Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana 141 004, India.
N Biotechnol. 2011 Jul;28(4):418-22. doi: 10.1016/j.nbt.2011.02.006. Epub 2011 Feb 23.
Tissue culture is one of the tools necessary for genetic engineering and many other breeding programs. Moreover, selection of high regenerating rice varieties is a pre-requisite for success in rice biotechnology. In this report we established a reproducible plant regeneration system through somatic embryogenesis. The explants used for regeneration were embryogenic calli derived from mature seeds cultured on callus induction media. For callus induction mature seeds were cultured on MS medium containing 30 g/l sucrose combined with 560 mg/l proline and 1.5-3.5 mg/l 2,4-D and 0.5-1.5 mg/l Kin. For plant regeneration, embryogenic calli were transferred to MS medium containing 30 g/l sucrose, supplemented with 1.0-3.0 mg/l BAP, 0.5-1.5 mg/l Kin and 0.5-1.5 mg/l NAA. The highest frequency of callus induction (44.4%) was observed on the MS medium supplemented with 2.5 mg/l 2,4-D, 0.5 mg/l Kin, 560 mg/l proline and 30 g/l sucrose. The highest frequency of shoot regeneration (42.5%) was observed on the MS medium supplemented with 2.0 mg/l BAP, 0.5 mg/l NAA and 0.5 mg/l Kin. The plantlets were hardened and transferred to soil in earthen pots. The developed method was highly reproducible. The in vitro developed plants showed normal growth and flowering under glasshouse conditions.
组织培养是遗传工程和许多其他育种计划所必需的工具之一。此外，选择高再生水稻品种是水稻生物技术成功的前提。在本报告中，我们通过体细胞胚胎发生建立了一个可重复的植物再生系统。用于再生的外植体是从成熟种子在愈伤组织诱导培养基上培养而来的胚性愈伤组织。用于愈伤组织诱导的成熟种子在含有 30 g/l 蔗糖的 MS 培养基上培养，同时添加 560 mg/l 脯氨酸和 1.5-3.5 mg/l 2,4-D 以及 0.5-1.5 mg/l Kin。对于植物再生，将胚性愈伤组织转移到含有 30 g/l 蔗糖的 MS 培养基上，补充 1.0-3.0 mg/l BAP、0.5-1.5 mg/l Kin 和 0.5-1.5 mg/l NAA。在补充有 2.5 mg/l 2,4-D、0.5 mg/l Kin、560 mg/l 脯氨酸和 30 g/l 蔗糖的 MS 培养基上观察到最高的愈伤组织诱导频率（44.4%）。在补充有 2.0 mg/l BAP、0.5 mg/l NAA 和 0.5 mg/l Kin 的 MS 培养基上观察到最高的芽再生频率（42.5%）。将幼苗硬化并转移到土陶盆中的土壤中。所开发的方法具有高度的可重复性。在温室条件下，体外发育的植物表现出正常的生长和开花。
