Dually biofortified cisgenic tomatoes with increased flavonoids and branched-chain amino acids content.

Higher dietary intakes of flavonoids may have a beneficial role in cardiovascular disease prevention. Additionally, supplementation of branched-chain amino acids (BCAAs) in vegan diets can reduce risks associated to their deficiency, particularly in older adults, which can cause loss of skeletal muscle strength and mass. Most plant-derived foods contain only small amounts of BCAAs, and those plants with high levels of flavonoids are not eaten broadly. Here we describe the generation of metabolically engineered cisgenic tomatoes enriched in both flavonoids and BCAAs. In this approach, coding and regulatory DNA elements, all derived from the tomato genome, were combined to obtain a herbicide-resistant version of an acetolactate synthase (mSlALS) gene expressed broadly and a MYB12-like transcription factor (SlMYB12) expressed in a fruit-specific manner. The mSlALS played a dual role, as a selectable marker as well as being key enzyme in BCAA enrichment. The resulting cisgenic tomatoes were highly enriched in Leucine (21-fold compared to wild-type levels), Valine (ninefold) and Isoleucine (threefold) and concomitantly biofortified in several antioxidant flavonoids including kaempferol (64-fold) and quercetin (45-fold). Comprehensive metabolomic and transcriptomic analysis of the biofortified cisgenic tomatoes revealed marked differences to wild type and could serve to evaluate the safety of these biofortified fruits for human consumption.

Fortification and bioaccessibility of saffron apocarotenoids in potato tubers.

Carotenoids are C40 isoprenoids with well-established roles in photosynthesis, pollination, photoprotection, and hormone biosynthesis. The enzymatic or ROS-induced cleavage of carotenoids generates a group of compounds named apocarotenoids, with an increasing interest by virtue of their metabolic, physiological, and ecological activities. Both classes are used industrially in a variety of fields as colorants, supplements, and bio-actives. Crocins and picrocrocin, two saffron apocarotenoids, are examples of high-value pigments utilized in the food, feed, and pharmaceutical industries. In this study, a unique construct was achieved, namely O6, which contains CsCCD2L, UGT74AD1, and UGT709G1 genes responsible for the biosynthesis of saffron apocarotenoids driven by a patatin promoter for the generation of potato tubers producing crocins and picrocrocin. Different tuber potatoes accumulated crocins and picrocrocin ranging from 19.41-360 to 105-800 µg/g DW, respectively, with crocetin, crocin 1 [(crocetin-(ß-D-glucosyl)-ester)] and crocin 2 [(crocetin)-(ß-D-glucosyl)-(ß-D-glucosyl)-ester)] being the main compounds detected. The pattern of carotenoids and apocarotenoids were distinct between wild type and transgenic tubers and were related to changes in the expression of the pathway genes, especially from PSY2, CCD1, and CCD4. In addition, the engineered tubers showed higher antioxidant capacity, up to almost 4-fold more than the wild type, which is a promising sign for the potential health advantages of these lines. In order to better investigate these aspects, different cooking methods were applied, and each process displayed a significant impact on the retention of apocarotenoids. More in detail, the in vitro bioaccessibility of these metabolites was found to be higher in boiled potatoes (97.23%) compared to raw, baked, and fried ones (80.97, 78.96, and 76.18%, respectively). Overall, this work shows that potatoes can be engineered to accumulate saffron apocarotenoids that, when consumed, can potentially offer better health benefits. Moreover, the high bioaccessibility of these compounds revealed that potato is an excellent way to deliver crocins and picrocrocin, while also helping to improve its nutritional value.

Fruit flesh volatile and carotenoid profile analysis within the Cucumis melo L. species reveals unexploited variability for future genetic breeding.

BACKGROUND: Aroma profile and carotenoids content of melon flesh are two important aspects influencing the quality of this fruit that have been characterized using only selected genotypes. However, the extant variability of the whole species remains unknown. RESULTS: A complete view of the volatile/carotenoid profiles of melon flesh was obtained analyzing 71 accessions, representing the whole diversity of the species. Gas chromatography-mass spectrometry and high-performance liquid chromatography were used to analyze 200 volatile compounds and five carotenoids. Genotypes were classified into two main clusters (high/low aroma), but with a large diversity of differential profiles within each cluster, consistent with the ripening behavior, flesh color and proposed evolutionary and breeding history of the different horticultural groups. CONCLUSION: Our results highlight the huge amount of untapped aroma diversity of melon germplasm, especially of non-commercial types. Also, landraces with high nutritional value with regard to carotenoids have been identified. All this knowledge will encourage melon breeding, facilitating the selection of the genetic resources more appropriate to develop cultivars with new aromatic profiles or to minimize the impact of breeding on melon quality. The newly characterized sources provide the basis for further investigations into specific genes/alleles contributing to melon flesh quality. © 2018 Society of Chemical Industry.

Fruit volatile profiles of two citrus hybrids are dramatically different from those of their parents.

Volatile compounds released from the fruit of two hybrid Citrus genotypes (FxCh90 and FxCh77) were compared to those from their parental varieties, Fortune mandarin and Chandler pummelo. A series of 113 compounds were identified, including 31 esters, 23 aldehydes, 20 alcohols, 17 monoterpenoids, and other compounds. The differences in the volatile profile among these four genotypes were essentially quantitative. The most striking result was that the volatile profile of the hybrids was not intermediate between their parents and completely differed from that of Chandler, but came closer to Fortune. This was because 56 of the 113 volatile compounds in the hybrids showed significantly higher or lower levels than in any of the parents. Such transgressive behavior in these hybrids was not observed for other fruit quality traits, such as acidity or soluble solid content. The combination of volatile profiling and chemometrics can be used to select new Citrus genotypes with a distinct volatile profile.