A method for quantitative screening of stigma differential proteins in strawberry by using dda-dia alternate collection

Abstract

The invention discloses a method for using DDA-DIA to alternately collect and quantitatively screen differential proteins in the stigma of red-cheeked strawberry, comprising the following steps: 1), establishing a peptide information library of stigma-cap protein in DDA-collecting mode; 2), collecting in DIA mode Sample information to be tested: take red cheek strawberry stigmas at the small-bud stage, middle-bud stage, and big-bud stage, prepare peptide solutions respectively, and then use DIA mode to collect quantitative data; 3), screening differential proteins: obtain from step 1) The information library list and the DIA original file obtained in step 2) were imported into the analysis software Spectronaut Pulsar for matching quantification and T test analysis, and the samples at the middle bud stage were used as the total control to screen out p<0.05 and the samples at the small bud stage and the samples at the large bud stage were respectively Differential proteins with an expression fold ≥ 1.5 or ≤ 2 / 3 compared to samples at the mid-bud stage. Using this method, the quantitative screening of differential proteins in the stigma of strawberry can be realized.

Description

technical field [0001] The invention relates to the field of plant proteomics, and provides a method for quantitatively screening stigma differential proteins of strawberry. Background technique [0002] Red cheek is one of the main strawberry varieties with a wide area in my country, and it is also one of the most frequently used parent materials in conventional cross breeding. , which is of great significance to increase the success rate of crossbreeding strawberry hybridization, obtain more hybrid offspring, and improve hybridization efficiency. During the development of organisms, proteins play an important role as the execution and composition of life activities. Studying the proteomics of the stigma of strawberry and screening the protein differences in each stage of the stigma will help explain the mechanism of its receptivity change, and provide information for breeding practice. theoretical basis. [0003] Proteomics was proposed in 1994. After entering the 21st ce...

AI Technical Summary

Problems solved by technology

Among them, PRM and MRM are mainly technical means for targeted quantification, which are suitable for the verification of differential proteins after quantitative screening; DDA is currently the most widely used quantitative technology for proteomics, and its acquisition method is to select the abundance of mass spectrometry based on real-time primary mass spectrometry The highest 20 or 30 precursor ions are analyzed by secondary mass spectrometry. This method has high collection efficiency and simple experimental design. However, due to the characteristics of real-time determination of the collection sequence, the repeatability is poor, and low-abundance proteins are not easy to collect; DIA is the most popular method in recent years. Newly developed quantitative technology, the acquisition method is to divide the scanning range into several mass-to-charge ratio (m / z) windows, and all precursor ions in each window are analyzed by secondary mass spectrometry. This method has high data reproducibility, The secondary spectrum is complete and accurate in quantification, but it needs to establish a correlation spectrum library of precursor ions and product ions in advance, and the experimental design is more complicated

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