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Find video protocols related to scientific articles indexed in Pubmed.
Electrochemical, spectroscopic and theoretical studies of a simple bifunctional cobalt corrole catalyst for oxygen evolution and hydrogen production.
Phys Chem Chem Phys
PUBLISHED: 10-14-2014
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Six cobalt and manganese corrole complexes were synthesized and examined as single-site catalysts for water splitting. The simple cobalt corrole [Co(tpfc)(py)2] (1, tpfc = 5,10,15-tris(pentafluorophenyl)corrole, py = pyridine) catalyzed both water oxidation and proton reduction efficiently. By coating complex 1 onto indium tin oxide (ITO) electrodes, the turnover frequency for electrocatalytic water oxidation was 0.20 s(?1) at 1.4 V (vs. Ag/AgCl, pH = 7), and it was 1010 s(?1) for proton reduction at ?1.0 V (vs. Ag/AgCl, pH = 0.5). The stability of 1 for catalytic oxygen evolution and hydrogen production was evaluated by electrochemical, UV-vis and mass measurements, scanning electron microscope (SEM) and energy dispersive X-ray spectroscopy (EDX), which confirmed that 1 was the real molecular catalyst. Titration and UV-vis experiments showed that the pyridine group on Co dissociated at the beginning of catalysis, which was critical to subsequent activation of water. A proton-coupled electron transfer process was involved based on the pH dependence of the water oxidation reaction catalyzed by 1. As for manganese corroles 2–6, although their oxidizing powers were comparable to that of 1, they were not as stable as 1 and underwent decomposition at the electrode. Density functional theory (DFT) calculations indicated that water oxidation by 1 was feasible through a proposed catalytic cycle. The formation of an O–O bond was suggested to be the rate-determining step, and the calculated activation barrier of 18.1 kcal mol(?1) was in good agreement with that obtained from experiments.
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Melatonin regulates proteomic changes during leaf senescence in Malus hupehensis.
J. Pineal Res.
PUBLISHED: 09-12-2014
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Despite the relationship between melatonin and aging, the overall changes and regulation of proteome profiling by long-term melatonin exposure during leaf senescence is not well understood. In this study, leaf senescence in Malus hupehensis plants was delayed when exogenous melatonin was regularly applied to the roots for 2 months compared with natural leaf senescence. Proteins of samples 0 and 50 day for both treatments were extracted and labeled with TMT regents before being examined via NanoLC-MS/MS. The proteomics data showed that 622 and 309 proteins were altered by senescence and melatonin, respectively. Our GO analysis by Blast2GO revealed that most of the altered proteins that are involved in major metabolic processes exhibited hydrolase activity and were mainly located in the plastids. These proteins were classified into several senescence-related functional categories, including degradation of macromolecules, redox and stress responses, transport, photosynthesis, development, and other regulatory proteins. We found that melatonin treatment led to the downregulation of proteins that are normally upregulated during senescence. The melatonin-related delay in senescence might have occurred due to the altering of proteins involved in processes associated with senescence. And as well, there are many unknown regulatory proteins possibly being involved in the melatonin's function. This study is the first to demonstrate changes at the proteome level in response to exogenous melatonin in plants. Our findings provide a set of informative and fundamental data about the role of melatonin in apple leaf senescence.
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Photoprotection mechanism in the 'Fuji' apple peel at different levels of photooxidative sunburn.
Physiol Plant
PUBLISHED: 09-03-2014
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The xanthophyll cycle, flavonoid metabolism, the antioxidant system and the production of active oxygen species were analyzed in the peel of 'Fuji' apples re-exposed to sunlight after extended periods of fruit bagging treatment, resulting in different levels of photooxidative sunburn. After re-exposing bagged fruits to sunlight, the production of active oxygen species and the photoprotective capacity in apple peels were both significantly enhanced. As sunburn severity increased, the concentration of hydrogen peroxide increased, while xanthophyll cycle pool size decreased. For the key genes involved in flavonoid synthesis, expressions of MdMYB10 and MdPAL were upregulated, whereas the expressions of MdCHS, MdANS, MdFLS and MdUFGT were downregulated in sunburnt fruit peel. Correspondingly, concentrations of both quercetin-3-glycoside and cyanidin-3-galactoside decreased. Total ascorbate concentrations decreased as sunburn severity increased, with the decrease being faster for oxidized than for reduced ascorbate. Transcription levels of MdGMP, MdGME, MdGGP, MdGPP, MdGalDH and MdGalLDH, the genes involved in ascorbate synthesis, were similar in non-sunburnt and sunburnt fruit peels, whereas activities of l-galactose dehydrogenase and l-galactono-1,4-lactone dehydrogenase decreased in severely sunburnt peel. Although activities of superoxide dismutase and ascorbate peroxidase increased, the activities of monodehydroascorbate reductase, dehydroascorbate reductase and glutathione reductase decreased as sunburn severity increased. In summary, the occurrence of photooxidative sunburn in 'Fuji' apple peel is closely associated with a relatively lower xanthophyll cycle pool size, reduced levels of ascorbate reduction and synthesis and reduced flavonoid synthesis. Our data are consistent with the idea that ascorbate plays a key role in protecting apple fruit from photooxidative sunburn.
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Dopamine alleviates salt-induced stress in Malus hupehensis.
Physiol Plant
PUBLISHED: 08-25-2014
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Dopamine mediates many physiological processes in plants. We investigated its role in regulating growth, ion homeostasis and the response to salinity in Malus hupehensis Rehd. Both hydroponics and field-pot experiments were conducted under saline conditions. Salt-stressed plants had reduced growth and a marked decline in their net photosynthetic rates, values for Fv /Fm and chlorophyll contents. However, pretreatment with 100 or 200??M dopamine significantly alleviated this inhibition and enabled plants to maintain their photosynthetic capacity. In addition to changing stomatal behavior, supplementation with dopamine positively influenced the uptake of K, N, P, S, Cu and Mn ions but had an inhibitory effect on Na and Cl uptake, the balance of which is responsible for managing the response to salinity by Malus plants. Dopamine pretreatment also controlled the burst of hydrogen peroxide, possibly through direct scavenging and by enhancing the activities of antioxidative enzymes and the capacity of the ascorbate-glutathione cycle. We also investigated whether dopamine might regulate salt overly sensitive pathway genes under salinity. Here, MdHKT1, MdNHX1 and MdSOS1 were greatly upregulated in roots and leaves, which possibly contributed to the maintenance of ion homeostasis and, thus, improved salinity resistance in plants exposed earlier to exogenous dopamine. These results support our conclusion that dopamine alleviates salt-induced stress not only at the level of antioxidant defense but also by regulating other mechanisms of ion homeostasis.
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Cloning and expression of two 9-cis-epoxycarotenoid dioxygenase genes during fruit development and under stress conditions from Malus.
Mol. Biol. Rep.
PUBLISHED: 06-25-2014
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There is now biochemical and genetic evidence that oxidative cleavage of cis-epoxycarotenoids by 9-cis-epoxycarotenoid dioxygenase (NCED) is the critical step in the regulation of abscisic acid (ABA) synthesis in higher plants. To understand the expression characteristics of NCED during ABA biosynthesis in apple (Malus), two NCED genes cDNA sequence were cloned from Malus prunifolia using RT-PCR techniques, named MpNCED1 and MpNCED2. The two cDNA sequences have full-length open reading frame, encoding a polypeptide of 607 and 614 amino acids, respectively. Sequences analysis showed that the deduced two apple NCED proteins were highly homologous to other NCED proteins from different plant species. Real-time PCR analysis revealed MpNCED2 were expressed continuously during the whole period of apple fruit development with the pattern of "higher-low-highest", while the expression of MpNCED1 clearly declined to a steady low level in the mid-later period of fruit development. Expression of the MpNCED2 increased under the drought stress, high temperature and low temperature strongly and rapidly, whereas expression of the MpNCED1 was detected in response to temperature stress, but did not detected under drought stress. These results revealed that MpNCED1 and MpNCED2 may play different roles in regulation of the ABA biosynthesis in fruit development and various stresses response.
