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Development of genetically modified sweet cherry rootstock ‘Gisela 6’ with overexpression of PcMPK3-HA gene by Agrobacterium-mediated genetic transformation

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Abstract

Genetic engineering is an efficient technique to improve target traits and provide a potential impetus for germplasm innovation of cherry rootstocks. In this study, we used cherry rootstock ‘Gisela 6’ (Prunus cerasus × Prunus canescens) as the receptor for Agrobacterium-mediated genetic transformation and introduce the recombinant gene PcMPK3-HA into ‘Gisela 6’ rootstock. In addition, we identified phenotypes during the formation of adventitious roots in transgenic lines. The results showed that a combination of 2.0 mg L−1 6-benzylaminopurine (BAP) and 1.0 mg L−1 indole-3-butyric acid (IBA) effectively induced regeneration of ‘Gisela 6’ leaf explants. Selective pressure was a key factor affecting the efficiency of regeneration and transformation. We obtained 19 kanamycin-resistant buds of cherry rootstock and polymerase chain reaction (PCR) revealed five positive buds. Southern blotting confirmed that the PcMPK3-HA gene was integrated into the genome of transgenic lines. Quantitative reverse transcriptase-PCR (qRT-PCR) confirmed significantly higher expression of the PcMPK3 gene in transgenic lines than in wild-type ‘Gisela 6’. Phenotypic analysis showed that the overexpression of PcMPK3-HA enhanced the formation of adventitious roots in transgenic rootstocks, and exerted a positive effect on root growth in tissue-cultured rootstocks following transplantation. Altogether, new germplasms of cherry rootstocks overexpressing PcMPK3-HA were obtained using Agrobacterium-mediated genetic transformation method.

Key message

New germplasms of cherry rootstocks 6 overexpressing the recombinant gene PcMPK3-HA were obtained.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 32172534), Shandong Provincial Natural Science Foundation (CN) (ZR2021MC094, ZR2020MC119, ZR2020MC133) and Modern Agricultural Technology Industry System of China (CARS-30-1-08). The authors also would like to thank TopEdit (www.topeditsci.com) for its linguistic assistance during the preparation of this manuscript.

Funding

Funding was provided by National Natural Science Foundation of China (Grant No. 32172534) [Natural Science Foundation of Shandong Province (Grant No. ZR2021MC094), Natural Science Foundation of Shandong Province (Grant No. ZR2020MC119), Natural Science Foundation of Shandong Province (Grant No. ZR2020MC133) and Modern Agricultural Technology Industry System of China (Grant No. CARS-30-1-08)].

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Authors and Affiliations

  1. Shandong Institute of Pomology, Shandong Academy of Agricultural Sciences, Taian, China

    Xiaojuan Zong, Li Xu, Yue Tan & Hairong Wei

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  1. Xiaojuan Zong
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  2. Li Xu
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  3. Yue Tan
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  4. Hairong Wei
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Contributions

XZ: Designing and conducting the experiments, Writing – original draft, Investigation. LX: Writing – review and editing. YT: Writing – review and editing. HW: Writing – original draft, Investigation.

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Correspondence to Xiaojuan Zong or Hairong Wei.

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Zong, X., Xu, L., Tan, Y. et al. Development of genetically modified sweet cherry rootstock ‘Gisela 6’ with overexpression of PcMPK3-HA gene by Agrobacterium-mediated genetic transformation. Plant Cell Tiss Organ Cult 151, 375–384 (2022). https://doi.org/10.1007/s11240-022-02359-5

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