番茄叶霉病抗病基因Cf_（11）、Cf_（19）的分子标记研究
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摘要
本研究首先分析鉴定了东北三省番茄主要种植区番茄叶霉病生理小种的种群结构。并在番茄叶霉病抗性遗传分析的基础上,利用AFLP和SSR技术找到了与番茄抗叶霉病基因Cf_(11)和Cf_(19)连锁的AFLP标记和SSR标记,同时还利用的到的AFLP标记对番茄种质资源进行了分子鉴定和人工接种鉴定。主要研究结果如下:
1.对黑龙江、吉林和辽宁三省的番茄叶霉病菌生理小种的分析鉴定结果为:采集到的33个菌株主要为生理小种1.2、2.3、1.2.3、1.2.4和1.2.3.4。其中以小种1.2.3.4为主,占调查总数的57.75%,其次是生理小种1.2.3占调查总数的20.1%。
2.本试验分析了组合HN19(感病品种)×HN43(抗病品种)的杂交后代对番茄叶霉病菌1.2.3.4生理小种的抗病反应。经鉴定表明:两个亲本间抗、感差异明显,父本HN43均表现抗病,母本HN19均表现感病;F2群体抗病与感病株数的分离比为:3.08:1.00,符合3:1的分离比例。分析结果表明:父本HN43对番茄叶霉病菌1.2.3.4生理小种的抗性是由单基因控制的显性性状。
3.本试验分析了组合HN16(感病品种)×HN42(抗病品种)的杂交后代对番茄叶霉病菌1.2.3.4生理小种的抗病反应。经鉴定表明:两个亲本间抗、感差异明显,父本HN42均表现抗病,母本HN16均表现感病;F2群体抗病与感病株数的分离比为:3.17:1.00,符合3:1的分离比例。分析结果表明:父本HN42对番茄叶霉病菌1.2.3.4生理小种的抗性是由单基因控制的显性性状。
4.筛选到6个与抗番茄叶霉病基因Cf_(19)连锁的标记。其中5个AFLP标记E38M61-2、E66M84-4、E66M44-1、E77M50-1和E77M50-2与Cf_(19)的遗传距离分别为9.7 cM、10.7cM、12.75 cM、13.45 cM和14.4 cM:1个SSR标记SSR318与Cf_(19)的遗传距离是12.5 cM。
5.筛选到5个与抗番茄叶霉病基因Cf_(11)连锁的标记。其中4个AFLP标记E54M37-G、E62M1-A、E85M79-D、和E62M59-G与Cf_(11)遗传距离分别为10.1 cM、12.3cM、14.5 cM和18.8cM;1个SSR标记SSR46与Cf_(11)遗传距离是13.7cM。
6.对95份番茄种质资源进行了叶霉病生理小种1.2.3.4的抗性鉴定,人工接种鉴定共得到15份抗病资源,应用2个AFLP标记鉴定得到3份含有Cf_(11)基因的抗病材料和2份含有Cf_(11)基因的抗病材料。分子鉴定的结果与人工接种鉴定的结果高度吻合。在番茄抗叶霉病育种的工作中应考虑如何对这些抗病资源进行有效的利有,从而培育出多抗性品种,防止品种的抗性快速丧失。
The study collected and identified tomato leaf mold pathogen strains in main tomato growth area of northeast three provinces.Based on the genetic analysis of resistance to Cladosporium fulvum,AFLP and SSR markers that linked to the resistance gene Cf_(11) and Cf_(19) were identified.At the same time,tomato germplasm resistant to Cladosporium fulvum were screened with inculation and mocular markers on AFLP.
1.The article analysed and identified leaf mold pathogen strains differentiation.The results indicated that physiological race 1.2,race2.3,race 1.2.3,race1.2.4 and race1.2.3.4 were found.The race 1.2.3.4 and race 1.2.4 was dominant race.Race 1.2.3.4 occupied 57.75%,and race 1.2.3 occupied 20.1%.
