Succinate dehydrogenase activity of external and internal hyphae of a vesicular-arbuscular mycorrhizal fungus,Glomus mosseae (Nicol. & Gerd.) Gerdmann and Trappe,during mycorrhizal colonization of roots of leek (Allium porrum L.), as revealed by in situ histochemical staining

The succinate dehydrogenase (SDH) activity of hyphae of the vesicular-arbuscular (VA) mycorrhizal fungus Glomus mosseae (Nicol. & Gerd.) Gerdmann and Trappe, in symbiotic association with leek (Allium porrum L.) roots, was investigated by histochemical staining in situ. Leek seedlings were transplanted to sand culture and inoculated with spores of G. mosseae placed just below the base of the stem. At intervals (14, 25, 35 and 60 days) after transplanting, the growth medium of seedlings was flooded with nitro blue tetrazolium chloride solution, thereby displacing the nutrient solution. This allowed sites of SDH activity of external and internal fungal structures of the mycorrhizas to be stained without physically disturbing the symbiotic system. After counterstaining harvested roots and mycelium with acid fuchsin, it was possible to differentiate clearly metabolically active and inactive regions of the fungus. The lengths of external hyphae and infected root both increased nearly exponentially, and were in constant proportion (1.4 m hyphae per cm of infected root) for up to 60 days. The percentage length of external hyphae with SDH activity remained almost constant (80%). In each infected length of root there was a gradation of SDH activity from inactive distal (older) hyphae to uniformly active proximal (younger) hyphae. These findings are discussed in relation to the symbiotic activity of the mycobiont.Deceased     

Effect of anti-wall protein antibodies on auxin-induced elongation, cell wall loosening, and β-D-glucan degradation in maize coleoptile segments

Antiserum raised against the LiCl extract of maize shoot cell walls suppresses auxin-induced elongation of maize coleoptile segments. A series of polyclonal antibodies were raised against protein fractions separated from the LiCl extract of maize ( Zea mays L. cv. B73 x Mo17) coleoptiles by SP-Sephadex and Bio-Gel P-150 chromatography. To understand the role of cell wall proteins in growth regulation, the effect of these antibodies on auxin-induced elongation and changes in the cell walls of maize coleoptiles was examined. Four of the fractions prepared reacted with the antiserum raised against the total LiCl extract and effectively suppressed its growth-inhibiting activity. Only these fractions contained the proteins responsible for eliciting growthinhibiting antibodies. The antibodies capable of growth inhibition of auxin-induced elongation of segments also inhibited auxin-induced cell wall loosening (decrease in the minimum stress-relaxation time of the cell walls) of segments. The antibodies raised against one of the protein fractions separated by SP-Sephadex inhibited the autolytic reactions of isolated cell walls and the auxin-induced decrease in (1→3), (1→4)-β-D-glucans in the cell walls. Thus, the degradation of β-D-glucans by cell wall enzymes may be associated with the cell wall loosening that is responsible for cell elongation. Because the other antibodies did not influence the auxin-induced degradation of (1→3), (1→4)-β-D-glucanses, β-D-glucanases and other cell wall enzymes may cooperate in regulation of cell elongation in maize coleoptiles.     

Antibodies as probes for the study of location and metabolism of (1→3), (1→4)-β-D-glucans

Polyclonal antibodies, raised against ((1→3), (1→4)-β-D-glucans from oat ( Avena sativa L.) caryopsis, were used to investigate the location and the metabolism of mixed-linked β-D-glucans. The binding of these antibodies to the cell walls of oat coleoptiles was shown by an indirect fluorescence method. Distinct fluorescent regions were observed along the inner layers of the walls of each cell. The preimmune serum or antibodies pretreated with oat caryopsis β-D-glucans did not react with the cell walls. Glucan antibodies were bound to the walls of other Poaceae coleoptiles as well as to those from oat mesocotyls and roots, whereas they were not bound to the walls of some dicotyledons tested. The relative glucan content of the cell walls of oat coleoptiles as determined by β-D-glucanase (EC 3.2.1.73) treatment was maximum between day 3 and 4 after soaking, but it declined during further elongation. A rapid decrease in glucan content was observed in excised coleoptiles when auxin or β-D-glucanase was present. There was a clear correlation between the glucan content expressed on a basis of cell wall polysaccharides and the amount of the antibodies bound to the cell walls. These results indicate that the antibodies are useful probes to detect and determine (1→3), (1→4)-β-D-glucans of cell walls.