Genetically engineered rice containing larger amounts of nicotianamine to enhance the antihypertensive effect

Usuda, Kanako; Wada, Yasuaki; Ishimaru, Yasuhiro; Kobayashi, Takanori; Takahashi, Michiko; Nakanishi, Hiromi; Nagato, Yasuo; Mori, Satoshi; Nishizawa, Naoko K.

doi:10.1111/j.1467-7652.2008.00374.x

Cited by 39 publications

(21 citation statements)

References 36 publications

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“…6). Usuda et al (2009) produced transgenic rice that expressed HvNAS1 under the control of an endosperm-specific promoter, OsGluB-1. NA concentration in brown seeds of this transgenic line increased threefold compared to the NT line, and Fe concentration in polished seeds also increased 1.5-fold (data not shown).…”

Section: Discussionmentioning

confidence: 99%

Overexpression of the Barley Nicotianamine Synthase Gene HvNAS1 Increases Iron and Zinc Concentrations in Rice Grains

Masuda

Usuda

Kobayashi

et al. 2009

Rice

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215

171

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In humans, iron (Fe) and zinc (Zn) deficiencies result in major worldwide health problems. Transgenic technologies to produce Fe-and Zn-biofortified rice varieties offer a promising potential solution. Nicotianamine, the precursor of phytosiderophores, chelates Fe 2+ and Zn 2+ and plays an important role in transporting these metals to both vegetative and reproductive organs within the plant. Our objective was to increase Fe and Zn contents in rice grains by overexpressing the barley nicotianamine synthase gene HvNAS1. HvNAS1-overexpressing transgenic rice showed increased HvNAS1 expression and subsequent increases in endogenous nicotianamine and phytosiderophore content in shoots, roots, and seeds. Fe and Zn concentrations in polished T 1 seeds from transgenic plants increased more than three and twofold, respectively; Fe and Zn concentrations also increased in both polished and brown T 2 seeds. These results suggest that the overproduction of nicotianamine enhances the translocation of Fe and Zn into rice grains.

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Section: Discussionmentioning

confidence: 99%

Overexpression of the Barley Nicotianamine Synthase Gene HvNAS1 Increases Iron and Zinc Concentrations in Rice Grains

Masuda

Usuda

Kobayashi

et al. 2009

Rice

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215

171

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“…Overexpression of the HvNAS1 gene from barley doubles concentrations of Fe and Zn in young leaves and seeds of tobacco (12). Transgenic rice seeds expressing HvNAS1 driven by the seed-specific pGluB-1 promoter also show greater amounts of NA (4-fold) and Fe (1.5-fold) (13). Likewise, overexpression of AtNAS1 in tobacco leads to higher Fe contents in the leaves (11,12).…”

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confidence: 98%

Iron fortification of rice seeds through activation of the nicotianamine synthase gene

Lee

Jeon

Lee

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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316

254

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The most widespread dietary problem in the world is mineral deficiency. We used the nicotianamine synthase (NAS) gene to increase mineral contents in rice grains. Nicotianamine (NA) is a chelator of metals and a key component of metal homeostasis. We isolated activation-tagged mutant lines in which expression of a rice NAS gene, OsNAS3, was increased by introducing 35S enhancer elements. Shoots and roots of the OsNAS3 activationtagged plants (OsNAS3-D1) accumulated more Fe and Zn. Seeds from our OsNAS3-D1 plants grown on a paddy field contained elevated amounts of Fe (2.9-fold), Zn (2.2-fold), and Cu (1.7-fold). The NA level was increased 9.6-fold in OsNAS3-D1 seeds. Analysis by size exclusion chromatography coupled with inductively coupled plasma mass spectroscopy showed that WT and Os-NAS3-D1 seeds contained equal amounts of Fe bound to IP6, whereas OsNAS3-D1 had 7-fold more Fe bound to a low molecular mass, which was likely NA. Furthermore, this activation led to increased tolerance to Fe and Zn deficiencies and to excess metal (Zn, Cu, and Ni) toxicities. In contrast, disruption of OsNAS3 caused an opposite phenotype. To test the bioavailability of Fe, we fed anemic mice with either engineered or WT seeds for 4 weeks and measured their concentrations of hemoglobin and hematocrit. Mice fed with engineered seeds recovered to normal levels of hemoglobin and hematocrit within 2 weeks, whereas those that ate WT seeds remained anemic. Our results suggest that an increase in bioavailable mineral content in rice grains can be achieved by enhancing NAS expression. activation tagging ͉ bioavailability ͉ hemoglobin ͉ metal homeostasis

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“…The supernatants were applied to an ion-exchange purification system, evaporated, and filtered for HPLC as described previously. 34) Metal concentration determination.…”

Section: Soybean Transformationmentioning

confidence: 99%

“…[31][32][33] We also reported that the NA concentration in rice seeds expressing HvNAS1 under the control of a glutelin gene promoter increased from 10 to 49.6 μg/g dry weight (DW), which is at least four times the amount in non-transgenic (NT) rice seeds. 34) We also previously reported that large amounts of NA could be produced in yeast by introducing the Arabidopsis NAS1 gene (AtNAS1). 35) Soybean (Glycine max) is an important crop, due to the large amounts of protein and lipid content in its seeds and its use as a food and energy source (American Soybean Association; http://www.soystats.…”

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Enhanced levels of nicotianamine promote iron accumulation and tolerance to calcareous soil in soybean

Nozoye

Kim

Kakei

et al. 2014

Bioscience, Biotechnology, and Biochemistry

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Iron (Fe) is an essential nutrient in both plants and humans. Fe deficiency on calcareous soil with low Fe availability is a major agricultural problem. Nicotianamine (NA) is one of the Fe chelator in plants, which is involved in metal translocation into seeds, and serves as an antihypertensive substance in humans. In this study, soybean plants overexpressing the barley NA synthase 1 (HvNAS1) gene driven by the constitutive CaMV 35S promoter were produced using Agrobacterium-mediated transformation. The transgenic soybean showed no growth defect and grew normally. The NA content of transgenic soybean seeds was up to four-fold greater than that of non-transgenic (NT) soybean seeds. The level of HvNAS1 expression was positively correlated with the amount of NA, and a high concentration of NA was maintained in the seeds in succeeding generations. The Fe concentration was approximately two-fold greater in transgenic soybean seeds than in NT soybean seeds. Furthermore, the transgenic soybeans showed tolerance to low Fe availability in calcareous soil. Our results suggested that increasing the NA content in soybean seeds by the overexpression of HvNAS1 offers potential benefits for both human health and agricultural productivity.

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Genetically engineered rice containing larger amounts of nicotianamine to enhance the antihypertensive effect

Cited by 39 publications

References 36 publications

Overexpression of the Barley Nicotianamine Synthase Gene HvNAS1 Increases Iron and Zinc Concentrations in Rice Grains

Overexpression of the Barley Nicotianamine Synthase Gene HvNAS1 Increases Iron and Zinc Concentrations in Rice Grains

Iron fortification of rice seeds through activation of the nicotianamine synthase gene

Enhanced levels of nicotianamine promote iron accumulation and tolerance to calcareous soil in soybean

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