Disturbance of ethylene biosynthesis and perception during somatic embryogenesis in <Emphasis Type="Italic">Medicago sativa</Emphasis> L. reduces embryos’ ability to regenerate |
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Authors: | Ewa Kępczyńska Sylwia Zielińska |
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Institution: | (1) Department of Plant Biotechnology, University of Szczecin, Wąska 13, 71-415 Szczecin, Poland |
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Abstract: | Effects of non-specific ethylene biosynthesis inhibitors: salicylic acid (SA) and aminoethoxyvinylglycine (AVG), and of specific
inhibitors of ethylene binding to receptors: 1-methylcyclopropene (1-MCP) and 2,5-norbornadiene (NBD) applied during proliferation
and differentiation phases of indirect somatic embryogenesis (SE) of Medicago sativa L. cv. Rangelander on embryogenic suspension growth, embryo production, development, and ability to germinate and convert
were studied. Application of SA and AVG alone or together at concentrations from 1 to 500 μM in B5g liquid medium during the proliferation phase had an inhibitory effect on ethylene production and embryogenic suspension
growth. Additionally, it caused a drastic reduction in production of embryos and their development on BOi2Y solid differentiation
medium. The inhibitory effect of SA was more visible than that of AVG. In addition, disturbance of ethylene biosynthesis during
the proliferation phase of SE resulted in diminished lateral germination and conversion of cotyledonary embryos on MS solid
medium. Moreover, blocking of ethylene receptors by 1-MCP during the proliferation phase also inhibited ethylene production
and embryogenic suspension growth and reduced embryo production during differentiation. MCP almost completely inhibited development
of cotyledonary embryos. At the same time, development of more embryos was arrested at the globular stage, and the number
of abnormal embryos almost doubled. Similarly, addition of 1-MCP or NBD to the ambient atmosphere during the differentiation
phase evidently arrested the development of embryos and, consequently, their ability to germinate and convert on MS regeneration
medium. All the results presented above demonstrated that not only ethylene biosynthesis, but also ethylene action is involved
in the control of individual phases of SE in Medicago sativa L. cv. Rangelander. And what is more, disturbance of these processes during distinct phases of SE adversely affects vigor
of the somatic embryos obtained. |
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