同措施对十字花科作物根肿病控制效果研究[J].西南农业学报,2018,31(4):731-735.
[13]Hirai M.Genetic analysis of clubroot resistance in Brassica crops[J].Breeding Science,2006,56:223-229.
[14]刘勇,罗一帆,黄小琴,等.芸薹根肿菌生理小种鉴别方法研究进展[J].中国油料作物学报,2011,33(4):420-426.
[15]沈向群,聂凯,吴琼,等.大白菜根肿病主要生理小种种群分化鉴定初报[J].中国蔬菜,2009 (8):59-62.
[16]刘峰,张丽辉,姬广海.云南和西藏十字花科蔬菜根肿病菌生理小种鉴定[J].中国蔬菜,2013 (20):77-81.
[17]丁云花,简元才,余阳俊,等.我国8省市十字花科蔬菜根肿病菌生理小种的鉴定[J].中国蔬菜,2013(16):85-88.
[18]聂凯,沈向群,吴琼,等.大白菜抗根肿病雄性不育系转育效果的评价[J].江苏农业科学,2009(2):157-159.
[19]谭翀,岳艳玲.不同大白菜品种对根肿病的抗性鉴定[J].中国蔬菜,2013(8):91-94.
[20]李懋学,张赞平.作物染色体及其研究技术[M].北京:中国农业出版社,1996.
[21]杨佩文,李家瑞,杨勤忠,等.十字花科蔬菜根肿病菌休眠孢子的分离与检测[J].云南农业大学学报,2002,17(3):301-303.
[22]Siemens J,Nagel M,Ludwig-Müller J,et al.The interaction of Plasmodiophora brassicae and Arabidopsis thaliana:parameters for disease quantification and screening of mutant lines[J].J Phytopathol,2002,150:592-605.
[23]陈静,任雪松,宋洪元,等.4个不同地区十字花科根肿病菌生理小种鉴定及甘蓝新组合的抗性鉴定[J].西南大学学报(自然科学版),2016,38(1):67-72.
[24]黄小莉,任佐华,彭沙莎,等.湖南白菜品种根肿病抗性鉴定及评价[J].湖南农业大学学报(自然科学版),2014,40(1):43-47.
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