NAA或潮霉素对小麦成熟胚愈伤组织分化的影响

Effects of NAA or Hygromycin on Differentiation of calli induced from mature embryo in Wheat

以5个栽培小麦品种诱导而来的成熟胚愈伤组织为外植体,研究了在分化培养基中加入NAA对小麦成熟胚愈伤组织分化的效果以及潮霉素浓度对不同阶段成熟胚愈伤组织分化的影响。研究结果表明,在对照的分化培养基中(含5mg/L激动素)加入0.1mg/L萘乙酸(NAA,1-naphthlcetic acid)形成的分化培养基,可显著提高4个供试小麦基因型的成熟胚愈伤组织的分化率,提高幅度达到12%～32%。不同的潮霉素浓度在"花培1号"愈伤组织的诱导阶段进行抗性筛选的结果表明,即使是在潮霉素浓度达到250mg/L时,仍有24%左右的愈伤组织存活率,这些存活的愈伤组织仍可能分化形成绿点甚至分化成苗,说明在"花培1号"愈伤组织的诱导阶段进行潮霉素抗性筛选是不适合的。5个不同小麦品种分化阶段进行潮霉素筛选具有明显的效果,但不同品种之间存在一定差异,其中:宁麦13的适宜潮霉素浓度为20mg/L,花培1号和扬麦158的适宜潮霉素浓度约为30mg/L,宁麦9和宁麦16的适宜潮霉素浓度在30～40mg/L左右。因此,我们认为,在分化阶段进行潮霉素抗性筛选是理想的筛选时期,在分化培养基中添加20～40mg/L浓度的潮霉素即可完全抑制小麦成熟胚愈伤组织的分化。本研究结果将有助于改进和完善普通小麦成熟胚遗传转化体系。

The effects on calli regeneration in regeneration medium with or without NAA and in different stage with different hygromycin concentration were studied by using the calli induced from mature embryo of five wheat cultivars as explants.The results showed the frequency of regeneration of calli induced from mature embryo in 4 wheat cultivars in the medium containing 5 mg/L KT＋0.1 mg/L NAA（KT,kinetin;NAA,1-naphthlcetic acid） was increased by 12%~32% more than those in the medium only containing 5 mg/L KT.There were 24% calli still survived when hygromycin concentration in the medium was up to 250 mg/L in callus induction stage in wheat of Huapei No.1.Some of the survived calli could also generate green spots or plantlets.This result suggested that it was not suitable for hygromycin resistance selection in callus induction stage of the wheat of Huapei No.1.However,the effect of hygromycin resistance selection in the regeneration stage of calli was obvious.Five cultivars had different resistance to hygromycin,the proper hygromycin concentration of selection was 20 mg/L for Ningmai13,30 mg/L for both of Huapei No.1 and Yangmai158,30~40 mg/L for both of Ningmai 9 and Ningmai16,respectively.The differentiation of calli induced from mature embryo was absolutely inhibited with 20~40 mg/L concentration of hygromycin,which suggested that the regeneration stage of callus was the best option for hygromycin resistance selection.The results of the study might be valuable and useful for improving the released cultivar which may be loss of some characters in the transgenic breeding programs of wheat.