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Generation of transgenic maize with enhanced provitamin A content.

ABSTRACT: Vitamin A deficiency (VAD) affects over 250 million people worldwide and is one of the most prevalent nutritional deficiencies in developing countries, resulting in significant socio-economic losses. Provitamin A carotenoids such as beta-carotene, are derived from plant foods and are a major source of vitamin A for the majority of the world's population. Several years of intense research has resulted in the production of 'Golden Rice 2' which contains sufficiently high levels of provitamin A carotenoids to combat VAD. In this report, the focus is on the generation of transgenic maize with enhanced provitamin A content in their kernels. Overexpression of the bacterial genes crtB (for phytoene synthase) and crtI (for the four desaturation steps of the carotenoid pathway catalysed by phytoene desaturase and zeta-carotene desaturase in plants), under the control of a 'super gamma-zein promoter' for endosperm-specific expression, resulted in an increase of total carotenoids of up to 34-fold with a preferential accumulation of beta-carotene in the maize endosperm. The levels attained approach those estimated to have a significant impact on the nutritional status of target populations in developing countries. The high beta-carotene trait was found to be reproducible over at least four generations. Gene expression analyses suggest that increased accumulation of beta-carotene is due to an up-regulation of the endogenous lycopene beta-cylase. These experiments set the stage for the design of transgenic approaches to generate provitamin A-rich maize that will help alleviate VAD.

SUBMITTER: Aluru M

PROVIDER: S-EPMC2561147 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Provitamin A-enriched Golden Cassava generated through metabolic engineering has reduced dry matter, elevated oil content and enhanced shelf-life

Project description:In this study, β-carotene concentrations in cassava storage roots were enhanced by co-expression of transgenes for deoxyxylulose-5-phosphate synthase (DXS) and bacterial phytoene synthase (crtB), mediated by the patatin type-1 promoter. Storage roots harvested from field-grown plants accumulated carotenoids to ≤50 μg/g DW, a 15- to 20-fold increase relative to roots from non-transgenic plants. Approximately 85-90% of these carotenoids accumulated as all-trans-β-carotene, the most nutritionally efficacious carotenoid. β-carotene-accumulating storage roots displayed delayed onset of post-harvest physiological deterioration, a major constraint limiting utilization of cassava products. Significant metabolite changes were detected in β-carotene enhanced storage roots. Most significantly, an inverse correlation was observed between β-carotene and dry matter contents, with reductions of 50% to 60% of dry matter content in the highest carotenoid accumulating storage roots of different cultivars. Further analysis confirmed concomitant reduction in starch content, and increased levels of total fatty acids, triacylglycerols, soluble sugars, and abscisic acid. Irish potato engineered to co-express DXS and crtB displayed a similar correlation between β-carotene accumulation, reduced dry matter and starch content, and elevated oil and soluble sugars in tubers. Transcriptome analyses revealed reduced expression of starch biosynthetic genes, ADP-glucose pyrophosphorylase genes, in transgenic, carotene-accumulating cassava roots relative to non-transgenic roots. These findings highlight unintended metabolic consequences of provitamin A biofortification of starch-rich organs and point to strategies for redirecting metabolic flux to restore starch production.

2017-06-22 | GSE100319 | GEO

Unravelling the Effect of Provitamin A Enrichment on Agronomic Performance of Tropical Maize Hybrids.

Project description:Maize is consumed in different traditional diets as a source of macro- and micro-nutrients across Africa. Significant investment has thus been made to develop maize with high provitamin A content to complement other interventions for alleviating vitamin A deficiencies. The current breeding focus on increasing β-carotene levels to develop biofortified maize may affect the synthesis of other beneficial carotenoids. The changes in carotenoid profiles, which are commonly affected by environmental factors, may also lead to a trade-off with agronomic performance. The present study was therefore conducted to evaluate provitamin A biofortified maize hybrids across diverse field environments. The results showed that the difference in accumulating provitamin A and other beneficial carotenoids across variable growing environments was mainly regulated by the genetic backgrounds of the hybrids. Many hybrids, accumulating more than 10 µg/g of provitamin A, produced higher grain yields (>3600 kg/ha) than the orange commercial maize hybrid (3051 kg/ha). These hybrids were also competitive, compared to the orange commercial maize hybrid, in accumulating lutein and zeaxanthins. Our study showed that breeding for enhanced provitamin A content had no adverse effect on grain yield in the biofortified hybrids evaluated in the regional trials. Furthermore, the results highlighted the possibility of developing broadly adapted hybrids containing high levels of beneficial carotenoids for commercialization in areas with variable maize growing conditions in Africa.

