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Journal ArticleDOI

Isolation and Characterization of a myo-inositol-1-phosphate Synthase Gene from Yellow Passion Fruit (Passiflora edulis f. flavicarpa) Expressed During Seed Development and Environmental Stress

01 Feb 2007-Annals of Botany (Oxford University Press)-Vol. 99, Iss: 2, pp 285-292
TL;DR: Experimental data suggest that PeMIPS1 transcription plays an important role in the establishment of developmental programmes and during the response of plants to environmental changes, suggesting that it is important for environmental stress response.
About: This article is published in Annals of Botany.The article was published on 2007-02-01 and is currently open access. It has received 66 citations till now. The article focuses on the topics: Gene expression & Complementary DNA.
Citations
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Journal ArticleDOI
TL;DR: This study demonstrates for the first time the in vivo interference phenomenon in the pathogenic fungus Fusarium verticillioides, in which expression of an individual fungal transgene was specifically abolished by inoculating mycelial cells in transgenic tobacco plants engineered to express siRNAs from a dsRNA corresponding to the particular transgenes.
Abstract: Self-complementary RNA transcripts form a double-stranded RNA (dsRNA) that triggers a sequence-specific mRNA degradation, in a process known as RNA interference (RNAi), leading to gene silencing. In vascular plants, RNAi molecules trafficking occur between cells and systemically throughout the plant. RNAi signals can spread systemically throughout a plant, even across graft junctions from transgenic to non-transgenic stocks. There is also a great interest in applying RNAi to pathogenic fungi. Specific inhibition of gene expression by RNAi has been shown to be suitable for a multitude of phytopathogenic filamentous fungi. However, double-stranded (ds)RNA/small interfering (si)RNA silencing effect has not been observed in vivo. This study demonstrates for the first time the in vivo interference phenomenon in the pathogenic fungus Fusarium verticillioides, in which expression of an individual fungal transgene was specifically abolished by inoculating mycelial cells in transgenic tobacco plants engineered to express siRNAs from a dsRNA corresponding to the particular transgene. The results provide a powerful tool for further studies on molecular plant-microbe and symbiotic interactions. From a biotechnological perspective, silencing of fungal genes by generating siRNAs in the host provides a novel strategy for the development of broad fungi-resistance strategies in plants and other organisms.

155 citations


Cites methods from "Isolation and Characterization of a..."

  • ...PCRs were carried out as described [ 62 ], except that 20 ng of cDNA was used as a template, in reactions with 32 cycles of amplification....

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Journal ArticleDOI
TL;DR: In this article, the authors found that the expression of the sweet potato IbMIPS1 gene was induced by NaCl, polyethylene glycol (PEG), abscisic acid (ABA), and stem nematodes.
Abstract: Summary Myo-inositol-1-phosphate synthase (MIPS) is a key rate limiting enzyme in myo-inositol biosynthesis. The MIPS gene has been shown to improve tolerance to abiotic stresses in several plant species. However, its role in resistance to biotic stresses has not been reported. In this study, we found that expression of the sweet potato IbMIPS1 gene was induced by NaCl, polyethylene glycol (PEG), abscisic acid (ABA) and stem nematodes. Its overexpression significantly enhanced stem nematode resistance as well as salt and drought tolerance in transgenic sweet potato under field conditions. Transcriptome and real-time quantitative PCR analyses showed that overexpression of IbMIPS1 up-regulated the genes involved in inositol biosynthesis, phosphatidylinositol (PI) and ABA signalling pathways, stress responses, photosynthesis and ROS-scavenging system under salt, drought and stem nematode stresses. Inositol, inositol-1,4,5-trisphosphate (IP3), phosphatidic acid (PA), Ca2+, ABA, K+, proline and trehalose content was significantly increased, whereas malonaldehyde (MDA), Na+ and H2O2 content was significantly decreased in the transgenic plants under salt and drought stresses. After stem nematode infection, the significant increase of inositol, IP3, PA, Ca2+, ABA, callose and lignin content and significant reduction of MDA content were found, and a rapid increase of H2O2 levels was observed, peaked at 1 to 2 days and thereafter declined in the transgenic plants. This study indicates that the IbMIPS1 gene has the potential to be used to improve the resistance to biotic and abiotic stresses in plants.

152 citations


Cites background from "Isolation and Characterization of a..."

