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Originated from Argyranthemum Frutescens Parentages

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Originated from Argyranthemum Frutescens Parentages Chromosome Botany (2014) 9: 97-112 © Copyright 2014 by the International Society of Chromosome Botany Identifying, discriminating and isolating cultivars of ‘Marguerites’ originated from Argyranthemum frutescens parentages and their intergeneric and interspecific hybridities by DNA markers amplified by RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter-Simple Sequence Repeat)* Masahiro Morikawa1,4, Takahiro Ｍuto2, Arnoldo Santos-Guerrra3 and Katsuhiko Kondo1,5 1Laboratory of Plant Genetics and Breeding Science, Department of Agriculture, Faculty of Agriculture, Tokyo University of Agriculture, 1737 Funako, Atsugi City, Kanagawa Prefecture 243-0034, Japan; 2Plant Cultivation and Breeding Section, Izu Agricultural Research Center, Shizuoka Prefectural Reserch Institute of Agriculture and Forestry, Izu Peninsula 3012 Inatori, Izuchou, Kamo-Gun, Shizuoka Prefecture 413-0411, Japan; 3Head of Botany Department, Orotava Botanical Garden, Tenerife, Canary Island, Spain; 4The Present Address: Saitama Genshu Ikuseikai Co., Ltd., 2616 Niibori, Kuki City, Saitama Prefecture 346-0105, Japan 5Author for Correspondence: [email protected] Received May 5, 2014; accepted September 30, 2014 ABSTRACT. Abilities of the polymorphic DNA markers amplified by RAPD (Randomly Amplified Polymorphic DNA) and ISSR (Inter Simple Sequence Repeat) were studied, detected and systematized breeding lines and expanding molecular variabilities and molecular phylogenetic lines of the cultivarietal ‘Marguerites’ of the Argyranthemum lineage and especially the intergeneric hybrids of ‘Marguerite’ X Glebionis carinata Tzvelev or G. coronaria (L.) Spach. The reproducible polymorphic bands were generated by RAPD-PCR using RAPD primer OPC-04, in respective species of Glebionis and the intergeneric hybrids of the ‘Marguerites.’ The primer OPC-04 made it possible to identify either two intergeneric hybrids of ‘Marguerite’ X G. carinata or ‘Marguerite’ X G. coronaria. The polymorphic bands were amplified by ISSR-PCR in the breeding lines and the cultivars of the ‘Marguerite,’ while they were not amplified by RAPD. A specific band was confirmed by ISSR-5 primer in the ‘Marguerite’ cultivar strain of ‘Zairai Shiro.’ This primer ISSR-5 detected and identified certain specific band among the band pattern of ‘Southern Elegance White,’ this cultivar has cv. ‘Early White’ as the paternal parent. Similar polymorphic band pattern was confirmed by ISSR-PCR, in the breeding lines and the cultivars of the ‘Marguerite’ which have similarity in flower color, flowering type and share the hybrid parents. KEYWORDS: Argyranthemum, Glebionis carinata, Glebionis coronaria, ISSR, Marguerites, RAPD Argyranthemum is taxonomically placed in the Anthemideae, their commercial purposes. Asteraceae and grows sympatrically or allopatrically with Argyranthemum can be cross-hybridized easily with 24 species in Macaronesia in the Canary Islands of Spain Glebionis (Tsukamoto 1986; Ohtsuka and Inaba 2008). and in Madeira and the Salvage Islands of Portugal Up to the present, numerous hybrids among many (Bramwell and Bramwell 1974; Bremer and Humphries combinations of close relatives might have been occurred 1993). Argyranthemum is closely related to and easily artificially directly or indirectly to make cultivar makes hybrids with Glebionis (Trehane 1995), Ismelia complexity (Aoyama 2000) to perform multi-different and Heteranthemis (Bremer and Humphries 1993). A coloration and inflorescence heads. It was made question member of A. frutescens (L.) Schultz-Bip. has been well- that “What Marguerites are” according to his detection known as the common name of the ‘Marguerite’ group result by chromosome analysis of 38 cultivars and their with more than 100 cultivars imported to Japan from origin of so-called the commercialized A. frutescense that France since the Meiji Era (1868-1912) for the purpose of numerous closely related species and cultivars could be decolative ornamental plants in greenhouses and later cut- involved with hybridization and morphological types of flower products and potted flower products in the warmer cultivars from yellow to deep red-colored as well as white- sunny outdoor seashores step-by-step. Royal Horticultural colored ligulate as well as double flowers. Japanese Society of UK has listed more than 100 cultivars of cultivar. ‘Zairai Shiro’ showed the basic characteristics of ‘Marguerites’ (Cheek 1993). They have met so many white color-pigment and the ligulate flowers around the chances to face and hybridize with other genera of edge of the head inflorescence to perform the common chrysanthemums closely related to Argyranthemum character of the single flower head, but some species such