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Genome-wide identification and expression profiling of the SnRK2 gene family in Malus prunifolia.
Gene
PUBLISHED: 05-25-2014
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Sucrose non-fermenting-1-related protein kinase 2 (SnRK2) constitutes a small plant-specific serine/threonine kinase family with essential roles in the abscisic acid (ABA) signal pathway and in responses to osmotic stress. Although a genome-wide analysis of this family has been conducted in some species, little is known about SnRK2 genes in apple (Malus domestica). We identified 14 putative sequences encoding 12 deduced SnRK2 proteins within the apple genome. Gene chromosomal location and synteny analysis of the apple SnRK2 genes indicated that tandem and segmental duplications have likely contributed to the expansion and evolution of these genes. All 12 full-length coding sequences were confirmed by cloning from Malus prunifolia. The gene structure and motif compositions of the apple SnRK2 genes were analyzed. Phylogenetic analysis showed that MpSnRK2s could be classified into four groups. Profiling of these genes presented differential patterns of expression in various tissues. Under stress conditions, transcript levels for some family members were up-regulated in the leaves in response to drought, salinity, or ABA treatments. This suggested their possible roles in plant response to abiotic stress. Our findings provide essential information about SnRK2 genes in apple and will contribute to further functional dissection of this gene family.
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Leaf micromorphology and sugar may contribute to differences in drought tolerance for two apple cultivars.
Plant Physiol. Biochem.
PUBLISHED: 04-14-2014
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Apple trees (Malus domestica L.) are often exposed to severe water stress in the summertime. We determined how levels of nonstructural carbohydrates (NC) changed in the leaves of two cultivars - drought-tolerant 'QinGuan' and drought-sensitive 'NaganoFuji' - when plants were subjected for 10 d to a sudden water deficit. Photosynthetic performance was investigated in terms of gas exchange and antioxidant enzyme activity. Two separate irrigation scenarios were tested with one-year-old plants grafted onto Malus hupehensis (Pamp.) Rehd. rootstock: 1) water applied to 75% field capacity (FC) (Control, CK) and 2) irrigation withheld to achieve <75% FC ranging from 30 to 75% (Treatment, T). At Days 0, 1, 2, 4, 6, and 10 of the experiment, we recorded net CO2 assimilation, stomatal conductance, leaf transpiration rates, and intercellular CO2 concentrations. All of those parameters showed decreases in plants from both stress scenarios, although those declines were not as dramatic in 'QinGuan'. The photosynthetic rate reduced primarily because of stomatal closure. In both cultivars, water stress induced the accumulation of NC, especially sorbitol, suggesting that this polyol has a role in osmoregulation. The rise in sorbitol levels was much steeper in 'QinGuan'. Likewise, the interaction between superoxide dismutase and peroxidase activities varied between cultivars. In 'QinGuan', the malondialdehyde concentration was much lower and more closely correlated with a high Suc/NC ratio when compared with 'NaganoFuji'. Scanning electron microscopy revealed a drought-resisting apparent structure in 'QinGuan'. Overall, our results from both external and internal examinations demonstrated that 'QinGuan' is more drought-tolerant than 'NaganoFuji'.
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Genome-wide identification and expression profiling of the cystatin gene family in apple (Malus × domestica Borkh.).
Plant Physiol. Biochem.
PUBLISHED: 03-11-2014
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Cystatins or phytocystatins (PhyCys) comprise a family of plant-specific inhibitors of cysteine proteinases. Such inhibitors are thought to be involved in the regulation of several endogenous processes as well as defense against biotic or abiotic stresses. However, information about this family is limited in apple. We identified 26 PhyCys genes within the entire apple genome. They were clustered into three distinct groups distributed across several chromosomes. All of their putative proteins contained one or two typical cystatin domains, which shared the characteristic motifs of PhyCys. Eight selected genes displayed differential expression patterns in various tissues. Moreover, their transcript levels were also up-regulated significantly in leaves during maturation, senescence or in response to treatment with one or more abiotic stresses. Our results indicated that members of this family may function in tissue development, leaf senescence, and adaptation to adverse environments in apple.
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Reactive oxygen species produced via plasma membrane NADPH oxidase regulate anthocyanin synthesis in apple peel.
Planta
PUBLISHED: 01-30-2014
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Solar ultraviolet irradiation regulates anthocyanin synthesis in apple peel by modulating the production of reactive oxygen species via plasma membrane NADPH oxidase instead of other pathways. The synthesis of anthocyanin in apple peels is dependent upon solar irradiation. Using 3-mm commercial glass to attenuate solar UV-A and UV-B light, we confirmed that solar UV irradiation regulated anthocyanin synthesis in apple peels after exposing previously bagged fruit to sunlight. During sunlight exposure, UV attenuation did not affect the expression of MdHY5, MdCOP1, or MdCRY2, but significantly lowered plasma membrane NADPH oxidase activity and superoxide anion concentrations. UV attenuation also reduced the expression levels of MdMYB10, MdPAL, MdCHS, MdF3H, MdDFR, MdANS and MdUFGT1, UDP-glycose:flavonoid 3-O-glycosyltransferase (UFGT) activity, and local concentrations of anthocyanin and quercetin-3-glycoside. In contrast, exogenous application of hydrogen peroxide could enhance anthocyanin and quercetin-3-glycoside synthesis. Xanthophyll cycle pool size on a chlorophyll basis was higher but its de-epoxidation was lower under direct sunlight irradiation than that under UV-attenuating conditions. This suggests that reactive oxygen species (ROS) produced in chloroplast are not major contributors to anthocyanin synthesis regulation. Inhibition of plasma membrane NADPH oxidase activity lowered the production of ROS through this mechanism, significantly inhibited the synthesis of anthocyanin, and increased the total production of ROS in apple peel under direct sunlight irradiation, suggesting that ROS produced via plasma membrane NADPH oxidase regulates anthocyanin synthesis. In summary, solar UV irradiation regulated anthocyanin synthesis in apple peels by modulating the production of ROS via plasma membrane NADPH oxidase.
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Screening and identification of resistance related proteins from apple leaves inoculated with Marssonina coronaria (EII. & J. J. Davis).
Proteome Sci
PUBLISHED: 01-27-2014
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Apple, an invaluable fruit crop worldwide, is often prone to infection by pathogenic fungi. Identification of potentially resistance-conferring apple proteins is one of the most important aims for studying apple resistance mechanisms and promoting the development of disease-resistant apple strains. In order to find proteins which promote resistance to Marssonina coronaria, a deadly pathogen which has been related to premature apple maturation, proteomes from apple leaves inoculated with M. coronaria were characterized at 3 and 6 days post-inoculation by two dimensional electrophoresis (2-DE).
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Anthocyanin contributes more to hydrogen peroxide scavenging than other phenolics in apple peel.
