甜瓜霜霉病苗期抗病性鉴定方法建立及种质资源评价
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摘要
【目的】研究甜瓜霜霉病苗期人工接种鉴定的最佳方法,并对甜瓜资源进行苗期接种鉴定,为抗病育种提供抗源。【方法】采用不同接种方法,不同接种苗龄,不同接种浓度三因素三水平正交试验设计,建立甜瓜霜霉病苗期抗性鉴定方法,并对208份新疆哈密瓜资源和国内引进甜瓜资源进行霜霉病抗病性评价。【结果】甜瓜苗期接种霜霉病最好的方法是喷雾法,接种苗龄为2叶期,接种的最适浓度为5×10~3孢子/mL;筛选出免疫甜瓜霜霉病资源5份,高抗资源7份,中抗资源6份。【结论】甜瓜苗期接种霜霉病方法的确立以及抗霜霉病资源筛选为抗病育种奠定了基础。
【Objective】 To study the best method of artificial inoculation identification of melon downy mildew at seedling stage, and carry out inoculation identification of melon resources so as to provide resistance source for disease resistance breeding. 【Method】Three factors and three levels of orthogonal test design were used in different inoculation methods, different seedling ages, and different inoculation concentrations to establish a method for identifying the seedling resistance of melon downy mildew, and evaluate the disease resistance of 208 Xinjiang Hami melon resources and the introduced melon resources in China. 【Result】 The best method for melon seedling inoculation with downy mildew was spray method, the seedling age was 2 leaves, and the optimal concentration of inoculation was 5 ×10~3 spores/mL. Five resources, seven resources with high resistance and six resources with medium resistance were selected. 【Conclusion】The establishment of the method of inoculation against downy mildew at seedling stage and the selection of resources against downy mildew laid a foundation for disease resistance breeding.
【Objective】 To study the best method of artificial inoculation identification of melon downy mildew at seedling stage, and carry out inoculation identification of melon resources so as to provide resistance source for disease resistance breeding. 【Method】Three factors and three levels of orthogonal test design were used in different inoculation methods, different seedling ages, and different inoculation concentrations to establish a method for identifying the seedling resistance of melon downy mildew, and evaluate the disease resistance of 208 Xinjiang Hami melon resources and the introduced melon resources in China. 【Result】 The best method for melon seedling inoculation with downy mildew was spray method, the seedling age was 2 leaves, and the optimal concentration of inoculation was 5 ×10~3 spores/mL. Five resources, seven resources with high resistance and six resources with medium resistance were selected. 【Conclusion】The establishment of the method of inoculation against downy mildew at seedling stage and the selection of resources against downy mildew laid a foundation for disease resistance breeding.
引文
[1] Berkeley M S,Curtis A.Peronospora cubensis [J].Botanical Journal of the Linnean Society,1868,(10):363.
[2] Colucci S J ,Wehner T C ,Holmes G J .The downy mildew epidemic of 2004 and 2005 in the eastern United States [C]// Asheville North,Carolina,USA,17-21 September 2006.Organizing Committee,Cucurbitaceae,2006:403-411.
[3] 聂祥祥.甜瓜抗霜霉病无选择标记基因的转化和功能验证[D].乌鲁木齐:新疆大学,2014.NIE Xiang-xiang.(2014).Transformation and Verification for Marker Free Transgenic Melon with Resistance to Downy Mildew [D] Urumqi:Xinjiang University .(in Chinese)
[4] 贺玉花.甜瓜霜霉病抗性基因的SSR标记[D].北京.中国农业科学院,2014.HE Yu-hua.(2014).The SSR Markers Linked to Downy Mildew Resistant Gene in Melon (Cucumis melo)[D] Beijing:China Agricultural University.(in Chinese)
[5] Cohen Y,Cohen S,Eyal H,Thomas C E.(1984).Evaluating downy mildew resistance in Cucumis melo L[J].Cucurbit Genetics Cooperative Report,(7):38-40 .
