Dobinin K Displays Antiplasmodial Activity through Disruption of Plasmodium falciparum Mitochondria and Generation of Reactive Oxygen Species.

Dobinin K is a novel eudesmane sesquiterpenoids compound isolated from the root of Dobinea delavayi and displays potential antiplasmodial activity in vivo. Here, we evaluate the antiplasmodial activity of dobinin K in vitro and study its acting mechanism. The antiplasmodial activity of dobinin K in vitro was evaluated by concentration-, time-dependent, and stage-specific parasite inhibition assay. The potential target of dobinin K on Plasmodium falciparum was predicted by transcriptome analysis. Apoptosis of P. falciparum was detected by Giemsa, Hoechst 33258, and TUNEL staining assay. The reactive oxygen species (ROS) level, oxygen consumption, and mitochondrial membrane potential of P. falciparum were assessed by DCFH-DA, R01, and JC-1 fluorescent dye, respectively. The effect of dobinin K on the mitochondrial electron transport chain (ETC) was investigated by enzyme activity analysis and the binding abilities of dobinin K with different enzymes were learned by molecular docking. Dobinin K inhibited the growth of P. falciparum in a concentration-, time-dependent, and stage-specific manner. The predicted mechanism of dobinin K was related to the redox system of P. falciparum. Dobinin K increased intracellular ROS levels of P. falciparum and induced their apoptosis. After dobinin K treatment, P. falciparum mitochondria lost their function, which was presented as decreased oxygen consumption and depolarization of the membrane potential. Among five dehydrogenases in P. falciparum ETC, dobinin K displayed the best inhibitory power on NDH2 activity. Our findings indicate that the antiplasmodial effect of dobinin K in vitro is mediated by the enhancement of the ROS level in P. falciparum and the disruption of its mitochondrial function.

Genome-wide identification and evolutionary analysis of RLKs involved in the response to aluminium stress in peanut.

BACKGROUND: As an important cash crop, the yield of peanut is influenced by soil acidification and pathogen infection. Receptor-like protein kinases play important roles in plant growth, development and stress responses. However, little is known about the number, location, structure, molecular phylogeny, and expression of RLKs in peanut, and no comprehensive analysis of RLKs in the Al stress response in peanuts have been reported. RESULTS: A total of 1311 AhRLKs were identified from the peanut genome. The AhLRR-RLKs and AhLecRLKs were further divided into 24 and 35 subfamilies, respectively. The AhRLKs were randomly distributed across all 20 chromosomes in the peanut. Among these AhRLKs, 9.53% and 61.78% originated from tandem duplications and segmental duplications, respectively. The ka/ks ratios of 96.97% (96/99) of tandem duplication gene pairs and 98.78% (646/654) of segmental duplication gene pairs were less than 1. Among the tested tandem duplication clusters, there were 28 gene conversion events. Moreover, all total of 90 Al-responsive AhRLKs were identified by mining transcriptome data, and they were divided into 7 groups. Most of the Al-responsive AhRLKs that clustered together had similar motifs and evolutionarily conserved structures. The gene expression patterns of these genes in different tissues were further analysed, and tissue-specifically expressed genes, including 14 root-specific Al-responsive AhRLKs were found. In addition, all 90 Al-responsive AhRLKs which were distributed unevenly in the subfamilies of AhRLKs, showed different expression patterns between the two peanut varieties (Al-sensitive and Al-tolerant) under Al stress. CONCLUSIONS: In this study, we analysed the RLK gene family in the peanut genome. Segmental duplication events were the main driving force for AhRLK evolution, and most AhRLKs subject to purifying selection. A total of 90 genes were identified as Al-responsive AhRLKs, and the classification, conserved motifs, structures, tissue expression patterns and predicted functions of Al-responsive AhRLKs were further analysed and discussed, revealing their putative roles. This study provides a better understanding of the structures and functions of AhRLKs and Al-responsive AhRLKs.

Crosstalk between melatonin and nitric oxide in plant development and stress responses.

