Research Project: Advancing the Nutritional Quality of Staple Food Crops for Improved Intestinal Function and Health

Location: Plant, Soil and Nutrition Research

2020 Annual Report

Objectives
Objective 1: In collaboration with plant breeders, utilize in vitro, in vivo, and analytical techniques to evaluate Fe bioavailability in staple food crops (beans, wheat, maize) for the purpose of developing crops that serve as an improved source of Fe. [NP107, C1, PS1A] Sub-objective 1.A. Collaborate with breeders to develop bean varieties with enhanced Fe bioavailability that can deliver more absorbable Fe and be a highly sustainable source of dietary Fe. Sub-objective 1.B. Characterize the potential genotype x environment (GxE) interactions that influence Fe content and Fe bioavailability in beans; and determine if either content, bioavailability or both can be sustainable approaches to improve Fe nutrition from beans. Sub-objective 1.C. Collaborate with breeders to characterize Fe bioavailability and Fe concentration from the different components of the maize and wheat grain, and search for QTL for enhanced Fe bioavailability and content of these components. Objective 2. Characterize the individual and interactive effects of dietary minerals (Fe and Zn), phytochemicals and prebiotics common to staple foods (chickpea, bean, wheat, lentil) on the intestinal microbiome and intestinal brush border membrane functionality. [NP107, C3, PS3B] Sub-objective 2.A. Evaluate the effect of natural prebiotics and phytochemicals extracted from staple food crops on mineral (Fe, Zn) dietary bioavailability and absorption in vivo (Gallus gallus).

Approach
As iron and zinc deficiencies are affecting approximately 30% of the world’s populations, including about 15% of the US population, the focus of this research plan is to improve iron bioavailability in staple food crops and to further increase their consumer appeal. Further, we aim to characterize the individual and interactive effects of dietary minerals (Fe and Zn), phytochemicals and prebiotics common to staple foods (chickpea, bean, wheat, lentil) on the intestinal microbiome and intestinal brush border membrane functionality. This will be done in order to evaluate the effect of natural prebiotics and phytochemicals extracted from staple food crops on mineral (Fe, Zn) dietary bioavailability and absorption in vivo (Gallus gallus). In addition, as the newly accepted body “organ”, the intestinal microbiome plays a vital role in the functionality, absorption and digestion capabilities of the intestine; hence, we aim to characterize the microbiome response to dietary plant origin bioactive compounds that may contribute to intestinal functionality and overall health. To accomplish the above, we will employ our established screening tools of an iron bioavailability bioassay and a poultry model that reflects human nutrition. In conjunction, we will make use of techniques such as mass spectroscopy, marker assisted molecular breeding, our Zinc Status biomarker, gene expression, microscopy, and state of the art microbial profiling techniques. With this unique combination of tools, we expect to develop staple food crops with enhanced iron nutrition and elucidate factors that can improve zinc nutrition from staple food crops, and thereby contribute to alleviation of two of the leading micronutrient deficiencies in the world. We will also expand knowledge of how the intestinal microbiome is affected by dietary iron, zinc, phytochemicals and prebiotics. Overall, this knowledge will further contribute to food innovations with enhanced nutrition and improve human health both domestically and abroad.

