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.

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.

Response to gallium (Ga) exposure and its distribution in rice plants.

Root architecture is the temporal and spatial configuration of root system in the heterogeneous matrix of soil that is prone to chemical stresses. Gallium (Ga) is among the emerging chemical pollutants that are mostly associated with high-tech industries, specifically associated with semiconductors. In view of its potential risk and increasing distribution in the environment, this study was designed to evaluate the inhibition rate, Ga distribution in different tissues, and root architecture of rice seedlings under different concentrations of Ga. We observed that 2.59, 46.7, and 168.2 mg Ga/L were minimum (EC20), medium (EC50), and maximum (EC75) effective concentrations for rice plants that corresponded to the 20, 50, and 75% inhibition on the relative growth rate, respectively. Distribution of Ga in rice tissues showed that accumulation of Ga was much higher in roots than shoots of rice seedlings, and it increased with an increase in Ga doses. Evan blue staining technique reveals that the number of damaged/dead cell was dose-dependent on Ga. Moreover, several traits associated with root system architecture demonstrating that rice root system architecture altered in response to Ga stress. Collectively, the results reveal that Ga exposure inhibited the growth and development of rice plants. This study will enhance our understanding that how different concentrations of Ga in the environment can affect plants; however, more comprehensive studies are essential to further determine plant response against Ga stress.

Mathematical Estimation of Endogenous Proline as a Bioindicator to Regulate the Stress of Trivalent Chromium on Rice Plants Grown in Different Nitrogenous Conditions.

The accumulation of proline impacts the defense mechanisms of plants against the harmful effects of adverse environmental conditions; however, its concentration in plants is associated with the metabolism of N. Therefore, the effects of exogenous organic [glutamate (Glu)/arginine (Arg)] and inorganic [nitrate (NO3-)/ammonium (NH4+)] N on the accumulation of proline (Pro) in rice plants under trivalent chromium [Cr(III)] stress were studied through using the mass balance matrix model (MBMM). Application of 'NH4+' showed the largest contribution to the Pro content in rice shoots under different concentrations of Cr(III), followed by 'NO3-', 'Arg', and 'Glu' applications. On the other hand, 'Arg' application displayed the largest contribution to the Pro content in roots under Cr(III) stress, followed by 'NH4+', 'Glu', and 'NO3-' applications. The combined application of 'NH4++Arg' showed the greatest contribution to the Pro content in both roots and shoots of Cr(III)-treated rice seedlings, while the application of 'NO3-+Glu' showed the least contribution to the Pro content in rice seedlings. The current study indicated that the endogenous level of Pro in rice seedlings is quite sensitive to Cr(III) stress under different N sources, and the mathematical modeling showed a reliable result while estimating the relationship between Pro content and N source application.

Proline interacts with Ca2+-dependent signaling to enhance chromium tolerance in rice by manipulating nitrate reductase and sucrose phosphate synthase.

The entrance of chromium (Cr) into the agricultural system would exert a negative influence on the carbon/nitrogen metabolism (CNM) of plants. In this study, we investigated the role of exogenous proline-mediated Ca2+-dependent signaling in the regulation of CNM in rice subjected to Cr(VI) stress, with emphasis on the involvement of nitrate reductase (NR) and sucrose phosphate synthase (SPS). Results demonstrated that proline effectively mitigated the growth inhibition of rice imposed by Cr(VI) stress, which is achieved by a reduction in cytoplasmic Ca and Cr content and the activation of the downstream Ca2+-dependent signaling pathway. Additionally, proline displayed a positive effect in modulating the expression and activities of NR and SPS under Cr(VI) stress, which are attributed to the cross-regulation between calcium-dependent protein kinases (CDPKs) and 14-3-3 proteins (14-3-3s). Consequently, nitrogen use efficiency and sucrose content in rice under Cr(VI) + proline treatments were higher than Cr(VI) treatments. Gene expression variation factors underscored that the regulation of proline on NR is crucial to the Ca2+-dependent signaling pathway, initiated by the interaction between CDPKs and 14-3-3s in rice plants during Cr(VI) stress. These results reveal that proline interacts with Ca2+-dependent signaling pathways to enhance Cr tolerance in rice by regulating NR and SPS.

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.

Proline-mediated activation of glyoxalase II improve methylglyoxal detoxification in Oryza sativa L. under chromium injury: Clarification via vector analysis of enzymatic activities and gene expression.

