广东野百合DNA提取和RAPD条件的优化

以野百合(Lilium brownii)新鲜叶片、硅胶干燥叶片及鳞片为材料,研究了DNA的提取方法,并对影响随机扩增多态DNA(RAPD)反应的各因素进行了优化。建立了野百合RAPD的优化反应体系及程序,即在20μl反应体系中,含20 ng模板DNA,2.0 mmol/L Mg2 、0.2 mmol/L dNTPs、1.5 U Taq DNA聚合酶、0.3μmol/L随机引物S1519;扩增程序为:94℃预变性5 min,然后94℃30 s,38℃50 s,72℃1 min,35个循环,最后72℃延伸10 min,4℃保存。     

山丹不同居群花器官的形态多样性研究

为掌握山丹种质资源花器官的多样性,对国内山丹资源进行引种种植观测。结果表明,25个山丹居群的花器官性状表现出一定差异。差异主要来源于花蕾毛的有无、花瓣宽、反卷度及花色等性状,其中花朵直径、花药长与地理因子(经度、纬度、海拔)存在显著或极显著相关性;根据主要花器官性状对25个居群进行类平均UPGMA聚类,可以分为5类,结果表明山丹居群花器官性状的聚类与地理位置及海拔都有一定的关系。     

大花卷丹的组织培养及限制生长保存

离体保存是植物种质保存的重要手段之一,为实现对大花卷丹的保护性利用,本文对其组织培养体系及限制生长离体保存技术进行了研究。结果表明,在常温（23±2）℃、光照强度约为40μmol.m-2.s-1、光照时间14h.d-1的条件下,大花卷丹鳞片在MS＋6-BA1.0mg.L-1＋NAA0.2mg.L-1培养基中生长情况较好,能直接诱导芽,且小鳞茎的生长速度较快。将诱导出的小鳞茎切割后,接种到1/2MS＋NAA0.5mg.L-1＋活性炭2g.L-1生根培养基2～3周即能生根,生长状况良好。提高MS培养基中蔗糖达90和110g.L-1时可以抑制其生长,能够保存大花卷丹试管苗10个月,保存过程中生长正常,株高生长缓慢,但根长势较快。在蔗糖浓度90g.L-1基础上再添加30g.L-1甘露醇的培养基能进一步抑制试管苗根的生长。6个月后,转移到正常培养基上培养均能恢复生长,其鳞片在诱导培养基上能正常分化。因此,采用大花卷丹鳞片组织培养可以形成种苗,在培养基中添加高蔗糖浓度和甘露醇可以使其试管苗保存1年以上。     

暗期光间断条件下新铁炮百合花芽分化的形态学研究

在暗期光间断条件下采用石蜡切片技术对新铁炮百合花芽分化过程进行了研究.结果表明:新铁炮百合品种花芽或顶芽分化过程可分为营养生长期、花原基或花序原基分化期、花被分化期、雌雄蕊分化期和花序形成期,而对于新铁炮百合品种花器官的发生过程而言只具有前4个时期,而无花序形成期;暗期光间断处理下80％新铁炮百合花序形成所跨度的时间为25 d.研究结果可为新铁炮百合在生产上进行促花管理控制夜间光照天数提供有益参考.     

不同光周期条件下新铁炮百合生理指标的对比研究

在长、短两种光周期条件下对新铁炮百合的三种生理指标进行了测定。结果表明:新铁炮百合上、中部叶片的生理活动与植株生长发育状态的关系较为紧密;顶芽形态分化初期,上、中部叶片可溶性糖和淀粉含量降至较低的水平,而总氮含量则升至较高水平;此外,生殖生长状态下,3部分叶片各自C/N值均高于营养生长状态下其各自C/N值。     

