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在用农杆菌介导法转化甘蓝型油菜的组织培养过程中,常会伴生着试管苗玻璃化现象的发生,此现象的发生会降低转基因的频率.本试验结果表明,影响玻璃化发生的因素较多,包括激素体积质量分数、蔗糖体积质量分数、琼脂体积质量分数和接种密度等.其中,激素6-BA和ZT体积质量分数对诱芽率和玻璃化率都有显著影响,随着体积质量分数的升高,玻璃化率升高,以6-BA体积质量分数为1mg/L,ZT体积质量分数为2mg/L时结果较为理想.蔗糖体积质量分数和琼脂体积质量分数对玻璃化率也有显著影响,当二者体积质量分数升高,玻璃化率降低,同时诱芽率也降低.综合分析,当蔗糖体积质量分数为50g/L,琼脂体积质量分数为10g/L时最适宜.接种密度对玻璃化的发生有显著影响,密度越高,玻璃化倾向越严重,而密度对诱芽率影响不明显.试验结果分析发现,接种密度为20~30个/皿最适宜.
In Agrobacterium tumefaciens-mediated transformation of Brassica napus tissue culture, often accompanied by the occurrence of plantlet vitrification, the occurrence of this phenomenon will reduce the frequency of the transgene.This test results show that the impact of vitrification than the factors Including hormonal volume fraction, sucrose volume fraction, agar volume fraction and inoculation density, etc. Among them, hormone 6-BA and ZT volume fraction had significant effects on the rate of budding and vitrification, The results showed that when the mass fraction of 6-BA was 1mg / L and the volume fraction of ZT was 2mg / L, the results were satisfactory.The volume fraction of sucrose and the volume fraction of agar also had the same effect on the vitrification rate The results showed that when the mass fraction of sucrose was 50g / L and the volume fraction of agar was 10g / L, the optimum conditions were as follows: Density has a significant effect on the occurrence of vitrification, the higher the density, the more serious the vitrification tendency, while the density has no obvious effect on the rate of seed germination.According to the test results, it is found that the optimal density is 20 ~ 30 per dish.