Biofortification of major crop plants with iron and zinc - achievements and future directions

Biofortification is a long-term strategy of delivering more iron (Fe) and zinc (Zn) to those most in need. Plant breeding programs within the CGIAR and NARS have made major advances in Fe- and Zn-dense variety development and there have been successful releases of new biofortified varieties. Recent research effort has led to a substantial improvement in our knowledge of Fe and Zn homeostasis and gene regulation, resulting in the identification of candidate genes for marker assisted selection. International cooperation between the agricultural and nutrition community has been strengthened, with numerous implementation and partnership strategies developed and employed over the years. The evidence on the effectiveness of Fe and Zn biofortified crops is slowly building up and the results are encouraging. Biofortification continues to be scaled out and further work is required to reach the general aim of eradicating the hidden hunger of Fe and Zn deficiency in the world’s population and ensuring nutritional security

Calcium Biofortification of Crops–Challenges and Projected Benefits

Despite Calcium (Ca) being an essential nutrient for humans, deficiency of Ca is becoming an ensuing public health problem worldwide. Breeding staple crops with higher Ca concentrations is a sustainable long-term strategy for alleviating Ca deficiency, and particular criteria for a successful breeding initiative need to be in place. This paper discusses current challenges and projected benefits of Ca-biofortified crops. The most important features of Ca nutrition in plants are presented along with explicit recommendations for additional exploration of this important issue. In order for Ca-biofortified crops to be successfully developed, tested, and effectively implemented in most vulnerable populations, further research is required

The influence of food consumption and socio-economic factors on the relationship between zinc and iron intake and status in a healthy population

Objective: To examine Zn and Fe nutritional status of a healthy population by means of anthropometric, dietary and biochemical measurements and to investigate the relationship of usual Zn and Fe dietary intakes to Zn and Fe status. In addition, to examine the impact of food choices and socio-economic factors on Fe and Zn dietary intakes and status with the aim to identify groups at risk of dietary deficiency and suggest factors that may influence the status of these nutrients. Design: Food consumption was assessed by 24 h recall questionnaires. Twenty biochemical parameters were measured, of which Hb, haematocrit, erythrocyte count and plasma concentrations of Fe and Zn were directly related to Fe and Zn nutrition. The prevalence of study participants with inadequate micronutrient intakes was calculated using the Estimated Average Requirement cut-point method. Setting: Serbia, Europe. Subjects: Apparently healthy adults (25-65 years of age). Results: Mean daily Zn and Fe intakes were 9.1mg and 11.6 mg for males and 7.3 mg and 9.4 mg for females, respectively. Five per cent of the study population had inadequate dietary Fe intake and 15-25% had inadequate Zn intake. Lower Hb concentrations were measured in women with lower Zn intakes. No differences in Fe and Zn intakes and status among various socio-economic groups were observed, except for Fe intake between the low-income and affluent groups. Conclusions: Regular follow-ups are needed to ensure that potential deficiencies of Zn and Fe do get recognized and addressed in a timely manner

Nutrient composition of important fish species in Bangladesh and potential contribution to recommended nutrient intakes

Fish, in Bangladesh where malnutrition remains a significant development challenge, is an irreplaceable animal-source food in the diet of millions. However, existing data on the nutrient composition of fish do not reflect the large diversity available and have focused on only a few select nutrients. The purpose of this study was to fill the gaps in existing data on the nutrient profiles of common fish in Bangladesh by analysing the proximate, vitamin, mineral and fatty acid composition of 55 fish, shrimp and prawn species from inland capture, aquaculture and marine capture fisheries. When comparing species, the composition of nutrients of public health significance was diverse. Iron ranged from 0.34 to 19. mg/100. g, zinc from 0.6 to 4.7. mg/100. g, calcium from 8.6 to 1900. mg/100. g, vitamin A from 0 to 2503. μg/100. g and vitamin B12 from 0.50 to 14. μg/100. g. Several species were rich in essential fatty acids, particularly docosohexaenoic acid in capture fisheries species (86-310. mg/100. g). The potential contribution of each species to recommended nutrient intakes (RNIs) for pregnant and lactating women (PLW) and infants was calculated. Seven species for PLW and six species for infants, all from inland capture, and all typically consumed whole with head and bones, could potentially contribute ≥25% of RNIs for three or more of these nutrients, simultaneously, from a standard portion. This illustrates the diversity in nutrient content of fish species and in particular the rich nutrient composition of small indigenous species, which should guide policy and programmes to improve food and nutrition security in Bangladesh

