Mariashibu Thankaraj Salammal, Subramanyam Kondeti, Arun Muthukrishnan, Theboral Jeevaraj, Rajesh Manoharan, Rengan Sampath Kasthuri, Chakravarthy Rajan, Manickavasagam Markandan, Ganapathi Andy
Department of Biotechnology and Genetic Engineering, School of Biotechnology, Bharathidasan University, Tiruchirappalli 620 024, India.
Indian J Exp Biol. 2013 Oct;51(10):849-59.
Majority of the Indian soybean cultivars are recalcitrant to tissue culture regeneration. The present communication reports the development of somatic embryogenesis in a liquid culture medium from immature cotyledons of G. max. Following induction with 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthalene acetic acid (NAA), the number of somatic embryos and percentage of explants that responded were higher with 45.24 microM 2,4-D. The proliferation of somatic embryos for three successive cycles was achieved in 22.62 microM 2,4-D. Histodifferentiation of somatic embryos under NAA (10.74 microM) indicated that better embryo development and maturation was achieved without any growth regulator. The amino acids such as L-glutamine favoured the somatic embryo induction and histodifferentiation at 20 and 30 mM respectively, where as L-asparagine at 10 mM concentration enhanced the somatic embryo proliferation. In addition, somatic embryos that were desiccated (air-drying method) for 5 days showed better germination (40.88%). The Indian soybean cultivars also showed strict genotypic influence and cv. Pusa 16 was emerged as a best responding cultivar for somatic embryo induction with 74.42% of response.
大多数印度大豆品种对组织培养再生具有顽拗性。本通讯报道了从大豆未成熟子叶在液体培养基中体细胞胚胎发生的进展。在用2,4-二氯苯氧乙酸(2,4-D)或萘乙酸(NAA)诱导后,45.24微摩尔/升的2,4-D处理下体细胞胚的数量和外植体的响应率更高。在22.62微摩尔/升的2,4-D中实现了体细胞胚连续三个周期的增殖。在NAA(10.74微摩尔/升)下体细胞胚的组织分化表明,在没有任何生长调节剂的情况下能实现更好的胚发育和成熟。氨基酸如L-谷氨酰胺分别在20和30毫摩尔时有利于体细胞胚诱导和组织分化,而10毫摩尔浓度的L-天冬酰胺促进体细胞胚增殖。此外,经5天干燥处理(空气干燥法)的体细胞胚表现出更好的发芽率(40.88%)。印度大豆品种也表现出严格的基因型影响,品种Pusa 16成为体细胞胚诱导响应最佳的品种,响应率为74.42%。