基于两种分子标记鉴定甜菜品种及杂交组合一致性的研究

【作者】 王莹；

【导师】 吴则东；

【作者基本信息】 黑龙江大学 ， 植物保护， 2025， 硕士

【摘要】 为实现分子标记技术对甜菜品种及杂交组合一致性的科学精准快速鉴定,明晰甜菜品种在分子层面的一致性水平,本研究致力于初步构建甜菜品种一致性鉴定标准,从而为甜菜新品种登记与保护工作提供理论支撑。利用6份甜菜品种对SSR引物和InDel引物进行了筛选,基于筛选出来的核心引物对21个已登记甜菜品种及20个杂交组合开展一致性鉴定工作,初步给出了一套用于分子标记鉴定甜菜品种一致性的标准,以期为甜菜品种的一致性快速鉴定提供新方法,为甜菜新品种的登记和保护提供参考依据。研究结果如下:1.利用6个来甜菜品种的基因组DNA模板对58对SSR引物和70对InDel引物进行筛选。筛选出适用于甜菜分子标记的多态性高且重复性好的18对SSR引物和18对InDel引物,这些引物的PIC值均大于0.5,且每种引物在每条染色体上均为两条。2.通过18对SSR引物对21份甜菜登记品种进行一致性分析:12个三交种平均一致性比率在0.65～0.88之间,其中KWS1197的一致性最好,其R值最大,为0.88。9个单交种平均一致性比率在0.64～0.78之间,其中NT39106的一致性最好,其R值最大,为0.78。通过18对SSR引物对20份甜菜杂交组合进行一致性分析:其中10个单交种杂交组合平均一致性比率在0.72～0.83之间,其中XJT2219的一致性最好,其R值最大,为0.83。10个三交种杂交组合平均一致性比率在0.62～0.86之间,其中SR23014的一致性最好,其R值最大,为0.86。3.通过18对InDel引物对21份甜菜登记品种进行一致性分析:其中12个三交种平均一致性比率在0.70～0.87之间,其中HI0474的一致性最好,其R值最大,为0.87。9个单交种平均一致性比率在0.72～0.93之间,其中XJT9917的一致性最好,其R值最大,为0.93。STS21905一致性相较其他品种中等,其R值为0.72。通过18对InDel引物对20份甜菜杂交组合进行一致性分析,其中10个单交种杂交组合平均一致性比率在0.64～0.85之间,其中XJT2163的一致性最好,其R值最大,为0.85。XJT2209在这10个杂交组合中一致性相对较差,其R值为0.64。10个三交种杂交组合平均一致性比率在0.72～0.81之间,其中HD206和SR23013的一致性最好,其R值最大,为0.81。4.分析鉴定甜菜品种的一致性,需要参考所有引物位点的平均一致性比率R和单个引物的一致性比率r两个指标分析评价。综合两种分子标记对41份甜菜材料的一致性鉴定结果,参考其他作物品种分子标记的一致性鉴定的文献,再结合甜菜的自交不亲和性以及甜菜三交种的特点,初步确定甜菜品种鉴定一致性的评价方法,将其划分为5级,为将来甜菜品种一致性的鉴定提供参考。更多还原

【Abstract】 In order to realize the scientific,precise and rapid identification of the consistency of sugar beet varieties and hybrid combinations by molecular marker technology,and to clarify the consistency level of sugar beet varieties at the molecular level,the present study is dedicated to the preliminary construction of the consistency identification standard of sugar beet varieties,so as to provide theoretical support for the registration and protection of new varieties of sugar beet.Six sugar beet varieties were screened for SSR primers and InDel primers,and 21 registered sugar beet varieties and 20 hybrid combinations were identified for consistency based on the screened core primers,and a set of standards for identifying the consistency of sugar beet varieties with molecular markers was initially given,with the aim of providing a new method for the rapid identification of consistency of sugar beet varieties,and providing a reference basis for the registration and protection of new varieties of sugar beet.The results of the study are as follows.The results of the study are as follows:1.58 pairs of SSR primers and 70 pairs of InDel primers were screened using genomic DNA templates from six sugar beet varieties.Eighteen pairs of SSR primers and18 pairs of InDel primers with high polymorphism and good reproducibility suitable for molecular labeling in sugar beet were screened,and the PIC values of these primers were all greater than 0.5,and each primer was two on each chromosome.2.Consistency analysis of 21 registered sugar beet varieties was carried out by 18pairs of SSR primers:the average consistency ratio of 12 tri-crosses ranged from 0.65 to0.88,with KWS1197 having the best consistency and the largest R-value of 0.88.The average consistency ratio of 9 single-crosses ranged from 0.64 to 0.78,with NT39106having the best consistency and the largest R-value of 0.78.The consistency analysis of20 sugar beet hybrid combinations was carried out by 18 pairs of SSR primers:the average consistency ratios of 10 single-cross hybrid combinations ranged from 0.72 to0.83,among which the consistency of XJT2219 was the best,with the largest R-value of0.83.The average consistency ratios of 10 three-cross hybrid combinations ranged from0.62 to 0.86,among which the consistency of SR23014 was the best,with the largest R-value of 0.83.SR23014 had the best consistency with the largest R-value of 0.86.3.Consistency analysis of 21 registered sugar beet varieties by 18 pairs of InDel primers:12 tri-crosses had average consistency ratios ranging from 0.70 to 0.87,with HI0474 having the best consistency with the largest R-value of 0.87.9 single-crosses had average consistency ratios ranging from 0.72 to 0.93,with XJT9917 having the best consistency with the largest R-value of 0.93.STS21905 had moderate consistency compared to the other varieties,with an R-value of 0.72.The consistency of STS21905was moderate compared with other varieties,with an R value of 0.72.Consistency analysis of 20 sugar beet hybrid combinations was carried out by 18 pairs of InDel primers,and the average consistency ratios of the 10 single-cross hybrid combinations ranged from0.64 to 0.85,among which the consistency of XJT2163 was the best,with the largest R value of 0.85.The consistency of XJT2209 in these combinations ranged from 0.64 to0.85,and that of XJT2209 was the best,with an R value of 0.85.XJT2209 had relatively poor consistency among these 10 hybrid combinations,with an R value of 0.64.The average consistency ratios of the 10 three-cross hybrid combinations ranged from 0.72 to0.81,with HD206 and SR23013 having the best consistency,with the largest R value of0.81.4.To analyze and identify the consistency of sugar beet varieties,it is necessary to refer to the average consistency ratio R of all primer sites and the consistency ratio r of individual primers.By synthesizing the results of consistency identification of two molecular markers on 41 sugar beet materials,referring to the literature on consistency identification of molecular markers of other crop varieties,and combining the self-incompatibility of sugar beet and the characteristics of sugar beet tri-crosses,we initially determined the evaluation method of consistency of sugar beet varieties identification,and classified it into five levels,which will provide a reference for the identification of consistency of sugar beet varieties in the future.更多还原