相比粳稻而言,籼稻愈伤组织诱导一直没有一种通用的诱导培养基。为了找到适用于籼稻愈伤诱导的通用培养基,以明恢63、IR64、RD23三个籼稻品种的成熟胚为材料,在固定成分(300 mg/L水解酪氨酸蛋白+500 mg/L脯氨酸+250 mg/L谷氨酰胺+30 g/L蔗糖+8 g/L琼脂)和相同的培养条件下,以基本培养基(MB、NB、N6和CC 4种基本培养基)、2,4-D(2.0,3.0,4.0、5.0 mg/L 4种浓度)、6-BA(0,0.2,0.4,0.5 mg/L 4种浓度)为影响因素,利用正交实验设计出16种培养基组合,通过评价愈伤质量和愈伤诱导率来筛选最适合的籼稻愈伤组织诱导培养基。结果表明,2,4-D能提高愈伤诱导率,不用6-BA时愈伤质量最好,基本培养基MB的愈伤诱导效果最好。筛选出的籼稻愈伤诱导通用培养基是MB+2,4-D 5.0 mg/L,在28℃黑暗条件下培养20 d。用筛选出来的通用培养基对不同来源的籼稻品种进行愈伤组织诱导,结果表明该通用培养基对多数籼稻品种具有稳定的愈伤诱导效果。
In order to find a general inducing medium for cultivars of Indica rice,an orthogonal text containing three factor,which were the basic medium(NB,N6,CC and MB),2,4-D(2.0,3.0,4.0 and 5.0 mg/L)and 6-BA(0,0.2,0.4 and 0.5 mg/L) was designed.There were 16 medium combinations based on same nutrient components(300 mg/L casein hydrolysate + 500 mg/L proline + 250 mg/L glutamine + 30g /L sucrose + 8g/L agar) with three cultivars of Indica rice,which were Minghui63,IR64,and RD23,were employed for inoculation on the 16 medium combinations and culture in the same conditions.The callus induction rate and quality of each combination were evaluated.The results indicated that the 2,4-D significantly increased callus inducing rate,the 6-BA significantly decreased the quality of callus and the MB medium was the most suitable basic medium for callus inducing.The general medium for callus inducing for cultivars of Indica rice was the combination including MB medium,5.0 mg/L 2,4-D.The culture condition for induction was in the dark for 20 days with 28 ℃.The confirmed experiment showed that the medium was suitable for most cultivars of Indica rice.