提高籼稻愈伤组织诱导率和增殖率的研究

Research of improvement of inducing and proliferation rate for rice(Oryza sativa L.subsp.indica)

以秋桂矮11、华籼占、万籼占45等3个籼稻品种的种子为外植体,消毒后接种在不同培养基上,(25±1)℃下暗培养20～30天后比较不同品种愈伤组织的诱导率,再将愈伤组织继代培养20天后调查其增殖率.结果表明,在NMB培养基中添加2.5～3.0 mg/L 2,4-D有利于提高籼稻愈伤组织诱导率,添加脯氨酸、水解酪蛋白等营养物有利于提高万籼占45的愈伤诱导率,但对提高秋桂矮11、华籼占的愈伤诱导率影响不大;在NPCS培养基(NMB+0.5g/L 脯氨酸+0.3 g/L水解酪蛋白+3～5 g/L 山梨醇)继代培养1～2次,以30 g/L麦芽糖为碳源、2,4-D浓度为2.5 mg/L时,籼稻愈伤组织活性明显提高,其胚性愈伤组织的增殖率最大.

The seeds of three indica rice （Oryza sativa L. subsp. indica） Qiuguiail 11 ,Huaxianzban and Wanxianzban 45 were collected. Those were carried out on different basal media after sterilization treatment, several media with different combination of growth regulators were tried. After 20-30 days of incubation under dark at 25 ± 1℃, calli initiated from Scutellum were subcultured on fresh callus induction media. After 20 days, Systemic researches bad been carried for callus inducing ratio and the maximum proliferation efficiency of emhryogenic callus. High frequency induction from mature seeds derived embryogenic callus was initiated on NMB combination medium supplement with 2.5 - 3.0 mg/L 2,4 - D, for nutrient substance such as proline,casein acid hydrolysate and D - sorbitol, it is beneficial only for Wanxianzban45. Those calli were sub cultured in multiplied media, in NPCS medium （basal medium NMB＋0.5 g/L proline ＋ 0.3 g/L CH＋ 3-5 g/LD-sorhitol） with 30 g/ L maltose and 2.5 mg/L 2,4 - D, the multiplied vigorously callus and the maximum proliferation of embryogenic callus were obtained.