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Development of an efficient and reproducible regeneration system in wheat (Triticum aestivum L.)

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Abstract

The availability of reproducible regeneration system through tissue culture is a major bottleneck in wheat improvement program. The present study has considered to develop an efficient callus induction and regeneration system using mature and immature embryos as explants in recently released agronomically superior spring wheat varieties. An efficient sterilization process was standardized using 0.1% HgCl2 and 70% ethanol for both seeds and embryos. The maximum possible combinations of plant growth regulators (PGRs) were evaluated for their effect on different wheat regeneration processes through tissue culture starting from callus to root induction. Picloram is found as an effective auxin with 87.63–98.67% callus induction efficiency in both explants. Supplementation of CuSO4 along with 2,4-D, zeatin in regeneration medium significantly enhanced the multiple shoot induction. The shoot development was achieved using full strength Murashige and Skoog’s (MS) medium and root induction using half MS medium without PGRs. The optimized medium and method has resulted up to 100% regeneration irrespective of the genotype used with high reproducibility. Thus, the standardized regeneration system can be used in the regeneration of healthy plants from embryos rescued from interspecies crosses, transgenic production, induced mutation breeding and recently developed genome editing techniques for the procreation of wheat plants having novel traits.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

BAP:

6-Benzylaminopurine

Dicamba:

3,6-Dichloro-2-methoxybenzoic acid

IAA:

Indole-3-acetic acid

NAA:

1-Naphthaleneacetic acid

PGRs:

Plant growth regulators

Picloram:

4-Amino-3,5,6-trichloro-2-pyridinecarboxylic acid

RH:

Relative humidity

TDZ:

Thidiazuron

Zeatin:

6-(4-Hydroxy-3-methylbut-2-enylamino)purine

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Acknowledgements

This work is financially supported by the Indian Council of Agricultural Research, New Delhi, India under the project entitled ICAR Network Project “Transgenic in crops (NPTC)” (Project No. 1006474).

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Authors and Affiliations

  1. Biotechnology Engineering, University Institute of Engineering and Technology, Kurukshetra University, Kurukshetra, Haryana, 136119, India

    Rakesh Kumar, Amandeep Kaur & Anita Grewal

  2. ICAR-Indian Institute of Wheat and Barley Research (IIWBR), Karnal, Haryana, 132001, India

    Rakesh Kumar, Harohalli Masthigowda Mamrutha, Amandeep Kaur, Karnam Venkatesh, Raj Kumar & Vinod Tiwari

Authors
  1. Rakesh Kumar
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  2. Harohalli Masthigowda Mamrutha
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  3. Amandeep Kaur
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  4. Karnam Venkatesh
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  5. Anita Grewal
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  6. Raj Kumar
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  7. Vinod Tiwari
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Contributions

MHM conceived the project and designed the experiments with AG. RK conducted experiments. AK assisted RK during Research. RK, MHM, and KV wrote the manuscript. RajK and VT guided and extended facilities for research. All authors read and approved the final manuscript.

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Correspondence to Harohalli Masthigowda Mamrutha.

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The authors declare that they have no conflict of interest.

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Kumar, R., Mamrutha, H.M., Kaur, A. et al. Development of an efficient and reproducible regeneration system in wheat (Triticum aestivum L.). Physiol Mol Biol Plants 23, 945–954 (2017). https://doi.org/10.1007/s12298-017-0463-6

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  • DOI: https://doi.org/10.1007/s12298-017-0463-6

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