An accurate, reliable, and universal qPCR method to identify homozygous single insert T-DNA with the example of transgenic rice

Early determination of transgenic plants that are homozygous for a single locus T-DNA insert is highly desirable in most fundamental and applied transgenic research. This study aimed to build on an accurate, rapid, and reliable quantitative real-time PCR (qPCR) method to fast-track the development of multiple homozygous transgenic rice lines in the T1 generation, with low copy number to single T-DNA insert for further analyses. Here, a well-established qPCR protocol, based on the OsSBE4 reference gene and the nos terminator, was optimized in the transgenic Japonica rice cultivar Nipponbare, to distinguish homozygous single-insert plants with 100% accuracy. This method was successfully adapted to transgenic Indica rice plants carrying three different T-DNAs, without any modifications to quickly develop homozygous rice plants in the T1 generation. The accuracy of this qPCR method when applied to transgenic Indica rice approached 100% in 12 putative transgenic lines. Moreover, this protocol also successfully detected homozygous single-locus T-DNA transgenic rice plants with two-transgene T-DNAs, a feature likely to become more popular in future transgenic research. The assay was developed utilizing universal primers targeting common sequence elements of gene cassettes (the nos terminator). This assay could therefore be applied to other transgenic plants carrying the nos terminator. All procedures described here use standardized qPCR reaction conditions and relatively inexpensive dyes, such as SYBR Green, thus the qPCR method could be cost-effective and suitable for lower budget laboratories that are involved in rice transgenic research

Analysis of the anti-cancer effects of cincau extract (Premna oblongifolia merr) and other types of non- digestible fibre using faecal fermentation supernatants and Caco-2 cells as a model of the human colon

© 2017 by the authors. Green cincau (Premna oblongifolia Merr) is an Indonesian food plant with a high dietary fibre content. Research has shown that dietary fibre mixtures may be more beneficial for colorectal cancer prevention than a single dietary fibre type. The aim of this study was to investigate the effects of green cincau extract on short chain fatty acid (SCFA) production in anaerobic batch cultures inoculated with human faecal slurries and to compare these to results obtained using different dietary fibre types (pectin, inulin, and cellulose), singly and in combination. Furthermore, fermentation supernatants (FSs) were evaluated in Caco-2 cells for their effect on cell viability, differentiation, and apoptosis. Cincau increased total SCFA concentration by increasing acetate and propionate, but not butyrate concentration. FSs from all dietary fibre sources, including cincau, reduced Caco-2 cell viability. However, the effects of all FSs on cell viability, cell differentiation, and apoptosis were not simply explainable by their butyrate content. In conclusion, products of fermentation of cincau extracts induced cell death, but further work is required to understand the mechanism of action. This study demonstrates for the first time that this Indonesian traditional source of dietary fibre may be protective against colorectal cancer

Constitutive Overexpression of the OsNAS Gene Family Reveals Single-Gene Strategies for Effective Iron- and Zinc-Biofortification of Rice Endosperm

