The Cell Differentiation of Idioblast Myrosin Cells: Similarities With Vascular and Guard Cells

Idioblasts are defined by abnormal shapes, sizes, and contents that are different from neighboring cells. Myrosin cells are Brassicales-specific idioblasts and accumulate a large amount of thioglucoside glucohydrolases (TGGs, also known as myrosinases) in their vacuoles. Myrosinases convert their substrates, glucosinolates, into toxic compounds when herbivories and pests attack plants. In this review, we highlight the similarities and differences between myrosin cells and vascular cells/guard cells (GCs) because myrosin cells are distributed along vascular cells, especially the phloem parenchyma, and myrosin cells share the master transcription factor FAMA with GCs for their cell differentiation. In addition, we analyzed the overlap of cell type-specific genes between myrosin cells and GCs by using published single-cell transcriptomics (scRNA-seq) data, suggesting significant similarities in the gene expression patterns of these two specialized cells.

Asymmetric wall ingrowth deposition in Arabidopsis phloem parenchyma transfer cells is tightly associated with sieve elements

In Arabidopsis, polarized deposition of wall ingrowths in phloem parenchyma (PP) transfer cells (TCs) occurs adjacent to cells of the sieve element/companion cell (SE/CC) complex. However, the spatial relationships between these different cell types in minor veins, where phloem loading occurs, are poorly understood. PP TC development and wall ingrowth localization were compared to other phloem cells in leaves of Col-0 and the transgenic lines AtSUC2::AtSTP9-GFP and AtSWEET11::AtSWEET11-GFP that identify CCs and PP respectively. The development of PP TCs in minor veins, indicated by deposition of wall ingrowths, proceeded basipetally in leaves. However, not all PP develop ingrowths and higher levels of wall ingrowth deposition occur in abaxial- compared to adaxial-positioned PP TCs. Furthermore, the deposition of wall ingrowths was exclusively initiated on and preferentially covered the PP TC/SE interface, rather than the PP TC/CC interface, and only occurred in PP cells that were adjacent to SEs. Collectively, these results demonstrate the dominant impact of SEs on wall ingrowth deposition in PP TCs and suggest the existence of two sub-types of PP cells in leaf minor veins. Compared to PP cells, PP TCs showed more abundant accumulation of AtSWEET11-GFP, indicating functional differences in phloem loading between PP and PP TCs.

Mapping Pectic-Polysaccharide Epitopes in Cell Walls of Forage Chicory (Cichorium intybus) Leaves

The cell walls of forage chicory (Cichorium intybus) leaves are known to contain high proportions of pectic polysaccharides. However, little is known about the distribution of pectic polysaacharides among walls of different cell types/tissues and within walls. In this study, immunolabelling with four monoclonal antibodies was used to map the distribution of pectic polysaccharides in the cell walls of the laminae and midribs of these leaves. The antibodies JIM5 and JIM7 are specific for partially methyl-esterified homogalacturonans; LM5 and LM6 are specific for (1→4)-β-galactan and (1→5)-α-arabinan side chains, respectively, of rhamnogalacturonan I. All four antibodies labelled the walls of the epidermal cells with different intensities. JIM5 and JIM7, but not LM5 or LM6, labelled the middle lamella, tricellular junctions, and the corners of intercellular spaces of ground, xylem and phloem parenchyma. LM5, but not LM6, strongly labelled the walls of the few sclerenchyma fibres in the phloem of the midrib and lamina vascular bundles. The LM5 epitope was absent from some phloem parenchyma cells. LM6, but not LM5, strongly labelled the walls of the stomatal guard cells. The differential distribution of pectic epitopes among walls of different cell types and within walls may reflect the deposition and modification of these polysaccharides which are involved in cell wall properties and cell development.

