Echinacea Purpurea) for Genetic Enhancement of Cultivated Sunflower (Helianthus Annuus L.
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Turkish Journal of Agriculture and Forestry Turk J Agric For (2014) 38: 15-22 http://journals.tubitak.gov.tr/agriculture/ © TÜBİTAK Research Article doi:10.3906/tar-1210-91 Utilization of related wild species (Echinacea purpurea) for genetic enhancement of cultivated sunflower (Helianthus annuus L.) Roumiana VASSILEVSKA-IVANOVA, Boris KRAPTCHEV*, Ira STANCHEVA, Maria GENEVA, Ivan ILIEV, Georgi GEORGIEV Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Sofia, Bulgaria Received: 02.11.2012 Accepted: 13.05.2013 Published Online: 13.12.2013 Printed: 20.01.2014 Abstract: Helianthus annuus is one of the most important oil species, with a comparatively narrow genetic base. The development of new sunflower cultivars is the focus of many research and breeding programs. Intergeneric hybridization betweenH. annuus and Echinacea purpurea, a valuable medicinal plant, has not yet been utilized in cultivar development. This paper describes 2 advanced hybrid lines produced from an intergeneric cross between the cultivated sunflower inbred line 6650 and an accession of E. purpurea. The hybrid plants have successfully been grown in the field. The hybrids, showing expression of traits from both parental species, were intermediate between both parents for plant height, leaf shape, leaf color, floral characteristics, tocopherol content, and fatty acid profile. It could be postulated that the hybridization Helianthus × Echinacea offers opportunities for combining desirable traits that increase the variability of cultivated sunflower. Key words: Antioxidant content, cultivated sunflower, Echinacea purpurea, fatty acid, Helianthus annuus, wide hybridization 1. Introduction is evidence that intentional crosses between species in Wide hybridization, including both interspecific and different genera have the potential for crop improvement intergeneric hybridization, is one of the most important and to increase the genetic variability of crop species, strategies for creating variations in plant species since it including for H. annuus in particular. It is plausible that has the potential to combine useful traits, i.e. favorable gene transfer between members of different genera may morphology, disease resistance, and some environmental complement the more classical hybrids that provide gene tolerances of both parents that could not be achieved by flow across interspecific crosses. Additionally, they have crossing within a single species. Sunflower (Helianthus proved to be very useful for elucidating the evolutionary annuus L.) cultivars lack acceptable levels of genetic relationships between H. annuus and related wild species variation since hybrid breeding utilizes a comparatively of Asteraceae. narrow genetic base. Wild Helianthus species are a potential The present work is a part of a sunflower research source of genes for resistance or tolerance to insects, program with the objective of producing and evaluating disease, and pests; for early maturation and resistance in new interspecific or intergeneric hybrids that provide unfavorable environmental factors (soil salinity, acidity, novel combination of traits useful for plant breeding. and drought); and as sources of cytoplasmic male sterility In particular, the program combines interest from both and fertility restoration (Faure et al. 2002; Seiler and Gulya plant breeding and academic research in the use of wide 2004). Successful interspecific transfer of traits from wild hybridization for transferring desirable traits from wild species to cultivated sunflower was a reason for attempts relatives to cultivated sunflower, developing germplasm of wider crosses, including those between members of pools having wild Helianthus genes in a domestic related genera. background, and for characterizing phylogenetic affinity. Advances in intergeneric crosses in tomato (Rick et al. Hybrids between cultivated H. annuus and species of 1986), species of wild grasses from the genera Aegilops, different genera of the family Asteraceae such as Tithonia Agropyron, and Secale (Fedak 1984); wheat; rapeseed rotundifolia and Verbesina encelioides, identified as having (Brassica napus); and tobacco (Nicotiana tabacum) have intermediate characteristics, have been reported in our been well documented (Goodman et al. 1987). There previous studies (Christov and Vassilevska-Ivanova 1999; * Correspondence: [email protected] 15 VASSILEVSKA-IVANOVA et al. / Turk J Agric For Vassilevska-Ivanova et al. 1999; Vassilevska-Ivanova and protected from foreign pollen by bagging. First-generation Tcekova 2005; Vassilevska-Ivanova 2005; Vassilevska- hybrid plants were verified using morphological and Ivanova et al., 2013). cytological methods, and