燕麦种质资源遗传多样性研究
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摘要
燕麦(Avena)属于禾本科(Gramineae)、燕麦族(Aveneae D.)、燕麦属(Avena L.)。燕麦是我国高寒山区特有的一种优势杂粮作物,是粮饲兼用兼备食疗功能的作物。研究和评价燕麦种质资源的遗传多样性,对探讨燕麦的起源和进化,种质资源的考察搜集,燕麦种质资源遗传多样性保存措施的制定,以及燕麦资源新基因的挖掘和种质创新利用均具有重要意义。本研究在内蒙古阴山南北麓2种生态条件下对74份燕麦种质资源的形态多样性、蛋白质多样性和DNA多样性等方面进行了综合评价,并对3个不同层次的遗传多样性研究进行了比较。主要结果如下:
1.对74份燕麦种质资源在2种不同生态环境下的形态多样性研究结果表明,燕麦种质资源各形态性状具有丰富的多样性,质量性状平均多样性指数为1.07;2试点数量性状平均多样性指数分别为1.90和1.65,平均变异系数为24.14%和27.66%。
2.燕麦形态性状的主成分分析和判别分析表明,供试燕麦种质的形态多样性主成分明显,呼市试点燕麦的主穗粒重、千粒重和单株粒重是造成燕麦种质形态变异的主要因素;而武川试点燕麦的主穗粒重、单株粒重和主穗小穗数是造成燕麦种质形态变异的主要因素。
3.基于形态性状聚类分析,将74份燕麦种质划分为五大种质群,供试燕麦种质类型丰富,可选到类型差异较大的不同种质材料。其中呼和浩特市试点种质群Ⅰ的材料可作为选育大粒、高产型品种的亲本,种质群Ⅱ的材料可作多穗型品种的亲本。武川试点种质群Ⅰ的材料可作为选育大粒,种质群Ⅴ的材料可作选育高产或多穗型品种的亲本,种质群Ⅲ的材料可作选育矮杆型品种的亲本。
4.利用酸性聚丙烯酰胺凝胶电泳(Acid-polyacrylamide gel electrophoresis, A-PAGE)检测与分析表明,74份燕麦醇溶蛋白位点存在丰富的等位变异,共检测到73种燕麦醇溶蛋白图谱。等位基因平均遗传多样性指数为0.5229,其醇溶蛋白带纹遗传相似系数在0.1667～1.000之间,平均遗传相似系数为0.4964。供试材料共分离出19种迁移率不同的醇溶蛋白带纹,醇溶蛋白的带纹多态性为100%。基于燕麦醇溶蛋白聚类分析将74份材料分为六类,分类结果中部分供试材料与其地理来源有一定的相关性。
5.对2个不同生态环境下74份燕麦种质资源3种营养成分进行了测定,结果表明不同种质材料之间品质性状含量存在着较为丰富的变异,呼和浩特市试点3个营养品质性状的平均变异系数和多样性指数为18.43%和1.84,武川试点3个营养品质性状的平均变异系数和多样性指数为17.32%和1.74。对其品质性状进行稳定性分析,其中编号为24、25、46、48、69、70的蛋白质含量,47、48、54的β-葡聚糖含量,13、24、25、27、71的赖氨酸含量在2试点表现含量高且稳定。并筛选出一批品质优异的燕麦资源,其中呼和浩特市32份,武川35份。
6.不同种质基因型间的蛋白质、β-葡聚糖和赖氨酸含量均达到极显著差异水平;武川试点的蛋白质和β-葡聚糖的平均含量显著高于呼和浩特市试点。裸燕麦的3个品质性状含量极显著高于皮燕麦。β-葡聚糖含量与蛋白质、赖氨酸含量呈显著正相关,蛋白质含量与赖氨酸含量均呈正相关,但不显著。
7.利用聚类分析的方法,对供试种质在2个试点的3种营养成分含量进行分类,使供试种质在不同类群间营养品质性状含量差异显著。
8.利用植物基因组试剂盒提取较高质量的燕麦DNA,其纯度高、质量好,符合燕麦AFLP分析。本研究选用核心引物3’末端添加3个选择性核苷酸的PstⅠ和MseⅠ引物,扩增出的谱带多态性比率高,效果好。
9.筛选出的10个引物组合,对74份燕麦种质的基因组进行片断多态性扩增,共扩增出784条清晰可辨的可记录带,其中760个位点具有多态性,多态性位点比率为96.91%。表明各供试材料的AFLP多态性丰富,利用AFLP标记评价燕麦种质的遗传多样性具有较高的检测效率。
10. AFLP分子标记的遗传多样性分析结果表明,燕麦种质的平均Nei'基因指数(H)为0.382,平均Shannon's信息指数为0.573,表现出较高的遗传多样性。74份燕麦品种间遗传相似系数在0.2881-0.9106之间。
11.基于AFLP分子标记的聚类分析结果,74份燕麦种质可以分为五类,分类结果与地理来源及皮裸性有相关性。
Oat (Avena sativa L.) belonging to genus Avena, tribe Aveneae, family Gramineae, was a Chinese dominated coarse crops in arid areas, and it was also an important plant of grain and forage and had food therapy function. Study and evaluation on genetic diversity of germplasm resources of oat (Avena sativa L.), which was important to the study of the origin and evolution, the investigation of germplasm resources, making protection measures for genetic diversity, new genes identification and germplasm innovation and utilization for oat. Therefore, DNA and morphological diversity and protein polymorphism of 74 oat germplasm resources were comprehensive evaluated at the northern and southern foot of Yinshan Mountain in Inner Mongolia, and compared to the study of genetic diversity of three different levels in this paper. The results were as follows.
