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.

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.

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.

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.

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.

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.

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 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.

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.

The role of carbon monoxide signaling in the responses of plants to abiotic stresses.

Whether carbon monoxide (CO) exerts toxic or protective effect is dependent on the concentration and location of CO in animals. Similarly, it has been increasingly evident that CO also is involved in diverse physiological processes in plants, from seed germination and dormancy to stomatal closure to regulation of multiple environmental stresses. In this review, we focus on CO synthesis and the role of CO in plant responses to abiotic stresses, such as salinity, drought, cadmium and mercury. In general, abiotic stresses induce CO production in plants. CO can alleviate oxidative damage by improving the activities of antioxidative enzymes and antioxidant metabolism. In addition, cross talk between CO signaling and other signaling molecules including nitric oxide (NO) and hydrogen peroxide (H(2)O(2)) also is discussed.

Reactive oxygen species burst induced by aluminum stress triggers mitochondria-dependent programmed cell death in peanut root tip cells.

Recent studies had certified that aluminum (Al) induced ROS production and programmed cell death (PCD) in higher plants. The relationship between ROS production and PCD occurrence under Al stress is uncovered. The results showed that root elongation inhibition and PCD occurrence was induced by 100 µM AlCl3. Al stress induced ROS burst, up-regulated Rboh and COX gene expression, increased mitochondrial permeability transition pore (MPTP) opening, decreased inner mitochondrial membrane potential (ΔΨm), released cytochrome c from mitochondria to cytoplasm, activated caspase 3-like protease activity. Exogenous H2O2 aggravated the changes caused by Al and accelerated PCD occurrence, but ROS scavenger CAT and AsA reversed the changes caused by Al and inhibited PCD production. A potential cascade of cellular events during Al induced PCD via mitochondria dependent pathway and the mechanism of ROS on regulating PCD induced by Al is proposed.

The diversity of nitric oxide function in plant responses to metal stress.

Nitric oxide (NO) emerges as signalling molecule, which is involved in diverse physiological processes in plants. High mobility metal interferes with NO signaling. The exogenous NO alleviates metal stress, whereas endogenous NO contributes to metal toxicity in plants. Owing to different cellular localization and concentration, NO may act as multifunctional regulator in plant responses to metal stress. It not only plays a crucial role in the regulation of gene expression, but serves as a long-distance signal. Through tight modulation of redox signaling, the integration among NO, reactive oxygen species and stress-related hormones in plants determines whether plants stimulate death pathway or activate survival signaling.

Mitochondrial alterations during Al-induced PCD in peanut root tips.

Previous study found there was a negative relationship between Al-induced PCD and Al-resistance in peanut. The present research was undertaken to verify whether mitochondria play a significant role in PCD induced by Al in peanut. The roots of Al-tolerant plants were found to exhibit more intensive root growth, while accumulating less Al³âº than Al-sensitive plants under Al treatment. The different enhancement of ROS production was observed in the mitochondria isolated from two peanut cultivars. The concentration of mitochondrial MDA in root tips increased after Al treatment, which was higher in Zhonghua 2 than in 99-1507. With the increase of Al concentration, mitochondrial Ca²âº concentration decreased, and Ca²âº concentration of Zhonghua 2 decreased faster than that of 99-1507. The opening of mitochondrial permeability transition pore was more extensively in mitochondria isolated from Zhonghua 2 than from 99-1507. The collapse of inner mitochondrial membrane potential (ΔΨm) was also observed with a release of Cytochrome c (Cyt c) from mitochondria, it was more obvious in Zhonghua 2 than in 99-1507 with Al concentration increasing. The results showed that mitochondrial membrane structure and function were damaged seriously in Al-induced PCD, the increase of mitochondrial antioxidant system activity decreased cellular damages under Al stress. To sum up, compared with Al-sensitive peanut cultivar, Al-tolerant peanut cultivar has less Al³âº absorption, mitochondrial ROS and membrane lipid peroxidation level, higher control of MPT opening, ΔΨm maintaining, Cty c release from mitochondria and mitochondrial respiratory functions so that it is not easy to produce PCD under Al stress.

Role of microRNAs in aluminum stress in plants.

Aluminum (Al) stress is a major factor limiting crop production. The primary symptom of Al toxicity is to inhibit root growth. Plant responses to Al require precise regulation of gene expression at transcriptional and post-transcriptional levels. MicroRNAs (miRNAs) are 20-23 nucleotides length non-coding RNAs, which promote the cleavage of target mRNAs. We have summarized some Al-responsive miRNAs identified, especially proposed the regulatory roles of miR319, miR390, miR393, miR319a.2, and miR398 in Al stress signaling network. The cross-talk between miRNAs and signaling pathways also has been discussed.

