Yanyan Wei

Improved Plant Growth and Zn Accumulation in Grains of Rice (Oryza sativa L.) by Inoculation of Endophytic Microbes Isolated from a Zn Hyperaccumulator, Sedum alfredii H.

This study is to investigate the possibility of zinc (Zn) biofortification in the grains of rice (Oryza sativa L.) by inoculation of endophytic strains isolated from a Zn hyperaccumulator, Sedum alfredii Hance. Five endophytic strains, Burkholderia sp. SaZR4, Burkholderia sp. SaMR10, Sphingomonas sp. SaMR12, Variovorax sp. SaNR1, and Enterobacter sp. SaCS20, isolated from S. alfredii, were inoculated in the roots of Japonica rice Nipponbare under hydroponic condition. Fluorescence images showed that endophytic strains successfully colonized rice roots after 72 h. Improved root morphology and plant growth of rice was observed after inoculation with endophytic strains especially SaMR12 and SaCS20. Under hydroponic conditions, endophytic inoculation with SaMR12 and SaCS20 increased Zn concentration by 44.4% and 51.1% in shoots, and by 73.6% and 83.4% in roots, respectively. Under soil conditions, endophytic inoculation with SaMR12 and SaCS20 resulted in an increase of grain yields and elevated Zn concentrations by 20.3% and 21.9% in brown rice and by 13.7% and 11.2% in polished rice, respectively. After inoculation of SaMR12 and SaCS20, rhizosphere soils of rice plants contained higher concentration of DTPA-Zn by 10.4% and 20.6%, respectively. In situ micro-X-ray fluorescence mapping of Zn confirmed the elevated Zn content in the rhizosphere zone of rice treated with SaMR12 as compared with the control. The above results suggested that endophytic microbes isolated from S. alfredii could successfully colonize rice roots, resulting in improved root morphology and plant growth, increased Zn bioavailability in rhizosphere soils, and elevated grain yields and Zn densities in grains.

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.

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.

Improved yield and Zn accumulation for rice grain by Zn fertilization and optimized water management

Zinc (Zn) deficiency and water scarcity are major challenges in rice (Oryza sativa L.) under an intensive rice production system. This study aims to investigate the impact of water-saving management and different Zn fertilization source (ZnSO4 and Zn-EDTA) regimes on grain yield and Zn accumulation in rice grain. Different water managements, continuous flooding (CF), and alternate wetting and drying (AWD) were applied during the rice growing season. Compared with CF, the AWD regime significantly increased grain yield and Zn concentrations in both brown rice and polished rice. Grain yield of genotypes (Nipponbare and Jiaxing27), on the average, was increased by 11.4%, and grain Zn concentration by 3.9% when compared with those under a CF regime. Zn fertilization significantly increased Zn density in polished rice, with a more pronounced effect of ZnSO4 being observed as compared with Zn-EDTA, especially under an AWD regime. Decreased phytic acid content and molar ratio of phytic acid to Zn were also noted in rice grains with Zn fertilization. The above results demonstrated that water management of AWD combined with ZnSO4 fertilization was an effective agricultural practice to elevate grain yield and increase Zn accumulation and bioavailability in rice grains.概要研究目的在生产高度集约化体系下, 研究节水灌溉措施及锌肥对水稻产量及锌积累量的影响。创新要点目前关于干湿交替能否对水稻增产仍存在争议, 对锌含量及生物有效性也鲜有报道。 因此, 针对这些问题, 本文系统地研究影响水稻产量及品质的两大限制性因素, 即锌肥和灌溉方式对稻米锌生物有效性的影响。研究方法采用盆钵试验模拟大田锌肥及水分管理方式, 两种基因型、 两种锌肥种类、 两种水分管理, 三个重复控制误差。 利用电感耦合等离子体质谱(ICP-MS)测定微量元素锌含量, 以精米中植酸含量表征锌生物有效性。重要结论干湿交替在增加了稻米产量、 锌含量及积累量方面效果显著。 干湿交替结合土施硫酸锌肥可以作为水稻生产体系中获得高产、 高锌、 高生物有效性的农艺措施。

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.

Iron concentration, bioavailability, and nutritional quality of polished rice affected by different forms of foliar iron fertilizer.

