Identification and expression analysis of MAPK cascade gene family in foxtail millet (Setaria italica).

The mitogen-activated protein kinase (MAPK) cascade pathway is a highly conserved plant cell signaling pathway that plays an important role in plant growth and development and stress response. Currently, MAPK cascade genes have been identified and reported in a variety of plants including Arabidopsis thaliana, Oryza sativa, and Triticum aestivum, but have not been identified in foxtail millet (Setaria italica). In this study, a total of 93 MAPK cascade genes, including 15 SiMAPKs, 10 SiMAPKKs and 68 SiMAPKKKs genes, were identified by genome-wide analysis of foxtail millet, and these genes were distributed on nine chromosomes of foxtail millet. Using phylogenetic analysis, we divided the SiMAPKs and SiMAPKKs into four subgroups, respectively, and the SiMAPKKKs into three subgroups (Raf, ZIK, and MEKK). Whole-genome duplication analysis revealed that there are 14 duplication pairs in the MAPK cascade family in foxtail millet, and they are expanded by segmental replication events. Results from quantitative real-time PCR (qRT-PCR) revealed that the expression levels of most SiMAPKs and SiMAPKKs were changed under both exogenous hormone and abiotic stress treatments, with SiMAPK3 and SiMAPKK4-2 being induced under almost all treatments, while the expression of SiMAPKK5 was repressed. In a nutshell, this study will shed some light on the evolution of MAPK cascade genes and the functional mechanisms underlying MAPK cascade genes in response to hormonal and abiotic stress signaling pathways in foxtail millet (Setaria italica).

Involvement of NO and Ca2+ in the enhancement of cold tolerance induced by melatonin in winter turnip rape (Brassica rapa L.).

As a multifunctional phytohormone, melatonin (Mel) plays pivotal roles in plant responses to multiple stresses. However, its mechanism of action remains elusive. In the present study, we evaluated the role of NO and Ca2+ signaling in Mel enhanced cold tolerance in winter turnip rape. The results showed that the NO content and concentration of intracellular free Ca2+ ([Ca2+]cyt) increased by 35.42% and 30.87%, respectively, in the leaves of rape seedlings exposed to cold stress. Compared with those of the seedlings in cold stress alone, the NO content and concentration of [Ca2+]cyt in rape seedlings pretreated with Mel increased further. In addition, the Mel-mediated improvement of cold tolerance was inhibited by L-NAME (a NO synthase inhibitor), tungstate (a nitrate reductase inhibitor), LaCl3 (a Ca2+ channel blocker), and EGTA (a Ca2+ chelator), and this finding was mainly reflected in the increase in ROS content and the decrease in osmoregulatory capacity, photosynthetic efficiency and antioxidant enzyme activities, and expression levels of antioxidant enzyme genes. These findings suggest that NO and Ca2+ are necessary for Mel to improve cold tolerance and function synergistically downstream of Mel. Notably, the co-treatment of Mel with L-NAME, tungstate, LaCl3, or EGTA also inhibited the Mel-induced expression of MAPK3/6 under cold stress. In conclusion, NO and Ca2+ are involved in the enhancement of cold tolerance induced by Mel through activating the MAPK cascades in rape seedlings, and a crosstalk may exist between NO and Ca2+ signaling.

GR24-mediated enhancement of salt tolerance and roles of H2O2 and Ca2+ in regulating this enhancement in cucumber.

This study investigated the regulation of the exogenous strigolactone (SL) analog GR24 in enhancing the salt tolerance and the effects of calcium ion (Ca2+) and hydrogen peroxide (H2O2) on GR24's regulation effects in cucumber. The seedlings were sprayed with (1) distilled water (CK), (2) NaCl, (3) GR24, then NaCl, (4) GR24, then H2O2 scavenger, then NaCl, and (5) GR24, then Ca2+ blocker, then NaCl. The second true leaf was selected for biochemical assays. Under the salt stress, the exogenous GR24 maintained the ion balance, increased the activity of antioxidant enzymes, reduced the membrane lipid peroxidation, and increased the activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX), accompanied by a decrease in relative conductivity, an increase in the proline content, and elevated gene expression levels of antioxidant enzymes, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, calcium-dependent protein kinases (CDPKs), salt overly sensitive SOS1, CBL-interacting protein kinase 2 (CIPK2), and calcineurin B-like protein 3 (CBL3). Such protective effects triggered by GR24 were attenuated or almost abolished by ethylene glycol tetraacetic acid (EGTA), lanthanum chloride (LaCl3, Ca2+ channel blocker), diphenyleneiodonium (DPI, NADPH oxidase inhibitor), and dimethylthiourea (DMTU, hydroxyl radical scavenger). Our data suggest that exogenous GR24 is highly effective in alleviating salt-induced damages via modulating antioxidant capabilities and improving ionic homeostasis and osmotic balance and that H2O2 and Ca2+ are required for GR24-mediated enhancement of salt tolerance.