Progress Report
Significant progress has been made on identifying and developing bean varieties that provide enhanced Fe nutrition. In collaboration with bean breeders, varieties of fast cooking yellow beans with high Fe bioavailability have been developed and we are now close to germplasm and varietal release both domestically and abroad. In related work, in collaboration with bean breeders in North Dakota, we have identified commercially produced popular varieties of pinto beans that provide enhanced Fe nutrition. These pinto bean varieties possess the “slow-darkening” trait which is highly valued by growers and consumers. Our initial studies indicate the mechanism for this nutritional benefit is linked to the profile of the polyphenolic compounds present in the seed coat, with enhanced levels of specific polyphenols identified as the promoters of Fe uptake from these bean varieties. Our bean research has also shown that milling beans into a flour enhances the Fe nutrition from certain color classes (ie. yellow and white), and also maintains a higher concentration of the seed coat polyphenols relative to cooked, non-milled beans. Thus, this proprietary ARS milling process enhances Fe nutrition and helps maintain antioxidant benefits from this important staple food crop. For the past 17 years, the common bean is a crop targeted for iron “biofortification” (ie. higher iron concentration) as a strategy to alleviate the high rate of iron (Fe) deficiency anemia in countries of East Africa. Recently, we conducted research that evaluated the Fe concentration of beans in markets of East African countries, with the goal of comparing the Fe concentration and Fe bioavailability of common marketplace bean varieties with those released as “biofortified” varieties. Our study found the biofortified lines to be similar to the common marketplace varieties. In addition, our research finds that identifying varieties with high Fe bioavailability can be a more sustainable and effective approach for delivering more absorbable dietary Fe. This research is significant as it indicates that the bean Fe biofortification approach of breeding for higher Fe concentration provides no additional dietary Fe, and that bean biofortification efforts should be redefined to include an emphasis on traits that promote Fe bioavailability. Our research group is uniquely positioned with the proper tools and collaborations to lead this redefinition of bean Fe biofortification. Progress in related to Fe/Zn biofortified wheat was made. Specifically, with the further characterization and identification of a specific intestinal microbiome profile under various dietary conditions. For example, a recent comparison between birds receiving a standard wheat-based diet relative to subjects receiving a Zn/Fe biofortified wheat-based diet. Results indicated that dietary Zn/Fe content affected the relative abundances of intestinal microbial species. Through metagenomic analysis, we showed that predicted Kyoto Encyclopedia of Genes and Genomes pathways responsible for macro- and micronutrient uptake are significantly depleted under Zn deficiency; along with concomitant decreases in beneficial short chain fatty acids, such depletions may further preclude optimal host Zn availability. We also identified several candidate microbes that may play a significant role in modulating the bioavailability and utilization of dietary Zn/Fe during prolonged deficiency. These results are significant as they provide and characterize a unique and dysbiotic cecal microbiota during Zn/Fe deficiency, and provide evidence for such microbial perturbations as potential effectors of the Zn/Fe deficient phenotype. Further, and by using the red blood cell fatty acid ratio as a novel biomarker for Zn status, we were able to validate its reactiveness to dietary Zn in wheat- based diets. This is an additional demonstration of Zn biofortified wheat to improve Zn status in vivo. Overall, these findings will allow to better understand the potential effects that dietary Zn has on various physiological process and on dietary Zn bioavailability and absorption. One manuscript was published. Progress was made related to the demonstration that plant (fruit, grain, seed) origin bioactive compounds can improve BBM digestion and function. Chia seed, wheat bran and carioca beans extracts, increased villus growth, goblet cell diameters, mineral metabolism related proteins expression, and the abundance of health promoting bacteria. This provides further demonstration of how functional foods effect the functionality of the intestinal BBM, and in the context of nutrient-gene interactions, immune system, prebiotic properties and interactions with microbiome. Three manuscripts were published.

Accomplishments
1. Development of fast-cooking yellow bean with enhanced iron bioavailability. In countries of East Africa, fast-cooking beans are highly desirable due to the scarcity of cooking fuel and iron deficiency anemia. In the U.S., USDA encourages increased bean consumption. Therefore, fast cooking and improved iron nutrition are essential. ARS scientists at Ithaca, New York, collaborating with ARS researchers at East Lansing, Michigan, used traditional breeding to develop varieties of fast-cooking yellow beans that deliver higher levels of absorbable iron. These new bean varieties provide bean growers and bean breeders with the products to reduce iron deficiency anemia.

