Purification and characterization of a phytase (myo-inositol-hexakisphosphate phosphohydrolase) accumulated in maize (Zea mays) seedlings during germination.
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ABSTRACT:
SUBMITTER: Laboure AM
PROVIDER: S-EPMC1134897 | biostudies-other | 1993 Oct
REPOSITORIES: biostudies-other
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Project description:The aim of this study was to compare the expression patterns of superoxide dismutase genes (sod2, sod3.4, sod9 and sodB) in seedling leaves of the Zea mays L. Tasty Sweet (susceptible) and Ambrozja (relatively resistant) cultivars infested with one of two hemipteran species, namely monophagous Sitobion avenae F. (grain aphid) or oligophagous Rhopalosiphum padi L. (bird cherry-oat aphid). Secondarily, aphid-elicited alternations in the antioxidative capacity towards DPPH (1,1-diphenyl-2-picrylhydrazyl) radical in insect-stressed plants were evaluated. Comprehensive comparison of expression profiles of the four sod genes showed that both insect species evoked significant upregulation of three genes sod2, sod3.4 and sod9). However, aphid infestation affected non-significant fluctuations in expression of sodB gene in seedlings of both maize genotypes. The highest levels of transcript accumulation occurred at 8 h (sod2 and sod3.4) or 24 h (sod9) post-infestation, and aphid-induced changes in the expression of sod genes were more dramatic in the Ambrozja cultivar than in the Tasty Sweet variety. Furthermore, bird cherry-oat aphid colonization had a more substantial impact on levels of DPPH radical scavenging activity in infested host seedlings than grain aphid colonization. Additionally, Ambrozja plants infested by either hemipteran species showed markedly lower antioxidative capacity compared with attacked Tasty Sweet plants.
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Project description:A full-length cDNA of phyA gene of Aspergillus niger, encoding phytase enzyme, was cloned and expressed in E. coli BL21 cells and assayed for its activity. The phyA cDNA consisted of 1404 bp, which encoded 467 amino acid residues. The phytase activity of purified phytase was 826.33 U/mL. The phyA gene under the control of endosperm-specific promoters was transformed into an Indian maize inbred line, UMI29, using particle bombardment-mediated transformation method to generate transgenic maize plants over-expressing phytase in seeds. PCR and GUS analyses demonstrated the presence of transgenes in T0 transgenic plants and their stable inheritance in the T1 progenies. Three transgenic events expressing detectable level of A. niger phytase were characterized by western blot analysis. Phytase activity of 463.158 U/kg of seed was observed in one of the events, JB-UMI29-Z17/2. The phytase activity of transgenic maize seeds was 5.5- to 7-fold higher than the wild-type UMI29 seeds and, consequently, the seeds had 0.6- to 5-fold higher inorganic phosphorus content.
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Project description:This study analyzed changes in the starch structures and in vitro digestion profiles of a specific maize cultivar, Jike 728 (JK728), in Jilin, China, after 0-5 days of germination. The total starch, amylose, and amylopectin contents decreased significantly during germination. The average molecular weight of the starch compounds also decreased significantly during germination. The proportion of amylopectin with a degree of polymerization (DP) of 13-24 significantly decreased, while the relative abundance of amylopectin with DP values of 6-12, 25-36, and 37-60 significantly increased. The X-ray diffraction (XRD) patterns of all samples were characteristic of A-type starch, and the starch relative crystallinity decreased over time. The proportions of slowly digestible starch and resistant starch decreased significantly, while the proportion of rapidly digestible starch increased significantly during germination. Germination is an easy, inexpensive, and low-carbon processing method. This study indicates that germination is an effective way to control the physical properties and digestibility of starch in maize. The changes observed in the physical properties and digestibility of maize starch after germination provide scientists with a platform to understand starch modification mechanisms that might have potential applications on an industrial scale.
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Project description:BackgroundMaize rough dwarf disease (MRDD) is a severe disease that has been occurring frequently in southern China and many other Asian countries. MRDD is caused by the infection of Rice black streaked dwarf virus (RBSDV) and leads to significant economic losses in maize production. To well understand the destructive effects of RBSDV infection on maize growth, comparative proteomic analyses of maize seedlings under RBSDV infection was performed using an integrated approach involving LC-MS/MS and Tandem Mass Tag (TMT) labeling.ResultsIn total, 7615 maize proteins, 6319 of which were quantified. A total of 116 differentially accumulated proteins (DAPs) were identified, including 35 up- and 81 down-regulated proteins under the RBSDV infection. Enrichment analysis showed that the DAPs were most strongly associated with cyanoamino acid metabolism, protein processing in ER, and ribosome-related pathways. Two sulfur metabolism-related proteins were significantly reduced, indicating that sulfur may participate in the resistance against RBSDV infection. Furthermore, 15 DAPs involved in six metabolic pathways were identified in maize under the RBSDV infection.ConclusionsOur data revealed that the responses of maize to RBSDV infection were controlled by various metabolic pathways.