淡黄花百合珠芽发育过程的形态学与解剖学研究

以野生淡黄花百合(Lilium sulphureum)为实验材料,通过形态学观察、石蜡切片方法对珠芽的发生规律、发育过程中的形态学变化及解剖结构特征进行了研究,以阐明淡黄花百合珠芽繁殖的生物学机制。结果显示:淡黄花百合的珠芽形成于地上茎叶腋处叶柄的近基部位置,由叶柄表皮以内数层薄壁细胞不断分裂和分化形成;珠芽的发育过程可划分为启动期、膨大期和成熟期三个时期。成熟珠芽外观上成圆形或椭圆形,珠芽中部的薄壁细胞含有大量淀粉颗粒。淡黄花百合的珠芽具有很高的萌发率(90%以上)。研究表明,珠芽营养繁殖是其生态适应的一种重要机制。     

百合不定芽培养脱毒种球生产的研究

百合不定芽培养再生植株的病毒脱毒效果因百合品种和病毒种类而有所差异;‘卡萨布兰卡’百合潜在病毒(LSV)容易脱去,脱毒率达76％,‘魅丽’黄瓜花叶病毒(CMV)能全部脱去。‘卡萨布兰卡’不定芽培养在固体培养基和液体培养基都能很好形成子球;在含有抗病毒剂(10mg／L DHT)的液体培养基中子球增大显著,LSV脱毒效果理想。无病毒子球移到有防虫网的户外栽培,2年后部分植株被检测出病毒,再感染率因品种而不同,‘卡萨布兰卡’高达73％,麝香百合‘乔治亚’仅17％;无病毒植株‘乔治亚’．香华丽百合和‘卡萨布兰卡’的生长高度要比有病毒植株明显增高。     

百合转基因研究进展

从百合遗传转化受体的建立和百合转基因体系的建立两方面综述了建立百合的遗传转化体系的研究情况,后者包括抗生素的选用、转化再生植株的筛选、共培养时间、农杆菌菌株的选择。同时对近10年来在百合遗传转化方面所作的研究工作进行了综述,包括转报告基因,转抗病、抗虫和抗病毒基因,以及转化技术的研究。提出了转基因百合存在的问题,并就其前景进行了展望。     

Analysis of the meiosis in the F1 hybrids of Longiflorum x Asiatic(LA) of lilies (Lilium) using genomic in situ hybridization

Longiflorum and Asiatic lilies of the genus Lilium of the family Liliaeeae are two important groups of modern lily eultivars. One of the main trends of lily breeding is to realize introgression between these groups. With cut style pollination and embryo rescue, distant hybrids between the two groups have been obtained. However, the F1hybrids are highly sterile or some of them could produce a small number of 2n gametes, and their BC1 progenies are usually triploids. Dutch lily breeders have selected many cultivars from these BC1 progenies based on their variation. It is presumably suggested that such variation could be caused by intergenomic recombination and abnormal meiosis during gamete formation in F1 hybrids of Longiflorum x Asiatic (LA) hybrids in Lilium. Therefore, the meiotic process of ten F1 LA hybrids was cytologically investigated using genomic in situ hybridization and traditional cytological methods in the present research.The results showed that: at metaphase I, the homoeologous chromosome pairing among different F1hybrids ranged from 2.0 to 11.4 bi-valents formed by homoeologous chromosomes per pollen mother cell (PMC), and very few multivalents, and even very few bivalents were formed by two chromosomes within one genome rather than homoeologous chromosomes in some PMCs; at anaphase I, all biva-lents were disjoined and most univalents were divided. Both the disjoined bivalents (half-bivalents) and the divided univalents (sister chromatids) moved to the opposite poles, and then formed two groups of chromosomes; because the two resulting half-bivalents retained their axes in the cell undisturbed, many crossover types, including single crossovers, three strand double crossovers, four strand double crossovers, four strand triple crossovers, and four strand multiple crossovers between the non-sister chromatids in the tetrads of bivalents,were clearly inferred by analyzing the breakpoints on the disjoined bivalents. The present investigation not only explained the reason for sterility of the F1 LA hybrids and the variation of their BC1 progenies, but also provided a new method to analyze crossover types in other F1 interspecific hybrids as well.