| S-EPMC8398423 | biostudies-literature

Overexpression of Sucrose Phosphate Synthase Enhanced Sucrose Content and Biomass Production in Transgenic Sugarcane.

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Enhanced virus resistance in transgenic maize expressing a dsRNA-specific endoribonuclease gene from E. coli.

Project description:Maize rough dwarf disease (MRDD), caused by several Fijiviruses in the family Reoviridae, is a global disease that is responsible for substantial yield losses in maize. Although some maize germplasm have low levels of polygenic resistance to MRDD, highly resistant cultivated varieties are not available for agronomic field production in China. In this work, we have generated transgenic maize lines that constitutively express rnc70, a mutant E. coli dsRNA-specific endoribonuclease gene. Transgenic lines were propagated and screened under field conditions for 12 generations. During three years of evaluations, two transgenic lines and their progeny were challenged with Rice black-streaked dwarf virus (RBSDV), the causal agent of MRDD in China, and these plants exhibited reduced levels of disease severity. In two normal years of MRDD abundance, both lines were more resistant than non-transgenic plants. Even in the most serious MRDD year, six out of seven progeny from one line were resistant, whereas non-transgenic plants were highly susceptible. Molecular approaches in the T12 generation revealed that the rnc70 transgene was integrated and expressed stably in transgenic lines. Under artificial conditions permitting heavy virus inoculation, the T12 progeny of two highly resistant lines had a reduced incidence of MRDD and accumulation of RBSDV in infected plants. In addition, we confirmed that the RNC70 protein could bind directly to RBSDV dsRNA in vitro. Overall, our data show that RNC70-mediated resistance in transgenic maize can provide efficient protection against dsRNA virus infection.

| S-EPMC3621894 | biostudies-literature

Transgene expression and Bt protein content in transgenic Bt maize (MON810) under optimal and stressful environmental conditions.

Project description:Bt protein content in transgenic insect resistant (Bt) maize may vary between tissues within plants and between plants growing under different environmental conditions. However, it is unknown whether and how Bt protein content correlates with transgene expression, and whether this relationship is influenced by stressful environmental conditions. Two Bt maize varieties containing the same transgene cassette (MON 810) were grown under optimal and stressful conditions. Before and during stress exposure, the upper leaves were analysed for transgene expression using quantitative RT-PCR and for Bt content using ELISA. Under optimal conditions there was no significant difference in the transgene expression between the two investigated Bt maize varieties whereas Bt protein content differed significantly. Transgene expression was correlated with Bt protein content in only one of the varieties. Under stressful environmental conditions we found similar transgene expressions as under optimal conditions but Bt content responded differently. These results suggest that Bt content is not only controlled by the transgene expression but is also dependent on the genetic background of the maize variety. Under stressful conditions the concentration of Bt protein is even more difficult to predict.

| S-EPMC4390241 | biostudies-literature

Provitamin A Carotenoids in Grain Reduce Aflatoxin Contamination of Maize While Combating Vitamin A Deficiency.

Project description:Aflatoxin contamination of maize grain and products causes serious health problems for consumers worldwide, and especially in low- and middle-income countries where monitoring and safety standards are inconsistently implemented. Vitamin A deficiency (VAD) also compromises the health of millions of maize consumers in several regions of the world including large parts of sub-Saharan Africa. We investigated whether provitamin A (proVA) enriched maize can simultaneously contribute to alleviate both of these health concerns. We studied aflatoxin accumulation in grain of 120 maize hybrids formed by crossing 3 Aspergillus flavus resistant and three susceptible lines with 20 orange maize lines with low to high carotenoids concentrations. The hybrids were grown in replicated, artificially-inoculated field trials at five environments. Grain of hybrids with larger concentrations of beta-carotene (BC), beta-cryptoxanthin (BCX) and total proVA had significantly less aflatoxin contamination than hybrids with lower carotenoids concentrations. Aflatoxin contamination had negative genetic correlation with BCX (-0.28, p < 0.01), BC (-0.18, p < 0.05), and proVA (-0.23, p < 0.05). The relative ease of breeding for increased proVA carotenoid concentrations as compared to breeding for aflatoxin resistance in maize suggests using the former as a component of strategies to combat aflatoxin contamination problems for maize. Our findings indicate that proVA enriched maize can be particularly beneficial where the health burdens of exposure to aflatoxin and prevalence of VAD converge with high rates of maize consumption.

