Arabidopsis CDF3 transcription factor increases carbon and nitrogen assimilation and yield in trans-grafted tomato plants.

Grafting in tomato (Solanum lycopersicum L.) has mainly been used to prevent damage by soil-borne pathogens and the negative effects of abiotic stresses, although productivity and fruit quality can also be enhanced using high vigor rootstocks. In the context of a low nutrients input agriculture, the grafting of elite cultivars onto rootstocks displaying higher Nitrogen Use Efficiency (NUE) supports a direct strategy for yield maximization. In this study we assessed the use of plants overexpressing the Arabidopsis (AtCDF3) or tomato (SlCDF3) CDF3 genes, previously reported to increase NUE in tomato, as rootstocks to improve yield in the grafted scion under low N inputs. We found that the AtCDF3 gene induced greater production of sugars and amino acids, which allowed for greater biomass and fruit yield under both sufficient and limiting N supplies. Conversely, no positive impact was found with the SlCDF3 gene. Hormone analyses suggest that gibberellins (GA4), auxin and cytokinins (tZ) might be involved in the AtCDF3 responses to N. The differential responses triggered by the two genes could be related, at least in part, to the mobility of the AtCDF3 transcript through the phloem to the shoot. Consistently, a higher expression of the target genes of the transcription factor, such as glutamine synthase 2 (SlGS2) and GA oxidase 3 (SlGA3ox), involved in amino acid and gibberellin biosynthesis, respectively, was observed in the leaves of this graft combination. Altogether, our results provided further insights into the mode of action of CDF3 genes and their biotechnology potential for transgrafting approaches.

Spanish Melon Landraces: Revealing Useful Diversity by Genomic, Morphological, and Metabolomic Analysis.

Spain is a secondary centre of the diversification of the melon (Cucumis melo L.), with high diversity represented in highly appreciated landraces belonging to the Flexuosus and Ibericus groups. A collection of 47 accessions of Flexuosus, Chate, Piel de Sapo, Tendral, Amarillo, Blanco, and Rochet was analysed using a genotyping-by-sequencing (GBS) approach. A total of 66,971 quality SNPs were identified. Genetic analysis differentiated Ibericus accessions and exotic materials (Ameri, Momordica, Kachri, and Agrestis), while Flexuous accessions shared ancestry between them. Within the Ibericus group, no clear genomic distinction could be identified for the different landraces evaluated, with accessions of different landraces showing high genetic similarity. The morphological characterization confirmed that the external colour and fruit shape had been used as recognition patterns for Spanish melon landraces, but variability within a landrace exists. Differences were found in the sugars and acid and volatile profiles of the materials. Flexuosus and Chate melons at the immature commercial stage accumulated malic acid and low levels of hexoses, while Ibericus melons accumulated high contents of sucrose and citric acid. Specific trends could be identified in the Ibericus landraces. Tendral accumulated low levels of sugars and citric acid and high of malic acid, maintaining higher firmness, Rochet reached higher levels of sugars, and Amarillo tended to lower malic acid contents. Interestingly, high variability was found within landraces for the acidic profile, offering possibilities to alter taste tinges. The main volatile organic compounds (VOCs) in Flexuosus and Chate were aldehydes and alcohols, with clear differences between both groups. In the Ibericus landraces, general trends for VOC accumulation could be identified, but, again, a high level of variation exists. This situation highlights the necessity to develop depuration programs to promote on-farm in situ conservation and, at the same time, offers opportunities to establish new breeding program targets and to take advantage of these sources of variation.

Carotenoid fortification of zucchini fruits using a viral RNA vector.

BACKGROUND: Carotenoids are health-promoting metabolites in livestock and human diets. Some important crops have been genetically modified to increase their content. Although the usefulness of transgenic plants to alleviate nutritional deficiencies is obvious, their social acceptance has been controversial. RESULTS: Here, we demonstrate an alternative biotechnological strategy for carotenoid fortification of edible fruits in which no transgenic DNA is involved. A viral RNA vector derived from zucchini yellow mosaic virus (ZYMV) was modified to express a bacterial phytoene synthase (crtB), and inoculated to zucchini (Cucurbita pepo L.) leaves nurturing pollinated flowers. After the viral vector moved to the developing fruit and expressed crtB, the rind and flesh of the fruits developed yellow-orange rather than green color. Metabolite analyses showed a substantial enrichment in health-promoting carotenoids, such as α- and ß-carotene (provitamin A), lutein and phytoene, in both rind and flesh. CONCLUSION: Although this strategy is perhaps not free from controversy due to the use of genetically modified viral RNA, our work does demonstrate the possibility of metabolically fortifying edible fruits using an approach in which no transgenes are involved.