1. Morphological diversity of 74 oats germplasm resources were studied in the two kinds of different ecological environment. The results showed that oat germplasm had rich diversity of morphological characters, with diversity index of qualitative character in average of 1.07. The average diversity index of quantitative character of two experimental sites was 1.90 and 1.65, respectively, and the average coefficient was 24.14% and 27.66%.
2. By using the principal component analysis and discrimination analysis of morphological characters of oat, the results indicated that principal components were obviously and the morphological variation of oat germplasm was caused by main spike grain weight, 1000-grain weight, grain weight per plant and seed shape at Huhhot experimental site, however, it was caused by main spike grain weight, grain weight per plant and spikelet number of main spike at Wuchuan experimental site.
3. 74 oats germplasm at two experimental sites were divided into five groups by using cluster analysis on morphological characters of oat, and the different oat germplasm with great difference in groups was selected due to very multiplicity. At Huhhot experimental site, the groupΙcan be used as parent material to select big grain and high-yield varieties, whereas the groupПcan be used as parent material to select multi spike cultivars. At Wuchuan experimental sites, as parent material, the groupΙcan be selected big varieties, the groupⅤcan be selected high-yield varieties, the groupⅤcan be selected multi spike cultivars and the groupⅢcan be selected dwarf varieties.
4. By acid-polyacrylamide gel electrophoresis (A-PAGE) analysis, the high allelic variation of gliadin alleles of 74 oats were observed, and 73 types of gliadin patterns were also detected. The average gentic diversity index of allele was 0.5229, genetic similarity coefficient of gliadin ploymorphism varied from 0.1667 to 1.000, and average genetic similarity coefficient was 0.4964. The results showed that 19 bands of gliadin were separated from materials and the gliadin ploymorphism was up to 100%. Gliadin of 74 oats were divided into six groups by cluster analysis and some of materials had related to the geographical origins including naked and covered oat.
5. Three of nutritional compositions of 74 oats germplasm resources were determined under two different ecological environments and high genetic variation in quality characteristics among different materials were observed. The average coefficient variation and diversity index of three nutritional quality characteristics was 18.43% and 1.84, respectively, at Huhhot experimental site. However, at Wuchuan experimental site, the average coefficient variation and diversity index was 17.32% and 1.74, respectively. Stability of quality characteristics were analyzed under two experimental sites, the results showed that the content of protein numbered 24, 25, 46, 48, 69 and 70,β-glucan content numbered 47, 48 and 54 and lysine content numbered 13, 24, 25, 27 and 71 was high and stable, moreover, a batch of superior quality germplasms resources were selected including 32 samples at Huhhot experimental site and 35 samples at Wuchuan experimental site.
6. The content of protein,β-glucan and lysine among different genetics of germplasm were significantly difference, and the average content of protein andβ-glucan at Wuchuan site was significantly higher than those of Huhhot site. The content of three of nutritional compositions in naked oat was significantly higher than covered oat. There was significant positive correlation between the content of protein and lysine andβ-glucan content, and there was also correlation between the content of protein and lysine, but there was not significant difference.
7. Three of nutritional compositions content of oats germplasm resources at two experimental sites were analyzed by cluster analysis and the results showed that there were significant differences in nutritional quality among different groups.
8. Extracting of high quality genome DNA from plant using plant genome kit accorded with AFLP analysis due to high purity and quality. The percentage of polymorphic bands were higher by appending Pst ? and Mse ? primer of three selective nucleotide to 3′termini of core primer in this study and had high effects.
9. 74 oats germplasm were studied by amplified fragment length polymorphism and 10 AFLP primer pairs were selected, moreover, 784 of the primers set produced clear bands and 760 of them showed polymorphism, the percentage of polymorphic bands reached to 96.91%. Results indicated that evaluating the genetic diversity of oat using AFLP marker had higher testing efficiency due to high AFLP polymorphism.
