【摘要】：通過分析當前林木良種選育研究現(xiàn)狀、應用方法及發(fā)展趨勢,結(jié)合光皮樹生物學生態(tài)學特性、應用價值及相關的研究基礎,本研究以木本油植物光皮樹(Swida wilsoniana Wanyer)7個雜交組合的親本與子代為研究材料,采用形態(tài)學調(diào)查法、石蠟制片技術(shù)、顯微觀察法等方法對雜交果實種子的品質(zhì)、F1代1a生、2a生、3a生幼苗的生長性狀及其莖葉解剖結(jié)構(gòu)特征進行多重比較及遺傳多樣性、相關性、遺傳力分析。同時,從光皮樹同科屬物種SSR標記研究中篩選合適的SSR引物,構(gòu)建并優(yōu)化光皮樹SSR-PCR分子標記體系。本研究旨在為光皮樹雜交親本的選配、目的性狀的早期選擇以及SSR分子標記輔助選育奠定理論及技術(shù)基礎。主要研究結(jié)果如下:(1)7個雜交組合的雜交果實種子性狀變幅大,變異系數(shù)在5.46%～22.17%之間。其中,千粒重變異明顯大于橫縱徑及含油率變異;不同雜交組合種子發(fā)芽率差異較明顯,變異系數(shù)為40.43%,發(fā)芽率最高的為雜交組合♂B08X早A03(41.67%),最低的是♂ 806X早B03(14.81%);雜交果實種子各性狀存在不同程度的相關性,相關指數(shù)在0.608～0.930之間。發(fā)芽率與果實縱徑、果實千粒重及種子縱徑成極顯著正相關。即飽滿、縱徑大的果實或者縱徑較大的雜交種子發(fā)芽率相對較高。(2)F1幼苗一年生株高、地徑凈增長量都呈“慢～快～慢”的生長變化規(guī)律,但不同個體之間生長變化存在差異,快速生長期持續(xù)時間的長短對幼苗生長量起決定作用;二、三年生株高、地徑、節(jié)間距三個性狀間存在顯著的正相關關系,對其中任何一性狀的正向選擇都將能提高其他兩性狀的正向選擇效應;葉寬與地徑也呈顯著正相關,且三年生葉寬遺傳力較大,遺傳趨向穩(wěn)定,可作為早期低強度選擇指標。(3)光皮樹F1幼苗莖葉解剖結(jié)構(gòu)與營養(yǎng)生長存在一定的相關性。莖橫斷面積的大小主要取決于材部的面積,株高與莖橫斷面積呈顯著正相關,與材部面積成極顯著正相關;地徑與葉片總厚度、海綿組織厚度呈顯著正相關。因此,莖材部面積、葉海綿組織厚度可作為光皮樹F1代植株生長勢強弱的解剖結(jié)構(gòu)預測指標。(4)建立了光皮樹SSR-PCR最佳反應體系。反應總體積15μL:模板DNA用量 37.5ng、正反向引物各 0.6μmol·L-1、dNTPs 濃度為 0.4mmol·L-1、Mg2+的濃度為1.5 mmol · L-1、Taq聚合酶用量為1.5 U,引物“Cf-05”最佳退火溫度為50℃。這些因素共同制約擴增的敏感性、特異性以及產(chǎn)量,但它們之間的影響力存在差異,從大到小分別是引物濃度Taq聚合酶模板DNA用量和dNTPs濃度Mg2+濃度;篩選獲得11對適用于光皮樹SSR-PCR反應體系的SSR引物,為F1分子水平遺傳多樣性的研究奠定了基礎。
[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，

本文編號：2410473