M. J. I. Shohag

Biofortification and Bioavailability of Rice Grain Zinc as Affected by Different Forms of Foliar Zinc Fertilization

Background Zinc (Zn) biofortification through foliar Zn application is an attractive strategy to reduce human Zn deficiency. However, little is known about the biofortification efficiency and bioavailability of rice grain from different forms of foliar Zn fertilizers. Methodology/Principal Findings Four different Zn forms were applied as a foliar treatment among three rice cultivars under field trial. Zinc bioavailability was assessed by in vitro digestion/Caco-2 cell model. Foliar Zn fertilization was an effective agronomic practice to promote grain Zn concentration and Zn bioavailability among three rice cultivars, especially, in case of Zn-amino acid and ZnSO4. On average, Zn-amino acid and ZnSO4 increased Zn concentration in polished rice up to 24.04% and 22.47%, respectively. On average, Zn-amino acid and ZnSO4 increased Zn bioavailability in polished rice up to 68.37% and 64.43%, respectively. The effectiveness of foliar applied Zn-amino acid and ZnSO4 were higher than Zn-EDTA and Zn-Citrate on improvement of Zn concentration, and reduction of phytic acid, as a results higher accumulation of bioavailable Zn in polished rice. Moreover, foliar Zn application could maintain grain yield, the protein and minerals (Fe and Ca) quality of the polished rice. Conclusions Foliar application of Zn in rice offers a practical and useful approach to improve bioavailable Zn in polished rice. According to current study, Zn-amino acid and ZnSO4 are recommended as excellent foliar Zn forms to ongoing agronomic biofortification.

Changes of folate and other potential health-promoting phytochemicals in legume seeds as affected by germination.

Folate deficiency associated with low dietary intake is a well-documented public health problem, resulting in serious health and socioeconomic burdens. Therefore, optimization of the germination process of different cultivars of legume seeds in relation to the content and composition of folate, vitamin C, and total phenolics and total antioxidant capacity was carried out to maximize the health-promoting properties. The content and composition of folate, vitamin C, and total phenolic and total antioxidant capacities varied between species, among cultivars, and with germination time. During germination, total folate content was maximum at 815.2 μg/100 g fresh weight in soybean sprout and at 675.4 μg/100 g fresh weight in mungbean sprout on the fourth day, which were equivalent to, respectively, 3.5- and 3.9-fold increases in the seeds content, and total folate content strongly decreased thereafter. 5-CH(3)-H(4)folate was the most abundant folate species in legume sprouts and reached a maximum on the fourth day. Vitamin C was not detected in raw seeds, and its content increased sharply in soybean and mungbean sprouts and reached a maximum at the fourth day of germination (29 and 27.7 mg/100 g fresh weight, respectively). Germination of soybean and mungbean for 4 days provided the largest amount of total folate as well as the more stable species 5-CH(3)-H(4)folate and also brought about large amounts of vitamin C and total phenolics and substantial antioxidant capacities.

Metallothionein 2 (SaMT2) from Sedum alfredii Hance confers increased Cd tolerance and accumulation in yeast and tobacco.

Metallothioneins are cysteine-rich metal-binding proteins. In the present study, SaMT2, a type 2 metallothionein gene, was isolated from Cd/Zn co-hyperaccumulator Sedum alfredii Hance. SaMT2 encodes a putative peptide of 79 amino acid residues including two cysteine-rich domains. The transcript level of SaMT2 was higher in shoots than in roots of S. alfredii, and was significantly induced by Cd and Zn treatments. Yeast expression assay showed SaMT2 significantly enhanced Cd tolerance and accumulation in yeast. Ectopic expression of SaMT2 in tobacco enhanced Cd and Zn tolerance and accumulation in both shoots and roots of the transgenic plants. The transgenic plants had higher antioxidant enzyme activities and accumulated less H2O2 than wild-type plants under Cd and Zn treatment. Thus, SaMT2 could significantly enhance Cd and Zn tolerance and accumulation in transgenic tobacco plants by chelating metals and improving antioxidant system.

Natural variation of folate content and composition in spinach (Spinacia oleracea) germplasm.

