Contained metabolic engineering in tomatoes by expression of carotenoid biosynthesis genes from the plastid genome

Applications of chloroplast engineering in agriculture and biotechnology will depend critically on success in extending the crop range of chloroplast transformation, and on the feasibility of expressing transgenes in edible organs (such as tubers and fruits), which often are not green and thus are much less active in chloroplast gene expression. We have improved a recently developed chloroplast-transformation system for tomato plants and applied it to engineering one of the central metabolic pathways in fruits: carotenoid biosynthesis. We report that plastid expression of a bacterial lycopene beta-cyclase gene results in herbicide resistance and triggers conversion of lycopene, the main storage carotenoid of tomatoes, to beta-carotene, resulting in fourfold enhanced pro-vitamin A content of the fruits. Our results demonstrate the feasibility of engineering nutritionally important biochemical pathways in non-green plastids by transformation of the chloroplast genome.     

Mechanisms involved in the intestinal absorption of dietary vitamin A and provitamin A carotenoids

Vitamin A is an essential nutrient for humans and is converted to the visual chromophore, 11-cis-retinal, and to the hormone, retinoic acid. Vitamin A in animal-derived foods is found as long chain acyl esters of retinol and these are digested to free fatty acids and retinol before uptake by the intestinal mucosal cell. The retinol is then reesterified to retinyl esters for incorporation into chlylomicrons and absorbed via the lymphatics or effluxed into the portal circulation facilitated by the lipid transporter, ABCA1. Provitamin A carotenoids such as β-carotene are found in plant-derived foods. These and other carotenoids are transported into the mucosal cell by scavenger receptor class B type I (SR-BI). Provitamin A carotenoids are partly converted to retinol by oxygenase and reductase enzymes and the retinol so produced is available for absorption via the two pathways described above. The efficiency of vitamin A and carotenoid intestinal absorption is determined by the regulation of a number of proteins involved in the process. Polymorphisms in genes for these proteins lead to individual variability in the metabolism and transport of vitamin A and carotenoids. This article is part of a Special Issue entitled Retinoid and Lipid Metabolism.     

Lycopene accumulation affects the biosynthesis of some carotenoid-related volatiles independent of ethylene in tomato

For elucidating the regulatory mechanism of ethylene on carotenoid-related volatiles (open chain) compounds and the relationship between lycopene and carotenoid-related volatiles,transgenic tomato fruits in which ACC synthase was suppressed were used.The transgenic tomato fruit showed a significant reduction of lycopene and aroma volatiles with low ethylene production.6-methyl-5-hepten-2-one,6-methyl-5-hepten-2-ol and geranylacetone,which were suspected to be lycopene degradation products,were lower than those in wild type tomato fruits.In order to identify whether lycopene accumulation effects the biosynthesis of some carotenoid-related volatiles independent of ethylene in tomato or not,the capability of both wild type and transgenic tomato fruits discs to convert lycopene into carotenoid-related volatiles was evaluated.The data showed that external lycopene could convert into 6-methyl-5-hepten-2-one and 6-methyl-5-hepten-2-ol in vivo,Indicating that the strong inhibition of ethylene production had no effect on enzymes in the biosynthesis pathway of some carotenoid-related volatiles.Therefore,in ACS-suppression transgenic tomato fruits,the low levels of 6-methyl-5-hepten-2-one,6-methyl-5-hepten-2-ol was due to decreased lycopene accumulation,not ethylene production.Ethylene only affected the accumulation of lycopene,and then indirectly influenceed the level of lycopene-related volatiles.     

Carotenogenesis in Phycomyces

Time course studies of carotenoid production and of mycelial growth in liquid cultures of Phycomyces blakesleeanus wild type [NRRL 1555 (?)], red mutants C9, C10 and C13 and the heterokaryon C2 * C9 are reported. The ratios of the concentrations of lycopene, γ-carotene and β-carotene in the red mutant C13 and in the heterokaryon C2 * C9 during the growth periods were measured. In these strains the concentration of lycopene is close to its final value after 2 days of growth, at a time at which β-carotene is just beginning to be produced. It is suggested that the β-carotene produced late is possibly synthesized via β-zeacarotene.     

Nicotine inhibition of carotenoid cyclization in Cucurbita ficifolia cotyledons

Nicotine inhibits carotenoid cyclization in greening cucurbit cotyledons resulting in the accumulation of acyclic and monocyclic carotenes. Chlorophyll synthesis is also inhibited by the alkaloid.     

Plasma Carotenoid Concentrations before and after Supplementation with Astaxanthin in Middle-Aged and Senior Subjects

A randomized, double-blind human trial was conducted to assess the effect on the plasma carotenoid concentration of 4- or 12-week astaxanthin supplementation (1 or 3 mg/d) of 20 Japanese middle-aged and senior subjects. The plasma carotenoid concentration was significantly higher after the astaxanthin supplementation than that before in both the 1 mg/d (10 subjects) and 3 mg/d (10 subjects) groups.     

Seed coat carotenoids of the cycad genera Dioon,Encephelartos, Macrozamia and Zamia: Evolutionary significance

Seed coat carotenoids of Dioon, Encephelartos, Macrozamia and Zamia with ranges in Africa, the West Indies, Australia and Mexico are simple mixtures which conform to the theoretical primitive angiosperm “magnolian pattern”. Lycopene is the principal pigment of Zamia while Encephelartos, Dioon and Macrozamia coats contain a mixture of the unsubstituted, mono-and di-hydroxy-β-carotenes. The evolutionary significance of these results is discussed.