Arabidopsis Response to Inhibitor of Cytokinin Degradation INCYDE: Modulations of Cytokinin Signaling and Plant Proteome

Cytokinins are multifaceted plant hormones that play crucial roles in plant interactions with the environment. Modulations in cytokinin metabolism and signaling have been successfully used for elevating plant tolerance to biotic and abiotic stressors. Here, we analyzed Arabidopsis thaliana response to INhibitor of CYtokinin DEgradation (INCYDE), a potent inhibitor of cytokinin dehydrogenase. We found that at low nanomolar concentration, the effect of INCYCE on seedling growth and development was not significantly different from that of trans-Zeatin treatment. However, an alteration in the spatial distribution of cytokinin signaling was found at low micromolar concentrations, and proteomics analysis revealed a significant impact on the molecular level. An in-depth proteome analysis of an early (24 h) response and a dose-dependent response after 168 h highlighted the effects on primary and secondary metabolism, including alterations in ribosomal subunits, RNA metabolism, modulations of proteins associated with chromatin, and the flavonoid and phenylpropanoid biosynthetic pathway. The observed attenuation in stress-response mechanisms, including abscisic acid signaling and the metabolism of jasmonates, could explain previously reported positive effects of INCYDE under mild stress conditions.

Comparative Analysis of Transcriptomes to Identify Genes Associated with Fruit Size in the Early Stage of Fruit Development in Pyrus pyrifolia

Pear (Pyrus L.) is an important commercial fruit in the world. The fruit size is one of the important characters in fruit quality. The previous research reported that the fruit size of pear was mainly caused by the number of cell in about 40 days after blossom (DAB) in nature. However, studies about the mechanisms underlying cell division in young fruit development are very limited in pear. Two pear accessions codenamed ‘GH59B’ with big fruit and ‘GH81S’ with small fruit in three stages were sampled and the RNA-seq high-throughput sequencing was used to evaluate changes of gene transcription levels in the early stage of fruit development. The difference of cell size among two samples was little in 40 DAB, implying that the difference of the fruit size was caused by the number of the cell. More than 274,517,982 high quality reads from six libraries of fruit development were sequenced. A total of 797 differentially expressed genes (DEGs) were identified. Three cytokinin dehydrogenase genes and two gibberellin 2-beta-dioxygenase gene were identified in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related to zeatin and gibberellin. Their expression was upregulated at 20 DAB in ‘GH81S’ and at 30 DAB in ‘GH59B’, suggesting that the small fruit size might be related to the early degradation of cytokinin and gibberellin inducing a short period of cell division. A total of 38 DEGs of transcription factors were found and 23 DEGs including NAC, ERF and bHLH transcription factors were highly related with cytokinin dehydrogenase and gibberellin dioxygenase genes. Altogether, the results of the present study provide information from a comprehensive gene expression analysis and insight into the molecular mechanism underlying the difference of fruit size in Pyrus pyrifolia.