Main conclusion Glucosinolates are biologically active compounds which are involved in plant defense reaction. The use of plant in vitro cultures and genetic engineering is a promising strategy for their sustainable production.Glucosinolates are a class of secondary metabolites found mainly in Brassicaceae, which contain nitrogen and sulfur in their structures. Glucosinolates are divided into three groups depending on the amino acid from which they are biosynthesized. Aliphatic glucosinolates are generally derived from leucine, valine, methionine, isoleucine and alanine while indole and aromatic glucosinolates are derived from tryptophan and phenylalanine or tyrosine, respectively. These compounds are hydrolyzed by the enzyme myrosinase when plants are stressed by biotic and abiotic factors, obtaining different degradation products. Glucosinolates and their hydrolysis products play an important role in plant defense responses against different types of stresses. In addition, these compounds have beneficial effect on human health because they are strong antioxidants and they have potent cardiovascular, antidiabetic, antimicrobial and antitumoral activities. Due to all the properties described above, the demand for glucosinolates and their hydrolysis products has enormously increased, and therefore, new strategies that allow the production of these compounds to be improved are needed. The use of plant in vitro cultures is emerging as a biotechnological strategy to obtain glucosinolates and their derivatives. This work is focused on the biosynthesis of glucosinolates and the bioactivity of these compounds in plants. In addition, a detailed study on the strategies used to increase the production of several glucosinolates, in particular those synthesized in Brassicaceae, using in vitro plant cultures has been made. Special attention has been paid for increasing the production of glucosinolates and their derivatives using metabolic engineering.

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In this work, the effect of different elicitors and culture conditions on the production of glucosinolates in broccoli cell cultures was studied. The results showed that 0.5 μM coronatine was the best elicitor for increasing glucosinolate production (205-fold increase over untreated cells after 72 h of treatment). Furthermore, the expression levels of some genes related to the biosynthetic pathway of glucosinolates as well as three Myb transcription factors also have been studied. The highest glucosinolate levels found in coronatine-treated cells were closely correlated with the highest gene expression levels of Cyp79b2, Cyp83b1, St5a, Myb51, and Myb122 after 6 h of treatment. The data shown in this study provide new insight into the key metabolic steps involved in the biosynthesis of glucosinolates, which will be of use for future applications of metabolic engineering techniques in broccoli.

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