Glucosinolate Production Research Articles

Salinity Tolerance of Diplotaxis tenuifolia Varieties Growing in Spring–Summer Season Under Mediterranean Greenhouse and Optimal Growing Conditions

Salinity is one of the principal abiotic stresses that occurs in the Mediterranean area, causing loss of productivity and decrease of vegetable crop quality. The effect of salinity (0, 25, 75, 150 mM NaCl) was evaluated in three Diplotaxis tenuifolia varieties (Dragon Tongue, Capriccio, Piccante), previously selected for salinity tolerance and high glucosinolates production in leaves. The aim of this research was to explore the salinity tolerance of three wild rocket varieties cultivated under optimal temperature conditions and under high temperature that typically characterized the Mediterranean greenhouse. Biometric, biomass, pigment production and physiological parameters were evaluated. Biometric, physiological, and biochemical parameters significantly varied because of variety, salt level used and environmental conditions. PCA analysis highlighted that the two cultivation systems deeply affected the wild rockets response to salt stress. In general, under optimal growing conditions, wild rocket varieties showed higher growth parameters compared to greenhouse conditions. Overall Capriccio was the most susceptible variety to salinity, while Dragon Tongue (V1) and Piccante (V3) were more tolerant to salt stress. Furthermore, in both growing conditions V1 was the less productive variety while V3 showed an opposite trend. Interestingly, gene (DtOxo and DtGst) expression analysis revealed a significant increase of the target gene expression as response of salinity levels, with a clear increase of DtOxo level in V1 and V3. The results obtained in this study can be useful to plan future breeding programs aimed to increase rocket quality grown under Mediterranean conditions.

Glucosinolates and Their Hydrolytic Products-A Love Story of Environmental, Biological and Chemical Conditions.

Glucosinolates play important role in providing defense to plants and helps them in coping against various biotic as well as abiotic stress. Many living beings including humans, animals, and even some herbivores have adapted themselves to use this defense mechanism for their own use. More than 120 glucosinolates are distributed within a large number of plants. Many factors are known to influence the GSL composition in a plant. Among these, cofactors, myrosinase isozymes, heavy metals and the environmental conditions such as light, CO2 and temperature are important in the regulation. These factors ensure that different glucosinolate composition could be produced by the plants, thus impacting the defense mechanism. The objective of the current review is to highlight the importance of factors responsible for affecting glucosinolate composition and concentration. The review has been compiled using accessible literature from Pubmed, Scopus, and Google scholar. Efforts have been made to restrict the literature to the last 5 years (2018-2023), with some exceptions. The current critical review acts as a resource for all the researchers working on this essential compound. It provides information on the factors that may influence the glucosinolate production. It also gives them an opportunity to modify the glucosinolate composition of a plant using the given information. Glucosinolates have long been an ignored class of biomolecule. The plethora of biological activities of the compounds can be useful. Though there are some harmful components such as goitrin and progoitrin, these can be easily removed by modulating some of the factors highlighted in the review. The current review has touched base on almost all the factors that have the ability to modify glucosinolate composition and concentration. The mechanistic action of these factors have also been discussed using the current available literature.

Fine-Tuning of Arabidopsis thaliana Response to Endophytic Colonization by Gluconacetobacter diazotrophicus PAL5 Revealed by Transcriptomic Analysis.

Gluconacetobacter diazotrophicus is a diazotrophic endophytic bacterium that promotes the growth and development of several plant species. However, the molecular mechanisms activated during plant response to this bacterium remain unclear. Here, we used the RNA-seq approach to understand better the effect of G. diazotrophicus PAL5 on the transcriptome of shoot and root tissues of Arabidopsis thaliana. G. diazotrophicus colonized A. thaliana roots and promoted growth, increasing leaf area and biomass. The transcriptomic analysis revealed several differentially expressed genes (DEGs) between inoculated and non-inoculated plants in the shoot and root tissues. A higher number of DEGs were up-regulated in roots compared to shoots. Genes up-regulated in both shoot and root tissues were associated with nitrogen metabolism, production of glucosinolates and flavonoids, receptor kinases, and transcription factors. In contrast, the main groups of down-regulated genes were associated with pathogenesis-related proteins and heat-shock proteins in both shoot and root tissues. Genes encoding enzymes involved in cell wall biogenesis and modification were down-regulated in shoots and up-regulated in roots. In contrast, genes associated with ROS detoxification were up-regulated in shoots and down-regulated in roots. These results highlight the fine-tuning of the transcriptional regulation of A. thaliana in response to colonization by G. diazotrophicus PAL5.

