On the cover

Flower arrangements with blue chrysanthemums designed by H. Mogi (NARO). Blue chrysanthemums were created by introducing and co-expressing butterfly pea A3′5′GT and Canterbury bells F3′5′H. The blue coloration is due to intermolecular co-pigmentation between the newly synthesized anthocyanins and endogenous flavones (This issue, p. 79–87).

(N. Noda: NARO)

Carnation (Dianthus caryophyllus L.) is one of the main floricultural crops in Japan and worldwide. The vase life of cut ornamental flowers, including carnations, is important in determining their quality and consumers’ preference. To improve the vase life of carnation flowers, my group started a breeding research program in 1992 using conventional cross-breeding techniques. We repeatedly crossed and selected promising offspring with long vase life for seven generations, from 1992 to 2008. In 2005, we developed two cultivars, ‘Miracle Rouge’ and ‘Miracle Symphony’, with genetically determined long vase lives of 17.7 to 20.7 days (3.2 to 3.6 times that of ‘White Sim’) under standard conditions (23°C, 70% RH, 12-h photoperiod). Line 532-6 showed an ultra-long vase life averaging 27.8 to 32.7 days (4.6 to 5.4 times that of ‘White Sim’). We evaluated changes in ethylene sensitivity with flower senescence simply and accurately using a time-lapse video recorder. In 2010, we selected line 806-46b with both ultra-long vase life (27.1 days, 4.4 times that of ‘White Sim’) and ethylene resistance. Analyses using six cultivars and 123 selected lines from the 1st to the 7th generations revealed that the long vase life was strongly associated with a decrease in ethylene production.

Japanese gentians are the most important ornamental flowers in Iwate Prefecture and their breeding and cultivation have been actively conducted for half a century. With its cool climate and large hilly and mountainous area, more than 60% of gentian production in Japan occurs in Iwate Prefecture. Recent advances in gentian breeding and cultivation have facilitated the efficient breeding of new cultivars; disease control and improved cultivation conditions have led to the stable production of Japanese gentians. Molecular biology techniques have been developed and applied in gentian breeding, including the diagnosis of viral diseases and analysis of physiological disorders to improve gentian production. This review summarizes such recent approaches that will assist in the development of new cultivars and support cultivation. More recently, new plant breeding techniques, including several new biotechnological methods such as genome editing and viral vectors, have also been developed in gentian. We, therefore, present examples of their application to gentians and discuss their advantages in future studies of gentians.

Conventional breeding of cyclamen has relied on crossings among Cyclamen persicum cultivars without consideration of the scent of the flowers. Cyclamen purpurascens is a wild species with the most fragrant flowers in the genus Cyclamen. Allodiploid (2n = 2x = 41, AB) and allotriploid (2n = 3x = 65, AAB) plants have been produced from crosses of diploid and autotetraploid cultivars of C. persicum (2n = 2x = 48, AA; 4x = 96, AAAA) × diploid wild C. purpurascens (2n = 2x = 34, BB) by embryo rescue, but are sterile. Fertile allotetraploid (2n = 4x = 82, AABB) plants have been produced by chromosome doubling of the sterile allodiploids in vitro. Autotetraploid C. purpurascens (2n = 4x = 68, BBBB) has been produced by chromosome doubling of diploid C. purpurascens, and other fertile allotetraploids (2n = 4x = 82, AABB) have been produced from crosses of autotetraploid cultivars of C. persicum × autotetraploid C. purpurascens by embryo rescue. Commercial cultivars of fragrant cyclamen have been bred by conventional crosses among the allotetraploids. Mutation breeding using ion-beam irradiation combined with plant tissue culture has resulted in fragrant cyclamens with novel flower colors and pigments. In contrast, allotriploids (AAB) have not been commercialized because of seed sterility and poor ornamental value. The flower colors are determined by anthocyanins and flavonol glycosides or chalcone glucoside, and the fragrances are determined by monoterpenes, sesquiterpenes, phenylpropanoids, or aliphatics. Techniques for the production of fragrant cyclamen and knowledge of flower pigments and volatiles will allow innovation in conventional cyclamen breeding.

Lilies and tulips (Liliaceae family) are economically very important ornamental bulbous plants. Here, we summarize major breeding goals, the role of an integrated method of cut-style pollination and fertilization followed by embryo rescue and mitotic and meiotic polyploidization involved in new assortment development. Both crops have been subjected to extensive interspecific hybridization followed by selection. Additionally, spontaneous polyploidization has played a role in their evolution. In lilies, there is a tendency to replace diploids with polyploid cultivars, whereas in tulip a majority of the cultivars that exist today are still diploid except for triploid Darwin hybrid tulips. The introduction of molecular cytogenetic techniques such as genomic in situ hybridization (GISH) permitted the detailed studies of genome composition in lily and tulip interspecific hybrids and to follow the chromosome inheritance in interspecific crosses. In addition, this review presents the latest information on phylogenetic relationship in lily and tulip and recent developments in molecular mapping using different DNA molecular techniques.