Food Chem
PUBLISHED: 01-22-2014
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The phenolic compounds in apple peel extracts were quantified in the presence of hydrogen peroxide (H2O2) to identify which phenolic compound contributed more to H2O2 scavenging. The results showed that the phenolics extracted from 'Golden Delicious' apple peel had a strong ability for scavenging H2O2. After incubating with H2O2 for 30 min, cyanidin-3-galactoside concentrations in the phenolic extract decreased as H2O2 concentrations increased. In contrast, the concentrations of other phenolic compounds remained unchanged. Exogenous application of H2O2 enhanced the synthesis of phenolics, especially anthocyanin, in 'Golden Delicious' apple peel under sunlight. After incubating the peel extract of H2O2-treated apples in the dark for 30 min, the concentration of cyanidin-3-galactoside significantly decreased to a greater extent than that of other phenolic compounds. Based on these data, anthocyanin is more sensitive to H2O2 and contributes more to H2O2 scavenging than other phenolic compounds.
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Proteomic analysis of 'Zaosu' pear (Pyrus bretschneideri Rehd.) and its early-maturing bud sport.
Plant Sci.
PUBLISHED: 01-21-2014
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Maturation of fruits involves a series of physiological, biochemical, and organoleptic changes that eventually make fleshy fruits attractive, palatable, and nutritional. In order to understand the mature mechanism of the early-maturing bud sport of 'Zaosu' pear, we analyzed the differences of proteome expression between the both pears in different mature stages by the methods of a combination of two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. Seventy-five differential expressed protein spots (p<0.05) were obtained between 'Zaosu' pear and its early-maturing bud sport, but only sixty-eight were demonstratively identified in the database of NCBI and uniprot. The majority of proteins were linked to metabolism, energy, stress response/defense and cell structure. Additionally, our data confirmed an increase of proteins related to cell-wall modification, oxidative stress and pentose phosphate metabolism and a decrease of proteins related to photosynthesis and glycolysis during the development process of both pears, but all these proteins increased or decreased faster in the early-maturing bud sport. This comparative analysis between both pears showed that these proteins were closely associated with maturation and could provide more detailed characteristics of the maturation process of both pears.
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Genome-wide identification of members in the YTH domain-containing RNA-binding protein family in apple and expression analysis of their responsiveness to senescence and abiotic stresses.
Gene
PUBLISHED: 01-11-2014
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YT521-homology (YTH) domain-containing RNA-binding proteins (YTPs) are a small gene family involved in post-transcriptional regulation. We identified 26 putative YTP gene models in the apple genome. Although plant YTPs have been classified into three groups, those in multi-cellular organisms belong only to Groups A and B. The apple genome contains 22 YTP gene models in Group A and four in Group B. Duplication analysis showed that tandem and segmental duplications contributed only partially to an expansion in apple YTP numbers. YTH was the only recognizable domain in apple YTPs; its three-dimensional structure implied possible motifs for RNA-binding. After the assembly of expressed sequence tags (ESTs) and gene-cloning, we were able to identify 14 apple YTPs that were expressed in various tissues, especially senescing leaves. Expression analysis showed that these YTPs also responded to several abiotic stresses. Taken together, our genome-wide evaluation provides new insight for further research on the effects of those stresses.
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The gene family of dehydration responsive element-binding transcription factors in grape (Vitis vinifera): genome-wide identification and analysis, expression profiles, and involvement in abiotic stress resistance.
Mol. Biol. Rep.
PUBLISHED: 01-09-2014
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The dehydration responsive element-binding (DREB) proteins play a critical role in plant development and abiotic stress-mediated gene expression. Therefore, they represent one of the most attractive regulons for breeding programs. However, no comprehensive summary of grapevine DREB family genes is available. During this study, 38 VvDREB members were identified from the entire grapevine genome and its expression sequence tag assembly. These were organized into the same subgroups, A1 through A6, as for Arabidopsis DREBs. The VvDREB genes were distributed in 15 out of 19 chromosomes in grapevine. Multiple sequence alignments were performed and a three-dimensional structure was created to demonstrate sequence conservation. Microarray analysis showed potential regulatory roles for VvDREBs in responses to various abiotic stresses, hormone treatments, berry ripening, exposure to light, and bud development. Cis-acting regulatory elements, such as W-box, MYB-binding site, and light-responsive elements, were the most frequently found in the putative promoter regions. Furthermore, microarray transcriptional profiling of grapevine plants that over-expressed VvDREB23 revealed 248 up-regulated and 229 down-regulated genes, with fold-changes of >1.5 when compared with the empty vector control. Gene ontology classifications showed that different genes function in cellular glucan metabolism, lipid transport, the endomembrane system, cell wall structure, and other important metabolic and developmental processes, as well as in the regulation of molecular functions. Our report provides an overview and constitutes a foundation for further study of this VvDREB gene family. All the microarray data and transcription profiling of transgenic versus empty-vector control transformant grapevines were retrieved from the online resources.
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Photosynthetic performance during leaf expansion in Malus micromalus probed by chlorophyll a fluorescence and modulated 820nm reflection.
J. Photochem. Photobiol. B, Biol.
PUBLISHED: 09-18-2013
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The simultaneous measurements of prompt chlorophyll a fluorescence, delayed chlorophyll a fluorescence and modulated 820nm reflection allow collection and correlation of complementary information for the three domains of the photosynthetic electron transport chain - the PSII electron donor side, electron transport between PSII and PSI, and the PSI electron acceptor side. In this study, we used this approach to investigate photochemical activity during Malus micromalus leaf expansion. The results showed that as leaves expanded, the antenna size per reaction center for the two systems became smaller, and the energetic connectivity of PSII units decreased gradually. Meanwhile, the light trapping efficiency of PSII, electron transfer capacity at the donor side of PSII, exchange capacity of PQs at the QB site and the reoxidation capacity of PQH2 were all increased as leaves expanded. However, the capacity of PQH2 reoxidation increased at a slower rate than the exchange capacity of PQs at the QB site. In general, during leaf development, the photochemical activity of both PSII and PSI increased, although the increase in PSII activity was faster relative to PSI. The results from the three independent signals corroborate each other.
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Delay in leaf senescence of Malus hupehensis by long-term melatonin application is associated with its regulation of metabolic status and protein degradation.
J. Pineal Res.
PUBLISHED: 08-04-2013
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Melatonin has an important anti-aging role in plant physiology. We tested the effects of long-term melatonin exposure on metabolic status and protein degradation during natural leaf senescence in trees of Malus hupehensis Rehd. The 2-month regular supplement of 100 ?m melatonin to the soil once every 6 days altered the metabolic status and delayed protein degradation. For example, leaves from treated plants had significantly higher photosynthetic activity, chlorophyll concentrations, and levels of three photosynthetic end products (sorbitol, sucrose, and starch) when compared with the control. The significant inhibition of hexose (fructose and glucose) accumulation possibly regulated the signaling of MdHXK1, a gene for which expression was also repressed by melatonin during senescence. The plants also exhibited better preservation of their nitrogen, total soluble protein, and Rubisco protein concentrations than the control. The slower process of protein degradation might be a result of melatonin-linked inhibition on the expression of apple autophagy-related genes (ATGs). Our results are the first to provide evidence for this delay in senescence based on the metabolic alteration and protein degradation.
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Isolation and developmental expression analysis of L-myo-inositol-1-phosphate synthase in four Actinidia species.
Plant Physiol. Biochem.
PUBLISHED: 06-06-2013
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Myo-inositol (MI) is an important polyol involved in cellular signal transduction, auxin storage, osmotic regulation, and membrane formation. It also serves as a precursor for the production of pinitol, ascorbic acid, and members of the raffinose family. The first committed step for MI formation is catalyzed by L-myo-inositol-1-phosphate synthase (MIPS). We isolated MIPS cDNA sequences from Actinidia eriantha, Actinidia rufa, and Actinidia arguta and compared them with that of Actinidia deliciosa. Each comprised 1533 bp, encoding 510 amino acids with a predicted molecular weight of 56.5 KDa. The MIPS protein was highly conserved in Actinidia, sharing 98.94% identity among species. The MIPS gene was expressed in the flowers, leaves, petioles, and carpopodia. Similarly high levels of expression were detected in the young fruit of all four species. Overall activity of the enzyme was also maximal in young fruit, indicating that this developmental stage is the key point for MI synthesis in Actinidia. Among the four species, A. arguta had the greatest concentration of MI as well as the highest ratios of MI:sucrose and MI:glucose + fructose. This suggests that conversion to MI from carbohydrates was most efficient in A. arguta during early fruit development.