[6] Cohen Y.(1977).The combined effects of temperature,leaf wetness,and inoculum concentration on infection of cucumbers with Pseudoperonospora cubensis [J].Canadian Journal of Botany,55(11):1,478-1,487.
[7] Perchepied L ,Bardin M ,Dogimont C ,et al.(2005).Relationship between loci conferring downy mildew and powdery mildew resistance in melon assessed by quantitative trait loci mapping [J].Phytopathology,95(5):556-565.
[8]张学军,郭丽霞,马新力,等.甜瓜种质资源抗霜霉病和蔓枯病苗期筛选与评价[J].中国蔬菜,2014,27(S1):38-40.ZHANG Xue-jun,GUO Li-xia,MA Xin-li,et al.Screening and evaluation of melon germplasm resources for resistance to downy mildew and vine blight at Seedling Stage[J].China Melon and Vegetable,2014,27(S1):38-40.
[9]翁祖信,冯东昕,李宝栋.黄瓜霜霉病抗病性鉴定技术研究初报[J].中国蔬菜,1991,(4):7-9.WENG Zu-xin,FENG Dong-xin,LI Bao-dong.(1991).Preliminary report on identification technology of cucumber downy mildew resistance[J].China Vegetables,(4):7-9.
[10]张学军,李寐华,张,等.甜瓜苗期复合接种多抗性方法研究[J].北方园艺,2013,(1):138-140.ZHANG Xue-jun,LI Mei-hua,ZHANG Yan,et al.( 2013).Study on the Method of Identification for Multiple Disease Resistance in Melon Seedling Stage[J].Northern Horticulture,(1):138-140.
[11] 张学军,宁雪飞,杨永,等.甜瓜PI390452霜霉病抗性基因的QTL定位[J].新疆农业科学,2016,53(12):2 157- 2 165.ZHANG Xue-jun,NING Xue-fei,YANG Yong,et al.(2016).QTL Mapping of Resistance Genes to Downy Mildew in Cucumis melo ssp.melo PI390452 [J].Xinjiang Agricultural Sciencws,53(12):2,157-2,165.(in Chinese)
[12] Criswell A D,Wehner T C,Klosinska U,Kozik E,Pitrat M.(2008).Use of sporulation and other leaf and vine traits for evaluation of resistance to downy mildew in cucumber [J].Proc.Cucurbitaceae 2008 EUCARPIA meeting:433-440.
[13] Epinat C ,Pitrat M .(1994).Inheritance of resistance to downy mildew (Pseudoperonospora cubensis) in muskmelon (Cucumis melo).II.Generation means analysis of 5 genitors [J].Agronomie,8(8):239-248.
[14] Epinat C,Pitrat M.(1994).Inheritance of resistance to downy mildew (Pseudoperonospora cubensis) in muskmelon (Cucumis melo).II.Generation means analysis of genitors[J].Agronomie,14(4):249-257.
[15] 吕淑珍,霍振荣,陈正式.黄瓜抗病遗传初报[J].天津农林科技,1990,(2):22-24.
[16] 龚振平,于拴仓,苏同兵,等.大白菜骨干自交系的苗期抗病性评价[J].植物遗传资源学报,2015,16(6):1 194-1 205.GONG Zhen-ping,YU Shuan-cang,SU Tong-bing,et al.(2015).Resistance to Main Diseases Evaluated at Seedling Stage in Core Set of Chinese Cabbage Inbred Lines [J].Journal of Plant Genetic Resources,16(6):1,194-1,205.(in Chinese)
[17] Cohen Y.(1977).The combined effects of temperature,leaf wetness,and inoculum concentration on infection of cucumbers with Pseudoperonospora cubensis [J].Canadian Journal of Botany,55(11):1,478-1,487.
[18] Epinat C,Pitrat M.(1994).Inheritance of resistance to downy mildew (Pseudoperonospora cubensis) in muskmelon (Cucumis melo).II.Generation means analysis of 5 genitors [J].AGRONOMIE -PARIS-.