Breeding to increase folate levels in edible parts of plants, termed folate biofortification, is an economical approach to fight against folate deficiency in humans, especially in the developing world. Germplasm with elevated folates are a useful genetic source for both breeding and direct use. Spinach is one of the well-know vegetables that contains a relatively high amount of folate. Currently, little is known about how much folate, and their composition varies in different spinach accessions. The aim of this study was to investigate natural variation in the folate content and composition of spinach genotypes grown under controlled environmental conditions. The folate content and composition in 67 spinach accessions were collected from the United States Department of Agriculture (USDA) and Asian Vegetable Research and Development Center (AVRDC) germplasm collections according to their origin, grown under control conditions to screen for natural diversity. Folates were extracted by a monoenzyme treatment and analyzed by a validated liquid chromatography (LC) method. The total folate content ranged from 54.1 to 173.2 μg/100 g of fresh weight, with 3.2-fold variation, and was accession-dependent. Four spinach accessions (PI 499372, NSL 6095, PI 261787, and TOT7337-B) have been identified as enriched folate content over 150 μg/100 g of fresh weight. The folate forms found were H(4)-folate, 5-CH(3)-H(4)-folate, and 5-HCO-H(4)-folate, and 10-CHO-folic acid also varied among different accessions and was responsible for variation in the total folate content. The major folate vitamer was represented by 5-CH(3)-H(4)-folate, which on average accounted for up to 52% of the total folate pool. The large variation in the total folate content and composition in diverse spinach accessions demonstrates the great genetic potential of diverse genotypes to be exploited by plant breeders.

Effect of zinc sulfate fortification in germinated brown rice on seed zinc concentration, bioavailability, and seed germination.

Rice is the staple food for more than half of the worlds population and, hence, the main source of a vital micronutrient, zinc (Zn). Unfortunately, the bioavailability of Zn from rice is very low not only due to low content but also due to the presence of some antinutrients such as phytic acid. We investigated the effect of germination and Zn fortification treatment on Zn bioavailability of brown rice from three widely grown cultivars using the Caco-2 cell model to find a suitable fortification level for producing germinated brown rice. The results of this study showed that Zn content in brown rice increased significantly (p < 0.05) as the external Zn concentrations increased from 25 to 250 mg/L. In contrast, no significant influence (p > 0.05) on germination percentage of rice was observed when the Zn supply was lower than 150 mg/L. Zn fortification during the germination process has a significant impact on the Zn content and finally Zn bioavailability. These findings may result from the lower molar ratio of phytic acid to Zn and higher Zn content in Zn fortified germinated brown rice, leading to more bioavailable Zn. Likewise, a significant difference (p < 0.05) was found among cultivars with respect to the capacity for Zn accumulation and Zn bioavailability; these results might be attributed to the difference in the molar ratio of phytic acid to Zn and the concentration of Zn among the cultivars evaluated. Based on global intake of Zn among the world population, we recommend germinated brown rice fortified with 100 mg/L ZnSO(4) as a suitable concentration to use in the germination process, which contains high Zn concentration and Zn bioavailability. In the current study, the cultivar Bing91185 fortified with Zn through the germination process contained a high amount as well as bioavailable Zn, which was identified as the most promising cultivar for further evaluation to determine its efficiency as an improved source of Zn for target populations.

Effects of Foliar Iron Application on Iron Concentration in Polished Rice Grain and Its Bioavailability

Iron (Fe) deficiency in humans caused by inadequate dietary intake is a global nutritional problem. A glass house pot experiment was conducted to evaluate the effect of foliar FeSO₄ containing applications on concentrations of Fe, Zn, and Fe bioavailability in polished rice among five rice cultivars. The results showed that foliar application of FeSO₄, FeSO₄, plus nicotianamine (NA), and FeSO₄ plus NA with ZnSO₄ increased the grain Fe concentration by 16.97%, 29.9%, and 27.08%, respectively. The grain Fe bioavailability also increased by foliar application of FeSO₄, FeSO₄ plus NA, and FeSO₄ plus NA with ZnSO₄; these represent increases of 12.63%, 20.86%, and 18.75%, respectively. Foliar FeSO₄ containing applications improved the Fe bioavailability and might be attributed to the reduction of phytic acid and the increase of Fe concentration in polished rice. Addition of ZnSO₄ to foliar Fe application increased both Fe and Zn content without altering Fe content and bioavailability. In addition, the cultivar difference in Fe and Zn concentration was observed and may be due to the genetic control of leaf absorption and seed deposition of foliar application. Furthermore, the cultivar difference in Fe bioavailability observed might be attributed to the variation of grain Fe, phytic acid, and total phenolics contents among the five rice cultivars. The results suggested that foliar FeSO₄ containing applications represent a promising agricultural approach to reduce Fe deficiency in countries where polished rice is extensively consumed.