during their long term cultivation mainly in Izu Peninsula, as A. maderense (D. Don) Humphries has originally natural Shizuoka Prefecture and Kagawa Prefecture, in Japan for yellow-colored flowers. Moreover, some ‘Margarite’ cultivars have been much more cross-hybridized to amplify *Based on the Master’s thesis of M. Morikawa submitted to Graduate diversifications by hybridization; for instance, ‘Margarite’ School of Agriculture, Tokyo University of Agriculture cv. ‘Izu Yellow’ might be a result of hybridization of 98 MORIKAWA ET AL. Glebionis coronaria to extract yellow color in the work of carinatum (Schousb.) Tzvelev were rather easily to 1970’s in the Izu area (Tsukamoto 1986). On the other hand, hybridize with Argyranthemum frutescence ‘Marguerites’ cv. ‘Margarites’ was cross-hybridized with G. carinata by (Zietkiewicz et al. 1994) since three species of Glebionis works of the Plant Cultivation and Breeding Section, Izu were closely related to the latter taxon. On the other hand, Agricultural Research Center, Shizuoka Prefectural 32 cultivars of ‘Marguerites’ were expected not only by Research Institute of Agriculture and Forestry, Izu breeders in France and other Europian nations but also Peninsula, Shizuoka Prefecture and made various hybrid many Japanese growers’ communities as well as staff cultivars such as ‘Canaria Queen,’ ‘Carnival Queen,’ members of the Plant Cultivation and Breeding Section, ‘Fu-Ren-Ka,’ ‘Garnet Queen,’ ‘Peach Queen,’ ‘Queen Izu Agricultural Research Center, Shizuoka Prefectural Mice,’ and so on as the member of ‘Margarite’ (Ohtsuka Recerch Institute of Agriculture and Forestry, Izu and Inaba 2008). Among them cv. ‘Fu-Ren-Ka’ was bred Agricultural Research Center, Izu Peninsula, Japan. and developed for some different purposes and different Additionally, 25 strains for new cultivars amplified were usefulness such as fragrance, decorative cut-flowers, studied here during the course of investigation by ISSR potmums (potted chrysanthemums), and so on and may (Inter Simple Sequence Repeat) and RAPD methods. bred to extract for many more different purposes. ISSR is a random primer and does not need the sequence Observing and concidering the appearances of such a big information for a primer design, and its operation is flower variant in the ‘Margarite’ could appear not only simple and easy like RAPD method and it is used for intraspecific hybridization within Argyranthemum phylogenetic analyses or identification of individual frutescence but also interspecific hybridization within the cultivars. It can be applied in many studies involving genus and some intergeneric hybridization. Thus, genetic identity, parentage, clone and strain identification, molecular analyses by RAPD by PCR and ISSR were and taxonomic studies of closely related species (Kondo applied to identify phylogenetic lineages (Williams et al. et al. 2003; Racharak and Eiadthong 2007). 1990). Comparing those amplified polymorphisms with DNA extractions. Total genomic DNA’s of the species RAPD can be used for identifying cultivars as well as of Argyranthemum and Glebionis, of their hybrids, 32 hybridities. The ISSR is amplified by using the sequence cultivars of Argyranthemum frutescence ‘Marguerites’ of an adjacent microsatellite region for a primer and 25 newly progressed strains for new cultivars studied (Zietkiewicz et al. 1994; Kondo et al. 2003; Tatarenko et were extracted from fresh leaves by using the CTAB al. 2013). Analyses of polymorphism with ISSR are method described by Doyle and Doyle (1987). Three partially inserted into the microsatellite region in the hundreds to 500 mg frozen leaf samples were homogenized genome. with liquid nitrogen and put in individual 2 ml tubes and Certain distinct characteristics of the flower heads of washed once with washing buffer [0.1 M Tris-HCl pH 8.0, the ‘Maguerites’ such as flower color in white, yellow, 2% 2-mercaptoethanol, 1% polyvinylpyrrolidone (PVP) pink and so on, single-ray flower-type, double-ray flower and 0.05 M ascorbic acid]. Two percent CTAB extraction type, anemone-type, inglewick tipped flower type, and buffer containing 2% 2-mercapthoethanol and 2% some other flower types and the fragrant-flower types polyvinylpyrrolidone were then added and incubated at should be originated not only from ‘Marguerites’ themselves 60°C for at least 30 min. After incubation, the samples but also from a lot of unrecorded interspecific cross- were centrifuged and the supernatant was collected. The hybridizations with many other species of Argyranthemum DNA was then isolated with 500µl 24:1 chloroform- by insect pollination, besides well-recorded man-made, isoamyl alcohol (CIA), precipitated with equal volume of hand pollinations. Thus, origin of numerous types of 2-propanol, and washed with 500 µl 24:1 chloroform- ‘Marguerites’ should be analyzed by molecular-based
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