Melatonin is widely involved in plant growth and stress responses as a master regulator. Melatonin treatment alters the levels of endogenous nitric oxide (NO) and NO affects endogenous melatonin content. Melatonin and NO may induce various plant physiological behavior through interaction mechanism. However, the interactions between melatonin and NO in plants are largely unknown. The review presented the metabolism of endogenous melatonin and NO and their relationship in plants. The interactions between melatonin and NO in plant growth and development and responses to environmental stress were summarized. The molecular mechanisms of interaction between melatonin and NO in plants were also proposed.

Genome-wide analysis of the MATE gene family in potato.

The multidrug and toxic compound extrusion (MATE) protein family is a newly discovered family of secondary transporters that extrude metabolic waste and a variety of antibiotics out of the cell using an electrochemical gradient of H+ or Na+ across the membrane. The main function of MATE gene family is to participate in the process of plant detoxification and morphogenesis. The genome-wide analysis of the MATE genes in potato genome was conducted. At least 48 genes were initially identified and classified into six subfamilies. The chromosomal localization of MATE gene family showed that they could be distributed on 11 chromosomes except chromosome 9. The number of amino acids is 145-616, the molecular weight of proteins is 15.96-66.13 KD, the isoelectric point is 4.97-9.17, and they were located on the endoplasmic reticulum with having 4-13 transmembrane segments. They contain only two parts of the exons and UTR without introns. Some members of the first subfamily of potato MATE gene family are clustered with At2g04070 and they may be related to the transport of toxic compounds such as alkaloids and heavy metal. The function of the members of the second subfamily may be similar to that of At3g23560, which is related to tetramethylammonium transport. Some members of the third subfamily are clustered with At3g59030 and they may be involved in the transport of flavonoids. The fifth subfamily may be related to the transport of iron ions. The function of the sixth subfamily may be similar to that of At4g39030, which is related to salicylic acid transport. There are three kinds of conserved motifs in potato MATE genes, including the motif 1, motif 2, and motif 3. Each motif has 50 amino acids. The number of each motif is different in the gene sequence, of which 45 MATE genes contain at least a motif, but there is no motif in ST0015301, ST0045283, and ST0082336. These results provide a reference for further research on the function of potato MATE genes.

Role of nitric oxide and hydrogen sulfide in plant aluminum tolerance.

As gasotransmitter, nitric oxide (NO) and hydrogen sulfide (H2S) are involved in the regulation of plant tolerance to abiotic stresses. Aluminum (Al) toxicity triggers synthesis of NO and H2S and seriously affects plant growth and productivity. Exogenous NO and H2S alleviate Al toxicity in plants. However, the physiological and molecular mechanisms of NO and H2S in alleviating Al toxicity are very scattered. In this review, the advances in the effects of Al on the content of endogenous NO and H2S and the mechanisms of exogenous NO and H2S in alleviating Al toxicity in plants are summarized and discussed. The signaling pathway for the roles of NO and H2S in alleviating Al toxicity is also proposed.

Nitric oxide suppresses aluminum-induced programmed cell death in peanut (Arachis hypoganea L.) root tips by improving mitochondrial physiological properties.

Aluminum (Al) stress alters nitric oxide (NO) and induces programmed cell death (PCD) in plants. Recent study has shown that NO inhibits Al-induced PCD. However, the mechanism of NO inhibiting Al-induced PCD has not been revealed yet. Here, we investigated the behavior of mitochondria during Al-induced PCD suppressed by NO in peanut. Seedlings of peanut was grown hydroponically in a controllable growth room. The mitochondrial physiological parameters were determined spectrophotometrically. The expression of AhANT and AhHsp70 was determined by quantitative RT-PCR. Al-induced cell death rapidly in peanut root tips is mitochondria-dependent PCD. There was a significantly negative relationship between PCD and mitochondrial NO/H2O2 level. Compared with Al treatment alone, the addition of NO donor sodium nitroprusside (SNP) increased the ratio of NO/H2O2, down-regulated AhANT expression and inhibited the opening of mitochondrial permeability transition pore (MPTP), up-regulated AhHsp70 expression and increased mitochondrial inner membrane potential (ΔΨm), reduced cytochrome c (Cyt c) release from mitochondria and caspase 3-like protease activity, while the effect of NO specific scavenger cPTIO supplement was opposite. NO suppresses Al-induce PCD in peanut root tips by improving mitochondrial physiological properties.

