Systemic biochemical changes in pepper (Capsicum annuum L.) against Rhizoctonia solani by kale (Brassica oleracea var. acephala L.) green manure application.

BACKGROUND: In the search for new alternatives to avoid the problems associated with the use of synthetic chemical fungicides in agriculture, the use of green manure (GrM) could help combat fungal diseases of crops, such as those produced by the necrotrophic pathogen Rhizoctonia solani. In the case of the use of Brassica tissues as GrM, it could have an elicitor capacity for systemic plant resistance. RESULTS: We used kale leaves as a GrM and applied it to pepper plants infected with R. solani. The application of freeze-dried kale tissues to the roots of pepper plants produced a systemic activation of foliar defences via the salicylic acid (SA) and ethylene (ET) pathways, significantly reducing pathogen damage. In addition, this systemic response led to the accumulation of secondary defence metabolites, such as pipecolic acid, hydroxycoumarin and gluconic acid, in leaves. Remarkably, pepper plants treated with lyophilised kale GrM accumulated glucosinolates when infected with R. solani. We also confirmed that autoclaving removed part of the glucobrassicin (85%) and sinigrin (19%) content of the kale tissues. CONCLUSIONS: GrM kale tissues can activate systemic defences in bell pepper against foliar pathogens through SA/ET hormonal pathways, accumulating secondary defence metabolites.

The growth-immunity tradeoff in Brassica oleracea-Xanthomonas campestris pv. campestris pathosystem.

Plant responses against pathogens are influenced by growth immunity tradeoff, which ensure the best use of limited resources. We study how the immobilization of carbon resources and the induction of defensive responses (glucosinolates, phenolic compounds, stomatal closure) can influence the biomass of two Brassica oleracea lines, differing in their resistance, after infection with Xanthomonas campestris pv. campestris. Potentially, the growth immunity tradeoff can be influenced by the activation of all these processes. However, on the contrary of which is normally stated, our results suggest that the loss of biomass caused by pathogen infection is mainly due to the differential accumulation of starch and the immobilization of sugars rather than the reallocation of resources to synthesize secondary metabolites. Moreover, resistance may be related to the effectiveness of the tradeoff, since the resistant line immobilizes resources more efficiently than the susceptible one. Both inbred lines show a different phytohormones profile, which support the hypothesis that they are employing different strategies to defend themselves against the pathogen. This study emphasizes the key role of the primary metabolism in the defence strategies of plants against pathogens.

New Vegetable Brassica Foods: A Promising Source of Bioactive Compounds.

Brassica rapa is grown in northwestern Spain to obtain turnip greens. The tops of the same plants (flower stems with buds) are cut and sell as turnip tops, increasing the value of the crop. This practice could be extended to other brassicas. The objectives of this work are to study the phytochemical potential of tops of coles (Brassica oleracea) and leaf rape (Brassica napus) compared to turnip tops and to compare tops of different coles (cabbage, kale, tronchuda cabbage), which differ in their morphology and use. We evaluated the content of glucosinolates and phenolic compounds and the antioxidant capacity in leaves and tops of the three species. We found that tops had higher amount of glucosinolates than leaves. Phenolic content and antioxidant capacity followed the opposite trend. Therefore, consumption of leaves and tops are complementary, since both type of organs are enriched with different types of compound. Local varieties of kale, curly kale, cabbage and curly leave cabbage are interesting because of their GSLs and phenolic content and antioxidant capacity in both leaves and tops. From the human health perspective, tops of coles and leaf rape are interesting as new crops to include in the diet.

Glucosinolate Induction and Resistance to the Cabbage Moth, Mamestra brassicae, Differs among Kale Genotypes with High and Low Content of Sinigrin and Glucobrassicin.

