Developmental complexity of arabinan polysaccharides and their processing in plant cell walls

Abstract

SUMMARYPlant cell walls are constructed from a diversity of polysaccharide components. Molecular probes directed to structural elements of these polymers are required to assay polysaccharide structures in situ, and to determine polymer roles in the context of cell wall biology. Here, we report on the isolation and the characterization of three rat monoclonal antibodies that are directed to 1,5-linked arabinans and related polymers. LM13, LM16 and LM17, together with LM6, constitute a set of antibodies that can detect differing aspects of arabinan structures within cell walls. Each of these antibodies binds strongly to isolated sugar beet arabinan samples in ELISAs. Competitive-inhibition ELISAs indicate the antibodies bind differentially to arabinans with the binding of LM6 and LM17 being effectively inhibited by short oligoarabinosides. LM13 binds preferentially to longer oligoarabinosides, and its binding is highly sensitive to arabinanase action, indicating the recognition of a longer linearized arabinan epitope. In contrast, the binding of LM16 to branched arabinan and to cell walls is increased by arabinofuranosidase action. The presence of all epitopes can be differentially modulated in vitro using glycoside hydrolase family 43 and family 51 arabinofuranosidases. In addition, the LM16 epitope is sensitive to the action of b-galactosidase. Immunofluorescence microscopy indicates that the antibodies can be used to detect epitopes in cell walls, and that the four antibodies reveal complex patterns of epitope occurrence that vary between organs and species, and relate both to the probable processing of arabinan structural elements and the differing mechanical properties of cell walls.

Escherichia coli Common Pilus (ECP) Targets Arabinosyl Residues in Plant Cell Walls to Mediate Adhesion to Fresh Produce Plants

“…To confirm the specificity, two arabinose-specific antibodies were used, LM6 and LM13. LM6 recognizes linear chains of (135)-␣-arabinan from pectin and some AGPs (39,43), whereas LM13 is specific to linear chains of (135)-␣-arabinan from pectin but importantly only to oligosaccharides longer than arabinotriose (38,44,45). As expected, LM6 recognized arabinose residues on the glycan arrays of all sizes, whereas LM13 recognized residues from arabinotriose to arabinoheptaose present on the array (Fig.…”

Pectin Biosynthesis Is Critical for Cell Wall Integrity and Immunity in Arabidopsis thaliana

“…This result confirmed that gae1 gae6 cell walls are reduced in HG. Using the same samples, we detected no difference from the wild type in binding of LM5, which recognizes a tetramer of (1-4)-b-Dgalactans found in RGI (Jones et al, 1997), LM6, which recognizes a linear pentasaccharide in (1-5)-a-L-arabinans found in RGI (Willats et al, 1998;Verhertbruggen et al, 2009b), LM8, which recognizes xylogalacturonan (Willats et al, 2004), or CCRC-M7, which recognizes an arabinosylated (1-6)-b-D-galactan epitope occurring on RGI (Puhlmann et al, 1994;Steffan et al, 1995;Pattathil et al, 2010) (Supplemental Figure 6). Collectively, these cell wall composition data show that gae1 gae6 plants are reduced in HG.…”

Loss of Cellulose Synthase-Like F6 Function Affects Mixed-Linkage Glucan Deposition, Cell Wall Mechanical Properties, and Defense Responses in Vegetative Tissues of Rice

“…Three-day-old coleoptiles, fully expanded leaves, and mature second stem internode pieces were fixed for 24 h at 4°C in 4% paraformaldehyde in 50 mM piperazine-N,N#-bis(2-ethanesulphonic acid), 5 mM EGTA, 5 mM MgSO4, pH 6.9 as described in Verhertbruggen et al (2009). The coleoptile pieces were cut from the basal region of the organ and were 4.0-and 6.0-mm long for longitudinal and transverse sections, respectively.…”