Environmental factors affect plants in several ways including the excessive accumulation of methylglyoxal (MG), resulting in dysfunctions of many biological processes. Exogenous proline (Pro) application is one of the successful strategies to increase plant tolerance against various environmental stresses, including chromium (Cr). This study highlights the alleviation role of exogenous Pro on MG detoxification in rice plants induced by Cr(Vl) through modifying the expression of glyoxalase I (Gly I)- and glyoxalase II (Gly II)-related genes. The MG content in rice roots was significantly reduced by Pro application under Cr(VI) stress, however, there was little effect on the MG content in shoots. In this connection, the vector analysis was used to compare the involvement of Gly l and Gly II on MG detoxification in 'Cr(VI)' and 'Pro+Cr(VI)' treatments. Results exhibited that vector strength in rice roots increased with an increase in Cr concentrations, while there was a negligible difference in the shoots. The comparative analysis demonstrated that the vector strengths in roots under 'Pro+Cr(VI)' treatments were higher than 'Cr(VI)' treatments, suggesting that Pro improved Gly II activity more efficiently to reduce MG content in roots. Calculation of the gene expression variation factors (GEFs) indicated a positive effect of Pro application on the expression of Gly I and Gly ll-related genes, wherein a stronger impact was in roots than the shoots. Together, the vector analysis and gene expression data reveal that exogenous Pro chiefly improved Gly ll activity in rice roots which subsequently enhanced MG detoxification under Cr(VI) stress.

Effects of two food colorants on catalase and trypsin: Binding evidences from experimental and computational analysis.

Recently, growing concern has been paid to the toxicity of additives in food. The present study investigated the interaction of two commonly used food colorants, quinoline yellow (QY) and sunset yellow (SY), with catalase and trypsin under physiological conditions by fluorescence, isothermal titration calorimetry (ITC), ultraviolet-vis absorption, synchronous fluorescence techniques as well as molecular docking. Based on the fluorescence spectra and ITC data, both QY and SY could significantly quench the intrinsic fluorescence of catalase or trypsin spontaneously to form a moderate complex driven by different forces. Additionally, the thermodynamics results demonstrated QY bind more tightly to both catalase and trypsin than SY, suggesting QY poses more of a threat to two enzymes than SY. Furthermore, the binding of two colorants could not only lead to the conformational and microenvironmental alterations of both catalase and trypsin, but also inhibit the activity of two enzymes. This study provides an important reference for understanding the biological transportation of synthetic food colorants in vivo, and enhancing their risk assessment on food safety.

Individual and mutual effects of elevated carbon dioxide and temperature on salt and cadmium uptake and translocation by rice seedlings.

Plant kingdoms are facing increasingly harsh environmental challenges marked by the coexposure of salinity and pollution in the pedosphere and elevated CO2 and temperature in the atmosphere due to the rapid acceleration of industrialization and global climate change. In this study, we deployed a hydroponics-based experiment to explore the individual and mutual effects of different temperatures (low temperature, T1: 23°C; high temperature, T2: 27°C) and CO2 concentrations (ambient CO2: 360 ppm; medium CO2: 450 ppm; high CO2: 700 ppm) on the uptake and translocation of sodium chloride (NaCl, 0.0, 0.2, 0.6, and 1.1 g Na/L) and cadmium nitrate (Cd(NO3)2·4H2O, 0.0, 0.2, 1.8, and 5.4 mg Cd/L) by rice seedlings. The results indicated that Cd and Na exposure significantly (P< 0.05) inhibited plant growth, but T2 and medium/high CO2 alleviated the effects of Cd and Na on plant growth. Neither significant synergistic nor antagonistic effects of Cd and Na were observed, particularly not at T1 or high CO2. At increasing temperatures, relative growth rates increased despite higher concentrations of Cd and Na in both rice roots and shoots. Similarly, higher CO2 stimulated the growth rate but resulted in significantly lower concentrations of Na, while the Cd concentration was highest at medium CO2. Coexposure experiments suggested that the concentration of Cd in roots slightly declined with additional Na and more at T2. Overall, our preliminary study suggested that global climate change may alter the distribution of mineral and toxic elements in rice plants as well as the tolerance of the plants.

Merging the occurrence possibility into gene co-expression network deciphers the importance of exogenous 2-oxoglutarate in improving the growth of rice seedlings under thiocyanate stress.