The cabbage moth, Mamestra brassicae L. (Lepidoptera: Noctuidae), is a generalist insect pest of cruciferous crops. We tested glucosinolate induction by jasmonic acid (JA) and salicylic acid (SA), and by these phytohormones combined with feeding by M. brassicae larvae in four genotypes of kale, Brassica oleracea L. var. acephala (Brassicaceae). The genotypes tested had high glucobrassicin (genotype HGBS), low glucobrassicin (genotype LGBS), high sinigrin (genotype HSIN), and low sinigrin content (genotype LSIN). Application of JA increased indolic and total glucosinolate content in all kale genotypes 1, 3, and 9 days after treatment. For SA-treated plants, glucosinolate induction varied depending on the number of days after treatment and the genotype. Overall, herbivory by M. brassicae accentuated and attenuated the effects of JA and SA, respectively, on plant glucosinolate content. Larvae of M. brassicae gained less weight on leaves from plants treated with JA compared to leaves from control plants and plants treated with SA. In bioassays with leaf discs, a significant reduction of defoliation only occurred in JA-treated plants of the HSIN genotype. This research shows that previous herbivory alters the susceptibility of kale to M. brassicae and that induction of glucosinolates varies among kale genotypes differing in their glucosinolate content.

Sclerotinia sclerotiorum Response to Long Exposure to Glucosinolate Hydrolysis Products by Transcriptomic Approach.

White mold disease, caused by the necrotrophic fungus Sclerotinia sclerotiorum, affects Brassica crops. Brassica crops produce a broad array of compounds, such as glucosinolates, which contribute to the defense against pathogens. From their hydrolysis, several products arise that have antimicrobial activity (GHPs) whose toxicity is structure dependent. S. sclerotiorum may overcome the toxic effect of moderate GHP concentrations after prolonged exposure to their action. Our objective was to identify the molecular mechanism underlying S. sclerotiorum response to long exposure to two chemically diverse GHPs: aliphatic GHP allyl-isothiocyanate (AITC) and indole GHP indol-3-carbinol (I3C). We found that the transcriptomic response is dependent on the type of GHP and on their initial target, involving cell membranes in the case of AITC or DNA in the case of I3C. Response mechanisms include the reorganization of chromatin, mediated by histone chaperones hip4 and cia1, ribosome synthesis controlled by the kinase-phosphatase pair aps1-ppn1, catabolism of proteins, ergosterol synthesis, and induction of detoxification systems. These mechanisms probably help S. sclerotiorum to grow and survive in an environment where GHPs are constantly produced by Brassica plants upon glucosinolate breakdown. IMPORTANCEBrassica species, including important vegetable crops, such as cabbage, cauliflower, or broccoli, or oil crops, such as rapeseed, produce specific chemical compounds useful to protect them against pests and pathogens. One of the most destructive Brassica diseases in temperate areas around the world is Sclerotinia stem rot, caused by the fungus Sclerotinia sclerotiorum. This is a generalist pathogen that causes disease over more than 400 plant species, being a serious threat to economically important crops worldwide, including potato, bean, soybean, and sunflower, among many others. Understanding the mechanisms utilized by pathogens to overcome specific plant defensive compounds can be useful to increase plant resistance. Our study demonstrated that Sclerotinia shows different adaptation mechanisms, including detoxification systems, to grow and survive when plant protective compounds are present.

Changes in Brassica oleracea Leaves Infected With Xanthomonas campestris pv. campestris by Proteomics Analysis.

Understanding plant's response mechanisms against pathogenesis is fundamental for the development of resistant crop varieties and more productive agriculture. In this regard, "omic" approaches are heralded as valuable technologies. In this work, combining isobaric tags for relative and absolute quantification (iTRAQ) technology with mass spectrometry, the proteomes from leaves of Brassica oleracea plants infected with Xanthomonas campestris pv. campestris (Xcc), and control plants at two different post-infection times were compared. Stronger proteomic changes were obtained at 12 days post-infection in comparison with 3 days. The responses observed involved different cell processes, from primary metabolism, such as photosynthesis or photorespiration, to other complex processes such as redox homeostasis, hormone signaling, or defense mechanisms. Most of the proteins decreased in the earlier response were involved in energetic metabolism, whereas later response was characterized by a recovery of primary metabolism. Furthermore, our results indicated that proteolysis machinery and reactive oxygen species (ROS) homeostasis could be key processes during this plant-pathogen interaction. Current data provide new insights into molecular mechanisms that may be involved in defense responses of B. oleracea to Xcc.

Potential of Germination in Selected Conditions to Improve the Nutritional and Bioactive Properties of Moringa (Moringa oleifera L.).