Exogenous sucrose influences KEA1 and KEA2 to regulate abscisic acid-mediated primary root growth in Arabidopsis.

Arabidopsis K+-efflux antiporter (KEA)1 and KEA2 are chloroplast inner envelope membrane K+/H+ antiporters that play an important role in plastid development and seedling growth. However, the function of KEA1 and KEA2 during early seedling development is poorly understood. In this work, we found that in Arabidopsis, KEA1 and KEA2 mediated primary root growth by regulating photosynthesis and the ABA signaling pathway. Phenotypic analyses revealed that in the absence of sucrose, the primary root length of the kea1kea2 mutant was significantly shorter than that of the wild-type Columbia-0 (Col-0) plant. However, this phenotype could be remedied by the external application of sucrose. Meanwhile, HPLC-MS/MS results showed that in sucrose-free medium, ABA accumulation in the kea1kea2 mutant was considerably lower than that in Col-0. Transcriptome analysis revealed that many key genes involved in ABA signals were repressed in the kea1kea2 mutant. We concluded that KEA1 and KEA2 deficiency not only affected photosynthesis but was also involved in primary root growth likely through an ABA-dependent manner. This study confirmed the new function of KEA1 and KEA2 in affecting primary root growth.

Small signaling molecules in plant response to cold stress.

Cold stress is one of the harsh environmental stresses that adversely affect plant growth and crop yields in the Qinghai-Tibet Plateau. However, plants have evolved mechanisms to overcome the impact of cold stress. Progress has been made in understanding how plants perceive and transduce low-temperature signals to tolerate cold stress. Small signaling molecules are crucial for cellular signal transduction by initiating the downstream signaling cascade that helps plants to respond to cold stress. These small signaling molecules include calcium, reactive oxygen species, nitric oxide, hydrogen sulfide, cyclic guanosine monophosphate, phosphatidic acid, and sphingolipids. The small signaling molecules are involved in many aspects of cellular and physiological functions, such as inducing gene expression and activating hormone signaling, resulting in upregulation of the antioxidant enzyme activities, osmoprotectant accumulation, malondialdehyde reduction, and photosynthesis improvement. We summarize our current understanding of the roles of the small signaling molecules in cold stress in plants, and highlight their crosstalk in cold signaling transduction. These discoveries help us understand how the plateau plants adapt to the severe alpine environment as well as to develop new crops tolerating cold stress in the Qinghai-Tibet Plateau.

Hydrogen sulfide and nitric oxide are involved in melatonin-induced salt tolerance in cucumber.

Hydrogen sulfide (H2S) is a novel gaseous signaling molecule in response to adversity stress. Melatonin (MT) is a multifunctional molecule that plays an important role in regulating plant stress resistance. However, the interactions between H2S and MT are still unknown. Therefore, the role of H2S in MT-induced salt tolerance was elucidated in this study by measuring the antioxidant defense system and photosynthetic characteristics of cucumber. In addition, the crosstalk among H2S, NO, and mitogen-activated protein kinase (MAPK) was investigated. Results showed that MT induced the production of H2S by significantly increasing the activity of L-/D-cysteine desulfhydrase, thereby regulating photosynthetic efficiency, antioxidant enzyme activity, and antioxidant enzyme gene expression in cucumber, thus alleviating reactive oxygen species burst by salt stress. In this process, the H2S and NO induced by MT were inhibited by NO scavenger (cPTIO) and H2S scavenger (HT) but not affected by MAPK inhibitor (U0126). Intriguingly, the expression of MAPK3/4/6/9 was inhibited by HT and cPTIO. These results suggested that H2S may act as downstream of MT, interact with NO and MAPK cascades, and jointly participate in the process of MT mitigating salt stress in cucumber. In addition, H2S and NO are upstream signaling molecules of the MAPK cascades.

