枇杷脂类相关质体蛋白基因PAP的克隆及表达分析

本研究以‘大五星’(红肉)和‘川农159’(白肉)枇杷不同发育阶段的果皮、果肉为试验材料,采用同源克隆法从枇杷中分离PAP基因后采用qRT-PCR对两种材料不同时期的皮和肉进行定量分析。结果如下:该基因的CDS区具有一个长744 bp的开放阅读框,编码247个氨基酸。氨基酸序列的同源性分析表明,枇杷PAP蛋白与白梨(Pyrus bretschneideri)相似性高达97%。进化树也进一步说明,枇杷与白梨的进化关系最近。定量分析表明:白肉枇杷果肉中PAP基因的表达量平均水平较低,成熟的果皮果肉中PAP基因的表达量都表现为红肉枇杷显著高于白肉枇杷,表明PAP影响枇杷色素形成。本试验结果更清晰地呈现出了EjPAP基因的序列结构以及PAPs蛋白在枇杷果实呈色过程中的重要作用。

Cloning and Expression Analysis of Plastid Lipid-associated Proteins Gene PAP in Eriobotrya japonica Lindl.

In this study,the PAP gene was isolated from loquat(Eriobotrya japonica)by RT-PCR and homologous cloning used the peel and pulp of’Dawuxing’(red flesh)and’Chuannong 159’(white flesh)at different developmental stages,and then the skin and flesh of the two materials at different stages were quantitatively analyzed by qRT-PCR.The results were as follows:The CDS region of the gene was 744 bp long,with an open reading frame744 bp long,encoding 247 amino acids.Homology analysis showed that loquat PAP protein had high similarity97%with other plant PAP proteins,among which it had the highest similarity with white pear(Pyrus bretschneideri)protein.The evolutionary tree analysis further indicated that the evolutionary relationship between loquat and white pear was the most recent.The results of qRT-PCR analysis showed that the average expression level of PAP gene in red loquat pulp was higher than that in white loquat pulp,indicated that PAP affected the formation of loquat pigment.The results of this study clearly showed the sequence structure of EjPAP gene and the important role of PAPs protein in the process of loquat fruit color.