2.Resistance to race 1.2.3.4 of Cladosporium fulvum was found to due to a single dominant gene through analyzing resistance behavior of the cross HN19(susceptible)×NHN43 (resistable).The results indicated that the diffence of two parents resistanc and susceptibility is obvious,the male parent HN43 shows resistance and the female parent HN19 shows susceptibility.The ratio of number of resistance and susceptibility on F2 is 3.08:1.00,which accord with the ratio of 3:1,which accord with the ratio of 1:1.The results shows resistance to Cladosporiumfulvum is dominated by one dominant gene.
3.Resistance to race 1.2.3.4 of Cladosporiumfulvum was found to due to a single dominant gene through analyzing resistance behavior of the cross HN16(susceptible)×NHN42 (resistable).The results indicated that the diffence of two parents resistanc and susceptibility is obvious,the male parent HN42 shows resistance and the female parent HN16 shows susceptibility.The ratio of number of resistance and susceptibility on F2 is 3.17:1.00,which accord with the ratio of 3:1,which accord with the ratio of 1:1.The results shows resistance to Cladosporium fulvum is dominated by one dominant gene.
4.Five AFLP markers and one SSR markers linked to the resistant gene Cf_(19) were identified. The genetic distance of five AFLP markers on E38M61-2,E66M84-4,E66M44-1,E77M50-1 and 77M50-2 are 9.7 cM、10.7cM、12.75 cM、13.45 cMand14.4 cM differently.The genetic distance of one SSR markers on SSR318 is 12.5 cM.
5.Four AFLP markers and one SSR markers linked to the resistant gene Cf_(11) were identified. The genetic distance of four AFLP markers on E54M37-G,E62M1-A,E85M79-D and E62M59-G are 10.1 cM,12.3cM,14.5 cM and 18.8cM differently.The genetic distance of one SSR markers on SSR46 is 13.7 cM.
6.Using AFLP marke to screen the tomato germplasms,at the same time identifying the resistance with race 1.2.3.4 in the greenhouse.There are 15 of 95 cultivars and lines were resistant tomato germplasm resoures.Among them 3 have Cf_(19) gene,and 2 have Cf_(11) gene.This the AFLP marker improved is credible.We should think that how to make use of them to breed resisting variety and to defend the loss of resistance.
1.对黑龙江、吉林和辽宁三省的番茄叶霉病菌生理小种的分析鉴定结果为:采集到的33个菌株主要为生理小种1.2、2.3、1.2.3、1.2.4和1.2.3.4。其中以小种1.2.3.4为主,占调查总数的57.75%,其次是生理小种1.2.3占调查总数的20.1%。
2.本试验分析了组合HN19(感病品种)×HN43(抗病品种)的杂交后代对番茄叶霉病菌1.2.3.4生理小种的抗病反应。经鉴定表明:两个亲本间抗、感差异明显,父本HN43均表现抗病,母本HN19均表现感病;F2群体抗病与感病株数的分离比为:3.08:1.00,符合3:1的分离比例。分析结果表明:父本HN43对番茄叶霉病菌1.2.3.4生理小种的抗性是由单基因控制的显性性状。
3.本试验分析了组合HN16(感病品种)×HN42(抗病品种)的杂交后代对番茄叶霉病菌1.2.3.4生理小种的抗病反应。经鉴定表明:两个亲本间抗、感差异明显,父本HN42均表现抗病,母本HN16均表现感病;F2群体抗病与感病株数的分离比为:3.17:1.00,符合3:1的分离比例。分析结果表明:父本HN42对番茄叶霉病菌1.2.3.4生理小种的抗性是由单基因控制的显性性状。
4.筛选到6个与抗番茄叶霉病基因Cf_(19)连锁的标记。其中5个AFLP标记E38M61-2、E66M84-4、E66M44-1、E77M50-1和E77M50-2与Cf_(19)的遗传距离分别为9.7 cM、10.7cM、12.75 cM、13.45 cM和14.4 cM:1个SSR标记SSR318与Cf_(19)的遗传距离是12.5 cM。
5.筛选到5个与抗番茄叶霉病基因Cf_(11)连锁的标记。其中4个AFLP标记E54M37-G、E62M1-A、E85M79-D、和E62M59-G与Cf_(11)遗传距离分别为10.1 cM、12.3cM、14.5 cM和18.8cM;1个SSR标记SSR46与Cf_(11)遗传距离是13.7cM。
