赤霉素、光照及基因型对花生体细胞胚诱导和植株再生的影响

Effect of gibberellin,light and genotype on peanut somatic embryogenesis and plantlet regeneration

【目的】完善花生离体再生繁殖体系,为花生的遗传转化提供技术支持。【方法】以桂花系列花生品种为材料,以胚小叶为外植体,MS+10mg/L2,4-D为体细胞胚诱导基本培养基,MS+3mg/L6-BA+0.8mg/LNAA+(0～15mg/L)GA3为苗诱导基本培养基,研究光照、赤霉素和基因型对花生体细胞胚诱导和植株再生的影响。【结果】在光暗比14∶10条件下,花生体细胞胚诱导率明显高于全黑暗培养,体细胞胚诱导率增加7.5～37.5个百分点。在不同花生品种中,桂花26体细胞胚诱导率最高,达62.8%,桂花833最低,为21.7%。在成苗培养基中添加5～15mg/LGA3利于提高体细胞胚的成苗率。桂花26和桂花771在5mg/LGA3处理下成苗率最高,分别为42.8%和35.3%;桂花836、桂花1026、桂花833在15mg/LGA3作用下成苗率最高,分别为38.7%、33.3%、26.4%。所有品种与GA3组合中,以桂花26与5mg/LGA3组合成苗率最高,其次是桂花836与15mg/LGA3组合。【结论】适宜的光照条件有利于花生体细胞胚的发生;成苗培养基中添加GA3利于促进体细胞胚诱导成苗;桂花26和桂花836是体细胞胚诱导植株再生较理想的材料。

Objective Peanut in vitro regeneration system was optimized to provide technical support for its genetic transformation and mutagenesis. Method The effect of gibberellin, light and genotype on somatic embryogenesis and plantlet regeneration was studied using Guihua peanut varieties as materials, leaflets as explants, MS＋10 mg/L 2,4-D as induction medium, and MS＋3 mg/L 6-BA＋0.8 mg/L NAA＋（0-15 mg/L） GA 3 as germination medium. ResultThe frequency of embryo-genesis was significantly higher （light∶dark=14 h∶10 h） comparing with complete dark, and there was an increase range of percent point from 7.5 to 37.5 for different peanut varieties, respectively. The highest frequency of embryogenesis was 62.8% for Guihua 26, and the lowest was 21.7% for Guihua 833. The frequency of shoot development increased with a concentration range of GA 3 from 5 mg/L to 15 mg/L. The 5 mg/L of GA 3 was the best for Guihua 26 and Guihua 771 with a frequency of shoot development of 42.8% and 35.3% , respectively, and the 15 mg/L of GA 3 was the best for Guihua 836 and Guihua 1026 and Guihua 833. Their frequency of shoot development was 38.7%, 33.3% and 26.4%, respectively. Among all the combinations of variety and GA 3 concentration, the highest frequency of shoot development was Guihua 26 and 5 mg/L of GA 3 , and the next was Guihua 836 and 15 mg/L of GA 3 . Conclusion Appropriate light propels the development of peanut in vitro embryo. GA 3 can increase the frequency of shoot development from somatic embryos, and Guihua 26 and Guihua 836 are the best peanut varieties among the tested genotypes for somatic embryogenesis and plant regeneration.