黄果百香果茎段愈伤组织培养脱毒及脱毒苗快繁体系建立

Establishment of Virus-free Callus Culture and Rapid Propagation System of Virus-free Seedlings from Stem Segments of Yellow Passion Fruit

以福建百香果3号植株的新枝为材料,对其茎切面愈伤组织诱导,不定芽诱导分化成苗,采用酶联免疫吸附法检测百香果斑驳病毒(PaMV)、夜来香花叶病毒(TeMV)、东亚西番莲病毒(EAPV)、西番莲潜隐病毒(PLV)、黄瓜花叶病毒(CMV)等5种病毒。挑选病毒阴性的继代苗扩繁培养,优化不同培养阶段中适宜的激素种类、浓度配比组合,为建立百香果脱毒离体组培技术体系提供参考依据。结果表明:在黄果百香果愈伤组织的诱导中,以MS培养基+6-BA 1.5 mg/L+NAA 0.2 mg/L+蔗糖20 g/L+琼脂7 g/L的组合诱导效果最好,其诱导出愈率为83.33%;在愈伤组织诱导分化不定芽中,以MS培养基+6-BA 0.8 mg/L+NAA 0.1 mg/L+土豆泥200 g/L+蔗糖20 g/L组合的芽诱导效果最好,其不定芽诱导率为46.70%;在不定芽诱导成苗中,以MS培养基+IBA 1.0 mg/L+NAA 0.2 mg/L的激素浓度组合的诱导生根效果最好,其生根率达93.33%。病毒检测试验发现:20份样品中有9份均不含PaMV、TeMV、EAPV、PLV和CMV,脱毒率为45%;脱毒苗快繁培养中,以MS培养基+6-BA 0.5 mg/L+IBA 0.2 mg/L+蔗糖30 g/L+琼脂粉5 g/L组合的效果最好,表现为生根快、茎粗壮,幼苗长势好。

The robust stem segments of yellow passion fruit“Fujian passion fruit No.3”were used as materials to induce callus,induce adventitious buds to differentiate into seedlings,and detect passion fruit mottle virus(PAMV),evening Jasmine mosaic virus(temv),East Asia passion fruit virus(eapv),passion fruit latent virus(PLV)and cucumber mosaic virus(CMV)by enzyme-linked immunosorbent assay.The virus negative subculture seedlings were selected for propagation and culture,and the appropriate hormone types and concentration combinations in different culture stages were optimized.The results showed that the combination of MS+6-BA 1.5 mg/L+NAA 0.2 mg/L+sucrose 20 g/L+agar 7 g/L had the best induction effect,and the induction rate was 83.33%.Among the callus induced adventitious buds,the combination of MS+6-BA 0.8 mg/L+NAA 0.2 mg/L+mashed potato 200 g/L+sucrose 20 g/L had the best effect,and the adventitious bud induction rate was 46.7%.In the adventitious bud induction,the combination of MS+IBA 1.0 mg/L+NAA 0.2 mg/L hormone concentration had the best rooting effect,and the rooting rate was 93.33%.Virus detection experiments showed that 9 of the 20 samples did not contain CMV,PaMV,TEeMV,EAPV and PLV,and the virus-free rate was 45.0%.In the rapid propagation culture of virus-free seedlings,the combination of MS+6-BA 0.5 mg/L+IBA 0.2 mg/L+sucrose 30 g/L+agar powder 5 g/L had the best effect,showing fast rooting,thick stems and good seedling growth.