Histolocalisation of the oil and pigments in the pumpkin seed

Pumpkin seed oil has a distinct taste and a characteristic green‐red colour. A green layer (chlorenchyma) can be observed in the cross section of the seed on the inner side of the testa. This layer is considered to be a source of protochlorophyll pigment that gives colour to the pumpkin seed oil. However, the histolocalisation of the pigment by selective method was not yet studied. In this study, we localised the oil to the small lipid droplets in the cotyledon cells by Rhodamine 6G lipophilic stain. Furthermore, we localised protochlorophyll to chlorenchyma by microscopic fluorimetry. Emission maximum of protochlorophyll was 700 nm when measured in the seed, but it shifted to 630 and 655 nm (two peaks) when measured in pumpkin seed oil or in the extract.