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Establishment of effective plantlets regeneration protocol via isolated microspore culture in Malaysian indica rice MR219

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Abstract

The current study recognised the issues encountered in regenerating Malaysia MR219 rice plantlet via microspore culture and attempted to develop an efficient protocol in overcoming the restraints. In the present study, a high proportion of uninucleate microspores (49.17%) was isolated from Stage 2-Segment II panicle (59–61 days), which also exhibited the highest callus initiation rate of 8.50%. Maintenance of the panicles under a cool temperature of 4 °C for 7 days before isolating the microspores, resulted in the highest microspore viability of 58.33% and callus initiation rate of 9.33%. The microspore isolation protocol was also optimised in the present study. The filtration sieve engagement with a pore size of 80 µm and further suspension centrifugation at 800 rpm for 5 min produced the highest microspore viability percentage and callus initiation rate. The incorporation of 3.0 mg/L kinetin in conjunction with 0.5 mg/L 2,4-D greatly enhanced the callus initiation rate, with 11.33%. The callus proliferation capacity, with the formation of 481.67 mg callus, was significantly promoted by the addition of 1.0 mg/L kinetin and 0.5 mg/L 2,4-D into the growth medium. Moreover, a higher green plantlet regeneration frequency of 2.83% was induced by the supplementation of 8% sucrose, which produced an average of 3.50 green plantlets.

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Abbreviations

MS:

Murashige and Skoog

2,4-D:

2,4-Dichlorophenoxyacetic acid

BAP:

6-Benzylaminopurine

NAA:

1-Naphthaleneacetic acid

ABA:

Abscisic acid

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Acknowledgements

This study was supported Ministry of Energy, Science, Technology, Environment and Climate Change Research & Development Fund (MESTECC) with grant numbers of TF FP0214B055 (DSTIN) and Malaysian Agricultural Research and Development Institute (MARDI) staff for supporting all research activities

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Authors and Affiliations

  1. Biotechnology & Nanotechnology Research Centre, Malaysian Agricultural Research and Development Institute (MARDI), MARDI Headquarters, 43400, Serdang, Malaysia

    Zuraida Abd Rahman, Zulkifli Ahmad Seman & Ayu Nazreena Othman

  2. Research Innovation Centre Excellence, Malaysian Agricultural Research and Development Institute (MARDI), 13200 Seberang Perai, Kepala Batas, Penang, Malaysia

    Mohamad Bahagia Ab Ghaffar

  3. Centre for Marker Discovery and Validation, Technology Commercialization and Technical Services Centre MARDI Headquarters, 43400, Serdang, Malaysia

    Shahril Ab Razak, Muhammad Fairuz Mohd Yusof & Khairun Hisam Nasir

  4. Department of Plant Protection, Faculty of Agriculture, Universiti Putra Malaysia, UPM Serdang, 43400, Serdang, Malaysia

    Khairulmazmi Ahmad

  5. School of Biological Science, Universiti Sains Malaysia, 11800, Georgetown, Penang, Malaysia

    Yeow Lit Chow & Sreeramanan Subramaniam

  6. Centre for Research in Biotechnology for Agriculture, Universiti Malaya, 50603 Lembah Pantai, Kuala Lumpur, Malaysia

    Teo Chee How

  7. Plant Biotechnology Centre, Agro-Biotechnology Institute, C/O MARDI Headquarters, 43400, Serdang, Selangor, Malaysia

    Norsharina Md Saad

  8. Centre for Chemical Biology, Universiti Sains Malaysia, 11900, Bayan Lepas, Penang, Malaysia

    Sreeramanan Subramaniam

  9. School of Chemical Engineering Technology, Universiti Malaysia Perlis (UNIMAP), 02600, Arau, Perlis, Malaysia

    Sreeramanan Subramaniam

Authors
  1. Zuraida Abd Rahman
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  2. Zulkifli Ahmad Seman
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  3. Ayu Nazreena Othman
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  4. Mohamad Bahagia Ab Ghaffar
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  5. Shahril Ab Razak
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  6. Muhammad Fairuz Mohd Yusof
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  7. Khairun Hisam Nasir
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  8. Khairulmazmi Ahmad
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  9. Yeow Lit Chow
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  10. Teo Chee How
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  11. Norsharina Md Saad
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  12. Sreeramanan Subramaniam
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Contributions

ZAR, ZAS, ANO, KA were responsible for the investigations and experiments; ZAR, KH were writing the original draft and editing of the manuscript; YLC, SS were responsible for review and editing of the manuscript; ZAR, ZAS, EZ, MBAG and TCH were responsible for methodology and conducting formal analysis; SAR, MFMY were responsible for the visualisation of figures and tables; ZAR, SS were responsible for the resources, review and editing of the manuscript.

Corresponding authors

Correspondence to Zuraida Abd Rahman or Sreeramanan Subramaniam.

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Rahman, Z.A., Seman, Z.A., Othman, A.N. et al. Establishment of effective plantlets regeneration protocol via isolated microspore culture in Malaysian indica rice MR219. Plant Biotechnol Rep 16, 343–355 (2022). https://doi.org/10.1007/s11816-022-00742-4

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  • DOI: https://doi.org/10.1007/s11816-022-00742-4

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