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Somatic embryogenesis and β-glucuronidase transformation in chickpea (Cicer arietinum cv. Bivanich)

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Abstract

Background

Chickpea (Cicer arietinum L.) is an important kernel legume in the world. To optimize the plant tissue culture some experiments such as direct regeneration, proliferation, rooting shoots and somatic embryogenesis were done.

Methods and results

In experiments were used direct regeneration and proliferation, various levels of plant growth regulators NAA (0 and 1.0 mg/l), BAP (0, 1, 3 and 5 mg/l) and three explants’ types (epicotyl, cotyledon and embryonic axis). The results of both experiments showed that embryonic axis explant was better than other explants. The highest percentage was obtained in MS media containing 1 mg/l BAP and also 3 mg/l BAP and 0.1 mg/l NAA with an average of 72%. The highest average number of branches (4.66) was found in the proliferation of embryonic axis in MS medium containing 3 mg/l BAP. The highest rooting shoot (90%) was found in 1/2MS in B5 medium vitamins with 0.2 mg/l of IBA and 0.5 mg/l NAA. Somatic embryogenesis experiments were compared on the concentration gradient of 2,4-D in fine embryonic axis explants. The results displayed that the concentration gradient of 10 mg/l 2,4-D to 5 mg/l of 2,4-D and then to zero concentration showed the highest number of embryos.

Conclusion

The best environment for regeneration embryos was MS medium with 2.5 mg/l of 2,4-D concentration gradient to zero. In this study, the PCR reaction showed the presence of the β-glucuronidase (gus) marker gene in regenerated cotyledons for 20 min in all three strains studied.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Abbreviations

BAP:

Benzylaminopurine

BA:

Benzyl adenine

NAA:

Naphthaleneacetic acid

MS medium:

Murashige and Skoog medium

PGRs:

Plant growth retardants

IBA:

Indole-3-butyric acid

2,4-D:

2,4-Dichlorophenoxyacetic acid

LB medium:

Lysogeny Broth medium

DDW:

Deuterium-depleted water

PCR:

Polymerase chain reaction

ppm:

Part per million

CTAB:

Cetyltrimethylammonium bromide

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Acknowledgements

The authors would like to thank all those who work in the Department of Agronomy and Plant Breeding, Campus of Agriculture and Natural Resources for their help. Financial support for this work has been provided by Razi University Grants No. 56747.

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No external funding was obtained for this project.

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Authors and Affiliations

  1. Department of Agronomy and Plant Breeding, Razi University, Kermanshah, Iran

    Mohammad Reza Amani, Alireza Zebarjadi & Danial Kahrizi

  2. Faculty of Agriculture, Department of Horticulture, Ataturk University, 25240, Erzurum, Turkey

    Sezai Ercisli

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  1. Mohammad Reza Amani
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Amani, M.R., Zebarjadi, A., Kahrizi, D. et al. Somatic embryogenesis and β-glucuronidase transformation in chickpea (Cicer arietinum cv. Bivanich). Mol Biol Rep 49, 11219–11227 (2022). https://doi.org/10.1007/s11033-022-07450-w

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