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以感染菊花B病毒(CVB)的菊花品种‘寒小白’为试材,分别切取长度小于0.3 mm、0.3～0.7 mm和0.7～1.0 mm的茎尖接种于MS+2.0 mg·L-16-BA+0.5 mg·L-1NAA的培养基中,以及第1次切取0.7～1.0 mm菊花茎尖接种于MS培养基中,培养后再切取0.3～0.7 mm的菊花茎尖接种于MS+2.0 mg·L-16-BA+0.5 mg·L-1NAA培养基中(二次茎尖培养法),结合RT-PCR方法鉴定脱毒效果,研究茎尖长度及培养方式对茎尖诱导分化成活率和脱毒效果的影响。结果表明:茎尖大小与茎尖诱导成活率成正比,与脱毒效果成反比,即茎尖长度小于0.3 mm时,成活率最低,仅为6%,而脱毒率最高,可达66.67%;当茎尖长度为0.7～1.0 mm时,茎尖成活率最高,为90%,脱毒率最低,仅为6.67%。在一次茎尖培养法的基础上,运用二次茎尖培养法对CVB进行脱毒培养,既保证了较高的成活率(60%),又保证了较高的脱毒率(43.33%)。
In order to infect Chrysanthemum breviflora chinensis var. Chinensis var. Chinensis var. Chinensis var. Chinensis CVB was inoculated into MS + 2.0 mg · L-16- BA + 0.5 mg · L-1 NAA, and the 1st cut chrysanthemum shoot tip of 0.7-1.0 mm was inoculated into MS medium. After inoculation, 0.3-0.7 mm chrysanthemum shoots were inoculated into MS + 2.0 mg · L-16-BA + 0.5 mg · L-1NAA medium (secondary shoot tip culture method), combined with RT-PCR method to identify the detoxification effect, to study the shoot tip length and culture methods on shoot tip differentiation and survival rate and Effects of detoxification. The results showed that the size of shoot tip was proportional to the survival rate induced by shoot tip, which was inversely proportional to the detoxifying effect. When the shoot tip length was less than 0.3 mm, the survival rate was the lowest, only 6% while the highest detoxification rate was 66.67% When the shoot tip length was 0.7-1.0 mm, the shoot tip had the highest survival rate of 90% with the lowest detoxification rate of 6.67%. On the basis of a shoot tip culture method, the secondary shoot tip culture method was used to detoxify the CVB, which not only guaranteed a higher survival rate (60%), but also ensured a higher detoxification rate (43.33%). .