Myrosinase isogenes in wasabi (Wasabia japonica Matsum) and their putative roles in glucosinolate metabolism

BackgroundWasabi, a Brassicaceae member, is well-known for its unique pungent and hot flavor which is produced from glucosinolate (GSL) degradation. Myrosinase (MYR) is a principle enzyme catalyzing the primary conversion of GSLs to GSL hydrolysis products (GHPs) which is responsible for plant defense system and food quality. Due to the limited information in relation to MYRs present in wasabi (Wasabia japonica M.), this study aimed to identify the MYR isogenes in W. japonica and analyze their roles in relation to GSL metabolism.ResultsIn results, WjMYRI-1 was abundantly expressed in all organs, whereas WjMYRI-2 showed only trace expression levels. WjMYRII was highly expressed in the aboveground tissues. Interestingly, WjMYRII expression was significantly upregulated by certain abiotic factors, such as methyl jasmonate (more than 40-fold in petioles and 15-fold in leaves) and salt (tenfold in leaves). Young leaves and roots contained 97.89 and 91.17 µmol‧g−1 of GSL, whereas less GSL was produced in mature leaves and petioles (38.36 and 44.79 µmol‧g−1, respectively). Similar pattern was observed in the accumulation of GHPs in various plant organs. Notably, despite the non-significant changes in GSL production, abiotic factors treated samples enhanced significantly GHP content. Pearson’s correlation analysis revealed that WjMYRI-1 expression significantly correlated with GSL accumulation and GHP formation, suggesting the primary role of WjMYRI-1-encoding putative protein in GSL degradation. In contrast, WjMYRII expression level showed no correlation with GSL or GHP content, suggesting another physiological role of WjMYRII in stress-induced response.ConclusionsIn conclusions, three potential isogenes (WjMYRI-1, WjMYRI-2, and WjMYRII) encoding for different MYR isoforms in W. japonica were identified. Our results provided new insights related to MYR and GSL metabolism which are important for the implications of wasabi in agriculture, food and pharmaceutical industry. Particularly, WjMYRI-1 may be primarily responsible for GSL degradation, whereas WjMYRII (clade II) may be involved in other regulatory pathways induced by abiotic factors.

Growth conditions trigger genotype-specific metabolic responses that affect the nutritional quality of kale cultivars.

Kales (Brassica oleracea convar acephala) are fast-growing, nutritious leafy vegetables ideal for year-round indoor farming. However, selection of best cultivars for growth under artificial lighting necessitates a deeper understanding of leaf metabolism in different kale types. Here we examined a curly leaved cultivar Half Tall and a lacinato type cultivar Black Magic under moderate growth light (130 µmol photons m-1s-1/22°C) and high light (800 µmol photons m-1s-1/26°C) conditions. These conditions induced genotype-dependent differences in nutritionally important metabolites, especially anthocyanins and glucosinolates (GSLs), in the kale cultivars. In the pale green Half Tall, growth under high light conditions did not induce changes in either pigmentation or total GSL content. In contrast, the purple pigmentation of Black Magic intensified due to increased anthocyanin accumulation. Black Magic showed reduced amounts of indole GSLs and increased amounts of aliphatic GSLs under high light conditions, with notable cultivar-specific adjustments in individual GSL species. Correlation analysis of metabolite profiles suggested cultivar-specific metabolic interplay between serine biosynthesis and the production of indole GSLs. RNA sequencing identified candidate genes encoding metabolic enzymes and regulatory components behind anthocyanin and GSL biosynthesis. These findings improve the understanding of leaf metabolism and its effects on the nutritional quality of kale cultivars.

