Effects Of Dimethyl Sulphonate On Proteome Of Paulownia Seedlings Infected By Phytoplasma

Paulownia has a relatively large economic value.It plays an important role in improving the ecological environment and improving the living standards of farmers.Paulownia is susceptible to phytoplasma invasion and suffers witches' broom disease.The disease damage is extremely serious,and it is extremely harmful to forestry,economic,environment and other aspects caused huge losses.In order to effectively prevent and control this disease,although many studies have been done at present,the pathogenesis of the disease remains unclear.In order to gain insights into the protein changes during the Paulownia witches' broom disease occuring,isotopic relative and absolute quantification(i TRAQ)techniques were used to identify healthy seedlings,witches' broom seedlings and different concentrations(15 mg·L-After treatment with 1 and 75 mg·L-1 dimethylsulfate(DMS),the protein of diseased seedlings with healthy morphology was recovered,and functional annotation and differential expression analysis were performed.The following results were obtained:(1)A total of 2969 proteins were identified in 4 samples of Paulownia fortunei.There were 2 959,2 692,and 2 928 proteins annotated to the NCBI protein database,the Swiss-Prot database,and the tr EMBL database.According to the results of functional classification,there were a total of 2 640 protein alignmentss in the GO database,which were annotated to 51 functional categories.There have 2 581 protein alignments were mapped to the COG database and were divided into 23 functional categories.Among the KEGG database alignments,2 324 proteins were involved in 121 metabolism pathways,mainly involved in metabolic pathways,biosynthesis of secondary metabolism,and photosynthesis.A total of 356 differentially expressed proteins were found in the paulownia group,which were mainly involved in metabolic pathways,secondary metabolism synthesis,photosynthesis,and photosynthetic carbon fixation pathways.(2)A total of 2628 proteins were identified in 4 samples of P.tomentosa.And 2 615,2,366,and 2586 proteins were respectively annotated to the NCBI protein database,the Swiss-Prot database,and the tr EMBL database,.The functional classification results showed that a total of 2 327 proteins were annotated into the GO database and divided into 50 functional categories.2 259 protein alignments were compared to the COG database and annotated to 23 functional categories.The results of the KEEG database showed that a total of 2 049 proteins were involved in 119 metabolic pathways of which mainly involved in metabolic pathways,synthesis of secondary metabolites,and photosynthetic pathways.A total of 324 differential proteins were obtained from the P.tomentosa sample alignment groups and mainly involved in metabolic pathways,secondary metabolite synthesis,photosynthesis,and photosynthesis carbon fixation pathways.(3)Differential protein screening was performed on Paulownia fortunei and Paulownia tomentosa.The lateral differences of the proteins identified by Paulownia were compared and analyzed.Finally,five proteins,ribosomal protein L11 and malate dehydrogenase(MDH),Photosystem II captures chlorophyll a/b binding protein,photosystem I subunit II,calmodulin(Ca M),and an unknown function protein,were found.Mainly proteins involved in photosynthesis,photosynthetic bio-carbon fixation,and ribosome and other biological metabolic pathways.Analysis of the functions involved in the discovery of light-system II light-harvesting chlorophyll a/b-binding protein and photosystem I subunit II protein contributes to the regulation of photosynthesis in plants by affecting the synthesis of light-harvesting complexes(LHC)and thus in the process of Paulownia induction plays a regulatory role;malate dehydrogenase(MDH)affects the water transport and NADH and ATP synthesis in plants,and shows some disease resistance by regulating the metabolism of energy and substances in plants;Calmodulin(Ca M)plays an important role in stimulating the autoimmune response of plants in interaction of plants and pathogens;ribosomal protein L11 participates in the specific interaction between pathogen and host during paulownia infection of phytoplasma.(4)Four proteins were randomly selected from the differentially expressed proteins of Paulownia tomentosa and Paulownia fortunei by q RT-PCR.The results showed that there were 3 proteins at the transcription level and protein expression in each of Paulownia tomentosa and Paulownia fortunei.The level showed a consistent trend,and the trend of one protein expression was inconsistent.The reasons for this inconsistency may be related to m RNA degradation,protein degradation,and protein folding modifications.The results of this study have identified the proteins involved in the occurrence of witches' broom and the physiological and biochemical pathways involved in them,providing a reference basis for understanding the pathogenesis of witches' broom and clarifying the occurrence of Paulownia ssp and further research on the pathogenesis of witches' broom.