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Exogenous melatonin confers the chilling tolerance of banana fruit by promoting reactive oxygen species (ROS) scavenging system and inducing the unsaturated fatty acid synthesis. The results showed that melatonin treatment increased the contents of phospholipids, promoted the ROS scavenging enzyme, and restrained the activities of lipoxygenase (LOX), and thus reduced the lipid peroxidation of banana peel. In addition, melatonin treatment increased the flavonoids and proline contents, which was conducive to antioxidant capacity. Interestingly, the enhanced antioxidant capacity is conducive to the stability of unsaturated fatty acids and reduce the enzymatic browning reaction. Moreover, melatonin treatment induced the expression of omega-3/6 fatty acid desaturase and triggered the fatty acid metabolism activity, by which maintained higher contents of unsaturated fatty acid in banana peel. Moreover, melatonin treatment stimulated the accumulation of fatty acids in banana peel, and was involved in alleviating fruit chilling injury.

Objectives To comprehend the epigenetic mechanism of low temperature in delaying senescence of fruit, the changes of DNA methylation patterns of genes related to ethylene biosynthesis and signaling were analyzed in tomato fruit. Results In the present results, the expression level of LeEIN3, SlERF-A1 and LeERT10 decreased, and the expression level of LeCTR1 increased in tomato fruit stored at the low temperature of 11 oC. Meanwhile, the DNA methylation level of CpG island of LeEIN3, SlERF-A1 and LeERT10 increased, and the DNA methylation level of CpG island of LeCTR1 decreased in tomato fruit, respectively. The low temperature suppressed ethylene signaling via changing DNA methylation and gene expression, and delayed senescence of tomato fruit. Conclusions The present study offered valuable information for understanding the role of DNA methylation in senescence of fruit, and provided a foundation for genetic modifying the epigenetic target sites and controlling fruit senescence.

Deoxyribonucleic acid (DNA) methylation plays an important role in fruit ripening and senescence. Here, the role of DNA methylation of the CpG island of SlACS10, LeCTR1, LeEIN3, LeERT10, and SlERF-A1 genes induced by heat treatment (37 °C) in postharvest ripening of tomato fruit was studied. After heat treatment, the firmness and vitamin C content showed higher levels, the loss of aldehydes in volatile components was delayed, and the activities of methylase and demethylase decreased in tomato fruit. Moreover, in heat-treated fruit, significant changes in DNA methylation of SlACS10, LeCTR1, LeEIN3, LeERT10, and SlERF-A1 were induced, the expression of LeERT10 and LeEIN3 was inhibited, the expression of SlERF-A1 was increased, by which ethylene signal transmission might be suppressed and the postharvest ripening of tomato fruit was delayed. The present study provided valuable information for understanding the essential role of DNA methylation in the postharvest ripening of tomato fruit.

The effect of melatonin on chilling injury of banana fruit was investigated. Results showed that melatonin treatment delayed chilling injury and alleviated peel browning. Melatonin treatment increased the contents of phospholipids and unsaturated fatty acid, induced the reactive oxygen species (ROS) scavenging enzyme activities, and reduced the contents of H2O2, O2– of banana fruit. It also induced the miR528 expression, resulting in the down expression of the potential target genes of MaPPO1, MaPPO2 and MaPPO3, and inhibited the PPO activities. Moreover, the phenolic compounds in melatonin treated banana fruit peel were quantitatively analyzed by liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The results indicated that melatonin treatment changed the contents of 63 polyphenolics in banana fruit, and by which involved in delaying chilling injury and alleviating peel browning. In conclusion, melatonin treatment is a potential technique in alleviating chilling injury of banana fruit under low temperature storage.

