【作者】 陈芳芳；

【导师】 黄坚钦； 黄有军；

【作者基本信息】 浙江农林大学 ， 森林培育， 2010， 硕士

【摘要】 山核桃(Carya cathayensis Sarg.)是我国特有的名优干果和木本油料植物,具有很高的经济价值。山核桃属雌雄同株异花木本植物,存在童期长、雌雄花发育时间不一致的问题,直接影响山核桃的产量。本论文通过克隆若干山核桃成花基因并探讨其表达模式,为研究山核桃成花机理并从根本上提高山核桃产量奠定一定基础。本论文将利用同源克隆及基因组步移技术获得山核桃若干成花相关基因,通过Real-Time RT PCR技术和RNA原位杂交技术分析成花相关基因的时空表达情况。主要研究结论如下:1、获得FT、AG和AP1同源基因的片段,分别命名为CcFT、CcAG和CcAP1。对CcFT、CcAG和CcAP1基因的序列分析表明它与其他植物相应的同源基因都有极高的同源性,这种结构上的相似性使笔者有理由推测它们在功能上的相似性。2、对CcAP1基因的表达分析发现,该基因在整个雌花芽形成过程中的表达情况并没有呈现明显的规律,多数时期表达量均很低,或几乎不表达。相关研究显示AP1与萼片和花瓣的发育有关,山核桃雌花花被缺失,这可能是造成CcAP1基因表达量低的原因之一。3、CcLFY基因在整个雌花芽形成过程中均有表达,在不同时期间表达量差异很大。Real-Time RT PCR实验结果显示,在3月中上旬CcLFY的表达量比之后的表达量明显要高,在而之后各样品的相对表达量相对较低。同时还发现CcLFY的表达量受到温度的影响,低温在一定程度上会降低CcLFY的表达量。4、原位杂交实验结果显示,雌花芽原基时期的CcLFY杂交信号不是很强烈,随着花序原基顶端生长锥逐渐变平,杂交信号有所增强,之后形成小花、小苞片过程中,均有CcLFY检测信号,这些结果说明CcLFY在山核桃雌花芽的形成中具有重要作用,并可能影响雌花芽形成后的花器官分化。更多还原

【Abstract】 Carya cathayensis Sarg. is endemic to Chinese famous dried fruit and Woody oil plant, with high economic value. Carya cathayensis is a hermaphrodite outcrossing woody plants, have problems such as a long time ahead of maturation and male and female flower development time inconsistency which directly affect it’s production. This paper will probe into Carya cathayensis flowering mechanism and the mechanism of sex floral organs, for fundamentally improving the Carya cathayensis production or realization of precocious and dwarf provide the scientific basis. The paper will use homology cloning and genome walking techniques clone a number of flower-related genes in Carya cathayensis, use Real-Time RT PCR technology and RNA in situ hybridization analysis gene expression in different flowering time and different issues of female flower bud. The main conclusions are as follows:FT, AG and AP1 homologous gene fragments have been cloned in Carya cathayensis, named CcFT, CcAG and CcAP1. Homology analysis between CcFT, CcAG and CcAP1 gene sequence and the corresponding genes of other plants showed that they have a high homology,this structural similarity suggested that they have similar function.Expression analysis of the CcAP1,we found that its expressing during the female flower bud formation did not show obvious rule.Most times the expression were low or almost no expression. Related studies have shown that AP1 may related to development of the sepals and petals. Carya cathayensis female flower lack of perianth,that may result in low expression of CcAP1.Carya cathayensis CcLFY expresses during the whole process of the formation of the female flower bud , but very different in different times. Real-Time RT PCR results showed that, CcLFY expresses significantly higher in 3 months early than all samples following, expression of each late sample is relatively low. Also found that the expression of CcLFY affected by temperature, low temperature will reduce the expression of CcLFY to some extent.In situ hybridization results showed that, CcLFY hybridization signal during female flower bud primordia is not very strong, with the growth cone in the top inflorescence primordia gradually flattened, hybridization signal was enhanced. CcLFY hybridization signals also detected during the formation of small flowers, small bracts process. These results suggest that CcLFY may plays an important role in female flower bud formation in Carya cathayensis, and may also affect the formation of organ differentiation.更多还原