Hydrogen peroxide is involved in strigolactone induced low temperature stress tolerance in rape seedlings (Brassica rapa L.).

Strigolactone (SL) is a plant hormone that can improve plant stress resistance by regulating physiological processes and gene expression. GR24 is a synthetic strigolactone, which can also be used as a plant growth regulator. In this paper, the effects of exogenous GR24 on the growth and development of rape (Brassica rapa L.) under low temperature (4 °C) were studied. The results showed that low temperature (4 °C) inhibited the growth of rape seedlings, and exogenous GR24 significantly alleviated the effect of low temperature stress on rape seedlings. Compared with 4 °C treatment, GR24 + 4 °C treatment can increase the cell viability, soluble protein and proline content, enhance antioxidant enzyme activity, inhibit the production of reactive oxygen species (ROS), improve photosynthesis, and reduce the relative conductivity of rape seedlings. Further research shows that H2O2 plays a central role in improving the cold resistance of rape seedlings by GR24. qRT-PCR results indicated that GR24 significantly increased the expression of genes. Mainly includes antioxidant enzyme genes, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase genes, mitogen-activated protein kinase (MAPK) genes and cold-regulated genes. These results indicate that GR24 improves the cold tolerance of plants by regulating the expression of related genes. RNA-seq analysis revealed that there were 152 differentially expressed genes (DGEs) in T (4 °C)_vs_ST (GR24 + 4 °C), including 100 up-regulated genes and 52 down-regulated genes. These DEGs play an important role in carbon metabolism pathway, oxidative phosphorylation pathway, antioxidant activity and photosynthesis pathways. We selected 11 differentially expressed genes for qRT-PCR verification, and the verification results were consistent with RNA-seq results.

Overexpression of Brassica campestris BcICE1 gene increases abiotic stress tolerance in tobacco.

In this study, a cDNA of ICE1 (inducer of CBF expression 1) gene, named BcICE1, was isolated from Brassica campestris 'Longyou 6'. The deduced protein has 499 amino acids containing a typical bHLH domain and is highly identical with AtICE1 (85.9%) from Arabidopsis thaliana. BcICE1 is located in the nucleus. The activities of SOD, CAT, POD, and APX and the transcriptional levels of SOD, CAT, and POD genes were higher in BcICE1-transgenic tobacco than in wild-type (WT) tobacco under cold stress. Compared with WT tobacco, proline, soluble sugar, and chlorophyll were enhanced, whereas malondialdehyde and relative conductivity were decreased in BcICE1-transgenic tobacco. The overexpression of BcICE1 in tobacco increased the expression of CBF1, CBF2, and other stress-related genes. Moreover, under salt and PEG (25%) stress, the activities of APX and GPX and content of soluble sugar and chlorophyll in BcICE1-transgenic tobacco were higher than those in WT tobacco. Our results suggest that BcICE1 plays an important role in the response to abiotic stress.

Novel naïve Bayes classification models for predicting the chemical Ames mutagenicity.

Prediction of drug candidates for mutagenicity is a regulatory requirement since mutagenic compounds could pose a toxic risk to humans. The aim of this investigation was to develop a novel prediction model of mutagenicity by using a naïve Bayes classifier. The established model was validated by the internal 5-fold cross validation and external test sets. For comparison, the recursive partitioning classifier prediction model was also established and other various reported prediction models of mutagenicity were collected. Among these methods, the prediction performance of naïve Bayes classifier established here displayed very well and stable, which yielded average overall prediction accuracies for the internal 5-fold cross validation of the training set and external test set I set were 89.1±0.4% and 77.3±1.5%, respectively. The concordance of the external test set II with 446 marketed drugs was 90.9±0.3%. In addition, four simple molecular descriptors (e.g., Apol, No. of H donors, Num-Rings and Wiener) related to mutagenicity and five representative substructures of mutagens (e.g., aromatic nitro, hydroxyl amine, nitroso, aromatic amine and N-methyl-N-methylenemethanaminum) produced by ECFP_14 fingerprints were identified. We hope the established naïve Bayes prediction model can be applied to risk assessment processes; and the obtained important information of mutagenic chemicals can guide the design of chemical libraries for hit and lead optimization.

