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Glucosinolates (GSLs) are plant secondary metabolites comprising sulfur and nitrogen mainly found in plants from the order of Brassicales, such as broccoli, cabbage, and Arabidopsis thaliana . The activated forms of GSL play important roles in fighting against pathogens and have health benefits to humans. The increasing amount of data on A. thaliana generated from various omics technologies can be investigated more deeply in search of new genes or compounds involved in GSL biosynthesis and metabolism. This review describes a comprehensive inventory of A. thaliana GSLs identified from published literature and databases such as KNApSAcK, KEGG, and AraCyc. A total of 113 GSL genes encoding for 23 transcription components, 85 enzymes, and five protein transporters were experimentally characterized in the past two decades. Continuous efforts are still on going to identify all molecules related to the production of GSLs. A manually curated database known as SuCCombase (http://plant-scc.org) was developed to serve as a comprehensive GSL inventory. Realizing lack of information on the regulation of GSL biosynthesis and degradation mechanisms, this review also includes relevant information and their connections with crosstalk among various factors, such as light, sulfur metabolism, and nitrogen metabolism, not only in A. thaliana but also in other crucifers.

Glucosinolates are secondary metabolites that play important roles in plant defense and human health, as their production in plants is enhanced by overexpressing transcription factors. Here, four cabbage transcription factors (IQD1-1, IQD1-2, MYB29-1, and MYB29-2) affecting genes in both aliphatic and indolic glucosinolates biosynthetic pathways and increasing glucosinolates accumulation were overexpressed in watercress. Five IQD1-1, six IQD1-2, five MYB29-1, six MYB29-2, and one GUS hairy root lines were created. The expression of all genes involved in glucosinolates biosynthesis was higher in transgenic lines than in the GUS hairy root line, in agreement with total glucosinolates contents, determined by high-performance liquid chromatography. In transgenic IQD1-1 (1), IQD1-2 (4), MYB29-1 (2), and MYB29-2 (1) hairy root lines, total glucosinolates were 3.39-, 3.04-, 2.58-, and 4.69-fold higher than those in the GUS hairy root lines, respectively. These results suggest a central regulatory function for IQD1-1, IQD1-2, MYB29-1, and MYB29-2 transcription factors in glucosinolates biosynthesis in watercress hairy roots.

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.

Selenium (Se)-enriched broccoli has health-beneficial selenium-containing compounds, but it may contain reduced amounts of chemopreventive glucosinolates. To investigate the basis by which Se treatment influences glucosinolate levels, we treated two broccoli cultivars with 25 μM Na 2 SeO 4 . We found that Se supplementation suppressed the accumulation of total glucosinolates, particularly glucoraphanin, the direct precursor of a potent anticancer compound, in broccoli florets and leaves. We showed that the suppression was not associated with plant sulfur nutrition. The levels of the glucosinolate precursors methionine and phenylalanine as well as the expression of genes involved in glucosinolate biosynthesis were greatly decreased following Se supplementation. Comparative proteomic analysis identified proteins in multiple metabolic and cellular processes that were greatly affected and detected an enzyme affecting methionine biosynthesis that was reduced in the Se-biofortified broccoli. These results indicate that Se-conferred glucosinolate reduction is associated with negative effects on precursor amino acid biosynthesis and glucosinolate-biosynthetic-gene expression and provide information for a better understanding of glucosinolate accumulation in response to Se supplementation in broccoli.

Glucosinolates and their breakdown products are well-known for their cancer-chemoprotective functions and biocidal activities against pathogens and generalist herbivores. Insecticides are commonly used in the production of pak choi (Brassica rapa L. ssp. chinensis var. communis [N. Tsen & S.H. Lee] Hanelt). We studied the effects of four commonly used insecticides, namely, β-cypermethrin, acephate, pymetrozine, and imidacloprid, on glucosinolate metabolism in pak choi. All insecticides significantly increased both the transcription of glucosinolate biosynthetic genes and the aliphatic and total glucosinolate accumulations in pak choi. β-Cypermethrin and acephate caused gradual and continuous up-regulation of gene expression from 0.5 to 24 h after treatment, whereas pymetrozine and imidacloprid did so more rapidly, reaching a peak at 1 h and returning to normal at 3 h. Our findings indicate that the four insecticides affect glucosinolate metabolism in pak choi plants to various degrees and suggest that glucosinolates may be involved in plant insecticide metabolism.

