Abstract
Leaf vascular system differentiation and venation patterns play a key role in transporting nutrients and maintaining the plant shape, which is an important agronomic trait for improving photosynthetic efficiency. However, there is little knowledge about the regulation of leaf vascular specification and development. Here we utilized the rice midribless mutant (dl2) to investigate the molecular changes in transcriptome and proteome profiles during leaf vascular specification and differentiation. Using isobaric tags for relative and absolute quantification (iTRAQ) with digital gene expression (DGE) techniques, a nearly complete catalog of expressed protein and mRNA was acquired. From the catalog, we reliably identified 3172 proteins and 9,865,230 tags mapped to genes, and subsets of 141 proteins and 98 mRNAs, which were differentially expressed between the dl2 mutant and wild type. The correlation analysis between the abundance of differentially expressed mRNA and DEPs (differentially expressed proteins) revealed numerous discordant changes in mRNA/protein pairs and only a modest correlation was observed, indicative of divergent regulation of transcription and translational processes. The DEPs were analyzed for their involvement in biological processes and metabolic pathways. Up- or down- regulation of some key proteins confirmed that the physiological process of vascular differentiation is an active process. These key proteins included those not previously reported to be associated with vascular differentiation processes, and included proteins that are involved in the spliceosome pathway. Together, our results show that the developmental and physiological process of the leaf vascular system is a thoroughly regulated and complicated process and this work has identified potential targets for genetic modification that could be used to regulate the development of the leaf vasculature.
Highlights
The plant vascular system performs important functions, such as delivery of nutrients to the various plant tissues, supply of mechanical support, as well as acting as an effective long-distance communication system relating to abiotic and biotic conditions above and below ground (Lucas et al, 2013)
In an attempt to have a global view of gene activity in the process of leaf vascular differentiation in rice, and probe candidate genes involved in plant vascular differentiation, we examined the abundance of transcripts and proteins by digital gene expression (DGE) and isobaric tags for relative and absolute quantification (iTRAQ) technology, respectively, and correlated the abundance difference between mRNAs and proteins
Protein Identification and Quantification Using iTRAQ To investigate the proteome expression differences between wild type (WT) and MT, proteins from two biological replicates per phenotype were analyzed with iTRAQ technology
Summary
The plant vascular system performs important functions, such as delivery of nutrients to the various plant tissues, supply of mechanical support, as well as acting as an effective long-distance communication system relating to abiotic and biotic conditions above and below ground (Lucas et al, 2013). Plant leaf vascular differentiation and venation patterns play a key role in determining plant shape, which is an important agronomic trait. Considerable effort has been made examining genetic control of vascular differentiation using vascular system mutants, with several factors identified that affect developmental and physiological processes of the plant vascular system (Mahonen et al, 2000; Dettmer et al, 2009; Ohashi-Ito and Fukuda, 2010). In spite of extensive descriptions about vascular development and patterning, the molecular and physiological mechanisms governing the strictly regulated process remain to be elucidated
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