Comparative proteomic analysis of tetraploid black locust (Robinia pseudoacacia L.) cuttings in different phases of adventitious root development

Abstract

Key message Comparative proteome analysis revealed the proteins that are differentially expressed during the processes of tetraploid black locust (Robinia pseudoacacia L.) adventitious root formation, which are involved in carbohydrate binding, energy and metabolism, and stress response. Abstract The tetraploid black locust (Robinia pseudoacacia L.) is an important fast-growing tree used in afforestation and is also a protein-rich feed for poultry. Currently, the primary approach for tetraploid black locust reproduction is through cutting propagation. To identify the specific proteins in the rooting process, including the formation of adventitious root primordium (ARP) and the elongation of adventitious root (EAR), a comparative proteomic analysis of the ARP and EAR of tetraploid black locust was performed to identify related proteins that may be involved in regulating ARP formation and the process of EAR. High-resolution two-dimensional electrophoresis (2-DE) followed by colloidal Coomassie staining and mass spectrometric (MS) analysis was used to identify differentially expressed proteins in ARP and EAR. A total of 84 protein spots showed significant expression changes by 2-DE and were successfully identified by MALDI TOF/ TOF MS/MS. Of these, 29 proteins were differentially expressed during the ARP phase, and the remaining 55 proteins did so in the EAR phase. The majority of identified proteins were classified into functional categories, including carbohydrate binding, energy and metabolism, protein degradation/folding and import, cellular cytoskeleton, hormonerelated and oxidation reactions, stress response, cell proliferation, and transcription regulation. Quantitative real-time PCR analysis was performed for selected proteins and showed that not all of the protein expression levels were consistent with the mRNA levels. This study provides the basis for further functional studies of differentially expressed proteins, which will contribute to the understanding of the biochemical processes in adventitious root primordium formation and the elongation of adventitious root. Abbreviations 2-DE Two-dimensional gel electrophoresis MALDI Matrix assisted laser desorption ionization TOF Time of flight TOF MS Time of flight mass spectrometry MS Mass spectrometry IntroductionTetraploid black locust (Robinia pseudoacacia L.) (TRP), which can be produced by doubling the chromosomes of a diploid black locust (2n 9 2) artificially, is a leguminous Communicated by R. Hampp. Electronic supplementary materialThe online version of this article (

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