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Efficient Agrobacterium-Mediated Transformation of the Commercial Hybrid Poplar Populus Alba × Populus glandulosa Uyeki

机译:农杆菌介导的商业化杂交杨白杨×灰杨Uyeki的高效转化

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摘要

Transgenic technology is a powerful tool for gene functional characterization, and poplar is a model system for genetic transformation of perennial woody plants. However, the poplar genetic transformation system is limited to a number of model genotypes. Herein, we developed a transformation system based on efficient Agrobacterium-mediated transformation for the hybrid poplar Populus Alba × Populus glandulosa Uyeki, which is a fast-growing poplar species that is suitably grown in the northern part of China. Importantly, we optimized many independent factors and showed that the transformation efficiency was improved significantly using juvenile leaf explants. Explants were infected by an Agrobacterium suspension with the OD600 = 0.6 for 15 min and then co-cultured in dark conditions for 3 days. Using the improved transformation system, we obtained the transgenic poplar with overexpression of β-glucuronidase (GUS) via direct organogenesis without callus induction. Furthermore, we analyzed the GUS gene in the transgenic poplars using PCR, qRT-PCR, and GUS staining. These analyses revealed that the GUS gene was efficiently transformed, and it exhibited various expression levels. Taken together, these results represent a simple, fast, and efficient transformation system of hybrid poplar plants. Our findings may facilitate future studies of gene functions in perennial woody plants and tree breeding via transgenic technology assisted design.
机译:转基因技术是进行基因功能鉴定的有力工具,而杨树是多年生木本植物遗传转化的模型系统。但是,杨树遗传转化系统仅限于多种模型基因型。在本文中,我们开发了一种基于高效农杆菌介导的转化系统,用于杂种杨(Populus Alba)×Populus glandulosa Uyeki(一种快速生长的杨树种,适合在中国北部种植)。重要的是,我们优化了许多独立因素,并表明使用幼叶外植体可显着提高转化效率。用OD600 = 0.6的农杆菌悬液将外植体感染15分钟,然后在黑暗条件下共培养3天。使用改良的转化系统,我们通过不经愈伤组织诱导的直接器官发生获得了具有过表达β-葡萄糖醛酸苷酶(GUS)的转基因杨树。此外,我们使用PCR,qRT-PCR和GUS染色分析了转基因杨树中的GUS基因。这些分析表明,GUS基因被有效地转化,并且表现出各种表达水平。综上所述,这些结果代表了杂交杨树植物的简单,快速和有效的转化系统。我们的发现可能有助于通过转基因技术辅助设计对多年生木本植物和树木育种中基因功能的未来研究。

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