[19] Neykov S,Dobrev D.(1988).Introduced Cucumber Cultivars Relatively Resistant to Pseudoperonospora Cubensis in Bulgaria [J].Acta Horticulturae,(220):115-120.
[20] Vliet G J A V,Meysing W D.(1974).Inheritance of resistance to Pseudoperonospora cubensisRost.In cucumber (Cucumis sativus L.) [J].Euphytica,23(2):251-255.
[21] Fanourakis N E,Simon P W.(1987) Analysis of genetic linkage in the cucumber [J].Journal of Heredity,78(4):238-242.
[22] Vliet G J A V,Meijsing W D.(1977).Relation in the inheritance of resistance to Pseudoperonospora cubensis Rost.and Sphaerotheca fuliginea Poll.in cucumber (Cucumis sativus L.)[J].Euphytica,26(3):793-796.
[23] Petrov L ,Boogert K ,Sheck L ,et al.(2000).Resistance to downy mildew,Pseudoperonospora cubensis,in cucumbers[J].Acta Horticulturae,510(510):203-209.
[24] 贺玉花,徐永阳,徐志红,等.甜瓜霜霉病抗性研究进展[J].果树学报,2014,31(2):324-334.HE Yu-hua,XU Yong-yang,XU Zhi-hong,et al.( 2014).Research progress of resistance to downy mildew in melon [J].Journal of Fruit Science,31(2):324-334.(in Chinese)
[2] Colucci S J ,Wehner T C ,Holmes G J .The downy mildew epidemic of 2004 and 2005 in the eastern United States [C]// Asheville North,Carolina,USA,17-21 September 2006.Organizing Committee,Cucurbitaceae,2006:403-411.
[3] 聂祥祥.甜瓜抗霜霉病无选择标记基因的转化和功能验证[D].乌鲁木齐:新疆大学,2014.NIE Xiang-xiang.(2014).Transformation and Verification for Marker Free Transgenic Melon with Resistance to Downy Mildew [D] Urumqi:Xinjiang University .(in Chinese)
[4] 贺玉花.甜瓜霜霉病抗性基因的SSR标记[D].北京.中国农业科学院,2014.HE Yu-hua.(2014).The SSR Markers Linked to Downy Mildew Resistant Gene in Melon (Cucumis melo)[D] Beijing:China Agricultural University.(in Chinese)
[5] Cohen Y,Cohen S,Eyal H,Thomas C E.(1984).Evaluating downy mildew resistance in Cucumis melo L[J].Cucurbit Genetics Cooperative Report,(7):38-40 .
[6] Cohen Y.(1977).The combined effects of temperature,leaf wetness,and inoculum concentration on infection of cucumbers with Pseudoperonospora cubensis [J].Canadian Journal of Botany,55(11):1,478-1,487.
[7] Perchepied L ,Bardin M ,Dogimont C ,et al.(2005).Relationship between loci conferring downy mildew and powdery mildew resistance in melon assessed by quantitative trait loci mapping [J].Phytopathology,95(5):556-565.
[8]张学军,郭丽霞,马新力,等.甜瓜种质资源抗霜霉病和蔓枯病苗期筛选与评价[J].中国蔬菜,2014,27(S1):38-40.ZHANG Xue-jun,GUO Li-xia,MA Xin-li,et al.Screening and evaluation of melon germplasm resources for resistance to downy mildew and vine blight at Seedling Stage[J].China Melon and Vegetable,2014,27(S1):38-40.
[9]翁祖信,冯东昕,李宝栋.黄瓜霜霉病抗病性鉴定技术研究初报[J].中国蔬菜,1991,(4):7-9.WENG Zu-xin,FENG Dong-xin,LI Bao-dong.(1991).Preliminary report on identification technology of cucumber downy mildew resistance[J].China Vegetables,(4):7-9.
