Regulation of nitrite reductase by light and nitrate in the cotyledons of hot pepper (Capsicum annuum L.).
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Light and nitrate are the major factors regulating the nitrite reductase (NiR) amongst various environmental and metabolic cues in plants. Hot pepper was used to investigate this regulatory mechanism of the NiR gene expression and its dependency on light and nitrate. The major results from this study are: (I) the nir partial clone (581 bp) obtained from hot pepper genomic DNA by degenerative polymerase chain reaction exhibited an amino acid sequence that is highly homologous with other plants. (II) Genomic DNA blot analysis and the NiR electrophoretic assay revealed that a small multigene family encodes NiR, which exists at least in two isoforms. (III) The light-mediated increase of NiR activity is correlated with the nitrate concentration, showing saturation kinetics above 50 mM of nitrate. (IV) Exogenous nitrate was required for the appearance of nir transcripts, but not for the enzyme activity. These results suggest that the gene expression of NiR in hot pepper is determined by the presence of nitrate at the transcriptional level. Furthermore, light has a synergistic effect on the action of nitrate on NiR levels. (+info)
Genome mapping in capsicum and the evolution of genome structure in the solanaceae.
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We have created a genetic map of Capsicum (pepper) from an interspecific F2 population consisting of 11 large (76.2-192.3 cM) and 2 small (19.1 and 12.5 cM) linkage groups that cover a total of 1245.7 cM. Many of the markers are tomato probes that were chosen to cover the tomato genome, allowing comparison of this pepper map to the genetic map of tomato. Hybridization of all tomato-derived probes included in this study to positions throughout the pepper map suggests that no major losses have occurred during the divergence of these genomes. Comparison of the pepper and tomato genetic maps showed that 18 homeologous linkage blocks cover 98.1% of the tomato genome and 95.0% of the pepper genome. Through these maps and the potato map, we determined the number and types of rearrangements that differentiate these species and reconstructed a hypothetical progenitor genome. We conclude there have been 30 breaks as part of 5 translocations, 10 paracentric inversions, 2 pericentric inversions, and 4 disassociations or associations of genomic regions that differentiate tomato, potato, and pepper, as well as an additional reciprocal translocation, nonreciprocal translocation, and a duplication or deletion that differentiate the two pepper mapping parents. (+info)
Catalytic properties of an expressed and purified higher plant type zeta-carotene desaturase from Capsicum annuum.
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The zeta-carotene desaturase from Capsicum annuum (EC 1.14.99.-) was expressed in Escherichia coli, purified and characterized biochemically. The enzyme acts as a monomer with lipophilic quinones as cofactors. Km values for the substrate zeta-carotene or the intermediate neurosporene in the two-step desaturation reaction are almost identical. Product analysis showed that different lycopene isomers are formed, including substantial amounts of the all-trans form, together with 7,7',9,9'-tetracis prolycopene via the corresponding neurosporene isomers. The application of different geometric isomers as substrates revealed that the zeta-carotene desaturase has no preference for certain isomers and that the nature of the isomers formed during catalysis depends strictly on the isomeric composition of the substrate. (+info)
Expression of the Bs2 pepper gene confers resistance to bacterial spot disease in tomato.
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The Bs2 resistance gene of pepper specifically recognizes and confers resistance to strains of Xanthomonas campestris pv. vesicatoria that contain the corresponding bacterial avirulence gene, avrBs2. The involvement of avrBs2 in pathogen fitness and its prevalence in many X. campestris pathovars suggests that the Bs2 gene may be durable in the field and provide resistance when introduced into other plant species. Employing a positional cloning strategy, the Bs2 locus was isolated and the gene was identified by coexpression with avrBs2 in an Agrobacterium-mediated transient assay. A single candidate gene, predicted to encode motifs characteristic of the nucleotide binding site-leucine-rich repeat class of resistance genes, was identified. This gene specifically controlled the hypersensitive response when transiently expressed in susceptible pepper and tomato lines and in a nonhost species, Nicotiana benthamiana, and was designated as Bs2. Functional expression of Bs2 in stable transgenic tomatoes supports its use as a source of resistance in other Solanaceous plant species. (+info)
Resistance of Capsicum annuum 'Avelar' to pepper mottle potyvirus and alleviation of this resistance by co-infection with cucumber mosaic cucumovirus are associated with virus movement.
