分子标记辅助选择Pi9与Pigm基因培育抗稻瘟病籼稻新品系
发布时间：2018-11-09 18:00
【摘要】：水稻是全球半数以上人口的主要口粮,特别是在一些东南亚国家。稻瘟病是水稻生产中最严重的病害之一,普遍分布于各个水稻产区,每年因此造成大量粮食损失。本论文研究分别以含稻瘟病广谱抗性基因Pi9的水稻品系75-1-127和含稻瘟病广谱抗性基因Pigm的水稻品种谷梅4号作供体亲本,以35份籼稻品种或品系作为受体亲本及轮回亲本,设计并利用Pi9基因内功能标记和Pigm基因紧密连锁标记开展分子标记辅助选择(Molecular Marker-assisted Selection, MAS)育种实践,定向改良受体亲本的稻瘟病抗性,获得了如下研究结果：1.根据已克隆的广谱持久抗瘟基因Pi9的基因序列信息设计基因内特异功能标记Clon2-1,该标记为共显性DNA标记,用它在35份水稻受体亲本和Pi9基因供体亲本75-1-127进行基因型分析,结果表明在双亲间多态率为97.1%。2.根据广谱持久抗瘟基因Pigm的基因精细定位信息设计了紧密连锁DNA标记T9E3,基因型分析结果表明该标记为共显性标记,在供体亲本谷梅4号与35份受体亲本间的多态率为100%。3.田间病圃抗性鉴定表明,除Pi9基因供体75-1-127与Pigm基因供体谷梅4号外,35份籼稻受体亲本的稻瘟病抗性均不强、抗谱不广,都表现不同程度的感病；室内接种抗性鉴定与抗菌谱分析结果表明,75-1-127与谷梅4号对17份来自不同国家或地区的稻瘟菌小种(菌株)的抗性频率分别达到88.2%与94.1%,供试的25份受体亲本的抗性频率在16.7%-82.3%之间,大部分受体亲本的稻瘟病抗性急需改良。4.对回交群体中随机取样个体的基因型分析和田间病圃对应表型鉴定结果,证明分子标记Clon2-1和T9E3对稻瘟病抗性表型选择效率均可达到100%,说明可以用这两个标记有效开展MAS育种实践。5.通过分子标记辅助选择和长沙三亚两地连续回交育种,定向改良35份籼稻受体亲本的稻瘟病抗性,获得了11个受体亲本的48份含Pi9基因(位点)的高抗稻瘟病水稻新品系,并用其中4个抗病纯系配制了4个杂交种,同时获得了其他24份受体亲本的不同世代回交群体；获得了10个受体亲本的含pigm基因(位点)的BC6F1高代回交群体,以及25个受体亲木的不同世代回交群体。这些工作为进一步培育广谱高抗稻瘟病水稻新品种(组合)奠定了基础。
[Abstract]:Rice is the staple food for more than half of the world's population, especially in some Southeast Asian countries. Rice blast is one of the most serious diseases in rice production. In this paper, rice lines 75-1-127 containing broad-spectrum resistance gene Pi9 and rice variety Gumei 4 containing Pigm gene were used as donor parents, respectively. 35 indica rice varieties or lines were used as recipient parents and recurrent parents to design and utilize Pi9 gene internal function marker and Pigm gene close linkage marker to carry out molecular marker-assisted selection (Molecular Marker-assisted Selection, MAS) breeding practice). The rice blast resistance of directed modified receptor parents was studied as follows: 1. According to the gene sequence information of the cloned broad-spectrum persistent blast resistance gene Pi9, a special gene marker Clon2-1, was designed. The marker was codominant DNA marker. The genotypes of 35 rice recipient parents and 75-1-127 donor parents of Pi9 gene were analyzed. The results showed that the polymorphism rate between parents was 97.1.2. A tightly linked DNA marker T9E3 was designed based on the gene fine mapping information of broad-spectrum persistent blast resistance gene Pigm. The results of genotype analysis showed that the marker was codominant. The polymorphic rate between donor parent Gu Mei 4 and 35 recipient parents was 100. 3. The identification of field disease resistance showed that, except for Pi9 gene donor 75-1-127 and Pigm gene donor Gamei 4, 35 indica rice receptor parents had low resistance to rice blast, and their resistance spectrum was not wide, and all of them were susceptible to different degrees of disease. The results of indoor inoculation resistance identification and antimicrobial spectrum analysis showed that the resistance frequency of 75-1-127 and Gumei No. 4 to 17 strains (strains) of Magnaporthe grisea from different countries or regions were 88.2% and 94.1%, respectively. The resistance frequency of 25 recipient parents ranged from 16.7- 82.3%, and the resistance of most recipient parents to rice blast was in urgent need of improvement. 4. Genotypic analysis of randomly sampled individuals in backcross population and identification of phenotypic characteristics in field disease beds showed that the selection efficiency of molecular markers Clon2-1 and T9E3 for blast resistance could reach 100%. These two markers can be used to develop MAS breeding practice effectively. 5. 5. By means of molecular marker-assisted selection and continuous backcross breeding in Sanya, Changsha, 35 indica rice receptor parents were directed to improve their rice blast resistance. 48 new rice lines with high resistance to rice blast with Pi9 gene (locus) were obtained from 11 recipient parents. Four hybrids were prepared with 4 resistant pure lines, and the backcross populations of other 24 recipient parents in different generations were obtained. The high generation backcross populations of BC6F1 containing pigm gene (loci) of 10 recipient parents and 25 backcross populations of different generations of recipient parents were obtained. These work have laid a foundation for further breeding new rice varieties (combinations) with wide spectrum and high blast resistance.
