Soil redox status governs within-field spatial variation in microbial arsenic methylation and rice straighthead disease.

Microbial arsenic (As) methylation in paddy soil produces mainly dimethylarsenate (DMA), which can cause physiological straighthead disease in rice. The disease is often highly patchy in the field, but the reasons remain unknown. We investigated within-field spatial variations in straighthead disease severity, As species in rice husks and in soil porewater, microbial composition and abundance of arsM gene encoding arsenite S-adenosylmethionine methyltransferase in two paddy fields. The spatial pattern of disease severity matched those of soil redox potential, arsM gene abundance, porewater DMA concentration, and husk DMA concentration in both fields. Structural equation modelling identified soil redox potential as the key factor affecting arsM gene abundance, consequently impacting porewater DMA and husk DMA concentrations. Core amplicon variants that correlated positively with husk DMA concentration belonged mainly to the phyla of Chloroflexi, Bacillota, Acidobacteriota, Actinobacteriota, and Myxococcota. Meta-omics analyses of soil samples from the disease and non-disease patches identified 5129 arsM gene sequences, with 71% being transcribed. The arsM-carrying hosts were diverse and dominated by anaerobic bacteria. Between 96 and 115 arsM sequences were significantly more expressed in the soil samples from the disease than from the non-disease patch, which were distributed across 18 phyla, especially Acidobacteriota, Bacteroidota, Verrucomicrobiota, Chloroflexota, Pseudomonadota, and Actinomycetota. This study demonstrates that even a small variation in soil redox potential within the anoxic range can cause a large variation in the abundance of As-methylating microorganisms, thus resulting in within-field variation in rice straighthead disease. Raising soil redox potential could be an effective way to prevent straighthead disease.

Reduction of Dimethylarsenate to Highly Toxic Dimethylarsenite in Paddy Soil and Rice Plants.

Dimethylarsenate [DMAs(V)] is a common methylated As species in soils and plants and can cause the physiological disorder straighthead disease in rice. Because DMAs(V) is relatively noncytotoxic, we hypothesize that phytotoxicity of DMAs(V) may arise from trivalent dimethylarsenite [DMAs(III)]. DMAs(III) has been detected in human urine samples but not in environmental samples, likely due to its instability under oxic conditions. We first established methods for preservation and detections of DMAs(III) in soil and plant samples. We showed that DMAs(III) was a major As species in soil solution from an anoxic paddy soil. Enrichment cultures for fermentative, sulfate-reducing, and denitrifying bacteria from the paddy soil could reduce DMAs(V) to DMAs(III). Twenty-two strains of anaerobic bacteria isolated from the soil showed some ability to reduce DMAs(V). Rice plants grown in hydroponic culture with DMAs(V) also showed the ability to reduce DMAs(V) to DMAs(III). Rice plants and grains grown in a flooded paddy soil contained both DMAs(V) and DMAs(III); their concentrations were higher in the spikelets with straighthead disease than those without. DMAs(III) was much more toxic to the protoplasts isolated from rice plants than DMAs(V). Taken together, the ability to reduce DMAs(V) to highly toxic DMAs(III) is common to soil anaerobes and rice plants.

Multi-site field trials demonstrate the effectiveness of silicon fertilizer on suppressing dimethylarsenate accumulation and mitigating straighthead disease in rice.

Rice accumulates both inorganic arsenic (iAs) and organic As species such as dimethylarsenate (DMA). Although DMA is less toxic to humans, it has been shown in hydroponic studies to induce rice straighthead disease, a physiological disorder prevalent in some rice growing regions causing large yield losses. We investigated the effects of different amendments on As species dynamics in soil porewater, accumulation of As species in rice husks and grains, and the incidence of straighthead disease in five field experiments conducted over 2 years at three sites where straighthead disease was observed in previous seasons. The amendments included silicon (Si) fertilizer, micronized zero valent iron (µZVI), sulfate, nitrate, Si-rich biochar, and a mixture of trace and major elements. Straighthead disease was observed in all five experiments. Rice panicles showing the straighthead disease symptoms contained much higher DMA concentrations in husks and grains than normal panicles. Silicon fertilizer was highly effective at decreasing the disease incidence rate and increasing seed setting rate, resulting in 14.9-58.1% increase in grain yield. Silicon fertilizer increased soil porewater iAs and DMA concentrations, but decreased iAs and DMA accumulation in husks and grains, suggesting that Si suppressed the uptake of iAs and DMA by rice plants. Other amendments alleviated straighthead disease to smaller extents than Si fertilizer, with the effect of biochar and the mixture of trace and major elements likely also being attributed to the addition of Si. Results from this field-based study demonstrate that excessive accumulation of DMA is the main cause of rice straighthead disease and Si fertilizer is highly effective at mitigating this disease by suppressing DMA accumulation.

