木薯种质资源的遗传多样性分析与评价
摘要
木薯是热带地区多年生块根作物,是世界三大薯类作物之一。也是热带、亚热带地区六亿人赖以生存的主要食物来源,淀粉含量高,其块根淀粉含量在27-34%之间,被誉为“淀粉之王”。目前,国家制定的再生能源发展战略和新颁布的《国家可再生能源法》中,将木薯列为未来生产燃料酒精的首选原料之一。木薯已成为全球重要的粮食和新型能源作物,木薯种质资源的分类与评价是育种和生产利用的基础。
本研究采用SRAP分子标记对134份木薯种质的遗传多样性进行分析,使用NTSYS2.1软件从分子水平上揭示了供试材料的遗传分类关系,分组群体间的遗传多样性水平及其亲缘关系,并对18份主要的栽培品种和国外引进高淀粉材料进行了DNA指纹图谱的绘制。本研究的主要结论如下：
(1)36对SRAP引物对134份种质群体扩增共获得306个多态性标记,多态性百分率为87.9%,平均多态性带数为8.5个,适合群体的遗传多样性分析。
(2)聚类分析将全部材料划分为8个组群,将为木薯杂交育种亲本选择提供基本的遗传差异信息；结合PCA分析说明,分组群体构成与其品系的地理来源具有一般相关性,但是存在一些交叉,初步说明我国品种引进后发生部分地理演化。
(3)遗传多样性参数统计表明,组间遗传多样性水平有了明显的上升,达到0.460,但是各个组内存在差异,以第1和第5组较高,它们分别是由我国的自育品种和来自南美洲的品系构成。说明引种后我国木薯种质库的整体遗传多样性水平较为丰富,为杂交育种亲本选择提供了依据,但是组内遗传多样性仍然需要进一步扩展。组间遗传分化和基因流水平较低。
(4)初步分析了分子标记聚类与表型聚类之间的对应关系,发现两种聚类结果存在较大的差异。
(5)采用2对SRAP引物组合Em24-Me10和Em1-Me5,绘制了18个重要木薯品种的DNA指纹图谱,其置信概率达到99.99%,对品种知识产权保护和鉴别具有重要意义。
Cassava is a perennial root crop in the tropics, one of the three major tuberous crops in the world. And the main foodstuff of 600 million people in tropical and subtropical regions, and the root has high starch content (about 27-34%), named as "king of starch". At present, China's renewable energy development strategy, and the newly published " National Renewable Energy Law" list cassava as one of the raw materials for bio-alcohol. Now, cassava has become an important food and new energy crops in the world, cassava germplasm classification and evaluating is the basis job for breeding and agricultural production.
In this study, we applied Sequence-Related Amplified Polymorphism (SRAP) technology to analyze the genetic diversity of 134 cassava varieties/lines, and revealed the genetic cluster relationship among them by NTSYS2.1. Then we calculated the genetic diversity and the affinity in different groups on molecular level. Furthermore, we draw a DNA Fingerprinting draft for 18 varieties/lines which including the dominants cultivars and high-starch foreign cassava lines. The main results of this study as follows:
(1) 36 SRAP pairs primers were selected to screen 134 cassava varieties/lines, totally 306 polymorphic bands were generated, and its ratio is 87.9%, average 8.5 polymorphic band per primer, it is suitable to analyze genetic diversity in cassava group.
(2) All material were divided into 8 group by genetic cluster analyzing,will afford basic message of genetic diversity for cassava cross-breeding. Combining with the results of PCA analysis, it showed that there was general relevance between group composition and geographical origin among these 134 varieties/lines. But part of varieties/lines doesn't obey the rules, and the preliminary results in this study showed that China cassava germpalsm has partly geographical evolution after being introduced.
(3) The genetic diversity between populations had significant increased by estimate the genetic parameter among different groups, reached to 0.46. But the genetic parameter of different groups was not consistency, group1 and group 5 that consisted with Chinese varieties and South American lines had higher parameter than others. That means after introduction the cassava germplasm resource in China of genetic diversity is more abundant, for choosing of hybrid breeding parents provide a basis, but genetic diversity in groups should be expanding. Gene differentiation and gene flow between groups in lower level.
