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Agrobacterium-mediated transformation of purple raspberry (Rubus occidentalis × R. idaeus) with the PtFIT (FER-like iron deficiency–induced transcription factor 1) gene

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Abstract

An Agrobacterium-mediated transformation protocol for the purple raspberry (R. occidentalis × R. idaeus) ‘Amethyst’ was developed. Using the system, the FER-like iron deficiency–induced transcription factor 1 gene cloned from Populus tremula (PtFIT) was expressed in transgenic raspberry plants. Effects of four inoculum densities and two co-cultivation times on ‘Amethyst’ transformation were tested in two separate experiments. Results showed that an average transformation frequency of 3.9% was achieved under the conditions of 25 mg L−1 kanamycin selection, 3-d co-cultivation, and OD600 0.3 to 0.55 inoculum density. A total of 12 PtFIT-transgenic lines of ‘Amethyst’ were verified using polymerase chain reaction (PCR) analysis. Expression of the PtFIT gene in transgenic lines was evaluated under the iron deficiency or sufficiency condition using the real-time quantitative PCR (RT-PCR); however, the expression showed an inconsistent response to iron deficiency among different transgenic lines. An established transformation system could provide a research tool used to understand gene functions and trait development in raspberry; therefore, the present research will be beneficial to breeding and germplasm improvement of raspberry or other Rubus species.

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Authors and Affiliations

  1. Department of Plant Sciences, North Dakota State University, PO Box 6050, Fargo, ND, 767058108-6050, USA

    Changhyeon Kim & Wenhao Dai

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  1. Changhyeon Kim
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  2. Wenhao Dai
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Correspondence to Wenhao Dai.

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Editor: Baochun Li

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Kim, C., Dai, W. Agrobacterium-mediated transformation of purple raspberry (Rubus occidentalis × R. idaeus) with the PtFIT (FER-like iron deficiency–induced transcription factor 1) gene. In Vitro Cell.Dev.Biol.-Plant 58, 343–350 (2022). https://doi.org/10.1007/s11627-021-10228-7

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  • DOI: https://doi.org/10.1007/s11627-021-10228-7

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