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Generation of transgenic rice with reduced content of major and novel high molecular weight allergens.

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| S-EPMC4884044 | biostudies-literature

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| S-EPMC3883295 | biostudies-literature

Effectiveness of genomic selection for improving provitamin A carotenoid content and associated traits in cassava.

Project description:Global efforts are underway to develop cassava with enhanced levels of provitamin A carotenoids to sustainably meet increasing demands for food and nutrition where the crop is a major staple. Herein, we tested the effectiveness of genomic selection (GS) for rapid improvement of cassava for total carotenoids content and associated traits. We evaluated 632 clones from Uganda's provitamin A cassava breeding pipeline and 648 West African introductions. At harvest, each clone was assessed for level of total carotenoids, dry matter content, and resistance to cassava brown streak disease (CBSD). All clones were genotyped with diversity array technology and imputed to a set of 23,431 single nucleotide polymorphic markers. We assessed predictive ability of four genomic prediction methods in scenarios of cross-validation, across population prediction, and inclusion of quantitative trait loci markers. Cross-validations produced the highest mean prediction ability for total carotenoids content (0.52) and the lowest for CBSD resistance (0.20), with G-BLUP outperforming other models tested. Across population, predictions showed low ability of Ugandan population to predict the performance of West African clones, with the highest predictive ability recorded for total carotenoids content (0.34) and the lowest for CBSD resistance (0.12) using G-BLUP. By incorporating chromosome 1 markers associated with carotenoids content as independent kernel in the G-BLUP model of a cross-validation scenario, prediction ability slightly improved from 0.52 to 0.58. These results reinforce ongoing efforts aimed at integrating GS into cassava breeding and demonstrate the utility of this tool for rapid genetic improvement.

| S-EPMC8496257 | biostudies-literature

Consumer Perceptions and Acceptability of Traditional Dishes Prepared with Provitamin A-Biofortified Maize and Sweet Potato.

Project description:Vitamin A deficiency (VAD) is prevalent in South Africa, particularly among predominantly poor rural communities. Provitamin A (PVA)-biofortified crops could be used to address VAD; however, there are challenges of poor consumer acceptability. This study investigated the effect of replacing white maize and cream-fleshed sweet potato (CFSP) with PVA-biofortified maize and orange-fleshed sweet potato (OFSP), respectively, on consumer acceptability and perceptions of traditional dishes of rural communities in South Africa. Consumer acceptability of PVA-biofortified phutu (a crumbly maize porridge) served with either curried cabbage, chicken or bambara groundnut, separately, and boiled OFSP was evaluated by black South African adults (n = 120) using a five-point facial hedonic scale. Focus group discussions (FGDs) were conducted with 56 subjects recruited from the consumer panel to assess consumer perceptions of the food samples. The majority of the participants rated the composite dishes containing PVA-biofortified phutu as "4 = good" and the acceptability of the composite dishes varied significantly (p < 0.05). Compared to other age groups, the 50-59-year age group showed higher preference for white phutu and chicken curry, whereas the 30-39-year age group showed higher preference for PVA-biofortified phutu and chicken curry. The acceptability of OFSP and CFSP was similar. The study participants showed positive perceptions of the OFSP, as well as PVA-biofortified phutu if served with either curried chicken or cabbage. The findings suggest that PVA-biofortified maize and OFSP can replace white maize and CFSP, respectively, in selected traditional dishes of the rural communities studied to alleviate VAD.

| S-EPMC6682973 | biostudies-literature

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