cDNA clones encoding homologues of expansins, a class of cell wall proteins involved in cell wall modification, were isolated from various stages of growing and ripening fruit of tomato (Lycopersicon esculentum). cDNAs derived from five unique expansin genes were obtained, termed tomato Exp3 to Exp7, in addition to the previously described ripening-specific tomato Exp1 (Rose et al. (1997) Proc Natl Acad Sci USA 94: 5955-5960). Deduced amino acid sequences of tomato Exp1, Exp4 and Exp6 were highly related, whereas Exp3, Exp5 and Exp7 were more divergent. Each of the five expansin genes showed a different and characteristic pattern of mRNA expression. mRNA of Exp3 was present throughout fruit growth and ripening, with highest accumulation in green expanding and maturing fruit, and lower, declining levels during ripening. Exp4 mRNA was present only in green expanding fruit, whereas Exp5 mRNA was present in expanding fruit but had highest levels in full-size maturing green fruit and declined during the early stages of ripening. mRNAs from each of these genes were also detected in leaves, stems and flowers but not in roots. Exp6 and Exp7 mRNAs were present at much lower levels than mRNAs of the other expansin genes, and were detected only in expanding or mature green fruit. The results indicate the presence of a large and complex expansin gene family in tomato, and suggest that while the expression of several expansin genes may contribute to green fruit development, only Exp1 mRNA is present at high levels during fruit ripening.

Expansins are extracellular proteins that facilitate cell wall extension, possibly by disrupting hydrogen bonding between hemicellulosic wall components and cellulose microfibrils. In addition, some expansins are expressed in non-growing tissues such as ripening fruits, where they may contribute to cell wall disassembly associated with tissue softening. We have identified at least three expansin genes that are expressed in tomato (Lycopersicon esculentum Mill.) seeds during germination. Among these, LeEXP4 mRNA is specifically localized to the micropylar endosperm cap region, suggesting that the protein might contribute to tissue weakening that is required for radicle emergence. In gibberellin (GA)-deficient (gib-1) mutant seeds, which germinate only in the presence of exogenous GA, GA induces the expression of LeEXP4 within 12 hours of imbibition. When gib-1 seeds were imbibed in GA solution combined with 100 microM abscisic acid, the expression of LeEXP4 was not reduced, although radicle emergence was inhibited. In wild-type seeds, LeEXP4 mRNA accumulation was blocked by far-red light and decreased by low water potential but was not affected by abscisic acid. The presence of LeEXP4 mRNA during seed germination parallels endosperm cap weakening determined by puncture force analysis. We hypothesize that LeEXP4 is involved in the regulation of seed germination by contributing to cell wall disassembly associated with endosperm cap weakening.

Cyst nematodes induce specific syncytial feeding structures within the root which develop from an initial cell by successive incorporation of neighbouring cells through local cell wall dissolutions followed by hypertrophy of included cells. Expansins are known to induce cell wall relaxation and extension in acidic pH, and they are involved in many processes requiring wall modification from cell expansion to cell wall disassembly. We studied the expression pattern of tomato (Lycopersicon esculentum L., cv. Money Maker) expansins during development of syncytia induced by the potato cyst nematode (Globodera rostochiensis Woll.). Based on semi-quantitative reverse transcription-polymerase chain reaction, two expansin genes, LeEXPA4 and LeEXPA5, were selected for detailed examinations because their expression was either elevated in infected roots (LeEXPA4) or specifically induced in the root upon nematode infection (LeEXPA5). Both genes have distinct spatial and temporal expression patterns that may reflect their different roles in syncytium development. LeEXPA4 transcripts were localized predominantly in parenchymatous vascular cylinder cells surrounding syncytia. This finding suggests that LeEXPA4 might be involved in cell wall disassembly or relaxation, mediating syncytium expansion and/or development of conductive tissues. By contrast, LeEXPA5 transcripts were localized in enlarging syncytial elements. Similarly, in immunogold localization experiments, polyclonal antibodies localized the LeEXPA5 protein in cell walls of syncytial elements. This expression pattern suggests that LeEXPA5 gene is specifically involved in enlargement of cells incorporated into syncytium.