Biochemical insights into proline metabolism and its contribution to the endurant cell wall structure under metal stress.

The cell wall serves as the primary barrier against the entry of heavy metal ions into cells. However, excessive accumulation of heavy metals within plants can lead to alterations in the spatial structure and physical properties of the cell wall, thereby affecting the capacity of plants to capture heavy metals. Proline (Pro) is involved in the synthesis of the cell wall, modulating the stability and integrity of its structure. Extensins, core proteins that maintain the cell wall structure, are proline/hydroxyproline-rich glycoproteins that contain the characteristic sequence Ser-[Pro]3-5. They act as intermediates in the regulation of biological processes such as cell wall synthesis, assembly, and signal transduction, typically forming a network structure of cell wall proteins through cross-linking with pectin. This network is essential for the self-assembly expansion of the plant cell wall and plays an indispensable role in cell wall stress signal transduction through its interaction with intracellular signalling molecules. However, the mechanisms by which Pro affects the synthesis of cell wall structural proteins, cell wall assembly, and the sensing of cell wall stress under heavy metal stress remain unclear. This review, from the perspectives of biochemistry and molecular biology, comprehensively elaborates on the impact of Pro and Pro-rich proteins on the structure and function of the cell wall. These findings emphasize the mechanism by which Pro enhances the ability of the cell wall to capture heavy metals, providing new research ideas for the use of genetic engineering to manipulate cell wall synthesis and repair, thereby reducing the phytotoxicity of heavy metals.

Elucidating the intricacies of the H2S signaling pathway in gasotransmitters: Highlighting the regulation of plant thiocyanate detoxification pathways.

In recent decades, there has been increasing interest in elucidating the role of sulfur-containing compounds in plant metabolism, particularly emphasizing their function as signaling molecules. Among these, thiocyanate (SCN-), a compound imbued with sulfur and nitrogen, has emerged as a significant environmental contaminant frequently detected in irrigation water. This compound is known for its potential to adversely impact plant growth and agricultural yield. Although adopting exogenous SCN- as a nitrogen source in plant cells has been the subject of thorough investigation, the fate of sulfur resulting from the assimilation of exogenous SCN- has not been fully explored. There is burgeoning curiosity in probing the fate of SCN- within plant systems, especially considering the possible generation of the gaseous signaling molecule, hydrogen sulfide (H2S) during the metabolism of SCN-. Notably, the endogenous synthesis of H2S occurs predominantly within chloroplasts, the cytosol, and mitochondria. In contrast, the production of H2S following the assimilation of exogenous SCN- is explicitly confined to chloroplasts and mitochondria. This phenomenon indicates complex interplay and communication among various subcellular organelles, influencing signal transduction and other vital physiological processes. This review, augmented by a small-scale experimental study, endeavors to provide insights into the functional characteristics of H2S signaling in plants subjected to SCN--stress. Furthermore, a comparative analysis of the occurrence and trajectory of endogenous H2S and H2S derived from SCN--assimilation within plant organisms was performed, providing a focused lens for a comprehensive examination of the multifaceted roles of H2S in rice plants. By delving into these dimensions, our objective is to enhance the understanding of the regulatory mechanisms employed by the gasotransmitter H2S in plant adaptations and responses to SCN--stress, yielding invaluable insights into strategies for plant resilience and adaptive capabilities.

Functional Analysis of the Major Pilin Proteins of Type IV Pili in Streptococcus sanguinis CGMH010.