Review Publications
Golden, C., Vaitla, B., Ravaoliny, L., Vonona, M., Anjaranirina, E., Randriamady, H., Guth, S., Fernald, L., Myers, S., Glahn, R.P. 2019. Seasonal trends of nutrient intake in rainforest communities of northeastern Madagascar. Public Health Nutrition. 12:2200-2209. https://doi.org/10.1017/S1368980019001083.
Tako, E. 2017. Intra amniotic administration of raffinose and stachyose affects the intestinal brush border functionality and alters gut microflora populations. In: McCrory, M., editor. Dietary Fibers and Human Health. Basel, Switzerland: MDPI. p. 48-58. https://doi.org/10.3390/books978-3-03842-582-3.
Tako, E.N., Mahler, G., Olivas, F.M. 2018. ZnO nanoparticles affect nutrient transport in an in vitro model of the small intestine. Food and Chemical Toxicology. https://doi.org/10.1016/j.fct.2018.11.048.
Hou, T., Tako, E.N. 2018. The in ovo feeding administration (gallus gallus)-an emerging in vivo approach to assess bioactive compounds with potential nutritional benefits. Nutrients. 10(4):418. https://doi.org/10.3390/nu10040418.
Knez, M., Stangoulis, J., Glibetic, M., Tako, E. 2018. The linoleic acid: dihomo-y-linolenic acid ratio (LA:DGLA)—an emerging biomarker of Zn status. In: Tako, E. Dietary Zn and Human Health. Basel, Switzerland: MDPI Books. p. 1-12.
Wang, X., Kolba, N.J., Liang, J., Tako, E.N. 2019. Alterations in gut microflora populations and brush border functionality following intra-amniotic administration (Gallus gallus) of wheat bran prebiotics extracts. Food & Function. 1-10. https://doi.org/10.1039/C9FO00836E.
Podder, R., Dellavalle, D., Tyler, R., Glahn, R.P., Tako, E.N., Vandenberg, A. 2018. Relative bioavailability of iron in Bangledeshi traditional meals prepared with iron-fortified lentil dal. Nutrients. 10(3). https://doi.org/10.3390/nu10030354.
Guo, Z., Martucci, N., Liu, Y., Mahler, G., Yoo, E., Tako, E.N. 2018. Silicon dioxide nanoparticle exposure affects small intestine function in an in vitro model. Nanotoxicology. 12(5):485-508. https://doi.org/10.1080/17435390.2018.1463407.
Glahn, R.P., Hart, J.J., Beebe, S., Tako, E.N. 2018. Iron bioavailability studies of the first generation of iron-biofortified beans released in Rwanda. In: Tako, E., editor. Fe deficiency, dietary bioavailability and absorption. Basel, Switzerland: MDPI Books. p. 16-27. https://doi.org/10.3390/books978-3-03897-231-0.
Glahn, R.P., Tako, E.N., Gore, M.A. 2019. The germ fraction inhibits iron bioavailability of maize: identification of an approach to enhance maize nutritional quality via processing and breeding. Nutrients. 11(4):833. https://doi.org/10.3390/nu11040833.
Wakshlag, J., Glahn, R.P., Kim, H., Loftus, J., Gagne, J., Rutzke, M. 2018. Evaluation of selected ultra-trace minerals in commercially available dry dog foods. Veterinary Medicine: Research and Reports. 9:43-51. https://doi.org/10.2147/VMRR.S165890.
Hooper, S., Glahn, R.P., Cichy, K.A. 2019. Single varietal dry bean (Phaseolus vulgaris L.) pastas: Nutritional profile and consumer acceptability. Plant Foods for Human Nutrition. https://doi.org/10.1007/s11130-019-00732-y.
Ying, L., Glahn, R.P., Hebb, R., Rizvi, S. 2017. Physico-chemical properties, phytochemicals and DPPH radical scavenging activity of supercritical fluid extruded lentils. LWT - Food Science and Technology. 89:315-321. https://doi.org/10.1016/j.lwt.2017.10.063.
Wiesinger, J.A., Glahn, R.P., Cichy, K.A., Kolba, N.J., Hart, J.J., Tako, E.N. 2019. An in vivo (Gallus gallus) feeding trial demonstrates the enhanced iron bioavailability properties of the fast cooking Manteca yellow bean (Phaseolus vulgaris L.). Nutrients. 11(8):1768. https://doi.org/10.3390/nu11081768.
Jahan, T., Vandenberg, A., Glahn, R.P., Tyler, R.T., Reaney, M.J., Tar'An, B. 2019. Iron fortification and bioavailability of chickpea (Cicer arietinum L.) seeds and flour. Nutrients. 11(9):2240. https://doi.org/10.3390/nu11092240.
Beasley, J., Johnaon, A., Glahn, R.P., Hart, J.J., Tako, E.N. 2019. Investigation of Nicotianamine and 2’ Deoxymugineic Acid as enhancers of iron bioavailability in Caco-2 cells. Nutrients. 11(7):1502. https://doi.org/10.3390/nu11071502.
Fleige, L., Bhoite, R., Marwaha, A., Glahn, R.P. 2018. Development of a fortified juice beverage: assessment of iron bioavailability using an established caco-2 cell bioassay. Journal of Nutrition & Food Sciences. 6(3):1-7. https://doi.org/10.15226/jnhfs.2018.001133.
Wiesinger, J.A., Cichy, K.A., Tako, E.N., Glahn, R.P. 2018. The fast cooking and enhanced iron bioavailability properties of the manteca yellow bean (Phaseolus vulgaris L.). Nutrients. 10(11):1609. https://doi.org/10.3390/nu10111609.
Reed, S., Knez, M., Uzan, A., Stangoulis, J., Glahn, R.P., Koren, O., Tako, E.N. 2018. Alterations in the gut (Gallus gallus) microbiota following the consumption of zinc biofortified wheat (Triticum aestivum) -based diet. Journal of Agricultural and Food Chemistry. 66(25):6291-6299. https://doi.org/10.1021/acs.jafc.8b01481.
Beasley, J., Bonneau, J., Sanchez-Palacious, J., Moreno-Moyano, L., Callahan, D., Tako, E.N., Glahn, R.P., Lombi, E., Johnson, A. 2019. Metabolic engineering of bread wheat improves grain iron concentration and bioavailability. Plant Biotechnology Journal. 17(8):1514-1526. https://doi.org/10.1111/pbi.13074.
Morais Dias, D., Kolba, N.J., Binyamin, D., Ziv, O., Regini Nutti, M., Stampini Duarte, M., Glahn, R.P., Koren, O., Tako, E.N. 2018. Iron biofortified carioca bean (phaseolus vulgaris l.) —based Brazilian diet delivers more absorbable iron and affects the gut microbiota in vivo (gallus gallus). Nutrients. 10(12):1970. https://doi.org/10.3390/nu10121970.
Dias, D., Kolba, N.J., Nutti, M., Martino, H., Hart, J.J., Ma, M., Lakshmanan, N., Glahn, R.P., Sha, S., Tako, E.N. 2019. Soluble extracts from carioca beans (Phaseolus vulgaris L.) affect the gut microbiota and iron related brush border membrane protein expression in vivo (Gallus gallus). Food Research International. 123:172-180. https://doi.org/10.1016/j.foodres.2019.04.060.