Optimization of electronic nose drift correction applied to tomato volatile profiling.

E-noses can be routinely used to evaluate the volatile profile of tomato samples once the sensor drift and standardization issues are adequately solved. Short-term drift can be corrected using a strategy based on a multiplicative drift correction procedure coupled with a PLS adaptation of the component correction. It must be performed specifically for each sequence, using all sequence signals data. With this procedure, a drastic reduction of sensor signal %RSD can be obtained, ranging between 91.5 and 99.7% for long sequences and between 75.7 and 98.8% for short sequences. On the other hand, long-term drift can be fixed up using a synthetic reference standard mix (with a representation of main aroma volatiles of the species) to be included in each sequence that would enable sequence standardization. With this integral strategy, a high number of samples can be analyzed in different sequences, with a 94.4% success in the aggrupation of the same materials in PLS-DA two-dimensional graphical representations. Using this graphical interface, e-noses can be used to developed expandable maps of volatile profile similitudes, which will be useful to select the materials that most resemble breeding objectives or to analyze which preharvest and postharvest procedures have a lower impact on the volatile profile, avoiding the costs and sample limitations of gas chromatography.

Grafting Snake Melon [Cucumis melo L. subsp. melo Var. flexuosus (L.) Naudin] in Organic Farming: Effects on Agronomic Performance; Resistance to Pathogens; Sugar, Acid, and VOC Profiles; and Consumer Acceptance.

The performance of snake melon [Cucumis melo var. flexuosus (L.)] in organic farming was studied under high biotic and salt stress conditions. Soilborne diseases (mainly caused by Macrophomina phaseolina and Neocosmospora falciformis), combined with virus incidence [Watermelon mosaic virus (WMV), Zucchini yellow mosaic virus (ZYMV), and Tomato leaf curl New Delhi virus (ToLCNDV)] and Podosphaera xanthii attacks, reduced yield by more than 50%. Snake melon susceptibility to M. phaseolina and Monosporascus cannonballus was proved in pathogenicity tests, while it showed some degree of resistance to Neocosmospora keratoplastica and N. falciformis. On the contrary, salt stress had a minor impact, although a synergic effect was detected: yield losses caused by biotic stress increased dramatically when combined with salt stress. Under biotic stress, grafting onto the melon F1Pat81 and wild Cucumis rootstocks consistently reduced plant mortality in different agroecological conditions, with a better performance compared to classic Cucurbita commercial hybrids. Yield was even improved under saline conditions in grafted plants. A negative effect was detected, though, on consumer acceptability, especially with the use of Cucurbita rootstocks. Cucumis F1Pat81 rootstock minimized this side effect, which was probably related to changes in the profile of sugars, acids, and volatiles. Grafting affected sugars and organic acid contents, with this effect being more accentuated with the use of Cucurbita rootstocks than with Cucumis. In fact, the latter had a higher impact on the volatile organic compound profile than on sugar and acid profile, which may have resulted in a lower effect on consumer perception. The use of Cucumis rootstocks seems to be a strategy to enable organic farming production of snake melon targeted to high-quality markets in order to promote the cultivation of this neglected crop.

The targeted overexpression of SlCDF4 in the fruit enhances tomato size and yield involving gibberellin signalling.

Tomato is one of the most widely cultivated vegetable crops and a model for studying fruit biology. Although several genes involved in the traits of fruit quality, development and size have been identified, little is known about the regulatory genes controlling its growth. In this study, we characterized the role of the tomato SlCDF4 gene in fruit development, a cycling DOF-type transcription factor highly expressed in fruits. The targeted overexpression of SlCDF4 gene in the fruit induced an increased yield based on a higher amount of both water and dry matter accumulated in the fruits. Accordingly, transcript levels of genes involved in water transport and cell division and expansion during the fruit enlargement phase also increased. Furthermore, the larger amount of biomass partitioned to the fruit relied on the greater sink strength of the fruits induced by the increased activity of sucrose-metabolising enzymes. Additionally, our results suggest a positive role of SlCDF4 in the gibberellin-signalling pathway through the modulation of GA4 biosynthesis. Finally, the overexpression of SlCDF4 also promoted changes in the profile of carbon and nitrogen compounds related to fruit quality. Overall, our results unveil SlCDF4 as a new key factor controlling tomato size and composition.