Sweet pea (Lathyrus odoratus L.) is a major cut flower in Japan, generally grown in greenhouses in winter to spring. The wild-type sweet pea is a long-day summer-flowering plant. The day-neutral winter-flowering ability, which allows cut-flower production in Japan, is a recessive phenotype that emerged by spontaneous mutation. Although Japanese winter-flowering cultivars and additionally spring-flowering cultivars, which have semi-long-day flowering ability generated by crossing the winter- and summer-flowering cultivars, have superior phenotypes for cut flowers, they have limited variation in color and fragrance. In particular, variegated phenotypes do not appear in modern winter- and spring-flowering cultivars, only in summer-flowering cultivars. We try to expand the phenotypic diversity of Japanese cut flower cultivars. In the processes, we introduced the variegated phenotypes by crossing with summer-flowering cultivars, and succeeded in breeding some excellent cultivars. During breeding, we analyzed the segregation ratios and revealed the heredity of the phenotypes. Here we review the heredity of these variegated phenotypes and winter-flowering phenotypes and their related genes. We also describe how we introduced the trait into winter-flowering cultivars, tracing their pedigrees to show both phenotypes and genotypes of the progeny at each generation. This knowledge is useful for the efficient breeding of new variegated cultivars.

Genome information is useful for functional analysis of genes, comparative genomic analysis, breeding of new varieties by marker-assisted selection, and map-based gene isolation. Genome-related research in ornamentals plants has been relatively slow to develop because of their heterozygosity or polyploidy. Advances in analytical instruments, such as next-generation sequencers and information processing technologies have revolutionized biology, and have been applied in a large number and variety of species, including ornamental plants. Recently, high-quality whole genome sequences have been reported in plant genetics and physiology studies of model ornamentals, such as those in genus Petunia and Japanese morning glory (Ipomoea nil). In this review, whole genome sequencing and construction of high-density genetic linkage maps based on SNP markers of ornamentals will be discussed. The databases that store this information for ornamentals are also described.

Ornamental plants that have a rich variety of flower colors and shapes are highly prized in the commercial flower market, and therefore, mutant cultivars that produce different types of flowers while retaining their growth habits are in demand. Furthermore, mutation breeding is well suited for ornamental plants because many species can be easily vegetatively propagated, facilitating the production of spontaneous and induced mutants. The use of ion beams in mutation breeding has rapidly expanded since the 1990s in Japan, with the prospect that more ion beam-specific mutants will be generated. There are currently four irradiation facilities in Japan that provide ion beam irradiation for plant materials. The development of mutant cultivars using ion beams has been attempted on many ornamental plants thus far, and some species have been used to investigate the process of mutagenesis. In addition, progress is being made in clarifying the genetic mechanism for expressing important traits, which will probably result in the development of more efficient mutation breeding methods for ornamental plants. This review not only provides examples of successful mutation breeding results using ion beams, but it also describes research on mutagenesis and compares results of ion beam and gamma ray breeding using ornamental plants.

Flower color is the most important trait in the breeding of ornamental plants. In the floriculture industry, however, bluish colored flowers of desirable plants have proved difficult to breed. Many ornamental plants with a high production volume, such as rose and chrysanthemum, lack the key genes for producing the blue delphinidin pigment or do not have an intracellular environment suitable for developing blue color. Recently, it has become possible to incorporate a blue flower color trait through progress in molecular biological analysis of pigment biosynthesis genes and genetic engineering. For example, introduction of the F3′5′H gene encoding flavonoid 3′,5′-hydroxylase can produce delphinidin in various flowers such as roses and carnations, turning the flower color purple or violet. Furthermore, the world’s first blue chrysanthemum was recently produced by introducing the A3′5′GT gene encoding anthocyanin 3′,5′-O-glucosyltransferase, in addition to F3′5′H, into the host plant. The B-ring glucosylated delphinidin-based anthocyanin that is synthesized by the two transgenes develops blue coloration by co-pigmentation with colorless flavone glycosides naturally present in the ray floret of chrysanthemum. This review focuses on the biotechnological efforts to develop blue flowers, and describes future prospects for blue flower breeding and commercialization.

Transcription factors play important roles not only in the development of floral organs but also in the formation of floral characteristics in various plant species. Therefore, transcription factors are reasonable targets for modifying these floral traits and generating new flower cultivars. However, it has been difficult to control the functions of transcription factors because most plant genes, including those encoding transcription factors, exhibit redundancy. In particular, it has been difficult to understand the functions of these redundant genes by genetic analysis. Thus, a breakthrough silencing method called chimeric repressor gene silencing technology (CRES-T) was developed specifically for plant transcription factors. This method transforms transcriptional activators into dominant repressors, and the artificial chimeric repressors suppress the function of transcription factors regardless of their redundancy. Among these chimeric repressors, some were found to be inappropriate for expression throughout the plant body because they resulted in deformities. For these chimeric repressors, utilization of floral organ-specific promoters overcomes this problem by avoiding expression throughout the plant body. In contrast, attachment of viral activation domain VP16 to transcriptional repressors effectively alters into transcriptional activators. This review presents the importance of transcription factors for characterizing floral traits, describes techniques for controlling the functions of transcription factors.

