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Efficient micropropagation protocol and genome size estimation of an important cover crop, Mucuna bracteata DC. ex Kurz

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Abstract

Mucuna bracteata DC. ex Kurz is an important cover crop in plantations across the tropics. However, low germination rate and poor viability of Mucuna seeds pose significant challenges of using the seeds as starting material. To address these limitations, we have optimized seed germination conditions (such as scarification period, surface sterilization protocols and imbibition period) and in vitro propagation protocols for M. bracteata. We found that seeds treated with sulphuric acid for 30 min, imbibed for 6 h and incubated in dark conditions on a wet cotton roll (10 mL of sterile distilled water) supplemented with 0.1% activated charcoal produced the highest percentage of seed germination (44%) and seed vigor index. In vitro-derived cotyledonary nodes showed the highest number of shoots per explant (5.60) and rooting response (92.9%) when cultured on Murashige and Skoog medium containing 4.44 µM 6-benzylaminopurine and 10.7 µM 1-naphthaleneacetic acid, respectively. Of the 100 rooted plantlets acclimatized, 89.0% survived after 4 weeks of transplanting. Single sequence repeat and flow cytometry analysis were performed to confirm the genetic fidelity of the plants. Our protocol offers, for the first time, a simple and effective seed germination and scalable propagation procedures for M. bracteata. Furthermore, we have also estimated the genome size (1448 ± 9 Mb) and DNA content (1.48 ± 0.01 pg) for M. bracteata that can be used for future cytogenetic studies on genetic diversity and gene exchange.

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Acknowledgements

The authors would like to acknowledge Mr. Mohd Razik Midin and Dr. Maria Madon from the Advanced Biotechnology and Breeding Centre of Malaysian Palm Oil Board (MPOB), Malaysia, for their technical assistance; the University of Malaya Bright Sparks Programme (BSP/APP/2022/2014), University of Malaya PPP Grant (Grant No: PG254-2016A), High Impact Research Programme of University of Malaya, Malaysia (UM.C/625/1/HIR/MOHE/SCI/18) and CEBAR RU (RU006-2017).

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Authors and Affiliations

  1. Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia

    Nazrin Abd Aziz, Rofina Yasmin Othman & Norzulaani Khalid

  2. Centre for Research in Biotechnology for Agriculture, University of Malaya, 50603, Kuala Lumpur, Malaysia

    Boon Chin Tan, Rofina Yasmin Othman & Norzulaani Khalid

  3. High Impact Research (HIR), University of Malaya, 50603, Kuala Lumpur, Malaysia

    Rofina Yasmin Othman

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  1. Nazrin Abd Aziz
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  2. Boon Chin Tan
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  4. Norzulaani Khalid
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NAA conducted the experiments and analyzed data; NAA, BCT, RYO, NK conceived the idea, designed the experiments and wrote the paper. All authors read and approved the final manuscript.

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Correspondence to Norzulaani Khalid.

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Communicated by Sergio J. Ochatt.

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Abd Aziz, N., Tan, B.C., Othman, R.Y. et al. Efficient micropropagation protocol and genome size estimation of an important cover crop, Mucuna bracteata DC. ex Kurz. Plant Cell Tiss Organ Cult 132, 267–278 (2018). https://doi.org/10.1007/s11240-017-1376-3

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