The present study compared the effects of four different forms of foliar iron (Fe) fertilizers on Fe concentration, bioavailability and nutritional quality of polished rice. The results showed that foliar fertilisation at the anthesis stage was an effective way to promote Fe concentration and bioavailability of polished rice, especially in case of DTPA-Fe. Compared to the control, foliar application of DTPA-Fe increased sulphur concentration and the nutrition promoter cysteine content, whereas decreased phosphorus concentration and the antinutrient phytic acid content of polished rice, as a result increased 67.2% ferrtin formation in Caco-2 cell. Moreover, foliar DTPA-Fe application could maintain amylase, protein and minerals quality of polished rice. According to the current study, DTPA-Fe is recommended as an excellent foliar Fe form for Fe biofortification program.

Availability and toxicity of Fe(II) and Fe(III) in Caco-2 cells

The objective of the present study was to compare the toxicity and availability of Fe(II) and Fe(III) to Caco-2 cells. Cellular damage was studied by measuring cell proliferation and lactate dehydrogenase (LDH) release. The activities of two major antioxidative enzymes [superoxide dismutase (SOD) and glutathione peroxidase (GPx)] and differentiation marker (alkaline phosphatase) were determined after the cells were exposed to different levels of iron salts. The cellular iron concentration was investigated to evaluate iron bioavailability. The results show that iron uptake of the cells treated with Fe(II) is significantly higher than that of the cells treated with Fe(III) (P<0.05). Fe(II) at a concentration >1.5 mmol/L was found to be more effective in reducing cellular viability than Fe(III). LDH release investigation suggests that Fe(II) can reduce stability of the cell membrane. The activities of SOD and GPx of the cells treated with Fe(II) were higher than those of the cells treated with Fe(III), although both of them increased with raising iron supply levels. The results indicate that both Fe(II) and Fe(III) could reduce the cellular antioxidase gene expression at high levels.

Folate Content and Composition of Vegetables Commonly Consumed in China

UNLABELLED Folate deficiency increases the risk of chronic diseases, including neural tube defects (NTDs) in infants, megaloblastic anemia, cardiovascular disease, and some cancers in adults. China is the most NTDs prevalent area in the world. Folate deficiency in China can be reduced by proper supply of fresh leafy green vegetables but little is known about the folate content and vitamers in the vegetables commonly consumed by Chinese population. The purposes of this study were first to analyze most commonly consumed important vegetables that contribute to folate intake in the Chinese population and second to estimate the significance of selected vegetables as a source of dietary folate intake. Folate content and vitamers forms in vegetables were analyzed using a valid liquid chromatography method. Monoenzyme treatment was used for leafy green and some fruit vegetables, and dienzyme treatment for some root vegetables. Total folate content in commonly consumed vegetables ranged from 14.78 to 145.54 μg/100 g in edible portion with an average of 61.99 μg/100 g. The highest folate content (>140 μg/100 g) was found in pak choi and spinach. Total folate contents in leafy vegetables, fruit vegetables, and root vegetables were in the range of 17.22 to 145.54 μg/100g, 18.14 to 86.04 μg/100g, and 14.78 to 75.81 μg/100g, respectively. The considerable variations in folate content were found in different types of vegetables commonly consumed by Chinese population. Leafy vegetables are a better source of folate than fruit and root vegetables commonly consumed by Chinese population. PRACTICAL APPLICATION Data from this research would facilitate to accurately establish the actual folate intake by Chinese population. Our folate composition data on vegetables can be incorporated into the national food databases. Availability of such data is essential for estimating folate intake and defining an optimal level for fortification program in China.

A rapid method for sensitive profiling of folates from plant leaf by ultra-performance liquid chromatography coupled to tandem quadrupole mass spectrometer

Previous published methods for the analysis of folates are time consuming because of lengthy sample extraction, clean-up and total running time. This study details the development and validation of a rapid, sensitive and robust method that combines a simple extraction step with ultra-performance liquid chromatography coupled to tandem quadrupole mass spectrometry. Here, we reported application of a tandem quadrupole mass spectrometer to analyze maximum seven vitamers of folate from plant origin. The analytical performance was evaluated by linearity, sensitivity, precision, recovery test and analysis of certified reference materials. The limit of detection and limit of quantification ranged between 0.003 and 0.021μg/100g FW and between 0.011 and 0.041μg/100g FW, respectively; the recovery and precession ranged from 71.27 to 99. 01% and from 1.7 to 7.8% RSD, respectively, depending upon folate vitamers. This newly developed and validated method is rapid (a chromatographic run time of 5min), easy to be performed (no laborious and time consuming clean-up) and can be used to simultaneously analyze seven vitamers of folate from plant sources.