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An improved and simplified somatic embryogenesis protocol in Chir pine (Pinus roxburghii).

Farah Kousar1, Ulrika Egertsdotter2, Faheem Aftab1

  • 1Institute of Botany, University of the Punjab, Lahore, 54590, Pakistan.

Scientific Reports
|October 29, 2025
PubMed
Summary

This study details an improved method for somatic embryogenesis in Pinus roxburghii, a vital tree species. The research successfully outlines developmental stages and optimizes media for better embryo production and maturation, crucial for reforestation efforts.

Keywords:
ConiferEmbryogenic tissueMaturationSomatic embryosWestern himalayan mountains

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Area of Science:

  • Forestry
  • Plant Biotechnology
  • Conservation Biology

Background:

  • Forests are crucial for bioeconomy but face global depletion.
  • Pinus roxburghii holds significant ecological and economic value for Pakistan.
  • This species is vital for climate change adaptation through afforestation and reforestation.

Purpose of the Study:

  • To develop an improved protocol for somatic embryogenesis in Pinus roxburghii.
  • To document the developmental stages of zygotic and somatic embryos for this species.
  • To optimize media formulations for enhanced initiation, proliferation, maturation, and conversion of somatic embryos.

Main Methods:

  • Initiation of somatic embryogenesis using immature pre-cotyledonary zygotic embryos on specific media (LP-889).
  • Proliferation and maintenance of embryogenic tissue using optimized media (LP-1250) with plant growth regulators.
  • Maturation of somatic embryos tested on two media (mLV and LP-1562) with varying L-glutamine and abscisic acid (ABA) concentrations.

Main Results:

  • Somatic embryogenesis initiation was influenced by zygotic embryo developmental stage.
  • LP-1562 medium yielded a higher number of somatic embryos (310/g) compared to mLV medium (204/g).
  • Successful conversion of somatic embryos was achieved, but low rooting rates and precocious germination remain challenges.

Conclusions:

  • The study presents a significantly improved protocol for Pinus roxburghii somatic embryogenesis.
  • The findings provide a basis for mass propagation of this ecologically and economically important pine species.
  • Further research is needed to address rooting and precocious germination issues for successful ex-situ conservation and reforestation.