SummaryMonoclonal antibodies which recognize carbohydrate in arabinogalactan proteins (AGPs) have revealed that certain carbohydrate epitopes at the outer plasma membrane surface are developmentally regulated. Some epitopes are expressed according to cell position, and AGPs are thought to play a role in cell-cell interaction during development. This study demonstrates that sugar beet plasma membranes contain two subfamilies of AGPs, with apparent molecular masses of 82 and 97 kDa, and that each subfamily consists of a small number of acidic AGP isoforms. Excision of leaves generates three additional AGP complexes with apparent molecular masses of 120, 170 and 210 kDa, with the 170 kDa complex being the major form induced by excision. The addition of millimolar concentrations of H202 to a partially purified fraction of the 82 and 97 kDa AGPs also generates AGP complexes, with the 170 kDa complex as the major form. These results indicate that the plasma membrane AGPs are a target for endogenous H202.

A method for resolving plasma membrane associated arabinogalactan proteins (AGPs) has been developed. Plasma membranes purified by aqueous polymer two‐phase partitioning were first subjected to Triton X‐114 fractionation. The resulting water phase contained all detectable plasma membrane‐bound AGPs. Plasma membrane AGPs were then resolved in an SDS‐agarose gel electrophoresis system (SDS‐AGE). For separating plasma membrane AGP species of the same apparent molecular weight but with different net charge, a two‐dimensional electrophoresis system was used, utilizing isoelectric focusing in an immobilized pH gradient in the first dimension and SDS‐AGE in the second dimension. These methods enabled the separation of individual plasma membrane AGPs. In comparison, SDS‐PAGE methods left AGPs as unresolved high molecular‐weight smears. The methods described here may help to establish some basic features of AGPs, such as the number, organization, and protein and carbohydrate characteristics of plasma membrane AGPs, as well as the relationship between plasma membrane and extracellular AGPs.

In plants, cells differentiate according to their position with relation to their cell neighbours. Monoclonal antibody (MAb) probes to polysaccharide epitopes, present at the surfaces of all plant cells, have defined a family of proteoglycan antigens which signify cellular position. These MAbs have been used to sort the single cells present in carrot somatic cell cultures on the basis of the presence or absence of specific polysaccharide epitopes. This sorting allows embryo initial cells to be cultured among different cell collectives (based on their polysaccharide epitope expression) and thus in altered contextual backgrounds. These experiments have shown that specific populations of embryo initial precursor cells induce and sustain the early development of the embryo initials, revealing that the populations of different cell collectives which are defined by different polysaccharide epitopes (cell-context) serves important regulatory function in early plant development. Somatic embryo initials deprived of the influence of the cell collective-defined by the presence of the polysaccharide epitope recognised by the MAb JIM8-establish unorganised first divisions and develop as callus. However, in the presence of the JIM8-reactive cell collective, or medium conditioned by the collective, the initials develop into somatic embryos. This demonstrates that the cells defined by the JIM8 polysaccharide epitope are necessary to sustain the meristematic activity which drives the renewed development. Transfer of a cell-wall signal from the JIM8-reactive cells to cellular situations in carrot seedlings in which they would not normally occur (out-of-context signals) stimulates lateral root production, thus demonstrating that the inductive signal operative in suspension cultures can be reinterpreted by specific cells later in development and reinitiate meristematic activity. The communication between the precursor cells defined by JIM8 and embryo initials defines an early cell-cell interaction in developing carrot plants. Labelling of flower sections suggests that the same interaction exists between embryo apical and basal cells early in normal development.

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