African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12487

Full Length Research Paper

Optimization of regeneration protocol and prospecting spectinomycin resistance in barley (Hordeum vulgare L.) cv Haider-93

Manzar Abbas
  • Manzar Abbas
  • School of Agriculture, Forestry and Food Engineering, Yibin University, Yibin 644000, Sichuan, China.
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Syed Sarfaraz Hussain
  • Syed Sarfaraz Hussain
  • Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China.
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Saqi Kosar Abbas
  • Saqi Kosar Abbas
  • Department of Plant Protection, Ministry of National Food Security and Research, Islamabad, Pakistan.
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Sammar Abbas
  • Sammar Abbas
  • College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing, China.
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Muhammad Tahir
  • Muhammad Tahir
  • Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing, China.
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Yongying Su
  • Yongying Su
  • School of Agriculture, Forestry and Food Engineering, Yibin University, Yibin 644000, Sichuan, China.
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Jia Li
  • Jia Li
  • School of Agriculture, Forestry and Food Engineering, Yibin University, Yibin 644000, Sichuan, China.
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Nisar Ahmed
  • Nisar Ahmed
  • Centre of Agriculture Biochemistry and Biotechnology (CABB), Faculty of Agriculture, University of Agriculture, Faisalabad 38000, Pakistan.
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Muhammad Sarwar Khan
  • Muhammad Sarwar Khan
  • Centre of Agriculture Biochemistry and Biotechnology (CABB), Faculty of Agriculture, University of Agriculture, Faisalabad 38000, Pakistan.
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Caiming Gou
  • Caiming Gou
  • School of Agriculture, Forestry and Food Engineering, Yibin University, Yibin 644000, Sichuan, China.
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  •  Received: 23 June 2023
  •  Accepted: 14 August 2023
  •  Published: 30 September 2023

Abstract

Immature zygotic embryos from spring barley cv. Haider-93 were used to induce somatic embryogenesis. The type of the explant, the level of 2,4-dichlorophenoxyacetic acid (2,4-D) and handling of calli during subculture are critical factors to obtain maximum number of regenerants. Different concentrations of 2,4-D (0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 mg/L) and Kinetin (0.5, 1.0, 1.5, 2.0 mg/L) were used for callus induction and shoot initiation, respectively. Use of immature embryos having damaged axis as explants revealed a pronounced gradient of callus formation on Murashige and Skoog (MS) medium supplemented with 2.5 mg/L of 2,4-D and maximum regeneration response at 1 mg/L kinetin. Regenerated shoots were rooted on MS medium supplemented with 1 mg/L IAA. A kill curve was developed to find out the sensitivity level of barley cells to spectinomycin, a broad-spectrum antibiotic. This study fulfils an antique prerequisite of a reproducible regeneration system required for the improvement of barley via genetic engineering and also falls under, “Establish Good Health and Well-Being” Sustainable Development Goals of United Nations Organization.

 

Key words: 2,4-Dichlorophenoxyacetic acid, indole acetic acid, kinetin, antibiotic resistance, spectinomycin, Barley 53.