The miRNAs and their targets involved in senescence of strawberry fruit (Fragaria ananassa L. cv. Zhangji) were analyzed in the present study. In the previous work, three members of miR164 family, mdmmiR164d_ 1ss21AC, mdm-miR164e and mdm-miR164f_1ss21TA, and three of their targets, NAC domain transcriptional regulator superfamily protein, NAC domain containing protein 38 and NAC domain containing protein 87 had been identified by high-throughput sequencing and degradome analysis. In the process of fruit senescence from 0 to 48 h at 4°C storage, the relative levels of mdm-miR164e and mdmmiR164d_1ss21AC expression were significantly increased resulting in decreased expression of NAC genes, and delayed senescence of strawberry fruits. These results suggested that miR164 was involved in strawberry fruit senescence by negatively mediating the expression of NAC transcription factors.

The effect of 1-methylcyclopropene (1-MCP) on inhibiting postharvest blue mold of apple fruit caused by Penicillium expansum and suppressing the growth of P. expansum in vitro was investigated. The treatment of 5 μL L −1 1-MCP significantly decreased disease severity of apple fruit caused by P. expansum and suppressed the mycelial growth and spore germination of P. expansum . After treatment with 1-MCP, lower integrity of plasma membrane in the spores of P. expansum was detected, higher levels of reactive oxygen species (ROS) in the spores and malondialdehyde (MDA) in the mycelium was observed, indicating that 1-MCP treatment enhanced oxidative damage to P. expansum and destroyed the integrity of plasma membrane of spores. This study for the first time demonstrated that 1-MCP suppressed postharvest blue mold of apple fruit by inhibiting the growth of P. expansum .

Anthracnose caused by Colletotrichum gloeosporioides is one of the most important postharvest diseases in mango fruit, often causing huge economic losses. In this study, the effect of 1-methylcyclopropene (1-MCP) against anthracnose in postharvest mango fruit and the mechanisms involved were investigated. 1-MCP induced reactive oxygen species (ROS) generation, damaged the mitochondria and destroyed the integrity of plasma membrane of spores of C. gloeosporioides, significantly suppressing spore germination and mycelial growth of C. gloeosporioides. 1-MCP also decreased the decay incidence and lesion expansion of mango fruit caused by C. gloeosporioides. For the first time this study demonstrated that 1-MCP suppressed anthracnose of postharvest mango fruit by directly inhibiting spore germination and mycelial growth of C. gloeosporioides, thus providing a promising strategy for disease control.

To understand the microRNAs (miRNAs)-mediated mechanism of low temperature delaying non-climacteric strawberry fruit senescence during postharvest period, two small RNA (sRNA) libraries from strawberry fruit stored at low temperature for 24 and 48 h were constructed. A total of 88 known and 1224 novel potential candidate miRNAs were obtained and analyzed. Compared with the expression of miRNAs in strawberry fruit stored for 0 h, 108 miRNAs were up-regulated and 113 were down-regulated in fruit stored at 24 h, and 139 miRNAs were up-regulated and 114 were down-regulated in fruit stored at 48 h. In the process of fruit storage under low temperature, PC-5p-176409_20 repressed abscisic acid (ABA) signaling transduction via the PYR1/PYL1-PP2C-SnRK2 network, miR167 reduced the jasmonic acid (JA) biosynthesis by targeting auxin response factor 8(ARF8), and by which they were involved in delaying fruit senescence. MiR164, miR172, PC-5p-67794_53 and PC-5p-1004_3092 up- or down-regulated the expression of their target genes, NAC transcription factors, APETALA2.7 (AP2.7) transcription factor, alpha/beta-hydrolases superfamily protein and glycosyl hydrolase 9B1, respectively, and also involved in delaying fruit senescence under low temperature. These results give valuable information for understanding the role of miRNA in mediating the fruit senescence at low temperature.

