PJB-2024-105
Optimization of hairy root transformation system for gene functional studies in poplar (Populus species)
Mei Han, Junhu Feng, Shuyue Sun, Yujia Si, Xiaoning Liu, Mingyue Xu and Tao Su
DOI:10.30848/PJB2025-4(5)
Abstract
Poplars (Populus spp.) are one of the most widespread ligneous plants in the world, which have a variety of applications ranging from landscape, agriculture, and industry to research. Although Agrobacterium tumefaciens-mediated gene transformation method has been widely utilized for poplar genetic manipulation, many defective technique defects such as the time-consuming procedures, and the occurrence of chimeric transformant have emerged. To address these problems, a simple, fast, straightforward and effective gene transformation method by using Agrobacterium rhizogenes to induce hairy roots in poplars was developed, and regenerated-transgenic poplar plantlets were obtained within a period of two to three months. The optimal procedure comprised the usage of poplar apex buds as explants, infection of A. rhizogenes by directly pricking colonies with the cuttings on the agar plates, and improving hairy-root initiation by application of indole-3-butyric acid (IBA) in combination with hairy-root morphological identification. Moreover, the GFP marker gene was applied to track record the expression level of the introduced genes using a portable fluorescence lamp. Further quantitative real-time PCR (qRT-PCR) and the enzyme activity assay showed that in comparison with the non-transformed control, the transcript abundance and enzyme activity of our target gene (i.e., aspartate aminotransferase) were significantly elevated in the transformed plant lines regenerated from the hairy roots, confirming that the current A. rhizogenes-mediated poplar transformation method is effective and applicable for poplar genetic manipulation and functional genomic study
To Cite this article:
Han, M., J. Feng, S. Sun, Y. Si, X. Liu, M. Xu and T. Su. 2025. Optimization of hairy root transformation system for gene functional studies in poplar (Populus species). Pak. J. Bot., 57(4): DOI: http://dx.doi.org/10.30848/PJB2025-4(5)
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