engleski

Plastid stromal proteome of in vitro grown horseradish (Armoracia lapathifolia Gilib.) plantlet, tumor and teratoma tissues

Crown-gall tumors were induced on horseradish leaf fragments with a wild strain B6S3 of A. tumefaciens. During subcultivation, two different transformed phenotypes were established: unorganized tumor-TN and teratoma-TM, composed of malformed shoots. The study has been focused on plastids. During tumor transformation, these organelles change from developed chloroplast to juvenile proplastide stage. Stromal proteome of tumor plastids and chloroplasts from leaf cells were compared. Intact plastids were lysed under non-denaturating conditions and stromal proteins were digested with trypsin. Triptic peptides were separated by C18 nano-chromatography and measured online with LTQ-Orbitrap Discovery MS. Peak lists (generated by MaxQuant) were used by X!Tandem to search A. thaliana proteome -TAIR database. 321 leaf proteins, 70 TM and 54 TN proteins were identified. They were classified according to MapMan Bin classification, reported in PPDB. Leaf proteins are involved in photosynthesis, major/minor CHO, amino acid, nucleotide and RNA metabolism, glycolysis, OPPP, organic transformation, lipids, hormone, isoprene, tetrapyrrole, protein synthesis and redox regulation. TM proteins were missing in minor CHO and RNA metabolism, isoprene synthesis. TN proteins were missing in minor CHO and nucleotide metabolism, lipid, hormone, isoprene and tetrapyrrole synthesis. Our results show that cell transformation and tissue developmental stage influence the composition of stromal proteome.

plastids; stromal proteome; differentiation; tumor transformation

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