One novel transcription factor gene DnWRKY29 was isolated from Dendrobium officinale, and its functions were identified by its overexpression in tobacco. Compared to the wild type (WT), the DnWRKY29 transgenic tobacco seeds and seedlings showed more sensitivity to salt and drought stresses. The seed germination rate, seedling root length, and fresh weight of transgenic tobacco were lower than in WT. The activities of peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) in transgenic tobacco were decreased, and the content of malondialdehyde (MDA) in transgenic tobacco was increased. Moreover, the expression of DnWRKY29 was up-regulated in Dendrobium seedlings under salt and drought stresses, which was consistent with the function identified by its overexpression in tobacco, indicating that the up-regulation or overexpression of DnWRKY29 impaired plant tolerance to salt and drought stresses. These results provided a new insight to the low tolerance of Dendrobium seedlings to environmental change.

Dendrobium seedlings showed low survival rate when they were transferred from in vitro conditions to greenhouse or field environment. One of the major reasons is their low tolerance to environmental changes. WRKY transcription factors are one of the largest families of transcriptional regulators in plants. They are involved in various biotic and abiotic stress responses. One DnWRKY11 gene was isolated from Dendrobium nobile. To explore the function of DnWRKY11 in Dendrobium defense responses to abiotic stress, it was overexpressed in tobacco. Under salt and drought stresses, the DnWRKY11 transgenic tobacco showed higher germination rate, longer root length, higher fresh weight, higher activities of catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), and lower content of malonidialdehyde (MDA) than the wild type. These results proved the important roles of DnWRKY11 in plant response to drought and salt stresses, and provided a potential gene for improving environmental stress tolerance of Dendrobium seedlings.

Proton (H+) and aluminum (Al3+) toxicities are major factors limiting crop production on acid soils. To study whether salicylic acid (SA) is functional in alleviating protein damage caused by H+ and Al3+ toxicities, an investigation of the antioxidant defense response regulated by SA was carried out on barley (Hordeum vulgare L.) seedlings under H+, Al3+, and combined stresses. It was found that the relative root elongation of seedlings, which grew in the solutions supplemented with SA, was significantly higher than that of seedlings without SA treatment after 24-h treatments with H+, Al3+, and combined stresses. The lesser amount of carbonylated proteins with molecular weights ranging from 14.4 to 97 kD, was accumulated in seedlings treated with SA than that in the seedlings without SA treatment. The higher activities of antioxidant enzymes and lesser content of MDA were observed in seedlings treated with SA compared with the seedlings without SA treatment. Moreover, the nitroblue tetrazolium staining of roots showed that ROS accumulation was decreased by SA treatments. This study suggested that SA could alleviate cell damage caused by H+ and Al3+ toxicities on acid soils by both activating antioxidant defense responses and reducing the contents of carbonylated proteins caused by ROS in barley seedlings.

The genome of Arabidopsis thaliana contains seven Hsp90 family genes. Three organellar and two cytosolic AtHsp90 isoforms were characterized by functionally expressing them in a temperature-sensitive Hsp90 mutant and a conditional Hsp90-null mutant of Saccharomyces cerevisiae. The cytosolic AtHsp90-1 and AtHsp90-2 showed function similar to that of yeast in chaperoning roles; they could support the growth of yeast mutants at both permissive and non-permissive temperature. Neither the full-length nor mature forms of chloroplast-located AtHsp90-5, mitochondria-located AtHsp90-6 and endoplasmic reticulum (ER)-located AtHsp90-7 could complement the yeast Hsp90 proteins. The cytosolic AtHsp90s could stabilize the biomembrane of the temperature-sensitive Hsp90 mutant strains under stress conditions, while the organellar AtHsp90s could not protect the biomembrane of the temperature-sensitive Hsp90 mutant strains. Yeast two-hybrid results showed that either pre-protein or mature forms of organellar AtHsp90s could interact with cofactors cpHsp70, Hsp70, Hsp70t-2, Cyp40, p23 and a substrate protein of NOS, while cytosolic AtHsp90s could not interact with them. These results suggest that organellar and cytosolic AtHsp90s possibly work through different molecular mechanisms in forming chaperone complexes and performing their functional roles.