BACKGROUND: Rice is the primary source of food for billions of people in developing countries, yet the commonly consumed polished grain contains insufficient levels of the key micronutrients iron (Fe), zinc (Zn) and Vitamin A to meet daily dietary requirements. Experts estimate that a rice-based diet should contain 14.5 µg g−1 Fe in endosperm, the main constituent of polished grain, but breeding programs have failed to achieve even half of that value. Transgenic efforts to increase the Fe concentration of rice endosperm include expression of ferritin genes, nicotianamine synthase genes (NAS) or ferritin in conjunction with NAS genes, with results ranging from two-fold increases via single-gene approaches to six-fold increases via multi-gene approaches, yet no approach has reported 14.5 µg g−1 Fe in endosperm. METHODOLOGY/PRINCIPAL FINDINGS: Three populations of rice were generated to constitutively overexpress OsNAS1, OsNAS2 or OsNAS3, respectively. Nicotianamine, Fe and Zn concentrations were significantly increased in unpolished grain of all three of the overexpression populations, relative to controls, with the highest concentrations in the OsNAS2 and OsNAS3 overexpression populations. Selected lines from each population had at least 10 µg g−1 Fe in polished grain and two OsNAS2 overexpression lines had 14 and 19 µg g−1 Fe in polished grain, representing up to four-fold increases in Fe concentration. Two-fold increases of Zn concentration were also observed in the OsNAS2 population. Synchrotron X-ray fluorescence spectroscopy demonstrated that OsNAS2 overexpression leads to significant enrichment of Fe and Zn in phosphorus-free regions of rice endosperm. CONCLUSIONS: The OsNAS genes, particularly OsNAS2, show enormous potential for Fe and Zn biofortification of rice endosperm. The results demonstrate that rice cultivars overexpressing single rice OsNAS genes could provide a sustainable and genetically simple solution to Fe and Zn deficiency disorders affecting billions of people throughout the world.Alexander A. T. Johnson, Bianca Kyriacou, Damien L. Callahan, Lorraine Carruthers, James Stangoulis, Enzo Lombi and Mark Teste

Dietary Zn deficiency, the current situation, and potential solutions

Zinc (Zn) deficiency is a world-wide problem and this review presents an overview of the magnitude of Zn deficiency with a particular emphasis on present global challenges, current recommendations for Zn intake, and factors that affect - dietary requirements. The challenges of monitoring Zn status are clarified together with the discussion of relevant Zn bioaccessibility and bioavailability issues. Modern lifestyle factors that may exacerbate Zn deficiency and new strategies of reducing its effects are presented. Biofortification, as a potentially useful strategy for improving Zn status in sensitive populations, has been discussed. The review proposes potential actions that could deliver promising results both in terms of monitoring dietary and physiological Zn status as well as in alleviating dietary Zn deficiency in affected populations.This is the peer-reviewed version of the article: Knez, M.; Stangoulis, J. C. R. Dietary Zn Deficiency, the Current Situation, and Potential Solutions. Nutrition Research Reviews 2021, 1–44, [https://doi.org/10.1017/S0954422421000342

The Linoleic Acid: Dihomo-gamma-Linolenic Acid Ratio (LA:DGLA)-An Emerging Biomarker of Zn Status

Zinc (Zn) deficiency is a common aliment predicted to affect 17% of the world's population. Zinc is a vital micronutrient used for over 300 enzymatic reactions and multiple biochemical and structural processes in the body. Although whole blood, plasma, and urine zinc decrease in severe zinc deficiency, accurate assessment of zinc status, especially in mild to moderate deficiency, is difficult as studies with these biomarkers are often contradictory and inconsistent. Hence, as suggested by the World Health Organization, sensitive and specific biological markers of zinc status are still needed. In this review, we provide evidence to demonstrate that the LA:DGLA ratio (linoleic acid:dihomo-gamma-linolenic acid ratio) may be a useful additional indicator for assessing Zn status more precisely. However, this biomarker needs to be tested further in order to determine its full potential

Measurement of haem and total iron in fish, shrimp and prawn using ICP-MS: implications for dietary iron intake calculations

Twenty-five species of fish, shrimp and prawn from local markets in Bangladesh were analysed for concentrations of total Fe, haem Fe and non-haem Fe by ICP-MS. Total Fe and non-haem Fe concentrations were measured in nitric acid-digested samples and haem Fe was extracted using acidified 80% acetone for 60 min. Total Fe concentrations ranged from 0.55–14.43 mg/100 g FW, and haem Fe% ranged from 18%–93% of total Fe. Repeat extractions with 80% acetone recovered additional haem Fe, suggesting that previous measurement by this technique may have underestimated haem Fe content. Calculation of Fe balance (summing Fe in acetone extracts and Fe in the residue after haem Fe extraction) was not significantly different from total Fe, indicating the two processes recovered the different forms of Fe with similar effectiveness