The pil gene cluster for Type IV pilus (Tfp) biosynthesis is commonly present and highly conserved in Streptococcus sanguinis. Nevertheless, Tfp-mediated twitching motility is less common among strains, and the factors determining twitching activity are not fully understood. Here, we analyzed the functions of three major pilin proteins (PilA1, PilA2, and PilA3) in the assembly and activity of Tfp in motile S. sanguinis CGMH010. Using various recombinant pilA deletion strains, we found that Tfp composed of different PilA proteins varied morphologically and functionally. Among the three PilA proteins, PilA1 was most critical in the assembly of twitching-active Tfp, and recombinant strains expressing motility generated more structured biofilms under constant shearing forces compared to the non-motile recombinant strains. Although PilA1 and PilA3 shared 94% identity, PilA3 could not compensate for the loss of PilA1, suggesting that the nature of PilA proteins plays an essential role in twitching activity. The single deletion of individual pilA genes had little effect on the invasion of host endothelia by S. sanguinis CGMH010. In contrast, the deletion of all three pilA genes or pilT, encoding the retraction ATPase, abolished Tfp-mediated invasion. Tfp- and PilT-dependent invasion were also detected in the non-motile S. sanguinis SK36, and thus, the retraction of Tfp, but not active twitching, was found to be essential for invasion.

Prevalence of Type IV Pili-Mediated Twitching Motility in Streptococcus sanguinis Strains and Its Impact on Biofilm Formation and Host Adherence.

Type IV pili (Tfp) are known to mediate several biological activities, including surface-dependent twitching motility. Although a pil gene cluster for Tfp biosynthesis is found in all sequenced Streptococcus sanguinis strains, Tfp-mediated twitching motility is less commonly detected. Upon examining 81 clinical strains, 39 strains generated twitching zones on blood agar plates (BAP), while 27 strains displayed twitching on Todd-Hewitt (TH) agar. Although BAP appears to be more suitable for the development of twitching zones, 5 strains exhibited twitching motility only on TH agar, indicating that twitching motility is not only strain specific but also sensitive to growth media. Furthermore, different twitching phenotypes were observed in strains expressing comparable levels of pilT, encoding the retraction ATPase, suggesting that the twitching phenotype on agar plates is regulated by multiple factors. By using a PilT-null and a pilin protein-null derivative (CHW02) of twitching-active S. sanguinis CGMH010, we found that Tfp retraction was essential for biofilm stability. Further, biofilm growth was amplified in CHW02 in the absence of shearing force, indicating that S. sanguinis may utilize other ligands for biofilm formation in the absence of Tfp. Similar to SK36, Tfp from CGMH010 were required for colonization of host cells, but PilT only marginally affected adherence and only in the twitching-active strain. Taken together, the results suggest that Tfp participates in host cell adherence and that Tfp retraction facilitates biofilm stability. IMPORTANCE Although the gene clusters encoding Tfp are commonly present in Streptococcus sanguinis, not all strains express surface-dependent twitching motility on agar surfaces. Regardless of whether the Tfp could drive motility, Tfp can serve as a ligand for the colonization of host cells. Though many S. sanguinis strains lack twitching activity, motility can enhance biofilm stability in a twitching-active strain; thus, perhaps motility provides little or no advantage to the survival of bacteria within dental plaque. Rather, Tfp retraction could provide additional advantages for the bacteria to establish infections outside the oral cavity.

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.

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.

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.

Regulation of enzymatic and non-enzymatic antioxidants in rice seedlings against chromium stress through sodium hydrosulfide and sodium nitroprusside.

Hydrogen sulfide (H2S) and nitric oxide (NO) play a well-organized protective mechanism in coping with oxidative stress induced by toxic metals. However, the comparative effects of H2S and NO on enzymatic and non-enzymatic antioxidants in plants under Cr(III) stress have not been defined. In this study, we mathematically evaluate the importance of sodium hydrosulfide (NaHS) and sodium nitroprusside (SNP) on these two antioxidant systems in rice seedlings under Cr(III) stress. The results displayed that the optimal dose of NaHS and SNP was 100 µM and 75 µM, respectively, in rice tissues under Cr(III) stress, judging by the reactive oxygen species (ROS) levels in rice tissues. When rice seedlings were exposed to Cr(III) at concentrations of 3.57, 7.24, 26.52 mg Cr/l, Cr-induced ROS accumulation had a significant (p < 0.05) effect on the lipid peroxidation of cell membrane in rice tissues, and decreased the response of SOD, CAT, Pro, GSH, and AsA in rice tissues. Application of exogenous NaHS and SNP effectively (p < 0.05) alleviated the toxic effects of Cr(III) in rice seedlings by activating different antioxidants. A novel physiological response model suggested that the regulatory effect of NaHS and SNP on non-enzymatic antioxidants was stronger than that of enzymatic antioxidants. Moreover, NaHS-mediated response intensity of both enzymatic and non-enzymatic antioxidants was greater than that of SNP, suggesting the importance of utilizing NaHS over SNP as antioxidant regulators during detoxification of Cr(III) in rice seedlings.