High throughput FT-MIR indirect analysis of sugars and acids in watermelon.

Indirect measurements of taste-related compounds are required when a high number of samples has to be analyzed in a short period of time, with a minimum cost. For this purpose, FT-MIR partial least square (PLS) regression models for the prediction of total soluble solids, sugars and organic acids have been developed using three sample sets including breeding lines and commercial varieties of watermelon. Specific models with excellent performance were obtained only for sugars. Nevertheless, a general model supposed a compromise between the best and worse models and offered %RMSEP values of 11.3%, 11.1% and 11.7% for fructose, glucose and sucrose respectively. The model was applied to the selection of high content samples (selection pressure 20% and 30%) obtaining good sensitivity levels and mean percentile of selected samples close to the expected values (100% sensitivity). The robustness of FT-MIR models was assessed with predictions of external assays, obtaining reasonable performances.

FT-MIR determination of taste-related compounds in tomato: a high throughput phenotyping analysis for selection programs.

BACKGROUND: Tomato taste is defined by the accumulation of sugars and organic acids. Individual analyses of these compounds using high-performance liquid chromatography (HPLC) or capillary zone electrophoresis (CZE) are expensive, time-consuming and are not feasible for large number of samples, justifying the interest of spectroscopic methods such as Fourier-transform mid-infrared (FT-MIR). This work analyzed the performance of FT-MIR models to determine the accumulation of sugars and acids, considering the efficiency of models obtained with different ranges of variation. RESULTS: FT-MIR spectra (five-bounce attenuated total reflectance, ATR) were used to obtain partial least squares (PLS) models to predict sugar and acid contents in specific sample sets representing different varietal types. A general model was also developed, obtaining R2 values for prediction higher than 0.84 for main components (soluble solids content, fructose, glucose, and citric acid). Root mean squared error of prediction (RMSEP) for these components were lower than 15% of the mean contents and lower than 6% of the highest contents. Even more, the model sensitivity and specificity for those variables with a 10% selection pressure was 100%. That means that all samples with the 10% highest content were correctly identified. The model was applied to an external assay and it exhibited, for main components, high sensitivities (> 70%) and specificities (> 96%). RMSEP values for main compounds were lower than 21% and 13% of the mean and maximum content respectively. CONCLUSION: The models obtained confirm the effectiveness of FT-MIR models to select samples with high contents of taste-related compounds, even when the calibration has not been performed within the same assay. © 2019 Society of Chemical Industry.

Influence of controlled deficit irrigation on tomato functional value.

The effect of controlled deficit irrigation (CDI) on the accumulation of carotenoids, polyphenols and l-ascorbic acid was studied in conventional and high lycopene tomato cultivars. Plants were initially irrigated to cover 100%ETc and after the fruit set phase, the dose was reduced to 75% or 50% of ETc. CDI had no significant effect on the accumulation of carotenoids, while it increased the levels of the hydroxycinnamic acids chlorogenic and ferulic acids, the flavonoid rutin and l-ascorbic acid. Nevertheless, there were important interactions and this effect was highly dependent on the year and site of cultivation. Certain growing areas would be more favorable to supply high quality markets, and, fortunately, CDI would maximize polyphenol (100-75%ETc) and l-ascorbic acid (100-50%ETc) in these areas. A combination of the best genotype and growing area with CDI would offer high quality products, preserving a scarce resource: water.

Polyphenol and l-ascorbic acid content in tomato as influenced by high lycopene genotypes and organic farming at different environments.

The accumulation of polyphenols and l-ascorbic acid was evaluated under conventional (integrated pest management, IPM) and organic farming, as means to increase the accumulation of chemoprotective compounds. The effect of genotype was considerably higher than the growing system, in fact it is determining. 'Kalvert', a high-lycopene cultivar, outstood for the accumulation of most polyphenols, though low-carotenoid cultivars with high accumulation were also detected. Organic farming significantly increased the levels of caffeic acid by 20%, but reduced those of ferulic acid and naringenin by 13% and 15% respectively. A strong interaction with the environment was detected: in Navarra the differences were limited, while in Extremadura lower contents of ferulic acid and higher contents of chlorogenic acid and rutin were found in organic farming for certain cultivars. The effect of organic farming on l-ascorbic acid was dependent on cultivar and environment and it only led to an increase in Extremadura by 58%.