In silico prediction of drug-induced myelotoxicity by using Naïve Bayes method.

Drug-induced myelotoxicity usually leads to decrease the production of platelets, red cells, and white cells. Thus, early identification and characterization of myelotoxicity hazard in drug development is very necessary. The purpose of this investigation was to develop a prediction model of drug-induced myelotoxicity by using a Naïve Bayes classifier. For comparison, other prediction models based on support vector machine and single-hidden-layer feed-forward neural network  methods were also established. Among all the prediction models, the Naïve Bayes classification model showed the best prediction performance, which offered an average overall prediction accuracy of [Formula: see text] for the training set and [Formula: see text] for the external test set. The significant contributions of this study are that we first developed a Naïve Bayes classification model of drug-induced myelotoxicity adverse effect using a larger scale dataset, which could be employed for the prediction of drug-induced myelotoxicity. In addition, several important molecular descriptors and substructures of myelotoxic compounds have been identified, which should be taken into consideration in the design of new candidate compounds to produce safer and more effective drugs, ultimately reducing the attrition rate in later stages of drug development.

Salinity induced the changes of root growth and antioxidative responses in two wheat cultivars.

This study aimed to investigate the inhibitory mechanism of root growth and to compare antioxidative responses in two wheat cultivars, drought-tolerant Ningchun and drought-sensitive Xihan, exposed to different NaCl concentrations. Ningchun exhibited lower germination rate, seedling growth, and lipid peroxidation than Xihan when exposed to salinity. The loss of cell viability was correlated with the inhibition of root growth induced by NaCl stress. Moreover, treatments with H2O2 scavenger dimethylthiourea and catalase (CAT) partly blocked salinity-induced negative effects on root growth and cell viability. Besides, the enhancement of superoxide radical and H2O2 levels, and the stimulation of CAT and diamine oxidase (DAO) as well as the inhibition of glutathione reductase (GR) were observed in two wheat roots treated with salinity. However, hydroxyl radical content increased only in Xihan roots under NaCl treatment, and the changes of soluble peroxidase (POD), ascorbate peroxidase (APX), superoxide dismutase (SOD), and cell-wall-bound POD activities were different in drought-tolerant Ningchun and drought-sensitive Xihan exposed to different NaCl concentrations. In conclusion, salinity might induce the loss of cell viability via a pathway associated with extracellular H2O2 generation, which was the primary reason leading to the inhibition of root growth in two wheat cultivars. Here, it was also suggested that increased H2O2 accumulation in the roots of drought-tolerant Ningchun might be due to decreased POD and GR activities as well as enhanced cell-wall-bound POD and DAO ones, while the inhibition of APX and GR as well as the stimulation of SOD and DAO was responsible for the elevation of H2O2 level in drought-sensitive Xihan roots.

Expression and signal regulation of the alternative oxidase genes under abiotic stresses.

Plants in their natural environment frequently face various abiotic stresses, such as drought, high salinity, and chilling. Plant mitochondria contain an alternative oxidase (AOX), which is encoded by a small family of nuclear genes. AOX genes have been shown to be highly responsive to abiotic stresses. Using transgenic plants with varying levels of AOX expression, it has been confirmed that AOX genes are important for abiotic stress tolerance. Although the roles of AOX under abiotic stresses have been extensively studied and there are several excellent reviews on this topic, the differential expression patterns of the AOX gene family members and the signal regulation of AOX gene(s) under abiotic stresses have not been extensively summarized. Here, we review and discuss the current progress of these two important issues.

Cell death of rice roots under salt stress may be mediated by cyanide-resistant respiration.