Regulation of gaseous signaling molecules on proline metabolism in plants.

Proline accumulation plays an important role in the response and adaptation of plants to abiotic stress. Gaseous signaling molecules such as nitric oxide (NO), carbon monoxide (CO), and hydrogen sulfide (H2S) are involved in complicated events of cell signaling. However, the regulatory mechanisms of gaseous signaling molecules on proline synthesis and degradation are still unclear. This review summarized the biosynthesis and degradation of proline. The role of gaseous signaling molecules and their cross-talk on proline metabolic regulation in plants was discussed along with the future perspectives.

The central role of hydrogen sulfide in plant responses to toxic metal stress.

With the increase of industrial wastes, sewage irrigation, chemical fertilizers and pesticides, metal contamination is increasingly serious. How to reduce the environmental risk has become a compelling problem in cultivated land. As a gaseous signal molecule, hydrogen sulfide (H2S) is involved in multiple plant responses to toxic metal stress. Metal stress rapidly triggers endogenous H2S production and exogenous H2S alleviates metal toxicity in plants. To elucidate the role of H2S in metal tolerance, the physiological and molecular mechanisms of H2S in alleviating metal toxicity is necessary to be reviewed. Here, the latest progress on endogenous H2S metabolism and the role of H2S in plant responses to toxic metal stress were summarized and discussed. The mechanisms of exogenous H2S in alleviating metal toxicity is proposed.

[Preparation and luminescent properties of light scattering fluorescent resin].

With the methods of compound-melting and high-temperature molding, PC/YAG:Ce fluorescent resin pieces were prepared. The transmittance of the prepared PC/YAG:Ce fluorescent resin in 500~800 nm reaches approximately 65% (0.71 mm thick, double sides polished). The fluorescent resin was characterized by XRD, SEM and PL. The X-ray diffraction (XRD) patterns demonstrated that the fluorescent resin was pure Y3Al5O12 phase. The scanning electron microscopy (SEM) images showed that YAG phosphor was distributed evenly in the fluorescent resin. The excitation spectra had a weak peak at 342 nm and a strong band at 448 nm. The broad emission peaks at about 532 nm can be attributed to 5d-->4f transition of Ce3+ ions. Decay curves for the fluorescence of PC/YAG:Ce fluorescent resin show that the lifetime of the fluorescent resin was 61.5 ns. The luminous efficacy of the white LED packaged by the fluorescent resin and blue LED chip was 81.12 lm · W(-1) at 100 mA. All results above of PC/YAG:Ce fluorescent resin indicate a promising fluorescent material for white LEDs.

[Preparation and optical properties of MgAl2O4/Ce:YAG transparent ceramics].

High-purity ultrafine MgAl2O4 powder was synthesized by metal-alkoxide method and calcining for 2-4 h. And then MgAl2O4/Ce:YAG transparent ceramics were fabricated by hot-pressed sintering and hot isostatic pressed sintering technique with YAG:Ce powder and MgAl2O4 powder. The transparent ceramics were characterized by XRD, SEM, EDS and fluorescence spectrometer, respectively. The results show that the crystal phase of the transparent ceramic was composed of MgAl2O4 and YAG,and the YAG phase dispersed well in the matrix of MgAl2O4. The excitation spectra had a weak band at 345 nm and a strong band at 475 nm. The broad emission peaks at about 533 nm were attributed to 5d-->4f transition of Ce3+ ions. Decay curves for the fluorescence of MgAl2O4/Ce:YAG transparent ceramic test show that the lifetime of the Ce:YAG glass ceramic was 59.74 ns. All results show that MgAl2O4/Ce:YAG transparent ceramic may be a promising fluorescent material for white LED applications.