Ectopic Expression of CDF3 Genes in Tomato Enhances Biomass Production and Yield under Salinity Stress Conditions.

Cycling Dof Factor (CDF) transcription factors (TFs) are involved in multiple processes related to plant growth and development. A member of this family, CDF3, has recently been linked in Arabidopsis to the regulation of primary metabolism and abiotic stress responses, but its role in crop production under stress is still unknown. In this study, we characterized tomato plants overexpressing the CDF3 genes from Arabidopsis and tomato and analyzed their effects on growth and yield under salinity, additionally gaining deeper insights into the molecular function of these TFs. Our results provide evidence for higher biomass production and yield in the 35S::AtCDF3 and 35S::SlCDF3 plants, likely due to a higher photosynthetic capacity resulting in increased sucrose availability. Transcriptome analysis revealed that CDF3 genes regulate a set of genes involved in redox homeostasis, photosynthesis performance and primary metabolism that lead to enhanced biomass production. Consistently, metabolomic profiling revealed that CDF3 evokes changes in the primary metabolism triggering enhanced nitrogen assimilation, and disclosed that the amount of some protective metabolites including sucrose, GABA and asparagine were higher in vegetative tissues of CDF3 overexpressing plants. Altogether these changes improved performance of 35S::AtCDF3 and 35S::SlCDF3 plants under salinity conditions. Moreover, the overexpression of CDF3 genes modified organic acid and sugar content in fruits, improving variables related to flavor perception and fruit quality. Overall, our results associate the CDF3 TF with a role in the control of growth and C/N metabolism, and highlight that overexpression of CDF3 genes can substantially improve plant yield.

Assessment of biochar and hydrochar as minor to major constituents of growing media for containerized tomato production.

BACKGROUND: Chars are emerging materials as constituents of growth media. However, chars of different origin differ in their characteristics and more studies are needed to ratify them for such a role. The characteristics of coir mixed with 0%, 10%, 25%, 50%, 75%, and 100% (v/v) of two biochars, from forest waste (BCH-FW) and from olive mill waste (BCH-OMW), and one hydrochar, from forest waste (HYD-FW), and their effects on growth, yield and fruit quality of two tomato cultivars (Gransol RZ and Cuarenteno) were assessed. RESULTS: Chars negatively affected plant growth and yield but not fruit quality. The effect was related to the char dose and was larger in HYD-FW and BCH-FW than in BCH-OMW, despite the high salinity of the latter, and more acute in Cuarenteno than in Gransol RZ. The results were discussed on the basis of the large particle size of BCH-FW, which could have caused low nutrient solution retention and, hence, reduced plant nutrient uptake, and the high water-holding capacity, poor aeration and large CO2 emission of HYD-FW, which could lead to root anoxia. CONCLUSION: BCH-OMW can be used at high proportion in media for tomato cultivation. The use of BCH-FW at a high proportion might be taken into consideration after adjusting particle size, yet this needs additional assays. HYD-FW is inadequate for soilless containerized tomato cultivation. © 2017 Society of Chemical Industry.

Fruit quality assessment of watermelons grafted onto citron melon rootstock.

BACKGROUND: The grafting of watermelons (Citrullus lanatus) is a common technique that increases yield under stressful soil conditions. The most common rootstocks for watermelons are Cucurbita hybrids. However, they often have a negative impact on fruit quality. Exploiting novel Citrullus germplasm such as citron melon (Citrullus lanatus var. citroides) is an alternative to avoid these quality problems. RESULTS: Citron melon has been validated as watermelon rootstock, comparing its effects on watermelon quality to those of Cucurbita hybrids. Larger fruits with thicker rinds were observed in fruits from plants grafted onto both citron and Cucurbita rootstocks. The citron melon had no significant effect on flesh sugars or acid profiles compared to non-grafted watermelons, except for an increase in glucose and malic acid content, which also occurred in the Cucurbita rootstocks. The aroma profile of fruits produced on citron melon was similar to that of the non-grafted and self-grafted controls. The citron rootstock did not display the increased levels of (Z)-6-nonen-1-ol (a compound associated with pumpkin-like odors) found in fruits produced with Cucurbita hybrids. CONCLUSION: The low impact of citron melon rootstock on fruit quality, along with the enhanced resistance against nematodes, make the citron a promising alternative to Cucurbita rootstocks. © 2016 Society of Chemical Industry.

