Boron deficiency decreased the root activity of Ga-exposed rice seedlings by reducing iron accumulation and increasing Ga in iron plaque.

Gallium (Ga) is an emerging chemical pollutant chiefly associated with high-tech industries. Boron (B) alleviates the negative effects of toxic elements on plant growth. Thereby, the effects of B fertilization on Ga toxicity in rice seedlings was studied to clarify the role of iron plaque in the distribution of Ga, Fe, and B in Ga-treated rice seedlings in the presence or absence of B. Gallium exposure significantly reduced the biomass of rice seedlings. Boron deficiency induced a significant change in the distribution of B in Ga-treated rice seedlings compared with "Ga+B" treatments. Accumulation of Ga in roots, dithionite-citrate-bicarbonate (DCB) extracts, and shoots showed a dose-dependent manner from both +B and -B rice seedlings. Boron nutrition levels affect the distribution of Fe in roots, DCB extracts, and shoots, in which DCB-extractable Fe was significantly decreased from "Ga-B" treatments compared with "Ga+B" treatments. Root activity was significantly decreased in both Ga-exposed rice seedlings; however, B-deficient seedlings showed a severe reduction than +B rice seedlings. These results reveal that Fe plaque might be a temporary sink for B accumulation when plants are grown with proper B, wherein the re-utilization of DCB-extractable B stored in Fe plaque is mandatory for plant growth under B deficiency. Correlation analysis revealed that B deficiency decreased the root activity of Ga-exposed rice seedlings by reducing DCB-extractable Fe and increasing DCB-extractable Ga in Fe plaque. This study enhances our understanding of how B nutritional levels affect Ga toxicity in rice plants.

Proline-mediated modulation on DNA repair pathway in rice seedlings under chromium stress by integrating gene chip and co-expression network analysis.

Chromium (Cr) stress can cause oxidative burst to plants. Application of exogenous proline (Pro) is one of the most effective approaches to improve the tolerance of plants to Cr stress. In this study, we integrated the data of gene chip with co-expression network analysis to identify the key pathways involved in the DNA repair processes in rice seedlings under Cr(VI) stress. Based on KEGG pathway analysis, 158 genes identified are activated in five different types of DNA repair pathways, namely base excision repair (BER, 20 genes), mismatch repair (MMR, 30 genes), nonhomologous end joining (NHEJ, 8 genes), nucleotide excision repair (NER, 56 genes) and homologous recombination (HR, 44 genes). Co-expression network analysis showed that genes activated in DNA repair pathways were categorized into six different modules, wherein Module 1 (45.36%), Module 2 (27.84%) and Module 3 (19.59%) carried more weight than others. Integrating the data of gene chip and co-expression network analysis indicated that coordinated actions of HR and NER pathways are mainly associated with DNA repair processes in Cr(VI)-treated rice seedlings supplied with exogenous Pro. OsCSB, OsXPG, OsBRIP1, OsRAD51C, OsRAD51A2, OsRPA, OsTOPBP1C, OsTOP3, and OsXRCC3 activated in the HR pathway had a stronger impact on repairing DNA damage induced by Cr(VI) stress in rice seedlings supplied with exogenous Pro, while OsXPB1, OsTTDA2, OsTFIIH1, OsXPC, OsRAD23, OsDSS1, and OsRPA located at the NER pathway showed more contribution to repairing DNA damage than others.

Tracing the pollution and human risks of potentially toxic elements in agricultural area nearby the cyanide baths from an active private gold mine in Hainan Province, China.

Mining activities are well-known sources of potentially toxic elements (PTEs) pollution, which often jeopardize the biosphere, pedosphere, and hydrosphere. However, the soil and groundwater pollution caused by active private mining activities has long been neglected. This study investigated the occurrence of PTEs and cyanide (CN) in agricultural soils, mine tailings, and groundwater nearby the cyanide baths from a private gold mine in Hainan Province, southern China. Results indicated that concentrations of Pb, As, Cd, Hg, and CN in different soil depths and mine tailings were up to ten thousand mg/kg, and relatively higher content of As and Pb was detected in groundwater. The chemical forms of Cd, Pb, As, and Hg varied greatly in different soil depths; over 80% of Cd distributed in the water-soluble fraction, suggesting its higher mobility in soils, while approximately 60-90% of Pb, As, and Hg distributed in other chemical fractions, indicating relatively lower mobility in soils. The pollution indices also revealed the serious pollution and deterioration of site quality in this area. Human risk assessments also reflected a high non-carcinogenic/carcinogenic health risk in this area. The framework of integrated management strategies for private metal mines was proposed to mitigate PTEs pollution and reduce health risks.

