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利用基因枪法将来源于大麦(Hordeum vulgare L.)的抗旱耐盐基因lea3导入铁皮石斛(Dendrobium candidum Wall.ex Lindl)的类原球茎(PLBs)中,经膦丝菌素类除草剂PPT筛选2次获得抗性PLBs,于无选择压下分化获得不定芽,再经PPT筛选和生根壮苗培养获得转化植株.对转化植株进行除草剂PPT叶片涂抹检测和lea3基因的PCR检测,表明lea3基因已整合到6个株系7株铁皮石斛转化植株基因组中,转化频率为1.05%.与对照相比,获得的转基因植株的耐盐胁迫能力明显增强,表明遗传转化lea3基因可用于石斛抗旱耐盐新品种的选育.
Drought-resistant and salt-tolerant gene lea3 derived from Hordeum vulgare L. was introduced into PLBs by biolistic method and screened by PPT of phosphinothricin herbicide 2 Resistant PLBs were obtained and differentiated to obtain adventitious buds without selection and then transformed into plants by PPT screening and rooting and seedling cultivation.The herbicide PPT leaf smear detection and lea3 gene PCR analysis of transformed plants showed that lea3 gene The transformation frequency was 1.05% in the transformed plants of seven strains of Dendrobium candidum in six lines.The tolerance of salt-tolerant plants to transgenic plants was significantly enhanced compared with the control, indicating that the genetic transformation of lea3 gene can be used for drought resistance and salt tolerance of Dendrobium Breeding of varieties.