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Phloem fibres in  Mallotus japonicus  Müll. Arg. were found to have a multi-layered structure that is S 1  + S 2  +  n (G + L), where a non-lignified gelatinous layer (G) and a lignified layer (L) are formed alternately and  n  indicates the number of repetitions of these two layers. The aim of this study was to determine the process of xylan deposition and lignification in the multi-layered cell walls of phloem fibres. The formation process of the multi-layered structure of secondary phloem fibres was examined by light microscopy, ultraviolet microscopy and transmission electron microscopy. The distribution of glucuronoxylan was examined by immunoelectron microscopy. The activity of peroxidase was also determined using metal-enhanced diaminobenzidine substrates. Immunolabelling of glucuronoxylan occurred in lignified cell wall layers, except in the compound middle lamella (CML), i.e., the S 1 , S 2  and L layers but not the G layers. Change in immunolabelling density suggests that xylan deposition in these lignified layers occurs appositionally, i.e., xylan is deposited into the lignified layers directly and not by a penetrative mechanism, and deposition does not occur after the layers are fully deposited. Peroxidase activity was found in CML including cell corners during S 2  layer formation, then in developing G layers during G layer formation. Peroxidase activity was also found in the thin L layers that formed recently and was not found in the L layers already present. Xylan labelling was not found in the thin L layers that formed recently but did occur in L layers that developed earlier. Lignification of the S 1  and S 2  layers continued during the formation of the G layers, whereas in the L layers it finished just after deposition of the L layer.

Xylan deposition and lignification in the multi-layered cell walls of phloem fibres in Mallotus japonicus (Euphorbiaceae)