胡杨杂种叶片组培技术研究

Study on Tissue Culture Technology of Hybrid Leaves of Populus euphratica

目的:优化胡杨杂种叶片组培技术进而为其遗传转化研究奠定基础。方法:以胡杨杂种叶片为外植体,以WPM为基本培养基,研究不同植物生长调节剂对叶片诱导愈伤组织、愈伤组织诱导不定芽、以及不定芽进一步诱导不定根的作用效果。结果:胡杨杂种叶片最佳脱菌方式:0.1%HgCl2脱菌5 min;叶片诱导愈伤组织的最适培养基为WPM+0.4 mg·L-16-BA+0.4 mg·L-1NAA,诱导率可达100%;愈伤组织诱导不定芽的最适培养基为WPM+0.5 mg·L-16-BA+0.2 mg·L-1NAA,诱导率可达100%;诱导不定根的最适培养基为1/2WPM+2.0 mg·L-1IBA+0.02 mg·L-1NAA+0.2 mg·L-1GA3,生根率可达100%。以上3种培养基均需添加30 g·L-1蔗糖,30 g·L-1琼脂,pH调至5.8。结论:本研究建立了以胡杨杂种叶片为外植体的组培技术,为后序进行胡杨杂种批量化生产及其遗传转化研究提供重要参考。

Objective:To optimize the tissue culture technique of hybrid leaves of Populus euphratica,and laid a foundation for the study of genetic transformation of P.euphratica.Method:The hybrid leaves of P.euphratica were considered as explants material,the effects of different plant growth regulators on callus induction,adventitious bud induction and adventitious root induction in the hybrid leaves of P.euphratica were studied base on the basic medium of WPM.Results:The optimum sterilization method for the hybrid leaves of P.euphratica was 0.1%HgCl2 for 5 minutes.The optimum medium for callus induction of the hybrid leaves of P.euphratica was WPM+0.4 mg·L-16-BA+0.4 mg·L-1NAA,and induction rate was 100%.For adventitious buds induction from the callus,the optimum medium was WPM+0.5 mg·L-16-BA+0.2 mg·L-1NAA,it exhibited 100%induction rate.The best suitable culture medium for adventitious roots induction was 1/2 WPM+2.0 mg·L-1IBA+0.02 mg·L-1NAA+0.2 mg·L-1GA3,rooting rate was up to 100%.The above three mediums should be supplemented with 30 g·L-1 sucrose,30 g·L-1 agarose and adjust pH to 5.8.Conclusion:In this study,the tissue culture technique with hybrid leaves of P.euphratica as explants was established,which provided an important reference for the study of mass production and genetic transformation of Populus euphratica hybrid.