Research And Analysis Of Differential Proteins From Tomato Yellow Leaf Mutant

Tomatoes are important vegetable crops within the scope of the world. Its economic value and application value also gets more and more extensive attention. Leaf color mutant is one of the life phenomenon in the plant kingdom. Many kinds of leaf color mutants including the mutant with nv gene and the the mutant with dg gene have been found in tomatoes. The research of these mutants not only reveals the process of chlorophyll synthesis and degradation and theory about chloroplasts genetic and development but also provides the foundation for the application of leaf color mutant. The tomato institute of Northeast Agricultural University names the natural leaf color mutant which was found in Zhongshusihao group as06883. This mutant is different from other known leaf color mutants. The study to this mutant not only directly reveals the process of chlorophyll synthetic metabolic but also play a positive role in the clarification to the system built of Photosynthetic membrane and photosynthetic function.expression and regulation of genes about Chloroplasts development and the form built of the environmental signal to chloroplasts and other organs.Tomato yellow leaf mutant06883which has been found from tomato inbred lines06884is a kind of nature leaf mutant. The research firstly selects the most suitable protein extraction method and two-dimensional electrophoresis (2-DE) system for the proteomics analysis of tomato yellow leaf mutant06883.And differences proteomic analysis of06883and06884leaves which aims at the study the variation laws of total06883leaves proteins provide more theoretical basis for the leaf color mutant mechanism. The research results show:1. Tomato yellow leaf mutant06883protein extraction methods has been screened.The proteins of the fresh leaves of06883selected as experiment materials have been extracted with the method of TCA/acetone precipitation, Tris-base precipitation and Tris-HCl separately. The extraction effect has been compared between these three methods in protein yield, single-dimensional and two-dimensional electrophoresis map. The results show that the improved TCA/acetone precipitation method is the best method of protein extraction.2. The two-dimensional electrophoresis (2-DE) system of tomato yellow leaf mutant06883has been set up.The reasearch selects the best conditions from pH range of immobilized gradient strips(IPG), the quantity of sample-loading and dyeing way. The results show that24cm pH4-7IPG is suitable for leave’protein separation of tomato yellow leaf mutant06883,1200μ g is best quantity of protein sample-loading, R350is the most suitable way of dyeing.3.The results show the expressions of twenty-four protein points have changed significantly through qualitative and quantitative analysis. Two of these protein points only exist in tomato yellow leaf mutant06883and three of these protein points only exist in Zhongshusihao06884. The expressions of twelve protein points have increased and the expressions of twelve protein points have decreased from tomato yellow leaf mutant comparing with the expressions of control group Zhongshusihao06884(P<0.05). Twenty-three protein points identified successfully from twenty-four differential expression protein points through Mass spectrometry have been classified results show that these differential expression proteins respectively belong to energy related proteins, metabolic related proteins, combined with protein, signal transduction protein, resistance related proteins, etc. The identification of these differential expression proteins provides theoretical basis for the further understanding of tomato yellow leaf mutants’ molecular and mutations mechanism.