Garden stock (Matthiola incana R. Br.) is a commercially important horticultural crop having different types of flowers varied in color and flowered phenotype, belonging to the family Brassicaceae (Tatsuzawa et al. 2012).
Due to high ornamental value, many studies associated in culture and breeding of the garden stock have focused on improvement of morphological and physiological traits such as flower color, double flower phenotype, inflorescence length, and fertility (Sima 2019a, b; Daozong et al. 2018).
There are single and double flowered M. incana cultivars, and latter is more widely used for floriculture because it has more beautiful flowers. Seed production is only possible in single flowered individuals. It is impossible in double flowered plants due to degenerative pistils and stamens.
Therefore many researchers and breeders have aimed to produce of more double flowered plants than single flowered ones in M. incana and to develop markers to select them.
To date, several morphological traits such as cotyledon shape, serrate leaf, seed color, and leaf color have been used in early selection of double flowered stock individuals, because these traits were linked to double flowers (Saunders 1911; 1915; 1921; 1928; Ecker et al. 1993).
Takashi and Kanae (2018) identified the gene related to double flowers and developed a molecular marker to select double flowered plants efficiently, based on the study of double flower-linked molecular markers in Japanese gentian (Tasaki et al. 2017).
In general, the double flower trait is inherited by a single locus, s and it is linked to leaf color. In some stock cultivars, leaf color is a simple morphological marker, that is, green color indicates single flowered individual while pale green color represents double flowered individual (Ecker et al. 1993).
And also the fertility of plant is an important trait in cultivation and breeding of stock as well as other plants. Consequently, it is necessary to make a good stock cultivar with high fertility, different colors, and being capable of early discrimination of double flowered phenotype, in culture and production for commercial use.
Random amplified polymorphic DNA technique has been widely applied to evaluate genetic diversity on intraspecific level such as Ocimum (Lamiaceae) (Tanmay et al. 2016) and Swertia (Gentianaceae) (Prabhjot et al. 2019) and interspecific level including Citrullus colocynthis (Kumar et al. 2017), Lactuca sativa L. (Shubhangi et al. 2018), Nilgirianthus ciliates, Artemisia herba-alba (Khaled et al. 2019).
Moreover, RAPD method has also been used to assess genetic diversity on the level of variety or cultivar in following plants, for example, common beans (Phaseolus vulgaris L.) (Maciel et al. 2001), ginger (Siddharth et al. 2007), apricot (Prunus armeniaca L.) (Chroboková et al. 2011), cowpea (Masvodza et al. 2014), Prosopis cineraria (L.) Druce (Palaiyur et al. 2016), flax (Linumu sitatissimum L.) (Rozhmina et al. 2016), and Penthorum chinense Pursh (Zhiqiang et al. 2017).
And also ISSR method has been widely used in genetic diversity analysis because of its high reproducibility than RAPD method (Domenyuk et al. 2002; Galvan et al. 2003; Dogan et al. 2007).
However, in stock (M. incana), there are few reports on the study using molecular markers (Amaal 2009; Bekir et al. 2016; Takashi and Kanae 2018).
The present work aimed to: (1) make superior cultivars with different colors, high fertility, being capable of early selecting only double flowered seedlings by leaf color and examine morphological characteristics and (2) select RAPD and ISSR primers for cultivar certification and identification to culture and maintain good commercial stock cultivars.
The white cultivar is highly fertile, but it is not possible to distinguish between single flowered individuals and double flowered ones. Otherwise, in case of the pink cultivar, although it shows very low fertility, it is possible to select double flowered seedlings according to leaf color; single flower phenotype with green color and double flower phenotype with pale green color.
Here we evaluated genetic diversity among new double flowered stock cultivars obtained in cross “white” cultivar × “pink” cultivar by using morphological traits including leaf color, leaf shape, flower color and double flower phenotype, and RAPD and ISSR markers.
In this study we made some good stock cultivars with different colors including pink, pale pink and white (high fertile and capable of distinguishing between single and double flowered seedlings) through outcrossing between white cultivar (high fertile but impossible to select double flower phenotype) and pink cultivar (vis. versa). In newly obtained stock cultivars, it is possible to distinguish between single and double flower seedlings by leaf color, having about 70 ~ 75% of fertility, thereby they are efficient good cultivars than both parent cultivars in seed production and culture.
Additionally, our cultivars were classified into two main groups, including the pink and the white cultivars respectively, the genetic distances among them varied from 0.03 to 0.24, with similarity to the previous research results in apricot and cowpea (Chroboková et al. 2011; Masvodza et al. 2014).
In the future, by using our selected RAPD and ISSR primers, the present method might be contributed to establish the cultivar certification, identification, and conservation system in culture and management of double flowered stock cultivars with high fertility and different colors.