SSR标记和苗期人工接种鉴定黄瓜抗黑星病种质资源
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摘要
为了获得抗黑星病黄瓜种质,为分子设计育种提供抗源,利用与黄瓜抗黑星病基因紧密连锁的SSR标记CSWCTT02D对102份黄瓜种质资源进行基因型分析。结果表明,黄瓜种质间抗病基因的标记基因型存在遗传变异性,明确了102份种质抗黑星病基因的标记基因型,3份种质存在抗病标记(2.94%),1份种质为杂合型(0.98%),98份种质不存在抗病标记(96.08%);苗期人工接种鉴定有高度抗病种质4份(3.92%),高度感病种质98份(96.08%)。SSR分子检测结果与苗期人工接种基本一致,有2份材料与人工接种鉴定结果不符。‘南9436’(华南型)接种为感病,标记为抗病;‘K92’(华南型)接种为抗病,标记为感病,符合率达98.04%。抗病材料选择应以人工接种结果为依据,因此初步鉴定出4份抗黑星病种质,分别为日本类型‘Q6’和‘NINIA’、华南型‘南9427’和‘K92’,病情指数均为0。该研究为华北型黄瓜抗黑星病品种的遗传改良奠定了技术与种质基础。
In order to obtain cucumber germplasm resistant to scab, provide resistance sources for molecular design breeding, and finally breeding resistant varieties to control cucumber scab. In this study,SSR marker CSWCTT02D linked to scab-resistance gene was used to identify the marker genotypes of 102 cucumber germplasm resources. The results showed genetic variation of marker genotypes exists among different germplasm and within same germplasm as well. The marker genotypes of 102 germplasm were confirmed, 3 germplasm had disease resistance marker(2.94%), one germplasm was heterozygous(0.98%), and 98 germplasm had no disease resistance marker(96.08%). There were 4 highly resistant germplasm(3.92%) and 98 highly susceptible germplasm(96.08%) identified by artificial inoculation at seedling stage. The results of SSR molecular detection were basically consistent with that of artificial inoculation at seedling stage, and 2 materials did not match the identification results of artificial inoculation. South'9436'was inoculated as susceptible and marked as resistant,'K92' was inoculated with disease resistance and marked as susceptible, with a coincidence rate of 98.04%. The selection of disease-resistant materials should be based on the results of artificial inoculation, so 4 resistant cucumber germplasm were preliminarily identified, namely'Q6','Ninia','Nan9427' and 'K92', with disease indices of 0. This study laid technical and germplasm foundation for improving scab-resistance cucumber varieties of north China cucumber.
In order to obtain cucumber germplasm resistant to scab, provide resistance sources for molecular design breeding, and finally breeding resistant varieties to control cucumber scab. In this study,SSR marker CSWCTT02D linked to scab-resistance gene was used to identify the marker genotypes of 102 cucumber germplasm resources. The results showed genetic variation of marker genotypes exists among different germplasm and within same germplasm as well. The marker genotypes of 102 germplasm were confirmed, 3 germplasm had disease resistance marker(2.94%), one germplasm was heterozygous(0.98%), and 98 germplasm had no disease resistance marker(96.08%). There were 4 highly resistant germplasm(3.92%) and 98 highly susceptible germplasm(96.08%) identified by artificial inoculation at seedling stage. The results of SSR molecular detection were basically consistent with that of artificial inoculation at seedling stage, and 2 materials did not match the identification results of artificial inoculation. South'9436'was inoculated as susceptible and marked as resistant,'K92' was inoculated with disease resistance and marked as susceptible, with a coincidence rate of 98.04%. The selection of disease-resistant materials should be based on the results of artificial inoculation, so 4 resistant cucumber germplasm were preliminarily identified, namely'Q6','Ninia','Nan9427' and 'K92', with disease indices of 0. This study laid technical and germplasm foundation for improving scab-resistance cucumber varieties of north China cucumber.
引文
[1]易齐,王蔚,王传英.黄瓜黑星病及其蔓延危害现状[J].植物保护,1987(6):40-41.
[2]王进扬,卢香春,王敏.黄瓜黑星病调查简报[J].中国蔬菜,1988(2):42-43.
[3]李明远,易齐.黄瓜黑星病在我国流行的现状[J].长江蔬菜,1989(3):26-27.
[4]BAILEY R M,BURGESS I M.Breeding Cucumbers resistant to scab.[J].Proceedings American Society for Horticultural Science,1934,32:474-476.
[5]张桂华,韩毅科,孙小红,等.与黄瓜抗黄瓜黑星病基因连锁的分子标记研究[J].中国农业科学,2006,39(11):2250-2254.
[6]王惠哲,李淑菊,管炜.与黄瓜抗黑星病相关基因紧密连锁的SSR标记[J].分子植物育种,2009,7(3):550-554.