Selenocysteine methyltransferase SMT catalyzed the synthesis of Se-methylselenocysteine to regulate the accumulation of glucosinolates and sulforaphane in broccoli

Selenocysteine methyltransferase (SMT) is a key enzyme involved in the Se metabolism pathway, and it is responsible for the catalysis of Se-methylselenocysteine (SeMSC) compound formation. Previous studies showed that selenium treatment activated SMT expression and promoted the accumulation of glucosinolates (GSLs) and sulforaphane, but the roles and functional mechanisms of SMT in mediating GSLs and sulforaphane synthesis remain unclear. In this study, we identified the BoSMT gene in broccoli and uncovered its roles in mediating GSLs biosynthesis. Transgenic assays revealed that BoSMT is involved in SeMSC biosynthesis in broccoli. More importantly, the contents of GSLs and sulforaphane were significantly increased in the BoSMT-overexpressing broccoli lines but decreased in the knockdown lines, suggesting that BoSMT played a positive role in regulating GSLs and sulforaphane synthesis. Further evidence indicated that BoSMT-mediated overaccumulation of GSLs and sulforaphane might be due to the increase in the endogenous SeMSC content. Compared with the mock (water) treatment, selenite-induced significantly increases of the SeMSC content in the BoSMT-knockdown plants partially compensated the phenotype of GSLs and sulforaphane loss. Compared with the mock treatment, exogenous SeMSC treatment significantly increased the contents of GSL and sulforaphane and activated GSL synthesis-related gene expression, suggesting that SeMSC acted as a positive regulator for GSL and sulforaphane production. Our findings provided novel insights into selenium-mediated GSLs and sulforaphane accumulation. The genetic manipulation of BoSMT might be a useful strategy for improving the dietary nutritional values of broccoli.

Molecular constraints on tolerance-resistance trade-offs: Is there a cost?

Plants possess myriad defenses against their herbivores, including constitutive and inducible chemical compounds and regrowth strategies known as tolerance. Recent studies have shown that plant tolerance and resistance are positively associated given they are co-localized in the same molecular pathway, the oxidative pentose phosphate pathway. However, given that both defensive strategies utilize carbon skeletons from a shared resource pool in the oxidative pentose phosphate pathway there are likely costs in maintaining both resistance-tolerance strategies. Here we investigate fitness costs in maintaining both strategies by utilizing a double knockout of cyp79B2 and cyp79B3, key enzymes in the biosynthetic process of indole glucosinolates, which convert tryptophan to indole-3-acetaldoxime (IAOx) and is further used to produce indole glucosinolates. These mutant plants are devoid of any indole glucosinolates thus reducing plant resistance. Results show that knocking out indole glucosinolate production and thus one of the resistance pathways leads to an approximate 94% increase in fitness compensation shifting the undercompensating wild-type Columbia-0 to an overcompensating genotype following damage. We discuss the potential mechanistic basis for the observed patterns.

Altered methionine metabolism impacts phenylpropanoid production and plant development in Arabidopsis thaliana.