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Project description:It has long been known that hormones affect the interaction of a phytopathogen with its host plant. The pathogen can cause changes in plant hormone homeostasis directly by affecting biosynthesis or metabolism in the plant or by synthesizing and secreting the hormone itself. We previously demonstrated that pathogenic fungi of the Fusarium species complex are able to produce three major types of hormones: auxins, cytokinins, and gibberellins. In this work, we explore changes in the levels of these hormones in maize and mango plant tissues infected with Fusarium. The ability to produce individual phytohormones varies significantly across Fusarium species and such differences likely impact host specificity inducing the unique responses noted in planta during infection. For example, the production of gibberellins by F. fujikuroi leads to elongated rice stalks and the suppression of gibberellin biosynthesis in plant tissue. Although all Fusarium species are able to synthesize auxin, sometimes by multiple pathways, the ratio of its free form and conjugates in infected tissue is affected more than the total amount produced. The recently characterized unique pathway for cytokinin de novo synthesis in Fusarium appears silenced or non-functional in all studied species during plant infection. Despite this, a large increase in cytokinin levels was detected in F. mangiferae infected plants, caused likely by the up-regulation of plant genes responsible for their biosynthesis. Thus, the accumulation of active cytokinins may contribute to mango malformation of the reproductive organs upon infection of mango trees. Together, our findings provide insight into the complex role fungal and plant derived hormones play in the fungal-plant interactions.
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Project description:BackgroundSugarcane mosaic virus (SCMV) is an important virus pathogen in crop production, causing serious losses in grain and forage yields in susceptible cultivars. Control strategies have been developed, but only marginal successes have been achieved. For the efficient control of this virus, a better understanding of its interactions and associated resistance mechanisms at the molecular level is required.Methodology/principal findingsThe responses of resistant and susceptible genotypes of maize to SCMV and the molecular basis of the resistance were studied using a proteomic approach based on two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS/MS) analysis. Ninety-six protein spots showed statistically significant differences in intensity after SCMV inoculation. The classification of differentially expressed proteins showed that SCMV-responsive proteins were mainly involved in energy and metabolism, stress and defense responses, and photosynthesis. Most of the proteins identified were located in chloroplasts, chloroplast membranes, and the cytoplasm. Analysis of changes in phytohormone levels after virus inoculation suggested that salicylic acid, abscisic acid, jasmonic acid, and azelaic acid may played important roles in the maize response to SCMV infection.Conclusions/significanceAmong these identified proteins, 19 have not been identified previously as virus-responsive proteins, and seven were new and did not have assigned functions. These proteins may be candidate proteins for future investigation, and they may present new biological functions and play important roles in plant-virus interactions. The behavioural patterns of the identified proteins suggest the existence of defense mechanisms operating during the early stages of infection that differed in two genotypes. In addition, there are overlapping and specific phytohormone responses to SCMV infection between resistant and susceptible maize genotypes. This study may provide important insights into the molecular events during plant responses to virus infection.
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Project description:Reduced forms of ascorbate (AsA) and glutathione (GSH) are among the most important non-enzymatic foliar antioxidants in maize (Zea mays L.). The survey was aimed to evaluate impact of bird cherry-oat aphid (Rhopalosiphum padi L.) or grain aphid (Sitobion avenae F.) herbivory on expression of genes related to ascorbate-glutathione (AsA-GSH) cycle in seedlings of six maize varieties (Ambrozja, Nana, Tasty Sweet, Touran, Waza, Złota Karłowa), differing in resistance to the cereal aphids. Relative expression of sixteen maize genes encoding isoenzymes of ascorbate peroxidase (APX1, APX2, APX3, APX4, APX5, APX6, APX7), monodehydroascorbate reductase (MDHAR1, MDHAR2, MDHAR3, MDHAR4), dehydroascorbate reductase (DHAR1, DHAR2, DHAR3) and glutathione reductase (GR1, GR2) was quantified. Furthermore, effect of hemipterans' attack on activity of APX, MDHAR, DHAR and GR enzymes, and the content of reduced and oxidized ascorbate and glutathione in maize plants were assessed. Seedling leaves of more resistant Z. mays varieties responded higher elevations in abundance of target transcripts. In addition, earlier and stronger aphid-triggered changes in activity of APX, MDHAR, DHAR and GR enzymes, and greater modulations in amount of the analyzed antioxidative metabolites were detected in foliar tissues of highly resistant Ambrozja genotype in relation to susceptible Tasty Sweet plants.