[7]KANG H X,WENG Y Q,YANG Y H,et al.Fine genetic mapping localizes cucumber scab resistance gene Ccu into an Rgene cluster[J].Theoretical Applied Genetics,2011,122:795-803.
[8]王惠哲,李淑菊,杨瑞环,等.黄瓜枯萎病、炭疽病、褐斑病、黑星病苗期抗源材料筛选试验[J].农业科技通讯,2013(8):118-121.
[9]王惠哲,管炜,杨瑞环,等.黄瓜种质资源抗黑星病鉴定与评价[J].农业科技通讯,2013(12):135-137.
[10]ABUL-HAYJA Z,WILLIAMS P H.Inheritance of two seedling markers in cucumber[J].HortScience,1976,11(2):145.
[11]李光,李淑菊,王惠哲.兼抗黄瓜枯萎病、角斑病和黑星病育种材料苗期筛选[J].天津农业科学,2007,13(3):23-25.
[12]王艳飞,李平,王惠哲.黄瓜枯萎病、白粉病、霜霉病和黑星病苗期多抗性鉴定方法的评价[J].中国蔬菜,2007(3):27-28.
[13]王惠哲,李淑菊.黄瓜枯萎病-白粉病-褐斑病-黑星病多抗性鉴定技术研究[J].中国瓜菜,2010,23(5):6-8.
[14]李全辉,李锡香,王海平,等.黄瓜抗黑星病相关基因的差异表达分析[J].植物遗传资源学报,2013,14(3):501-506.
[15]毛爱军,张峰,张丽蓉,等.黄瓜品系WIS2757对黄瓜枯萎病生理小种4和黑星病的抗性遗传与连锁分析[J].中国农业科学,2008,41(10):3382-3388.
[16]康厚祥.黄瓜抗黑星病基因Ccu的精细定位与图位克隆及黄瓜与黑星菌互作的表达谱分析[D].北京:中国农业科学院,2011.
[17]周倩.基于基因组测序的黄瓜高密度遗传图谱构建和果皮浅绿突变体基因定位[D].北京:中国农业科学院,2015.
[2]王进扬,卢香春,王敏.黄瓜黑星病调查简报[J].中国蔬菜,1988(2):42-43.
[3]李明远,易齐.黄瓜黑星病在我国流行的现状[J].长江蔬菜,1989(3):26-27.
[4]BAILEY R M,BURGESS I M.Breeding Cucumbers resistant to scab.[J].Proceedings American Society for Horticultural Science,1934,32:474-476.
[5]张桂华,韩毅科,孙小红,等.与黄瓜抗黄瓜黑星病基因连锁的分子标记研究[J].中国农业科学,2006,39(11):2250-2254.
[6]王惠哲,李淑菊,管炜.与黄瓜抗黑星病相关基因紧密连锁的SSR标记[J].分子植物育种,2009,7(3):550-554.
[7]KANG H X,WENG Y Q,YANG Y H,et al.Fine genetic mapping localizes cucumber scab resistance gene Ccu into an Rgene cluster[J].Theoretical Applied Genetics,2011,122:795-803.
[8]王惠哲,李淑菊,杨瑞环,等.黄瓜枯萎病、炭疽病、褐斑病、黑星病苗期抗源材料筛选试验[J].农业科技通讯,2013(8):118-121.
[9]王惠哲,管炜,杨瑞环,等.黄瓜种质资源抗黑星病鉴定与评价[J].农业科技通讯,2013(12):135-137.
[10]ABUL-HAYJA Z,WILLIAMS P H.Inheritance of two seedling markers in cucumber[J].HortScience,1976,11(2):145.
[11]李光,李淑菊,王惠哲.兼抗黄瓜枯萎病、角斑病和黑星病育种材料苗期筛选[J].天津农业科学,2007,13(3):23-25.
[12]王艳飞,李平,王惠哲.黄瓜枯萎病、白粉病、霜霉病和黑星病苗期多抗性鉴定方法的评价[J].中国蔬菜,2007(3):27-28.
[13]王惠哲,李淑菊.黄瓜枯萎病-白粉病-褐斑病-黑星病多抗性鉴定技术研究[J].中国瓜菜,2010,23(5):6-8.
[14]李全辉,李锡香,王海平,等.黄瓜抗黑星病相关基因的差异表达分析[J].植物遗传资源学报,2013,14(3):501-506.
[15]毛爱军,张峰,张丽蓉,等.黄瓜品系WIS2757对黄瓜枯萎病生理小种4和黑星病的抗性遗传与连锁分析[J].中国农业科学,2008,41(10):3382-3388.
[16]康厚祥.黄瓜抗黑星病基因Ccu的精细定位与图位克隆及黄瓜与黑星菌互作的表达谱分析[D].北京:中国农业科学院,2011.
[17]周倩.基于基因组测序的黄瓜高密度遗传图谱构建和果皮浅绿突变体基因定位[D].北京:中国农业科学院,2015.