Phenylpropanoids are specialized metabolites derived from phenylalanine. Glucosinolates are defense compounds derived mainly from methionine and tryptophan in Arabidopsis. It was previously shown that the phenylpropanoid pathway and glucosinolate production are metabolically linked. The accumulation of indole-3-acetaldoxime (IAOx), the precursor of tryptophan-derived glucosinolates, represses phenylpropanoid biosynthesis through accelerated degradation of phenylalanine ammonia lyase (PAL). As PAL functions at the entry point of the phenylpropanoid pathway, which produces indispensable specialized metabolites such as lignin, aldoxime-mediated phenylpropanoid repression is detrimental to plant survival. Although methionine-derived glucosinolates in Arabidopsis are abundant, any impact of aliphatic aldoximes (AAOx) derived from aliphatic amino acids such as methionine on phenylpropanoid production remains unclear. Here, we investigate the impact of AAOx accumulation on phenylpropanoid production using Arabidopsis aldoxime mutants, ref2 and ref5. REF2 and REF5 metabolize aldoximes to respective nitrile oxides redundantly, but with different substrate specificities. ref2 and ref5 mutants have decreased phenylpropanoid contents due to the accumulation of aldoximes. As REF2 and REF5 have high substrate specificity toward AAOx and IAOx, respectively, it was assumed that ref2 accumulates AAOx, not IAOx. Our study indicates that ref2 accumulates both AAOx and IAOx. Removing IAOx partially restored phenylpropanoid content in ref2, but not to the wild-type level. However, when AAOx biosynthesis was silenced, phenylpropanoid production and PAL activity in ref2 were completely restored, suggesting an inhibitory effect of AAOx on phenylpropanoid production. Further feeding studies revealed that the abnormal growth phenotype commonly observed in Arabidopsis mutants lacking AAOx production is a consequence of methionine accumulation.

The mysterious non-arbuscular mycorrhizal status of Brassicaceae species.

The Brassicaceae family is unique in not fostering functional symbiosis with arbuscular mycorrhiza (AM). The family is also special in possessing glucosinolates, a class of secondary metabolites predominantly functioning for plant defence. We have reviewed what effect the glucosinolates of this non-symbiotic host have on AM or vice versa. Isothiocyanates, the toxic degradation product of the glucosinolates, particularly the indolic and benzenic glucosinolates, are known to be involved in the inhibition of AM. Interestingly, AM colonization enhances glucosinolate production in two AM-host in the Brassicales family- Moringa oleifera and Tropaeolum spp. PHOSPHATE STARVATION RESPONSE 1 (PHR1), a central transcription factor that controls phosphate starvation response also activates the glucosinolate biosynthesis in AM non-host Arabidopsis thaliana. Recently, the advances in whole-genome sequencing, enabling extensive ecological microbiome studies have helped unravel the Brassicaceae microbiome, identifying new mutualists that compensate for the loss of AM symbiosis, and reporting cues for some influence of glucosinolates on the microbiome structure. We advocate that glucosinolate is an important candidate in determining the mycorrhizal status of Brassicaceae and has played a major role in its symbiosis-defence trade-off. We also identify key open questions in this area that remain to be addressed in the future.

The effect of chitosan and β-cyclodextrin on glucosinolate biosynthesis in Brassica rapa ssp. pekinensis (Chinese cabbage) shoot culture

Chitosan is a polycationic polysaccharide derived from chitin, and β-cyclodextrin is a type of macrocyclic oligosaccharide linked by α-1,4 glycosidic bonds. These compounds are recognized as effective elicitors in the biosynthesis of secondary metabolites in plants. These elicitors were studied to assess the growth of shoots and the synthesis of glucosinolates (GSLs) from elicited shoots in Chinese cabbage under controlled in vitro conditions for the first time. Chitosan at 150 mg L−1 supplemented in the optimized shoot induction recovered maximum quantities of total GSLs (7.344 μmol g−1 DW) at the end of 5th week of culture duration, followed by β-cyclodextrin (15 mg L−1) with the synthesis of GSLs (6.379 μmol g−1 DW) at the end of 4th week of culture. The application of chitosan completely deteriorated the growth of shoots, whereas β-cyclodextrin did not affect and even increased the growth of shoots (4.66 g DW). Upon elicitation, the individual got GSLs contents exhibited various fold changes (1.78-to-23.5-fold). Real-time PCR analysis of essential GSLs biosynthesis genes like MAM1, ST5b, AOP2, FMOGS-OX1, CYP83B1, CYP81F2, ST5a, and CYP81F4 revealed substantial higher expression upon elicitation. This present study would provide a steady production of GSLs in Chinese cabbage shoots with the influence of carbohydrate-based elicitors for pharmaceutical or food companies in the future.