Identification and functional characterization of the xyloglucan endotransglucosylase/hydrolase 32 (AhXTH32) in peanut during aluminum-induced programmed cell death.

The toxicity of aluminum (Al) in acidic soil is a prevalent problem and causes reduced crop yields. In the plant response to Al toxicity, programmed cell death (PCD) appears to be an important mechanism. The plant cell wall of crop roots is the predominant site targeted by Al. Here, studies of the capacities of different cell wall constituents (pectin, hemicellulose 1 {HC1} and HC2) to adsorb Al indicated that HC1 has the greater ability to bind Al. The activity of xyloglucan endotransglucosylase (XET) was significantly inhibited by Al in the Al-tolerant peanut cultivar '99-1507' compared to that in 'ZH 2' (Al-sensitive). Results from qPCR analysis suggested that the suppression of XET activity by Al was transcriptionally regulated and that xyloglucan endotransglucosylase/hydrolase 32 (AhXTH32) was the major contributor to these changes. The overexpression of AhXTH32 in Arabidopsis strongly inhibited root growth with a loss of viability in root cells and the occurrence of typical hallmarks of PCD, while largely opposite effects were observed after xth32 suppression. AhXTH32 contributed to the modulation XET and xyloglucan endohydrolase (XEH) activity in vivo. Taken together, our results demonstrate that Al-tolerant peanut cultivar root tips cell walls bind Al predominantly in the HC1 fraction, which results in the inhibition of AhXTH32, with consequences to root growth, Al sensitivity, the occurrence of PCD and the XET/XEH activity ratio.

Identification and functional characterization of REGULATORY PARTICLE NON-ATPASE 1a-like (AhRPN1a-like) in peanuts during aluminum-induced programmed cell death.

The toxicity of aluminum (Al) in acidic soil is a prevalent problem and causes reduced crop yields. In the plant response to Al toxicity, programmed cell death (PCD) appears to be one of the important mechanisms. However, the regulation of Al-induced PCD remains poorly understood. Here, we found that an uncharacterized protein REGULATORY PARTICLE NON-ATPASE 1a-like in peanut (AhRPN1a-like), located in the nucleus and cytoplasm, directly interacted with type I metacaspase in peanut (AhMC1). The overexpression of AhRPN1a-like in Arabidopsis strongly enhanced Al inhibition of root growth with a loss of root tip cell viability. Furthermore, in response to Al treatment, the VIGS knockdown line of AhRPN1a-like in peanut displayed decreased transcription of AhMC1, increased root growth, reduced Al-induced PCD and decreased 26S proteasomal activity. Taken together, these findings demonstrated that AhRPN1a-like interacted directly with AhMC1, and promotes the occurrence of Al-induced PCD via the 26S proteasome pathway, thereby reducing Al-resistance.

[Detection of Porphyromonas gingivalis rag genotypes in patients of chronic periodontitis with chronic obstructive pulmonary disease].

PURPOSE: To investigate the distribution of Porphyromonas gingivalis(P.g) rag genotypes in patients of chronic periodontitis with chronic obstructive pulmonary disease (COPD). METHODS: Thirty patients with chronic periodontitis and 30 patients with chronic periodontitis complicated with COPD were included. Saliva samples were collected from all subjects． The detection rate and rag genotype of P.g in saliva were detected by 16S rDNA polymerase chain reaction (PCR). SPSS 22.0 software package was used for statistical analysis. RESULTS: The positive rate of P.g was 76.67% in chronic periodontitis patients with COPD, and 63.33% in chronic periodontitis group, there was no significant difference between the two groups (P＞0.05). The detection rates of rag-1 genotype in the two groups were 70% and 30.77%, respectively, there was significant difference between the two groups(P＜0.05). The detection rates of rag-2, rag-3 and rag-4 in the two groups were not significantly different. CONCLUSIONS: Various rag genotypes can be found in patients of chronic periodontitis with COPD. Rag-1 might have more close correlation with the development of COPD.

Transcriptome analysis reveals significant difference in gene expression and pathways between two peanut cultivars under Al stress.