F1 hybrids were back-crossed Herewith we present data on the wide hybridization to common sunflower to obtain BC1 and BC2. Some BC2 between cultivated sunflower and Echinacea purpurea (L.) progeny possessed a new phenotype (light green dwarfed Moench, one of the coneflowers of the genus Echinacea. plants with specifically serrated leaves) that was not Coneflowers are a group of native American wildflowers observed in the parents. These traits were selected for and from the daisy family (Asteraceae) characterized by spiny fixed after 2BC to produce 2 introgression lines (branched flowering heads with an elevated receptacle that forms the and unbranched). Seeds from advanced plant generations “cone”. We chose Echinacea purpurea for several reasons: were produced after self-pollination under a bag. The 1) it has a high medicinal value worldwide, resulting from plant growth conditions that were employed have already the combined effects of several phytochemicals; 2) as a been described (Vassilevska-Ivanova and Naidenova model of wide hybridization between 2 distantly related 2005). Both lines represented here are F8 progeny of selfed genera of Asteraceae, its successful cross with sunflower plants; they were raised in the experimental field of the would be of substantial benefit for plant science and plant Institute of Plant Physiology and Genetics, Sofia, Bulgaria breeding; 3) the intriguing aspect of wide hybridization as (42°50′N, 23°0′E, altitude of 595 m) during 2010–2012. a powerful tool for the development of new useful genetic Seeds were sown in 5-m-long rows spaced 0.70 m apart resources and crop improvement; and 4) modern breeding with 3 replications. The sowings were performed in mid- has renewed the interest for wide hybridization-derived April. Conventional management practices were used. materials with molecular assisted selection to release Morphological traits were recorded for 30 random plants adaptive and high-yielding cultivars. from both lines and their parents. The methods used in There are 9 species of Echinacea, all of which are this investigation for hybridizing plants, fertility tests, perennials. Being largely used in traditional medicine, and morphological comparisons are the same as those E. pallida, E. purpurea, and E. angustifolia have been described in previous reports (Vassilevska-Ivanova and the most investigated of these species. E. purpurea, or Tcekova 2005). “purple” coneflower, is a clump-forming herbaceous plant 2.2. Phenotype observations with chromosome number 2n = 22. The daisy-like flower Thirty randomly selected plants from each line and both heads are very attractive with rose-purple rays and large, parents were used for recording days to flower (50%), self- cone-shaped purple-brown centers (American Botanical compatibility (%), head diameter (cm), branched or not Council, 1994; Greenfield and Davies, 2004). branched, stainability of pollen using the acetocarmine This paper reports on 2 intergeneric Helianthus × test, 1000-seed weight (g), absolute dry matter (%), and Echinacea hybrid lines produced by conventional crosses. seed protein content (N × 6.25) calculated by the standard Their distinguished phenotype and agronomic traits as method. Morphological floral characteristics included well as antioxidant potential and fatty acid composition number of ray flowers, length of the corolla of ray flowers indicated that these lines have the potential to provide (cm), and width of the corolla of ray flowers (cm). All insight into the processes and consequences attendant floral characteristics were measured at the end of anthesis. to wide hybridization. Strategies for utilizing members 2.3. Antioxidants analyses of different genera in conventional sunflower breeding The total antioxidant capacity (free radical scavenging programs for crop improvement are discussed. activity) as well as the presence of ascorbate and tripeptide thiol, glutathione (α-glutamyl cysteinylglycine), 2. Materials and methods tocopherols, phenols, and flavonoids was determined in 2.1. Plant material leaves from 10 plants (bulked probe) of the hybrid lines Both intergeneric lines originated from pollination and parents. All methods used were previously described of cytoplasmic male sterile line L 6650 of cultivated (Stancheva et al. 2011). The ascorbate, glutathione, and sunflower Helianthus annuus (female), released by the tocopherol content were assayed spectrophotometrically Dobrudja Agricultural Institute, Bulgaria, with bulked using a slightly modified method based on absorption of pollen from wild perennial Echinacea purpurea (male). a phosphomolybdenum complex proposed by Prieto et al. The population of E. purpurea used in this study was (1999). The total antioxidant capacity was measured by recovered from the genetic collection of the Institute bleaching