The search for inhibitors of somatic embryogenesis in Medicago truncatula

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Abstract

The success of protocols for the genetic transformation of legumes is limited by their low ability to regenerate. Plant regeneration can occur both along the path of shoot regeneration and their further rooting, and along the path of somatic embryogenesis (SE). SE is similar to zygotic embryogenesis (ZE). It is a method of asexual reproduction, in which the somatic cell, due to its totipotency, switches on the embryogenesis program.

SE and ZE involve common participants in transcriptional, hormonal, and epigenetic control. Like many processes in the plant organism, SE is controlled by the activity of various stimulants or repressors.

As a result of transcriptional analysis of embryogenic and non-embryogenic calli of Medicago truncatula at different stages of development, putative genes-inhibitors of SE were found. Using the Golden Gate system, we created vectors for agrobacterial transformation to overexpress genes of interest in embryogenic calli and evaluate their effect on SE. Overexpression of two genes encoding the transcription factors from WRKY and Homobox-WOX families had a significant inhibitory effect on SE (the average number of somatic embryos per callus decreased).

The MtCLE16, a previously found SE inhibitor from the CLE peptides group, was edited and heterozygous frameshift mutants were obtained. The impact of loss of its function on the SE capacity remains to be analyzed.

This research was supported by the Sirius University of Science and Technology, project PBB-RND-2243.

Full Text

The success of protocols for the genetic transformation of legumes is limited by their low ability to regenerate. Plant regeneration can occur both along the path of shoot regeneration and their further rooting, and along the path of somatic embryogenesis (SE). SE is similar to zygotic embryogenesis (ZE). It is a method of asexual reproduction, in which the somatic cell, due to its totipotency, switches on the embryogenesis program.

SE and ZE involve common participants in transcriptional, hormonal, and epigenetic control. Like many processes in the plant organism, SE is controlled by the activity of various stimulants or repressors.

As a result of transcriptional analysis of embryogenic and non-embryogenic calli of Medicago truncatula at different stages of development, putative genes-inhibitors of SE were found. Using the Golden Gate system, we created vectors for agrobacterial transformation to overexpress genes of interest in embryogenic calli and evaluate their effect on SE. Overexpression of two genes encoding the transcription factors from WRKY and Homobox-WOX families had a significant inhibitory effect on SE (the average number of somatic embryos per callus decreased).

The MtCLE16, a previously found SE inhibitor from the CLE peptides group, was edited and heterozygous frameshift mutants were obtained. The impact of loss of its function on the SE capacity remains to be analyzed.

This research was supported by the Sirius University of Science and Technology, project PBB-RND-2243.

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About the authors

Zakhar S. Konstantinov

Sirius University

Email: zakhar.konstantinov@mail.ru
ORCID iD: 0000-0002-0825-9418

student

Russian Federation, Sochi

Varvara E. Tvorogova

Saint Peterburg State University

Email: v.tvorogova@spbu.ru
ORCID iD: 0000-0002-0549-1457

cand. sci (biol.), senior researcher, department of genetics and biotechnology

Russian Federation, Saint Petersburg

Elina A. Potsenkovskaia

Sirius University

Email: epots556@gmail.com
ORCID iD: 0000-0002-5045-2641

specialist in plant biology and biotechnology

Russian Federation, Sochi

Ludmila A. Lutova

Saint Peterburg State University

Author for correspondence.
Email: l.lutova@spbu.ru
ORCID iD: 0000-0001-6125-0757

dr. sci. (biol.), professor

Russian Federation, Saint Petersburg

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