The rhizosphere microbiome and host plant glucosinolates exhibit feedback cycles in Brassica rapa.

The rhizosphere microbiome influences many aspects of plant fitness, including production of secondary compounds and defence against insect herbivores. Plants also modulate the composition of the microbial community in the rhizosphere via secretion of root exudates. We tested both the effect of the rhizosphere microbiome on plant traits, and host plant effects on rhizosphere microbes using recombinant inbred lines (RILs) of Brassica rapa that differ in production of glucosinolates (GLS), secondary metabolites that contribute to defence against insect herbivores. First, we investigated the effect of genetic variation in GLS production on the composition of the rhizosphere microbiome. Using a Bayesian Dirichlet-multinomial regression model (DMBVS), we identified both negative and positive associations between bacteria from six genera and the concentration of five GLS compounds produced in plant roots. Additionally, we tested the effects of microbial inoculation (an intact vs. disrupted soil microbiome) on GLS production and insect damage in these RILs. We found a significant microbial treatment × genotype interaction, in which total GLS was higher in the intact relative to the disrupted microbiome treatment in some RILs. However, despite differences in GLS production between microbial treatments, we observed no difference in insect damage between treatments. Together, these results provide evidence for a full feedback cycle of plant-microbe interactions mediated by GLS; that is, GLS compounds produced by the host plant "feed-down" to influence rhizosphere microbial community and rhizosphere microbes "feed-up" to influence GLS production.

Understanding the Complex Functional Interplay between Glucosinolates and Cyanogenic Glycosides in Carica papaya

Glucosinolates (GSLs) and cyanogenic glycosides (CGs) fulfil functions in plant defence and have been reported to be anticancer agents. Generally, GSL-containing plants do not produce CG, and vice versa, CG-containing plants do not synthesise GSLs. However, the production of both GSL and CG compounds was observed in Carica papaya. Additionally, several studies found both GSL glucotropaeolin and CG prunasin in papaya leaves. The advancement of genome technologies can be explored to elucidate the gene functions and other molecular discoveries in plants that might relate to GSLs and CGs. This review aims to discuss the complex interplay of the rare events whereby these two compounds (GSL and CG) co-occur in a bifurcation pathway in papaya. To our knowledge, this is the first review that highlights novel GSL and CG genes in papaya. Furthermore, species-specific pathways in papaya are also discussed and comprehensively described. The transcription factors involved in regulating GSL and CG biosynthesis pathways are also discussed, accompanied by relevant bioinformatic approaches that can help discover potential regulatory genes that control the production of prunasin and glucotropaeolin in papaya.

Transcriptome analysis of the pulp of citrus fruitlets suggests that domestication enhanced growth processes and reduced chemical defenses increasing palatability

To identify key traits brought about by citrus domestication, we have analyzed the transcriptomes of the pulp of developing fruitlets of inedible wild Ichang papeda (Citrus ichangensis), acidic Sun Chu Sha Kat mandarin (C. reticulata) and three palatable segregants of a cross between commercial Clementine (C. x clementina) and W. Murcott (C. x reticulata) mandarins, two pummelo/mandarin admixtures of worldwide distribution. RNA-seq comparison between the wild citrus and the ancestral sour mandarin identified 7267 differentially expressed genes, out of which 2342 were mapped to 117 KEGG pathways. From the remaining genes, a set of 2832 genes was functionally annotated and grouped into 45 user-defined categories. The data suggest that domestication promoted fundamental growth processes to the detriment of the production of chemical defenses, namely, alkaloids, terpenoids, phenylpropanoids, flavonoids, glucosinolates and cyanogenic glucosides. In the papeda, the generation of energy to support a more active secondary metabolism appears to be dependent upon upregulation of glycolysis, fatty acid degradation, Calvin cycle, oxidative phosphorylation, and ATP-citrate lyase and GABA pathways. In the acidic mandarin, downregulation of cytosolic citrate degradation was concomitant with vacuolar citrate accumulation. These changes affected nitrogen and carbon allocation in both species leading to major differences in organoleptic properties since the reduction of unpleasant secondary metabolites increases palatability while acidity reduces acceptability. The comparison between the segregants and the acidic mandarin identified 357 transcripts characterized by the occurrence in the three segregants of additional downregulation of secondary metabolites and basic structural cell wall components. The segregants also showed upregulation of genes involved in the synthesis of methyl anthranilate and furaneol, key substances of pleasant fruity aroma and flavor, and of sugar transporters relevant for sugar accumulation. Transcriptome and qPCR analysis in developing and ripe fruit of a set of genes previously associated with citric acid accumulation, demonstrated that lower acidity is linked to downregulation of these regulatory genes in the segregants. The results suggest that the transition of inedible papeda to sour mandarin implicated drastic gene expression reprograming of pivotal pathways of the primary and secondary metabolism, while palatable mandarins evolved through progressive refining of palatability properties, especially acidity.