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Phenylpropanoid metabolites and expression of key genes involved in anthocyanin biosynthesis in the shaded peel of apple fruit in response to sun exposure.
Plant Physiol. Biochem.
PUBLISHED: 04-22-2013
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The shaded peel of Fortune (a red cultivar) and Mutsu (a yellow/green cultivar) apple (Malus domestica Borkh.) was exposed to full sun by turning fruit 180° at about one week before harvest to determine the expression of key genes involved in anthocyanin synthesis in response to sunlight exposure and their relationships with the levels of anthocyanins and other phenolics. For the unturned (control) fruit, the shaded peel had lower expression levels of MdMYB10 (a transcriptional factor in the regulation of anthocyanin biosynthesis) and seven structural genes in anthocyanin synthesis (MdPAL, MdCHS, MdCHI, MdF3H, MdDFR1, MdLDOX, and MdUFGT), and lower levels of anthocyanins and flavonols than the sun-exposed peel in both cultivars. Exposure of the shaded peel to full sun caused marked up-regulation of the expression of MdMYB10 and all seven structural genes, which peaked between 6 h and 30 h after fruit turning, consequently leading to higher levels of anthocyanins, flavonols, and total phenolics than in the shaded peel and even in the sun-exposed peel of control fruit. Interestingly, the levels of flavonols were higher in the shaded peel of turned fruit (the original sun-exposed peel) than in the sun-exposed peel of both control and turned fruit in both cultivars, suggesting that competition for substrates exists in different branches of the phenylpropanoid pathway. These results indicate that sunlight exposure stimulates the expression of MdMYB10 and structural genes in anthocyanin synthesis, thereby elevating the levels of anthocyanins and other phenolic compounds in both red and yellow/green cultivars.
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Light and abiotic stresses regulate the expression of GDP-L-galactose phosphorylase and levels of ascorbic acid in two kiwifruit genotypes via light-responsive and stress-inducible cis-elements in their promoters.
Planta
PUBLISHED: 04-18-2013
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Ascorbic acid (AsA) plays an essential role in plants by protecting cells against oxidative damage. GDP-L-galactose phosphorylase (GGP) is the first committed gene for AsA synthesis. Our research examined AsA levels, regulation of GGP gene expression, and how these are related to abiotic stresses in two species of Actinidia (kiwifruit). When leaves were subjected to continuous darkness or light, ABA or MeJA, heat, or a hypoxic environment, we found some correlation between the relative levels of GGP mRNA and AsA concentrations. In transformed tobacco plants, activity of the GGP promoter was induced by all of these treatments. However, the degree of inducibility in the two kiwifruit species differed among the GGP promoter deletions. We deduced that the G-box motif, a light-responsive element, may have an important function in regulating GGP transcripts under various light conditions in both A. deliciosa and A. eriantha. Other elements such as ABRE, the CGTCA motif, and HSE might also control the promoter activities of GGP in kiwifruit. Altogether, these data suggest that GGP expression in the two kiwifruit species is regulated by light or abiotic stress via the relative cis-elements in their promoters. Furthermore, GGP has a critical role in modulating AsA concentrations in kiwifruit species under abiotic stresses.
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Enhanced salt resistance in apple plants overexpressing a Malus vacuolar Na+/H+ antiporter gene is associated with differences in stomatal behavior and photosynthesis.
Plant Physiol. Biochem.
PUBLISHED: 04-04-2013
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High salinity is a major abiotic factor that limits crop production. The dwarfing apple rootstock M.26 is sensitive to such stress. To obtain an apple that is adaptable to saline soils, we transformed this rootstock with a vacuolar Na(+)/H(+) antiporter, MdNHX1. Differences in salt tolerance between transgenic and wild-type (WT) rootstocks were examined under field conditions. We also compared differences when Naganofuji No. 2 apple was grafted onto these transgenic or WT rootstocks. Plants on the transgenic rootstocks grew well during 60 d of mild stress (100 mM NaCl) while the WT exhibited chlorosis, inhibited growth and even death. Compared with the untreated control, the stomatal density was greater in both non-grafted and grafted WT plants exposed to 200 mM NaCl. In contrast, that density was significantly decreased in leaves from grafted transgenic plants. At 200 mM NaCl, net photosynthesis, stomatal conductance, intercellular CO2 concentration, and chlorophyll contents were markedly reduced in the WT, whereas the declines in those values were only minor in similarly stressed transgenic plants. Therefore, we conclude that overexpressing plants utilize a better protective mechanism for retaining higher photosynthetic capacity. Furthermore, this contrast in tolerance and adaptability to stress is linked to differences in stomatal behavior and photosynthetic rates.
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The role of anthocyanin in photoprotection and its relationship with the xanthophyll cycle and the antioxidant system in apple peel depends on the light conditions.
Physiol Plant
PUBLISHED: 02-06-2013
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The synthesis of anthocyanin, the xanthophyll cycle, the antioxidant system and the production of active oxygen species (AOS) were compared between red and non-red apple cultivars, in response to either long-term sunlight exposure (high light intensity) during fruit development, or to exposure of bagged fruits to lower light intensity late in fruit development. During fruit development of red and non-red apples, the xanthophyll cycle pool size decreased much more in red apple peel late in development. With accumulation of AOS induced by long-term sunlight exposure, enhancement of the antioxidant system was found. However, this change became significantly lower in red apple than non-red apple as fruit developed, which might serve to accelerate the anthocyanin synthesis in red apple peel. When, late in fruit development, bagged fruits were exposed to sunlight, the accumulation of AOS was lower in red apple peel than in non-red peel. This could be due to the higher anthocyanin concentration in the red peels. Meanwhile, compared with that in non-red cultivar, the xanthophyll cycle and the antioxidant system in red apple peel were protected first but then down-regulated by its higher anthocyanin concentration during sunlight exposure. In conclusions, red and non-red apples peel possess different photoprotective mechanisms under high light conditions. The relationship between anthocyanin synthesis and the xanthophyll cycle, and the antioxidant system, depends on the light conditions that fruit undergoes.
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Ultralow-limit gas detection in nano-dumbbell polymer sensor via electrospinning.
Nanoscale
PUBLISHED: 01-31-2013
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Nano-dumbbells via electrospinning: controllable nano-dumbbells are fabricated via electrospinning. The weight block parts and the length of nanowires between them can be adjusted through changing the experimental conditions. This nanostructure enables ultralow-limit gas sensing properties of the resulting polypyrrole-based microsensor.
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Exogenous melatonin improves Malus resistance to Marssonina apple blotch.
J. Pineal Res.
PUBLISHED: 01-28-2013
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We examined whether exogenously applied melatonin could improve resistance to Marssonina apple blotch (Diplocarpon mali) by apple [Malus prunifolia (Willd.) Borkh. cv. Donghongguo]. This serious disease leads to premature defoliation in the main regions of apple production. When plants were pretreated with melatonin, resistance was increased in the leaves. We investigated the potential roles for melatonin in modulating levels of hydrogen peroxide (H2O2), as well the activities of antioxidant enzymes and pathogenesis-related proteins during these plant-pathogen interactions. Pretreatment enabled plants to maintain intracellular H2O2 concentrations at steady-state levels and enhance the activities of plant defence-related enzymes, possibly improving disease resistance. Because melatonin is safe and beneficial to animals and humans, exogenous pretreatment might represent a promising cultivation strategy to protect plants against this pathogen infection.