Treatment with solutions containing high concentrations of NaCl (200 or 300 mM) induced cell death in rice (Oryza sativa L.) roots, as well as the application of exogenous hydrogen peroxide (H2O2). Moreover, the pretreatment with dimethylthiourea (DMTU), a scavenger of H2O2, partially alleviated the root cell death induced by 200 mM NaCl. These observations suggest that the cell death of rice roots under high salt stress is linked to H2O2 accumulation in vivo. NaCl stress increased the level of cyanide-resistant respiration to some extent and enhanced the transcript levels of the alternative oxidase (AOX) genes AOX1a and AOX1b in rice roots. High-salt-stressed (200 mM NaCl) rice roots pretreated with 1 mM salicylhydroxamic acid (SHAM), a specific inhibitor of alternative oxidase, exhibited higher levels of cell death and H2O2 production than roots subjected to either 200 mM NaCl stress or SHAM treatment alone. These results suggest that cyanide-resistant respiration could play a role in mediating root cell death under high salt stress. Furthermore, this function of cyanide-resistant respiration could relate to its ability to reduce the generation of H2O2.

Responses of seedling growth and antioxidant activity to excess iron and copper in Triticum aestivum L.

The purpose of this study was to analyze phytotoxicity mechanism involved in root growth and to compare physiological changes in the leaves of wheat seedlings exposed to short term iron (Fe) and copper (Cu) stresses (0, 100, 300 and 500µM). All applied Fe or Cu concentrations reduced root and shoot lengths, but seed germination was inhibited by Cu only at 500µM. Analyses using fluorescent dye 2',7'-dichlorodihydrofluorescein diacetate indicated enhanced H(2)O(2) levels in seedling roots under Fe and Cu treatments. Cu stress at the same concentration induced a great reduction in cell viability and a strong damage on membrane lipid in the roots with respect to Fe treatment. Significant increases in the total chlorophyll (chl) content including chl a and chl b were observed in response to higher Fe concentrations, whereas the highest Cu concentration (500µM) led to significant decreases in the total chl content including chl a. Additionally, leaf peroxidase (POD) and ascorbate peroxidase (APX) were stimulated by Fe stress, but the highest Fe concentration exhibited inhibitory effect on leaf APX activity. In contrast, copper treatment resulted in an elevation in leaf catalase and POD activities. Therefore, H(2)O(2) content in the leaves associated with copper was significantly lower than that with iron at the same concentration.

Zinc induced phytotoxicity mechanism involved in root growth of Triticum aestivum L.

This study investigated the inhibition mechanism of root growth in wheat seedlings when exposed to different zinc (Zn) concentrations. All applied Zn concentration did not affect seed germination, but reduced root length; in contrast, only Zn at 3mM inhibited significantly the growth of shoot. The loss of cell viability and the significant increases of lignification as well as the increases of hydrogen peroxide (H(2)O(2)), superoxide radical (O(2)(-)) and malondialdehyde levels were observed in the root tissue exposed to Zn treatment. And also, Zn stress led to the inhibition of cell-wall bound peroxidase. Moreover, NADPH oxidase inhibitor diphenylene iodonium could block greatly the elevation of O(2)(-) generation in Zn-treated roots. Therefore, the increased H(2)O(2) generation was dependent on the extracellular O(2)(-) production derived from plasma membrane NADPH oxidase. In addition, the loss of cell viability and the significant increases of lignification in response to the highest Zn concentration may be associated with the remarkable reduction of root growth in wheat seedlings.

Protective mechanism of desiccation tolerance in Reaumuria soongorica: leaf abscission and sucrose accumulation in the stem.

Reaumuria soongorica (Pall.) Maxim., a perennial semi-shrub, is widely found in semi-arid areas in northwestern China and can survive severe desiccation of its vegetative organs. In order to study the protective mechanism of desiccation tolerance in R. soongorica, diurnal patterns of net photosynthetic rate (Pn), water use efficiency (WUE) and chlorophyll fluorescence parameters of Photosystem II (PSII), and sugar content in the source leaf and stem were investigated in 6-year-old plants during progressive soil drought imposed by the cessation of watering. The results showed that R. soongorica was characterized by very low leaf water potential, high WUE, photosynthesis and high accumulation of sucrose in the stem and leaf abscission under desiccation. The maximum Pn increased at first and then declined during drought, but intrinsic WUE increased remarkably in the morning with increasing drought stress. The maximal photochemical efficiency of PSII (Fv/Fm) and the quantum efficiency of noncyclic electric transport of PSII (phiPSII) decreased significantly under water stress and exhibited an obvious phenomenon of photoinhibition at noon. Drought stressed plants maintained a higher capacity of dissipation of the excitation energy (measured as NPQ) with the increasing intensity of stress. Conditions of progressive drought promoted sucrose and starch accumulation in the stems but not in the leaves. However, when leaf water potential was less than -21.3 MPa, the plant leaves died and then abscised. But the stem photosynthesis remained and, afterward the plants entered the dormant state. Upon rewatering, the shoots reactivated and the plants developed new leaves. Therefore, R. soongorica has the ability to reduce water loss through leaf abscission and maintain the vigor of the stem cells to survive desiccation.

