姜荷花‘红色印象’工厂化组培育苗技术研究

Industrial Tissue Culture Speed Propagation of Curcuma alismatifolia‘Majeo Impress Red’

以姜荷花‘红色印象’植株的多个部位作为外植体,进行丛生芽诱导、增殖扩繁、生根壮苗以及出瓶移栽育苗的研究,完善姜荷花工厂化组培快繁流程,为其规模化生产提供参考。以姜荷花球茎、储藏根、球茎小芽、侧芽、幼嫩花芽、幼嫩叶片6个部位为材料,通过添加不同浓度6-BA和NAA的MS培养基,对丛生芽诱导培养、丛生芽增殖培养、生根壮苗以及移栽育苗等步骤进行优化。姜荷花‘红色印象’最佳外植体为球茎小芽和侧芽,诱导培养基为MS+6-BA(3 mg/L或5 mg/L)+NAA 0.2 mg/L+蔗糖30 g/L+卡拉胶7 g/L,最佳丛生芽增殖培养基为MS+6-BA 3 mg/L+NAA 0.1 mg/L+蔗糖30 g/L+卡拉胶7 g/L,生根壮苗培养基选用1/2 MS+NAA 0.1 mg/L+蔗糖20 g/L+卡拉胶7 g/L,经过炼苗后出瓶移栽,组培苗种植基质选用细泥炭和细椰糠1∶1混合基质最佳,出瓶移栽成活率达95%以上,表型稳定。利用EST-SSR标记对母株和随机选择的组培苗进行扩增分析,证实DNA水平上无明显变异。通过对姜荷花‘红色印象’外植体和培养基的筛选,实现姜荷花组培快繁流程优化,建立高效完整的姜荷花工厂化组培育苗体系。

In order to improve the technological process of industrial tissue culture speed propagation of Curcuma alis-matifolia‘Majeo Impress Red’,and provide a reference for the large-scale production,we used different organs as the explants to study the bud multiplication,proliferation,rooting culture and transplanting.Six different explants including corm,storage root,corm shoot,lateral bud,flower bud and tender leaves were employed in this study.Based on the MS medium with different concentration of 6-BA and NAA,the process of tissue culture speed propagation was optimized.The best explants were corm shoot and lateral bud.The best medium for induction culture was MS+6-BA 3 mg/L+NAA 0.1 mg/L+sucrose 30 g/L+carrageenans 7 g/L.The best medium for proliferation was MS+6-BA 3 mg/L+NAA 0.1 mg/L+sucrose 30 g/L+carrageenans 7 g/L.The best medium for rooting was 1/2 MS+NAA 0.1 mg/L+sucrose 20 g/L+carrageenans 7 g/L.When the seedlings were pluged in tray and transplanted in flowerpot,the optimum medium was tiny peat∶tiny coco coir=1∶1 and the survival rate was over 95%.Eight EST-SSR markers were employed to amplified the maternal parent and 20 randomly selected tissue culture seedlings,which supported the obsence of DNA-level variations.We screened the most appropriate explants and culture medium for C.alismatifolia‘Majeo Impress Red’,optimized the technological process and set up a technical system of industrial tissue culture speed propagation.