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目的蛇足石杉Huperzia serrata自然繁殖能力低,资源极其匮乏,对蛇足石杉的快速繁殖技术进行探讨。方法取活体茎尖,依次用多种消毒剂进行表面消毒,再用孔雀石绿及原子吸收光谱(AAS)联合杀灭内生真菌,得到无菌苗接种到1/2MS+25 g/L蔗糖的培养基中,无菌苗在二叉分枝后的新枝长到4~5 cm时剪下进行继代培养。结果无菌苗分别在4次二叉分枝生长继代培养后,得到了数量不等的侧芽,侧芽的发生位置位于茎段中部或上部近茎尖处,侧芽的结构与芽孢的结构明显不同;不同植株培养获得的侧芽数不同,最多的植株可产生14个侧芽,一个侧芽发生点可产生一个或多个侧芽;去除茎尖顶端优势后,茎段中部的侧芽生长速度加快;不同高度的侧芽拨离母株后,成活率差别大,5 mm以上的侧芽接种到1/2 MS+30 g/L蔗糖的培养基上,存活率可达90%,120 d后生长为2~3 cm的幼苗,低于5 mm的侧芽,拨离接种后死亡率较高。结论蛇足石杉在离体培养中,可打破自然生长状态下叶腋内只产生孢子囊而不产生侧芽的生长方式,可形成多个增殖侧芽,侧芽剥离后极易产生不定根,为蛇足石杉的试管苗增殖方式提供重要参考。
Objective Huperzia serrata has low natural reproductive capacity and extremely scarce resources. The rapid propagation technology of Huperzia serrata is discussed. Methods The tips of the in vivo shoots were disinfected with several kinds of disinfectants, then the endophytic fungi were killed by combining malachite green and atomic absorption spectrometry (AAS). The aseptically inoculated sterile medium was inoculated into 1 / 2MS + 25 g / L sucrose Of the medium, sterile seedlings in the new branch after the bifurcation to 4 ~ 5 cm cut off subculture. Results Aseptic shoots were obtained after 4 generations of subculture, respectively. The number of lateral buds was located in the middle or upper part of shoots. The structure of lateral buds was obviously different from that of spores ; The number of lateral buds obtained by different plants is different, the most plants can produce 14 lateral buds, one bud can produce one or more lateral buds; the removal of top tips of stem tips, the lateral buds grow faster in the middle of stems; different height After the lateral buds were removed from the parental plants, the survival rate varied greatly. The lateral buds of more than 5 mm were inoculated into 1/2 MS + 30 g / L sucrose medium, the survival rate was 90%, and the growth was 2 ~ 3 cm after 120 days Of young shoots, lateral buds below 5 mm, higher mortality rates after dialysis vaccination. Conclusion In the in vitro culture, the genus Hippeastrum can break the growth mode in which only the sporangia are produced in the axilla of the leaves without producing lateral buds, and multiple lateral buds can be formed. After the lateral buds are stripped, adventitious roots are easily produced, Test tube proliferation provides an important reference.