云南糯稻遗传多样性研究
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摘要
糯稻(Oryza sativa var. glutinosae)是栽培稻的粘性变种,具有很高的食用价值和经济价值。云南省是亚洲栽培稻的起源和多样性分布中心之一,也是中国糯稻资源最为丰富的省份之一。为了进一步了解云南糯稻品种的遗传多样性丰度、分布、品种间亲缘关系、籼粳分化以及抗病性情况,本研究以云南糯稻品种为材料,利用SSR分子标记进行了遗传多样性分析;利用籼粳特异性InDel分子标记鉴定籼粳分化;进行了对稻瘟病的抗性评价。取得的主要研究结果如下:
1.选用均匀分布于水稻12条染色体的24对SSR(simple sequence repeat)引物,对来自云南13个地州36个县区的150份糯稻品种1500个单株的遗传多样性研究表明:24个SSR位点共检测到158个等位基因,每个位点平均多态信息含量为0.7262,均为中度或高度多态位点。总群体平均有效等位基因数为4.5506,平均香农指数为1.5928,平均期望杂合度为0.7642。云南糯稻总体遗传多样性水平较高。分子变异分析表明糯稻的遗传变异主要存在于品种间(86%),品种内的遗传变异占14%。保护糯稻遗传多样性需要保护较多的品种。滇南、滇东南和滇西南糯稻品种的遗传多样性显著高于滇中、滇东北和滇西北糯稻品种的遗传多样性。滇南、滇东南和滇西南是目前云南省糯稻遗传多样性中心,是保护糯稻资源时应重点关注的区域。聚类分析显示Nei遗传相似系数为0.19-0.99时150份云南糯稻品种存在丰富的遗传变异,但是不能区分地理分组。
2.用28对籼粳特异性插入缺失(insertion deletion, InDel)引物和24对简单重复序列( SSR )分析云南60份糯稻品种的籼粳分化和遗传变异,结果表明有24份糯稻品种为“典型籼型”,6份为“籼型”,2份为“粳型”,28份为“典型粳型”。聚类分析、主成分分析和基于模型的结构分析均表明糯稻籼粳分化明显,籼糯和粳糯属于两个独立的类群。遗传变异分析表明籼糯、粳糯的遗传多样性均较丰富,且SSR多态性差异不显著。分子变异分析表明糯稻的遗传变异主要来自亚种内(占总变异的74%),亚种间遗传变异占26%,但分化极显著。
3.采用温室人工接种方法用云南省的8个稻瘟病菌优势小种对100份云南糯稻品种进行抗稻瘟病鉴定,结果表明,中感以上品种和中感品种分别占到64%和34%,只有2份糯稻品种表现为感病,目前栽培糯稻品种较抗病。
The glutinous rice (Oryza sativa var. glutinosae) is the glutinosity type of rice variety that has high edible and economic value. Yunnan province is one of the richest Provinces on glutinous rice resource in China, as well as one of the origins and diversity centers of rice (Oryza sativa). In this report, the genetic diversity and structure of Yunnan glutinous rice varieties was analyzed by simple sequence repeat (SSR) markers. The Indica-Japonica differetiation was analyzed by insertion/deletion (InDel) markers and the rice blast resistance was studied by innoculation. The main results were as followings:
1. Genetic diversity of 150 Yunnan glutinous rice varieties with 10 individuals per variety was analyzed by 24 SSR markers that are well distributed of 12 rice chromosomes. Totally 158 alleles were detected, all SSR loci were found middle or high polymorphic. The average polymorphism information content was 0.7262. The average effective allele numbers, Shannon’s information index and expected heterozygosity were 4.5506, 1.5928 and 0.7642 respectively, which indicated abundant genetic variation of Yunnan glutinous rice. Analysis of molecular variance (AMOVA) indicated that about 86% and 14% genetic variation were detected between varieties and within varieties, respectively. Therefore, we need to protect more varieties to protect glutinous rice resources. The genetic diversity of glutinous rice varieties of south, southeast and southwest areas of Yunnan were significantly higher than that of central, northeast and northwest of Yunnan. The south, southeast and southwest Yunnan are genetic diversity centers of glutinous rice of Yunnan Province. These regions should be the hot spots of glutinous rice resource conservation. Cluster analysis showed the 150 Yunnan glutinous rice varieties contained abundant genetic variation with Nei’s genetic similarity from 0.19 to 0.99 levels, but could not be classified into groups according to geographical regions.
2. The Indica-Japonica classification and genetic variation of 60 glutinous rice varieties were done by 28 InDel markers and 24 SSR markers. The results showed that there were 24“typical Indica”, 6“Indica”, 2“Japonica”and 28“typical Japonica”varieties, respectively. By phylogeny analysis, principal component analysis and model-based structure analysis, the Indica and Japonica varieties could be separated obiously of glutinous rice. Genetic variation analysis showed the genetic diversity of Indica and Japonica glutinous rice were high, and the polymorphism difference were not significant. The analysis of molecular variance (AMOVA) indicated that 74% and 26% genetic variation were existed within and between subspecies of glutinous rice respectively with significant difference at 0.001 probability level.