10. The results of analysis showed high genetic diversity of AFLP marker, and the average Nei’s index and Shannon's information index was 0.382 and 0.573, respectively. The variance range of genetic similarity coefficient of 74 oat varieties were 0.2881 ~ 0.9106.
11. By AFLP marker, 74 oat germplasm resources were divided into five groups by using cluster analysis, which seemed related to the geographical origins and naked or covered oat.
1.对74份燕麦种质资源在2种不同生态环境下的形态多样性研究结果表明,燕麦种质资源各形态性状具有丰富的多样性,质量性状平均多样性指数为1.07;2试点数量性状平均多样性指数分别为1.90和1.65,平均变异系数为24.14%和27.66%。
2.燕麦形态性状的主成分分析和判别分析表明,供试燕麦种质的形态多样性主成分明显,呼市试点燕麦的主穗粒重、千粒重和单株粒重是造成燕麦种质形态变异的主要因素;而武川试点燕麦的主穗粒重、单株粒重和主穗小穗数是造成燕麦种质形态变异的主要因素。
3.基于形态性状聚类分析,将74份燕麦种质划分为五大种质群,供试燕麦种质类型丰富,可选到类型差异较大的不同种质材料。其中呼和浩特市试点种质群Ⅰ的材料可作为选育大粒、高产型品种的亲本,种质群Ⅱ的材料可作多穗型品种的亲本。武川试点种质群Ⅰ的材料可作为选育大粒,种质群Ⅴ的材料可作选育高产或多穗型品种的亲本,种质群Ⅲ的材料可作选育矮杆型品种的亲本。
4.利用酸性聚丙烯酰胺凝胶电泳(Acid-polyacrylamide gel electrophoresis, A-PAGE)检测与分析表明,74份燕麦醇溶蛋白位点存在丰富的等位变异,共检测到73种燕麦醇溶蛋白图谱。等位基因平均遗传多样性指数为0.5229,其醇溶蛋白带纹遗传相似系数在0.1667～1.000之间,平均遗传相似系数为0.4964。供试材料共分离出19种迁移率不同的醇溶蛋白带纹,醇溶蛋白的带纹多态性为100%。基于燕麦醇溶蛋白聚类分析将74份材料分为六类,分类结果中部分供试材料与其地理来源有一定的相关性。
5.对2个不同生态环境下74份燕麦种质资源3种营养成分进行了测定,结果表明不同种质材料之间品质性状含量存在着较为丰富的变异,呼和浩特市试点3个营养品质性状的平均变异系数和多样性指数为18.43%和1.84,武川试点3个营养品质性状的平均变异系数和多样性指数为17.32%和1.74。对其品质性状进行稳定性分析,其中编号为24、25、46、48、69、70的蛋白质含量,47、48、54的β-葡聚糖含量,13、24、25、27、71的赖氨酸含量在2试点表现含量高且稳定。并筛选出一批品质优异的燕麦资源,其中呼和浩特市32份,武川35份。
6.不同种质基因型间的蛋白质、β-葡聚糖和赖氨酸含量均达到极显著差异水平;武川试点的蛋白质和β-葡聚糖的平均含量显著高于呼和浩特市试点。裸燕麦的3个品质性状含量极显著高于皮燕麦。β-葡聚糖含量与蛋白质、赖氨酸含量呈显著正相关,蛋白质含量与赖氨酸含量均呈正相关,但不显著。
7.利用聚类分析的方法,对供试种质在2个试点的3种营养成分含量进行分类,使供试种质在不同类群间营养品质性状含量差异显著。
8.利用植物基因组试剂盒提取较高质量的燕麦DNA,其纯度高、质量好,符合燕麦AFLP分析。本研究选用核心引物3’末端添加3个选择性核苷酸的PstⅠ和MseⅠ引物,扩增出的谱带多态性比率高,效果好。
9.筛选出的10个引物组合,对74份燕麦种质的基因组进行片断多态性扩增,共扩增出784条清晰可辨的可记录带,其中760个位点具有多态性,多态性位点比率为96.91%。表明各供试材料的AFLP多态性丰富,利用AFLP标记评价燕麦种质的遗传多样性具有较高的检测效率。
10. AFLP分子标记的遗传多样性分析结果表明,燕麦种质的平均Nei'基因指数(H)为0.382,平均Shannon's信息指数为0.573,表现出较高的遗传多样性。74份燕麦品种间遗传相似系数在0.2881-0.9106之间。
11.基于AFLP分子标记的聚类分析结果,74份燕麦种质可以分为五类,分类结果与地理来源及皮裸性有相关性。
Oat (Avena sativa L.) belonging to genus Avena, tribe Aveneae, family Gramineae, was a Chinese dominated coarse crops in arid areas, and it was also an important plant of grain and forage and had food therapy function. Study and evaluation on genetic diversity of germplasm resources of oat (Avena sativa L.), which was important to the study of the origin and evolution, the investigation of germplasm resources, making protection measures for genetic diversity, new genes identification and germplasm innovation and utilization for oat. Therefore, DNA and morphological diversity and protein polymorphism of 74 oat germplasm resources were comprehensive evaluated at the northern and southern foot of Yinshan Mountain in Inner Mongolia, and compared to the study of genetic diversity of three different levels in this paper. The results were as follows.
