翠竹鞭芽组培快繁高效再生技术体系的构建

Establishing An Efficient Regeneration System for Tissue Culture in Pleioblastus pygmaeus with Rhizome Lateral Buds

2015年5月20日,将采于江苏省南京市南京林业大学白马基地竹种园的翠竹(Pleioblastus pygmaeus(Miq.)E.G.Camus)种子播种于培养基质体积比为V(泥炭)∶V(黄棕壤)∶V(珍珠岩)=12∶12∶1混合基质的花盆中,定期进行管理。2019年,将翠竹实生苗包括鞭根系统及基质整体取出,选取翠竹竹鞭上带有未萌动和刚萌动鞭芽的鞭段(约1.5 cm)作为外植体,构建翠竹离体培养再生技术体系。以污染率、成活率、增殖系数、生根率为评价指标,分析不同消毒方法的效果、不同生长调节剂组合对翠竹继代增殖和诱导生根的影响。结果表明:“体积分数为70%的酒精处理60 s+体积分数为2.5%的NaClO溶液处理15 min”对鞭芽的消毒效果最好,成活率可达(94.62±0.72)%。翠竹鞭芽继代增殖培养的最佳培养基为“MS培养基+质量浓度为5 mg·L^(-1)的6-苄基氨基嘌呤(6-BA)+质量浓度为0.3 mg·L^(-1)的萘乙酸(NAA)”,增殖系数为7.24±0.23。生根培养的最佳培养基为“0.5倍的MS培养基+质量浓度为1 mg·L^(-1)的6-苄基氨基嘌呤+质量浓度为1.5 mg·L^(-1)的萘乙酸+质量浓度为0.5 mg·L^(-1)的吲哚丁酸(IBA)”,生根率可达90%,平均生根数增长速度最快。将瓶苗移栽到体积比为V(壤土)∶V(蛭石)=1∶1的基质中,成活效果最高,为100%。

The seeds of Pleioblastus pygmaeus(Miq.)E.G.Camus collected in Baima bamboo base of Nanjing Forestry University in Jiangsu Province were sown in a flowe rpot with a mixed substrate volume ratio of V(peat)∶V(yellow brown soil)∶V(perlite)of 12∶12∶1 on May 20,2015,and the seedlings were managed regularly.In 2019,the seedlings including the rhizome root system and the substrate were taken out and then the soil was washed,the rhizome segments with non-germinating and germinating buds(about 1.5-cm long)were selected as the explants to construct in vitro regeneration system.Taking the pollution rate,survival rate,proliferation coefficient and rooting rate as evaluation indexes,the effects of different disinfection methods and different combinations of growth regulators on the subculture proliferation and induced rooting were analyzed.Treatment with“70%alcohol for 60 s+2.5%NaClO solution for 15 min”has the best disinfection effect on the rhizome bud,and the survival rate could reach 94.62±0.72%.The best medium for subculture and proliferation of rhizome buds is“MS medium+6-Benzylaminopurine(6-BA)with a mass concentration of 5 mg·L^(-1)+NAA with a mass concentration of 0.3 mg·L^(-1)”,and the proliferation coefficient is 7.24±0.23.The best medium for rooting culture is“0.5 times MS medium+6-Benzylaminopurine with a mass concentration of 1 mg·L^(-1)+NAA with a mass concentration of 1.5 mg·L^(-1)+indole butyric acid(IBA)with a mass concentration of 0.5 mg·L^(-1)”.The rooting rate could reach 90%,and the average rooting number increases the fastest.The seedlings were transplanted into a matrix with a volume ratio of V(soil)∶V(vermiculite)=1∶1,and the survival effect was the highest,which was 100%.