Wachiraya Imsabai

The transcript abundance of an expansin gene in ripe sapodilla (Manilkara zapota) fruit is negatively regulated by ethylene

After harvest, mature fruit of sapodilla (Manilkara zapota van Royen) exhibit rapid softening. The decrease in fruit firmness was hastened by ethylene and delayed by 1-methylcyclopropene (1-MCP). Two genes encoding expansins (called MzEXP1 and MzEXP2) were isolated. In both cultivars studied (Makok-Yai and Kra-Suay), MzEXP1 was transiently expressed early during fruit development on the plant. This suggests that it is involved in cell wall loosening during early fruit growth. In cv. Makok-Yai, MzEXP2 was expressed between 1 day before harvest and day 4 after harvest. In cv. Kra-Suay, the expression of MzEXP2 started 8 weeks before the normal harvest stage, and ended on day 3 after harvest. When the fruit of both cultivars was treated with ethylene (50 µL L-1 for 20 h at 25°C) just after harvest, the expression of MzEXP2 became undetectable. After treatment with 1-MCP MzEXP2 mRNA was highly abundant until day 5 after harvest, when in controls the transcript abundance had become undetectable. The onset of MzEXP2 expression seems not regulated by ethylene, as the concomitant ethylene levels are very low. The data strongly indicate that the decrease of MzEXP2 transcript abundance is due to ethylene production by the fruit, which is by then high. The expression of MzEXP2 ceased, both in controls and in ethylene-treated material, when the fruit had reached a rather low threshold firmness. The data suggest that the protein has a supporting and cooperative role in fruit softening.

The effect of 1-methylcyclopropene (1-MCP) on expression of ethylene receptor genes in durian pulp during ripening

Rapid fruit ripening is a significant problem that limits the shelf life of durian, with ethylene having a major impact on the regulation of this event. Durian treated with ethephon ripened 3 d after treatment with increased pulp total soluble solids, ethylene production of the whole fruit and decreased pulp firmness compared to the control fruit. 1-MCP treatment delayed ripening by up to 9 d with inhibited accumulation of total soluble solids, color change, softening and ethylene production. Genes related to ethylene perception (DzETR1 and DzETR2) and the signaling pathway (DzCTR1, DzEIL1 and DzEIL2) in the pulp were investigated during this process, using qPCR to quantify changes in gene transcription. All candidate genes were significantly up-regulated in ripening durian pulp. Ethephon treatment increased the expression of DzETR1 and DzETR2 genes, while expression of DzCTR1, DzEIL1 and DzEIL2 were slightly affected. 1-MCP treatment significantly inhibited the expression of the DzETR2 and DzEIL1 genes. The promoters of DzETR2 genes were isolated and their activation by fruit transcription factors studied using transient expression in tobacco leaves. It was found that members of the kiwifruit and apple EIL1, EIL2 and EIL3 genes strongly activated the DzETR2 promoter. These results suggest that ethylene-induced ripening of durian is via the regulation of DzETR2 by EIL transcription factors.