Estimating the synergistic and antagonistic effects of dual antibiotics on plants through root elongation test.

Antibiotics are recently recognized as a group of emerging environmental contaminants that are frequently detected in various environmental matrixes. Relative root elongation (RRE) test is a rapid and effective strategy to evaluate the water/soil quality and the toxic effects of environmental contaminants on plants. In the present study, we examine the toxicity effect of ciprofloxacin (CIP), norfloxacin (NOR), and tetracycline (TET) to pakchoi individually and in combinations. Both independent action (IA) and concentration addition (CA) models are used for toxicity assessment. Results showed that the EC50 values of CIP, NOR, and TET are 193.59, 60.81, and 40.37 µM, respectively. Combinations of TET + CIP and TET + NOR caused more inhibitory effects on root elongation than those of CIP + NOR. Toxic Unit (TU) and Synergistic Ratio (SR) analysis showed that the relatively lower (higher) EC values are observed in the combinations with lower (higher) antibiotic concentrations, suggesting an effect of low-dose synergism and high-dose antagonism. The reliability of the simulation results from IA and CA models to predict that combined toxicity is highly dependent upon the results from the analysis of TU or SR.

Jasmonic acid and hydrogen sulfide modulate transcriptional and enzymatic changes of plasma membrane NADPH oxidases (NOXs) and decrease oxidative damage in Oryza sativa L. during thiocyanate exposure.

It is evident that the plasma membrane NADPH oxidases (NOXs) play an important role in the generation of superoxide radicals (O2-•) in plants during defense responses. This study was to clarify activation of NOXs in oxidative damage in Oryza sativa during SCN- exposure, particularly in the roles of jasmonic acid (JA) and hydrogen sulfide (H2S) on transcriptional and enzymatic changes of NOXs. Results indicated that enzymatic activity of NOXs in both roots and shoots was significantly enhanced during SCN- exposure, whereas the application of JA and H2S donor (NaHS) significantly repressed NOXs activity in SCN-treated rice seedlings. Similarly, ROS analysis showed that SCN- exposure elevated the content of O2-•, hydrogen peroxide (H2O2) and malondialdehyde (MDA) in rice tissues significantly, whereas decreases in O2-• and H2O2 were detected in roots and shoots of SCN-treated rice seedlings due to application of JA and NaHS. PCR analysis revealed different expression patterns of 7 plasma membrane-localized NOX genes in rice roots and shoots against SCN- exposure, suggesting that various isogenes of NOXs might regulate and determine activity of NOXs in rice organs. In conclusion, SCN- exposure was able to trigger activation of NOXs effectively, and led to oxidative damage and lipid peroxidation; the effects of JA and NaHS on inactivation of NOXs was evident and tissue specific, which in turn modulated ROS accumulation in rice plants.

A modelling study of a buffer zone in abating heavy metal contamination from a gold mine of Hainan Province in nearby agricultural area.

The establishment of new buffer zone in the mine areas lacks the basis of theoretical model. As a case study, this study presents the partition model of new buffer zone in a small-scale mine area located in the remote mountainous region of Hainan province, China. The investigation carried out by integration of geostatistical interpolation maps using unmanned aerial vehicle has revealed that As, Cd, Pb, Zn, and Hg are mainly distributed near the tailing ponds and the dressing plant, while Cr and Cu are prominent near the agricultural area. The partition model of buffer zone was established in accordance to the transport velocity of the liquid phase heavy metal in soil. The boundary line of buffer zone is integrated by comparing the 'predicted' heavy metal concentrations with the 'measured' heavy metal concentrations. Principal component analysis (PCA) and the variations of heavy metal concentrations with elevation, slope, and distance were performed to further evaluate the reliability of the predicted buffer zone, indicating that the new buffer zone has a significant effect on minimizing transport of heavy metals from the mining area into the agricultural land. This study is suggestive to provide risk control and sustainable development strategy for the ecosystem protection in mine areas.

