Gürel Filiz, Karakaş Ozge, Albayrak Gülruh, Ari Sule
Department of Molecular Biology and Genetics , Istanbul University, Vezneciler, 34134 Istanbul Turkey.
Acta Biol Hung. 2009 Sep;60(3):309-19. doi: 10.1556/ABiol.60.2009.3.8.
In this study, induction of regenerable callus from mature embryos in eight Turkish barley varieties was analysed by using different plant growth regulators (PGRs). Varying concentrations (0.5-4 mg l -1 ) of 2,4-dichlorophenoxyacetic acid (2,4-D) and dicamba (3,6-dichloro-o-anisic acid) were tested for callus induction from mature embryos. Highest percent of callus induction was observed in Bornova 92 variety (98.3%) on MS medium supplemented with 4 mg l -1 dicamba. Calli were transferred to regeneration media with 0.5 mg l -1 dicamba, 0.5 mg l -1 zeatin riboside (ZR) and 2 mg l -1 thidiazuron (TDZ). Low concentrations of dicamba induced multiple shoots during callus regeneration. When the effect of precultivation with 2,4-D or dicamba on the shoot induction were evaluated, lower concentrations (< 4 mg l -1 ) of auxins have been found optimal. On the regeneration medium with 0.5 mg l -1 dicamba, shoots were able to elongate up to 20 cm and shoot numbers were between 1-23 per callus. The use of ZR led to formation of short shoot buds and somatic embryos in 2 weeks period. The effect of TDZ was different from other PGRs by inducing green solid sectors on calli surfaces (Total 51 sectors/20 callus/Akhisar variety). Five plantlets have been grown from these solid cell clumps and transferred to specific media for root formation. As a result, five varieties (Süleyman Bey, Bornova 92, Vamyk Hoca, Kaya and Akhisar) tested in our study showed the potential to produce regenerable callus by using low amounts of dicamba or TDZ. The optimization process starts from culturing embryos to plantlet formation took nearly 4 weeks.
在本研究中,通过使用不同的植物生长调节剂(PGR)分析了8个土耳其大麦品种成熟胚再生愈伤组织的诱导情况。测试了不同浓度(0.5 - 4 mg l⁻¹)的2,4 - 二氯苯氧乙酸(2,4 - D)和麦草畏(3,6 - 二氯邻茴香酸)对成熟胚愈伤组织诱导的影响。在添加4 mg l⁻¹麦草畏的MS培养基上,Bornova 92品种的愈伤组织诱导率最高(98.3%)。将愈伤组织转移到含有0.5 mg l⁻¹麦草畏、0.5 mg l⁻¹玉米素核苷(ZR)和2 mg l⁻¹噻苯隆(TDZ)的再生培养基上。低浓度的麦草畏在愈伤组织再生过程中诱导形成多个芽。当评估用2,4 - D或麦草畏预培养对芽诱导的影响时,发现较低浓度(< 4 mg l⁻¹)的生长素是最佳的。在含有0.5 mg l⁻¹麦草畏的再生培养基上,芽能够伸长至20 cm,每个愈伤组织的芽数在1 - 23个之间。使用ZR导致在2周内形成短芽和体细胞胚。TDZ的作用与其他PGR不同,它在愈伤组织表面诱导形成绿色固体扇形区(Akhisar品种,共51个扇形区/20个愈伤组织)。从这些固体细胞团中培育出了5株幼苗,并转移到特定培养基上进行生根。结果,在我们的研究中测试的5个品种(Süleyman Bey、Bornova 92、Vamyk Hoca、Kaya和Akhisar)显示出使用少量麦草畏或TDZ产生可再生愈伤组织的潜力。从培养胚到形成幼苗的优化过程耗时近4周。