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Quenching of the electrochemiluminescence of tris(2,2-bipyridine)ruthenium(II)/tri-n-propylamine by pristine carbon nanotube and its application to quantitative detection of DNA.
Anal. Chem.
PUBLISHED: 01-24-2013
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In this study, we describe the quenching of electrochemiluminescence (ECL) of tris(2,2-bipyridine)-ruthenium(II)(Ru(bpy)(3)(2+))/tri-n-propylamine(TPA) at pristine multiwall carbon nanotube (MWNT) modified glassy carbon (GC) electrode. Even though the faradic current of the Ru(bpy)(3)(2+)/TPA system and the oxidation of TPA obtained at pristine MWNT-modified GC electrode is enhanced compared with those at the bare GC electrode, the intensity of ECL produced at MWNT electrode is smaller than that at GC electrode. For testing the possible reason of quenching, a comparison of ECL behavior of Ru(bpy)(3)(2+)/TPA at pristine MWNT and acid-treated, heat-treated, and polyethylene glycol (PEG)-wrapped MWNT-modified GC electrode is studied. The results demonstrate that the oxygen-containing groups at the surface of MWNT and the intrinsic electron properties of MWNT are considered to be the major reason for the suppression of ECL. The comparison also demonstrates that this quenching is related to the distance between MWNT and Ru(bpy)(3)(2+)/TPA. Utilizing this essential quenching mechanism, a new signal-on DNA hybridization assay is proposed on the basis of the MWNT modified electrode, where single-stranded DNA (ssDNA) labeled with Ru(bpy)(3)(2+) derivatives probe (Ru-ssDNA) at the distal end is covalently attached onto the MWNT electrode. ECL signal is quenched where Ru-ssDNA is self-organized on the surface of MWNT electrode; however, the quenched ECL signal returns in case of the presence of complementary ssDNA. The developed approach for sequence-specific DNA detection has good selectivity, sensitivity, and signal-to-background ratio. Therefore, the quenching of the ECL of Ru(bpy)(3)(2+)/TPA system by the pristine MWNT can be an excellent platform for nucleic acid studies and molecular sensing.
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Delayed senescence of apple leaves by exogenous melatonin treatment: toward regulating the ascorbate-glutathione cycle.
J. Pineal Res.
PUBLISHED: 10-12-2011
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The objectives of this study were to test the effects of exogenous melatonin on apple (Malus domestica Borkh. cv. Golden Delicious) leaves and investigate its possible physiological role in delaying leaf senescence. Detached leaves treated with 10?mm melatonin solutions clearly showed a slowing in their process of dark-induced senescence, as evidenced by both biochemical and molecular parameters. Melatonin delayed the normal reduction in chlorophyll content and maximum potential photosystem II efficiency (F(v) /F(m) ). It also suppressed the transcript levels of a key chlorophyll degradation gene, pheide a oxygenase (PAO), and the senescence-associated gene 12 (SAG12). This outcome was thought to be because of the enhanced antioxidant capabilities of melatonin. Indeed, H(2) O(2) accumulation was inhibited by exogenous melatonin, which might have resulted from direct reactive oxygen species scavenging by melatonin and a great enhancement of ascorbate peroxidase (APX; EC 1.11.1.11), which acted on both mRNA and protein activity levels. Melatonin treatment led to the maintenance of higher contents of ascorbic acid (AsA) and glutathione (GSH) but less dehydroascorbate (DHA) and oxidized glutathione (GSSG) compared with the control, possibly through its regulation of the AsA-GSH cycle.
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Influence of drought stress on the cellular ultrastructure and antioxidant system in leaves of drought-tolerant and drought-sensitive apple rootstocks.
Plant Physiol. Biochem.
PUBLISHED: 10-06-2011
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We compared two apple rootstocks -Malus prunifolia and Malus hupehensis - that differ in their tolerance to this abiotic stress. The former is considered drought-tolerant, the latter, sensitive. We monitored changes in their leaf ultrastructure and responses by their antioxidant defense systems. Irrigation was withheld for 12 d from two-year-old potted plants. Compared with the control, this treatment led to considerable ultrastructural alterations in organelles. Plants of M. prunifolia maintained their structural cell integrity longer than did M. hupehensis. M. hupehensis was more vulnerable to drought than was M. prunifolia, resulting in larger increases in the levels of H(2)O(2), O(2)(-), and MDA from the former. Except for catalase (CAT) and monodehydroascorbate reductase (MDHAR), the activities of superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), glutathione reductase (GR), and dehydroascorbate reductase (DHAR) analyzed here were enhanced to a greater extent in M. prunifolia than in M. hupehensis in response to drought. This was also true for levels of ascorbic acid (AsA) and glutathione (GSH). Under well-watered conditions, changes in lipid peroxidation and relevant antioxidant parameters were not significantly different between the two species throughout the experimental period. These results demonstrate that, in order to minimize oxidative damage, both the activities of antioxidant enzymes and antioxidant concentrations are increased in the leaves of M. prunifolia and M. hupehensis in response to water stress. Moreover, plants of M. prunifolia exhibit higher antioxidant capacity and a stronger protective mechanism, such that their cell structural integrity is better maintained during exposure to drought.
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Facile patterning of reduced graphene oxide film into microelectrode array for highly sensitive sensing.
Anal. Chem.
PUBLISHED: 07-27-2011
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In this study, we develop a new technique to fabricate a reduced graphene oxide (rGO)-based microelectrode array (MEA) with low-cost soft lithography. To prepare patterned rGO, a polydimethylsiloxane (PDMS) mold with an array of microwells on its surface is fabricated using soft lithography, and GO is assembled on an indium tin oxide (ITO) electrode with a layer-by-layer method. The rGO pattern is formed by closely contacting the assembled GO film onto the ITO electrode with the PDMS mold filled with hydrazine solution in the microwells to selectively reduce the localized GO into the rGO. The MEA with patterned rGO as the microelectrode is characterized with Kelvin probe force microscopy (KFM), atomic force microscopy (AFM), and cyclic voltammetry (CV) with ferricyanide in aqueous solution as the redox probe. The KFM and AFM results demonstrate that each rGO pattern prepared under the present conditions is 3 ?m in diameter, which is close to that of the PDMS mold we use. The CV results show that the rGO patterned onto the ITO exhibits a sigmoid-shaped voltammogram up to 200 mVs(-1) with a microampere level current response, suggesting that the rGO-based electrode fabricated with soft lithography behalves like a MEA. To demonstrate the potential electroanalytical application of the rGO-based MEA, prussian blue (PB) is electrodeposited onto the rGO-based MEA to form the PB/rGO-based MEA. Electrochemical studies on the formed PB/rGO-based MEA reveal that MEA shows a lower detection limit and a larger current density for the detection of H(2)O(2), as compared with the macroscopic rGO electrode. The method demonstrated here provides a simple and low-cost strategy for the fabrication of graphene-based MEA that are useful for electroanalytical applications.
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Molecular characterization and expression analysis of a glycine-rich RNA-binding protein gene from Malus hupehensis Rehd.
Mol. Biol. Rep.