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.

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.

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.

The importance of utilizing nitrate (NO3-) over ammonium (NH4+) as nitrogen source during detoxification of exogenous thiocyanate (SCN-) in Oryza sativa.

Thiocyanate (SCN-) is a nitrogen-containing pollutant, which can be involved in the nitrogen (N) cycle and interferes with plant growth. The current study highlights a new insight into the N (nitrate [NO3-] and ammonium [NH4+]) utilization ways in rice seedlings under SCN- exposure to clarify the interactive effect on uptake and assimilation between these N-containing chemicals. Phenotypically, relative growth rates (RGR) of NO3--fed seedlings were significantly higher than NH4+-fed rice seedlings at the same SCN- concentration. Both N fertilizations have no significant influence on SCN- content and its assimilation in rice seedlings. However, significant accumulation of NO3- and NH4+ were detected in shoots prior to roots under SCN- stress. Enzymatic assay and mRNA analysis showed that the carbonyl sulfide (COS) pathway of SCN- degradation occurred in both roots and shoots of NO3--fed seedlings but only evident in roots of NH4+-fed seedlings. Moreover, the effect of SCN- on the activity of nitrate reductase (NR), glutamine synthetase (GS), and glutamate synthase (GOGAT) was negligible in NO3--fed seedlings, while GOGAT activity was significantly inhibited in shoots of NH4+-fed seedlings. Nitrogen use efficiency (NUE) estimation provided positive evidence in utilizing NO3- over NH4+ as the main N source to support rice seedling growth during detoxification of exogenous SCN-. Overall, SCN- pollution has unexpectedly changed the rice preference for N source which shifted from NH4+ to NO3-, suggesting that the interactions of SCN- with different N sources in terms of uptake and assimilation in rice plants should not be overlooked, especially at the plant N nutritional level.

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.

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.

Involvement of ß-cyanoalanine synthase (ß-CAS) and sulfurtransferase (ST) in cyanide (CN-) assimilation in rice seedlings.

In spite of available information demonstrating the assimilation of cyanide (CN-) by ß-cyanoalanine synthase (ß-CAS) in plants, involvement of sulfurtransferase (ST) in CN- assimilation in rice plants is still undefined. In this study, a microcosmic hydroponic system was used to investigate the involvement of ß-cyanoalanine synthase (ß-CAS) and sulfurtransferase (ST) in the CN- assimilation in rice seedlings under the exposure of potassium cyanide (KCN) in presence or absence of 1-amino-cyclopropane-1-carboxylic acid (ACC). Our results indicated that the measurable thiocyanate (SCN-) was detected in both rice roots and shoots under KCN exposure, and the abundances of ST-related transcripts were up-regulated significantly (p < 0.05), suggesting that the ST pathway is involved in CN- assimilation in the rice plants. The application of exogenous ACC significantly (p < 0.05) decreased the accumulation of CN- and SCN- in rice tissues after KCN exposures, and also up-regulated the expression of ß-CAS and ST genes and their enzymatic activities, suggesting a positive interaction between aminocyclopropane-1-carboxylate oxidase (ACO), ß-CAS and ST in rice plants during the CN- assimilation. This is the first attempt to experimentally clarify the involvement of ST in CN- assimilation in rice seedlings.

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.