Transformation of arsenic species by diverse endophytic bacteria of rice roots.

Rice growing in flooded paddy soil often accumulates considerable levels of inorganic and organic arsenic (As) species, which may cause toxicity to plants and/or pose a risk to human health. The bioavailability and toxicity of As in soil depends on its chemical species, which undergo multiple transformations driven primarily by soil microbes. However, the role of endophytes inside rice roots in As species transformation remains largely unknown. We quantified the abundances of microbial functional genes involved in As transformation in the endosphere and rhizosphere of rice roots growing in three paddy soils in a pot experiment. We also isolated 46 different bacterial endophytes and tested their abilities to transform various As species. The absolute abundances of the arsenate reductase gene arsC and the dissimilatory arsenate reductase gene arrA in the endosphere were comparable to those in the rhizosphere, whereas the absolute abundances of the arsenite methylation gene arsM and arsenite oxidation gene aioA in the endosphere were lower. After normalization based on the bacterial 16S rRNA gene, all four As transformation genes showed higher relative abundances in the endosphere than in the rhizosphere. Consistent with the functional gene data, all of the 30 aerobic endophytic isolates were able to reduce arsenate, but only 3 strains could oxidize arsenite. Among the 16 anaerobic endophytic isolates, 4 strains belonging to Desulfovibrio, Terrisporobacter or Clostridium could methylate arsenite and/or methylarsenite. Six strains of aerobic endophytes could demethylate methylarsenite, among which three strains also could reduce and demethylate methylarsenate. None of the isolates could demethylate dimethylarsenate. These results suggest that diverse endophytes living inside rice roots could participate in As species transformation and affect As accumulation and species distribution in rice plants.

Widespread Occurrence of the Highly Toxic Dimethylated Monothioarsenate (DMMTA) in Rice Globally.

Arsenic (As) accumulation in rice is of global concern for human health and international trade. Rice is typically reported to contain inorganic As (iAs) and dimethylated arsenate (DMA), with current food guidelines limiting toxic iAs but not less-toxic DMA. Here, we show that the highly toxic dimethylated monothioarsenate (DMMTA) is also found in rice worldwide and has been unknowingly determined as less-toxic DMA by previous routine analytical methods. Using enzymatic extraction followed by high-performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) analysis with a C18 column, DMMTA was detected in rice grains (n = 103) from a field survey from China and in polished rice grains (n = 140) from a global market-basket survey. Concentration ranged from <0.20 to 34.8 µg/kg (median 10.3 µg/kg), accounting for 0 to 21% of total As. A strong linear correlation was observed in all rice samples between DMA and DMMTA (being 30 ± 8% of DMA) concentrations. This robust relationship allows an estimation of DMMTA in rice grains from the DMA data reported in previous market-basket surveys, showing a general global geographical pattern with DMMTA concentration increasing from the equator toward high-latitude regions. Based on the global occurrence and potential high toxicity, DMMTA in rice should be considered in health risk assessments and for setting food regulations.

Dynamics of Dimethylated Monothioarsenate (DMMTA) in Paddy Soils and Its Accumulation in Rice Grains.