Transcriptomic analysis of cytochrome P450 genes and pathways involved in chromium toxicity in Oryza sativa.

In plants, cytochrome P450 monooxygenase (CYP) plays an important role in detoxifying xenobiotic chemicals and coordinating abiotic stresses. Agilent 44 K rice microarray has been used to focus on the transcriptional profile of osCYP genes in rice seedling exposed to Cr solution containing K2CrO4 or Cr(NO3)3. Our study showed that expression profiles of 264 osCYP genes identified were tissue, dose and stimulus specific in rice seedlings. Comparative genomics analysis revealed that more differentially expressed osCYP genes were discovered in roots than in shoots under both Cr exposures. Results from Venn diagram analysis of differentially expressed osCYP genes demonstrated that there were common osCYP genes and unique osCYP genes present in different rice tissue as well as in different Cr treatments, which may control and/or regulate involvement of different CYP isoenzymes under Cr exposure individually or combinedly. KEGG analysis indicated that significant up- and down-regulated osCYP genes in rice tissues were chiefly related to "biosynthesis of secondary metabolites". However, involvements of osCYP genes mapped in the "biosynthesis of secondary metabolites" were tissue and dose specific, implying their distinctly responsive and adaptive mechanisms during Cr exposure. Overall, our findings are evident to describe and clarify their individual roles of specific osCYP genes in regulating involvement of CYP isoforms in Cr detoxification by rice seedlings.

Assimilation of exogenous cyanide cross talk in Oryza sativa L. to the key nodes in nitrogen metabolism.

Exogenous cyanide (CN-) effects on nitrogen (N) uptake, transport, and assimilation in rice seedlings were investigated at the biochemical and molecular levels. Seedlings were treated with either a 2-d or 4-d supply of potassium cyanide (KCN) in the nutrient solution containing nitrate (NO3-) or ammonium (NH4+). Although a KCN-induced increase was recorded in the activity of ß-cyanoalanine synthase (ß-CAS) in rice tissues of both NH4+-fed and NO3--fed seedlings, the former showed a significantly greater assimilation rate for CN- than the latter. The addition of KCN decreased NO3- uptake and assimilation, whereas a negligible impact was observed in NH4+ treatments. Enzymatic assays showed a marked activities enhancement of glutamine synthetase (GS), glutamate synthase (GOGAT), and glutamate dehydrogenase (GDH) in NH4+-fed seedlings after KCN exposure. Similarly, the rises occurring in nitrate reductase (NR) and GDH activity in NO3--fed seedlings were also detected after 2-d exposure to KCN, whereas a significant reduction of GS and GOGAT activities was determined. The results suggest that although exogenous KCN at moderate or high concentrations caused repressively effects on biomass growth of both NH4+-fed and NO3--fed rice seedlings, the nontoxic concentration of KCN supplied can serve as a supplemental N source in plant nutrition and N metabolism.

Involvement of glutamate receptors in regulating calcium influx in rice seedlings under Cr exposure.

Glutamate receptors (GLRs) are ligand-gated Ca2+-permeable channels that govern and modulate the dynamic influx of cytosolic Ca2+ in plants. The present study investigated the interaction of OsGLR3 gene expression with subcellular Ca distribution in rice seedlings exposed to chromium (Cr) solution containing Cr(III) or Cr(VI). The results displayed that the accumulation of Ca was evaluated or higher in shoots compared to roots under Cr exposure, and a similar pattern of subcellular Ca distribution was observed between rice tissues exposed to Cr(III) and Cr(VI). Real-time quantitative polymerase chain reaction (qRT-PCR) analysis revealed that eight OsGLR3 isogenes were distinctly expressed in different rice tissues at different levels of Cr exposures. This differential expressions could possible be due to the uptake variations, subcellular distribution and chemical speciation of the two Cr species. Notably, distinct expression patterns of OsGLR3 genes were found between Cr(III) and Cr(VI) exposures, suggesting that different regulation strategies are used to mediate Ca influx in rice materials under different Cr exposures. These results demonstrated a full picture of Cr-induced transcriptional alterations in OsGLR3 expression levels in rice seedlings, and provided suggestive evidence for further investigation on specific OsGLR3 genes participated in the regulation of cytosolic Ca2+ concentrations under Cr exposure.