Role of sulfur assimilation pathway in cadmium hyperaccumulation by Sedum alfredii Hance

Sedum alfredii Hance is a promising cadmium (Cd) hyperaccumulating plant recently identified in China. However, the physiological and molecular mechanisms underlying Cd accumulation, which differentiate hyperaccumulating ecotype (HE) from non-hyperaccumulating ecotype (NHE) has not been elucidated yet. A hydroponic experiment was conducted to investigate the role of sulfur assimilation pathway in Cd hyperaccumulation by the S. alfredii Hance, by analyzing gene expression pattern in sulfur assimilation pathway and the concentration of some sulfur containing compounds. The results show that, sulfur assimilation pathway was affected by Cd differently in HE and NHE S. alfredii Hance. The gene expression pattern of sulfur assimilation pathway was regulated differently in HE and NHE plants, especially the nicotianamine synthase (NAS). NAS transcript levels in root of HE was 141-fold higher than NHE, while in shoots of HE only 0.31-fold higher than NHE. In HE roots, NAS expression level was maximum 3171-fold higher than shoots, while in NHE plants roots NAS expression level was maximum 45.3-fold higher than shoots. In HE plant roots, sulfur, cysteine and methionine concentrations increased 30%, 46% and 835% respectively, by Cd treatment, but in NHE plants roots, sulfur concentration increased less than 1%, cysteine and methionine concentrations decreased 78.5% and 13.3% respectively, by Cd. Cd exposure increased glutathione levels by 142% in HE but less than 10% in NHE plant roots.

Characterization of 68Zn uptake, translocation, and accumulation into developing grains and young leaves of high Zn-density rice genotype

Zinc (Zn) is an essential micronutrient for humans, but Zn deficiency has become serious as equally as iron (Fe) and vitamin A deficiencies nowadays. Selection and breeding of high Zn-density crops is a suitable, cost-effective, and sustainable way to improve human health. However, the mechanism of high Zn density in rice grain is not fully understood, especially how Zn transports from soil to grains. Hydroponics experiments were carried out to compare Zn uptake and distribution in two different Zn-density rice genotypes using stable isotope technique. At seedling stage, IR68144 showed higher 68Zn uptake and transport rate to the shoot for the short-term, but no significant difference was observed in both genotypes for the long-term. Zn in xylem sap of IR68144 was consistently higher, and IR68144 exhibited higher Zn absorption ratio than IR64 at sufficient (2.0 μmol/L) or surplus (8.0 μmol/L) Zn supply level. IR64 and IR68144 showed similar patterns of 68Zn accumulation in new leaves at seedling stage and in developing grains at ripening stage, whereas 68Zn in new leaves and grains of IR68144 was consistently higher. These results suggested that a rapid root-to-shoot translocation and enhanced xylem loading capacity may be the crucial processes for high Zn density in rice grains.

Sorption of sulphamethoxazole by the biochars derived from rice straw and alligator flag.

The sorption ability of sulphamethoxazole (SMX) by biochar derived from rice straw (RS) and alligator flag (AF) at 600°C was studied to assess the ability of biochar as adsorbent to remove SMX from aqueous solution. The results indicated that sorption of SMX by biochars was well described using the Langmuir equation (R2 > 0.94), and the maximum sorption parameter (Q) of RS (3650 mg kg−1) was much higher than that of AF (1963 mg kg−1). Temperature had no effect on SMX sorption by biochars, while thermodynamics analysis indicated that the sorption of SMX on both biochars was a spontaneous physical process. The d 250 RS (diameter of RS sieved through 250 µm) and d 150 AF (diameter of AF sieved through 150 µm) showed excellent sorption ability for SMX. The sorption amount of RS was larger than that of AF when pH < 7, whereas, the sorption amount of AF surpassed RS when pH ≥7. The presence of Cu2+ and/or Cd2+ ion at low concentrations (20 mg L−1) significantly (P < 0.05) increased the sorption of SMX on both RS and AF. Our study confirms that biochar derived from the wetland plants could be used as effective adsorbents to remove SMX from aqueous solution.

Effect of ferrous sulfate fortification in germinated brown rice on seed iron concentration and bioavailability

The present study evaluated the effectiveness of germination and iron fortification on iron concentration and bioavailability of brown rice. Iron fortification during germination process with 0.05-2 g/L ferrous sulfate increased the iron concentration in germinated brown rice from 1.1 to 15.6 times than those in raw brown rice. Based on the recommended dietary allowance of iron, maximum germination rate and γ-aminobutyric acid, we recommend the brown rice fortified with 0.25 g/L FeSO(4) as a suitable fortification level to use in germination process. Iron fortification during the germination process has a positive effect on iron concentration and bioavailability. A significant difference was observed among the cultivars in respect to the capacity for iron accumulation and bioavailability. Germination alone could improve in vitro iron solubility, but had no effect on iron bioavailability in Caco-2 cell, the additional fortification process should be combined to get high amount of bioavailable iron from the brown rice.