Generation of low-cadmium rice germplasms via knockout of OsLCD using CRISPR/Cas9

The OsLCD gene, which has been implicated in cadmium (Cd) accumulation in rice, might be a useful target for CRISPR/Cas9 editing. However, the effects of OsLCD gene editing on Cd accumulation, plant growth, and yield traits remain unknown. Here, we used CRISPR/Cas9 to generate oslcd single mutants from indica and japonica rice cultivars. We also generated osnramp5 single mutants and oslcd osnramp5 double mutants in the indica background. When grown in Cd-contaminated paddy soils, all oslcd single mutants accumulated less Cd than the wild types (WTs). Consistent with this, oslcd single mutants grown in Cd-contaminated hydroponic culture accumulated significantly less Cd in the shoots as compared to WTs. This decrease in accumulation probably resulted from the reduction of Cd translocation under Cd stress. Oxidative damage also decreased, and plant growth increased in all oslcd single mutant seedlings as compared to WTs in the presence of Cd. Plant growth and most yield traits, as well essential element concentrations in rice seedling shoots, brown rice, and rice straw, were similar between oslcd single mutants and WTs. In the presence of Cd, Cd concentrations in the brown rice and shoots of oslcd osnramp5 double mutants were significantly decreased compared with WTs as well as osnramp single mutants. Our results suggested that OsLCD knockout may reduce Cd accumulation alone or in combination with other knockout mutations in a variety of rice genotypes; unlike OsNramp5 mutations, OsLCD knockout did not reduce essential element contents. Therefore, OsLCD knockout might be used to generate low-Cd rice germplasms.     

The risks of sulfur addition on cadmium accumulation in paddy rice under different water-management conditions

Recently, the application of sulfur (S) has been recommended to control the accumulation of cadmium (Cd) in rice in contaminated paddy soil. However, the effects of exogenous S on Cd transfer in paddy rice systems under different water-management practices have not been systematically investigated. Pot experiments were performed to monitor the composition of soil pore water and the Cd accumulation in iron plaque and rice tissue were compared under different S (0 and 200 mg/kg Na₂SO₄) and water (continuous and discontinuous flooding) treatments. Sulfur application significantly increased Cd concentrations in soil pore water under discontinuous flooding conditions, but slightly reduced them under continuous flooding. Moreover, the oxidation/reduction potential (Eh) was the most critical factor that affected the Cd levels. When the Eh exceeded −42.5 mV, S became the second critical factor, and excessive S application promoted Cd dissolution. In addition, S addition elevated the Cd levels in iron plaque and reduced the Cd transfer from the iron plaque to rice roots. In rice, S addition inhibited Cd transfer from the rice roots to the straw; thus, more Cd was stored in the rice roots. Nevertheless, additional S application increased the Cd content in the rice grains by 72% under discontinuous flooding, although this effect was mitigated by continued flooding. Under simulated practical water management conditions, S addition increased the risk of Cd contamination in rice, suggesting that S application should be reconsidered as a paddy fertilization strategy.     

Watering techniques and zero-valent iron biochar pH effects on As and Cd concentrations in rice rhizosphere soils, tissues and yield

Zero-valent iron amended biochar (ZVIB) has been proposed as a promising material in immobilizing heavy metals in paddy fields. In this study, the impacts of pH of ZVIB (pH 6.3 and pH 9.7) and watering management techniques (watering amount in the order of CON (control, 5/72)>3/72>1–3/72>3/100>1/72, with 5/72, for example, representing irrigation given to 5 cm above soil surface in 72 hr regular interval) on As and Cd bioavailability for rice and its grain yield (Yield_BR) were investigated in a pot experiment. Brown rice As (As_BR) content was irrelative to the watering treatments, while significantly decreased (>50%) with the addition of both ZVIB materials. The diminutions of brown rice Cd (Cd_BR) content as well as the Yield_BR were highly dependent on both the soil amendment materials’ pH and watering amount. Among all the watering treatments, 3/72 treatment (15% less irrigation water than the CON) with ZVIB 6.3 amendment was the optimum fit for simultaneous reduction of As_BR (50%) and Cd_BR contents (19%) as well as for significant increment (12%) of the Yield_BR. Although high pH (9.7) ZVIB application could also efficiently decrease As and Cd contents in brown rice, it might risk grain yield lost if appropriate (e.g. 3/72 in our study) watering management technique was not chosen. Therefore, ZVIB would be an environmentally friendly option as an amendment material with proper selection of watering management technique to utilize As and Cd co-contaminated arable soils safely for paddy cultivation.     

Roles of soluble minerals in Cd sorption onto rice straw biochar

Transforming to biochar provides an environmentally friendly approach for crop residue reutilization, which are usually applied as sorbent for heavy metal removal. As typical siliconrich material, the specific sorptive mechanisms of rice straw derived biochar(RSBC) are concerned, especially at the low concentration range which is more environmentally relevant. In the present study, Cd sorption onto RSBCs at the concentration of ≤ 5 mg/L was investigated. The sorptive capacity was positively corr...     

