三吉 一光

<p>Flower color patterns are attractive traits of floricultural plants. However, the mechanisms underlying these traits remain mostly unknown. Carnation (Dianthus caryophyllus L.) and interspecific hybrids thereof exhibit many flower color patterns involving white margins on reddish petals, as observed in the cultivar 'Minerva'. Flowers with white margins also have white abaxial surfaces. We studied the factors regulating the formation of white margins and a white abaxial surface in flowers through analyses of pigments and related gene expression. HPLC analysis revealed an absence of anthocyanins in white margins, although the accumulation of other flavonoid pigments (flavonols) was almost identical between the dark-red central and white marginal regions of petals. RNA-seq analyses of the dark red central regions and white marginal regions of 'Minerva' petals resulted in the extraction of 18 genes related to anthocyanin biosynthesis and transportation, including some transcription factors, as candidate regulatory genes for the formation of white margins. Further analysis of the expression of these genes by real-time RT-PCR and a comparison of two white-margin-flowered cultivars and three red-unicolor-flowered cultivars indicated that the expressions of two bHLH transcription factor genes and seven structural genes were positively correlated with anthocyanin accumulation. Although DcbHLH1, which is a homolog of JAF13 in Petunia ×hybrida, was expressed in both flower color groups, DcbHLH2, a homolog of AN1, was expressed only in white-marginal-flowered cultivars. Moreover, in the petals of 'Minerva', the expression of those nine genes was repressed in the abaxial epidermal layer without red pigmentation conferred by anthocyanins. Therefore, we could postulate that the localized repression of both bHLH genes is involved in the formation of white margins in carnation petals by inducing the absence of anthocyanin synthesis and that the white abaxial surface of the petals may result from similar regulatory mechanisms. In particular, DcbHLH2 could act as a key gene because of its restricted expression only in cultivars with white-margined flowers.</p>

We examined the effects of five antimitotic agents using Antirrhinum majus L. ‘Maryland True Pink’ on the induction of adventitious shoots resulted in increase of frequencies of chromosome doubling without plant growth regulators. Seeds were treated in vitro with 0, 16.5, 32.9, 65.8, 131.6, or 263.2 µM oryzalin (ORY), amiprofos-methyl (APM), butamifos (BUT), or propham (IPC) or 800, 1,600, 3,200, 6,400, or 12,800 µM colchicine (COL) for 7 day. ORY, COL and APM promoted induction of adventitious shoots on the hypocotyls at maximum frequencies of 57.6% with 16.5 µM ORY, 5.6% with 800 µM COL and 88.8% with 131.6 µM APM. ORY and COL also induced adventitious shoots on the epicotyls adjacent to the cotyledons, particularly at high concentrations, with a maximum frequency of 26.0% at 12,800 µM COL. APM treatment increased frequencies of tetraploids from 0.0 to 93.1%, with a positive correlation between the frequency and concentration. By contrast, ORY and COL induced tetraploids at frequencies of 16.0 to 54.6% and 4.0 to 59.4%, respectively, with peaks at both low and high concentrations of each. Correlation analysis revealed that frequencies of adventitious shoot formation could be useful as an index for the induction of tetraploids. These results showed that three of the antimitotic agents tested induced both adventitious shoot and tetraploid without plant growth regulators, indicating that antimitotic action may play a common role in the induction of adventitious shoot.

Chromosome numbers of wild species in the genus Epidendrum have previously been reported in the range of 2n=24-240. However, the ploidy levels of cultivars in the given genus originating from interspecific hybridization among several wild species have never been reported. To elucidate their ploidy levels, we analyzed the chromosome number of cultivars and related wild species. Four wild species showed a new record of chromosome number in the present study, namely, E. radicans 2n=38, 80, E. secundum 2n=60, and E. cinnabarinum 2n=64. Six cultivars examined were revealed to have more chromosomes than the wild species, with a range of 2n=84-164, suggesting that the cultivars have high polyploid levels. A significant correlation between the nuclear DNA amount and the chromosome number in wild species as well as cultivars was observed. Those cultivars originated from hybridization. Thus, the results from the present study suggest that the polyploid cultivars in this genus could have resulted from pollination involving unreduced gametes during breeding.

Dendrobium Stardust ‘Fire Bird’ is an interspecific hybrid between D. unicum and D. Ukon ‘Arai’ and is known to be sterile and prevents further breeding. Chromosome doubling is an essential method to increase the size of plant organs, such as flowers, and restore fertility of interspecific hybrids, which enables further breeding for ornamental traits. To induce chromosome doubling, the protocorm-like bodies (PLBs) of a given variety were treated in vitro for 6 to 48 h with 10 to 160 mg L−1 amiprofos-methyl (APM). Higher APM concentrations resulted in lower rates of survival of treated PLBs. Furthermore, subsequent growth of PLBs was hampered with prolonged APM treatment, although 10 and 20 mg L−1 APM was optimal for doubling chromosomes. Plantlets with a swollen and shorter stem were observed after the APM treatment and these plants were also chromosome-doubled and became a distinctive morphological marker for the primary selection of chromosome-doubled plants. Chromosome-doubled plants of D. Stardust ‘Fire Bird’ will be utilized for the future breeding of unique Dendrobium cultivars.