The Isolation And Expression Analysis Of Loquat PFP Gene

Loquat fruits flesh is soft and juicy, moderately sweet and sour, and delicious. They are loved bypeople. But the majority of loquat fruits in our country are of lower sugar content and light flavor, whichrestricts the development of the loquats to some extent. PFP is one of the most important regulatoryenzyme in the glycolysis process, adjusting the balance between the sugar and acid. Glycolysis speed ismainly determined by the activity of the enzyme, It is a rate-limited enzyme playing a key role inglycolysis. So the study of the PFP gene function and molecular mechanism of sugar metabolism in theloquat has important significance for improving fruits quality. With Zhejang Agriculture and ForestryUniversity adult loquat trees as materials, this study cloned loquat PFP gene, analyzed its expressionlevel during fruits development and explored its molecular regulation mechanism in loquat fruit sugarmetabolism. The results are as follows:1, This study used RACE technology to get full-length cDNA sequence isolated from loquat. Thelength of this gene is of1876bp, which contains a complete open reading frame of1710bp, encoding569amino acids,88bp in5’UTR area,78bp in3’UTR area, containing a poly (A) tail. The analysisshowed that the encoding protein of the PFP gene has the similarity of more than70%to90%comparing with other plants. The cluster analysis showed that the loquat EjPFPb gene encoding proteinis close to that of ricin, poplar etc., while far from that of tobacco etc.2, Using fluorescence in situ hybridization to clear the distribution and expression abundance ofsubgenotype cell and explore the mechanism action in the cell, and the results showed that thetransferred vector of fusion protein was visible in the cell membrane. Using the real-time RT-PCR toanalyze the dynamic expression of PFP gene in fruit development, and the results showed that twovarieties of loquats at different fruit development stages, the loquat EjPFPb gene had differentexpression levels.3, Finally, using the tobacco genetic transformation system to analyze the function of the EjPFPbgene, and establish a set of efficient tobacco genetic transformation system, in order to explore themolecular mechanism of EjPFPb gene in loquat fruit sugar metabolism and provide a theoretical basisfor exploring the mechanism of the formation of loquat fruit quality.