Thiocyanate (SCN-) can find its way into cultivated fields, which might hamper the harmony in carbon and nitrogen metabolism (CNM) of plants, ebbing their quality and productivity. In the current study, we investigated the role of the exogenous application of 2-oxoglutarate (2-OG) in maintaining homeostasis of CNM in rice seedlings under SCN- stress. Results showed that SCN- exposure significantly repressed the gene expression and activities of CNM-related enzymes (e.g., phosphoenolpyruvate carboxylase, NADP-dependent isocitrate dehydrogenases, and isocitrate dehydrogenases) in rice seedlings, thereby reducing their relative growth rate (RGR). Exogenous application of 2-OG effectively mitigated the toxic effects of SCN- on rice seedlings, judged by the aforementioned parameters. The co-expression network analysis showed that genes activated in CNM pathways were categorized into four modules (Modules 1-4). In order to identify the key module activated in CNM in rice seedlings exposed to SCN-, the results from real-time quantitative PCR (RT-qPCR) tests were used to calculate the possibility of the occurrence of genes grouped in four different modules. Notably, Module 3 showed the highest occurrence probability, which is mainly related to N metabolism and 2-OG synthesis. We can conclude that exogenous application of 2-OG can modify the imbalance of CNM caused by SCN- exposure through regulating N metabolism and 2-OG synthesis in rice seedlings.

Exogenous proline activated an integrated response of NER and HR pathways to reduce DNA damage in rice seedlings under chromium stress.

The DNA damage induced by hexavalent chromium [Cr(VI)] pollutant causes a genotoxic effect on rice seedlings. Hereby, we examined the effects of exogenous proline (Pro) on the alleviation of DNA damage in rice seedlings under different effective concentrations of Cr(VI). Our results revealed that Cr(VI) stress induced reactive oxygen species (ROS), i.e., H2O2 and O2·- accumulation in rice seedlings, repressed genes expression activated in the homologous recombination (HR) and nucleotide excision repair (NER) pathways, and caused DNA damage. Exogenous application of Pro increased Cr accumulation in rice roots, but decreased Cr accumulation in rice shoots, wherein Pro application decreased ROS accumulation in both tissues of rice seedlings. The comet assays suggested that exogenous application of Pro significantly alleviated the DNA damage in rice seedlings during Cr(VI) treatments, judged by the Olive tail moment and tail DNA. Transcriptional assays revealed that exogenous Pro upregulated the expression level of genes associated with the HR and NER pathways and triggered coordinated actions of both repairing pathways to modulate DNA lesion in rice plants during exposure to Cr(VI). Calculations from gene expression variation factors showed that regulative effect of exogenous application of Pro on DNA repair pathways was highly activated at 2.0 mg Cr/L. The current study revealed that Cr(VI) affect rice plants and exogenous Pro rescue these effects by the activation of HR and NER pathways.

Spatial-temporal variations of proline and related amino acids reveal distinct nitrogenous utilization strategies in rice during detoxification of exogenous cyanide.

Cyanide (CN-) pollution in agricultural systems impairs amino acid metabolism in rice plants, hence decreasing their quality and yield. Meanwhile, little is known about the effects of CN- assimilation on the innate pool of proline (Pro) and its synthesis-related amino acids (Pro-AAs) in rice plants. In this study, a hydroponic experiment was carried out to investigate the effect of exogenous KCN on indigenous levels of Pro-AAs, i.e., Pro, glutamate (Glu), arginine (Arg), and ornithine (Orn) in rice seedlings fertilized with either nitrate (NO3-) or ammonium (NH4+) through the biochemical and RT-qPCR analysis. At the same KCN treatment concentration, the relative growth rate of NH4+-fed rice seedlings was considerably higher than that of NO3--fed rice seedlings, but the residual concentration of CN- in NH4+-fed rice tissues was lower than that of NO3--fed rice tissues. Based on the UPLC and stoichiometry molar ratio calculations, it is evident that the Glu pathway contributed significantly to Pro synthesis in rice under KCN + NO3- treatments; whereas the Orn pathway governed the synthesis of Pro in rice under KCN + NH4+ treatments. Moreover, transcriptional and bioinformatics analysis revealed that NH4+ fertilization resulted in spatial-temporal differences in the genetic response in rice tissue during detoxification of CN- compared with KCN + NO3- treatments. These findings suggested that the innate level of Pro serves as "a fishing float" to balance the flux between Pro and Pro-AAs in exogenous KCN-treated rice plants under different nitrogenous nutritional conditions.

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.