  • ...Myo-inositol-1-phosphate synthase is a key rate limiting enzyme of inositol biosynthesis (Abreu and Arag~ao, 2007)....

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  • ...4) is a key rate limiting enzyme of myo-inositol biosynthesis which catalyses the reaction from glucose-6-phosphate (G-6-P) to myo-inositol-1-phosphate (Ins1P), which is subsequently dephosphorylated by myo-inositol monophosphatase (MIPP) to form free inositol (Abreu and Arag~ao, 2007)....

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Journal ArticleDOI
TL;DR: A number of novel proteins were differentially expressed or appeared only in the PEG-fractionated protein samples, indicating that PEG fractionation system can be used as a versatile protein fractionation technique in proteomic analysis to identify novel or low-abundant proteins from all kinds of plant species.
Abstract: A comparative proteomic approach has been adopted in combination with physiological and biochemical analysis of tomato leaves responding to waterlogging stress. Waterlogging resulted in increases of relative ion leakage, lipid peroxidation and in vivo H2O2 content, whereas the chlorophyll content was decreased. Histocytochemical investigations with 3,3'-diaminobenzidine to localize H2O2 and Evans blue to detect dead cells suggested that oxidative stress has a significant role to leaf senescence. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), the most abundant leaf protein, was successfully reduced from the samples by a fractionation method based on 15% polyethylene glycol (PEG). Elimination of Rubisco was further confirmed by Western blot analysis. To elucidate the temporal changes of the protein patterns in tomato leaves, the total soluble and the PEG-fractionated proteins were separated by two-dimensional electrophoresis (2-DE) and visualized by Coomassie Brilliant Blue staining. A total of 52 protein spots were differentially expressed, wherein 33 spots were identified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry or electrospray ionization tandem mass spectrometry (ESI-MS/MS) analysis. The identified proteins are involved in several processes, i.e. photosynthesis, disease resistance, stress and defense mechanisms, energy and metabolism and protein biosynthesis. Results from 2-DE analysis, combined with immunoblotting clearly showed that the fragments of Rubisco large subunit were significantly degraded. This could result from a higher production of reactive oxygen species in leaves under waterlogging stress. Furthermore, four differentially accumulated proteins were analyzed at the mRNA level, confirming the differential gene expression levels and revealing that transcription levels are not always concomitant to the translation level. A number of novel proteins were differentially expressed or appeared only in the PEG-fractionated protein samples, indicating that PEG fractionation system can be used as a versatile protein fractionation technique in proteomic analysis to identify novel or low-abundant proteins from all kinds of plant species.

121 citations


Cites background from "Isolation and Characterization of a..."

  • ...MIPS catalyses the conversion of D-glucose 6-phosphate to 1-myo-inositol-1phosphate, the first and rate-limiting step in the biosynthesis of all inositol containing compounds (Abreu and Aragao 2007)....

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Journal ArticleDOI
TL;DR: Overexpression of MfMIPS1 in tobacco increased the MIPS activity and levels of myo-inositol, galactinol and raffinose, resulting in enhanced resistance to chilling, drought and salt stresses in transgenic tobacco plants.
Abstract: myo-Inositol phosphate synthase (MIPS) is the key enzyme of myo-inositol synthesis, which is a central molecule required for cell metabolism and plant growth as a precursor to a large variety of compounds. A full-length fragment of MfMIPS1 cDNA was cloned from Medicago falcata that is more cold-tolerant than Medicago sativa. While MfMIPS1 transcript was induced in response to cold, dehydration and salt stress, MIPS transcript and myo-inositol were maintained longer and at a higher level in M. falcata than in M. sativa during cold acclimation at 5 °C. MfMIPS1 transcript was induced by hydrogen peroxide (H(2) O(2)) and nitric oxide (NO), but was not responsive to abscisic acid (ABA). Pharmacological experiments revealed that H(2) O(2) and NO are involved in the regulation of MfMIPS1 expression by cold and dehydration, but not by salt. Overexpression of MfMIPS1 in tobacco increased the MIPS activity and levels of myo-inositol, galactinol and raffinose, resulting in enhanced resistance to chilling, drought and salt stresses in transgenic tobacco plants. It is suggested that MfMIPS1 is induced by diverse environmental factors and confers resistance to various abiotic stresses.