Differential expression of the PAL gene family in rice seedlings exposed to chromium by microarray analysis.

Phenylalanine ammonia-lyase (PAL) is one of the principle enzymes involved in plant's secondary metabolism. Expression of individual isogene from the PAL gene family is variable with species of plants in responses to different stresses. In this study, transcriptome analysis of the PAL gene family in rice seedlings exposed to potassium chromate Cr(VI) or chromium nitrate Cr(III) was conducted using Agilent 44K rice microarray and real-time quantitative RT-PCR. Uptake and accumulation of both Cr species by rice seedlings and their effect on PAL activity were also determined. Three days of Cr exposure led to significant accumulation of Cr in plant tissues, but majority being in roots rather than shoots. Changes of PAL activities in rice tissues were evident from both Cr treatments. Individual isogene from the rice PAL gene family was expressed differentially in response to both Cr variants. Comparing gene expression between two Cr treatments, only osPAL2 and osPAL4 genes were expressed in similar patterns. Also, gene expression pattern was inconsistent in both plant tissues. Results indicated that expression of individual isoform from the rice PAL gene family is tissue, and stimulus specific under different Cr exposure, suggesting their different detoxification strategies for decreasing or eliminating Cr stresses.

mRNA Analysis of Genes Encoded with Phytochelatin Synthase (PCS) in Rice Seedlings Exposed to Chromium: The Role of Phytochelatins in Cr Detoxification.

Transcriptional changes of genes encoded with phytochelatins synthase (PCS) was investigated in rice seedlings exposed to potassium chromate Cr(VI) or chromium nitrate Cr(III) using qRT-PCR. Our study demonstrates that both Cr variants initiated different responses of phytochelatin content and PCS activities in rice tissues. Six putative PCS genes were expressed differentially in response to both Cr species. Comparing gene expression between root/shoots, only LOC_Os05g34290.1 and LOC_Os06g01260.1 genes were expressed in similar patterns in Cr(VI) treatment, while none of them were expressed equally in Cr(III) treatments. Inconsistent expression of PCS genes in two Cr variants as well as in rice tissues were most likely related to its individual chemical properties and chemical speciation. Results presented here indicate that the role of phytochelatins in Cr detoxification between two Cr variants in rice was different and six putative PCS genes functioned differently in stimulating PCS activities and regulating phytochelatin formation.

Identification and expression analysis of CYS-A1, CYS-C1, NIT4 genes in rice seedlings exposed to cyanide.

Involvement of genes (CYS-A1, CYS-C1 and NIT4) encoded with cysteine synthase, ß-cyanoalanine synthase, nitrilase and cyanide metabolisms are evident in Arabidopsis. In the present study, identifications of CYS-A1, CYS-C1 and NIT4, predictions of conserved motifs, and constructions of phylogenetic relationships, based on their amino acid sequences in rice, were conducted. In order to elucidate the transcriptional responses of these cyanide-degrading genes, two candidate homologues were selected for each gene to test their expression changes upon exposure to exogenous KCN in rice seedlings using RT-PCR. Results showed that all selected candidate homologous genes were differentially expressed at different exposure points in roots and shoots of rice seedlings, suggesting their distinct roles during cyanide assimilation. Both candidate homologues for CYS-A1 constantly exhibited more abundant transcripts in comparison to control. However, only one candidate homologue for CYS-C1 and NIT4 showed a remarkable up-regulation during KCN exposure. Analysis of both tissue and solution cyanide indicated that rice seedlings were quickly able to metabolize exogenous KCN with minor accumulation in plant tissues. In conclusion, significant up-regulation of CYS-A1 suggested that the endogenous pool of cysteine catalyzed by cysteine synthase does not restrict the conversion of exogenous KCN into cyanoalanine through the ß-cyanoalanine pathway. However, insufficient responses of the transcription level of NIT4 suggested that NIT enzyme may be a limiting factor for cyanoalanine assimilation by rice seedlings.

Determination of the Michaelis-Menten kinetics and the genes expression involved in phyto-degradation of cyanide and ferri-cyanide.

