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【目的】尝试使用茎尖分生组织培养技术生产UP 01235和UP 0097 2个甘蔗品种的脱甘蔗黄叶病毒(SCYLV)脱毒组培苗。【方法】以大田染病和健康甘蔗植株的不同大小茎尖分生组织为外植体,经培养获得组培幼苗。采用1对甘蔗黄叶病毒特异性引物(SCYLV-615F:5′-ATGAATACGGGCGCTAACCGYYCAC-3′和 SCYLV-615R:5′-GT-GTTGGGGRAGCGTCGCYTACC-3′)进行RT-PCR分析,在6个月内每隔1个月检查1次组培苗是否存在病毒。【结果】当以1.5 mm茎尖进行培养时,获得的2个甘蔗品种的组培苗在温室种植,在生长期间未发现存在黄叶病毒,说明黄叶病毒已被消除;而以超过1.5 mm的茎尖为外植体时,PCR分析表明结果,所获得的组培苗存在黄叶病毒。【结论】对以1.0～1.5 mm茎尖培养获得的200多株组培苗进行温室繁殖,在生长期间均未发现黄叶病毒。
【Objective】 The purpose of this experiment was to produce virus-free tissue culture seedlings of Sugarcane Sugarcane Leaf Virus (SCYLV) from two sugarcane varieties UP 01235 and UP 0097 using shoot apical meristem culture. 【Method】 Tissue-derived shoots of Diantian-infected and healthy sugarcane plants of different sizes were used as explants. RT-PCR analysis was performed using 1 pair of sugarcane yellow leaf virus-specific primers (SCYLV-615F: 5’-ATGAATACGGGCGCTAACCGYYCAC-3 ’and SCYLV-615R: 5’-GT- GTTGGGGRAGCGTCGCYTACC-3’) at intervals of 6 months 1 month inspection tissue culture seedling virus. 【Result】 When 1.5 mm shoot tip was used for culture, the tissue culture seedlings of two sugarcane varieties were planted in the greenhouse. No yellow leaf virus was found during the growth period, which indicated that the yellow leaf virus had been eliminated. However, When the shoot tips were explants, PCR analysis showed that the obtained tissue culture seedlings contained the yellow leaf virus. 【Conclusion】 More than 200 plantlets were obtained from the tips of 1.0-1.5 mm shoots in the greenhouse and no yellow leaf virus was found during the growth.