111 citations

Journal ArticleDOI
TL;DR: This study compares not only genes that are up- and down-regulated in a drought-tolerant genotype under terminal drought stress and a drought susceptible genotype but also between the bulks of the selected RILs exhibiting extreme phenotypes to provide a better insight into the selection of candidate genes associated with drought tolerance.
Abstract: Chickpea (Cicer arietinum L.) is an important grain-legume crop that is mainly grown in rainfed areas, where terminal drought is a major constraint to its productivity. We generated expressed sequence tags (ESTs) by suppression subtraction hybridization (SSH) to identify differentially expressed genes in drought-tolerant and -susceptible genotypes in chickpea. EST libraries were generated by SSH from root and shoot tissues of IC4958 (drought tolerant) and ICC 1882 (drought resistant) exposed to terminal drought conditions by the dry down method. SSH libraries were also constructed by using 2 sets of bulks prepared from the RNA of root tissues from selected recombinant inbred lines (RILs) (10 each) for the extreme high and low root biomass phenotype. A total of 3062 unigenes (638 contigs and 2424 singletons), 51.4% of which were novel in chickpea, were derived by cluster assembly and sequence alignment of 5949 ESTs. Only 2185 (71%) unigenes showed significant BLASTX similarity (<1E-06) in the NCBI non-redundant (nr) database. Gene ontology functional classification terms (BLASTX results and GO term), were retrieved for 2006 (92.0%) sequences, and 656 sequences were further annotated with 812 Enzyme Commission (EC) codes and were mapped to 108 different KEGG pathways. In addition, expression status of 830 unigenes in response to terminal drought stress was evaluated using macro-array (dot blots). The expression of few selected genes was validated by northern blotting and quantitative real-time PCR assay. Our study compares not only genes that are up- and down-regulated in a drought-tolerant genotype under terminal drought stress and a drought susceptible genotype but also between the bulks of the selected RILs exhibiting extreme phenotypes. More than 50% of the genes identified have been shown to be associated with drought stress in chickpea for the first time. This study not only serves as resource for marker discovery, but can provide a better insight into the selection of candidate genes (both up- and downregulated) associated with drought tolerance. These results can be used to identify suitable targets for manipulating the drought-tolerance trait in chickpea.

94 citations

References
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Journal ArticleDOI
TL;DR: Protective metabolic adaptations alter physiological reactions of the whole plant, and Paramount among the mechanisms are oxygen radical scavenging, maintenance of ion uptake and water balance, and reactions altering carbon and nitrogen allocation, such that reducing power is defused.

350 citations


"Isolation and Characterization of a..." refers background in this paper

  • ...MIPS is the first enzyme in a metabolic pathway to D-pinitol, which is a cyclic sugar alcohol involved in the tolerance of drought stress that accumulates to higher concentrations in salt-tolerant legumes (Bohnert and Sheveleva, 1998; Bray et al., 2000)....

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Journal ArticleDOI
Xuetong Shen1, Hua Xiao1, Ryan T. Ranallo1, Wei Hua Wu1, Carl Wu1 
03 Jan 2003-Science
TL;DR: It is demonstrated that mutations in genes encoding inositol polyphosphate kinases that produce IP4, IP5, and IP6 impair transcription in vivo, and this results provide a link between inositl polyph phosphates, chromatin remodeling, and gene expression.
Abstract: Eukaryotes use adenosine triphosphate (ATP)–dependent chromatin-remodeling complexes to regulate gene expression. Here, we show that inositol polyphosphates can modulate the activities of several chromatin-remodeling complexes in vitro. Inositol hexakisphosphate (IP 6 ) inhibits nucleosome mobilization by NURF, ISW2, and INO80 complexes. In contrast, nucleosome mobilization by the yeast SWI/SNF complex is stimulated by inositol tetrakisphosphate (IP 4 ) and inositol pentakisphosphate (IP 5 ). We demonstrate that mutations in genes encoding inositol polyphosphate kinases that produce IP 4 , IP 5 , and IP 6 impair transcription in vivo. These results provide a link between inositol polyphosphates, chromatin remodeling, and gene expression.

349 citations


"Isolation and Characterization of a..." refers background in this paper

  • ...In addition, myo-inositol polyphosphates participate in chromatin remodelling, gene expression and mRNA export (Odom et al., 2000; Shen et al., 2003)....

