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建立农杆菌介导的L.毛状根转化体系

Establishment of -mediated hairy root transformation of L.

作者信息

Sharma Shilpi, Singh Yeshveer, Verma Praveen K, Vakhlu Jyoti

机构信息

School of Biotechnology, University of Jammu, Jammu, Jammu and Kashmir 180006 India.

National Institute of Plant Genome Research (NIPGR), Aruna Asaf Ali Marg, New Delhi, 110067 India.

出版信息

3 Biotech. 2021 Feb;11(2):82. doi: 10.1007/s13205-020-02626-2. Epub 2021 Jan 19.

DOI:10.1007/s13205-020-02626-2
PMID:33505837
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7813919/
Abstract

Efficient transformation system for genetic improvement is essential in as it lacks sexual reproduction This is the first report wherein an efficient protocol is developed for the transformation of L. by strain ARqua1 with a transformation efficiency of 78.51%. The ARqua1 strain harboring both Ri plasmid and binary vector plasmid pSITE-4NB, and marker genes for red fluorescent protein (RFP) and a β-glucuronidase (GUS) reporter gene were used for selection. Transformation was confirmed by RFP signal, GUS reporter assay and polymerase chain reaction (PCR) analysis of the test samples after 21 days post inoculation. These results confirm the establishment of protocol for hairy root transformation in that can be further used for gene transfer or gene editing in for its genetic improvement.

摘要

由于缺乏有性繁殖,高效的遗传改良转化系统对于[具体植物名称未给出]至关重要。这是第一份关于开发高效转化方案的报告,该方案用于通过ARqua1菌株转化[具体植物名称未给出],转化效率为78.51%。携带Ri质粒和二元载体质粒pSITE - 4NB的ARqua1菌株,以及红色荧光蛋白(RFP)和β - 葡萄糖醛酸酶(GUS)报告基因的标记基因用于筛选。接种后21天,通过RFP信号、GUS报告基因检测和测试样品的聚合酶链反应(PCR)分析确认转化。这些结果证实了[具体植物名称未给出]毛状根转化方案的建立,该方案可进一步用于[具体植物名称未给出]的基因转移或基因编辑以实现其遗传改良。

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本文引用的文献

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Development of a system for efficient callus production, somatic embryogenesis and gene editing using CRISPR/Cas9 in Saffron ( L.).利用CRISPR/Cas9技术开发一种用于藏红花高效愈伤组织生产、体细胞胚胎发生和基因编辑的系统。
Plant Methods. 2020 Apr 7;16:47. doi: 10.1186/s13007-020-00589-2. eCollection 2020.
2
An efficient and reproducible -mediated transformation method for hexaploid wheat ( L.).一种用于六倍体小麦(L.)的高效且可重复的介导转化方法。
Plant Methods. 2019 Oct 26;15:121. doi: 10.1186/s13007-019-0503-z. eCollection 2019.
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A critical review on use of Agrobacterium rhizogenes and their associated binary vectors for plant transformation.根癌农杆菌及其相关二元载体在植物转化中的应用的批判性评价。
Biotechnol Adv. 2019 Nov 15;37(7):107405. doi: 10.1016/j.biotechadv.2019.06.004. Epub 2019 Jun 8.
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-mediated hairy roots transformation as a tool for exploring aluminum-responsive genes function.介导的毛状根转化作为探索铝响应基因功能的工具。
Future Sci OA. 2019 Feb 8;5(3):FSO364. doi: 10.4155/fsoa-2018-0065. eCollection 2019 Mar.
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Optimization of Agrobacterium-mediated transformation in spring bread wheat using mature and immature embryos.利用成熟胚和未成熟胚优化农杆菌介导的春小麦转化
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