Biotech Studies 2016, Vol 25, Num, 1     (Pages: 237-242)

Obtaining the Pure Line in F2 Generation Wheat Using Anther Culture Technique

Özcan YORGANCILAR 1 ,Aysel YORGANCILAR 1 ,Serhat DİKMEN 2 ,Serdar DİKMEN 2 ,Merve ÇARIKÇI 1 ,Fikret EVCEN 2 ,Fahriye VAN 2 ,Pervin UZUN 1 ,Aysen YUMURTACI 3 ,İmren KUTLU 4 ,Zeynep SİREL 1

1 Geçit Kuşağı Tarımsal Araştırma Enstitüsü Enstitüsü Müdürlüğü, Eskişehir
2 Dikmen Tarım Ürünleri Limited Şirketi Söğüt, Bilecik
3 Marmara Üniversitesi Biyoloji Bölümü, İstanbul
4 Osmangazi Üniversitesi Ziraat Fakültesi, Eskişehir
DOI : 10.21566/tarbitderg.280498 - This study was conducted at theTransitional Zone Agricultural Research Institute in the years 2013- 2014.The purpose of the study was providing the combination conventional breeding in tegration with anther culture techniques, so obtaining more than %100 pureline in a short time according to conventional breeding methods, saving time andreducing cost for development of new varieties. In this research, 61 genotypes of F2 hybrid bread wheat were used. F2 hybrid seeds were sown in greenhouse in August 2014, after vernalization requirement was satisfied. Grain ears were taken at October, 2014. Ears which were taken at early-midsingle-coreperiod, were wrapped in plastic bags at 4°C and kept during the 12-15 days.After ears were made sterile, anther cultivation was made 50-100 units per each petri dish, in the MN6 nutrient media.Petri dishes which were containing the anthers were left in the incubator at dark conditionsand 28°C. Callus which were formed after 28 days in the incubator, was transferred directly into a sterile cabine and inserted in 190 II solid regeneration medium. The petri dishes which callus`s were transfered into, were kept at 25°C for 16 hours light (50 μmol s-1 m-2) and 8 h dark conditions at the climate chamber until they regenerated. Growing plants In the semedia were transferred to the190 II medium containing stem test tube when they were done 1-1.5 cm. As a result, 129.744 anther was planted from F2 hybrids, 14,836 callus were formed and 1104 plants were obtained. At the continuation of work, the obtained plants began to be transferred into the cold climate chamber at 4°C for the vernalisation. Vernal plants were transferred into pots at 15°C for 16 h light / 8 hours of darkness for waiting two weeks in the climate cabinet. Ploidy level of plants were determinated and the spontaneous diploid ones will be transferred to the greenhouse and climate cabinets. Colchicine will be applied to the haploid plants for chromosome doubling. Then, the obtained seeds from the pure lines will be used for the seed multiplication. Keywords : Anther, double haploid, wheat