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Journal ArticleDOI
TL;DR: The stereospecificity of the six hydroxyls on the inositol ring provides the basis for the functional diversity of the phosphorylated isomers that generate a selective means of intracellular and intercellular communication for coordinating cell growth.

276 citations


"Isolation and Characterization of a..." refers background in this paper

  • ...…a precursor to compounds connected to essential cellular functions, such as phosphorus storage, signal transduction, actin remodelling, membrane trafficking, stress protection, hormonal homoeostasis and cell wall biosynthesis (Loewus and Murthy, 2000; Stevenson et al., 2000; Downes et al., 2005)....

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  • ...Inositol phospholipids play a vital role in membrane trafficking and signalling pathways, auxin storage and transport, phytic acid biosynthesis, cell wall biosynthesis and production of stress-related molecules (Loewus, 1990; Loewus and Murthy, 2000; Stevenson et al., 2000; Downes et al., 2005)....

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Journal ArticleDOI
TL;DR: Functional Aspects of the MI Oxidation Pathway, Biosynthesis of Phytate, and Hydrolysis ofPhytate with reference to its MI COMPONENT are described.
Abstract: INTRODUCTION 137 BIOSYNTHESIS OF MI-I-P AND MI 1 38 1L-MI-1-P Synthase, EC 5.5.1.4 ...... ... 138 MI Monophosphatase, EC 3.1.3.25 142 Free MI andMI Kinase, EC 2.7.1.64 143 THE MI OXIDATION PATHWAy 144 MI Oxygenase, EC 1.1 3. 99.1 145 D-Glucuronokinase, EC 2. 7. 1.43 145 D-Glucuronate 1-P Uridylyltransferase, EC 2.7. 7.44 146 Metabolism of UDPGlucuronate and Gluconeogenesis 146 Functional Aspects of the MI Oxidation Pathway .... 146 PHYTATE METABOLISM WITH REFERENCE TO ITS MI COMPONENT 147 Biosynthesis of Phytate ....... .... 148 Hydrolysis of Phytate (The Phytases, EC 3.J.3.X) .... 148 MI METABOLISM IN WHEAT ........ . .. .. .. .. .. .. . 1 50 PHYSIOLOGICAL IMPLICATIONS OF MI METABOLISM . . . 1 5 1 Sugar Transport 1 5 1 Environmental Stress ........ 1 5 1 CONCLUDING REMARKS .. 152

268 citations

Journal ArticleDOI
TL;DR: Radio tracer experiments indicate that the supply ofmyo-inositol to the reaction, which converts UDP-galactose and myo- inposol to galactinol is a controlling factor in the conversion of total carbohydrate into the raffinosaccharides in both wild-type and mutant lines.
Abstract: A single, recessive mutation in soybean (Glycine max L. Merr.), which confers a seed phenotype of increased inorganic phosphate, decreased phytic acid, and a decrease in total raffinosaccharides, has been previously disclosed (S.A. Sebastian, P.S. Kerr, R.W. Pearlstein, W.D. Hitz [2000] Soy in Animal Nutrition, pp 56-74). The genetic lesion causing the multiple changes in seed phenotype is a single base change in the third base of the codon for what is amino acid residue 396 of the mature peptide encoding a seed-expressed myo-inositol 1-phospate synthase gene. The base change causes residue 396 to change from lysine to asparagine. That amino acid change decreases the specific activity of the seed-expressed myo-inositol 1-phosphate synthase by about 90%. Radio tracer experiments indicate that the supply of myo-inositol to the reaction, which converts UDP-galactose and myo-inositol to galactinol is a controlling factor in the conversion of total carbohydrate into the raffinosaccharides in both wild-type and mutant lines. That same decrease in myo-inositol 1-phosphate synthetic capacity leads to a decreased capacity for the synthesis of myo-inositol hexaphosphate (phytic acid) and a concomitant increase in inorganic phosphate.

258 citations


"Isolation and Characterization of a..." refers background in this paper

  • ...In addition, data from 15 EST libraries showed that GmMIPS1 is the preferred gene of the seed, although it is expressed in other tissue types, while GmMIPS2 is expressed in many tissues but not in developing seeds (Hitz et al., 2002)....

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  • ...The inositol phosphate biosynthesis pathway in developing seeds is poorly understood (Hitz et al., 2002; Shi et al., 2005)....

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