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Project description:BackgroundThe present study was designed to investigate the inhibition role of two polyamine biosynthesis inhibitors, i.e., D-arginine (D-Arg) and DL-α-difluoromethylornithine (DFMO), in polyamine biosynthesis under chilling stress in different tissues of two maize inbred lines - Huang C (chilling-tolerance) and Mo17 (chilling-sensitive).ResultsThe results showed that exposure to the lower concentration of polyamine biosynthesis inhibitors improved seedlings growth, such as the root length, root and shoot fresh weight, chlorophyll a (chl a). The effectiveness of 10 µM D-Arg treatments was more prominent than those of 10 µM DFMO. However, the higher concentration of inhibitors suppressed seedlings growth, and the exposure to 100 µM DFMO caused stronger decreases in the photosynthetic pigments, such as chlorophyll a (chl a), chlorophyll b (chl b), total chlorophyll and carotenoids, than the other treatments. Meanwhile, the inhibitor treatments caused the lower content of putrescine (Put) in roots, mesocotyls and coleoptiles in both maize inbred lines as compared with untreated plants. However, the lower concentration (10 µM) of polyamine biosynthetic inhibitors improved the Spd content, except 10 µM D-Arg in root of Huang C, and 10 µM DFMO in coleoptiles of both Mo17 and Huang C. The correlation analysis found that Spd was positively significantly correlated with root length and shoot fresh weight of seedling.ConclusionIt was showed that the Spd played an important role in seedling growth improvement. At the same concentration of polyamine biosynthetic inhibitors, the Put contents in different tissues of the seedlings treated with DFMO were generally lower than those treated with D-Arg, except for Put contents in root of Mo17 with 10 µM treatment. Moreover, the treatments of 100 µM were more prominent than those of 10 µM treatments. Exposure to 100 µM D-Arg and 100 µM DFMO could each decrease the activities of Arginine decarboxylase (ADC), Ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC) in all maize tissues. However, the decrease of the ADC activity was more prominent in 100 µM D-Arg-treated seedlings, while the decrease of SAMDC and ODC activities was prominent in 100 µM DFMO-treated seedlings. Genes involved in polyamine biosynthesis, such as ADC, ODC, SAMDC, and PAO, showed different expression patterns in response to chilling stress and polyamine biosynthesis inhibitors. This study suggested that Put was synthesized via both the ADC and ODC pathways after chilling stress, with the ODC pathway being the major one.
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Project description:Cadmium (Cd) is one of the major heavy metal pollutants in the environment and imposes severe limitations on crop growth and production. Glutathione (GSH) plays an important role in plant Cd tolerance which is able to scavenge stresses-induced reactive oxygen species (ROS) and is involved in the biosynthesis of phytochelatins (PCs). Our previous study revealed that Cd stress affects maize growth, and the GSH treatment could relieve Cd stress in maize seedlings. In this study, we attempted to characterize the metabolomics changes in maize leaves and roots under Cd stress and exogenous GSH conditions. We identified 145 and 133 metabolites in the leaves and roots, respectively. Cd stress decreased the tricarboxylic acid cycle (TCA cycle) metabolism and increased the amino acid contents in the leaves, while it decreased the amino acid contents, increased the TCA cycle metabolism, the sugar contents, and shikimic acid metabolism in the roots. On the other hand, exogenous GSH increased the GSH content, changed the production of metabolites related to antioxidant systems (such as ascorbic acid-related metabolites and flavonoid-related metabolites), and alleviated lipid peroxidation, thereby alleviating the toxic effect of Cd stress on maize. These findings support the idea that GSH alleviates Cd-induced stress in maize and may help to elucidate the mechanism governing Cd-induced stress and the GSH-driven alleviation effect.
| S-EPMC7825527 | biostudies-literature
Project description:Chilling stress greatly inhibited the seed germination, plant growth, development and productivity in this study. The current research aimed to study the effects of different polyamine (PA) inhibitor combinations (Co), e.g., D-arginine (D-Arg), difluoromethylormithine (DFMO), aminoguanidine (Ag) and methylglyoxyl-bis-(guanyhydrazone) (MGBG) at different doses, i.e., 10 µM Co, 100 µM Co, 500 µM Co, 1000 µM Co and 1000 µM Co + 1 mM Spd (Spermidine) in two inbred lines of maize (Zea mays L.), i.e., Mo17 and Huang C, a sensitive and tolerant chilling stress, respectively. The combination treatments of PA inhibitors reduced the biosynthesis of putrescine (Put) in the tissues of both studied inbred lines. Application with 500 µM Co and 1000 µM Co did not result in a significant difference in Put concentrations, except in the coleoptile of Mo17. However, combining Spd to 1000 μM of PA inhibitors enhanced the Put, Spd, spermine (Spm) and total PAs in the roots, coleoptile and mesocotyls. Put and total PAs were increased by 39.7% and 30.54%, respectively, when Spd + 1000 µM Co were applied relative to their controls. Chilling stress and PA inhibitors treatments affected both inbred lines and resulted in differences in the PA contents. Results showed that enzymes involved in the biosynthesis of PAs (ornithine decarboxylase as ODC and S-adenosylmethionine decarboxylase as SAMDC) were significantly downregulated by 1000 µM Co in the tissues of both inbred lines. In contrast, the activity of PAO, a Pas degradation enzyme, was significantly improved by 1000 µM Co under chilling stress. However, Spd + 1000 µM Co significantly improved the activities of ODC and SAMDC and their transcript levels (ODC and SAMDC2). While it significantly downregulated the PAO activity and their relative genes (PAO1, PAO2 and PAO3) under chilling stress. Overall, this study elucidates the specific roles of Spd on the pathway of PA inhibitors and PA biosynthesis metabolism in maize seed development in response to chilling stress. Moreover, the Huang C inbred line was more tolerant than Mo17, which was reflected by higher activities of PA biosynthesis-related enzymes and lower activities of PAs' degradative-related enzymes in Huang C.
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