【作者】 李凤兰；

【导师】 胡宝忠；

【作者基本信息】 东北农业大学 ， 植物学， 2004， 硕士

【摘要】 本研究主要是通过对不同花色一串红的花发育过程、组织培养快繁方法和Ss5MaT1基因进行了研究，首次观察一串红的花芽分化和胚胎学有关内容，探讨了影响一串红结实性的原因、最佳组织培养快繁体系，揭示了花色基因Ss5MaT1的多态性， 花发育研究结果表明，一串红的花芽分化是从第4个叶节开始，在第9个叶节时分化结束，花药壁的发育方式为双子叶型，成熟花粉粒为三细胞型，胚囊的发育方式为蓼型，表明一串红结实率低的原因不是由于雄、雌配子体的发育不良所造成。八种花色的一串红的结实率、落花率及成熟胚珠败育率调查结果表明一串红的平均结实率为15.81%，平均落花率为23.13%，成熟胚珠败育率为84.13%。 一串红组织培养的最佳外植体为带腋芽的茎段，茎段的芽丛诱导的最佳培养基为1/2MS+NAA0.1mg/L+6-BA2.0mg/L+IBA 0.5mg/L，其分化率为71.4%，芽增殖率为140．6%；最佳生根培养基为1/4MS+NAA 0.5mg/L,生根率为91.6%，初步建立了快繁体系。通过RT-PCR方法获得了八种一串红中Ss5MaT1基因的916bp的cDNA片段，根据获得的cDNA片段设计了4对引物，采用嵌套式PCR的方法，以RT-PCR的产物为模板进行扩增，对扩增产物进行SSCP分析，发现只有一对引物的扩增产物出现了差异带，而其它三对引物无差异带。基因测序的结果表明：RT-PCR扩增获得的cDNA片段，同源性相当高，只在鲑鱼肉色花和玫瑰红色花两种花色中发现了一个点突变，即玫瑰红色花在744C T，鲑鱼肉色花中的678C T，玫瑰红色花中的点突变导致异亮氨酸（Ile）突变为苏氨酸（Thr），而鲑鱼肉色花中的点突变导致了缬氨酸（Val）突变成丙氨酸（Ala），而其它花色没有发现突变点。基因克隆测序的结果与PCR-SSCP分析的结果一致，表明PCR-SSCP可以用于花色基因的突变点的检测。更多还原

【Abstract】 In this study,I determined the cause of affecting fecundity of salvia splendens、the best system of tissue culture fast propagations system and clarified flower color gene Ss5MaT1 polymorphism, through studying on the embryological structure in flower delevoping, the way of tissue culture fast propagations and flower color gene by PCR-SSCP combining with sequencing .The result of flower delevoping showed that the flower bud morphological differentiation began at 4th leaf burl and finished at 9th leaf burl, and the wall of anther was dicotyls type. Moreover, mature microspore was three-cell type, and the delevoping payway of megaspore was polygonum type, which testified that the seeding rate low wasn’t due to transmissibility male gametophyte and female gametophyte failing growth.The results of observating the number of seedings in different color breed indicated the averge seeding rate in Saliva splendens was 15.81%, and the averge drop flower rate was 23.13%. the mature ovule failing growth rate was 84.1%.The best explant in the culture was stem segment with bud, and the best medium of buds inducing was 1/2MS+NAA0.1mg/L+6-BA2.0mg/L+IBA 0.5mg/L, differentiation rate was 71.4%, buds multiplication rate was 140.62%. The best of rooting medium was 1/4MS+NAA 0.5mg/L, taking root rate was 91.6%.According to the sequences of Ss5MaTI in GenBank, I obtained 916bp Ss5MaT1gene partial cDNA sequence of eigh species by RT-PCR.According to cDNA sequence that obtained in RT-PCR, four pairs of primers were designed. Aim segment was amplificated throgh nested PCR. PCR-SSCP technology was applied to detect SNPs of the Ss5MaT1. Detected results demonstrate that only one pair primer amplification segment had polymorphism, but others had not been different. The result of sequening showed that the cDNA segment that was amlificated by RT-PCR had high homology, only the Salmon and the rose had a mutation point respectively. Rose: 744 C T, Ile Thr; Salmon: 678 C T, Val Ala, and others had not mutation point. The cloning and sequening result was identical to the result of PCR-SSCP, which showed PCR-SSCP could examine the mutation of flower gene. Master Candidate：Li Fenglang <WP=10>Speciality: botany Advisor: Professor Hu Baozhong更多还原