1. Morphological diversity of 74 oats germplasm resources were studied in the two kinds of different ecological environment. The results showed that oat germplasm had rich diversity of morphological characters, with diversity index of qualitative character in average of 1.07. The average diversity index of quantitative character of two experimental sites was 1.90 and 1.65, respectively, and the average coefficient was 24.14% and 27.66%.
2. By using the principal component analysis and discrimination analysis of morphological characters of oat, the results indicated that principal components were obviously and the morphological variation of oat germplasm was caused by main spike grain weight, 1000-grain weight, grain weight per plant and seed shape at Huhhot experimental site, however, it was caused by main spike grain weight, grain weight per plant and spikelet number of main spike at Wuchuan experimental site.
3. 74 oats germplasm at two experimental sites were divided into five groups by using cluster analysis on morphological characters of oat, and the different oat germplasm with great difference in groups was selected due to very multiplicity. At Huhhot experimental site, the groupΙcan be used as parent material to select big grain and high-yield varieties, whereas the groupПcan be used as parent material to select multi spike cultivars. At Wuchuan experimental sites, as parent material, the groupΙcan be selected big varieties, the groupⅤcan be selected high-yield varieties, the groupⅤcan be selected multi spike cultivars and the groupⅢcan be selected dwarf varieties.
4. By acid-polyacrylamide gel electrophoresis (A-PAGE) analysis, the high allelic variation of gliadin alleles of 74 oats were observed, and 73 types of gliadin patterns were also detected. The average gentic diversity index of allele was 0.5229, genetic similarity coefficient of gliadin ploymorphism varied from 0.1667 to 1.000, and average genetic similarity coefficient was 0.4964. The results showed that 19 bands of gliadin were separated from materials and the gliadin ploymorphism was up to 100%. Gliadin of 74 oats were divided into six groups by cluster analysis and some of materials had related to the geographical origins including naked and covered oat.
5. Three of nutritional compositions of 74 oats germplasm resources were determined under two different ecological environments and high genetic variation in quality characteristics among different materials were observed. The average coefficient variation and diversity index of three nutritional quality characteristics was 18.43% and 1.84, respectively, at Huhhot experimental site. However, at Wuchuan experimental site, the average coefficient variation and diversity index was 17.32% and 1.74, respectively. Stability of quality characteristics were analyzed under two experimental sites, the results showed that the content of protein numbered 24, 25, 46, 48, 69 and 70,β-glucan content numbered 47, 48 and 54 and lysine content numbered 13, 24, 25, 27 and 71 was high and stable, moreover, a batch of superior quality germplasms resources were selected including 32 samples at Huhhot experimental site and 35 samples at Wuchuan experimental site.
6. The content of protein,β-glucan and lysine among different genetics of germplasm were significantly difference, and the average content of protein andβ-glucan at Wuchuan site was significantly higher than those of Huhhot site. The content of three of nutritional compositions in naked oat was significantly higher than covered oat. There was significant positive correlation between the content of protein and lysine andβ-glucan content, and there was also correlation between the content of protein and lysine, but there was not significant difference.
7. Three of nutritional compositions content of oats germplasm resources at two experimental sites were analyzed by cluster analysis and the results showed that there were significant differences in nutritional quality among different groups.
8. Extracting of high quality genome DNA from plant using plant genome kit accorded with AFLP analysis due to high purity and quality. The percentage of polymorphic bands were higher by appending Pst ? and Mse ? primer of three selective nucleotide to 3′termini of core primer in this study and had high effects.
9. 74 oats germplasm were studied by amplified fragment length polymorphism and 10 AFLP primer pairs were selected, moreover, 784 of the primers set produced clear bands and 760 of them showed polymorphism, the percentage of polymorphic bands reached to 96.91%. Results indicated that evaluating the genetic diversity of oat using AFLP marker had higher testing efficiency due to high AFLP polymorphism.
10. The results of analysis showed high genetic diversity of AFLP marker, and the average Nei’s index and Shannon's information index was 0.382 and 0.573, respectively. The variance range of genetic similarity coefficient of 74 oat varieties were 0.2881 ~ 0.9106.
11. By AFLP marker, 74 oat germplasm resources were divided into five groups by using cluster analysis, which seemed related to the geographical origins and naked or covered oat.
引文
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