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An efficient in vitro regeneration protocol for faba bean (Vicia faba L.)

机译:一种高效的蚕豆体外再生方案(Vicia faba L.)

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A reliable regeneration system for faba bean (Vicia fabaL.) has been difficult to establish, delaying its genetic improvement. In the present work a rapid, reproducible and efficient regeneration method was developed for faba bean using single cotyledon explants with half embryonal axis. MS medium supplemented with 6 µM TDZ (thidiazuron), 10 µM 2-iP and 4 µM kinetin induced 30 to 50 adventitious buds /shoots after two weeks of culture, which were elongated on MS medium supplemented with 6 µM 2-iP and 2 µM kinetin. With healthy and strong roots established, shoots were transferred to MS medium supplemented with 5 µM IBA within 10 to 14 days. Shoots of 5 cm long were most suitable for rooting. Potting-mixture with good aeration and less capacity to retain water was most suitable for successful establishment of faba bean plantlets. Garden soil mixed with sand (gravel) and bio-manure (1:1:1) was most suitable for transplantation. TDZ promoted adventitious bud formation while 5 µM IBA was most suitable for rooting, higher concentrations were toxic to plantlets. Aeration of the potting mixture was important for rapid micropropagation and successful establishment. The efficient regeneration protocol reported here allows for successful micropropagation of faba bean, which is essential for future genetic improvement of plants via transformation protocols.
机译:难以建立可靠的蚕豆再生系统(Vicia fabaL。),延缓了其遗传改良。在目前的工作中,使用半胚轴的单个子叶外植体开发了一种快速,可重现和高效的蚕豆再生方法。培养两周后,添加了6 µM TDZ(噻唑隆),10 µM 2-iP和4 µM激动素的MS培养基诱导了30至50个不定芽/芽,并在添加6 µM TD的MS培养基上延长M 2-iP和2μM激动素。建立健康而牢固的根系后,将芽在10到14天内转移到补充了5 µM IBA的MS培养基中。 5厘米长的芽最适合生根。具有良好通气性和较低保水能力的灌装混合物最适合成功建立蚕豆苗。混合有沙子(砾石)和生物肥料(1:1:1)的花园土壤最适合移植。 TDZ促进不定芽的形成,而5 µM IBA最适合生根,更高的浓度对幼苗有毒。灌注混合物的充气对于快速微繁殖和成功建立非常重要。此处报道的高效再生方案可以使蚕豆成功进行微繁殖,这对于通过转化方案对植物进行未来的遗传改良至关重要。

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