(4) Preliminary analysis of the relationship between molecular markers and phenotypic clustering, results showed that the inconsistence was existed.
(5) DNA fingerprint draft of 18 important cassava varieties was by using primer combination Mel-Em5 and Me24-Em10, the confidence level was 99.99%. And it is an important for intellectual property protection and identification of the cassava species.
本研究采用SRAP分子标记对134份木薯种质的遗传多样性进行分析,使用NTSYS2.1软件从分子水平上揭示了供试材料的遗传分类关系,分组群体间的遗传多样性水平及其亲缘关系,并对18份主要的栽培品种和国外引进高淀粉材料进行了DNA指纹图谱的绘制。本研究的主要结论如下：
(1)36对SRAP引物对134份种质群体扩增共获得306个多态性标记,多态性百分率为87.9%,平均多态性带数为8.5个,适合群体的遗传多样性分析。
(2)聚类分析将全部材料划分为8个组群,将为木薯杂交育种亲本选择提供基本的遗传差异信息；结合PCA分析说明,分组群体构成与其品系的地理来源具有一般相关性,但是存在一些交叉,初步说明我国品种引进后发生部分地理演化。
(3)遗传多样性参数统计表明,组间遗传多样性水平有了明显的上升,达到0.460,但是各个组内存在差异,以第1和第5组较高,它们分别是由我国的自育品种和来自南美洲的品系构成。说明引种后我国木薯种质库的整体遗传多样性水平较为丰富,为杂交育种亲本选择提供了依据,但是组内遗传多样性仍然需要进一步扩展。组间遗传分化和基因流水平较低。
(4)初步分析了分子标记聚类与表型聚类之间的对应关系,发现两种聚类结果存在较大的差异。
(5)采用2对SRAP引物组合Em24-Me10和Em1-Me5,绘制了18个重要木薯品种的DNA指纹图谱,其置信概率达到99.99%,对品种知识产权保护和鉴别具有重要意义。
Cassava is a perennial root crop in the tropics, one of the three major tuberous crops in the world. And the main foodstuff of 600 million people in tropical and subtropical regions, and the root has high starch content (about 27-34%), named as "king of starch". At present, China's renewable energy development strategy, and the newly published " National Renewable Energy Law" list cassava as one of the raw materials for bio-alcohol. Now, cassava has become an important food and new energy crops in the world, cassava germplasm classification and evaluating is the basis job for breeding and agricultural production.
In this study, we applied Sequence-Related Amplified Polymorphism (SRAP) technology to analyze the genetic diversity of 134 cassava varieties/lines, and revealed the genetic cluster relationship among them by NTSYS2.1. Then we calculated the genetic diversity and the affinity in different groups on molecular level. Furthermore, we draw a DNA Fingerprinting draft for 18 varieties/lines which including the dominants cultivars and high-starch foreign cassava lines. The main results of this study as follows:
(1) 36 SRAP pairs primers were selected to screen 134 cassava varieties/lines, totally 306 polymorphic bands were generated, and its ratio is 87.9%, average 8.5 polymorphic band per primer, it is suitable to analyze genetic diversity in cassava group.
(2) All material were divided into 8 group by genetic cluster analyzing,will afford basic message of genetic diversity for cassava cross-breeding. Combining with the results of PCA analysis, it showed that there was general relevance between group composition and geographical origin among these 134 varieties/lines. But part of varieties/lines doesn't obey the rules, and the preliminary results in this study showed that China cassava germpalsm has partly geographical evolution after being introduced.
(3) The genetic diversity between populations had significant increased by estimate the genetic parameter among different groups, reached to 0.46. But the genetic parameter of different groups was not consistency, group1 and group 5 that consisted with Chinese varieties and South American lines had higher parameter than others. That means after introduction the cassava germplasm resource in China of genetic diversity is more abundant, for choosing of hybrid breeding parents provide a basis, but genetic diversity in groups should be expanding. Gene differentiation and gene flow between groups in lower level.
(4) Preliminary analysis of the relationship between molecular markers and phenotypic clustering, results showed that the inconsistence was existed.
(5) DNA fingerprint draft of 18 important cassava varieties was by using primer combination Mel-Em5 and Me24-Em10, the confidence level was 99.99%. And it is an important for intellectual property protection and identification of the cassava species.
引文
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