Metabolic Engineering of Potato Carotenoid Content through Tuber-Specific Overexpression of a Bacterial Mini-Pathway

BACKGROUND: Since the creation of “Golden Rice”, biofortification of plant-derived foods is a promising strategy for the alleviation of nutritional deficiencies. Potato is the most important staple food for mankind after the cereals rice, wheat and maize, and is extremely poor in provitamin A carotenoids. METHODOLOGY: We transformed potato with a mini-pathway of bacterial origin, driving the synthesis of beta-carotene (Provitamin A) from geranylgeranyl diphosphate. Three genes, encoding phytoene synthase (CrtB), phytoene desaturase (CrtI) and lycopene beta-cyclase (CrtY) from Erwinia, under tuber-specific or constitutive promoter control, were used. 86 independent transgenic lines, containing six different promoter/gene combinations, were produced and analyzed. Extensive regulatory effects on the expression of endogenous genes for carotenoid biosynthesis are observed in transgenic lines. Constitutive expression of the CrtY and/or CrtI genes interferes with the establishment of transgenosis and with the accumulation of leaf carotenoids. Expression of all three genes, under tuber-specific promoter control, results in tubers with a deep yellow (“golden”) phenotype without any adverse leaf phenotypes. In these tubers, carotenoids increase approx. 20-fold, to 114 mcg/g dry weight and beta-carotene 3600-fold, to 47 mcg/g dry weight. CONCLUSIONS: This is the highest carotenoid and beta-carotene content reported for biofortified potato as well as for any of the four major staple foods (the next best event being “Golden Rice 2”, with 31 mcg/g dry weight beta-carotene). Assuming a beta-carotene to retinol conversion of 6∶1, this is sufficient to provide 50% of the Recommended Daily Allowance of Vitamin A with 250 gms (fresh weight) of “golden” potatoes

Identification of alleles of carotenoid pathway genes important for zeaxanthin accumulation in potato tubers

We have investigated the genetics and molecular biology of orange flesh colour in potato (Solanum tuberosum L.). To this end the natural diversity in three genes of the carotenoid pathway was assessed by SNP analyses. Association analysis was performed between SNP haplotypes and flesh colour phenotypes in diploid and tetraploid potato genotypes. We observed that among eleven beta-carotene hydroxylase 2 (Chy2) alleles only one dominant allele has a major effect, changing white into yellow flesh colour. In contrast, none of the lycopene epsilon cyclase (Lcye) alleles seemed to have a large effect on flesh colour. Analysis of zeaxanthin epoxidase (Zep) alleles showed that all (diploid) genotypes with orange tuber flesh were homozygous for one specific Zep allele. This Zep allele showed a reduced level of expression. The complete genomic sequence of the recessive Zep allele, including the promoter, was determined, and compared with the sequence of other Zep alleles. The most striking difference was the presence of a non-LTR retrotransposon sequence in intron 1 of the recessive Zep allele, which was absent in all other Zep alleles investigated. We hypothesise that the presence of this large sequence in intron 1 caused the lower expression level, resulting in reduced Zep activity and accumulation of zeaxanthin. Only genotypes combining presence of the dominant Chy2 allele with homozygosity for the recessive Zep allele produced orange-fleshed tubers that accumulated large amounts of zeaxanthin

Computation of the travelling salesman problem by a shrinking blob

The travelling salesman problem (TSP) is a well known and challenging combinatorial optimisation problem. Its computational intractability has attracted a number of heuristic approaches to generate satisfactory, if not optimal, candidate solutions. Some methods take their inspiration from natural systems, extracting the salient features of such systems for use in classical computer algorithms. In this paper we demonstrate a simple unconventional computation method to approximate the Euclidean TSP using a virtual material approach. The morphological adaptation behaviour of the material emerges from the low-level interactions of a population of particles moving within a diffusive lattice. A 'blob' of this material is placed over a set of data points projected into the lattice, representing TSP city locations, and the blob is reduced in size over time. As the blob shrinks it morphologically adapts to the configuration of the cities. The shrinkage process automatically stops when the blob no longer completely covers all cities. By manually tracing the perimeter of the blob a path between cities is elicited corresponding to a TSP tour. Over 10 runs on 20 randomly generated datasets consisting of 20 cities this simple and unguided method found tours with a mean average tour length of 6.41 % longer than the minimum tours computed by a TSP solver (mean best performance was 4.27 % longer and mean worst performance was 9.22 % longer). We examine the insertion mechanism by which the blob constructs a tour, note some properties and limitations of its performance, and discuss the relationship between the blob TSP and proximity graphs which group points on the plane. The method is notable for its simplicity, novelty and the spatially represented mechanical mode of its operation. We discuss similarities between this method and previously suggested models of human performance on the TSP and suggest possibilities for further improvement. © 2013 Springer Science+Business Media Dordrecht