广丰药薯试管苗离体快繁技术的研究

Study on Rapid Micropropagation in Vitro Technique of Guangfeng Medicinal Yam( Dioscorea opposita) Plantlets

目的：对广丰药薯离体快繁技术进行研究,为广丰药薯试管苗的脱毒和种质保存提供方法学参考。方法：采用植物组织培养和单因子试验的方法,研究广丰药薯组织培养的基本程序,并利用流式细胞术检测广丰药薯试管苗与盆栽苗的DNA含量。结果：广丰药薯带芽茎段消毒的较佳程序是70%酒精灭菌20~30 s后无菌水冲洗3次,再用0.1%氯化汞灭菌10~12 min后无菌水冲洗3次;广丰药薯消毒的较佳外植体是稍木质化较成熟的带芽茎段;广丰药薯带芽茎段芽增殖较佳的培养基为MS＋6-BA 2.0 mg/L＋NAA 0.1 mg/L;广丰药薯带芽茎段生根较佳的培养基为MS＋NAA 0.5 mg/L;广丰药薯试管苗较佳的培养方式为液体培养;广丰药薯试管苗移栽的较佳基质为稻田粘土与细砂（1∶2）混匀物或珍珠岩与蛭石（1∶2）混匀物;广丰药薯试管苗的DNA含量与盆栽苗相比无显著性差异。结论：该研究建立了广丰药薯带芽茎段的离体快繁技术体系,其试管苗的流式细胞术检测结果也表明了广丰药薯试管苗的遗传稳定性,为广丰药薯试管苗的规模化生产提供了技术基础和理论依据。

Objective： In order to provide methodology reference for virus-free and germplasm conservation of Guangfeng medicinal yam（ Dioscorea opposita） plantlets,rapid micropropagation in vitro technique of Guangfeng medicinal yam plantlets was studied. Methods： Using the method of plant tissue culture,single factor test and flow cytometry,the basic procedure of Guangfeng medicinal yam tissue culture was established and the DNA content of Guangfeng medicinal yam plantlets and its potted seedlings was detected. Results：The best disinfection procedure of stems with a bud of Guangfeng medicinal yam was washed with sterile water for three times after sterilized with 70% alcohol for 20 ~ 30 s and then washed with sterile water for three times again after sterilized with 0. 1% mercuric chloride for 10 ~ 12 min; The best explants of stems with a bud of Guangfeng medicinal yam was slightly woody and more mature stems with a bud; The best proliferation culture medium of stems with a bud of Guangfeng medicinal yam was MS ＋ 6-BA 2. 0 mg / L ＋ NAA 0. 1mg / L; The best rooting culture medium of stems with a bud of Guangfeng medicinal yam was MS ＋ NAA 0. 5 mg / L; The best culture method of Guangfeng medicinal yam plantlets was liquid culture; The best transplanting matrix of Guangfeng medicinal yam plantlets was the mixture of paddy clay and fine sand（ 1∶ 2） or the mixture of perlite and vermiculite（ 1∶ 2）; The DNA content between Guangfeng medicinal yam plantlets and its potted seedlings had no significant difference. Conclusion： A fast and efficient micropropagation in vitro technological system of stems with a bud of Guangfeng medicinal yam is established,and the flow cytometry detect results also show the genetic stability of Guangfeng medicinal yam plantlets,whose results provide the technical and theoretical basis for the large-scale production of Guangfeng medicinal yam plantlets.