Differential transcriptional networks associated with key phases of ingrowth wall construction in trans-differentiating epidermal transfer cells of Vicia faba cotyledons.

Hui-Ming Zhang,Hans Weber,Ruslana Radchuk,Xue Xia,John W Patrick,Simon Wheeler,Christina E Offler

doi:10.1186/s12870-015-0486-5

Abstract

BackgroundTransfer cells are characterized by intricate ingrowth walls, comprising an uniform wall upon which wall ingrowths are deposited. The ingrowth wall forms a scaffold to support an amplified plasma membrane surface area enriched in membrane transporters that collectively confers transfer cells with an enhanced capacity for membrane transport at bottlenecks for apo-/symplasmic exchange of nutrients. However, the underlying molecular mechanisms regulating polarized construction of the ingrowth wall and membrane transporter profile are poorly understood.ResultsAn RNAseq study of an inducible epidermal transfer cell system in cultured Vicia faba cotyledons identified transfer cell specific transcriptomes associated with uniform wall and wall ingrowth deposition. All functional groups of genes examined were expressed before and following transition to a transfer cell fate. What changed were the isoform profiles of expressed genes within functional groups. Genes encoding ethylene and Ca2+ signal generation and transduction pathways were enriched during uniform wall construction. Auxin-and reactive oxygen species-related genes dominated during wall ingrowth formation and ABA genes were evenly expressed across ingrowth wall construction. Expression of genes encoding kinesins, formins and villins was consistent with reorganization of cytoskeletal components. Uniform wall and wall ingrowth specific expression of exocyst complex components and SNAREs suggested specific patterns of exocytosis while dynamin mediated endocytotic activity was consistent with establishing wall ingrowth loci. Key regulatory genes of biosynthetic pathways for sphingolipids and sterols were expressed across ingrowth wall construction. Transfer cell specific expression of cellulose synthases was absent. Rather xyloglucan, xylan and pectin biosynthetic genes were selectively expressed during uniform wall construction. More striking was expression of genes encoding enzymes for re-modelling/degradation of cellulose, xyloglucans, pectins and callose. Extensins dominated the cohort of expressed wall structural proteins and particularly so across wall ingrowth development. Ion transporters were selectively expressed throughout ingrowth wall development along with organic nitrogen transporters and a large group of ABC transporters. Sugar transporters were less represented.ConclusionsPathways regulating signalling and intracellular organization were fine tuned whilst cell wall construction and membrane transporter profiles were altered substantially upon transiting to a transfer cell fate. Each phase of ingrowth wall construction was linked with unique cohorts of expressed genes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0486-5) contains supplementary material, which is available to authorized users.

Highlights

Transfer cells are characterized by intricate ingrowth walls, comprising an uniform wall upon which wall ingrowths are deposited
126,200,279 high-quality reads, in which 97% of nucleotides have a Phred quality score of ≥ Q20 level (Table 1) were de novo assembled into 131,279 contigs (>200 bp) with a N50 value of 980 bp and an average length of 423 bp
Clustering yielded 74,659 unigenes with 33,902 consensus sequences grouped into 11,083 distinct clusters and 41,567 singletons (Table 1)

Summary

Introduction

Transfer cells are characterized by intricate ingrowth walls, comprising an uniform wall upon which wall ingrowths are deposited. The ingrowth wall forms a scaffold to support an amplified plasma membrane surface area enriched in membrane transporters that collectively confers transfer cells with an enhanced capacity for membrane transport at bottlenecks for apo-/symplasmic exchange of nutrients. Flange ingrowths form as ribs or bands of wall material while the more commonly occurring reticulate wall ingrowths arise as numerous wall papillae that develop at right angles to the original wall. The extent of their reticulation varies from cylindrical papillae alone to ones in which papillae branch and fuse to generate fenestrated wall layers [4]. The uniform wall and reticulate wall ingrowths form the ingrowth wall [1]

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