Cellular and Metabolite Changes in the Secondary Phloem of Chinese Fir (Cuninghamia lanceolata (Lamb.) Hook.) during Dormancy Release

Wood in the cold temperate zone is the product of the alternation of the growing season and the dormant period of trees, but our knowledge of the process of dormancy release in trees remains limited. Chinese fir (Cuninghamia lanceolata (Lamb.) Hook.) was used to investigate cellular and metabolite changes in the secondary phloem tissue during dormancy release. The sampling dates were 2 March, 28 March, and 13 April. The microsections of wood-forming tissue were prepared using the paraffin embedding technique to observe the formation of cambium cells; metabolites in secondary phloem cells were extracted using a methanol/chloroform organic solvent system. The results showed that the secondary phloem consists of phloem fibers, sieve cells and phloem parenchyma. The cells were regularly arranged in continuous tangential bands and were in the order of Phloem fiber-Sieve cell-Phloem parenchyma-Sieve cell-Phloem parenchyma-Sieve cell-Phloem parenchyma-Sieve cell-Sieve cell-Phloem parenchyma-. The Chinese fir cambium was in dormancy on 2 March and 28 March, while on 13 April, it was already in the active stage and two layers of xylem cells with several layers of phloem cells were newly formed. The width of the cambium zone increased from 18.7 ± 5.7 μm to 76.5 ± 3.0 μm and the average radial diameter of sieve cells expanded from 15.4 ± 7.5 μm to 21.5 ± 7.4 μm after dormancy release. The cambium zone width and the average radial diameter of sieve cells before and after dormancy release were significantly different (p < 0.01). The phloem parenchyma cells without resin were squeezed and deformed by the sieve cells, and the width of the phloem during the active period was 197.0 ± 8.5 μm, which was larger than that during the dormant period. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)-based metabolomics was employed to analyze the secondary phloem of Chinese fir on 28 March and 13 April. Thirty-nine differential metabolites during dormancy release were detected. The results showed that the composition of Chinese fir metabolites was different before and after dormancy release. The relative increase in pyruvic acid and ascorbic acid contents proved that the rate of energy metabolism in Chinese fir increased substantially after dormancy release. Changes in cell development and the composition of metabolites revealed that the dormancy release of Chinese fir was at early April and the formation period of phloem tissue is earlier than xylem tissue.

Phloem loading via the abaxial bundle sheath cells in maize leaves

ABSTRACTLeaves are asymmetric, with differential functionalization of abaxial and adaxial tissues. The bundle sheath (BS) surrounding the vasculature of the C3 crop barley is dorsoventrally differentiated into three domains: adaxial structural, lateral S-type, and abaxial L-type. S-type cells seem to transfer assimilates towards the phloem. Here we used single-cell RNA sequencing to investigate BS differentiation in C4 maize. Abaxial BS (abBS) cells of rank-2 intermediate veins specifically expressed three SWEET sucrose uniporters (SWEET13a, b, and c) and UmamiT amino acid efflux transporters. SWEET13a, b, c were also identified in the phloem parenchyma (PP). Thus maize acquired a unique mechanism for phloem loading in which abBS cells provide the main pathway for apoplasmic sucrose transfer towards the phloem. This pathway predominates in veins responsible for phloem loading (rank-2 intermediate), while rank-1 intermediate and major veins export sucrose from the phloem parenchyma (PP) adjacent to the sieve element companion cell (SE/CC) complex, as in Arabidopsis. We surmise that abBS identity is subject to dorsoventral patterning and has components of PP identity. These observations provide first insights into the unique transport-specific properties of abBS cells and support for a modification to the canonical phloem loading pathway of maize, which may be generalizable to other C4 monocots.

Rescue and vegetative propagation of Eremanthus erythropappus (DC.) MacLeish in natural stand