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Capsicum annuum cv. Avelar plants resist systemic infection by the Florida isolate of pepper mottle potyvirus (PepMoV-FL). Immuno-tissue blot analysis for detection of PepMoV-FL infection in selected stem segments revealed that virus moved down the stem in external phloem, and, over time, accumulated to detectable levels throughout stem sections (appearing to accumulate in external and internal phloem) taken from below the inoculated leaf. At 21 days post-inoculation, PepMoV-FL was detected in stem segments one or two internodes above the inoculated leaf; however, no virus was observed in internal phloem in stem segments beyond these internodes. In contrast to these observations, PepMoV-FL was detected in the internal phloem of all internodes of the stem located above the inoculated leaf, with subsequent movement into non-inoculated leaves, in Avelar plants co-infected with PepMoV-FL and cucumber mosaic cucumovirus (CMV-KM). No apparent enhancement of PepMoV-FL accumulation occurred in protoplasts inoculated with PepMoV-FL alone versus a mixed inoculum of PepMoV-FL and CMV-KM. These findings confirm earlier observations that potyvirus movement up the stem of Capsicum species occurs via internal phloem. It is also shown that PepMoV-FL does not accumulate to detectable levels in internal phloem in the stem of Avelar plants, thereby limiting its movement to within the inoculated leaf and lower portions of the stem; however, co-infection of Avelar plants with CMV-KM alleviates this restricted movement, allowing PepMoV-FL to invade young tissues systemically. (+info)
A cytochrome P450 gene is differentially expressed in compatible and incompatible interactions between pepper (Capsicum annuum) and the anthracnose fungus, Colletotrichum gloeosporioides.
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The anthracnose fungus, Colletotrichum gloeosporioides, was previously shown to have an incompatible interaction with ripe-red fruit of pepper (Capsicum annuum). However, the fungus had a compatible interaction with unripe-mature-green fruit. Using mRNA differential display, we isolated and characterized a PepCYP gene expressed in the incompatible interaction. The PepCYP gene encodes a protein homologous to cytochrome P450 proteins containing a heme-binding domain. The expression level of PepCYP is higher in the incompatible interaction than in the compatible interaction, and then remains elevated in the incompatible interaction. In the compatible interaction, the expression of PepCYP is transient. The induction of PepCYP gene is up-regulated by wounding or jasmonic acid treatment during ripening. Analysis of PepCYP expression by in situ hybridization shows that the accumulation of PepCYP mRNA is localized in the epidermal cell layers, but not in the cortical cell layers. An examination of transverse sections of the fruits inoculated with the fungus shows that the fungus invades and colonizes the epidermal cell layers of the unripe fruit at 24 and 72 h after inoculation, respectively, but not those of the ripe fruit. These results suggest that the PepCYP gene product plays a role in the defense mechanism when the fungus invades and colonizes the epidermal cells of fruits in the incompatible interaction during the early fungal infection process. (+info)
Biosynthesis of terpenoids: YchB protein of Escherichia coli phosphorylates the 2-hydroxy group of 4-diphosphocytidyl-2C-methyl-D-erythritol.
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A comparative analysis of all published complete genomes indicated that the putative orthologs of the unannotated ychB gene of Escherichia coli follow the distribution of the dxs, dxr, and ygbP genes, which have been shown to specify enzymes of the deoxyxylulose phosphate pathway of terpenoid biosynthesis, thus suggesting that the hypothetical YchB protein also is involved in that pathway. To test this hypothesis, the E. coli ychB gene was expressed in a homologous host. The recombinant protein was purified to homogeneity and was shown to phosphorylate 4-diphosphocytidyl-2C-methyl-D-erythritol in an ATP-dependent reaction. The reaction product was identified as 4-diphosphocytidyl-2C-methyl-D-erythritol 2-phosphate by NMR experiments with various (13)C-labeled substrate samples. A (14)C-labeled specimen of this compound was converted efficiently into carotenoids by isolated chromoplasts of Capsicum annuum. The sequence of E. coli YchB protein is similar to that of the protein predicted by the tomato cDNA pTOM41 (30% identity), which had been implicated in the conversion of chloroplasts to chromoplasts. (+info)
Isolation, partial sequencing, and expression of pathogenesis-related cDNA genes from pepper leaves infected by Xanthomonas campestris pv. vesicatoria.
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Specific cDNAs showing differential expression in bacteria-infected pepper leaves as opposed to healthy leaves were isolated from a pepper cDNA library from hypersensitive response (HR) lesions of leaves infected with an avirulent strain of Xanthomonas campestris pv. vesicatoria. Among a total of 282 cDNA clones tested, 36 individual cDNA genes (13%) hybridized strongly or differentially to the cDNA probes from bacteria-infected leaves. Ten Capsicum Annuum-Induced (CAI) genes encoding putative thionin, lipid transfer protein I and II, osmotin (PR-5), class I chitinase, beta-1,3-glucanase, SAR 8.2, stellacyanin, leucine-rich repeat protein, and auxin-repressed protein were identified. Two CAI genes showed little or no sequence homology to the previously sequenced plant genes. Transcripts of the CAI genes were strongly or preferentially induced in pepper tissues by infection with X. campestris pv. vesicatoria or Phytophthora capsici, and by abiotic elicitor treatment. In particular, most of the CAI genes were strongly induced in pepper tissues by ethephon and methyl jasmonate. (+info)