蝴蝶兰茎尖培养脱病毒技术初步研究

摘要

The effect of culture medium and explant on the tip-shoot tissue culture for eradicating orchid virus was studied. The concentrations of inorganic salt, nitrogen, potassium, and plant growth regulator in culture medium were tested and optimized. The results showed that 1/5 inorganic salt quantity of KN medium improved from the MS medium is suitable to tip-shoot culture of Phalaenopsis. Within a certain concentration range, increasing the proportion of nitrate nitrogen in total nitrogen content and the potassium concentration proportion in total inorganic salt of medium contribute to improve the survival of tip-shoot culture. The plant growth regulator combination of NAA 0.5 mg·L-1+6-BA 3.5 mg·L-1 is suitable to the Protocorm Like Bodies (PLBs) inducted from the Phalaenopsis tip-shoot. Big drawn size of tip-shoot organization is conductive to the survival of tip-shoot culture but detrimental to virus-eradicting effect. The orchid virus ORSV is more difficult to deal with than the orchid CymMV. Drawn size smaller than 0.4 mm, attaching leaf primordium no more than one is necessary for thorough eradication of orchid virus by the way of tip-shoot culture.%　　对培养基的无机盐浓度、钾浓度比例、氮素型态、植物生长调节剂进行了试验优化，研究了培养基和外植体取材对蝴蝶兰茎尖生长点培养脱病毒的影响。结果表明：经MS培养基改良的KN培养基，其1/5大量元素的无机盐浓度适合蝴蝶兰茎尖生长点的培养。一定浓度范围内，提高氮源中硝酸态氮的比例，或提高培养基中钾的浓度比例，均有助于蝴蝶兰茎尖生长点培养的存活。植物生长调节剂NAA 0.5 mg·L-1+6-BA 3.5 mg·L-1的组合适合蝴蝶兰茎尖生长点培养的存活和类原球茎(Protocorm-Like Body,PLBs)的诱导。茎尖取材大利于培养存活，但是不利于脱病毒效果。齿舌兰轮斑病毒(简称ORSV)的茎尖生长点脱病毒难度较建兰花叶病毒(简称CymMV)的脱病毒难度大。蝴蝶兰茎尖培养脱病毒取材外植体须小于0.4 mm，附带叶原基不能多于1个。