PUBLISHED: 07-11-2011
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Members of the plant glycine-rich RNA-binding protein (GR-RBP) family play diverse roles in regulating RNA metabolism for various cellular processes. To understand better their function at the molecular level in stress responses, we cloned a GR-RBP gene, MhGR-RBP1, from Malus hupehensis. Its full-length cDNA is 558 bp long, with a 495-bp open reading frame, and it encodes 164 amino acids. The deduced amino acid sequence contains an RNA-recognition motif (RRM) at the amino terminal and a glycine-rich domain at the carboxyl terminal; these are highly homologous with those from other plant species. Multiple alignment and phylogenetic analyses show that the deduced protein is a novel member of the plant GR-RBP family. To characterize this gene, we also applied a model for predicting its homology of protein structure with other species. Both organ-specific and stress-related expression were detected by quantitative real-time PCR and semi-quantitative RT-PCR, indicating that MhGR-RBP1 is expressed abundantly in young leaves but weakly in roots and shoots. Transcript levels in the leaves were increased markedly by drought, hydrogen peroxide (H(2)O(2)), and mechanical wounding, slightly by salt stress. Furthermore, the transcript is initially up- and down-regulated rapidly within 24 h of abscisic acid (ABA) treatment. After 24 h of ABA and jasmonic acid (JA) treatments with different concentrations, the transcript levels of MhGR-RBP1 were significantly repressed. These results suggest that MhGR-RBP1 may be involved in the responses to abiotic stresses, H(2)O(2), ABA, or JA.
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Shading the whole vines during young fruit development decreases ascorbate accumulation in kiwi.
Physiol Plant
PUBLISHED: 07-27-2010
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We investigated how different lighting conditions affected the levels of ascorbate (AsA), sugar contents, the mRNA expression of genes involved in AsA biosynthesis and recycling and enzyme activity in kiwi fruits. Shaded leaves had dramatically less AsA as well as altered transcript levels and enzyme activities. In contrast, fruits that had been covered directly at various developmental stages showed no changes in those parameters. Fruits had significantly less AsA content before 40 days after anthesis (DAA) when whole vines were shaded only from 0 to 40 DAA, whereas transcript levels of most related genes (except those for GDP-L-galactose-1-phosphate phosphorylase and GDP-mannose pyrophosphorylase) followed a parallel trend. When the shading was removed after 40 DAA, values for the ripening fruits returned to those measured for the control. Such a response, however, was not observed when shading treatments were delayed until after 40 DAA. Fruits were also smaller at harvest when vines were shaded at the earliest time point. The present results suggest that lighting conditions can indirectly affect the capacity of biosynthesis and recycling of AsA in young fruits of kiwi, and this regulation might occur via the interaction of signal from leaves and development of fruit.
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Ascorbate biosynthesis during early fruit development is the main reason for its accumulation in kiwi.
PLoS ONE
PUBLISHED: 07-18-2010
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Ascorbic acid (AsA) is a unique antioxidant as well as an enzyme cofactor. Although it has multiple roles in plants, it is unclear how its accumulation is controlled at the expression level, especially in sink tissues. Kiwifruit (Actinidia) is well-known for its high ascorbate content. Our objective was to determine whether AsA accumulates in the fruits primarily through biosynthesis or because it is imported from the foliage.
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Ascorbic acid formation and profiling of genes expressed in its synthesis and recycling in apple leaves of different ages.
Plant Physiol. Biochem.
PUBLISHED: 01-14-2010
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Ascorbic acid (AsA), as a unique antioxidant and enzyme cofactor, has multiple roles in plants. However, there is very limited information on the mechanism of AsA accumulation and controlling in leaves. In this study, we determined AsA accumulation levels, analyzed expression patterns of the genes involved in synthesizing via l-galactose pathway and recycling as well as enzyme activities in apple (Malus domestica Borkh) leaves with different age. AsA content was found to increase with leaf development, reaching the highest level in 20-day-old leaves. This level was maintained in mature leaves until the dropping in senescent leaves. Comparing with young and senescent leaves, mature leaves had higher capability for AsA synthesis with high expression levels and activity of l-galactose dehydrogenase and l-galactono-1,4-lactone dehydrogenase. The mRNA expression of genes involved in AsA synthesis also showed highest abundance in 20-day-old leaves, though GDP-mannose-3,5-epimerase and l-galactose-1-phosphate phosphatase expression reached the highest levels before 20 days old. These results suggest that AsA accumulation in apple leaves mainly occurs during the transition phase from young to mature leaves with high rates of synthesis and recycling, and that l-galactose-1-phosphate phosphatase could play an important role in regulating AsA biosynthesis via the l-galactose pathway.
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Phthalic acid induces oxidative stress and alters the activity of some antioxidant enzymes in roots of Malus prunifolia.
J. Chem. Ecol.
PUBLISHED: 03-11-2009
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Apple replant is a widespread agricultural problem documented in all of the major fruit-growing regions of the world. In order to better understand the phytotoxic mechanisms induced by allelochemicals involved with this problem, Malus prunifolia plants were grown hydroponically to the six-leaf-stage in the presence of phthalic acid (0 or 1 mM) for 5, 10, or 15 days. Apple plants were evaluated for: shoot and root length, fresh and dry weight, malondialdehyde (MDA) content, hydrogen peroxide (H(2)O(2)) content, superoxide radical (O(2) (*-)) generation rate, and antioxidant enzyme activities. Shoot and root lengths and fresh and dry weights of M. prunifolia decreased in plants exposed to phthalic acid. MDA and H(2)O(2) content increased in phthalic acid-treated plants as did the generation rate of O(2) (*-) in M. prunifolia roots. The activities of superoxide dismutase (EC 1.15.1.1), peroxidase (EC 1.11.1.7), catalase (EC 1.11.1.6), ascorbate peroxidase (EC 1.11.1.11), glutathione reductase (EC 1.6.4.2), dehydroascorbate reductase (EC 1.8.5.1), and monodehydroascorbate reductase (EC 1.6.5.4) increased in phthalic acid-stressed roots compared with control roots. These results suggest that activation of the antioxidant system by phthalic acid led to the formation of reactive oxygen species that resulted in cellular damage and the decrease of M. prunifolia growth.
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Influence of light on ascorbate formation and metabolism in apple fruits.
Planta
PUBLISHED: 03-11-2009
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To further understand the regulatory mechanism of light on the formation of ascorbic acid (AsA) in the sink organs of plants, a systematical investigation on AsA levels, activities of two key biosynthsis enzymes and their mRNA expression as well as the recycling was performed in the fruits of apple (Malus domestica Borkh), under different levels of shade. After the whole trees were shaded with the sun-light about 50-55% for 20 days, AsA levels were significantly decreased in fruit peel, flesh and leaves, while mRNA expression levels and activities of L-galactose dehydrogenase (L-GalDH, EC 1.1.1.117) and L-galactono-1,4-lactone dehydrogenase (L-GalLDH, EC 1.3.2.3) as well as activities of recycling enzymes was clearly declined in the leaf and peel but not in the flesh. By shading fruits only for 20 days, AsA levels, relative mRNA levels and activities of L-GalDH and L-GalLDH as well as activities of recycling enzymes all showed obvious decrease in the peel, but not in the flesh. However, their levels in the peel were markedly increased after the full shade was removed and re-exposed these fruits on natural light for 5 days. It is concluded that light affects AsA biosynthesis and recycling in the peel and leaf, but did not in the fresh. Results also suggest that apple fruit is potential to biosynthesize AsA via the L-galactose pathway, and AsA content in the fruits may depend partly on levels of AsA or other photochemistry controlled by light in the leaves.
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Long-term exogenous application of melatonin delays drought-induced leaf senescence in apple.