Aluminum induces rapidly mitochondria-dependent programmed cell death in Al-sensitive peanut root tips.

BACKGROUND: Although many studies suggested that aluminum (Al) induced programmed cell death (PCD) in plants, the mechanism of Al-induced PCD and its effects in Al tolerance is limited. This study was to investigate the mechanism and type of Al induced PCD and the relationship between PCD and Al tolerance. RESULTS: In this study, two genotypes of peanut 99-1507 (Al tolerant) and ZH2 (Al sensitive) were used to investigate Al-induced PCD. Peanut root growth inhibition induced by AlCl3 was concentration and time-dependent in two peanut varieties. AlCl3 at 100 µM could induce rapidly peanut root tip PCD involved in DNA cleavage, typical apoptotic chromatin condensation staining with DAPI, apoptosis related gene Hrs203j expression and cytochrome C (Cyt c) release from mitochondria to cytosol. Caspase3-like protease was activated by Al; it was higher in ZH2 than in 99-1507. Al increased the opening of mitochondrial permeability transition pore (MPTP), decreased inner membrane potential (ΔΨm) of mitochondria. Compared with the control, Al stress increased O2•- and H2O2 production in mitochondria. Reactive oxygen species (ROS) burst was produced at Al treatment for 4 h. CONCLUSIONS: Al-induced PCD is earlier and faster in Al-sensitive peanut cultivar than in Al-tolerant cultivar. There is a negative relationship between PCD and Al resistance. Mitochondria- dependence PCD was induced by Al and ROS was involved in this process. The mechanism can be explained by the model of acceleration of senescence under Al stress.

Aluminum-induced programmed cell death promoted by AhSAG, a senescence-associated gene in Arachis hypoganea L.

Programmed cell death (PCD) is a foundational cellular process in plant development and elimination of damaged cells under environmental stresses. In this study, Al induced PCD in two peanut (Arachis hypoganea L.) cultivars Zhonghua 2 (Al-sensitive) and 99-1507 (Al-tolerant) using DNA ladder, TUNEL detection and electron microscopy. The concentration of Al-induced PCD was lower in Zhonghua 2 than in 99-1507. AhSAG, a senescence-associated gene was isolated from cDNA library of Al-stressed peanut with PCD. Open reading frame (ORF) of AhSAG was 474bp, encoding a SAG protein composed of 157 amino acids. Compared to the control and the antisense transgenic tobacco plants, the fast development and blossom of the sense transgenic plants happened to promote senescence. The ability of Al tolerance in sense transgenic tobacco was lower than in antisense transgenic tobacco according to root elongation and Al content analysis. The expression of AhSAG-GFP was higher in sense transgenic tobacco than in antisense transgenic tobacco. Altogether, these results indicated that there was a negative relationship between Al-induced PCD and Al-resistance in peanut, and the AhSAG could induce or promote the occurrence of PCD in plants.

Interactions between nitric oxide and plant hormones in aluminum tolerance.

Nitric oxide (NO) is involved, together with plant hormones, in the adaptation to Al stress in plants. However, the mechanism by which NO and plant hormones interplay to improve Al tolerance are still unclear. We have recently shown that patterns of plant hormones alteration differ between rye and wheat under Al stress. NO may enhance Al tolerance by regulating hormonal equilibrium in plants, as a regulator of plant hormones signaling. In this paper, some unsolved issues are discussed based on recent studies and the complex network of NO and plant hormones in inducing Al tolerance of plants are proposed.

Nitric oxide signaling in aluminum stress in plants.

Nitric oxide (NO) is a ubiquitous signal molecule involved in multiple plant responses to environmental stress. In the recent years, the regulating role of NO on heavy metal toxicity in plants is realized increasingly, but knowledge of NO in alleviating aluminum (Al) toxicity is quite limited. In this article, NO homeostasis between its biosynthesis and elimination in plants is presented. Some genes involved in NO/Al network and their expressions are also introduced. Furthermore, the role of NO in Al toxicity and the functions in Al tolerance are discussed. It is proposed that Al toxicity may disrupt NO homeostasis, leading to endogenous NO concentration being lower than required for root elongation in plants. There are many evidences that pointed out that the exogenous NO treatments improve Al tolerance in plants through activating antioxidative capacity to eliminate reactive oxygen species. Most of the work with respect to NO regulating pathways and functions still has to be done in the future.