Hydroponic experiments were conducted with different species of plants (rice, maize, soybean and willow) exposed to ferri-cyanide to investigate the half-saturation constant (K M ) and the maximal metabolic capacity (v max ) involved in phyto-assimilation. Three varieties for each testing species were collected from different origins. Measured concentrations show that the uptake rates responded biphasically to ferri-cyanide treatments by showing increases linearly at low and almost constant at high concentrations from all treatments, indicating that phyto-assimilation of ferri-cyanide followed the Michaelis-Menten kinetics. Using non-linear regression, the highest v max was by rice, followed by willows. The lowest v max was found for soybean. All plants, except maize (DY26) and rice (XJ12), had a similar K M value, suggesting the same enzyme was active in phyto-assimilation of ferri-cyanide. Transcript level, by real-time quantitative PCR, of enzymes involved in degradation of cyanides showed that the analyzed genes were differently expressed during different cyanides exposure. The expression of CAS and ST genes responded positively to KCN exposure, suggesting that ß-CAS and ST pathways were two possible pathways for cyanide detoxification in rice. The transcript level of NIT and ASPNASE genes also showed a remarkable up-regulation to KCN, implying the contribution to the pool of amino acid aspartate, which is an end product of CN metabolism. Up-regulation of GS genes suggests that acquisition of ammonium released from cyanide degradation may be an additional nitrogen source for plant nutrition. Results also revealed that the expressions of these genes, except for GS, were relatively constant during iron cyanide exposure, suggesting that they are likely metabolized by plants through a non-defined pathway rather than the ß-CAS pathway.

Phytotoxicity and Transport of Gallium (Ga) in Rice Seedlings for 2-Day of Exposure.

Hydroponic experiments were conducted with rice seedlings to investigate the accumulation and phytotoxicity of gallium nitrate. A linear decrease in relative growth rate, transpiration rate and water use efficiency was observed in rice seedlings with increasing Ga concentrations. However, inhibition of these selected parameters was noted different at different Ga treatments. Relative growth rate was more sensitive towards Ga treatments. Phyto-transport of Ga was apparent, but recovery of Ga in different parts of rice seedlings varied significantly: roots were dominant site for Ga accumulation. The total accumulation rates of Ga were positively correlated to Ga concentrations. Results indicated that the addition of Ga did not cause deleterious effects on plant physiological functions over a 2-day exposure period. Large amounts of Ga were removed from the hydroponic solution through rice seedlings. Accumulation of Ga in plant tissues resulted in growth inhibition of rice seedlings.

Kinetics for adsorptive removal of chromium(VI) from aqueous solutions by ferri hydroxide/oxohydroxides.

Adsorption behaviors of chromate(VI) on two different iron oxides were investigated through batch experiments under different operational conditions. The operational variables studied included sorbent doses, sorbent species, solution pH, contact time, and initial metal ion concentrations. Both ferri hydroxide and ferri oxohydroxides were capable of removing Cr(VI) from aqueous solution effectively. The extent of metal ion removed from aqueous solution by both sorbents increased with increasing contact time and initial metal ion concentrations, while decreased with elevating solution pH initially. Sorption equilibrium time was observed in 150 min. Equilibrium isotherm data were analyzed using the Langmuir and Freundlich isotherms. The Freundlich model yields a better fit than the Langmuir model. Although kinetic measurements with both sorbents had a similar pattern, ferri hydroxide always showed higher sorptive potential for Cr(VI) than ferri oxohydroxides. Results also indicated that sorptive removal of Cr(VI) by both sorbents was observed to be negative in response to solution pH, due to changes in speciation of Cr(VI) in aqueous solution. At pH lower than 3.0, both iron oxides exhibited much greater capacities of removing Cr(VI) from aqueous solution than other treatments. Therefore, it is to conclude that adsorptive removal of Cr(VI) by both iron oxides is effective. Adsorption capacity is mainly determined by speciation of Cr(VI) in solutions, which is controlled and driven by solution pH.

Chelator-induced phytoextraction of zinc and copper by rice seedlings.