Transcriptional analysis of heavy metal P1B-ATPases (HMAs) elucidates competitive interaction in metal transport between cadmium and mineral elements in rice plants.

Cadmium (Cd) pollution has become a major threat to crop production and quality globally. The heavy metal P1B-ATPases (HMAs) play a crucial role in metal transport in plants. In the present study, we investigated the interaction in metal transport by HMAs between Cd and mineral elements in rice plants. Rice seedlings were treated with cadmium nitrate either in the nutrient solution ("Cd+M") or in the ultrapure water ("Cd-M"). Result showed that phytotoxicity of Cd to rice seedlings was evident from both Cd treatments, judged by relative growth rate (RGR), where more severe repression (p < 0.05) of RGR was observed in the "Cd-M" treatments than the "Cd+M" treatments. More Cd (p < 0.05) was accumulated in rice tissues from the "Cd-M" treatments than the "Cd+M" treatments, while there is a significant difference (p < 0.05) in distribution and translocation of mineral elements in rice tissues between the "Cd+M" and the "Cd-M" treatments. RT-qPCR analysis displayed that the expression patterns of HMAs related genes were quite different between "Cd+M" and "Cd-M" treatments, suggesting their different regulatory effects during the transport of Cd and mineral elements within rice plants. The competition in metal transport by HMAs mainly occurs between Cd and micro-elements of Zn and Cu in rice tissues during Cd exposure. Overall, this study provides new evidence to clarify the different translocation mechanisms of HMAs in metal transport between Cd and mineral elements in rice seedlings during Cd exposure.

Identification of the Key Genes Involved in Proline-Mediated Modification of Cell Wall Components in Rice Seedlings under Trivalent Chromium Exposure.

Chromium (Cr) toxicity exerts a detrimental effect on various physiological, biochemical, and molecular attributes of plants including the structure and functions of cell walls. On the other hand, the exogenous application of proline (Pro) is a beneficial strategy to overcome Cr toxicity. Therefore, it is a novel strategy to find the key genes associated with cell wall composition in rice under trivalent Cr with/without Pro application. A total of 203 genes were activated in the four cell wall biosynthesis pathways under chromium stress, namely cellulose (60), hemicellulose (57), lignin (35), and pectin (51). Based on the expression abundance of microarrays, the number of differentially expressed genes, and the expression level of genes, the lignin pathway was a crucial pathway in response to Cr treatments, followed by the cellulose pathway. Through the estimation of gene expression variation factors between 'Cr' and 'Cr+Pro' treatments, OsUGP1, OsBGLU24, OsBGLU29, OsBGLU33, OsBMY1, and OsBMY2 in the cellulose pathway; OsXTH9, OsXTH10, OsXTH16, OsGAUT3, OsGAUT19, OsGAUT28, OsXTH1, OsGAUT12, and OsGAUT21 in the hemicellulose pathway; OsPAL3, OsPAL3, OsPOX1, and OsPRX77 in the lignin pathway; and OsPME25, OsPGL27, OsPME26, OsPGL9, and OsPLL12 in the pectin pathway are the key genes involved in cell wall modification during Cr exposure with exogenous Pro application. The Pro-mediated activation of these genes could be crucial players in modifying the cell wall structure and composition of rice plants under Cr stress, which needs to be further clarified.

Transcriptome Analysis of Cyanide-Treated Rice Seedlings: Insights into Gene Functional Classifications.

Cyanide (CN-) pollution in agricultural systems can affect crop production. However, no data are available to describe the full picture of the responsive metabolic mechanisms of genes with known functions related to exogenous KCN exposure. In this study, we examined the transcriptome in rice seedlings exposed to potassium cyanide (KCN) using an Agilent 4×44K rice microarray to clarify the relationship between the differentially expressed genes (DEGs) and their function classifications. The number of DEGs (up-regulated genes/down-regulated genes) was 322/626 and 640/948 in the shoots and roots of CN--treated rice seedlings, respectively. Functional predication demonstrated that a total of 534 and 837 DEGs in shoots and roots were assigned to 22 COG categories. Four common categories listed on the top five COG classifications were detected in both rice tissues: signal transduction mechanisms, carbohydrate transport and metabolism, post-translational modification, protein turnover and chaperones, and transcription. A comparison of DEGs aligned to the same COG classification demonstrated that the majority of up-regulated/down-regulated DEGs in rice tissues were significantly different, suggesting that responsive and regulatory mechanisms are tissue specific in CN--treated rice seedlings. Additionally, fifteen DEGs were aligned to three different COG categories, implying their possible multiple functions in response to KCN stress. The results presented here provide insights into the novel responsive and regulatory mechanisms of KCN-responsive genes, and will serve as useful resources for further functional dissections of the physiological significance of specific genes activated in the exogenous KCN stress response 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.

