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Optimization of an indirect regeneration system for common bean (Phaseolus vulgaris L.)

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Abstract

The main problems associated with growing common bean (Phaseolus vulgaris L.) callus are low proliferation and differentiation, and high browning. In this study, common bean callus induced by cotyledon nodes was used as explant material to investigate the effects of different exogenous substances on callus regeneration, as well as the correlation between callus browning and the changes in superoxide dismutase (SOD), peroxidase (POD), and polyphenol oxidase (PPO) enzyme activities during callus culture. Adding AgNO3, CoCl2, Putrescine (Put), and 1-aminocyclopropane-1-carboxylic acid (ACC) to the callus growth medium at appropriate concentrations could significantly improve callus proliferation and differentiation efficiency, while also reducing the degree of browning, compared to controls. Callus explants were cultured in callus proliferation and differentiation medium (CPADM) containing 5 mg·L−1 AgNO3, 15 mg·L−1 Put, 5 mg·L−1 CoCl2, or 0.02 mM ACC, with optimal callus growth at these dosages. After treatment with the four exogenous substances, callus browning was inversely correlated with SOD activity but positively correlated with POD and PPO activities. The maximum rooting frequency of shoots was observed when rooting media was treated with 1 mg·L−1 Indole butyric acid (IBA) or 0.1 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D). The average number of primary roots, root length, and root fresh weight were higher after these treatments. The indirect regeneration issue was largely resolved for common bean by the callus culture technique applied in this study, which provides a framework for genetic modification, germplasm preservation, and bean application.

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Data availability

The data supporting the findings of this study are available from the authors upon reasonable request.

Abbreviations

AgNO3 :

Silver nitrate

Put:

Putrescine

CoCl2 :

Cobalt chloride

ACC:

1-Aminocyclopropane-1-carboxylic acid

TDZ:

Thioridazine

IBA:

Indole butyric acid

2,4-D:

2,4-Dichlorophenoxyacetic acid

MS:

Murashige and Skoog

CPADM:

Callus proliferation and differentiation medium (MS, 0.5 mg·L−1 TDZ, 3% sucrose, 0.65% agar)

SOD:

Superoxide dismutase

POD:

Peroxidase

PPO:

Polyphenol oxidase

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Acknowledgements

This study was funded by Heilongjiang Provincial Natural Science Foundation of China (Grant number LH2020C090)

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Author notes

  1. Luxi Xiong and Chang Liu have contributed equally to this work and share the first authorship.

Authors and Affiliations

  1. Horticulture Department, College of Advanced Agriculture and Ecological Environment, Heilongjiang University, Harbin, 150000, China

    Luxi Xiong, Chang Liu, Dajun Liu, Zhishan Yan, Xiaoxu Yang & Guojun Feng

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  1. Luxi Xiong
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  2. Chang Liu
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  3. Dajun Liu
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  4. Zhishan Yan
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Contributions

LXX and CL: performed the experiments, analyzed the results and wrote the draft manuscript. DJL: supervised and directed this work and edited the manuscript. ZSY: provided common bean seeds. XXY: and GJF: conceived the project, supervised the research, and reviewed the manuscript. All authors read and approved the manuscript.

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Correspondence to Xiaoxu Yang or Guojun Feng.

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Xiong, L., Liu, C., Liu, D. et al. Optimization of an indirect regeneration system for common bean (Phaseolus vulgaris L.). Plant Biotechnol Rep 17, 821–833 (2023). https://doi.org/10.1007/s11816-023-00830-z

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