【作者】 闫伟红；

【导师】 徐柱；

【作者基本信息】 中国农业科学院 ， 草业科学， 2010， 博士

【摘要】 本论文以冰草属5种12份和鹅观草属7种27份种质材料为研究对象,采用居群取样,从形态学、生理学、醇溶蛋白、同工酶、ISSR分子标记和生产性能等六个方面,对上述种质材料进行了综合研究和分析,结果如下:1.两属供试材料在形态特征上均存在丰富的遗传变异。其中,种间变异大于种内变异。引起冰草属和鹅观草属形态变异主要有8个和9个性状。Shannon指数分析显示,光穗冰草和细茎冰草、肃草形态多样性指数最大,蒙古冰草、岷山鹅观草形态多样性指数最小;有5个种居群间形态多样性大于居群内形态多样性,有5个种与其相反;肃草地区内形态分化大于地区间形态分化,有8个种与其相反。欧氏距离聚类结果再次印证了两属植物传统分类的科学性。2.有6个生理因子影响冰草属和鹅观草属植物叶片的生理代谢功能,它们分别是丙二醛含量、电导率、可溶性糖含量、相对含水量、叶绿素b含量、可溶性蛋白质含量和丙二醛含量、电导率、脯氨酸含量、叶绿素a含量、相对含水量、可溶性蛋白质含量。在分蘖期～抽穗期,开花期～成熟期,不同物种生理代谢功能差异较大。同一物种不同材料间生理特性无显著差异。3.醇溶蛋白呈现多态性。冰草属和鹅观草属植物多态百分率分别为89.19%和92.50%。通过各遗传参数分析显示,冰草属种间遗传多样性大于种内遗传多样性,鹅观草属种间遗传多样性小于种内遗传多样性;冰草和岷山鹅观草遗传多样性最大,细茎冰草和黑药鹅观草遗传多样性最小;垂穗鹅观草地区间遗传分化小于地区内遗传分化,有5个种与其相反。4.过氧化物酶和酯酶谱带差异明显。冰草属和鹅观草属的多态性比例分别为90.00%和100.00%,其中,冰草属过氧化物酶的多态性小于酯酶,而鹅观草属正好相反。通过各遗传参数分析显示,种间遗传多样性大于种内遗传多样性;冰草和垂穗鹅观草遗传多样性最大,细茎冰草和直穗鹅观草遗传多样性最小;直穗鹅观草地区间遗传分化小于地区内遗传分化,有5个种与其相反。5.基因多样性丰富。利用ISSR分子标记技术,7条和21条随机引物从冰草属和鹅观草属供试材料中,分别检测出83条和177条多态性谱带,多态性比例分别为94.32%和92.17%。通过各遗传参数分析显示,冰草属种间遗传多样性大于种内遗传多样性,鹅观草属则相反;蒙古冰草和垂穗鹅观草遗传多样性最大,光穗冰草和黑药鹅观草遗传多样性最小;有5个种地区间遗传分化大于地区内遗传分化。6.生产性能差异明显。通过灰色关联度分析,11个和9个农艺性状与冰草属和鹅观草属产量密切相关。蒙古冰草和肃草草产量最高,冰草和直穗鹅观草种子产量最高,光穗冰草和岷山鹅观草草产量、种子产量均最低。抽穗期～开花期,不同物种生长速度有很大差异,是鉴定和评价最适宜时期。聚类结果不能完全反映两属内不同种间亲缘关系,但能揭示其生产能力的差异性。7.聚类分析显示,冰草属大部分材料基本能够聚在本种内,且聚类情况与地理来源相关。鹅观草属同种材料没有严格聚在一起,垂穗类和直穗类种质也未严格单独聚类,少部分材料聚类表现出地理同源性。从形态学、生理学、蛋白质水平和DNA分子水平获得的聚类结果基本吻合,生产性能与上述聚类结果不同。相关分析表明,不同标记水平,各多样性指数和遗传参数与原生态因子相关性不显著。8.在种间亲缘关系和系统演化上,从形态学、生理学、蛋白质水平和DNA分子水平上,推测沙生冰草很可能是冰草和蒙古冰草的天然杂交衍生种,验证光穗冰草是冰草的变种;推断小颖组垂穗鹅观草、岷山鹅观草为原始状态,长颖组肃草、多变鹅观草、直穗鹅观草为进化状态。9.根据Mantel检测结果,利用形态学、生理学、醇溶蛋白、ISSR标记和生产性能测定等研究方法,鉴定和评价冰草属种质材料,会得到比较好的效果。利用形态学、醇溶蛋白和ISSR标记等方法,鉴定和评价鹅观草属种质材料,也会收到较好的效果。同工酶的鉴定结果尚有待于进一步研究。更多还原

