Chemical studies on different color development in blue- and red-colored sepal cells of Hydrangea macrophylla.

To clarify the cause of the difference in blue and red color development of hydrangea sepals, Hydrangea macrophylla, we analyzed the organic and inorganic components in the colored cells. To obtain colored protoplasts, each blue and red sepal tissue was treated with a combination of cellulase and pectinase, and then from the suspension of the olored and colorless protoplast mixture colored cells of the same hue were collected with a micro-pipette. The content of organic components (delphinidin 3-glucoside, chlorogenic acid, neochlorogenic acid and 5-O-p-coumaroylquinic acid) and Al(3+) in each colored cell was quantified respectively by semimicro-HPLC and graphite furnace atomic absorption spectroscopy (GFAAS). In the blue cells 13 eq. of 5-O-acylquinic acids and 1.2 eq. of Al(3+) to anthocyanin were contained. Contrary to this result, in the red cells, only 3.6 eq. of 5-O-acylquinic acids and 0.03 eq. of Al(3+) were detected. A reproduction experiment of each blue and red sepal color by mixing those components concluded that, for blue coloration, both 5-O-acylquinic acids and Al(3+) were essential.

Identification of aluminum species in an aluminum-accumulating plant, hydrangea (Hydrangea macrophylla), by electrospray ionization mass spectrometry.

The use of electrospray ionization mass spectrometry (ESI-MS) in negative ion mode was investigated as a direct probe for identifying Al species in Al-accumulating hydrangea (Hydrangea macrophylla) samples. Cell sap solutions of hydrangea leaves were purified using Sephadex G-10 liquid chromatography and each fraction was analyzed using ESI-MS and ESI-MS/MS to identify Al species. In hydrangea leaves, a 1:1 Al-citrate complex was found as [AlH(-1)cit](-) (m/z 215), where H(3)cit denotes citric acid. This result is consistent with that of Ma et al. who used (27)Al-NMR.