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Establishment of Agrobacterium rhizogenes -mediated hairy root transformation of Crocus sativus L.
3 Biotech ( IF 2.6 ) Pub Date : 2021-01-19 , DOI: 10.1007/s13205-020-02626-2
Shilpi Sharma 1 , Yeshveer Singh 2 , Praveen K Verma 2 , Jyoti Vakhlu 1
Affiliation  

Efficient transformation system for genetic improvement is essential in Crocus sativus, as it lacks sexual reproduction. This is the first report wherein an efficient protocol is developed for the transformation of Crocus sativus L. by Agrobacterium rhizogenes 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 C. sativus that can be further used for gene transfer or gene editing in Crocus for its genetic improvement.



中文翻译:

发根农杆菌介导番红花毛状根转化的建立。

藏红花缺乏有性生殖,因此有效的遗传改良转化系统至关重要这是第一篇开发出发根农杆菌ARqua1高效转化藏红花的方案,转化效率为78.51%。使用含有 Ri 质粒和二元载体质粒 pSITE-4NB 以及红色荧光蛋白 (RFP) 标记基因和 β-葡萄糖醛酸酶 (GUS) 报告基因的 ARqua1 菌株进行选择。接种后 21 天后,通过 RFP 信号、GUS 报告基因测定和测试样品的聚合酶链反应 (PCR) 分析证实转化。这些结果证实了番红花毛状根转化方案的建立,该方案可进一步用于番红花的基因转移或基因编辑,以实现其遗传改良。

更新日期:2021-01-19
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