【作者】 郑杨；

【导师】 陈晓流； 陈学森；

【作者基本信息】 山东农业大学 ， 果树学， 2008， 硕士

【摘要】 为分离自交亲和的四倍体植物中国樱桃(Prunus. psedocerasus L.)花粉S-决定子基因,本研究采用中国樱桃品种‘泰小红樱’为试材,根据李属不同树种的花粉SFB基因序列设计兼并引物,应用RT-PCR、RACE等技术分离并鉴定了几个花粉SFB基因。结果如下:1应用RT-PCR技术,从花粉cDNA中分离到了1个中间片段,与扁桃花粉SFB基因高度同源。用3’-RACE分离到1个3’末端序列,与扁桃花粉SFB基因高度同源。分离到3个5’末端序列,分别与扁桃和梅的花粉SFB基因高度同源。2拼接5’末端序列、RT-PCR中间片段和3’末端序列后,得到了一个全长序列,全长1171bp,开放式阅读框编码375个氨基酸,N端氨基酸序列具有花粉SFB基因的F-box基序,与扁桃花粉SFB基因高度同源,并具备李属花粉SFB基因的序列特征,命名为SFBc,在GenBank上的注册号为EU267943。3另外两个5’-端序列核酸序列长度均为656bp,编码214个氨基酸,其N端氨基酸序列都具有花粉SFB基因的F-box基序,分别与梅和扁桃的花粉SFB基因高度同源,分别命名为SFBa和SFBb。SFBa和SFBb在GenBank上的注册号分别为EU267941和EU267942。4基因的进化树表明,中国樱桃花粉SFB基因与李属花粉SFB/SLF位于同一个进化分枝,而苹果属花粉SFB基因与茄科、玄参科的处于另外一个进化枝。更多还原

【Abstract】 For isolating the pollen S-determinant genes in Chinese cherry (Prunus. psedocerasus L.), which is a self-compatibility tetroploid species, an Chinese cherry cultivar Taixiaohongying was used as metierial. Using degenerate primers designed based on the conserved regions of the reported SLF/SFB genes, several different cDNA clone were isolated by RT-PCR, The results are as follow:1. One middle segment cDNA clone was isolated from pollen by cDNA RT-PCR., which chosely homologous to those from amond. 3’RACE in combination with RT-PCR allowed isolation of one 3’segment, which chosely homologous to those from amond. 5’RACE combination with RT-PCR allowed isolation of three 5’segments, which chosely homologous to those from amond and Japanese apricot.2. The middle segment, 3’segment and one of the three 5’segment composed one full-length cDNA , length 1171bp. The deduced ORFs encode a F-box protein, length 375, which chosely homologous to those from amond, named SFB. We conclude that SFBc (EU267943) is the pollen SFB genes in Chinese cherry.3. The other two 5’segments, length 656bp, whose corresponding gene SFBa and SBFb were specifically expressed in pollen in an S-haplotype specific manner. The deduced ORFs of the the genes encode twe divergent F-box protein. We conclude that SFBa (GeneBank accession number: EU267941) and SFBb (EU267942) are the pollen SFB genes in Chinese cherry.4. The phylogenetic trees for SFB proteins reported in Solanaceae, Scrophulariaceae and Rosaceae were constructed. It was found that SFB from Chinese cherry were more closely related to those from Prunus (genus of Rosaceae) than those from Malus (genus of Rosaceae), Petunia (a genus of Solanaceae) and Antirrhinum (genus of Scrophulariaceae).更多还原