[10]张学军,李寐华,张,等.甜瓜苗期复合接种多抗性方法研究[J].北方园艺,2013,(1):138-140.ZHANG Xue-jun,LI Mei-hua,ZHANG Yan,et al.( 2013).Study on the Method of Identification for Multiple Disease Resistance in Melon Seedling Stage[J].Northern Horticulture,(1):138-140.
[11] 张学军,宁雪飞,杨永,等.甜瓜PI390452霜霉病抗性基因的QTL定位[J].新疆农业科学,2016,53(12):2 157- 2 165.ZHANG Xue-jun,NING Xue-fei,YANG Yong,et al.(2016).QTL Mapping of Resistance Genes to Downy Mildew in Cucumis melo ssp.melo PI390452 [J].Xinjiang Agricultural Sciencws,53(12):2,157-2,165.(in Chinese)
[12] Criswell A D,Wehner T C,Klosinska U,Kozik E,Pitrat M.(2008).Use of sporulation and other leaf and vine traits for evaluation of resistance to downy mildew in cucumber [J].Proc.Cucurbitaceae 2008 EUCARPIA meeting:433-440.
[13] Epinat C ,Pitrat M .(1994).Inheritance of resistance to downy mildew (Pseudoperonospora cubensis) in muskmelon (Cucumis melo).II.Generation means analysis of 5 genitors [J].Agronomie,8(8):239-248.
[14] Epinat C,Pitrat M.(1994).Inheritance of resistance to downy mildew (Pseudoperonospora cubensis) in muskmelon (Cucumis melo).II.Generation means analysis of genitors[J].Agronomie,14(4):249-257.
[15] 吕淑珍,霍振荣,陈正式.黄瓜抗病遗传初报[J].天津农林科技,1990,(2):22-24.
[16] 龚振平,于拴仓,苏同兵,等.大白菜骨干自交系的苗期抗病性评价[J].植物遗传资源学报,2015,16(6):1 194-1 205.GONG Zhen-ping,YU Shuan-cang,SU Tong-bing,et al.(2015).Resistance to Main Diseases Evaluated at Seedling Stage in Core Set of Chinese Cabbage Inbred Lines [J].Journal of Plant Genetic Resources,16(6):1,194-1,205.(in Chinese)
[17] Cohen Y.(1977).The combined effects of temperature,leaf wetness,and inoculum concentration on infection of cucumbers with Pseudoperonospora cubensis [J].Canadian Journal of Botany,55(11):1,478-1,487.
[18] Epinat C,Pitrat M.(1994).Inheritance of resistance to downy mildew (Pseudoperonospora cubensis) in muskmelon (Cucumis melo).II.Generation means analysis of 5 genitors [J].AGRONOMIE -PARIS-.
[19] Neykov S,Dobrev D.(1988).Introduced Cucumber Cultivars Relatively Resistant to Pseudoperonospora Cubensis in Bulgaria [J].Acta Horticulturae,(220):115-120.
[20] Vliet G J A V,Meysing W D.(1974).Inheritance of resistance to Pseudoperonospora cubensisRost.In cucumber (Cucumis sativus L.) [J].Euphytica,23(2):251-255.
[21] Fanourakis N E,Simon P W.(1987) Analysis of genetic linkage in the cucumber [J].Journal of Heredity,78(4):238-242.
[22] Vliet G J A V,Meijsing W D.(1977).Relation in the inheritance of resistance to Pseudoperonospora cubensis Rost.and Sphaerotheca fuliginea Poll.in cucumber (Cucumis sativus L.)[J].Euphytica,26(3):793-796.
[23] Petrov L ,Boogert K ,Sheck L ,et al.(2000).Resistance to downy mildew,Pseudoperonospora cubensis,in cucumbers[J].Acta Horticulturae,510(510):203-209.
[24] 贺玉花,徐永阳,徐志红,等.甜瓜霜霉病抗性研究进展[J].果树学报,2014,31(2):324-334.HE Yu-hua,XU Yong-yang,XU Zhi-hong,et al.( 2014).Research progress of resistance to downy mildew in melon [J].Journal of Fruit Science,31(2):324-334.(in Chinese)