BACKGROUND: Proton (H+) and aluminium (Al3+) toxicities are major factors limiting crop production on acid soils, while γ-aminobutyric acid (GABA) is a non-protein amino acid involved in various stress tolerances in plants. In this study, to determine whether exogenous GABA is functional in alleviating oxidative stress induced by H+ and Al3+ toxicities, the antioxidant defence response regulated by GABA was investigated in barley (Hordeum vulgare L.). RESULTS: After 24 h treatments of seedlings under H+, Al3+ and combined stresses with and without GABA, morphological and biochemical assays were conducted. It was observed that the inhibition of seedling root elongation caused by Al3+ and H+ toxicities was significantly mitigated by GABA. The amount of carbonylated proteins with molecular masses of 14.4–97 kDa was decreased. The activities of antioxidant enzymes were enhanced, the content of malondialdehyde was reduced and the accumulation of reactive oxygen species (ROS), as shown by staining roots with nitroblue tetrazolium, declined in GABA-treated seedlings. CONCLUSION: GABA can alleviate oxidative damage caused by H+ and Al3+ toxicities in barley seedlings by activating antioxidant defence responses and reducing the elevated levels of carbonylated proteins caused by ROS. Copyright © 2010 Society of Chemical Industry

The PROMOTION OF CELL SURVIVAL 1 (PCS1) gene, encoding an aspartic protease, has an important role in determining the fate of cells in embryonic development and reproduction processes in Arabidopsis. To explore the potential function of the PCS1 gene in generating reproductive sterility, we placed the PCS1 gene under the control of an 1,869-bp nucleotide sequence from the 3′ end of the second intron (AG-I) of Arabidopsis AGAMOUS and CaMV 35S (–60) minimal promoter [AG-I-35S (–60)::PCS1], and introduced it into tobacco. RT–PCR results demonstrated that the PCS1 gene driven by AG-I-35S (–60) chimeric promoter was expressed only in anthers and carpels in the reproductive tissues of transgenic tobacco. Compared to wild-type plants, all AG-I-35S (–60) and AG-I-35S (–60)::PCS1 transgenic lines showed a normal phenotype throughout the vegetative growth phase. However, during the reproductive stage, most AG-I-35S (–60)::PCS1 transgenic plant anthers displayed delayed dehiscence, failed dehiscence, petalody and hypoplasia, and the pollen grains had different shapes and sizes with a distorted, shrunken, or collapsed morphology. Moreover, three transgenic lines, PCS1-1, PCS1-3 and PCS1-4, showed higher sterility than wild-type and AG-I-35S (–60) transgenic plants, respectively. These results showed that the construct of AG-I-35S (–60)::PCS1 was partially effective at preventing seed set and provided a novel sterility strategy.

Three AtHSP90 isoforms, cytosol-localized AtHSP90.2, chloroplast-localized AtHSP90.5, and endoplasmic reticulum (ER)-localized AtHSP90.7 genes, were constitutively overexpressed in Arabidopsis thaliana to study their functional mechanisms under oxidative stress. Overexpression of AtHSP90 genes reduced germination of transgenic seeds under oxidative stress. When exposed to 10 mM H2O2, AtHSP90 transgenic seedlings displayed lower activities of superoxide dismutase, catalase, and peroxidase; higher content of malondialdehyde; and higher levels of protein damage than detected in the wild type. This indicated that overexpression of AtHSP90.2, AtHSP90.5, and AtHSP90.7 in Arabidopsis impaired plant tolerance to oxidative stress. Moreover, overexpression of chloroplast- and ER-localized AtHSP90 resulted in lower resistance to oxidative stress than that of cytosolic AtHSP90. This suggested that HSP90.2, HSP90.5, and HSP90.7 localized in different cellular compartments were involved in different functional mechanisms during oxidative stress.