Moringa oleifera L. is greatly appreciated for its high content of phytochemicals. Although most parts of moringa tree have been widely studied, seeds remained scarcely explored. The first goal of this study was to investigate the effectiveness of germination to improve the nutritional composition (proximate composition and levels of vitamins B1 and B2), content of bioactive compounds (glucosinolates, phenolics and γ-aminobutyric acid, GABA) and antioxidant activity of moringa seed. Germination improved protein, fat, fiber, riboflavin, phenolics, some individual glucosinolates (GLS) and GABA contents, as well as the antioxidant potential in moringa sprouts, but the extent of the improvement depended on germination conditions. The second objective of this work was to identify the optimal germination conditions to maximize nutritional and bioactive quality of moringa by applying multi-response optimization (response surface methodology, RSM). RSM models indicated that 28 °C and 24 h were the optimal conditions to enhance the accumulation of riboflavin, phenolics and antioxidant activity of sprouts, while the highest GABA and total GLS contents were observed at 36 °C for 96 h and thiamine achieved the maximum content at 36 °C for 24 h. These results show that moringa sprouts are promising functional foods that might be also used as ingredients for the elaboration of novel foodstuffs.

Plant Genebanks: Present Situation and Proposals for Their Improvement. the Case of the Spanish Network.

Genebanks were created by the middle of the twentieth century to preserve cultivated biodiversity when landraces began to be substituted by modern varieties. This move was generally accepted as a necessary step to safeguard the future. After about 75 years of collecting and maintaining genetic resources, the increasing ability of biotechnology to create new variability brings the roles of genebanks in the present and near future into question. As a continuation of several workshops that started in 2014, staff of some representative genebanks have met to discuss how the Spanish Plant Genetic Resources Network can be improved, identifying the following major shortcomings: lack of efficient coordination in the distribution of species among genebanks; too many genebanks; existence of detected and undetected duplicates; insufficient rate of regeneration; insufficient phenotyping, genotyping, and epiphenotyping; unsatisfactory rate of use by end users; and, insufficient funding. As a considerable increase in public funding is unlikely, we propose some strategies to increase the efficiency of the system. The most urgent tasks are to strengthen the rationalization of the network by establishing a clear hierarchy and functions, to improve the information in the base collection by deep characterization including not only phenotypes but also uses and utilities, to progressively replace the active collections with focused core collections constructed to meet users' needs, to optimize regeneration protocols, to limit new collecting expeditions of Spanish crop wild relatives to those growing in threatened habitats, and to develop user-friendly platforms to access germplasm documentation, including a unified system of descriptors and classification categories. Current advances in biotechnology, and especially those in gene editing will have without doubt an impact on the role of genebanks. However, the high number of genes and gene combinations created by evolution they hold cannot be produced by these techniques at present. So, these reservoirs of variability will continue to be indispensable for the near-medium future while the function of all the genes is unveiled. In turn, biotechnologies and gene editing will allow us to take advantage of the information held in genebanks in a more efficient and fast way, contributing to a better rationalization and functioning.

Effect of Temperature Stress on Antioxidant Defenses in Brassica oleracea.

Brassica oleracea crops are exposed to seasonal changes in temperature because of their biennial life cycle. Extreme temperatures (cold and heat) affect the photosynthetic activity and the yield of cabbage (B. oleracea capitata group) and kale (B. oleracea acephala group). We studied the relationship among antioxidant defenses, photosynthesis, and yield under extreme temperatures in both crops. Under these conditions, the plants increase the antioxidant defenses, responding to an increment in reactive oxygen species (ROS). The accumulation of ROS in chloroplasts decreases the chlorophyll content and provokes photoinhibition that leads to a low fixation of CO2 and loss of dry weight. Low temperatures especially increase the antioxidant defenses and decrease the chlorophyll content compared to the heat conditions. However, dry weight losses are higher when plants are grown under heat than under cold conditions, probably because of the inactivation of Rubisco and/or the associated enzymes. Both crops were more resilient to cold than to heat temperatures, the capitata group being more resistant.

Effects of Brassicaceae Isothiocyanates on Prostate Cancer.