Simultaneous determination of main phenolic acids and flavonoids in tomato by micellar electrokinetic capillary electrophoresis.

A methodology for the determination of tomato phenolic acids and flavonoids has been developed combining MEKC and DAD detection. The influence on polyphenol separation of pH and background electrolyte, BGE (borax, acetonitrile, methanol and SDS concentrations), was studied and optimized using response surface methodology and weighted desirability function. Separation of polyphenols was achieved within 20min at 15°C using 11.3mM borax and 11.2mM SDS adjusted to pH 8.5 as BGE. Validation was performed using standards and tomato extracts. Recoveries ranged from 77 to 106%. Acceptable repeatabilities were obtained for peak area (%RSD <3.1% and <3.7%) and migration times (%RSD <0.2% and <1.4%) for intra- and inter-day respectively. Detection limits ranged between 0.8 and 3.8mgkg-1. Five and seven of these polyphenols were determined in samples of tomato and related species. This methodology will be valuable tool in breeding programs, analyzing a large number of samples.

Tomato as a Source of Carotenoids and Polyphenols Targeted to Cancer Prevention.

A diet rich in vegetables has been associated with a reduced risk of many diseases related to aging and modern lifestyle. Over the past several decades, many researches have pointed out the direct relation between the intake of bioactive compounds present in tomato and a reduced risk of suffering different types of cancer. These bioactive constituents comprise phytochemicals such as carotenoids and polyphenols. The direct intake of these chemoprotective molecules seems to show higher efficiencies when they are ingested in its natural biological matrix than when they are ingested isolated or in dietary supplements. Consequently, there is a growing trend for improvement of the contents of these bioactive compounds in foods. The control of growing environment and processing conditions can ensure the maximum potential accumulation or moderate the loss of bioactive compounds, but the best results are obtained developing new varieties via plant breeding. The modification of single steps of metabolic pathways or their regulation via conventional breeding or genetic engineering has offered excellent results in crops such as tomato. In this review, we analyse the potential of tomato as source of the bioactive constituents with cancer-preventive properties and the result of modern breeding programs as a strategy to increase the levels of these compounds in the diet.

Fast simultaneous determination of prominent polyphenols in vegetables and fruits by reversed phase liquid chromatography using a fused-core column.

A reversed-phase high-performance liquid chromatography method with photodiode array detection has been developed enabling the joint determination of 17 prominent flavonoids and phenolic acids in vegetables and fruits. A multi-segmented gradient program using a fused-core column for the separation of several phenolic classes (phenolic acids and flavonoids) has been optimised. The influence of extraction conditions (sample freeze-drying, ultrasound extraction, solvent composition and extraction time) has been also optimised using response surface methodology with tomato samples as a model. Complete recoveries (76-108%) were obtained for the phenolic compounds present in tomato. The developed method provided satisfactory repeatability in terms of peak area (RSD<2.9%) and retention time (RSD<0.2%) both for standards and real samples. Detection limits ranged between 3 and 44µgkg(-1) for the detected polyphenols. This method is recommended for routine analysis of large number of samples typical of production quality systems or plant breeding programs.

The role of traditional varieties of tomato as sources of functional compounds.

BACKGROUND: Traditional varieties of tomato, usually associated with excellent organoleptic quality, are increasingly appreciated in European quality markets. A collection of 126 populations of 16 traditional varieties from the east of Spain (a secondary diversity center for tomato) have been evaluated over 2 years in order to determine their potential value as sources of functional compounds, including ascorbic acid, lycopene, ß-carotene and total phenolic content. RESULTS: Population and population × year interaction significantly affected lycopene and ascorbic acid contents, while year effect was also significant for ß-carotene. Despite finding some global trends in certain varieties concerning their functional value, high levels of variation have been found at the intra-varietal level. Populations with high levels of the compounds analysed have been found, as well as different levels of intra-population and inter-year variation. Maximum mean contents for both years have reached 308 mg kg(-1) ascorbic acid, 130 mg kg(-1) lycopene, 30 mg kg(-1) ß-carotene and 89 mg caffeic acid 100 g(-1) total phenolic contents, though it is difficult to identify accessions with joint high values of the three compounds. CONCLUSION: These results open the possibility to promote traditional materials as sources of functional compounds, thus strengthening their quality niches and consolidating their price premium. Additionally, these materials could also be used in breeding programs for quality.