Proline-mediated regulation on jasmonate signals repressed anthocyanin accumulation through the MYB-bHLH-WDR complex in rice under chromium exposure.

Toxic metal-induced overaccumulation of anthocyanin (ATH) in plants can oxidize proteins and break DNA. Herein, the role of exogenous proline (Pro) on the repression of ATH accumulation in rice seedlings during hexavalent chromium [Cr(VI)] exposure was studied. Results indicated that exogenous Pro-mediated regulation of jasmonate signals activated the MYB-bHLH-WDR complex to repress ATH accumulation in rice tissues under Cr(VI) stress. Biochemical and transcript analysis indicated that exogenous Pro promoted the synthesis of jasmonic acid (JA) and its molecularly active metabolite jasmonic acid isoleucine (JA-Ile) in rice tissues under Cr(VI) stress. Increment in the endogenous level of jasmonates positively triggered the expression of genes responsible for the JA signaling pathway and activated the MYB-bHLH-WDR complex, eventually repressing the glycosylation of anthocyanidin to form ATH in rice tissues. In conclusion, exogenous proline-mediated regulation on jasmonate signals was tissue-specific under Cr(VI) stress and a more positive effect was detected in shoots rather than roots.

Implications of the fate of hydrogen sulfide derived from assimilation of thiocyanate in rice plants.

Thiocyanate (SCN-) is a sulfur-containing pollutant, which is frequently detected in irrigation water and has negative effects on plant growth and crop yields. Uptake and assimilation of exogenous SCN- in rice plants was evident, in which two metabolic pathways, carbonyl sulfide (COS) and cyanate (CNO), are activated. Hydrogen sulfide (H2S) is an important concomitant derived from detoxification of exogenous SCN- in rice plants, which may cause coupling action on the endogenous source of H2S from sulfur metabolism. Since H2S has dual regulatory effects, the fate of H2S derived from assimilation of SCN- in plants is critical for clarifying the inclusiveness of H2S in various physiological activities. In fact, application of exogenous H2S not only positively changed the root phenotype traits of SCN--treated seedlings, but also effectively mitigated the toxic effects of SCN- in rice seedlings by stimulating the process of the PSII repair cycle. In this study, it is tempting to analyze and clarify the flux of the concomitant production of H2S from assimilation of exogenous SCN- into the innate pool, which may function in signaling regulation and other physiological processes in rice plants. This study would update our understanding of the fate of H2S derived from assimilation of SCN- in plants and provide new insights into the affirmative actions of H2S in direct proximity to SCN- exposure.

Mathematical quantification of interactive complexity of transcription factors involved in proline-mediated regulative strategies in Oryza sativa under chromium stress.

Involvement of transcription factor (TFs) in governing genes at transcription or post transcription level is known to have affirmative impact on plant physiological and morphological development, especially during environmental abuse. Application of exogenous proline (Pro) is one among the effective approaches to strengthen plant resistance against stresses. However, Pro-mediated regulative strategies of TFs in responses to the chromium (Cr) in rice plants through the gene interaction network are still not clear. In the current study, Pro-mediated interactive complexity of various TFs (i.e., MYB, NAC, WRKY, bHLH, and bZIP) under hexavalent chromium [Cr(VI)] was investigated using Agilent 4 × 44 K rice gene chip and gene interactive probability model (GIPM). Results showed that exogenous Pro had a negligible effect on Cr uptake in rice plants, while a small positive response in biomass accretion of rice seedlings was observed under Cr(VI)+Pro treatments which was to certain extend greater than single Cr(VI) treatments. Rice microarray analysis showed that Cr(VI) significantly (p < 0.05) repressed the expression of TFs in the rice roots and shoots, while the application of exogenous Pro significantly (p < 0.05) up-regulated the expression levels of some TFs in rice tissues. Mathematical modularization indicated that Pro-mediated interaction between MYB and NAC carried more weightage than other TFs in rice roots and shoots under Cr(VI) stress. Overall, our study provides convincing evidence to confirm a positive role of exogenous Pro on reducing the negative impact exerted by Cr(VI) on rice plants through regulating expression and interaction of different TFs.