Three AtHsp90 isoforms, cytosolic AtHsp90.2, chloroplast-located AtHsp90.5, and endoplasmic reticulum (ER)-located AtHsp90.7, were characterized by constitutive overexpressing their genes in Arabidopsis thaliana. Both types of the transgenic plants overexpressing cytosolic and organellar AtHsp90s showed reduced tolerance to salt and drought stresses with lower germination rates and fresh weights, but improved tolerance to high concentration of Ca(2+) comparing with the wild type plants. Transcriptional analysis of ABA-responsive genes, RD29A, RD22 and KIN2 under salt and drought stresses, indicated that the induction expression of these genes was delayed by constitutive overexpression of cytosolic AtHsp90.2, but was hardly affected by that of organellar AtHsp90.5 and AtHsp90.7. These results implied that Arabidopsis different cellular compartments-located Hsp90s in Arabidopsis might be involved in abiotic stresses by different functional mechanisms, probably through ABA-dependent or Ca(2+) pathways, and proper homeostasis of Hsp90 was critical for cellular stress response and/or tolerance in plants.

Aims Dendrobium officinale is an important traditional Chinese medicinal herb. Its seedlings generally show low survival and growth when transferred from in vitro tissue culture to a greenhouse or field environment. In this study, the effect of Mycena dendrobii on the survival and growth of D. officinale tissue culture seedlings and the mechanisms involved was explored. Methods and results Mycena dendrobii were applied underneath the roots of D. officinale tissue culture seedlings. The seedling survival and growth were analysed. The root proteins induced by M. dendrobii were identified using two-dimensional (2-D) electrophoresis and matrix-assisted laser desorption/ionization time-of-flight MS (MALDI-TOF-MS). Mycena dendrobii treatment significantly enhanced survival and growth of D. officinale seedlings. Forty-one proteins induced by M. dendrobii were identified. Among them, 10 were involved in defence and stress response, two were involved in the formation of root or mycorrhizae, and three were related to the biosynthesis of bioactive constituents. Conclusions These results suggest that enhancing stress tolerance and promoting new root formation induced by M. dendrobii may improve the survival and growth of D. officinale tissue culture seedlings. Significance and impact of the study This study provides a foundation for future use of M. dendrobii in the large-scale cultivation of Dendrobiums.

Salvia miltiorrhiza is one of the most popular traditional medicinal herbs in Asian nations. Its dried root contains a number of tanshinones, protocatechuic aldehyde, salvianolic acid B and rosmarinic, and is used for the treatment of various diseases. The finding of microRNAs (miRNAs) and their target genes will help understand their biological role on the biosynthesis of tanshinones in S. miltiorrhiza. In the present study, a total of 452 known miRNAs corresponding to 589 precursor miRNAs (pre-miRNAs), and 40 novel miRNAs corresponding to 24 pre-miRNAs were identified in different tissues of S. miltiorrhiza by high-throughput sequencing, respectively. Among them, 62 miRNAs express only in root, 95 miRNAs express only in stem, 19 miRNAs express only in leaf, and 71 miRNAs express only in flower, respectively. By the degradome analysis, 69 targets potentially cleaved by 25 miRNAs were identified. Among them, acetyl-CoA C-acetyltransferase was cleaved by miR5072, and involved in the biosynthesis of tanshinones. This study provided valuable information for understanding the tissue-specific expression patterns of miRNAs in S. miltiorrhiza, and offered a foundation for future studies of the miRNA-mediated biosynthesis of tanshinones.

The functions of cytosolic heat shock protein AtHsp90.3 in response to heavy metal stress were characterized by using expression of AtHsp90.3 gene in yeast and Arabidopsis thaliana. AtHsp90.3 supported the Saccharomyces cerevisiae Hsp90 knockout strain R0005 growth and maintaining cells membrane integrity under cadmium and arsenic stresses, which was compatible with the components of ScHsc82 machinery. However, constitutive overexpression of AtHsp90.3 in Arabidopsis impaired plant tolerance to Cd stress with lower germination rate and shorter root length, decreased contents of phytochelatins (PCs) and glutathione (GSH), inhibited activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), and increased content of malondialdehyde (MDA). These results suggested that proper homeostasis of Hsp90 was critical for cellular response and/or tolerance to heavy metal stress in plants.

