苦荞资源遗传多样性研究
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摘要
苦荞是我国特有的药食同源作物,其营养价值高,富含生物类黄酮等生物活性成分,具有营养和保健的双重功效。其特有的营养组成、功能价值及地域资源特点对满足市场需求,改善人们膳食结构具有重要作用。随着人们健康意识的提高,苦荞已成为人们研究和关注的焦点。但是,苦荞属小宗作物,研究基础薄弱,当前生产利用品种存在着产量偏低、品种更换缓慢、品种应用原始单一、退化严重等一系列问题。因此,开展苦荞资源的遗传多样性研究对筛选优异种质资源,改良苦荞品种重要的指导意义。
本研究利用简单重复序列(SSR)的分子标记方法,结合形态学分析方法,对来自四川、云南、贵州、西藏、陕西、宁夏等地的205份苦荞地方品种进行遗传多样性分析,研究得出如下重要结果:
(1)农艺性状分析结果表明,205份供试材料之间差异明显。不同材料在生育期、株高、主茎分枝数、主茎节数、叶长、叶宽、主茎粗、千粒重等8个数量性状上存在差异,表现出明显的多样性。形态性状变异系数(CV%)最大的是主茎粗,达到了35.26%;其次是主茎分枝,变异系数(CV%)为23.24%,变异系数(CV%)最小的是生育期只有11.29%。通过聚类将所有苦荞地方种质资源分为四个大类,各大类之间农艺性状均值和变异系数也存在着一定的差异,但材料之间的地域性差异不明显,这可能与农艺性状受环境影响较大有关,表明每个省区的苦荞资源具有较高的遗传多样性。
(2)通过PCR扩增对30个甜荞引物和苦荞引物进行筛选,筛选出多态性比率较高的13个引物:Fes1394、Fes1816、Fes2644、Fes1286、Fes1475、Fes2743、Hyb-49、Hyb-16、Hyb-15、TH3A-GT16、TH3A-CA7、SE13-GT11、TH26-CA49,并应用于本次苦荞资源遗传多样性研究。
(3)SSR分子标记结果可以看出,所有陕西品种遗传相似系数在0.57-0.94之间,平均值为0.76;西藏的在0.65-0.93之间,平均值为0.79;云贵川地区的为0.56-0.95之间,平均值为0.76。聚类结果表明,同一地区的品种首先聚在一起。对所有供试的苦荞进行遗传多样性分析表明,205份材料的遗传相似系数变幅为0.55-0.95,平均值为0.75,品种间具有较高的遗传多样性。通过聚类将所有供试材料分为15大类,大部分材料以较多数量组成一类,其遗传距离较近,而另外一些个别材料以极少数成一类甚至单个品种成一大类,表明遗传差异较大。
(4)通过聚类结果发现,来源于陕西省宁陕的材料K047和来自云南的材料K176,独立于聚类主群体之外,单个品种成一大类。这些说明它们是较为特殊的苦荞遗传资源。进一步分析发现K047生育日数比同一地区其它品种较长,株高较低;K176生育日数比同一地区其它品种较长,千粒重较高。今后的利用当中应关注这些较特殊,且特点鲜明的品种。
Buckwheat is the important food crops and cash crops,and it also has a high nutritional value,being rich in bioflavonoids and other bioactive components and having the dual effect of nutrition and health. It is not only a honey crop,but also a traditional export commodity in china. Its unique nutritional composition,functional value and role of agricultural production play an important role to meet the current needs of people. Buckwheat has become the focus of attention,as the research of its medicinal value developments to a deeper level. The beginning study of Buckwheat is late and the progress is slow, and there is a series of serious problems exists,such as:low use of varieties,species changes slowly,the original application of a single species,serious degradation. Therefore,researches on the genetic diversity of buckwheat have an important guiding significance on screening of germplasm and improving varieties of buckwheat. In this study,by SSR molecular markers and morphological analysis, we compared the genetic polymorphisms of 205 Tartary Buckwheat Local varieties coming from Sichuan,Yunnan,Guizhou,Xizang,Shan’anxi and Ningxia. The main results are as follows:
1. The analysis of Agronomic traits indicated that: there is significant difference between all the 205 accesses. There is obvious diversity on eight Quantitative traits between all the materials. The max of Coefficient of variation of morphological traits ( CV%) is the diameter of Main stem with reaching 35.26%;Secondly is the branch of the stem with the number of 23.24%; while the growth period ranks the last reaching 11.29%. Results from Clustering showing that:all the materials can be divided into four major categories,which contains 12 sub-categories. Overall, there are obvious differences on quantitative trait between categories, but no obvious regional. Varieties of the same province had been divided into different categories showing that buckwheat from all the provinces have high genetic diversity.
