Identification And Analysis Of Potato Storage Protein Patatin

The potato is the fourth largest food crops in China.It is very important for the continuous development and growth of the potato industry to ensure national food security.Protein is one of the most important components of potato,While the Patatin protein as the most important storage protein,it plays a vital roles in the potato quality.But there are few researches on the identification and gene function studies of potato patatin.Therefore,the aim of this study was to combine mass spectrometry and biochemical analysis to identify the potato StPAT family genes,and silencing patatin gene though the VIGS technology by using potato Desiree as the material,so as to predict the function of genes.The results of this study are as follows:(1)In this experiment,a total of 5597 proteins were identified in potato tubers by mass spectrometry,and 31 proteins were identified in the lipid metabolism pathway associated with plant resistance,of which 9 Patatin were initially identified.(2)To further investigate the potato StPAT family,using the Patatin conserved structural domain to identify the StPAT family.The results revealed that a total of 11 members of the potato StPAT gene family were identified,unevenly distributed on chromosomes 2,5,6,8,9 and 10.Gene structure analysis showed that the members of the potato StPAT gene family could be divided into two groups: Class I and Class II.Class Ⅰ contain six members,and ClassⅡ contain five members.Motif 7 only exist in Class Ⅰ,Motif 8 exist in Class Ⅱ.The number of exons among the members ranged from 2 to 7,the amino acid length from 96 to 466,and the isoelectric point range from 4.96 to 9.01.All family members contained the conserved Patatin structural domain.(3)To further determine the number of StPAT families in potato tubers,we extracted and purified Patatin proteins from potato tubers,and successfully purified potato Patatin with a molecular weight of 40 k Da by ion exchange chromatography and affinity chromatography using a protein purifier AKTA Pure 25,and optimized the purification conditions.The results showed that the best purification results were obtained under the conditions of affinity chromatography Elution buffer 0.2 M methyl-α-D-glucopyranoside,p H 7.4.Finally,the purified patatin was subjected to mass spectrometry analysis and a total of six patatin were identified.(4)Combining the results of protein profiling with the raw letter analysis,and the expressin pattern of the patatin in different tissues of potatoes.Finally selected three key Patatin were special express in potato tubers,they are Patatin 05(LOC102605260),Patatin T5(LOC102602048)and Patatin 02(LOC102601036).To test the functions of the three Patatin-encoding genes were initially validated using VIGS technology.It was found that Patatin gene has the function of regulating tuber number and tuber weight per plant.In this study,we used mass spectrometry and bioconferencing techniques at the proteomic and genomic levels to combine to identify the potato storage protein Patatin and to make preliminary predictions about the function of the gene encoding it,providing a theoretical basis for potato protein quality breeding.