The native stands of ‘candeia’ (Eremanthus erythropappus) have been explored through management plans due to the economic potential of essential oil. The rescue of adult trees, as well as the application of silvicultural techniques that favor the restoration of the stand, can contribute to the genetic conservation of this species. This study’s objective was to assess the efficiency of propagation techniques for the rescue of 26 matrices of ‘candeia’ in a natural managed stand and discussion about the rhizogenesis. In August 2017, trees were induced to regrowth by coppice, followed by exposure and scarification of roots. The emergence of shoots and morphology were evaluated according to the origin (i.e., stump or root). After that period, 19 matrices had their sprouts collected for the preparation of apical cuttings. Indole-3-butyric acid (IBA) was applied at the base of the cuttings. Cutting survival at greenhouse exit (GE), rooting at shade house exit (SHE), morphology and root anatomy were evaluated. In 189 days, the scarification of roots promoted 76.92% of budding. The percentage of sprouted matrices, number of shoots per matrice, length, diameter, and shoot length/diameter ratio increased over time. Only 12.2% of the cuttings survived in GE, and of these, 7.9% rooted in SHE. The cutting resulted in the formation of a clonal mini-garden of ‘candeia’, with seven of the 19 matrices submitted to propagation. The anatomical analyses showed that bud formation occurs from cell redifferentiation in the phloem parenchyma, and presence of crystals on the walls of the vessel elements of the secondary xylem. The shoots induction from scarification of roots could be used as a silvicultural practice for the reestablishment of the native fragments handle.

Unraveling the puzzle of phloem parenchyma transfer cell wall ingrowth

This article comments on: Wei X, Nguyen ST, Collings DA, McCurdy DW. 2020. Sucrose regulates wall ingrowth deposition in phloem parenchyma transfer cells in Arabidopsis via affecting phloem loading activity. Journal of Experimental Botany 71, 4690–4702.

Localization of (+)-Catechin in Picea abies Phloem: Responses to Wounding and Fungal Inoculation

To understand the positional and temporal defense mechanisms of coniferous tree bark at the tissue and cellular levels, the phloem topochemistry and structural properties were examined after artificially induced bark defense reactions. Wounding and fungal inoculation with Endoconidiophora polonica of spruce bark were carried out, and phloem tissues were frequently collected to follow the temporal and spatial progress of chemical and structural responses. The changes in (+)-catechin, (−)-epicatechin, stilbene glucoside, and resin acid distribution, and accumulation patterns within the phloem, were mapped using time-of-flight secondary ion mass spectrometry (cryo-ToF-SIMS), alongside detailed structural (LM, TEM, SEM) and quantitative chemical microanalyses of the tissues. Our results show that axial phloem parenchyma cells of Norway spruce contain (+)-catechins, the amount of which locally increases in response to fungal inoculation. The preformed, constitutive distribution and accumulation patterns of (+)-catechins closely follow those of stilbene glucosides. Phloem phenolics are not translocated but form a layered defense barrier with oleoresin compounds in response to pathogen attack. Our results suggest that axial phloem parenchyma cells are the primary location for (+)-catechin storage and synthesis in Norway spruce phloem. Chemical mapping of bark defensive metabolites by cryo-ToF-SIMS, in addition to structural and chemical microanalyses of the defense reactions, can provide novel information on the local amplitudes and localizations of chemical and structural defense mechanisms and pathogen–host interactions of trees.

Sucrose regulates wall ingrowth deposition in phloem parenchyma transfer cells in Arabidopsis via affecting phloem loading activity

In Arabidopsis thaliana, phloem parenchyma transfer cells (PPTCs) occur in leaf minor veins and play a pivotal role in phloem loading. Wall ingrowth formation in PPTCs is induced by the phloem loading activity of these cells, which is regulated by sucrose (Suc). The effects of endogenous versus exogenous Suc on wall ingrowth deposition, however, differ. Elevating endogenous Suc levels by increased light enhanced wall ingrowth formation, whereas lowering endogenous Suc levels by dark treatment or genetically in ch-1 resulted in lower levels of deposition. In contrast, exogenously applied Suc, or Suc derived from other organs, repressed wall ingrowth deposition. Analysis of pAtSUC2::GFP plants, used as a marker for phloem loading status, suggested that wall ingrowth formation is correlated with phloem loading activity. Gene expression analysis revealed that exogenous Suc down-regulated expression of AtSWEET11 and 12, whereas endogenous Suc up-regulated AtSWEET11 expression. Analysis of a TREHALOSE 6-PHOSPHATE (T6P) SYNTHASE overexpression line and the hexokinase (HXK)-null mutant, gin2-1, suggested that Suc signalling of wall ingrowth formation is independent of T6P and HXK. Collectively, these results are consistent with the conclusion that Suc regulates wall ingrowth formation via affecting Suc exporting activity in PPTCs.