Aluminum (Al) toxicity is an important factor in limiting peanut growth on acidic soil. The molecular mechanisms underlying peanut responses to Al stress are largely unknown. In this study, we performed transcriptome analysis of the root tips (0-1 cm) of peanut cultivar ZH2 (Al-sensitive) and 99-1507 (Al-tolerant) respectively. Root tips of peanuts that treated with 100 µM Al for 8 h and 24 h were analyzed by RNA-Seq, and a total of 8,587 differentially expressed genes (DEGs) were identified. GO and KEGG pathway analysis excavated a group of important Al-responsive genes related to organic acid transport, metal cation transport, transcription regulation and programmed cell death (PCD). These homologs were promising targets to modulate Al tolerance in peanuts. It was found that the rapid transcriptomic response to Al stress in 99-1507 helped to activate effective Al tolerance mechanisms. Protein and protein interaction analysis indicated that MAPK signal transduction played important roles in the early response to Al stress in peanuts. Moreover, weighted correlation network analysis (WGCNA) identified a predicted EIL (EIN3-like) gene with greatly increased expression as an Al-associated gene, and revealed a link between ethylene signaling transduction and Al resistance related genes in peanut, which suggested the enhanced signal transduction mediated the rapid transcriptomic responses. Our results revealed key pathways and genes associated with Al stress, and improved the understanding of Al response in peanut.

Nitric oxide is a suppressor of aluminum-induced mitochondria and caspase-like protease-dependent programmed cell death in plants.

Aluminum (Al) promotes programmed cell death (PCD) in plants. Although a lot of knowledge about the mechanisms of Al tolerance has been learned, how Al-induced PCD is regulated by nitric oxide (NO) is poorly understood. Mitochondrion is the regulatory center for PCD. We found that Al reduced the level of mitochondrial NO/H2O2, promoted the opening of mitochondrial permeability transition pore, decreased mitochondrial inner membrane potential (∆ψm), and increased caspase-like protease activity. NO-specific scavenger cPTIO enhanced these effects that were reversed by NO donor sodium nitroprusside. Our data suggest that NO suppresses Al-induced PCD by improving mitochondrial physiological properties.

Nitric oxide acts as an antioxidant and inhibits programmed cell death induced by aluminum in the root tips of peanut (Arachis hypogaea L.).

Aluminum (Al) causes programmed cell death (PCD) in plants. Our previous studies have confirmed that nitric oxide (NO) inhibits Al-induced PCD in the root tips of peanut. However, the mechanism by which NO inhibits Al-induced PCD is unclear. Here the effects of NO on mitochondrial reactive oxygen species (ROS), malondialdehyde (MDA), activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX), expression of alternative oxidase (AhAOX) and cytochrome oxidase (AhCOX) were investigated in peanut (Arachis hypogaea L.) root tips treated with Al. The results showed that Al stress induced rapid accumulation of H2O2 and MDA and increased the ratio of SOD/APX. The up-regulation of AhAOX and AhCOX expressions was not enough to inhibit PCD occurrence. Sodium nitroprusside (SNP, a NO donor) decreased the ratio of SOD/APX and eliminated excess H2O2 and MDA, thereby inhibiting Al-induced PCD in the root tips of peanut. The expression of AhAOX and AhCOX was significantly enhanced in Al-induced PCD treated with SNP. But cPTIO (a NO specific scavenger) supply had the opposite effect. Taken together, these results suggested that lipid peroxidation induced by higher levels of H2O2 was an important cause of Al-induced PCD. NO-mediated inhibition of Al-induced PCD was related to a significant elimination of H2O2 accumulation by decreasing the ratio of SOD/APX and up-regulating the expression of AhAOX and AhCOX.

Analysis of Mitochondrial Markers of Programmed Cell Death.

Mitochondria play a crucial role in programmed cell death (PCD) in plants. In most cases of mitochondria-dependent PCD, cytochrome c (Cyt c) released from mitochondria due to the opening of mitochondrial permeability transition pore (MPTP) and the activation of caspase-like proteases. Here we describe the analytic methods of mitochondrial markers of PCD including mitochondria isolation, mitochondrial membrane permeability, mitochondrial inner membrane potential, Cytc release, ATP, and mitochondrial reactive oxygen species (ROS).