【Abstract】 Total of 39 germplasm materials including five species of Agropyron Gaertn. and seven species of Roegneria C.Koch. were studied in this thesis. Six methods such as morphology, physiology, gliadins, isozyme, ISSR molecular markers and production performance were used on the population samples. The aim of our study was to identify, analyze and make comprehensively research on the above germplasm materials. The results obtained were as follows:1. The materials of two genera were tested in morphology methods existed abundant genetic variation. The variation among species was greater than intraspecific variation. The largest morphological diversity was A.cristatum.var.Pectinforme, A.trachycaulu and R.stricta, the smallest morphological diversity is A.mongolicum and R.dura. There were 8 and 9 characters separately affected the morphological variation of Agropyron Gaertn. and Roegneria C.Koch.. Shannon indexes analysis showed that the morphological diversity among populations was greater than within populations in five species, but morphological diversity within populations was greater than among populations in other five species. The morphological differentiation of R.stricta in the same area was greater than it among the different areas, but the morphological differentiation of other eight species in the same area was smaller than it among the different areas. Euclidean distance clustering results confirmed the scientific traditional classification of two genera once again.2. There were six physiological factors that affected Agropyron Gaertn. and Roegneria C.Koch.leaf physiological and metabolic functions, respectively included MDA content, electrical conductivity, soluble sugar content, relative water content, chlorophyll b, soluble protein content and MDA content, electrical conductivity, proline content, chlorophyll a, relative water content, soluble protein content. From tillering period to spike period and from flowering period to mature period, physiological and metabolic functions of different species were quite different among species, physiological characteristics of different materials of the same species were no significant difference. There were have the strongest physiological ability response to environmentability to A.mongolicum and R.stricta. The related physiological factors respectively affected the yield of Agropyron Gaertn. and Roegneria C.Koch. were soluble protein content and electrical conductivity, relative water content.3. Gliadin was polymorphic. The polymorphisms percentage were 89.19% and 92.50% in Agropyron Gaertn. and Roegneria C.Koch.. The genetic diversity among species was greater than intraspecific in Agropyron Gaertn., but smaller than it in Roegneria C.Koch.. Genetic parameters analysis showed that A.cristatum and R.dura had the largest genetic diversity, A.trachycaulu and R.melanthera had the smallest genetic diversity; The gliadin bands of different populations of A.trachycaulu and R.melanthera had no significant difference; The genetic differentiation of R.nutans in the same area was greater than it among different areas, and the genetic differentiation of other five species in the same area is smaller than it among different areas.4. The bands of peroxidase and esterase had the significant difference. The polymorphism rates are 90.00% and 100.00% in Agropyron Gaertn. and Roegneria C.Koch..The polymorphism of peroxidase is greater than EST in Agropyron Gaertn., but Roegneria C.Koch. had the opposite traits. Genetic parameters analysis showed that genetic diversity among species was greater than it within species. And the maximum genetic diversity index was in A.cristatum and R.nutans, the minimum genetic diversity index is in A.trachycaulu and R.turezaninovii var. turczaninovii; The isozyme bands of different population of A.trachycaulu and R.melanthera had no significant difference; The genetic differentiation of R.turezaninovii var. turczaninovii in the same area is greater than it among different areas, and the genetic differentiation of other five species in the same area is smaller than it among different areas.5. Genetic diversity was rich. There were 83 and 177 polymorphic bands were detected respectively from 7 and 21 random primers in Agropyron Gaertn. and Roegneria C.Koch., which were tested by ISSR markers, and the polymorphism rates respectively were 94.32% and 92.17%. Genetic parameters analysis showed that Agropyron Gaertn. genetic diversity among species was greater than it within species, but Roegneria C.Koch. had the opposite traits; A.mongolicum and R.nutans have the maximum genetic diversity index, but A.cristatum. var. pectinforme and R.melanthera. had the minimum genetic diversity index. The genetic differentiation of five species in the same area is smaller than it among different areas.6. Production performance was significant difference. A.trachycaulu and R.stricta. had the best production performance. The gray correlation analysis showed that there were 11 and 9 agronomic traits closely related with the yield of Agropyron Gaertn. and Roegneria C.Koch.. The yield of grass was generally high, but seed yield was generally low. The seed yield of Agropyron Gaertn. was significantly higher than Roegneria C.Koch.. A.mongolicum and R.stricta had the highest grass yield, A.cristatum and R.turezaninovii var. turczaninovii had the highest seed yield, but A.cristatum. var. pectinforme and R.nutans. had the lowest grass yield and seed yield. Because different species had so different growth rate from spike period to flowering period, it was the optimal time to identify and evaluate the species at that time. Clustering results could not fully reflect genetic relationship of different kinds of species in two genera, but it could reveal the difference of their production performance.7. Clustering results showed that the same species of Agropyron Gaertn. could gather essentially within species and clustering conditions was related with geographical origin. The materials of the same species of Roegneria C.Koch. could not gathered within species, as the same as nutans germplasm and straight spike germplasm, but a small part of the material showed geographic clustering homology. The clustering results were consistent, which of morphology, physiology, protein and DNA molecules obtained, but production performance had different clustering results. Correlation analysis showed that the correlations among genetic diversity, parameters and ecological factors had not significant difference at different markers level.8. The genetic relationship and evolution among species analysis suggested that A.derstorum maybe a derivative species of A.cristatum and A.monogolicum, and A.cristatum. var. pectinforme was a variant of A.cristatum; It was concluded that R.nutans and R.dura which was belong to little glume group were in the original state, but R.turezaninovii var. turczaninovii which was belong to long glume group were in the evolution state.9. According to Mantel test results, it will be better to identify and evaluate the germplasm of Agropyron Gaertn. by some methods such as morphology, physiology, gliadin, ISSR markers and production performance. Use morphology, gliadin and ISSR marker methods to identify and evaluate the germplasm of Roegneria C.Koch. can also get good results. Isozyme identification still needs to be studied in the future.更多还原