A rapid and efficient in vitro regeneration system for lettuce (Lactuca sativa L.)

Isabel Armas,Natalia Pogrebnyak,Ilya Raskin

doi:10.1186/s13007-017-0208-0

Isabel Armas, Natalia Pogrebnyak + Show 1 more

Open Access

https://doi.org/10.1186/s13007-017-0208-0

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Journal: Plant methods	Publication Date: Jul 21, 2017
Citations: 20	License type: open-access

Affiliation: Rutgers, The State University of New Jersey

Abstract

BackgroundSuccessful biotechnological improvement of crop plants requires a reliable and efficient in vitro regeneration system. Lettuce (Lactuca sativa L.), one the most important vegetable crops worldwide, is strongly genotype-dependent in terms of regeneration capacity, limiting the potential for biotechnological improvement of cultivars which show recalcitrance under currently available protocols. The effect of different nutrient sources, plant hormone combinations and activated charcoal supplementation on shoot induction efficiency was evaluated on the cultivar ‘RSL NFR’, which had previously shown poor regeneration efficiency.ResultsMultiple shoot organogenesis from cotyledon explants was recorded at the highest frequency and speed on Murashige and Skoog regeneration medium supplemented with 200 mg/l of activated charcoal, 3% sucrose, 10 mg/l benzylaminopurine and 0.5 mg/l naphthaleneacetic acid, which induced shoots through direct regeneration in 90.8 ± 7.9% of explants. High shoot induction efficiency was also observed, albeit not quantified, when using this medium on some other cultivars.ConclusionsThis activated charcoal-containing regeneration medium might offer a rapid and efficient option for direct shoot induction in some lettuce genotypes that do not respond well to common lettuce regeneration protocols. This is also the first report of the effect of activated charcoal in lettuce tissue culture.

Highlights

Successful biotechnological improvement of crop plants requires a reliable and efficient in vitro regeneration system
Petri dishes were kept inside a GC-96 CW walk-in environmentally controlled growth chamber (EGC, Chagrin Falls, OH, USA) at 22 °C, under a photosynthetically active radiation (PAR) light intensity of 10.6 ± 1.7 mol/ m2 day provided by full spectrum 32W Philips F32T8/ DX fluorescent lamps (Philips, Andover, MA, USA) and a 16/8 h photoperiod
Nutritional and hormonal variations of the shoot induction medium Modification of the salt mix and sugar components of the original medium yielded no significant changes when compared to the control medium (Fig. 1, M1–M2)

Summary

Introduction

Successful biotechnological improvement of crop plants requires a reliable and efficient in vitro regeneration system. Lettuce is a good candidate for agronomic trait improvement, including the delivery of nutrients and bioactive compounds beneficial to human health. Following this line of reasoning, lettuce varieties with traits such as herbicide resistance [2], virus resistance [3], environmental stress resistance [4, 5], yield enhancement [6], accumulation of pharmaceutical proteins [7, 8], increased leaf calcium content [9], and increased total polyphenol content [10, 11], have been developed.

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