6.对95份番茄种质资源进行了叶霉病生理小种1.2.3.4的抗性鉴定,人工接种鉴定共得到15份抗病资源,应用2个AFLP标记鉴定得到3份含有Cf_(11)基因的抗病材料和2份含有Cf_(11)基因的抗病材料。分子鉴定的结果与人工接种鉴定的结果高度吻合。在番茄抗叶霉病育种的工作中应考虑如何对这些抗病资源进行有效的利有,从而培育出多抗性品种,防止品种的抗性快速丧失。
The study collected and identified tomato leaf mold pathogen strains in main tomato growth area of northeast three provinces.Based on the genetic analysis of resistance to Cladosporium fulvum,AFLP and SSR markers that linked to the resistance gene Cf_(11) and Cf_(19) were identified.At the same time,tomato germplasm resistant to Cladosporium fulvum were screened with inculation and mocular markers on AFLP.
1.The article analysed and identified leaf mold pathogen strains differentiation.The results indicated that physiological race 1.2,race2.3,race 1.2.3,race1.2.4 and race1.2.3.4 were found.The race 1.2.3.4 and race 1.2.4 was dominant race.Race 1.2.3.4 occupied 57.75%,and race 1.2.3 occupied 20.1%.
2.Resistance to race 1.2.3.4 of Cladosporium fulvum was found to due to a single dominant gene through analyzing resistance behavior of the cross HN19(susceptible)×NHN43 (resistable).The results indicated that the diffence of two parents resistanc and susceptibility is obvious,the male parent HN43 shows resistance and the female parent HN19 shows susceptibility.The ratio of number of resistance and susceptibility on F2 is 3.08:1.00,which accord with the ratio of 3:1,which accord with the ratio of 1:1.The results shows resistance to Cladosporiumfulvum is dominated by one dominant gene.
3.Resistance to race 1.2.3.4 of Cladosporiumfulvum was found to due to a single dominant gene through analyzing resistance behavior of the cross HN16(susceptible)×NHN42 (resistable).The results indicated that the diffence of two parents resistanc and susceptibility is obvious,the male parent HN42 shows resistance and the female parent HN16 shows susceptibility.The ratio of number of resistance and susceptibility on F2 is 3.17:1.00,which accord with the ratio of 3:1,which accord with the ratio of 1:1.The results shows resistance to Cladosporium fulvum is dominated by one dominant gene.
4.Five AFLP markers and one SSR markers linked to the resistant gene Cf_(19) were identified. The genetic distance of five AFLP markers on E38M61-2,E66M84-4,E66M44-1,E77M50-1 and 77M50-2 are 9.7 cM、10.7cM、12.75 cM、13.45 cMand14.4 cM differently.The genetic distance of one SSR markers on SSR318 is 12.5 cM.
5.Four AFLP markers and one SSR markers linked to the resistant gene Cf_(11) were identified. The genetic distance of four AFLP markers on E54M37-G,E62M1-A,E85M79-D and E62M59-G are 10.1 cM,12.3cM,14.5 cM and 18.8cM differently.The genetic distance of one SSR markers on SSR46 is 13.7 cM.
6.Using AFLP marke to screen the tomato germplasms,at the same time identifying the resistance with race 1.2.3.4 in the greenhouse.There are 15 of 95 cultivars and lines were resistant tomato germplasm resoures.Among them 3 have Cf_(19) gene,and 2 have Cf_(11) gene.This the AFLP marker improved is credible.We should think that how to make use of them to breed resisting variety and to defend the loss of resistance.
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