Pigment Composition Comprising Anthocyanic Vacuolar Inclusions
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example 1
Isolation of AVIs from Plants
Plant Material
[0118]The lisianthus lines used in this study were lines #54 and Wakamurasaki (see Deroles et al. 1998 for further details). Fully open flowers were harvested and washed with distilled water to remove potential contaminants such as pollen grains. Various petal regions were excised with a razor blade for further study.
[0119]Lisianthus AVIs are found as one or more large irregular shapes in the vacuoles of adaxial epidermal cells in the throat of the flower. The lisianthus AVIs could be removed from cell vacuoles using narrow glass micropipettes. They remained intact, and adhered firmly to glass indicating that they are structural rather than localized concentrations of dissolved anthocyanins. In addition, electron microscopy images show that the AVI body is not membrane bound.
Protoplast Generation and Isolation
[0120]Protoplasts were generated from macerated inner petal material as described by Morgan (1998). Material from 10 flowers was adde...
example 2
Stability of AVI-Bound Anthocyanins in a Range of pH Environments
[0123]AVIs isolated from lisianthus were resuspended in a McIlvaines buffer (Citric Acid / Sodium Phosphate) array from pH 3.0 to 8.0. The samples were left at room temperature and lighting conditions (fluorescent / natural light) for up to 7 days to determine the stability and colourfastness of the AVI-bound anthocyanins. A spectrum of each sample (350-750 nm) was taken at daily intervals to determine the hue and intensity of the AVI-bound anthocyanins over time. In the control experiment, anthocyanins were extracted from the AVIs and subjected to the same pH conditions.
[0124]As shown in FIG. 1 the anthocyanins in this experiment have maintained their colour over a wide pH range and for a significant period of time. All the other treatments do not change significantly until 7 days where the samples at more alkaline pH levels (pH 6-8) begin to fade.
[0125]Anthocyanins were then extracted from the lisianthus AVIs and resuspe...
example 3
Stability of AVI-Bound Anthocyanins in Commercial Natural Yoghurt
[0126]AVIs from lisianthus were mixed with a commercial natural yoghurt sample (pH 4.18) obtained from a supermarket in order to determine their ability to maintain colour in a processed food environment. The samples were left at room temperature under room lighting conditions for 24 to 48 hours.
[0127]As shown in FIG. 3, the lisianthus AVIs were able to maintain their colour over a 48-hour period.
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