In non-climacteric fruits, the respiratory increase is absent and no phytohormone is appearing to be critical for their ripening process. They must remain on the parent plant to enable full ripening and be picked at or near the fully ripe stage to obtain the best eating quality. However, huge losses often occur for their quick post-harvest senescence. To understanding the complex mechanism of non-climacteric fruits post-harvest senescence, we constructed two small RNA libraries and one degradome from strawberry fruit stored at 20°C for 0 and 24 h. A total of 88 known and 1224 new candidate miRNAs, and 103 targets cleaved by 19 known miRNAs families and 55 new candidatemiRNAs were obtained. These targets were associated with development, metabolism, defense response, signaling transduction and transcriptional regulation. Among them, 14 targets, including NAC transcription factor, Auxin response factors (ARF) and Myb transcription factors, cleaved by 6 known miRNA families and 6 predicted candidates, were found to be involved in regulating fruit senescence. The present study provided valuable information for understanding the quick senescence of strawberry fruit, and offered a foundation for studying the miRNA-mediated senescence of non-climacteric fruits.

Rice seed size is an important agronomic trait in determining the yield potential, and four seed size related genes (GS3, GW2, qSW5/GW5 and GIF1) have been cloned in rice so far. However, the relationship among these four genes is still unclear, which will impede the process of gene pyramiding breeding program to some extent. To shade light on the relationship of above four genes, gene expression analysis was performed with GS3-RNAi, GW2-RNAi lines and CSSL of qSW5 at the transcriptional level. The results clearly showed that qSW5 and GW2 positively regulate the expression of GS3. Meanwhile, qSW5 can be down-regulated by repression of GW2 transcription. Additionally, GIF1 expression was found to be positively regulated by qSW5 but negatively by GW2 and GS3. Moreover, the allelic effects of qSW5 and GS3 were detailedly characterized based on a natural population consisting of 180 rice cultivars. It was indicated that mutual interactions exist between the two genes, in which, qSW5 affecting seed length is masked by GS3 alleles, and GS3 affecting seed width is masked by qSW5 alleles. These findings provide more insights into the molecular mechanisms underlying seed size development in rice and are likely to be useful for improving rice grain yield.

A 1681 bp PsPR10 promoter was isolated from Pinus strobus and a series of 5′-deletions were fused to the β-glucuronidase (GUS) reporter gene and introduced into tobacco. GUS activity in P796 (−796 to +69) construct transgenic plant roots was similar with that of P1681 and higher than those of the P513 (−513 to +69) and P323 (−323 to +69) transgenic plants. Moreover, the abiotic stresses of NaCl, PEG 6000 and mannitol, and salicylic acid (SA), abscisic acid (ABA) and jasmonic acid (JA) induced higher GUS activity in the roots of P796 transgenic tobacco. This study provides a potential inducible root-specific promoter for transgenic plants.

Soil salinity is a major abiotic stress that limits agriculture productivity worldwide. Salicornia europaea is a succulent annual euhalophyte and one of the most salt tolerant plant species. The elucidation of its salt tolerance mechanism is of significance for generating salt-tolerant crops. In this study, we provided high resolution of proteome reference maps of S. europaea shoot and obtained evidence on the salt tolerance mechanism by analyzing the proteomic responses of this plant to high salinity. Our results demonstrated significant variations existed in 196 out of 1880 protein spots detected on CBB stained 2-DE gels. Of these, 111 proteins were identified by mass spectrometry. Among them, the majority was energy production and conversion related proteins, followed by photosynthesis and carbohydrate metabolism associated enzymes. Analysis of protein expression patters revealed that energy production and ion homeostasis associated proteins played important roles for this plant salt tolerance ability. Hierarchical clustering results revealed many proteins were involved in S. europaea salt tolerance mechanism as a dynamic network. Finally, based on our proteomic results, we brought forward a possible schematic representation of mechanism associated with the systematic salt tolerance phenotype in S. europaea.