【学位授予单位】：湖南农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S435.111.41
[Abstract]:Rice is the staple food for more than half of the world's population, especially in some Southeast Asian countries. Rice blast is one of the most serious diseases in rice production. In this paper, rice lines 75-1-127 containing broad-spectrum resistance gene Pi9 and rice variety Gumei 4 containing Pigm gene were used as donor parents, respectively. 35 indica rice varieties or lines were used as recipient parents and recurrent parents to design and utilize Pi9 gene internal function marker and Pigm gene close linkage marker to carry out molecular marker-assisted selection (Molecular Marker-assisted Selection, MAS) breeding practice). The rice blast resistance of directed modified receptor parents was studied as follows: 1. According to the gene sequence information of the cloned broad-spectrum persistent blast resistance gene Pi9, a special gene marker Clon2-1, was designed. The marker was codominant DNA marker. The genotypes of 35 rice recipient parents and 75-1-127 donor parents of Pi9 gene were analyzed. The results showed that the polymorphism rate between parents was 97.1.2. A tightly linked DNA marker T9E3 was designed based on the gene fine mapping information of broad-spectrum persistent blast resistance gene Pigm. The results of genotype analysis showed that the marker was codominant. The polymorphic rate between donor parent Gu Mei 4 and 35 recipient parents was 100. 3. The identification of field disease resistance showed that, except for Pi9 gene donor 75-1-127 and Pigm gene donor Gamei 4, 35 indica rice receptor parents had low resistance to rice blast, and their resistance spectrum was not wide, and all of them were susceptible to different degrees of disease. The results of indoor inoculation resistance identification and antimicrobial spectrum analysis showed that the resistance frequency of 75-1-127 and Gumei No. 4 to 17 strains (strains) of Magnaporthe grisea from different countries or regions were 88.2% and 94.1%, respectively. The resistance frequency of 25 recipient parents ranged from 16.7- 82.3%, and the resistance of most recipient parents to rice blast was in urgent need of improvement. 4. Genotypic analysis of randomly sampled individuals in backcross population and identification of phenotypic characteristics in field disease beds showed that the selection efficiency of molecular markers Clon2-1 and T9E3 for blast resistance could reach 100%. These two markers can be used to develop MAS breeding practice effectively. 5. 5. By means of molecular marker-assisted selection and continuous backcross breeding in Sanya, Changsha, 35 indica rice receptor parents were directed to improve their rice blast resistance. 48 new rice lines with high resistance to rice blast with Pi9 gene (locus) were obtained from 11 recipient parents. Four hybrids were prepared with 4 resistant pure lines, and the backcross populations of other 24 recipient parents in different generations were obtained. The high generation backcross populations of BC6F1 containing pigm gene (loci) of 10 recipient parents and 25 backcross populations of different generations of recipient parents were obtained. These work have laid a foundation for further breeding new rice varieties (combinations) with wide spectrum and high blast resistance.
【学位授予单位】：湖南农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S435.111.41
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