Flower longevity is one of the most important traits for ornamental plants. Ethylene plays a crucial role in flower senescence in some plant species. In several species that show ethylene-dependent flower senescence, genetic modification targeting genes for ethylene biosynthesis or signaling has improved flower longevity. Although little is known about regulatory mechanisms of petal senescence in flowers that show ethylene-independent senescence, a recent study of Japanese morning glory revealed that a NAC transcription factor, EPHEMERAL1 (EPH1), is a key regulator in ethylene-independent petal senescence. EPH1 is induced in an age-dependent manner irrespective of ethylene signal, and suppression of EPH1 expression dramatically delays petal senescence. In ethylene-dependent petal senescence, comprehensive transcriptome analyses revealed the involvement of transcription factors, a basic helix-loop-helix protein and a homeodomain-leucine zipper protein, in the transcriptional regulation of the ethylene biosynthesis enzymes. This review summarizes molecular aspects of flower senescence and discusses strategies to improve flower longevity by molecular breeding.

Flowering time regulation has significant effects on the agricultural and horticultural industries. Plants respond to changing environments and produce appropriate floral inducers (florigens) or inhibitors (anti-florigens) that determine flowering time. Recent studies have demonstrated that members of two homologous proteins, FLOWERING LOCUS T (FT) and TERMINAL FLOWER 1 (TFL1), act as florigen and anti-florigen, respectively. Studies in diverse plant species have revealed universal but diverse roles of the FT/TFL1 gene family in many developmental processes. Recent studies in several crop species have revealed that modification of flowering responses, either due to mutations in the florigen/anti-florigen gene itself, or by modulation of the regulatory pathway, is crucial for crop domestication. The FT/TFL1 gene family could be an important potential breeding target in many crop species.

Chrysanthemum (Chrysanthemum morifolium Ramat.) is one of the most important floricultural crops in the world. Although the origin of modern chrysanthemum cultivars is uncertain, several species belonging to the family Asteraceae are considered to have been integrated during the long history of breeding. The flower color of ancestral species is limited to yellow, pink, and white, and is derived from carotenoids, anthocyanins, and the absence of both pigments, respectively. A wide range of flower colors, including purplish-red, orange, red, and dark red, has been developed by increasing the range of pigment content or the combination of both pigments. Recently, green-flowered cultivars containing chlorophylls in their ray petals have been produced, and have gained popularity. In addition, blue/violet flowers have been developed using a transgenic approach. Flower color is an important trait that influences the commercial value of chrysanthemum cultivars. Understanding the molecular mechanisms that regulate flower pigmentation may provide important implications for the rationale manipulation of flower color. This review describes the pigment composition, genetics, and molecular basis of ray petal color formation in chrysanthemum cultivars.

The Japanese morning glory (Ipomoea nil) and petunia (Petunia hybrida), locally called “Asagao” and “Tsukubane-asagao”, respectively, are popular garden plants. They have been utilized as model plants for studying the genetic basis of floricultural traits, especially anthocyanin pigmentation in flower petals. In their long history of genetic studies, many mutations affecting flower pigmentation have been characterized, and both structural and regulatory genes for the anthocyanin biosynthesis pathway have been identified. In this review, we will summarize recent advances in the understanding of flower pigmentation in the two species with respect to flower hue and color patterning. Regarding flower hue, we will describe a novel enhancer of flavonoid production that controls the intensity of flower pigmentation, new aspects related to a flavonoid glucosyltransferase that has been known for a long time, and the regulatory mechanisms of vacuolar pH being a key determinant of red and blue coloration. On color patterning, we describe particular flower patterns regulated by epigenetic and RNA-silencing mechanisms. As high-quality whole genome sequences of the Japanese morning glory and petunia wild parents (P. axillaris and P. inflata, respectively) were published in 2016, further study on flower pigmentation will be accelerated.

In a previous study, two genes responsible for white flower phenotypes in carnation were identified. These genes encoded enzymes involved in anthocyanin synthesis, namely, flavanone 3-hydroxylase (F3H) and dihydroflavonol 4-reductase (DFR), and showed reduced expression in the white flower phenotypes. Here, we identify another candidate gene for white phenotype in carnation flowers using an RNA-seq analysis followed by RT-PCR. This candidate gene encodes a transcriptional regulatory factor of the basic helix-loop-helix (bHLH) type. In the cultivar examined here, both F3H and DFR genes produced active enzyme proteins; however, expression of DFR and of genes for enzymes involved in the downstream anthocyanin synthetic pathway from DFR was repressed in the absence of bHLH expression. Occasionally, flowers of the white flowered cultivar used here have red speckles and stripes on the white petals. We found that expression of bHLH occurred in these red petal segments and induced expression of DFR and the following downstream enzymes. Our results indicate that a member of the bHLH superfamily is another gene involved in anthocyanin synthesis in addition to structural genes encoding enzymes.