Despite the major progress made in the field of cancer biology, cancer is still one of the leading causes of mortality, and prostate cancer (PCa) is one of the most encountered malignancies among men. The effective management of this disease requires developing better anticancer agents with greater efficacy and fewer side effects. Nature is a large source for the development of chemotherapeutic agents, with more than 50% of current anticancer drugs being of natural origin. Isothiocyanates (ITCs) are degradation products from glucosinolates that are present in members of the family Brassicaceae. Although they are known for a variety of therapeutic effects, including antioxidant, immunostimulatory, anti-inflammatory, antiviral and antibacterial properties, nowadays, cell line and animal studies have additionally indicated the chemopreventive action without causing toxic side effects of ITCs. In this way, they can induce cell cycle arrest, activate apoptosis pathways, increase the sensitivity of resistant PCa to available chemodrugs, modulate epigenetic changes and downregulate activated signaling pathways, resulting in the inhibition of cell proliferation, progression and invasion-metastasis. The present review summarizes the chemopreventive role of ITCs with a particular emphasis on specific molecular targets and epigenetic alterations in in vitro and in vivo cancer animal models.

Identification of antioxidant capacity -related QTLs in Brassica oleracea.

Brassica vegetables possess high levels of antioxidant metabolites associated with beneficial health effects including vitamins, carotenoids, anthocyanins, soluble sugars and phenolics. Until now, no reports have been documented on the genetic basis of the antioxidant activity (AA) in Brassicas and the content of metabolites with AA like phenolics, anthocyanins and carotenoids. For this reason, this study aimed to: (1) study the relationship among different electron transfer (ET) methods for measuring AA, (2) study the relationship between these methods and phenolic, carotenoid and anthocyanin content, and (3) find QTLs of AA measured with ET assays and for phenolic, carotenoid and anthocyanin contents in leaves and flower buds in a DH population of B. oleracea as an early step in order to identify genes related to these traits. Low correlation coefficients among different methods for measuring AA suggest that it is necessary to employ more than one method at the same time. A total of 19 QTLs were detected for all traits. For AA methods, seven QTLs were found in leaves and six QTLs were found in flower buds. Meanwhile, for the content of metabolites with AA, two QTLs were found in leaves and four QTLs were found in flower buds. AA of the mapping population is related to phenolic compounds but also to carotenoid content. Three genomic regions determined variation for more than one ET method measuring AA. After the syntenic analysis with A. thaliana, several candidate genes related to phenylpropanoid biosynthesis are proposed for the QTLs found.

Identification of metabolic QTLs and candidate genes for glucosinolate synthesis in Brassica oleracea leaves, seeds and flower buds.

Glucosinolates are major secondary metabolites found in the Brassicaceae family. These compounds play an essential role in plant defense against biotic and abiotic stresses, but more interestingly they have beneficial effects on human health. We performed a genetic analysis in order to identify the genome regions regulating glucosinolates biosynthesis in a DH mapping population of Brassica oleracea. In order to obtain a general overview of regulation in the whole plant, analyses were performed in the three major organs where glucosinolates are synthesized (leaves, seeds and flower buds). Eighty two significant QTLs were detected, which explained a broad range of variability in terms of individual and total glucosinolate (GSL) content. A meta-analysis rendered eighteen consensus QTLs. Thirteen of them regulated more than one glucosinolate and its content. In spite of the considerable variability of glucosinolate content and profiles across the organ, some of these consensus QTLs were identified in more than one tissue. Consensus QTLs control the GSL content by interacting epistatically in complex networks. Based on in silico analysis within the B. oleracea genome along with synteny with Arabidopsis, we propose seven major candidate loci that regulate GSL biosynthesis in the Brassicaceae family. Three of these loci control the content of aliphatic GSL and four of them control the content of indolic glucosinolates. GSL-ALK plays a central role in determining aliphatic GSL variation directly and by interacting epistatically with other loci, thus suggesting its regulatory effect.

Bottom-up and top-down herbivore regulation mediated by glucosinolates in Brassica oleracea var. acephala.

