HORTSCIENCE 45(12):1768–1772. 2010. in roses through the introduction of the flavo- noid 3#,5#-hydroxylase (F3#5#H) gene were unsuccessful. Katsumoto et al. (2007) generated Spathe Color Variation in Anthurium blue roses by placing the F3#5#Hgeneinto a genetic background with higher vacuolar pH andraeanum Hort. and Its Relationship and high flavonol content. Griesbach (2005) observed that although flavonols and an to Vacuolar pH appropriate pH are important in obtaining blue orchids, the more important of the two Omaira Avila-Rostant, Adrian M. Lennon, factors was vacuolar pH. Creation of blue and Pathmanathan Umaharan1 orchids, he suggested, would therefore require Department of Life Sciences, Faculty of Science and Agriculture, The University the screening of germplasm for high floral pH and combining the independent codominantly of the West Indies, St. Augustine Campus, College Road, Republic of Trinidad inherited high pH genes into a single genotype and Tobago (Griesbach, 2005). Additional index words. anthocyanin, chromospectrometry, flavonoids, flower color, phenyl Anthurium andraeanum Linden ex Andre´ is a tropical ornamental species with a limited propanoid pathway range of spathe colors based on anthocyanin Abstract. The relationship between vacuolar pH in Anthurium andraeanum (Hort.) and pigments that vary from orange to red. The spathe color, cultivar, developmental stage, spathe location, spathe surface differences, color range has been expanded through in- and time after harvest was investigated with the overall long-term objective of developing terspecific hybridization with species belong- a methodology for engineering blue-colored spathes. Chromospectral analysis of the ing to the section Calomystrium (Kamemoto pigmentation was also studied. Six experiments were conducted involving 23 cultivars and Kuehnle, 1996) through the creation of a of anthurium with each experiment arranged in a randomized complete block design with species complex referred to as A. andraeanum five replications. Spathe color was associated with vacuolar pH with the whites and greens (Hort.). having the highest pH (average 5.65) followed by corals (5.38), pinks (5.20), reds (5.10), and Iwata et al. (1979) reported that the major oranges (4.5). In general, there was correspondence between the lightness of the pigmen- spathe colors, red to pink and orange to coral, tation (L*) and the pH values with the lighter colors having higher pH values. There were, are determined by two anthocyanins, pelargo- however, significant differences among cultivars within the color groups. Whereas spathe nidin 3-rutinoside (pelargonidin 3-rhamonsyl- pH decreased with aging, there was no difference in the spectral data, suggesting that factors glucoside) and cyanidin 3-rutinoside (cyanidin other than anthocyanin content may be contributing to the pH difference. There were no 3-rhamnosyl glucoside) found exclusively in differences in pH between locations sampled on the spathe nor between the spathe surfaces the hypodermal layers of abaxial and adaxial provided there were no differences in color intensity (L*). The pH increased with vase life surfaces of the spathe (Ehrenberger and in two of three cultivars tested with pH values showing an association with increases in a* Kuehnle, 2003; Higaki et al., 1984). Whereas and b* (chromospectral data) reflecting a bluing effect. The importance of the results to pelargonidin 3-rutinoside is responsible for engineering blue-colored spathes in anthurium is discussed. orange and coral spathes, both pelargonidin 3-rutinoside and cyanidin 3-rutinoside are found in red and pink spathes. Coral and pink Flower color in plants is determined by lated with pigment composition. For instance, spathes have lower concentrations of antho- pigments such as aurones, anthocyanins, and in rose cultivars that contained cyanidin, flowers cyanins in comparison with orange and red carotenoids (Davis and Schwinn, 1997; Schijlen varied from red to lavender and those with counterparts (Iwata et al., 1985). The white et al., 2004). Anthocyanins, the major pigment peonidin varied from red to purple (Griesbach, spathes lack both anthocyanins but contain in Anthurium andraeanum (Hort), are the prod- 1996). Similar observations were made in tulip colorless flavone C-glycosides (Williams et al., uct of flavonoid biosynthesis and are divided (Nieuwhof et al., 1989) and hydrangea (Yoshida 1981). Conspicuous lack of delphinidins or into cyanidins and their derivatives that produce et al., 2003). peonidin (a cyanidin derivative) account for colors ranging from red to purple (Griesbach, The role of pH on flower color has been the lack of mauves, purples, and blues (Iwata 1996); pelargonidins and their derivatives that well established in many plant species, includ- et al., 1985). produce colors ranging from coral to orange ing hydrangea, petunia, morning glory, orchids, Recently the genetics and biochemistry of (Iwata et al., 1979); and delphinidins and their and rose. Although the sepals of hydrangea the anthurium flavonoid biosynthetic pathway derivatives that produce colors from blue to have only one anthocyanin, delphinidin- have been characterized with the intention of deep red (Asen and Siegelman, 1957). 