Verma Dipti, Joshi Rohit, Shukla Alok, Kumar Pramod
Department of Plant Physiology, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar 263 145, India.
Indian J Exp Biol. 2011 Dec;49(12):958-63.
Development of highly efficient and reproducible plant regeneration system has tremendous potential to provide improved technology to assist in genetic transformation of indica rice cultivars for their further exploitation in selection. For the development of a highly reproducible regeneration system through somatic embryogenesis, mature embryos of highly popular rice cultivars i.e., Govind (for rainfed areas), Pusa Basmati-1 (aromatic basmati) and Jaya (for irrigated areas) were used. Optimum callus formation (%) to MS medium supplemented with 2, 4-D was obtained at 12.0 microM in Govind, 14.0 microM in Jaya and 15.0 microM in Pusa Basmati-1. All the cultivars showed good proliferation on MS medium without hormone. In Govind, highest embryogenic response was observed in MS medium supplemented with 2, 4-D (0.4 microM) + kinetin (0.4 microM), while in Pusa Basmati-1 with 2, 4-D (0.4 microM) + kinetin (2.0 microM) and in Jaya on hormone-free MS medium. Excellent embryo regeneration in Govind was observed on MS medium supplemented with low concentrations (1.1 microM) of BAP or hormone-free MS medium, while in Pusa Basmati-1 and Jaya embryogenesis was observed on MS medium supplemented with higher concentration of BAP (2.2 microM). Similarly, maximum plantlets with proliferated roots were observed in Govind on hormone-free MS medium, while in Pusa Basmati-1 and Jaya on MS medium supplemented with high concentration of NAA (4.0 microM). Developed plantlets were further successfully acclimatized and grown under pot culture up to maturity. Further the yield potential of in vitro developed plants was accessed at par to the direct seeded one under pot culture. Present, protocol standardizes somatic embryogenesis and efficient regeneration of agronomically important, high yielding and diverse indica rice cultivars which can be utilized as an efficient tool for molecular studies and genetic transformation in future.
高效且可重复的植物再生系统的开发具有巨大潜力，可为籼稻品种的遗传转化提供改进技术，以便在选育中进一步加以利用。为了通过体细胞胚胎发生开发高度可重复的再生系统，使用了广受欢迎的水稻品种的成熟胚，即Govind（用于雨养地区）、Pusa Basmati - 1（香米巴斯马蒂）和Jaya（用于灌溉地区）。在添加2,4 - D的MS培养基上，Govind在12.0微摩尔浓度下获得最佳愈伤组织形成率（%），Jaya在14.0微摩尔浓度下，Pusa Basmati - 1在15.0微摩尔浓度下。所有品种在无激素的MS培养基上均表现出良好的增殖。在Govind中，在添加2,4 - D（0.4微摩尔）+激动素（0.4微摩尔）的MS培养基中观察到最高的胚性反应，而在Pusa Basmati - 1中为2,4 - D（0.4微摩尔）+激动素（2.0微摩尔），Jaya则在无激素的MS培养基上。在添加低浓度（1.1微摩尔）BAP的MS培养基或无激素的MS培养基上观察到Govind有优异的胚再生现象，而在Pusa Basmati - 1和Jaya中，在添加较高浓度BAP（2.2微摩尔）的MS培养基上观察到体细胞胚胎发生。同样，在无激素的MS培养基上Govind观察到最多的生根增殖小植株，而在Pusa Basmati - 1和Jaya中，在添加高浓度NAA（4.0微摩尔）的MS培养基上观察到最多。发育的小植株进一步成功驯化，并在盆栽条件下生长至成熟。此外，在盆栽条件下，对离体培养发育的植株的产量潜力与直播植株进行了同等评估。目前，该方案规范了体细胞胚胎发生以及农艺性状重要、高产且多样的籼稻品种的高效再生，未来可将其用作分子研究和遗传转化的有效工具。
