The relationship between carotenoid accumulation and the expression of carotenoid biosynthetic genes during fruit maturation was investigated in three citrus varieties, Satsuma mandarin (Citrus unshiu Marc.), Valencia orange (Citrus sinensis Osbeck), and Lisbon lemon (Citrus limon Burm.f.). We cloned the cDNAs for phytoene synthase (CitPSY), phytoene desaturase (CitPDS), ζ-carotene (car) desaturase (CitZDS), carotenoid isomerase (CitCRTISO), lycopene β-cyclase (CitLCYb), β-ring hydroxylase (CitHYb), zeaxanthin (zea) epoxidase (CitZEP), and lycopene ϵ-cyclase (CitLCYe) from Satsuma mandarin, which shared high identities in nucleotide sequences with Valencia orange, Lisbon lemon, and other plant species. With the transition of peel color from green to orange, the change from β,ϵ-carotenoid (α-car and lutein) accumulation to β,β-carotenoid (β-car, β-cryptoxanthin, zea, and violaxanthin) accumulation was observed in the flavedos of Satsuma mandarin and Valencia orange, accompanying the disappearance of CitLCYe transcripts and the increase in CitLCYb transcripts. Even in green fruit, high levels of β,ϵ-carotenoids and CitLCYe transcripts were not observed in the juice sacs. As fruit maturation progressed in Satsuma mandarin and Valencia orange, a simultaneous increase in the expression of genes (CitPSY, CitPDS, CitZDS, CitLCYb, CitHYb, and CitZEP) led to massive β,β-xanthophyll (β-cryptoxanthin, zea, and violaxanthin) accumulation in both the flavedo and juice sacs. The gene expression of CitCRTISO was kept low or decreased in the flavedo during massive β,β-xanthophyll accumulation. In the flavedo of Lisbon lemon and Satsuma mandarin, massive accumulation of phytoene was observed with a decrease in the transcript level for CitPDS. Thus, the carotenoid accumulation during citrus fruit maturation was highly regulated by the coordination of the expression among carotenoid biosynthetic genes. In this paper, the mechanism leading to diversity in β,β-xanthophyll compositions between Satsuma mandarin and Valencia orange was also discussed on the basis of the substrate specificity of β-ring hydroxylase and the balance of expression between upstream synthesis genes (CitPSY, CitPDS, CitZDS, and CitLCYb) and downstream synthesis genes (CitHYb and CitZEP).

After several years in the juvenile phase, adult citrus trees show seasonal periodicity of flowering. A prolonged exposure to low temperature is one of the most important environmental cues for floral induction in citrus. In the present study, the expression of flowering-related genes during the annual cycle of flowering and inductive low-temperature treatment in Satsuma mandarin (Citrus unshiu Marc.) trees is investigated. Simultaneously, floral induction, which occurs before the period of morphological flower development, was estimated as the number of flowers after the forcing of sprouting by defoliation at 25 degrees C. The expression of citrus FLOWERING LOCUS T homologues, CiFT, showed a seasonal increase during the floral induction period and was also induced by an artificial low-temperature treatment (15 degrees C) at which floral induction occurred. By contrast, the mRNA level of CiFT did not show any distinct changes following a warm-temperature treatment (25 degrees C) for 2.5 months, during which time floral induction was completely suppressed. Changes in the expression of the citrus homologues of TERMINAL FLOWER 1, LEAFY, and APETALA1 did not show any correlation with floral induction in the field or under artificial low-temperature conditions. In juvenile seedlings of Satsuma mandarin, which does not flower even under inductive low-temperature conditions, the mRNA levels of CiFT were not affected by the low-temperature treatment, unlike adult tissues. These results suggest that low temperature promotes floral induction via the activation of CiFT transcription in adult Satsuma mandarin trees and that, in the juvenile plant, CiFT transcription does not respond to low temperature.