Effect of lanthanum ions (La3+) on ferritin-regulated antioxidant process under PEG stress.

The physiological effects of lanthanum(III) ions on the ferritin-regulated antioxidant process were studied in wheat (Triticum aestivum L.) seedlings under polyethylene glycol (PEG) stress. Treatment with 0.1 mM La3+ resulted in increased levels of chlorophyll, carotenoid, proline, ascorbate, and reduced glutathione. The activities of superoxide dismutase, catalase, ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, glutathione reductase, and peroxidase were also increased after La3+ treatment. Treatment with La3+ seems to enhance the capacity of the reactive oxygen species scavenging system, affect the Fe2+ and Fe3+ electron-transfer process in ferritin, and restrain the formation of hydroxyl radical (OH.), alleviating the oxidative damage induced by PEG stress.

Molecular cloning and characterization of a novel MAP kinase gene in Chorispora bungeana.

Chorispora bungeana Fisch. and C.A. Mey (Chorispora bungeana) is a rare alpine subnival plant species that is highly capable of resisting freezing environment. Since it is a stress-tolerant plant, we investigated the participation of mitogen-activated protein kinases (MAPKs) as possible mediators of abiotic stresses. We have isolated from Chorispora bungeana a new MAPK cDNA CbMAPK3 which encodes a 369 amino-acid protein with moderate to high nucleotide sequence similarity to previously reported plant MAPK genes. CbMAPK3 contains all 11 of the MAPK conserved subdomains and the phosphorylation motif TEY. The transcripts of CbMAPK3 were detected and no tissue-specific expression were observed in both roots and leaves, The transcripts of CbMAPK3 accumulated highly and rapidly when Chorispora bungeana treated with cold (4 and -4 degrees C), ABA and salinity stress. These results indicate that the CbMAPK3 may play an important role in response to environmental stresses.

Effects of enhanced ultraviolet-B radiation on algae and cyanobacteria.

This article provides an overview of existing literature on the ultraviolet-B (UV-B) radiation effects on algae and cyanobacteria. We report on the effects of UV-B radiation to the growth and development, biomass, sensitivity, photosynthetic pigments, UV-B absorbing compounds, photosynthesis, protein and DNA damage, enzyme activity, nitrogen fixation and assimilation of nitrogen, protective mechanisms of algae and cyanobacteria, the accommodation of algae and cyanobacteria to environmental stress, and the effects to ecology system. Many of the studies show the dramatic effects of UV-B radiation; but typically these studies were conducted under conditions with supplemental UV-B irradiance that was higher than would ever occur outside experimental conditions or natural condition. A few of the studies reviewed used experimental conditions and supplemental UV-B irradiance that approached realism. Enhanced UV-B generally decreased chlorophyll content, whereas it increased UV-B absorbing compounds in many algae. Decrease in photosynthesis, particularly at higher UV-B doses, was due to both direct (effect on photosystem) and indirect (decrease in pigments) effects. The decreases in chlorophyll pigments and photosynthesis resulted in lower biomass. However, algae and cyanobacteria have evolved various avoidance and repair mechanisms to protect themselves against the damaging effects of UV radiation to acclimate to enhanced UV-B radiation. The review points to areas where further studies on the relationships among nitrogenase, Rubisco, antioxidase activity, signal, antioxidants, and free radicals under enhanced UV-B are needed to quantify the effects of UV-B radiation on algae and cyanobacteria. These studies are needed in order to develop dose response functions that can facilitate development of dynamic simulation models for use in UV-B and other environmental impact assessments.