新疆雪莲叶片高效再生植株体系的建立

Establishment of High Frequency Plant Regeneration System From the Leaf of Saussurea involucrate

以新疆雪莲快繁无菌苗为试材，研究了新疆雪莲高效植株再生的条件。结果表明：长势健壮的无菌苗叶片切段可以通过2种途径高效再生不定芽；直接器官再生途径：长3～51mm的叶切段在MS＋NAA0．5mg／L＋6-BA2．0mg／L的培养基上先后暗诱导3周和光诱导4周，分化率达（73．3±3．8）%，平均出芽量（5．0±0．3）个芽／块；间接器官再生途径：叶切段在MS＋NAA0．2mg／L＋TDZ0．5mg／L培养基上暗诱导3周，愈伤组织诱导率1000，愈伤组织在MS＋NAA0．5mg／L＋6-BA2．0mg／L的培养基上光诱导4周，分化率达（88．9±5．9）9／6，平均（5．5±0．4）个芽／块；不定芽在MS＋NAA0．05mg／L＋6-BA0．3mg／L培养基中壮苗和增殖培养后，在MS＋IBA0．5mg／L＋0．1％活性碳的培养基中生根率达100％；2种植株再生途径均可用于新疆雪莲的诱变育种和转基因育种研究。

Taking proliferated sterile seedlings of Saussurea involucrate as plant material, appropriate conditions for high frequency plant regeneration of Saussurea involucrate were investigated. The results showed that robust leaves of the seedlings could be induced to regenerate the plant in two ways. Direct regeneration way： leaf fragments in length of 3- 5 mrn were inoculated on the medium of MS＋NAA 0. 5 mg/L＋6-BA 2. 0 mg/L. The differentiation rate could reach （73. 3±3. 8）% after being cultured for 3 weeks in darkness and 4 weeks under the light. The fragments could produce healthy adventitious shoot clusters with an average number of （5.0±0. 3） shoots/explant. Indirect regeneration way. calli were induced from leaf fragments inoculated on the medium of MS＋NAA 0. 2 mg/L＋TDZ 0. 5 mg/L for 3 weeks in darkness,and then were cultured on medium of MS＋ NAA 0. 5 mg/L＋6-BA 2. 0 mg/L for 4 weeks under the light. （88. 9±5.9）% of calli could produce adventitious shoot with an average number of （5. 5±0. 4） shoots/callus. After growing on the medium of MS＋NAA 0. 05 mg/L＋6-BA 0. 3 mg/L for 3 to 4 weeks,the shoots proliferated and became stronger. Thus,almost 1.00% of the shoots could root well on the medium of MS＋IBA 0. 5 mg/L＋0. 1%active carbon. The two protocols could be used for genetic transformation research of S. involucrate.