J. Pineal Res.
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To examine the potential roles of melatonin in drought tolerance, we tested the effects of its long-term exogenous application on Hanfu apple (Malus domestica Borkh.). When 100 ?m melatonin was added to soils under drought conditions, the resultant oxidative stress was eased and leaf senescence was delayed. This molecule significantly reduced chlorophyll degradation and suppressed the up-regulation of senescence-associated gene 12 (SAG12) and pheophorbide a oxygenase (PAO). Such treatment also alleviated the inhibition of photosynthesis brought on by drought stress. We also investigated quenching and the efficiency of Photosystem II (PSII) photochemistry under dark and light conditions and found that melatonin helped to maintain better function of PSII under drought. The addition of melatonin also controlled the burst of hydrogen peroxide, possibly through direct scavenging and by enhancing the activities of antioxidative enzymes and the capacity of the ascorbate-glutathione cycle. Thus, understanding this effect of melatonin on drought tolerance introduces new possibilities to use this compound for agricultural purposes.
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Expression patterns and promoter characteristics of the gene encoding Actinidia deliciosa L-galactose-1-phosphate phosphatase involved in the response to light and abiotic stresses.
Mol. Biol. Rep.
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The gene encoding L-galactose-1-phosphate phosphatase (GPP) plays a central role in ascorbic acid (AsA) biosynthesis in plants. Here, we report AsA contents, GPP expression, and functioning of its promoter in response to light, exogenous stress-signalling hormones, or abiotic stresses in kiwifruit (Actinidia deliciosa). To identify the upstream region of GPP required for promoter activity, we constructed a series of promoter deletion derivatives. Each construct was analyzed by Agrobacterium-mediated transient transformation in tobacco leaves after various treatments. Some correlation was observed between the relative levels of GPP mRNA and AsA contents when kiwi leaves were exposed to varying light conditions, treatment with ABA or SA, wounding, or a hypoxic environment. Analysis of a series of 5 deletions in tobacco leaves indicated that the proximal area 390 bp from the transcription initiation site was needed for establishing both the constitutive and the induced patterns of expression. This promoter was induced by light or one of our abiotic treatments. These results suggest that GPP is regulated by light or abiotic stress and that it plays an important role in controlling AsA contents in kiwifruit.
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Molecular cloning and characterization of a cDNA encoding kiwifruit L-myo-inositol-1-phosphate synthase, a key gene of inositol formation.
Mol. Biol. Rep.
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L-myo-inositol-1-phosphate synthase (MIPS; EC 5.5.1.4) is the key enzyme involved in de novo synthesis of myo-inositol, leading to numerous cellular functions. We isolated an open reading frame of Actinidia deliciosa MIPS (AdMIPS), which is 1,533 bp long and codes for 510 amino acids, with a predicted molecular weight of 56.3 kDa. Sequence analysis revealed its high similarity with MIPS proteins from other organisms. Gene expression and enzyme activity were highest in flower and young fruit. Transcription of AdMIPS was also detected in other tissues. Moderate drought drastically induced expression in the leaves whereas salinity stress induced transcription and enzyme activity in the leaves, phloem, and roots with different degrees. However, a longer period of saline exposure suppressed both expression and enzyme activity in all sampled tissues, indicating that AdMIPS is salt-sensitive.
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Genome-wide identification and expression profiling of dehydrin gene family in Malus domestica.
Mol. Biol. Rep.
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The family of dehydrin genes has important roles in protecting higher plants against abiotic stress, such as drought, salinity and cold. However, knowledge about apple dehydrin gene family is limited. In the present study, we used a bioinformatics approach to identify members of that family in apple (Malus domestica). A total of 12 apple dehydrin genes (MdDHNs) were identified and located on various chromosomes. All putative proteins from those genes contained a typical K domain. Among 12 MdDHNs, nine were cloned and their expression patterns were investigated. Expression profiling indicated that the these nine dehydrin genes display differential expression patterns in various tissues. Moreover, transcript levels of some MdDHNs were up-regulated significantly under drought, low temperature, or ABA treatment, which indicated their important roles during stress adaptation. These results demonstrate that the apple dehydrin gene family may function in tissue development and plant stress responses.
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Primary and secondary metabolism in the sun-exposed peel and the shaded peel of apple fruit.
Physiol Plant
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The metabolism of carbohydrates, organic acids, amino acids and phenolics was compared between the sun-exposed peel and the shaded peel of apple fruit. Contents of sorbitol and glucose were higher in the sun-exposed peel, whereas those of sucrose and fructose were almost the same in the two peel types. This was related to lower sorbitol dehydrogenase activity and higher activities of sorbitol oxidase, neutral invertase and acid invertase in the sun-exposed peel. The lower starch content in the sun-exposed peel was related to lower sucrose synthase activity early in fruit development. Dark respiratory metabolism in the sun-exposed peel was enhanced by the high peel temperature due to high light exposure. Activities of most enzymes in respiratory metabolism were higher in the sun-exposed peel, but the concentrations of most organic acids were relatively stable, except pyruvate and oxaloacetate. Due to the different availability of carbon skeletons from dark respiration in the two peel types, amino acids with higher C/N ratios are accumulated in the sun-exposed peel whereas those with lower C/N ratios are accumulated in the shaded peel. Contents of anthocyanins and flavonols and activities of phenylalanine ammonia-lyase, UDP-galactose:flavonoid 3-O-glucosyltransferase and several other enzymes were higher in the sun-exposed peel than in the shaded peel, indicating the entire phenylpropanoid pathway is upregulated in the sun-exposed peel. Comprehensive analyses of the metabolites and activities of enzymes involved in primary metabolism and secondary metabolism have allowed us to gain a full picture of the metabolic network in the two peel types under natural light exposure.
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Contrasting hypoxia tolerance and adaptation in Malus species is linked to differences in stomatal behavior and photosynthesis.
Physiol Plant
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We examined the potential differences in tolerance to hypoxia by two species of apple rootstocks. Stomatal behavior and photosynthesis were compared between Malus sieversii and Malus hupehensis. Plants were hydroponically grown for 15 days in normoxic or hypoxic nutrient solutions. Those of M. sieversii showed much greater sensitivity, with exposure to hypoxia resulting in higher leaf concentrations of abscisic acid (ABA) that prompted stomatal closure. Compared with the control plants of that species, stomatal density was greater in both new and mature leaves under stress conditions. In contrast, stomatal density was significantly decreased in leaves from M. hupehensis, while stomatal length was unaffected. Under stress, the net photosynthetic rate, stomatal conductance and chlorophyll contents were markedly reduced in M. sieversii. The relatively hypoxia-tolerant genotype M. hupehensis, however, showed only minor changes in net photosynthesis or chlorophyll content, and only a slight decrease in stomatal conductance due to such treatment. Therefore, we conclude that the more tolerant M. hupehensis utilizes a better protective mechanism for retaining higher photosynthetic capacity than does the hypoxia-sensitive M. sieversii. Moreover, this contrast in tolerance and adaptation to stress is linked to differences in their stomatal behavior, photosynthetic capacity and possibly their patterns of native distribution.
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Partitioning of absorbed light energy differed between the sun-exposed side and the shaded side of apple fruits under high light conditions.
Plant Physiol. Biochem.
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Fractions of absorbed light energy consumed via photochemistry and different thermal dissipation processes was quantified and compared between the sun-exposed peel and the shaded peel of apple fruits at different developmental stages. During fruit development, the fraction of absorbed light consumed via photochemistry was no more than 7% in the sun-exposed peel and no more than 5% in the shaded peel under high light conditions. Under high light, the fraction of absorbed light energy consumed via light dependent thermal dissipation was higher whereas that via constitutive thermal dissipation was lower in the sun-exposed peel. The light dependent thermal dissipation in the sun-exposed peel mainly depended on the xanthophyll cycle, and the xanthophyll cycle pool size was significantly larger in the sun-exposed peel than in the shaded peel. The light dependent thermal dissipation in the shaded peel was dependent on both the xanthophyll cycle and the presence of inactivated reaction centers. Under high light conditions, the densities of both Q(A)-reducing reaction centers and Q(B)-reducing reaction centers decreased faster in the shaded peel than in the sun-exposed peel. The thermal dissipation related to photoinhibition increased and then kept unchanged in the sun-exposed peel but decreased in the shaded peel during fruit development. We conclude that under high light intensities, fruit peel looses the excess energy in order of predominance: first by the xanthophyll cycle, then the thermal dissipation related to photoinhibition, next through inactivated reaction centers, and finally by constitutive thermal dissipation.