Methyl jasmonate spray treatments (250 μM) were utilized to alter glucosinolate composition in the florets of the commercial broccoli F1 hybrids 'Pirate', 'Expo', 'Green Magic', 'Imperial', and 'Gypsy' grown in replicated field plantings in 2009 and 2010. MeJA treatment significantly increased glucoraphanin (11%), gluconasturtiin (59%), and neoglucobrassicin (248%) concentrations and their hydrolysis products including sulforaphane (152%), phenethyl isothiocyanate (318%), N-methoxyindole-3-carbinol (313%), and neoascorbigen (232%) extracted from florets of these genotypes over two seasons. Increased quinone reductase (QR) activity was significantly correlated with increased levels of sulforaphane, N-methoxyindole-3-carbinol, and neoascorbigen. Partitioning experiment-wide trait variances indicated that the variability in concentrations of sulforaphane (29%), neoascorbigen (48%), and QR activity (72%) was influenced by year-associated weather variables, whereas variation in neoglucobrassicin (63%) and N-methoxyindole-3-carbinol (46%) concentrations was primarily attributed to methyl jasmonate treatment. These results suggest that methyl jasmonate treatment can enhance QR inducing activity by increased hydrolysis of glucoraphanin into sulforaphane and the hydrolysis products of neoglucobrassicin.

Broccoli (Brassica oleracea L. var. italica) is a rich source of chemopreventive compounds. Here, we evaluated and compared the effect of selenium (Se) treatment on the accumulation of anticancer compounds Se-methylselenocysteine (SeMSCys) and glucosinolates in broccoli sprouts and florets. Total Se and SeMSCys content in sprouts increased concomitantly with increasing Se doses. Selenate was superior to selenite in inducing total Se accumulation, but selenite is equally effective as selenate in promoting SeMSCys synthesis in sprouts. Increasing sulfur doses reduced total Se and SeMSCys content in sprouts treated with selenate, but not in those with selenite. Examination of five broccoli cultivars reveals that sprouts generally have better fractional ability than florets to convert inorganic Se into SeMSCys. Distinctive glucosinolate profiles between sprouts and florets were observed, and sprouts contained approximately 6-fold more glucoraphanin than florets. In contrast to florets, glucosinolate content was not affected by Se treatment in sprouts. Thus, Se-enriched broccoli sprouts are excellent for simultaneous accumulation of chemopreventive compounds SeMSCys and glucoraphanin.

In this manuscript, a fast and accurate identification and quantitation by mass spectrometry of indolic glucosinolates and camalexin involved in defense in Arabidopsis thaliana are described. Two elicitation systems, inoculation with Botrytis cinerea and treatment with AgNO(3), were used in Col-0 wild-type and mutant genotypes impaired in the biosynthesis of the selected metabolites. Identification of analytes was carried out by nontargeted LC/ESI-QTOF-MS profiling. Confirmation of indolic glucosinolates and camalexin was achieved by their absence in the cyp79B2/B3 and pad3 mutants as well as their respective fragmentation upon collision-induced dissociation. Camalexin accumulation was induced only after AgNO(3) treatment, whereas all indolic glucosinolates were constitutively present. Inoculation with Botrytis did not influence camalexin concentration but caused most aliphatic and indolic glucosinolates contents to decrease. Only the pen 3.1 mutant showed increased indolic glucosinolate levels after Botrytis or AgNO(3) treatments. In addition, profiles of secondary metabolite in nontreated Col-0 and mutant plants were analyzed by means of partial least squares coupled to discriminant analysis (PLS-DA), and differences in the basal levels of indolic glucosinolates and tryptophan between cyp79B2/B3 plants and the rest of genotypes, including Col-0, were found. This probably has to be taken into consideration when comparing stress responses of Col-0 and cyp79B2/B3. The use of mutants carrying alterations in biosynthetic pathways is proposed as a useful strategy to identify secondary metabolites.