Nitric oxide inhibits aluminum-induced programmed cell death in peanut (Arachis hypoganea L.) root tips.

It had been reported that Aluminum (Al) stress altered nitric oxide (NO) concentration and induced programmed cell death (PCD) in plants. However, the relationship between NO and PCD occurrence under Al stress is unclear. The results showed that cell death induced by Al was significant negative correlation with the inhibition of Al on root elongation growth in peanut. AlCl3 at 100µmolL-1 induced DNA ladder, chromatin condensation, typical apoptotic chromatin condensation staining with DAPI, apoptosis related gene Hrs203j expression and caspase3-like protease activation in peanut root tip cells, and showed that Al-induced cell death in peanut root tip cells was a typical PCD. Exogenous NO donor sodium nitroprusside (SNP) at 200µmolL-1 inhibited Al-induced PCD occurrence, but NO specific scavenger cPTIO aggravated PCD production. It suggests that NO is a negative regulator of Al-induced PCD in peanut root tips.

Nitric Oxide Inhibits Al-Induced Programmed Cell Death in Root Tips of Peanut (Arachis hypogaea L.) by Affecting Physiological Properties of Antioxidants Systems and Cell Wall.

It has been reported that nitric oxide (NO) is a negative regulator of aluminum (Al)-induced programmed cell death (PCD) in peanut root tips. However, the inhibiting mechanism of NO on Al-induced PCD is unclear. In order to investigate the mechanism by which NO inhibits Al-induced PCD, the effects of co-treatment Al with the exogenous NO donor or the NO-specific scavenger on peanut root tips, the physiological properties of antioxidants systems and cell wall (CW) in root tip cells of NO inhibiting Al-induced PCD were studied with two peanut cultivars. The results showed that Al exposure induced endogenous NO accumulation, and endogenous NO burst increased antioxidant enzyme activity in response to Al stress. The addition of NO donor sodium nitroprusside (SNP) relieved Al-induced root elongation inhibition, cell death and Al adsorption in CW, as well as oxidative damage and ROS accumulation. Furthermore, co-treatment with the exogenous NO donor decreased MDA content, LOX activity and pectin methylesterase (PME) activity, increased xyloglucan endotransglucosylase (XET) activity and relative expression of the xyloglucan endotransglucosylase/hydrolase (XTH-32) gene. Taken together, exogenous NO alleviated Al-induced PCD by inhibiting Al adsorption in CW, enhancing antioxidant defense and reducing peroxidation of membrane lipids, alleviating the inhibition of Al on root elongation by maintaining the extensibility of CW, decreasing PME activity, and increasing XET activity and relative XTH-32 expression of CW.

[Effects of sodium nitroprusside on mitochondrial function of rye and wheat root tip under aluminum stress].

Sodium nitroprusside (SNP) could ameliorate the inhibition effect of Al on root growth of rye (Secale cereale L. cv. King) and wheat (Triticum aestivum L. cv. Jinmai47). Respiratory rate, P/O, OPR (oxygenated phosphate rate), R(3) (oxygen consumption rate with ADP and substrate present), R(4) (oxygen consumption rate with substrate) and RCR (respiratory control ratio, R(3)/R(4)) of root tips from rye and wheat decreased, as well as the activities of H(+)-ATPase, H(+)-PPase, Na(+)-K(+)-ATPase, Ca(2+)-ATPase and Mg(2+)-ATPase, but they increased with SNP treatment. It showed that mitochondrial respiratory functions of root tips from rye and wheat were damaged, phosphorylation was un-coupled by Al, but that of rye was less than that of wheat. Rye has high Al-resistance ability than wheat. SNP is one of donor of NO, it is suggested that NO can ameliorate remarkably respiratory dysfunction resulted from Al stress, so that NO can ameliorate the inhibition effect of Al on plant growth.