Quantitative differences in plant defence metabolites, such as glucosinolates, may directly affect herbivore preference and performance, and indirectly affect natural enemy pressure. By assessing insect abundance and leaf damage rate, we studied the responses of insect herbivores to six genotypes of Brassica oleracea var. acephala, selected from the same cultivar for having high or low foliar content of sinigrin, glucoiberin and glucobrassicin. We also investigated whether the natural parasitism rate was affected by glucosinolates. Finally, we assessed the relative importance of plant chemistry (bottom-up control) and natural enemy performance (top-down control) in shaping insect abundance, the ratio of generalist/specialist herbivores and levels of leaf damage. We found that high sinigrin content decreased the abundance of the generalist Mamestra brassicae (Lepidoptera, Noctuidae) and the specialist Plutella xylostella (Lepidoptera, Yponomeutidae), but increased the load of the specialist Eurydema ornatum (Hemiptera, Pentatomidae). Plants with high sinigrin content suffered less leaf injury. The specialist Brevicoryne brassicae (Hemiptera, Aphididae) increased in plants with low glucobrassicin content, whereas the specialists Pieris rapae (Lepidoptera, Pieridae), Aleyrodes brassicae (Hemiptera, Aleyrodidae) and Phyllotreta cruciferae (Coleoptera, Chrysomelidae) were not affected by the plant genotype. Parasitism rates of M. brassicae larvae and E. ornatum eggs were affected by plant genotype. The ratio of generalist/specialist herbivores was positively correlated with parasitism rate. Although both top-down and bottom-up forces were seen to be contributing, the key factor in shaping both herbivore performance and parasitism rate was the glucosinolate concentration, which highlights the impact of bottom-up forces on the trophic cascades in crop habitats.

In vivo and in vitro effects of secondary metabolites against Xanthomonas campestris pv. campestris.

Brassica rapa is a crucifer that is grown worldwide, mainly as a vegetable. The quality of B. rapa crops is highly affected by the disease caused by the bacteria Xanthomonas campestris pv. campestris (Xcc). Glucosinolates and phenolic compounds can confer resistance to Brassica crops against pests and diseases, but few works have been done to evaluate their role in Xcc resistance. The objectives of this work were: (1) to evaluate the in vivo and in vitro antibacterial effect of gluconapin, its isothiocyanate and the methanolic extracts of B. rapa against the type 4 of Xcc, and (2) to test if there is induced resistance mediated by glucosinolates or phenolic compounds in two varieties of B. rapa. Gluconapin and its ITC varieties had an antibacterial effect on the development of Xanthomonas and this effect was strongly dependent on the concentration applied. Methanolic extracts from B. rapa, containing glucosinolates and phenolic compounds, inhibited the growth of these bacteria. Concentration of gluconapin is higher in resistant plants than in the susceptible ones and there is an induction of gluconapin, some flavonoids and sinapic acid 48 to 72 h after inoculation. Gluconapin plays a role in the constitutive resistance to Xcc, while gluconapin, some flavonoids and hydroxycinnamic acids are induced by a Xcc infection but it is not clear if this induction confers resistance to this disease.

Salicylic acid improves root antioxidant defense system and total antioxidant capacities of flax subjected to cadmium.

Cadmium (Cd) disrupts the normal growth and development of plants, depending on their tolerance to this toxic element. The present study was focused on the impacts of exogenous salicylic acid (SA) on the response and regulation of the antioxidant defense system and membrane lipids to 16-day-old flax plantlets under Cd stress. Exposure of flax to high Cd concentrations led to strong inhibition of root growth and enhanced lipid peroxides, membrane permeability, protein oxidation, and hydrogen peroxide (H2O2) production to varying degrees. Concomitantly, activities of the antioxidant enzymes catalase (CAT, EC 1.11.1.6), guaïcol peroxydase (GPX, EC 1.11.1.7), ascorbate peroxydase (APX, EC 1.11.1.11), and superoxide dismutase (SOD, EC 1.15.1.1), and the total antioxidant capacities (2,2'-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity and ferric reducing antioxidant power (FRAP)) were significantly altered by Cd. In contrast, exogenous SA greatly reduced the toxic effects of Cd on the root growth, antioxidant system, and membrane lipid content. The Cd-treated plantlets pre-soaked with SA exhibited less lipid and protein oxidation and membrane alteration, as well as a high level of total antioxidant capacities and increased activities of antioxidant enzymes except of CAT. These results may suggest that SA plays an important role in triggering the root antioxidant system, thereby preventing membrane damage as well as the denaturation of its components.

Mating system of Brassica napus and its relationship with morphological and ecological parameters in northwestern Spain.