3. Rice blast resistance of 100 glutinous rice varieties was studied using 8 Yunnan rice blast pathogens by manual inoculation in greenhouse. The results showed the 64 varieties were over medium susceptibility and 34 varieties were medium susceptibility. Only 2 glutinous rice varieties were found to be susceptibility. Cultivated glutinous rice varieties were relative high disease resistant.
1.选用均匀分布于水稻12条染色体的24对SSR(simple sequence repeat)引物,对来自云南13个地州36个县区的150份糯稻品种1500个单株的遗传多样性研究表明:24个SSR位点共检测到158个等位基因,每个位点平均多态信息含量为0.7262,均为中度或高度多态位点。总群体平均有效等位基因数为4.5506,平均香农指数为1.5928,平均期望杂合度为0.7642。云南糯稻总体遗传多样性水平较高。分子变异分析表明糯稻的遗传变异主要存在于品种间(86%),品种内的遗传变异占14%。保护糯稻遗传多样性需要保护较多的品种。滇南、滇东南和滇西南糯稻品种的遗传多样性显著高于滇中、滇东北和滇西北糯稻品种的遗传多样性。滇南、滇东南和滇西南是目前云南省糯稻遗传多样性中心,是保护糯稻资源时应重点关注的区域。聚类分析显示Nei遗传相似系数为0.19-0.99时150份云南糯稻品种存在丰富的遗传变异,但是不能区分地理分组。
2.用28对籼粳特异性插入缺失(insertion deletion, InDel)引物和24对简单重复序列( SSR )分析云南60份糯稻品种的籼粳分化和遗传变异,结果表明有24份糯稻品种为“典型籼型”,6份为“籼型”,2份为“粳型”,28份为“典型粳型”。聚类分析、主成分分析和基于模型的结构分析均表明糯稻籼粳分化明显,籼糯和粳糯属于两个独立的类群。遗传变异分析表明籼糯、粳糯的遗传多样性均较丰富,且SSR多态性差异不显著。分子变异分析表明糯稻的遗传变异主要来自亚种内(占总变异的74%),亚种间遗传变异占26%,但分化极显著。
3.采用温室人工接种方法用云南省的8个稻瘟病菌优势小种对100份云南糯稻品种进行抗稻瘟病鉴定,结果表明,中感以上品种和中感品种分别占到64%和34%,只有2份糯稻品种表现为感病,目前栽培糯稻品种较抗病。
The glutinous rice (Oryza sativa var. glutinosae) is the glutinosity type of rice variety that has high edible and economic value. Yunnan province is one of the richest Provinces on glutinous rice resource in China, as well as one of the origins and diversity centers of rice (Oryza sativa). In this report, the genetic diversity and structure of Yunnan glutinous rice varieties was analyzed by simple sequence repeat (SSR) markers. The Indica-Japonica differetiation was analyzed by insertion/deletion (InDel) markers and the rice blast resistance was studied by innoculation. The main results were as followings:
1. Genetic diversity of 150 Yunnan glutinous rice varieties with 10 individuals per variety was analyzed by 24 SSR markers that are well distributed of 12 rice chromosomes. Totally 158 alleles were detected, all SSR loci were found middle or high polymorphic. The average polymorphism information content was 0.7262. The average effective allele numbers, Shannon’s information index and expected heterozygosity were 4.5506, 1.5928 and 0.7642 respectively, which indicated abundant genetic variation of Yunnan glutinous rice. Analysis of molecular variance (AMOVA) indicated that about 86% and 14% genetic variation were detected between varieties and within varieties, respectively. Therefore, we need to protect more varieties to protect glutinous rice resources. The genetic diversity of glutinous rice varieties of south, southeast and southwest areas of Yunnan were significantly higher than that of central, northeast and northwest of Yunnan. The south, southeast and southwest Yunnan are genetic diversity centers of glutinous rice of Yunnan Province. These regions should be the hot spots of glutinous rice resource conservation. Cluster analysis showed the 150 Yunnan glutinous rice varieties contained abundant genetic variation with Nei’s genetic similarity from 0.19 to 0.99 levels, but could not be classified into groups according to geographical regions.
2. The Indica-Japonica classification and genetic variation of 60 glutinous rice varieties were done by 28 InDel markers and 24 SSR markers. The results showed that there were 24“typical Indica”, 6“Indica”, 2“Japonica”and 28“typical Japonica”varieties, respectively. By phylogeny analysis, principal component analysis and model-based structure analysis, the Indica and Japonica varieties could be separated obiously of glutinous rice. Genetic variation analysis showed the genetic diversity of Indica and Japonica glutinous rice were high, and the polymorphism difference were not significant. The analysis of molecular variance (AMOVA) indicated that 74% and 26% genetic variation were existed within and between subspecies of glutinous rice respectively with significant difference at 0.001 probability level.
3. Rice blast resistance of 100 glutinous rice varieties was studied using 8 Yunnan rice blast pathogens by manual inoculation in greenhouse. The results showed the 64 varieties were over medium susceptibility and 34 varieties were medium susceptibility. Only 2 glutinous rice varieties were found to be susceptibility. Cultivated glutinous rice varieties were relative high disease resistant.
引文
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