2. There are 13 primers : Fes1394、Fes1816、Fes2644、Fes1286、Fes1475、Fes2743、Hyb-49、Hyb-16、Hyb-15、TH3A-GT16、TH3A-CA7、SE13-GT11、TH26-CA49 elected for the researching on genetic diversity of tartary buckwheat by PCR amplification, the 13 primers was choosed from 30 primers used on common buckwheat and tartary buckwheat.
3. The results of SSR. Analysis of genetic diversity within the same province showed that:Genetic similarity coefficient of Shananxi varieties ranged from 0.57 to 0.94, while Xizang is 0.65-0.93,and Yunnan,Guizhou,Sichuan region ranks from 0.56 to 0.95. results of clustering show that varieties of the same area always can be divided into the same categories.There is higher genetic diversity among Varieties, and the genetic similarity coefficient of all the 502 accesses ranged from 0.55 to 0.95,with an Average of 0.75. All the tested material is divided into 15 categories by clustering,most of the material form a class with a greater number showing that they have a small genetic difference,while little material even only one material form a class showing that they have a large genetic difference.
4. Results from Clustering showing that a single species can be divided into a category, such as the material K047 coming from Ningshan of Shananxi Province and K176 coming from Yunnan Province. These indicated that they are maybe the special buckwheat genetic resources. Further analysis showed that K047 have a longer fertility days and a lower height than the other varieties in the same area, while K176 have a longer fertility days and a higher grain weight than the other varieties in the same area. In the future, we should pay more attention to these species with specific and distinctive characteristics.
本研究利用简单重复序列(SSR)的分子标记方法,结合形态学分析方法,对来自四川、云南、贵州、西藏、陕西、宁夏等地的205份苦荞地方品种进行遗传多样性分析,研究得出如下重要结果:
(1)农艺性状分析结果表明,205份供试材料之间差异明显。不同材料在生育期、株高、主茎分枝数、主茎节数、叶长、叶宽、主茎粗、千粒重等8个数量性状上存在差异,表现出明显的多样性。形态性状变异系数(CV%)最大的是主茎粗,达到了35.26%;其次是主茎分枝,变异系数(CV%)为23.24%,变异系数(CV%)最小的是生育期只有11.29%。通过聚类将所有苦荞地方种质资源分为四个大类,各大类之间农艺性状均值和变异系数也存在着一定的差异,但材料之间的地域性差异不明显,这可能与农艺性状受环境影响较大有关,表明每个省区的苦荞资源具有较高的遗传多样性。
(2)通过PCR扩增对30个甜荞引物和苦荞引物进行筛选,筛选出多态性比率较高的13个引物:Fes1394、Fes1816、Fes2644、Fes1286、Fes1475、Fes2743、Hyb-49、Hyb-16、Hyb-15、TH3A-GT16、TH3A-CA7、SE13-GT11、TH26-CA49,并应用于本次苦荞资源遗传多样性研究。
(3)SSR分子标记结果可以看出,所有陕西品种遗传相似系数在0.57-0.94之间,平均值为0.76;西藏的在0.65-0.93之间,平均值为0.79;云贵川地区的为0.56-0.95之间,平均值为0.76。聚类结果表明,同一地区的品种首先聚在一起。对所有供试的苦荞进行遗传多样性分析表明,205份材料的遗传相似系数变幅为0.55-0.95,平均值为0.75,品种间具有较高的遗传多样性。通过聚类将所有供试材料分为15大类,大部分材料以较多数量组成一类,其遗传距离较近,而另外一些个别材料以极少数成一类甚至单个品种成一大类,表明遗传差异较大。
(4)通过聚类结果发现,来源于陕西省宁陕的材料K047和来自云南的材料K176,独立于聚类主群体之外,单个品种成一大类。这些说明它们是较为特殊的苦荞遗传资源。进一步分析发现K047生育日数比同一地区其它品种较长,株高较低;K176生育日数比同一地区其它品种较长,千粒重较高。今后的利用当中应关注这些较特殊,且特点鲜明的品种。
Buckwheat is the important food crops and cash crops,and it also has a high nutritional value,being rich in bioflavonoids and other bioactive components and having the dual effect of nutrition and health. It is not only a honey crop,but also a traditional export commodity in china. Its unique nutritional composition,functional value and role of agricultural production play an important role to meet the current needs of people. Buckwheat has become the focus of attention,as the research of its medicinal value developments to a deeper level. The