Solution culture was carried to investigate capacity of synthetic aminopolycarboxylic acids (ethylenediamine tetraacetate, N-hydroxyethylenediaminetriacetic acid, and diethylenetriamine-pentaacetate) for enhancing botanical removal and transport of heavy metals (Cu and Zn) by plants. Biodegradable organic acids (citric acid, malic acid, and oxalic acid) were also selected as alternatives to compare them with synthesized chelating agents for effectiveness. Young rice seedlings (Oryza sativa L. cv. XZX 45) were grown in nutrient solutions treated with single or combined metal solutions in presence or absence of chelating compounds. Calculation by chemical equilibrium program VISUAL MINTEQ showed that different chelating compounds had various complex potential with Cu(2+) and Zn(2+) ions, in which synthetic chelators exhibited higher complexed capability than biodegradable organic acids. All applied synthetic aminopolycarboxylic acids significantly decreased removal of metal from nutrient solution (p < 0.01), while more or less effects of organic acids supplied on biosorptive potential were observed with all treatments (p > 0.05), compared with the treatment without metal ligands. Synthetic aminopolycarboxylic acids significantly decreased metal concentrations in plant materials in all treatments (p < 0.01). However, biodegradable organic acids decreased metal concentrations in roots (p < 0.01), but enhanced them in shoots (p < 0.01). Results obtained indicated that synthetic aminopolycarboxylic acids decreased uptake of metals by rice seedlings, but translocation of metals complexed within plant materials was evident. Although exogenous biodegradable organic acids showed negligible effect on botanical removal of metals, metals complexed with organic acids was more mobile than those complexed with other chelating agents. These information collected here had important implication for the use of biodegradable metal chelators in transport of essential micronutrients in plant nutrition.

Boron deficiency energizes cyanide uptake and assimilation through activating plasma membrane H+-ATPase in rice plants.

The effect of boron (B) deficiency on mediating the contribution of H+-ATPase in the uptake and assimilation of exogenous cyanide (CN-) is investigated. Under CN- treatments, rice seedlings with B-deficient (-B) conditions exhibited significantly higher CN- uptake and assimilation rates than B-supplemented (+B) seedlings, whereas NH4+ uptake and assimilation rates were slightly higher in -B rice seedlings than in +B. In this connection, the expression pattern of genes encoding ß-CAS, ST, and H+-ATPase was assessed to unravel their role in the current scenario. The abundances of three ß-CAS isogenes (OsCYS-D1, OsCYS-D2, and OsCYS-C1) in rice tissues are upregulated from both "CN--B" and "CN-+B" treatments, however, only OsCYS-C1 in roots from the "CN--B" treatments was significantly upregulated than "CN-+B" treatments. Expression patterns of ST-related genes (OsStr9, OsStr22, and OsStr23) are tissue specific, in which significantly higher upregulation of ST-related genes was observed in shoots from "CN--B" treatments than "CN-+B" treatments. Expression pattern of 7 selected H+-ATPase isogenes, OsA1, OSA2, OsA3, OsA4, OsA7, OsA8, and OsA9 are quite tissue specific between "CN-+B" and "CN--B" treatments. Among these, OsA4 and OsA7 genes were highly activated in the uptake and assimilation of exogenous CN- in -B nutrient solution. These results indicated that B deficiency disturbs the pattern of N cycles in CN--treated rice seedlings, where activation of ST during CN- assimilation decreases the flux of the innate pool of NH4+ produced from CN- assimilation by the ß-CAS pathway in plants. Collectively, the B deficiency increased the uptake and assimilation of exogenous CN- through activating H+-ATPase.

Effects of exogenous thiocyanate on mineral nutrients, antioxidative responses and free amino acids in rice seedlings.

The effects of exogenous thiocyanate (SCN(-)) on amino acids composition, content of mineral nutrients and antioxidative systems in plants were investigated. Young rice seedlings (Oryza sativa L. cv. XZX 45) were grown in nutrient solutions amended with potassium thiocyanate (KSCN). Activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) in plant materials were analyzed in vivo. Mineral nutrients and free amino acids in rice seedlings were also measured to determine metabolic responses to SCN(-) exposure. A significant reduction in transpiration and relative growth was recorded with all treatments (p < 0.05), while changes of total chlorophyll content in leaves was negligible (p > 0.05). SCN-induced toxicity appeared to be more sensitive to activities of POD in shoots and APX activities in roots than the others. The content of nutrient elements in rice seedlings exposed to exogenous SCN(-) was variable, while the effects were more evident at the highest SCN-treatment (p < 0.05). Although the change of total free amino acids in shoots of SCN-exposed seedlings was negligible (p > 0.05), responses of different amino acids to SCN(-) application were quite different. Among fifteen free amino acids detected, serine (Ser), proline (Pro), and methionine (Met) increased, while asparagine (Asp) decreased with an increase of the doses of SCN(-) supplied. Phyto-transport of SCN(-) was apparent and the removal rates were positively correlated to the doses, suggesting that phyto-assimilation of SCN(-) is an enzymatic process through a potentially un-identified degradation pathway.

