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Biofortification of staple crops with enhanced levels of pro\u2010vitamin A (<jats:styled-content style=\"fixed-case\">PVA<\/jats:styled-content>) offers a sustainable alternative strategy to both food fortification and supplementation. As a proof of concept, <jats:styled-content style=\"fixed-case\">PVA<\/jats:styled-content>\u2010biofortified transgenic Cavendish bananas were generated and field trialed in Australia with the aim of achieving a target level of 20\u00a0\u03bcg\/g of dry weight (dw) \u03b2\u2010carotene equivalent (\u03b2\u2010<jats:styled-content style=\"fixed-case\">CE<\/jats:styled-content>) in the fruit. Expression of a Fe'i banana\u2010derived phytoene synthase 2a (<jats:italic>MtPsy2a<\/jats:italic>) gene resulted in the generation of lines with <jats:styled-content style=\"fixed-case\">PVA<\/jats:styled-content> levels exceeding the target level with one line reaching 55\u00a0\u03bcg\/g dw \u03b2\u2010<jats:styled-content style=\"fixed-case\">CE<\/jats:styled-content><jats:bold>.<\/jats:bold> Expression of the maize phytoene synthase 1 (<jats:italic>ZmPsy1<\/jats:italic>) gene, used to develop \u2018Golden Rice 2\u2019, also resulted in increased fruit <jats:styled-content style=\"fixed-case\">PVA<\/jats:styled-content> levels although many lines displayed undesirable phenotypes. Constitutive expression of either transgene with the maize polyubiquitin promoter increased <jats:styled-content style=\"fixed-case\">PVA<\/jats:styled-content> accumulation from the earliest stage of fruit development. In contrast, <jats:styled-content style=\"fixed-case\">PVA<\/jats:styled-content> accumulation was restricted to the late stages of fruit development when either the banana 1\u2010aminocyclopropane\u20101\u2010carboxylate oxidase or the expansin 1 promoters were used to drive the same transgenes. Wild\u2010type plants with the longest fruit development time had also the highest fruit <jats:styled-content style=\"fixed-case\">PVA<\/jats:styled-content> concentrations. The results from this study suggest that early activation of the rate\u2010limiting enzyme in the carotenoid biosynthetic pathway and extended fruit maturation time are essential factors to achieve optimal <jats:styled-content style=\"fixed-case\">PVA<\/jats:styled-content> concentrations in banana fruit.<\/jats:p>","DOI":"10.1111\/pbi.12650","type":"journal-article","created":{"date-parts":[[2016,10,14]],"date-time":"2016-10-14T00:25:42Z","timestamp":1476404742000},"page":"520-532","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":181,"title":["Golden bananas in the field: elevated fruit pro\u2010vitamin A from the expression of a single banana transgene"],"prefix":"10.1111","volume":"15","author":[{"given":"Jean\u2010Yves","family":"Paul","sequence":"first","affiliation":[{"name":"Centre for Tropical Crops and Biocommodities Queensland University of Technology Brisbane Qld 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