MYC transcription factors coordinate tryptophan-dependent defence responses and compromise seed yield in Arabidopsis.

Summary Robust plant immunity negatively affects other fitness traits, including growth and seed production. Jasmonate (JA) confers broad‐spectrum protection against plant consumers by stimulating the degradation of JASMONATE ZIM‐DOMAIN (JAZ) proteins, which in turn relieves repression on transcription factors (TFs) coincident with reduced growth and fecundity. The molecular mechanisms underlying JA‐mediated decreases in fitness remain largely unknown.To assess the contribution of MYC TFs to growth and reproductive fitness at high levels of defence, we mutated three MYC genes in a JAZ‐deficient mutant (jazD) of Arabidopsis thaliana that exhibits strong defence and low seed yield.Genetic epistasis analysis showed that de‐repression of MYC TFs in jazD not only conferred strong resistance to insect herbivory but also reduced shoot and root growth, fruit size and seed yield. We also provided evidence that the JAZ–MYC module coordinates the supply of tryptophan with the production of indole glucosinolates and the proliferation of endoplasmic reticulum bodies that metabolise glucosinolates through the action of β‐glucosidases.Our results establish MYCs as major regulators of growth‐ and reproductive–defence trade‐offs and further indicate that these factors coordinate tryptophan availability with the production of amino acid‐derived defence compounds.

Enhancement of Glucosinolate Formation in Broccoli Sprouts by Hydrogen Peroxide Treatment

Broccoli sprouts are known as a rich source of health-beneficial phytonutrients: glucosinolates and phenolic compounds. The production of phytonutrients can be stimulated by elicitors that activate the plant stress response. The aim of this study was enhancing the nutritional value of broccoli sprouts using hydrogen peroxide (H2O2) as an elicitor. Daily spraying with H2O2 (500–1000 mM) enhanced the accumulation of glucosinolates, doubling their content in the cotyledons of 16/8 h photoperiod-grown 7-day sprouts compared to the water-treated controls. The application of H2O2 on dark-grown sprouts showed a smaller extent of glucosinolate stimulation than with light exposure. The treatment affected sprout morphology without reducing their yield. The H2O2-treated sprouts had shorter hypocotyls and roots, negative root tropism and enhanced root branching. The activated glucosinolate production became evident 24 h after the first H2O2 application and continued steadily until harvest. Applying the same treatment to greenhouse-grown wild rocket plants caused scattered leaf bleaching, a certain increase in glucosinolates but decline in phenolics content. The H2O2 treatment of broccoli sprouts caused a 3.5-fold upregulation of APK1, a gene related to sulfur mobilization for glucosinolate synthesis. Comparing the APK1 expression with the competing gene GSH1 using sulfur for antioxidant glutathione production indicated that glutathione synthesis prevailed in the sprouts over the formation of glucosinolates.

