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吕经娟, 李淑斌, 周宁宁, 蹇洪英, 王其刚, 张颢, 唐开学. ‘光叶蔷薇’器官间接再生体系的建立及GUS基因转化的初步研究[J]. 植物科学学报, 2014, 32(2): 139-147. DOI: 10.3724/SP.J.1142.2014.20139
引用本文: 吕经娟, 李淑斌, 周宁宁, 蹇洪英, 王其刚, 张颢, 唐开学. ‘光叶蔷薇’器官间接再生体系的建立及GUS基因转化的初步研究[J]. 植物科学学报, 2014, 32(2): 139-147. DOI: 10.3724/SP.J.1142.2014.20139
LÜ Jing-Juan, LI Shu-Bin, ZHOU Ning-Ning, JIAN Hong-Ying, WANG Qi-Gang, ZHANG Hao, TANG Kai-Xue. Callus Induction and Plant Regeneration of Rosa wichuraiana ‘Basye’s thornless’[J]. Plant Science Journal, 2014, 32(2): 139-147. DOI: 10.3724/SP.J.1142.2014.20139
Citation: LÜ Jing-Juan, LI Shu-Bin, ZHOU Ning-Ning, JIAN Hong-Ying, WANG Qi-Gang, ZHANG Hao, TANG Kai-Xue. Callus Induction and Plant Regeneration of Rosa wichuraiana ‘Basye’s thornless’[J]. Plant Science Journal, 2014, 32(2): 139-147. DOI: 10.3724/SP.J.1142.2014.20139

‘光叶蔷薇’器官间接再生体系的建立及GUS基因转化的初步研究

Callus Induction and Plant Regeneration of Rosa wichuraiana ‘Basye’s thornless’

  • 摘要: 以‘光叶蔷薇’(Rosa wichuriana ‘Basye’s thornless’)无菌苗的顶生幼嫩小叶为外植体,探讨了其愈伤组织诱导及植株再生的方法。结果表明,高浓度的生长素NAA能诱导外植体产生愈伤组织;由NAA诱导的愈伤组织在附加TDZ的MS培养基上,先暗培养再进行光照培养可直接分化出不定芽。诱导愈伤组织的最佳NAA浓度是7.0 mg/L、暗培养时间为10 d,而最佳分化培养基是MS + 5.0 mg/L TDZ + 30 g/L葡萄糖 + 2.5 g/L GEL,分化率达18.34%。以诱导产生的愈伤组织为侵染受体,初步建立了‘光叶蔷薇’GUS基因转化体系。农杆菌菌液浓度OD600值为0.5、侵染30 min、共培养2 d、乙酰丁香酮的浓度为50 μmol/L是'光叶蔷薇’愈伤组织转基因的最优条件。

     

    Abstract: Methods of callus induction and plant regeneration of Rosa wichuraiana ‘Basye's thornless’ were investigated using in vitro grown young leaflets as explants. Results showed that calli were induced on MS with a high concentration of auxin (NAA), with calli exhibiting direct differentiation of adventitious buds after induction on MS medium with TDZ under dark and then light conditions. The optimal concentration of auxin (NAA) for calli induction of R. wichuraiana was 7.0 mg/L, the optimal induction period in the dark for adventitious bud differentiation was 10 d and the optimal differentiation medium was MS + 5.0 mg/L TDZ + 30 g/L glucose + 2.5 g/L GEL (pH 5.8-6.0). The highest rate of adventitious bud induction was 18.34%. We used the calli obtained by induction as transformation receptors to research GUS gene conversion. We also conducted a preliminary study on the Agrobacterium tumefaciens-mediated transformation protocol for R. wichuraiana. Results showed that the optimal conditions for transformation of R. wichuraiana was a bacterial concentration (OD600) of 0.5, infection time of 30 min, co-cultivation time of 2 d, and acetosyringone concentration of 50 μmol/L.

     

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