3-glucoside, the color displayed varies from creating colors in the blue range (Collette Researchers have stated that the ultimate red to blue (Asen and Siegelman, 1957) with et al., 2004; Elibox and Umaharan, 2008a). color displayed is dependent not only on the corresponding changes in vacuolar pH from However, no studies exist with regard to var- pigment present, but also on a number of factors 3.3 to 4.1 (Yoshida et al., 2003). In petunia, iation in pH in anthurium cultivars. With the including cell shape (Noda et al., 1994), pres- the inheritance of flower color was attributed objective of identifying suitable candidates ence of various metal ions (Shoji et al., 2007), to the combined effect of anthocyanin pig- for transformation toward generating blues in stacking of anthocyanins with copigments such mentation and pH, the latter being controlled anthurium, this study investigates the relation- as flavones and flavonols (Goto and Kondo, by two independent codominant genes, Ph1 ship between epidermal vacuolar pH and a 1991), and pH (Harborne, 1988; Katsumoto and Ph2 (Griesbach, 1996). In morning glory, number of plant factors, including cultivar, et al., 2007; Stewart et al., 1975). As a result, flower color varied from reddish purple buds spathe color, developmental stage of the spathe, in some species, the flower color is not corre- to blue flowers with an increase in vacuolar location of anthocyanin within the spathe, dif- pH from 6.6 to 7.7, a change believed to be ferences between the abaxial and adaxial sur- driven by a Na+(K+)/H+ exchanger (Yamaguchi faces, and postharvest changes. Received for publication 24 June 2010. Accepted et al., 2001; Yoshida et al., 2005). In Phalae- for publication 23 Sept. 2010. nopsis pulcherrima, blue-flowered cultivars Materials and Methods This study was funded by a research grant from the had a pH more alkaline (pH 5.7) compared Government of Trinidad and Tobago. We thank the management and staff of Kairi with the purple form of the species (pH 4.9) Plant material. Spathes were collected Blooms Farm for providing the material for the with high pH being governed by a single from 23 cultivars of Anthurium andraeanum research. recessive gene (Griesbach, 1997). (Hort.) maintained at Kairi Blooms Ltd., a 1To whom reprint requests should be addressed; Although roses exhibit a variety of colors, commercial anthurium farm situated in Car- e-mail [email protected]. they lack blues. Early attempts to generate blues apo Village, Arima, Trinidad. The collected 1768 HORTSCIENCE VOL. 45(12) DECEMBER 2010 | BREEDING,CULTIVARS,ROOTSTOCKS, AND GERMPLASM RESOURCES samples were placed in a cooler and trans- of tissue maturity on pH. The experiment was Pearson’s product moment correlation coeffi- ported to the laboratory. All blooms were replicated five times. cient or regression analysis (NCSS, 2001). harvested between 0700 to 0800 HR during the In the fourth experiment, epidermal peels months of January to February. Cultivars Pier- from three discs each from the abaxial and Results rot, Tropical, and Success were harvested at adaxial surfaces were investigated with five three developmental Stages 6-1 (newly opened), replications in two cultivars (Tropical, So- There were significant (P < 0.05) differ- 6-2 (spadix ½ mature), and 6-3 (spadix 3/4 nata) as described. ences in pH between the various color groups mature), as defined by Collette (2002). Cultivar In the fifth experiment, three cultivars (Table 1) with the highest pH recorded for Tropical was harvested at five stages (Stages (President, Senator, and KAIRI3674) with green-spathed cultivars followed by white, 2to6)todeterminetheeffectofstageoncolor ‘‘obake’’ (bicolored) spathes were used. These coral, pinks/reds, and orange, in that order. formation. cultivars produce bicolored spathes with vari- There was no significant difference (P > 0.05) pH measurement. A random sample of ous anthocyanins at the center of the spathe between red- and pink-spathed cultivar groups. five discs from 10 discs (50 mm2) obtained and no anthocyanins at the periphery. ‘Pres- Cultivar differences in vacuolar pH within the from each spathe using a cork borer was used ident’ and ‘KAIRI3674’ have a dark pink various colors were significant (P < 0.05) for as an experimental unit in the studies. The center, whereas ‘Senator’