Narcissus pseudonarcissus, a flowering bulbous plant of the Amaryllidaceae family, is a typical Mediterranean genus of geophyte, with unique flower shape (with cup-shaped or crown-like corona) and outstanding flower color (Fernandes, 1968). In recent years, growing attention has been focused on N. pseudonarcissus, owing mainly to its horticultural success and popularity and the abundant alkaloids in bulbs with important biological activities (Bastida et al., 2006).
There are thousands of cultivars of Narcissus registered during the past century. Among them, many are natural hybrids or natural interspecific hybrids. As the members of the genus are so popular in cultivation and have been hybridized over such a long period, a complex range of hybrid forms occur with unknown state of the parents’ genetic characteristics. It is now difficult or impossible to elucidate the exact parents for most existing hybrids (Fernández, 1984; Marques et al., 2010; Ribeiro et al., 2007). Therefore, artificial hybridization with distinct breeding objective on N. pseudonarcissus is still relatively backward development for the following reasons:
1) The chromosome number is usually 2n = 14 or 28 (Philp, 1934) for the species N. pseudonarcissus, but there is considerable variation throughout the genus (Darlington and Janaki, 1945), and aneuploidy and polyploidy are quite common. In addition to some diploids, most types face the certain problem of fertility decrease in various degrees or even sterility (Brandham, 1992). 2) N. pseudonarcissus, as a perennial bulbous geophyte, mainly reproduces by developing vegetative daughter bulblets. The seedling plant is usually very small during the first 4 years and only has vegetative growth and first flowers in the fifth or sixth season of growth if survived from insect visitation every year (Caldwell and Wallace, 1955). This makes breeding quite inefficient, expensive, and time-consuming. 3) Breeding is also being impeded because of the asynchronous nature of flowering in N. pseudonarcissus. To conduct successful crosses, it is necessary to select genotypes on their individual values and to plant them in a way that will enable flowering at almost the same time. 4) The cultivars chosen as parents are always not homozygous in most genetic loci. Genetic segregation will be comprehensive in the unbeknown genetic background and molecular breeding still has a far way to go. Moreover, low fertility and disease also affect the obtaining of artificial hybrids. All those mentioned previously hindered the efficiency of crosses to obtain new N. pseudonarcissus cultivars with desirable traits. Although hybridization, polyploidization, and adaptive radiation occur within the genus along with evolutionary processes, there were no artificial genetic populations with clear genetic background, and no F1 hybrids generation has been reported as far as we know.
Over the past decades, molecular markers, especially RAPD markers, have been extensively used with considerable success in several horticultural plants, including lemon (Deng et al., 1995), fig (Galderisi et al., 1999), strawberry (Hokanson et al., 2000), pecan (Conner and Wood, 2000), and apricot (Badenes et al., 2000) for germplasm characterization, population structure analysis, and assessment of genetic diversity. Despite the economic and ecological importance of N. pseudonarcissus, little genomic information is available for this genus, and molecular markers have only recently been used in genetic studies. In addition, both basic genetic researches related to population studies and prebreeding programs of daffodil remain scarce for most Narcissus species. RAPD was used to analyze the genetic structure of populations of N. pseudonarcissus L. at the regional level (Guy et al., 2010) and the genetic variations of Narcissus tazetta var. chinensis (Chen et al., 2002). There were no reports of molecular markers used in some specific genetic populations in Narcissus.
Narcissus pseudonarcissus cultivar Slim Whitman was bred by the Netherlands breeder D.P. de Graaf and first flowered at 1978. ‘Pinza’ was registered in 1962, and it was popular and won a lot of awards since then (http://daffseek.org/ and http://apps.rhs.org.uk). They are famous and widely cultivated varieties all the time for their attractive flowers: ‘Slim Whitman’ with white perianth and light yellow corona, ‘Pinza’ with yellow perianth and yellow-orange corona. They are both large-cupped daffodils and possess the same basic chromosome numbers (2n = 28) (Brandham, 1992). Most importantly, they are fertile, which means they can be used as parents in cross breeding. In this study, we did reciprocal crosses between ‘Slim Whitman’ and ‘Pinza’, aiming to select individuals with novel flower color or flower shape in the F1 generation. The morphological and molecular differences of the reciprocal cross hybrids were further analyzed to elucidate the inheritance patterns on different traits and to facilitate the breeding development of new market-oriented daffodil flowers.
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