Impacts of elicitors on metabolite production and on antioxidant\xa0potential and tyrosinase inhibition in watercress microshoot cultures

The study has proved the stimulating effects of different strategies of treatments with elicitors on the production of glucosinolates (GSLs), flavonoids, polyphenols, saccharides, and photosynthetic pigments in watercress (Nasturtium officinale) microshoot cultures. The study also assessed antioxidant and anti-melanin activities. The following elicitors were tested: ethephon (ETH), methyl jasmonate (MeJA), sodium salicylate (NaSA), and yeast extract (YeE) and were added on day 10 of the growth period. Cultures not treated with the elicitor were used as control. The total GSL content estimations and UHPLC-DAD-MS/MS analyses showed that elicitation influenced the qualitative and quantitative profiles of GSLs. MeJA stimulated the production of gluconasturtiin (68.34 mg/100 g dried weight (DW)) and glucobrassicin (65.95 mg/100 g DW). The elicitation also increased flavonoid accumulation (max. 1131.33 mg/100 g DW, for 100 μM NaSA, collection after 24 h). The elicitors did not boost the total polyphenol content. NaSA at 100 μM increased the production of total chlorophyll a and b (5.7 times after 24 h of treatment), and 50 μM NaSA caused a 6.5 times higher production of carotenoids after 8 days of treatment. The antioxidant potential (assessed with the CUPRAC FRAP and DPPH assays) increased most after 24 h of treatment with 100 μM MeJA. The assessment of anti-melanin activities showed that the microshoot extracts were able to cause inhibition of tyrosinase (max. 27.84% for 1250 µg/mL).Key points• Elicitation stimulated of the metabolite production in N. officinale microshoots.• High production of pro-health glucosinolates and polyphenols was obtained.• N. officinale microshoots have got tyrosinase inhibition potential.• The antioxidant potential of N. officinale microshoots was evaluated.Graphical abstract

Arabidopsis assemble distinct root-associated microbiomes through the synthesis of an array of defense metabolites.

Plant associated microbiomes are known to confer fitness advantages to the host. Understanding how plant factors including biochemical traits influence host associated microbiome assembly could facilitate the development of microbiome-mediated solutions for sustainable plant production. Here, we examined microbial community structures of a set of well-characterized Arabidopsis thaliana mutants disrupted in metabolic pathways for the production of glucosinolates, flavonoids, or a number of defense signalling molecules. A. thaliana lines were grown in a natural soil and maintained under greenhouse conditions for 4 weeks before collection of roots for bacterial and fungal community profiling. We found distinct relative abundances and diversities of bacterial and fungal communities assembled in the individual A. thaliana mutants compared to their parental lines. Bacterial and fungal genera were mostly enriched than depleted in secondary metabolite and defense signaling mutants, except for flavonoid mutations on fungi communities. Bacterial genera Azospirillum and Flavobacterium were significantly enriched in most of the glucosinolate, flavonoid and signalling mutants while the fungal taxa Sporobolomyces and Emericellopsis were enriched in several glucosinolates and signalling mutants. Whilst the present study revealed marked differences in microbiomes of Arabidopsis mutants and their parental lines, it is suggestive that unknown enzymatic and pleiotropic activities of the mutated genes could contribute to the identified host-associated microbiomes. Notwithstanding, this study revealed interesting gene-microbiota links, and thus represents valuable resource data for selecting candidate A. thaliana mutants for analyzing the links between host genetics and the associated microbiome.