Exogenous thiocyanate inhibits sulfurtransferase pathway and induces ß-cyanoalanine synthase pathway to enhance exogenous cyanide assimilation in rice plants.

Cyanide (CN-) assimilation in plants takes place by ß-cyanoalanine synthase (ß-CAS) and sulfurtransferase (ST), in which the ST pathway converts CN- into thiocyanate (SCN-). Both chemicals (CN- and SCN-) are frequently detected in the effluent of gold mining operations. In this connection, exogenous SCN- was applied to rice plants with CN- and compared with CN- alone to investigate its effects on CN- assimilation and degradation pathways. Interestingly, the CN- and SCN- content in both roots and shoots were increased with the increase in "CN-" treatments, but surprisingly their content under "SCN-+CN-" treatments did not show the similar trend. The increasing trend remained the same for CN- but the SCN- content was constant with increasing CN- concentrations in comparison with the control (SCN- alone). Additionally, the assimilation rates of CN- in rice plants under "SCN-+CN-" treatments were significantly higher than "CN-" treatments. The application of SCN- with CN- mostly alters the expression of both ß-CAS and ST-associated genes. On one side, the application of SCN- significantly repressed the expression of genes encoded with ST in rice plants, but on the other side, it significantly up-regulated the expression of the ß-CAS gene located in mitochondria. These results reveal that the application of exogenous SCN- increases CN- assimilation rates by inhibiting the ST pathway and stimulating the ß-CAS pathway. This study would provide new insight into the positive effects of exogenous SCN- in increasing CN- assimilation by altering the degradation pathways in rice plants.

Evaluating the significance of amino acids (AAs) in cyanide-treated rice plants under different nitrogen fertilization using the relative importance index of AA.

The biosynthesis of amino acids (AAs) in plants is affected by different nitrogen (N) sources. The effects of exogenous cyanide (KCN) on the concentrations and profiles of AAs in rice seedlings were carried out in the presence of nitrate (+NO3-)/ammonium (+NH4+) or N deficiency (-N). Targeted metabolomics analysis indicated that the highest accumulation of AAs in CN--treated rice seedlings was detected in the "CN-+NH4+" treatments than in other treatments, wherein the doses of exogenous KCN did not significantly affect the total amount of AAs in rice seedlings at the same N fertilized condition. The total content of AAs in rice shoots under "CN-+NH4+" treatments was higher than other treatments, while the total content of AAs in rice roots under "CN-+NO3-" treatments was higher than other treatments. Also, the profiles of 21 AAs in CN--treated rice seedlings showed tissue-specific under different N fertilization. The relative importance index (RII) of AA was used to evaluate the importance of AAs in CN--treated rice seedlings under different N fertilization. The common AAs with higher RII values were compared between three different treatments of KCN (e.g., 0, 1, and 2 mg CN/L). Under "CN-+(-N)" treatments, Ala, Asp, Glu, Val, and Gly (Ala, Gly, Val, and Lys) were the common AAs in rice roots (shoots). Under "CN-+NO3-" treatments, Ala, Glu, Asp, Ser, and Thr (Asp, Ala, Thr, Ser, and Asn) were the common AAs with higher RII values in rice roots (shoots) between all CN- treatments. Under "CN-+NH4+" treatments, Asp, Gln, Asn, and Ala (Asp, Glu, and Thr) were the common AAs with higher RII values in rice roots (shoots) between all CN- treatments. These results suggested that using the RII to describe the change and fluctuation of AAs in rice plants may reflect the different N utilization strategies in response to exogenous CN- exposure.