光皮树F1代早期性状遗传多样性分析与SSR-PCR体系构建
发布时间：2019-01-18 07:42
【摘要】：通过分析当前林木良种选育研究现状、应用方法及发展趋势,结合光皮树生物学生态学特性、应用价值及相关的研究基础,本研究以木本油植物光皮树(Swida wilsoniana Wanyer)7个杂交组合的亲本与子代为研究材料,采用形态学调查法、石蜡制片技术、显微观察法等方法对杂交果实种子的品质、F1代1a生、2a生、3a生幼苗的生长性状及其茎叶解剖结构特征进行多重比较及遗传多样性、相关性、遗传力分析。同时,从光皮树同科属物种SSR标记研究中筛选合适的SSR引物,构建并优化光皮树SSR-PCR分子标记体系。本研究旨在为光皮树杂交亲本的选配、目的性状的早期选择以及SSR分子标记辅助选育奠定理论及技术基础。主要研究结果如下:(1)7个杂交组合的杂交果实种子性状变幅大,变异系数在5.46%～22.17%之间。其中,千粒重变异明显大于横纵径及含油率变异;不同杂交组合种子发芽率差异较明显,变异系数为40.43%,发芽率最高的为杂交组合♂B08X早A03(41.67%),最低的是♂ 806X早B03(14.81%);杂交果实种子各性状存在不同程度的相关性,相关指数在0.608～0.930之间。发芽率与果实纵径、果实千粒重及种子纵径成极显著正相关。即饱满、纵径大的果实或者纵径较大的杂交种子发芽率相对较高。(2)F1幼苗一年生株高、地径净增长量都呈“慢～快～慢”的生长变化规律,但不同个体之间生长变化存在差异,快速生长期持续时间的长短对幼苗生长量起决定作用;二、三年生株高、地径、节间距三个性状间存在显著的正相关关系,对其中任何一性状的正向选择都将能提高其他两性状的正向选择效应;叶宽与地径也呈显著正相关,且三年生叶宽遗传力较大,遗传趋向稳定,可作为早期低强度选择指标。(3)光皮树F1幼苗茎叶解剖结构与营养生长存在一定的相关性。茎横断面积的大小主要取决于材部的面积,株高与茎横断面积呈显著正相关,与材部面积成极显著正相关;地径与叶片总厚度、海绵组织厚度呈显著正相关。因此,茎材部面积、叶海绵组织厚度可作为光皮树F1代植株生长势强弱的解剖结构预测指标。(4)建立了光皮树SSR-PCR最佳反应体系。反应总体积15μL:模板DNA用量 37.5ng、正反向引物各 0.6μmol·L-1、dNTPs 浓度为 0.4mmol·L-1、Mg2+的浓度为1.5 mmol · L-1、Taq聚合酶用量为1.5 U,引物“Cf-05”最佳退火温度为50℃。这些因素共同制约扩增的敏感性、特异性以及产量,但它们之间的影响力存在差异,从大到小分别是引物浓度Taq聚合酶模板DNA用量和dNTPs浓度Mg2+浓度;筛选获得11对适用于光皮树SSR-PCR反应体系的SSR引物,为F1分子水平遗传多样性的研究奠定了基础。
[Abstract]:Based on the analysis of the present research situation, application methods and development trends of improved tree varieties, combined with the biological and ecological characteristics, application value and related research foundation of Lithocarpus spp. In this study, the parents and progenies of seven hybrid combinations of (Swida wilsoniana Wanyer), a woody oil plant, were used as materials to study the quality of hybrid fruit seeds by means of morphological investigation, paraffin making and microscopic observation. The characters of growth and anatomical structure of 2 and 3 year old seedlings were compared and the genetic diversity, correlation and heritability were analyzed. At the same time, the suitable SSR primers were screened from the SSR markers of the same family species, and the SSR-PCR molecular marker system was constructed and optimized. The purpose of this study was to lay a theoretical and technical foundation for the selection of hybrid parents, early selection of purposive traits and SSR molecular marker-assisted selection. The main results were as follows: (1) the variation of seed characters of 7 hybrid combinations was between 5.46% and 22.17%. The variation of 1000-grain weight was higher than that of transverse diameter and oil content. The seed germination rate of different cross combinations was significantly different, the coefficient of variation was 40.43, the highest germination rate was B08X early A03 (41.67%), and the lowest was C806X early B03 (14.81%). The correlation index was between 0.6080.90 and 0.930. The germination rate was positively correlated with the vertical diameter of fruit, the weight of 1000 seeds and the longitudinal diameter of seed. That is, the fruit with full, large longitudinal diameter or hybrid seed with larger longitudinal diameter had higher germination rate. (2) the annual plant height and net increase of ground diameter of F1 seedlings were all "slow-fast ~ slow-slow". However, there were differences in growth changes among different individuals, and the duration of rapid growth period played a decisive role in seedling growth. (2) there were significant positive correlations among the three traits of plant height, ground diameter and pitch, and the positive selection of any one of the traits would increase the positive selection effect of other hermaphrodite traits. The leaf width was also positively correlated with the basal diameter, and the heritability of leaf width was higher and the genetic tendency was stable. (3) there was a certain correlation between the stem and leaf anatomical structure of F1 seedlings and the vegetative growth. The size of stem cross-sectional area mainly depends on the area of wood, plant height is significantly positive correlation with stem cross-sectional area, and extremely significant positive correlation with wood area, the root diameter and leaf total thickness, sponge tissue thickness has a significant positive correlation. Therefore, the area of stem material and the thickness of leaf sponge can be used as anatomic structure indexes to predict the growth potential of F1 plants. (4) the best reaction system of SSR-PCR was established. The total volume of the reaction was 15 渭 L: the dosage of template DNA was 37.5 ng / g, and the concentration of 0.4mmol L-1nNTPs was 1.5 mmol L -1 Taq polymerase. The optimum annealing temperature of primer "Cf-05" is 50 鈩，
