Toves, Peter J.
Amore, Teresita D.
Tropical Plant and Soil Sciences
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Anthurium is the most important cut flower in the Hawaii floriculture industry. Conventional breeding for desirable vase life, shape, and color has been the mainstay for keeping growers in Hawaii competitive in the global market. No blue anthuriums exist and cannot be made via conventional breeding since anthuriums lack the genes to produce delphinidin-derived anthocyanins. The objectives of this research were to expand our understanding of the biochemical and histological aspects of flower color that are essential for developing strategies for breeding, conduct transient gene expression in anthurium spathes, and to explore genetic engineering of the anthurium flavonoid pathway for novel color development. Flower color is influenced by pH, since anthocyanins exist in different forms depending on the pH of the vacuole. A survey of spathe pH in 50 hybrid selections and cultivars revealed that green spathes had the highest average pH (6.14), followed by whites (5.96), purples (5.82), pinks (5.78), corals (5.72), reds (5.46), and oranges (5.40). Flower color is influenced by the distribution and combinations of anthocyanins in the floral tissue. Anthocyanins were distributed in the hypodermis and mesophyll in red, pink, orange and coral anthurium spathes, with some hybrid selections having very few pigmented cells in the epidermal layer. The spatial distribution of anthocyanins was more expansive in purple anthurium spathes, and was observed in the adaxial and abaxial epidermis, and in the adaxial and abaxial hypodermis of some selections/cultivars. White and green spathes lacked anthocyanins; however green spathes contained substantial amounts of chlorophyll. The pH and spatial distribution data can help to make informed decisions when using the surveyed anthurium cultivars/selections for breeding, since pH can affect color and spatial distribution of anthocyanins can affect the perceived color intensity. Since the development of stable transformants with genes for novel flower color is a lengthy process with low transformation efficiency, we tested agroinfiltration-mediated transient expression to assess functionality of the structural gene F3’5’H and the transcription factors Delila and Rosea1 in 18 selections/cultivars. The effects of detached and attached spathes, older and younger spathes, the effect of full spectrum and enhanced LED, and sonication were tested in addition to agroinfiltration with the color constructs. The transient transformation results obtained were inconsistent and not conclusive for positive transient expression, since some spathes infiltrated without the color gene or transcription factors developed blue coloration despite the lack of F3’5’H or transcription factors Delila and Rosea1 . In this study, calli of seven hybrid selections or cultivars were co-cultivated with Agrobacterium tumefaciens AGL0 with one of the following plasmids with the NPTII selection gene: 1) pJAM1983 with the gene F3'5'H from petunia, 2) pJAM1889 with the gene Delila from Antirrhinum majus, 3) pJAM1463 harboring the gene Roseal from A. majus. Of the 372 samples tested via Polymerase Chain Reaction (PCR) (some made by Hawaii Agriculture Research Center (HARC), others made in this research), eight ‘Marian Seefurth’ samples from HARC tested positive for the NPTII antibiotic resistance gene and six of those samples tested positive for F3’5’H.
Horticulture, anthocyanin, anthurium, genetic engineering, histology
178 pages
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