Effects of co-exposure to lead and manganese on learning and memory deficits

Lead (Pb) and manganese (Mn) are common neurotoxins. However, individuals are subject to co-exposures in real life, and it is therefore important to study these metals in combination. Weaning Sprague-Dawley rats were given ad libitum access to drinking water solutions containing Pb (100 mg/L), Mn (2.5 mg/mL) or a mixture, and each treatment had its own minocycline (50 mg/(kg•day)) supplement group. The results showed a significant difference in spatial memory and induction levels of hippocampal long-term potentiation (LTP) in all exposure groups when compared with controls. The combined-exposure group exhibited the most pronounced effect when compared with each of the single-metal exposure groups. Microglia displayed activation at day 3 after exposure alone or in combination, while astrocytes showed activation at day 5, accompanied by decreased expression levels of GLAST, GLT-1, and GS. Furthermore, the levels of glutamate in the synaptic cleft increased significantly. When microglial activation was inhibited by minocycline, the activation of astrocytes and the expression of GLAST, GLT-1, and GS were both reversed. In addition, upon minocycline treatment, hippocampal LTP impairment and cognitive injury were significantly alleviated in each of the exposure groups. These results suggest that combined exposure to Pb and Mn can cause greater effects on cognition and synaptic plasticity when compared to single-metal exposure groups. The reason may involve abnormal activation of microglia leading to excessive regulation of astrocytes, resulting in glutamate reuptake dysfunction in astrocytes and leading to perturbed cognition and synaptic plasticity.     

Accumulation, transportation, and distribution of tetracycline and cadmium in rice

Co-exposure to heavy metal and antibiotic pollution might result in complexation and synergistic interactions, affecting rice growth and further exacerbating pollutant enrichment. Therefore, our study sought to clarify the influence of different Tetracycline (TC) and Cadmium(Cd) concentration ratios (both alone and combined) on rice growth, pollutant accumulation, and transportation during the tillering stage in hydroponic system. Surprisingly, our findings indicated that the interaction between TC and Cd could alleviate the toxic effects of TC/Cd on aerial rice structures and decrease pollutant burdens during root elongation. In contrast, TC and Cd synergistically promoted the accumulation of TC/Cd in rice roots. However, their interaction increased the accumulation of TC in roots while decreasing the accumulation of Cd when the toxicant doses increased. The strong affinity of rice to Cd promoted its upward transport from the roots, whereas the toxic effects of TC reduced TC transport. Therefore, the combined toxicity of the two pollutants inhibited their upward transport. Additionally, a low concentration of TC promoted the accumulation of Cd in rice mainly in the root tip. Furthermore, a certain dose of TC promoted the upward migration of Cd from the root tip. Laser ablation-inductively coupled plasma mass spectrometry demonstrated that Cd mainly accumulated in the epidermis and stele of the root, whereas Fe mainly accumulated in the epidermis, which inhibited the absorption and accumulation of Cd by the rice roots through the generation of a Fe plaque. Our findings thus provide insights into the effects of TC and Cd co-exposure on rice growth.     

Cultivar-dependent rhizobacteria community and cadmium accumulation in rice: Effects on cadmium availability in soils and iron-plaque formation

The association between the rhizospheric microbial community and Cd accumulation in rice is poorly understood. A field trial was conducted to investigate the different rhizobacterial communities of two rice cultivars with high Cd accumulation (HA) and low Cd accumulation (LA) at four growth stages. Results showed that the Cd content in the roots of the HA cultivar was 1.23 - 27.53 higher than that of the LA cultivar (0.08 - 10.5 µg/plant) at four stages. The LA cultivar had a significantly lower Cd availability in rhizosphere and a higher quantity of iron plaque (IP) on the root surface than the HA cultivar at four stages. This resulted in the reduction of Cd concentration in IPs and Cd translocation from IP-to-root. Microbial analysis indicated that the LA cultivar formed a distinct rhizobacterial community from the HA cultivar and had less α-diversity. The rhizosphere of the LA cultivar was enriched in specific bacterial taxa (e.g., Massilia and Bacillus) involved in Cd immobilization by phosphate precipitation and IP formation by iron oxidization. However, the rhizosphere in the HA cultivar assembled abundant sulfur-oxidizing bacteria (e.g., Sulfuricurvum) and iron reduction bacteria (Geobacter). They promoted Cd mobilization and reduced IP formation via the metal redox process. This study reveals a potential approach in which specific rhizobacteria decrease or increase Cd accumulation in rice on contaminated soil and provides a new perspective for secure rice production.     

Hygroscopic properties of sodium and potassium salts as related to saline mineral dusts and sea salt aerosols

Mineral dust,soil,and sea salt aerosols are among the most abundant primary inorganic aerosols in the atmosphere,and their hygroscopicity affects the hydrological cycle and global climate.We investigated the hygroscopic behaviors of six Na-and K-containing salts commonly found in those primary organic aerosols.Their hygroscopic growths as a function of relative humidity(RH) agree well with thermodynamic model prediction.Temperature dependence of deliquescence RH(DRH) values for five of those sal...