Arsenic species transformation in paddy soils has important implications for arsenic accumulation in rice grains and its safety to the consumers. Methylated thioarsenates including highly toxic dimethylated monothioarsenate (DMMTA) have been detected in paddy soils, but their production and dynamics remain poorly understood. In the present study, we first optimized a HPLC-ICP-MS method to quantify methylated thioarsenate species. Using this method together with 10 mM diethylenetriamine pentaacetate (DTPA) to preserve As speciation, we investigated methylated thioarsenate species in porewaters of seven As-contaminated soils incubated under flooded conditions and of two paddy fields. DMMTA was the main methylated thioarsenate species in the porewaters in both incubated soils and paddy fields, with concentrations ranging from 0.2 to 36.2 µg/L and representing ca. 58% of its precursor dimethylarsenate (DMA). The temporal production and dynamics of DMMTA were linked with the DMA concentrations. When soils were drained, DMMTA was converted to DMA. In the two paddy fields, DMMTA concentrations in rice grains were 0.4-10.1 µg/kg. Addition of sulfur fertilizer and rice straw incorporation increased grain DMMTA by 9-28%. These results suggest that DMMTA is an important As species in paddy soils and can accumulate in rice grains, presenting a risk to food safety and human health.

Dimethylarsinic acid is the causal agent inducing rice straighthead disease.

Straighthead disease is a physiological disorder in rice with symptoms of sterile spikelets, distorted husks, and erect panicles. Methylated arsenic species have been implicated as the causal agent of the disease, but direct evidence is lacking. Here, we investigated whether dimethylarsinic acid (DMA) causes straighthead disease and its effect on the transcriptome of young panicles. DMA addition caused typical straighthead symptoms in hydroponic culture, which were alleviated by silicon addition. DMA addition to soil at the tillering to flowering stages induced straighthead disease. Transgenic rice expressing a bacterial arsenite methyltransferase gene gained the ability to methylate arsenic to mainly DMA, with the consequence of inducing straighthead disease. Field surveys showed that seed setting rate decreased with increasing DMA concentration in the husk, with an EC50 of 0.18 mg kg-1. Transcriptomic analysis showed that 364 and 856 genes were significantly up- and down-regulated, respectively, in the young panicles of DMA-treated plants compared with control, whereas Si addition markedly reduced the number of genes affected. Among the differentially expressed genes, genes related to cell wall modification and oxidative stress responses were the most prominent, suggesting that cell wall metabolism is a sensitive target of DMA toxicity and silicon protects against this toxicity.

How Could Agronomic Biofortification of Rice Be an Alternative Strategy with Higher Cost-Effectiveness for Human Iron and Zinc Deficiency in China?

BACKGROUND: Iron and zinc deficiencies affect human health globally, especially in developing countries. Agronomic biofortification, as a strategy for alleviating these issues, has been focused on small-scale field studies, and not widely applied while lacking of cost-effectiveness analysis (CEA). OBJECTIVE: We conducted the CEA of agronomic biofortification, expressed as USD per disability-adjusted life years (DALYs) saved, to recommend a cost-effectiveness strategy that can be widely applied. METHODS: The DALYs were applied to quantify the health burden due to Fe and/or Zn deficiency and health cost of agronomic biofortification via a single, dual, or triple foliar spray of Fe, Zn, and/or pesticide in 4 (northeast, central China, southeast, and southwest) major Chinese rice-based regions. RESULTS: The current health burden by Fe or Zn malnutrition was 0.45 to 1.45 or 0.14 to 0.84 million DALYs for these 4 regions. Compared to traditional rice diets, the daily Fe and/or Zn intake from Fe and/or Zn-biofortified rice increased, and the health burden of Fe and/or Zn deficiency decreased by 28% and 48%, respectively. The cost of saving 1 DALYs ranged from US$376 to US$4989, US$194 to US$2730, and US$37.6 to US$530.1 for the single, dual, and triple foliar Fe, Zn, and/or pesticide application, respectively, due to a substantial decrease in labor costs by the latter 2 applications. CONCLUSIONS: Agronomic biofortification of rice with the triple foliar spray of Fe, Zn, and pesticide is a rapidly effective and cost-effectiveness pathway to alleviate Fe and Zn deficiency for rice-based dietary populations.