توليد جنين هاي هاپلوئيد از طريق كشت ميكروسپور درگندم ( TRITICUM AESTIVUM L )

The Production of Haploid Embryos via Microspore Culture of Wheat (Triticum aestlvum L.)

In this study, eight commerical Iranian wheat cultivars were tested for their microsporc culture response. Spikes aseptically pretreated in strileinducer formulation [O.lg L-1 of2-hydroxynicotinic acid, 10-6 mol L -1 2,4-0 and 10-6 mol L41 BAP) and placed in incubator at 33"Cfor II reguired period of time, ranging between about 48 and72 h. Microspores were isolated through blending of spikes in rnicrospore is elation soiuting (OJ mol L"\ mannitol). Isolated microspores were cocultured with immature ovaries in liquid NPB99 medium at a density of approximately 4xl03 rnicrospores mL·1 and incubated in the dark and at 27°C. After 3-4 weeks, response of wheat cultivars for embryogenesis WII,s evahraed, Cytological investigations using inverted microscope showed thatmulticellular structures, were formed inapproximately one week after culture. During the second week, themicrospore wall ruptured and immature embryoids emerged which grew into mature embryos within about 10 to 14days. Analysis of variance indicated significant differences among cultivars for embryogenesis, thatdemonstrates the genetic dependence of thistrait.Cuitivars of Falat and MoghanI with means of 1427±81.58 and 1108±54.55 embryos perspike respectively, were identified as the mostsupperior cultivares inmicrcspore culture.