A highly efficient organogenesis protocol based on zeatin riboside for in vitro regeneration of eggplant

Edgar García-Fortea,Vicente Moreno,Benito José Pineda-Chaza,Juan Pablo Bracho-Gil,Ana García-Pérez,Santiago Vilanova,Agustín Lluch-Ruiz,Jaime Prohens,Pietro Gramazio,Mariola Plazas

doi:10.1186/s12870-019-2215-y

Abstract

BackgroundEfficient organogenesis induction in eggplant (Solanum melongena L.) is required for multiple in vitro culture applications. In this work, we aimed at developing a universal protocol for efficient in vitro regeneration of eggplant mainly based on the use of zeatin riboside (ZR). We evaluated the effect of seven combinations of ZR with indoleacetic acid (IAA) for organogenic regeneration in five genetically diverse S. melongena and one S. insanum L. accessions using two photoperiod conditions. In addition, the effect of six different concentrations of indolebutyric acid (IBA) in order to promote rooting was assessed to facilitate subsequent acclimatization of plants. The ploidy level of regenerated plants was studied.ResultsIn a first experiment with accessions MEL1 and MEL3, significant (p < 0.05) differences were observed for the four factors evaluated for organogenesis from cotyledon, hypocotyl and leaf explants, with the best results obtained (9 and 11 shoots for MEL1 and MEL3, respectively) using cotyledon tissue, 16 h light / 8 h dark photoperiod conditions, and medium E6 (2 mg/L of ZR and 0 mg/L of IAA). The best combination of conditions was tested in the other four accessions and confirmed its high regeneration efficiency per explant when using both cotyledon and hypocotyl tissues. The best rooting media was R2 (1 mg/L IBA). The analysis of ploidy level revealed that between 25 and 50% of the regenerated plantlets were tetraploid.ConclusionsAn efficient protocol for organogenesis of both cultivated and wild accessions of eggplant, based on the use of ZR, is proposed. The universal protocol developed may be useful for fostering in vitro culture applications in eggplant requiring regeneration of plants and, in addition, allows developing tetraploid plants without the need of antimitotic chemicals.

Highlights

Efficient organogenesis induction in eggplant (Solanum melongena L.) is required for multiple in vitro culture applications
We developed a new universal regeneration protocol based on the use of zeatin riboside (ZR) that has proven to be highly efficient in eggplant, which is a genotype dependent recalcitrant species to in vitro culture [9,10,11]
We developed a universal protocol for the eggplant regeneration

Summary

Introduction

Efficient organogenesis induction in eggplant (Solanum melongena L.) is required for multiple in vitro culture applications. Common eggplant (Solanum melongena L.), known as brinjal eggplant, is one of the most important vegetables, globally ranking fifth in total production among vegetable crops [1]. This crop has a great interest due to its high content in bioactive compounds, mostly phenolics, that have multiple properties beneficial for human health [2]. The development of new genome editing technologies in plant breeding has generated a growing interest in (2020) 20:6 in vitro culture for regeneration protocols [13, 14]. Tetraploids can be the starting point for obtaining triploids which, in addition to the lack of seeds, can exhibit superior quality parameters in the fruit

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