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An efficient protocol for in vitro propagation of the wild legume Cicer microphyllum Benth., a crop wild relative of chickpea (Cicer arietinum L.)

  • Plant Tissue Culture
  • Published:
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Abstract

Cicer microphyllum Benth. is a wild legume that is adapted to the extremely adverse climatic conditions of the cold Himalayan deserts. This rare species is a crop wild relative (CWR) that could be used for chickpea (Cicer arietinum L.) breeding programs to improve abiotic stress tolerance. However, the availability of C. microphyllum biological material is very limited due to its high altitude and remote location. These factors make in vitro plant tissue culture an excellent alternative for the conservation and propagation of this species. In this study, an efficient and reproducible in vitro regeneration protocol was established for C. microphyllum. This process enabled the regeneration of multiple shoots through indirect organogenesis from excised embryos. In addition, the effects of combinations of auxin and cytokinin concentrations in Murashige and Skoog (MS) medium were tested. The highest percentage of callus induction was observed on MS with α-Naphthaleneacetic acid (NAA) and 6-Benzylaminopurine (BAP), and the addition of AgNO3 to MS with NAA and BAP significantly improved shoot morphogenesis. The highest root percentage was observed on half-strength MS with Indole-3-butyric acid (IBA). Regenerated plantlets had a 65% survival rate 25 d after transplanting to garden soil, vermiculite, and vermicompost (1:1:1) potting mix. This protocol can be routinely used in large-scale propagation and possibly in germplasm conservation of this rare and important CWR species for additional use in chickpea breeding programs.

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Acknowledgements

The authors are grateful to the DRDO HQ, Ministry of Defense, Government of India, for providing the necessary facilities and financial assistance. The DIHAR (Leh, India) is duly acknowledged for providing logistic support during the collection of C. microphyllum seeds.

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Authors and Affiliations

  1. Centro de Química de Vila Real (CQ-VR), Universidade de Trás-os-Montes e Alto Douro, 5000-801, Vila Real, Portugal

    Rupesh Kumar Singh

  2. Directorate of Onion and Garlic Research, Rajgurunagar, Pune, Maharashtra, 410505, India

    Sivalingam Anandhan

  3. Centro Interdisciplinario de Investigación para el Desarrollo Integral Regional Unidad Sinaloa (CIIDIR-IPN Unidad Sinaloa), Laboratorio de Fitomejoramiento Molecular, Instituto Politécnico Nacional, Blvd. Juan de Dios Bátiz Paredes no. 250, Col. San Joachín, C.P, 81101, Guasave, Sinaloa, Mexico

    Luz María García-Pérez, Eusebio Nava Pérez & Francisco Roberto Quiroz-Figueroa

  4. Red de Estudios Moleculares Avanzados,Cluster BioMimic®, Instituto de Ecología A. C, Carretera Antigua a Coatepec 351, Congregación el Haya, CP, 91070, Xalapa, Veracruz, Mexico

    Eliel Ruiz-May

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  1. Rupesh Kumar Singh
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  2. Sivalingam Anandhan
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Correspondence to Francisco Roberto Quiroz-Figueroa.

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Singh, R.K., Anandhan, S., García-Pérez, L.M. et al. An efficient protocol for in vitro propagation of the wild legume Cicer microphyllum Benth., a crop wild relative of chickpea (Cicer arietinum L.). In Vitro Cell.Dev.Biol.-Plant 55, 9–14 (2019). https://doi.org/10.1007/s11627-018-09958-y

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  • DOI: https://doi.org/10.1007/s11627-018-09958-y

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