An efficient regeneration protocol for in vitro direct organogenesis in einkorn (Triticum monococcum L.) wheat

Günce Şahi̇n,Mehmet Örgeç,Nusret Zenci̇rci̇

doi:10.15832/ankutbd.891812

Abstract

Coleoptile, leaf and root explants of the einkorn (Triticum monococcum ssp. monococcum) were cultured in vitro to obtain an efficient plant regeneration protocol through direct shoot formation by using different combination and concentration of various plant growth regulators. A total of 180 different auxin and cytokinin combinations were tested for regeneration. Shoot formation was not observed with the root and leaf explants. Shoot formation was obtained only from the coleoptile explants, with a mean of 1.20 ± 0.24 shoots/explant and 86.60% of shoot formation frequency and with a 1.20 ± 0.53 shoots/explant and 80.00% shoot formation frequency on medium supplemented with 0.5 mg L-1 TDZ and 1 mg L-1 TDZ plus 1 mg L-1 NAA, respectively. The shoots were subculture on the MS medium containing the most effective hormone combination concurrently continued to shoot and root formation for 45 days. It was noteworthy that 3.66 ± 0.66 shoots per explant were induced by MS, which contained 1 mg L-1 TDZ plus 1 mg L-1 NAA and 2.0 mg L-1 KIN plus 0.5 mg L-1 NAA during a 45 days. Of the different auxin concentrations tested for rooting, 2.0 mg L-1 IAA was predominant, with the greatest number of roots (12.33 ± 0.88) produced per regenerated shoot. Finally, these well-developed plantlets were acclimatized with a 100 % success rate and they transferred to the ex vitro conditions. A highly efficient regeneration protocol for einkorn wheat was developed using somatic tissue as an explant source for the first time.

Highlights

This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record
Using somatic tissues as an explant source makes it possible to obtain a great amount of material regardless of these short-comings, ventures to determine a reliable plant regeneration protocol using somatic cell cultures for diploid wheat species did not produce positive results (Alikina et al 2016)
Örgeç et al (2021) found that coleoptile explants showed the greatest performance for callus induction and indirect plant regeneration compared with root and leaf explants

Summary

Introduction

This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. The standard technique entails the cultivation of immature tissues on 2,4-D-containing media in the dark, followed by plant differentiation in the light on media devoid of phytohormones (Fennell et al 1996; Tamas et al 2004; Chauhan et al 2007; Miroshnichenko et al 2016) This conventional protocol is often ineffective for many wheat genotypes due to the inability to regenerate entire plants on a regular basis. The objective of this study was to investigate a highly effective in vitro regeneration protocol for the einkorn wheat via adventitious shoot formation from the root, coleoptile, and leaf explants cultured on the MS medium containing different combinations and concentrations of plant growth regulators for the first time. Direct shoot formation from somatic tissue is a remarkable feature of this regeneration protocol

Objectives

Methods

Results