High efficiency joint CZE determination of sugars and acids in vegetables and fruits.

In this work, an improved CE method for the medium-throughput determination of main organic acids (oxalate, malate, citrate), the amino acid glutamate and the sugars fructose, glucose and sucrose in several food matrices is described. These compounds have been identified as key components in the taste intensity of fruit and vegetable crops. Using a running buffer with 20 mM 2,6-pyridine dicarboxylic acid pH 12.1 and 0.1% hexadimethrine bromide, replacing it every 5 h to avoid pH decrease, and optimizing capillary conditioning between runs with 58 mM SDS during 2 min at 20 psi, it is possible to effectively quantify these compounds while increasing medium throughput repeatability. This procedure resolves problems such as increases in migration time and reduction of resolution between problematic peaks (malate/citrate and fructose/glucose) detected in a previous method. The new procedure even considerably reduced time analysis down to 12 min. Under optimal conditions, a large number of injections (200) could be administered without any disturbances in the same capillary. The reliability of the proposed method was further investigated with several food matrix samples, including tomato, pepper, muskmelon, winter squash, and orange. This method is recommended for routine analysis of large number of samples typical of production quality systems or plant breeding programs.

Evaluation of genotype and environment effects on taste and aroma flavor components of Spanish fresh tomato varieties.

Taste and aroma related compounds have been analyzed in a collection of four traditional varieties and two tomato hybrids, representing a wide variability in fruit shape and color, grown in different environments: screenhouse and open field. Protected cultivation tended to show lower sugar concentration (fructose and glucose) but similar acid contents (citric, malic, and glutamic acids). The decreased levels of sucrose equivalents and the similar ratios of sucrose equivalents to citric or glutamic acid contents indicated that protected cultivation, despite being useful to reduce the incidence of pests and viral diseases, reduces the organoleptic quality. Additionally, it doubles the interaccession variability and increased the level of intra-accession variability. In the case of aroma, the genotypic effect was considerably higher than the environmental component on the 12 main volatiles analyzed. Only hexanal and methyl salicylate were significantly affected by environment, while 10 out of 12 volatiles were affected by the genotype. Biplot analysis showed that, even in considerably different environments, it is possible to identify genotype-dependent main aroma profiles. In the case of 13 background volatiles, the environment showed no significant effects and the genotypic effect was lower, though it is possible to identify genotypic trends in background notes.

Evaluation of the genotype, environment and their interaction on carotenoid and ascorbic acid accumulation in tomato germplasm.

BACKGROUND: Tomatoes are an important source of antioxidants (carotenoids, vitamin C, etc.) owing to their high level of consumption. There is great interest in developing cultivars with increased levels of lycopene, ß-carotene or L-ascorbic acid. There is necessary to survey new sources of variation. In this study, the potential of improvement for each character in tomato breeding programmes, in a single or joint approach, and the nature of genotype (G), environment (E) and G × E interaction effects in the expression of these characters were investigated. RESULTS: The content of lycopene, ß-carotene and ascorbic acid determined was very high in some phenotypes (up to 281, 35 and 346 mg kg(-1) respectively). The important differences in the three environments studied (with some stressing conditions in several situations) had a remarkable influence in the phenotypic expression of the functional characters evaluated. Nevertheless, the major contribution came from the genotypic effect along with a considerable G × E interaction. CONCLUSION: The joint accumulation of lycopene and ß-carotene has a high genetic component. It is possible to select elite genotypes with high content of both carotenoids in tomato breeding programmes but multi-environment trials are recommended. The improvement of ascorbic acid content is more difficult because the interference of uncontrolled factors mask the real genetic potential. Among the accessions evaluated, there are four accessions with an amazing genetic potential for functional properties that can be used as donor parents in tomato breeding programmes or for direct consumption in quality markets.