| Sugar beet rhizomania is a serious disease affecting sugar beet production,which can lead to a sharp decrease in sugar beet yield and sugar content.In this study,the mechanism of resistance to rhizomania was studied for the first time in China,but no related reports have been reported before.In this paper,76 sugar beet breeding materials were tested for identification of resistance to rhizomania,three different protein extraction methods were compared,and the conditions of sugar beet two-dimensional electrophoresis system were optimized.The changes of enzyme activity of resistant enzymes in different stages of virus stress treatment and untreated materials were compared.After completing two dimensional electrophoresis,protein gel spots with obvious difference between disease-resistant and susceptible materials were removed,and the protein in glue was identified and analyzed by Hua Da Gene Company.The findings are as follows:(1)Five resistant materials with good resistance to rhizomania were screened out from different breeding materials in field identification experiment.Among them,HBI-1 and N98122 are high resistance materials.HBX-5 and IX-1 were selected as high sensitive materials.They were used as disease-resistant and susceptible materials in subsequent tests.Among the imported materials,high sugar type materials and standard materials,3,2 and 2 materials with good performance were screened respectively.(2)Optimum conditions of beet two-dimensional electrophoretic system: TCA/ acetone method is more effective in three kinds of protein extracting in beet leaves.Two-dimensional Electrophoresis of Sugar beet leaves in pH range of 4 ~ 7 for IPG strips is better.The effect of 7cm IPG film on 150 μ g sample is better.The effect of 17 cm IPG film on 300 μ g sample was better.(3)The peak date of enzyme activity after stress reaction was not the same,and the peak date appeared at the seedling age of 45d(initial stage of infection),and the peak date of peroxidase was 50 days at seedling age,and the speed of gene expression in stress response was not the same.(4)The results of field yield and quality identification of susceptible materials showed that the sugar content of the resistant materials were all higher than that of the control,and the anti-1(HBI-1)’ yield of root per mu was slightly lower than that of the control,and the anti-2(N98122)was higher than that of the control.Sugar production was higher than that of control.The comprehensive performance of anti-1 HBI-1 was better than that of anti-2 N98122.The root yield and sugar content of susceptible materials were lower than that of control.(5)After two-dimensional electrophoresis,a total of 169 proteins were detected after deducting 78 distinct protein spots.Among them,25 proteins were detected only in susceptible materials and 111 proteins were detected only in resistant materials.At the same time,13 proteins were detected in anti 1 and anti 2 samples,28 proteins were detected in anti 1 sample and 70 proteins were detected in anti 2 samples.Through gene annotation and pathway analysis,72 KOs and 114 pathways were annotated.145 proteins were annotated to 208 go annotations.134 of them could be annotated on NR database.Through comparing the database,the main functional proteins expressed in resistant materials are photosynthesis,glucose metabolism,respiratory antioxidation,signal transduction,lipid metabolism and some unknown proteins,while susceptible materials mainly express photosynthesis proteins and respiratory antioxidant proteins.This is the protein expressed in the stress response.A protein sequence,Bv7171670 jfwy.t1 was found in the anti-2 material.It was inferred that the protein At1g52660 might be related to plant disease resistance. |
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