The influence of different wavelengths of LED light on the production of glucosinolates and phenolic compounds and the antioxidant potential in in vitro cultures of Nasturtium officinale (watercress)

Cultures of Nasturtium officinale were cultivated in vitro under illumination with different wavelengths of light-emitting diode (LED) light (white LED light—WLED, blue light—B, red light—R, 70% red and 30% blue light—RB, 50% green, 35% red and 15% blue light—RBG, 50% yellow, 35% red and 15% blue light—RBY, 50% far red, 35% red and 15% blue light—RBfR, 50% UV, 35% red and 15% blue light—RBUV, darkness—D), and under white fluorescent light (WF)—control conditions. The study investigated the influence of the applied lighting conditions on biomass growth and the production of glucosinolates, phenolic compounds, as well as photosynthetic pigments, and soluble sugars. The study showed a significant beneficial effect of the RBG light on biomass growth (Gi = 11.81 after 20 days) and the production of glucosinolates. The total glucosinolate content under these conditions increased 5.8 and 1.4 times in comparison with the WF light and D condition, respectively, reaching 237.92 mg 100 g−1 DW. The production of phenolic compounds, sugars, and photosynthetic pigments was comparable to the production under the control conditions. The antioxidant potential of extracts from the cultivated biomass was assessed by the CUPRAC, DPPH, and FRAP assays. Extracts obtained from the biomass of cultures grown under the RBG light had an antioxidant potential similar to that of the control cultures. This is the first report providing evidence of the stimulating effect of light quality on the biomass yield and production of glucosinolates by N. officinale microshoot cultures in vitro.

Precursor-Boosted Production of Metabolites in Nasturtium officinale Microshoots Grown in Plantform Bioreactors, and Antioxidant and Antimicrobial Activities of Biomass Extracts

The study demonstrated the effects of precursor feeding on the production of glucosinolates (GSLs), flavonoids, polyphenols, saccharides, and photosynthetic pigments in Nasturtium officinale microshoot cultures grown in Plantform bioreactors. It also evaluated the antioxidant and antimicrobial activities of extracts. L-phenylalanine (Phe) and L-tryptophan (Trp) as precursors were tested at 0.05, 0.1, 0.5, 1.0, and 3.0 mM. They were added at the beginning (day 0) or on day 10 of the culture. Microshoots were harvested after 20 days. Microshoots treated with 3.0 mM Phe (day 0) had the highest total GSL content (269.20 mg/100 g DW). The qualitative and quantitative profiles of the GSLs (UHPLC-DAD-MS/MS) were influenced by precursor feeding. Phe at 3.0 mM stimulated the best production of 4-methoxyglucobrassicin (149.99 mg/100 g DW) and gluconasturtiin (36.17 mg/100 g DW). Total flavonoids increased to a maximum of 1364.38 mg/100 g DW with 3.0 mM Phe (day 0), and polyphenols to a maximum of 1062.76 mg/100 g DW with 3.0 mM Trp (day 0). The precursors also increased the amounts of p-coumaric and ferulic acids, and rutoside, and generally increased the production of active photosynthetic pigments. Antioxidant potential increased the most with 0.1 mM Phe (day 0) (CUPRAC, FRAP), and with 0.5 mM Trp (day 10) (DPPH). The extracts of microshoots treated with 3.0 mM Phe (day 0) showed the most promising bacteriostatic activity against microaerobic Gram-positive acne strains (MIC 250–500 µg/mL, 20–21 mm inhibition zones). No extract was cytotoxic to normal human fibroblasts over the tested concentration range (up to 250 μg/mL).

Regulation of glucosinolate biosynthesis.

Glucosinolates are secondary defense metabolites produced by plants of the order Brassicales, which includes the model species Arabidopsis and many crop species. In the past 13 years, the regulation of glucosinolate synthesis in plants has been intensively studied, with recent research revealing complex molecular mechanisms that connect glucosinolate production with responses to other central pathways. In this review, we discuss how the regulation of glucosinolate biosynthesis is ecologically relevant for plants, how it is controlled by transcription factors, and how this transcriptional machinery interacts with hormonal, environmental, and epigenetic mechanisms. We present the central players in glucosinolate regulation, MYB and basic helix-loop-helix transcription factors, as well as the plant hormone jasmonate, which together with other hormones and environmental signals allow the coordinated and rapid regulation of glucosinolate genes. Furthermore, we highlight the regulatory connections between glucosinolates, auxin, and sulfur metabolism and discuss emerging insights and open questions on the regulation of glucosinolate biosynthesis.