【作者】 时剑；

【导师】 童再康；

【作者基本信息】 浙江农林大学 ， 林木遗传育种， 2011， 硕士

【摘要】 石蒜属植物是一类优良的观赏植物,因其观赏性状变异丰富,具有巨大的开发潜力。本研究利用常规育种和分子标记辅助选育等手段,开展观赏新品种选育技术研究,建立了中国石蒜的SSR标记体系,对其特殊观赏性状进行试验研究;选取石蒜和换锦花二个代表性观赏种,开展鳞茎的组培快繁技术和高效培育技术研究。结果如下:1.筛选出一批具有特殊观赏价值的种质材料（无性系）,包括:中国石蒜61个,石蒜37个,换锦花24个以及少量的长筒石蒜、稻草石蒜、江苏石蒜、忽地笑、乳白石蒜等,共计138个,并繁殖成无性系,对这些无性系的形态、物候进行观测,获得第一手资料,为进一步的新品种选育和品种审定工作提供支持。2.对石蒜等5个物种,开展种间杂交试验研究,并获得一定数量的杂交种苗,结果表明:可育的二倍体亲本群体杂交结实率在0.86².92之间,正反交试验的分析表明,母本在杂交结实率的大小上起着决定性的作用。3.建立了中国石蒜的SSR分子标记体系:20μL含Taq酶0.5U、Mg²⁺ 1.5mmol·L^-1、dNTP 0.125mmol·L^-1、Primer 0.5mmol·L^-1、DNA 50 ng。利用该体系对石蒜属的8个物种和中国石蒜种内35个无性系进行扩增,每对引物在种间平均得到7个多态性位点,在中国石蒜种内得到9.5个多态性位点。对中国石蒜的花葶高度、开花期等性状和SSR扩增条带之间的对应关系进行分析,得到与迟花期性状相关的Ly97、Ly98条带,与中矮花葶高度性状相关的Ly84、Ly87、Ly97条带等。4.以换锦花种胚和叶片为外植体进行组织培养研究,结果显示:换锦花种胚的诱导率较高,平均为75.8%,适宜的愈伤组织诱导培养基为MS+6-BA 2mg·L^-1+2,4-D 2 mg·L^-1+蔗糖60g·L^-1,适宜的丛芽诱导培养基为MS+6-BA 2mg·L^-1+2,4-D 1.0 mg·L^-1+蔗糖30g·L^-1;换锦花幼嫩叶片的平均诱导率为65.8%,适宜于幼嫩叶片诱导的培养基为MS+6-BA 20mg·L^-1+NAA 1mg·L^-1+蔗糖30g·L^-1。5.对石蒜栽培试验中土壤营养、叶片营养与生长的关系等进行研究,结果显示:石蒜鳞茎球重增量和围径增量随施肥量的增加呈现明显的单峰曲线变化趋势,鳞茎的最大生长量比对照组增产15.7%;确定了依据土壤肥力状况的施肥量模型;利用叶分析法对石蒜营养诊断方法进行尝试,得到叶片氮磷钾营养元素的临界浓度为3.05%、19.40%、31.29%,维持一个高于该浓度的营养水平对保证产量至关重要。更多还原

【Abstract】 Lycoris Herb. is one of the most important flowering bulbs, widely distributed in the southern subtropical regions of China,and has great potential for the development and utilization because of its unusual flower and numerous variation. Conventional breeding and molecular marker-assisted breeding methods were carried on the breeding of new varieties in this article. SSR molecular marker analysis protocol was established and was used on the correspondence analysis of fancypoints in L. chinensis.Tissue culture of L.sprengeri and the bulb cultivation techniques of L.radiata were studied respectively. Results are as follows:1. A number of clones with special ornamental value were selected as follows: 61 L. chinensis, 37 L. radiata, 24 L.sprengeri and a small amount of L.longituba, L. straminea, L. houdyshelii, L. aurea, L. albiflora, and a total of 138. First-hand information of these clones was obatianed through observations which provide support for cultivar assessment.2. Inter-species hybridization among 5 species was carried out. Results showed that: hybridization among fertile diploid hybrid parent groups was easy. And its seed between 0.86².92 on average, which was below compare with fertile diploid hybrid parent groups except L. chinensis. Analysis of reciprocal cross experiments showed that the number of hybrid seed depends on its female parent.3. An SSR molecular marker analysis protocol was established in L. chinensis, which had a total volume of 20μL containing 0.5 U of Taq DNA polymerase, 1.5 mmol·L^-1 Mg²⁺, 0.125 mmol·L^-1 dNTP, 0.5mmol·L^-1 primer and 50 ng of template DNA. This protocol had been applied to an analysis of 8 species of Lycoris and 35 clones of L. chinensis with 7 and 9.5 polymorphic loci per primer on average obtained, respectively, which reflects a good performance in universality and stability of this protocol. Analysis of the corresponding relation between SSR loci and such traits as scape height and flowering time had preliminarily shown that loci Ly 97 and Ly 98 were correlated with late flowering time‘Oct’as well as that loci Ly 84, Ly 87 and Ly 97 with scape heights‘Mid’and‘Low’.4. The leaf blades and embryo of L.sprengeri were cultured in vitro.The results showed that high induction frequency, 75.8% on average, was obtained on embryo culture. The most ideal callus induction medium of embryo culture was MS+6-BA 2mg·L^-1+2,4-D 2 mg·L^-1+ Sucrose 60g·L^-1,while the most ideal clustered shoots induction medium was MS+6-BA 2mg·L^-1+2,4-D 1 mg·L^-1+ Sucrose 30g·L^-1. The results of leaf tissue culture showed that the induction frequency of young leaves was 65.8% on average which was higher than that of mature leaves（13.5% on average）. The most ideal induction medium of young leaves tissue culture was MS+6-BA 20mg·L^-1+NAA 1mg·L^-1+Sucrose 30g·L^-1。5. The effects of fertilizing amount on L. radiata were studied, including the soil nutrient, blade nutrient, bulb weigh and circumference. The result indicated that the bulb weight and circumference increment trends of change in different fertilizing amount treatments present unimodal curve. The maximum bulb growth was higher than control group with 15.7%, while, excessive fertilizing amount stunted the growth. A fertilizing model was evaluated on the basis of soil fertility. Leaf analysis was studied on nutrition diagnosis method. The result showed that the critical concentration of leaf nutrient element was 3.05%、19.40%、31.29% respectively on N、P₂O₅、K₂O,above which was necessary on ensuring a high output.更多还原