beginning study of Buckwheat is late and the progress is slow, and there is a series of serious problems exists,such as:low use of varieties,species changes slowly,the original application of a single species,serious degradation. Therefore,researches on the genetic diversity of buckwheat have an important guiding significance on screening of germplasm and improving varieties of buckwheat. In this study,by SSR molecular markers and morphological analysis, we compared the genetic polymorphisms of 205 Tartary Buckwheat Local varieties coming from Sichuan,Yunnan,Guizhou,Xizang,Shan’anxi and Ningxia. The main results are as follows:
1. The analysis of Agronomic traits indicated that: there is significant difference between all the 205 accesses. There is obvious diversity on eight Quantitative traits between all the materials. The max of Coefficient of variation of morphological traits ( CV%) is the diameter of Main stem with reaching 35.26%;Secondly is the branch of the stem with the number of 23.24%; while the growth period ranks the last reaching 11.29%. Results from Clustering showing that:all the materials can be divided into four major categories,which contains 12 sub-categories. Overall, there are obvious differences on quantitative trait between categories, but no obvious regional. Varieties of the same province had been divided into different categories showing that buckwheat from all the provinces have high genetic diversity.
2. There are 13 primers : Fes1394、Fes1816、Fes2644、Fes1286、Fes1475、Fes2743、Hyb-49、Hyb-16、Hyb-15、TH3A-GT16、TH3A-CA7、SE13-GT11、TH26-CA49 elected for the researching on genetic diversity of tartary buckwheat by PCR amplification, the 13 primers was choosed from 30 primers used on common buckwheat and tartary buckwheat.
3. The results of SSR. Analysis of genetic diversity within the same province showed that:Genetic similarity coefficient of Shananxi varieties ranged from 0.57 to 0.94, while Xizang is 0.65-0.93,and Yunnan,Guizhou,Sichuan region ranks from 0.56 to 0.95. results of clustering show that varieties of the same area always can be divided into the same categories.There is higher genetic diversity among Varieties, and the genetic similarity coefficient of all the 502 accesses ranged from 0.55 to 0.95,with an Average of 0.75. All the tested material is divided into 15 categories by clustering,most of the material form a class with a greater number showing that they have a small genetic difference,while little material even only one material form a class showing that they have a large genetic difference.
4. Results from Clustering showing that a single species can be divided into a category, such as the material K047 coming from Ningshan of Shananxi Province and K176 coming from Yunnan Province. These indicated that they are maybe the special buckwheat genetic resources. Further analysis showed that K047 have a longer fertility days and a lower height than the other varieties in the same area, while K176 have a longer fertility days and a higher grain weight than the other varieties in the same area. In the future, we should pay more attention to these species with specific and distinctive characteristics.
引文
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