Vitamin A deficiency is a public health problem in a large number of countries. Biofortification of major staple crops (wheat [Triticum aestivum], rice [Oryza sativa], maize [Zea mays], and potato [Solanum tuberosum]) with β-carotene has the potential to alleviate this nutritional problem. Previously, we engineered transgenic “Golden” potato tubers overexpressing three bacterial genes for β-carotene synthesis (CrtB, CrtI, and CrtY, encoding phytoene synthase, phytoene desaturase, and lycopene β-cyclase, respectively) and accumulating the highest amount of β-carotene in the four aforementioned crops. Here, we report the systematic quantitation of carotenoid metabolites and transcripts in 24 lines carrying six different transgene combinations under the control of the 35S and Patatin (Pat) promoters. Low levels of B-I expression are sufficient for interfering with leaf carotenogenesis, but not for β-carotene accumulation in tubers and calli, which requires high expression levels of all three genes under the control of the Pat promoter. Tubers expressing the B-I transgenes show large perturbations in the transcription of endogenous carotenoid genes, with only minor changes in carotenoid content, while the opposite phenotype (low levels of transcriptional perturbation and high carotenoid levels) is observed in Golden (Y-B-I) tubers. We used hierarchical clustering and pairwise correlation analysis, together with a new method for network correlation analysis, developed for this purpose, to assess the perturbations in transcript and metabolite levels in transgenic leaves and tubers. Through a “guilt-by-profiling” approach, we identified several endogenous genes for carotenoid biosynthesis likely to play a key regulatory role in Golden tubers, which are candidates for manipulations aimed at the further optimization of tuber carotenoid content.

Understanding the relationship between biodiversity and ecosystem functioning (BEF) is crucial to predicting the consequences of ongoing global biodiversity loss. However, what drives BEF relationships in natural ecosystems under globally changing conditions remains poorly understood. To address this knowledge gap, we applied a trait‐based approach to data from coastal dune plant communities distributed along a natural environmental stress gradient. Specifically, we compared the relative importance of below‐ground and above‐ground traits in predicting productivity, decomposition, water regulation, carbon stock and nutrient pools, and tested how these BEF relationships were modulated by environmental stress and the presence of rare species that are typically excluded from experimental systems. Below‐ground traits were just as important as above‐ground traits in driving ecosystem functioning. Moreover, despite having low abundances, rare species positively influenced ecosystem multifunctionality (EMF). However, most biodiversity effects became weaker as environmental stress increased. Our study shows that to understand variation in ecosystem functioning we must consider below‐ground traits as much as above‐ground ones. Moreover, it highlights the importance of conserving rare species for maintaining EMF. However, our findings also suggest that rapid global change could dampen the positive effects of diversity on ecosystem functioning. Read the free Plain Language Summary for this article on the Journal blog.