Mating systems play a central role in determining population genetic structure and the methods to be used to develop new cultivars and preserve the variability of a crop. A Brassica napus crop called nabicol is grown in northwestern Spain. Knowledge on its mating system is needed in order to manage the germplasm correctly and design breeding strategies. The aims of this work were to study the mating system of nabicol under field conditions and the relationship of different traits with the mating system. We analyzed 2 populations with microsatellites using a multilocus approach, finding that both had a mixed mating system with an outcrossing rate of 30%. This system would allow application of breeding methods for both autogamous and allogamous species in order to improve nabicol populations. Nabicol populations should be multiplied in isolation conditions in the same way as allogamous species in order to avoid contamination and preserve genetic integrity. The relationship of outcrossing rate, phenological, ecological, and morphological traits was studied, but the model explained only a small percentage of the variability. None of the traits studied could be used as indirect selection criteria for a type of mating system under the conditions of northwestern Spain. This is the first work that studies in depth the possible causes of the mixed mating system of B. napus, finding that, surprisingly, it is not related to the most obvious factors.

Glucosinolate variation in leaves of Brassica rapa crops.

Total and individual glucosinolate (GSL) content of leaves of vegetable turnip rape (Brassica rapa L. var. rapa) was determined in a set of 45 varieties consisting in early, medium and late types grown at two locations in northwestern Spain. The objectives were to determine the diversity among varieties in GSL content and to relate that variation with earliness and plant habit. Eight GSL were identified, being two aliphatic GSL, gluconapin (84.4 % of the total GSL) and glucobrassicanapin (7.2 % of the total GSL) the most abundant. Indolic and aromatic GSL content were low but also showed significant differences among varieties. Differences in total and individual GSL content were found among varieties, plant habit groups, and earliness groups. Total GSL content ranged from 19 to 37.3 µmol g(-1) dw in early and extra-late groups, respectively, and from 19.5 to 36.3 µmol g(-1) dw for turnips and turnip greens groups, respectively. These differences were consistent to values found for gluconapin content where the turnip group had the highest values (31.8 µmol g(-1) dw) and the turnip top group had the lowest (15.7 µmol g(-1) dw). Two varieties, MBG-BRS0429 and MBG-BRS0550 (from turnip greens and extra-late groups) and MBG-BRS0438 (from turnips and late groups), stood out as they had the highest total GSL content and could be used as a good source of these beneficial bioactive compounds. Elucidation of genetic diversity among crops can provide useful information to assist plant breeders to design improved breeding strategies in order to obtain varieties rich on GSL.

Environmental and genetic effects on yield and secondary metabolite production in Brassica rapa crops.

Twelve Brassica rapa varieties grown, such as turnip green and turnip top, were evaluated in seven environments to determine the environmental and genotypic variables that have an influence on crop production and on the content of glucosinolates and phenolic compounds. Factorial regression analysis showed that, in general, crop production was favored by high temperatures all along the crop cycle. However, the lack of a period of intense cold could be a limiting factor. The metabolite content seems to be regulated by extreme temperatures (daily maximum and minimum temperatures) rather than by average daily temperatures. With regard to genotypic covariables, turnip top production was significantly affected by traits related to the vegetative development and time to flowering. Meanwhile, turnip green production was largely affected by a sinapoyl derivative compound, which is a precursor of cell wall components. Cross-talk between glucosinolate biosynthesis and phenylpropanoid signaling pathways is suggested.

Effect of genotype and environmental conditions on health-promoting compounds in Brassica rapa.

It is well-known that a variety of factors (genetic and environmental) affect the ultimate metabolite levels in brassica vegetables, although there is still little information about the role that genetics and environment play on glucosinolates and phenolic levels. Total glucosinolates were more abundant in turnip tops (26.02 µmol g(-1) dw) than in turnip greens (17.78 µmol g(-1) dw). On the other hand, total phenolic content was found in higher quantities in turnip greens (43.81 µmol g(-1) dw) than in turnip tops (37.53 µmol g(-1) dw). Aliphatic glucosinolates were clearly regulated by genotype; in contrast, the effects of environment and genotype×environment interaction on the indolic glucosinolate and phenolic compounds content appeared to be the main effects of variation. Identification of genotypes with enhanced and stable levels of these compounds would provide a value-added opportunity for marketing this crop with superior health promotion to consumers.