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Differential expression of ion transporters and aquaporins in leaves may contribute to different salt tolerance in Malus species.
Plant Physiol. Biochem.
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Maintaining ion and water homeostasis in plants is an important defense strategy against salinity stress. Divergence in ion homeostasis between the salt-tolerant Malus hupehensis Rehd. and salt-sensitive Malus prunifolia yingyehaitang was studied to understand their mechanisms for tolerance. Compared with the control on Day 15, plants of those two genotypes under high-salinity treatment had less K(+) in the leaves, stems, and roots. Contents were higher in the roots but lower in the leaves of M. hupehensis while levels in the stems were similar to those from M. prunifolia. For both genotypes, the sodium content increased after salinity treatment in all tissue types. However, the leaves from M. hupehensis had less Na(+) and maintained a lower Na(+)/K(+) ratio. To understand the basis for these differences, we studied the ion transporters and regulation of aquaporin transcripts in the leaves. Transcript levels for both MdHKT1 and MdSOS1 were higher in M. hupehensis, implying that this species had better capacity to exclude sodium so that less Na(+) occurred in the leaves but more in the stems. M. hupehensis also had a greater amount of MdNHX1 transcripts, which could have assisted in sequestering excess Na(+) into the vacuoles and sustaining a better cellular environment. A relatively higher level of aquaporin transcript was also found in M. hupehensis, suggesting that those plants were more capable of maintaining a better leaf water status and diluting excess ions effectively under high-salinity conditions. Therefore, these tested transporters may play important roles in determining how salinity tolerance is conferred in Malus species.
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Superconductivity above 30 K in alkali-metal-doped hydrocarbon.
Sci Rep
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The recent discovery of superconductivity with a transition temperature (T(c)) at 18?K in K(x)picene has extended the possibility of high-T(c) superconductors in organic materials. Previous experience based on similar hydrocarbons, like alkali-metal doped phenanthrene, suggested that even higher transition temperatures might be achieved in alkali-metals or alkali-earth-metals doped such polycyclic-aromatic-hydrocarbons (PAHs), a large family of molecules composed of fused benzene rings. Here we report the discovery of high-T(c) superconductivity at 33?K in K-doped 1,2:8,9-dibenzopentacene (C(30)H(18)). To our best knowledge, it is higher than any T(c) reported previously for an organic superconductor under ambient pressure. This finding provides an indication that superconductivity at much higher temperature may be possible in such PAHs system and is worthy of further exploration.
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Genome-wide analysis and expression profiling of the DREB transcription factor gene family in Malus under abiotic stress.
Mol. Genet. Genomics
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The dehydration responsive element binding (DREB) transcription factor family is one of the most promising regulons for genetic engineering of plant responses to abiotic stresses. However, knowledge about apple DREB genes is limited. In the present study, we found, for the first time, 68 MdDREB genes that could be further classified into six subgroups against the entire genome of apple. All putative proteins from those genes contained a typical APETALA 2 domain and shared similar motifs. The predicted MdDREBs were distributed with different densities over 12 chromosomes, with five tandem duplication sites occurring simultaneously. Both Genevestigator and expressed sequence tags were used for preliminary investigations of expression patterns. Results from quantitative real-time PCR showed that transcript levels of some putative MdDREB genes were up-regulated significantly under various abiotic-stress treatments, which indicated their vital roles during stress adaptation. Identifying these genes and profiling their expression provides useful information and constitutes a foundation for their further, practical utilization in apple through gene-transfer techniques.
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The mitigation effects of exogenous melatonin on salinity-induced stress in Malus hupehensis.
J. Pineal Res.
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As an indoleamine molecule, melatonin mediates many physiological processes in plants. We investigated its role in regulating growth, ion homeostasis, and the response to oxidative stress in Malus hupehensis Rehd. under high-salinity conditions. Stressed plants had reduced growth and a marked decline in their net photosynthetic rates and chlorophyll contents. However, pretreatment with 0.1?m melatonin significantly alleviated this growth inhibition and enabled plants to maintain an improved photosynthetic capacity. The addition of melatonin also lessened the amount of oxidative damage brought on by salinity, perhaps by directly scavenging H(2) O(2) or enhancing the activities of antioxidative enzymes such as ascorbate peroxidase, catalase, and peroxidase. We also investigated whether melatonin might control the expression of ion-channel genes under salinity. Here, MdNHX1 and MdAKT1 were greatly up-regulated in the leaves, which possibly contributed to the maintenance of ion homeostasis and, thus, improved salinity resistance in plants exposed to exogenous melatonin.
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Different effects of light irradiation on the photosynthetic electron transport chain during apple tree leaf dehydration.
Plant Physiol. Biochem.
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Effects of light irradiation on the photosynthetic electron transport chain between P680 and P700 in apple tree leaves was probed with chlorophyll a fluorescence transient and 820 nm transmission measurements during dehydration under different light intensities. The results showed that light accelerated the leaf water-loss rate during dehydration. Leaf dehydration lowered the maximum quantum yield of PSII and the far-red light induced maximal transmission change at 820 nm, but increased the relative variable fluorescence intensity at J-step, especially under increasing irradiation conditions. During leaf dehydration, irradiation lowered the relative variable fluorescence intensity at I-step. At the beginning of leaf dehydration, moderate light accelerated the leaf water-loss rate and then lowered the maximal light-trapping efficiency of P???. Upon further dehydration under moderate light or dehydration under high light, light accelerated the water-loss rate and also directly decreased the maximal light-trapping efficiency of P680. The more significant decrease in the exchange capacity of plastoquinones at the Q(B) site was mainly attributed to the faster water-loss rate under moderate light than in the dark. Under high light, irradiation also directly lowered the capacity. The reoxidation of PQH? in the dehydrated leaves was enhanced by the light irradiation. The rapidly decreased contents of P700 + plastocyanin were mainly attributed to the faster water-loss rate under light conditions in contrast with that in the dark. The different effects of light irradiations on the photosynthetic electron transport chain might be involved in the acclimation of apple tree leaves to dehydration.
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Fabrication of ultra-fine nanostructures using edge transfer printing.
Nanoscale
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The exploration of new methods and techniques for application in diverse fields, such as photonics, microfluidics, biotechnology and flexible electronics is of increasing scientific and technical interest for multiple uses over distance of 10-100 nm. This article discusses edge transfer printing--a series of unconventional methods derived from soft lithography for nanofabrication. It possesses the advantages of easy fabrication, low-cost and great serviceability. In this paper, we show how to produce exposed edges and use various materials for edge transfer printing, while nanoskiving, nanotransfer edge printing and tunable cracking for nanogaps are introduced. Besides this, different functional materials, such as metals, inorganic semiconductors and polymers, as well as localised heating and charge patterning, are described here as unconventional "inks" for printing. Edge transfer printing, which can effectively produce sub-100 nm scale ultra-fine structures, has broad applications, including metallic nanowires as nanoelectrodes, semiconductor nanowires for chemical sensors, heterostructures of organic semiconductors, plasmonic devices and so forth.
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