Agrobacterium rhizogenes-mediated hairy roots transformation as a tool for exploring aluminum-responsive genes function
Agrobacterium rhizogenes-mediated hairy roots transformation as a tool for exploring aluminum-responsive genes function
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DOI：
10.4155/fsoa-2018-0065
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发表时间：
2019-03-01
影响因子：
2.5
通讯作者：
Koyama, Hiroyuki
中科院分区：
文献类型：
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作者：
Daspute, Abhijit A.;Xian Yunxuan;Koyama, Hiroyuki
目的：为毛状根基因功能的研究提供一种新的方法。方法：以拟南芥和烟草(野生型和突变型)为寄主，进行发根农杆菌转化。结果：烟草和拟南芥的毛状根形成率分别为53%~98%和53%~66%。毛状根和完整根在盐和铝胁迫下表现出相似的基因表达模式。基因组聚合酶链式反应和荧光图像显示，在不使用任何抗生素选择的情况下，T-DNA的共整合率很高(>；80%)，细胞转化均匀。农杆菌侵染后1个月内即可完成毛状根的整个生长过程。结论：NtSTOP1-KD和Atstop1可以作为毛状根转化的宿主，评价铝敏感的同源基因的功能。在本研究中，我们使用发根农杆菌介导的毛状根作为农杆菌转化的替代方法。我们在烟草和拟南芥中开发了一种简单、有效和可重复性的毛状根方案。将发育的毛状根与完整的毛状根进行比较，以研究其对铝、盐胁迫的基因响应和基因的细胞定位。利用NtSTOP1-KD和Atstop1作为毛状根转化的宿主可以评价铝敏感的同源基因的功能。
Aim: To develop a useful alternative approach to evaluate the gene function in hairy roots. Methods:Arabidopsis and tobacco (wild-type or mutant) were a host for Agrobacterium rhizogenes transformation. Results: The hairy roots formation efficiency ranged from 53 to 98% in tobacco and 53 to 66% in Arabidopsis. Hairy and intact roots showed similar gene expression pattern in response to salt and aluminum stress. Genomic polymerase chain reaction and fluorescent images showed high rate (>80%) of co-integration of T-DNAs and uniform cell transformation without use of any antibiotic selection. Whole processes of hairy roots were completed within 1 month after the infection of Agrobacterium. Conclusion: Aluminum-responsive orthologous gene function could be evaluated by NtSTOP1-KD and Atstop1 as a host for hairy roots transformation.Lay abstract: Hairy roots have been used for various purposes over the last several years. In the present study we used Agrobacterium rhizogenes-mediated hairy roots as an alternative approach for Agrobacterium tumefaciens transformation. We developed a simple, effective and reproducible hairy root protocol in tobacco and Arabidopsis. Developed hairy roots were compared with intact roots to characterize the gene response to aluminum, NaCl stress and cellular localization of genes. Aluminum-responsive orthologous genes function could be evaluated by using NtSTOP1-KD and Atstop1 as a host for hairy roots transformation.