本文编号：2410473
[Abstract]:Based on the analysis of the present research situation, application methods and development trends of improved tree varieties, combined with the biological and ecological characteristics, application value and related research foundation of Lithocarpus spp. In this study, the parents and progenies of seven hybrid combinations of (Swida wilsoniana Wanyer), a woody oil plant, were used as materials to study the quality of hybrid fruit seeds by means of morphological investigation, paraffin making and microscopic observation. The characters of growth and anatomical structure of 2 and 3 year old seedlings were compared and the genetic diversity, correlation and heritability were analyzed. At the same time, the suitable SSR primers were screened from the SSR markers of the same family species, and the SSR-PCR molecular marker system was constructed and optimized. The purpose of this study was to lay a theoretical and technical foundation for the selection of hybrid parents, early selection of purposive traits and SSR molecular marker-assisted selection. The main results were as follows: (1) the variation of seed characters of 7 hybrid combinations was between 5.46% and 22.17%. The variation of 1000-grain weight was higher than that of transverse diameter and oil content. The seed germination rate of different cross combinations was significantly different, the coefficient of variation was 40.43, the highest germination rate was B08X early A03 (41.67%), and the lowest was C806X early B03 (14.81%). The correlation index was between 0.6080.90 and 0.930. The germination rate was positively correlated with the vertical diameter of fruit, the weight of 1000 seeds and the longitudinal diameter of seed. That is, the fruit with full, large longitudinal diameter or hybrid seed with larger longitudinal diameter had higher germination rate. (2) the annual plant height and net increase of ground diameter of F1 seedlings were all "slow-fast ~ slow-slow". However, there were differences in growth changes among different individuals, and the duration of rapid growth period played a decisive role in seedling growth. (2) there were significant positive correlations among the three traits of plant height, ground diameter and pitch, and the positive selection of any one of the traits would increase the positive selection effect of other hermaphrodite traits. The leaf width was also positively correlated with the basal diameter, and the heritability of leaf width was higher and the genetic tendency was stable. (3) there was a certain correlation between the stem and leaf anatomical structure of F1 seedlings and the vegetative growth. The size of stem cross-sectional area mainly depends on the area of wood, plant height is significantly positive correlation with stem cross-sectional area, and extremely significant positive correlation with wood area, the root diameter and leaf total thickness, sponge tissue thickness has a significant positive correlation. Therefore, the area of stem material and the thickness of leaf sponge can be used as anatomic structure indexes to predict the growth potential of F1 plants. (4) the best reaction system of SSR-PCR was established. The total volume of the reaction was 15 渭 L: the dosage of template DNA was 37.5 ng / g, and the concentration of 0.4mmol L-1nNTPs was 1.5 mmol L -1 Taq polymerase. The optimum annealing temperature of primer "Cf-05" is 50 鈩，
本文编号：2410473
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