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Role of a novel pathogen-induced pepper C3-H-C4 type RING-finger protein gene, CaRFPI, in disease susceptibility and osmotic stress tolerance.

Limited information is available about the roles of RING-finger proteins in plant defense. A pepper CaRFP1 encoding the C3-H-C4 type RING-finger protein that physically interacted with the basic PR-1 protein CABPR1 was isolated from pepper leaves infected by Xanthomonas campestris pv. vesicatoria. The CaRFP1 protein has VWFA domain, and N-terminal serine-rich and C-terminal cysteine-rich regions. The CaRFP1 transcripts accumulated earlier than did those of the basic PR-1 gene CABPR1 during the incompatible interaction of pepper leaves with X. campestris pv. vesicatoria, as well as in the systemic, uninoculated pepper leaf tissues. The CaRFP1 gene also was induced in pepper leaf tissues infected by Colletotrichum coccodes. The CaRFP1 gene was strongly induced much earlier by salicylic acid, ethylene and methyl jasmonate treatments, as well as environmental stresses including methyl viologen, mannitol and NaCl treatments. Overexpression of the CaRFP1 gene in the transgenic Arabidopsis plants conferred disease susceptibility to Pseudomonas syringae pv. tomato infection, accompanied by reduced PR-2 and PR-5 gene expression, suggesting that the CaRFP1 acts as an E3 ligase for polyubiquitination of target PR proteins. Exogenous salicylic acid treatment also abolished PR-2 and PR-5 gene expression in the transgenic plants. Differential osmotic stress tolerance was induced by high salt and drought in the CaRFPI-overexpressing plants during germination and seedling development, which was closely correlated with abscisic acid sensitivity of Arabidopsis plants. These results suggest that the CaRFP1 gene functions as an early defense regulator controlling bacterial disease susceptibility and osmotic stress tolerance.

Amino Acid Sequence↗

Evaluation of the modulatory influence of black pepper (Piper nigrum, L.) on the hepatic detoxication system.

The present paper assesses the modifying potential of black pepper on the hepatic biotransformation system in mice. The modulatory effect was assessed on glutathione S-transferase (GST), cytochrome b5 (cyt. b5), cytochrome P-450 (cyt. P-450), acid-soluble sulfhydryl (-SH) content and malondialdehyde (MDA) level. Swiss albino mice of either sex (eight weeks old) were fed on a diet containing 0.5%, 1% and 2% black pepper (w/w) for 10 and 20 days. The findings revealed a significant and dose-dependent increase in GST and -SH content in the experimental groups except the one maintained on 0.5% black pepper diet for 10 days. Elevated levels of cyt. b5 and cyt. P-450 were also statistically significant and dose-dependent. The level of MDA was lowered in the group fed on 2% black pepper diet for 20 days. Being a potential inducer of detoxication system, the possible chemopreventive role of black pepper in chemical carcinogenesis is suggested.

Animals↗

Suppression of CaCYP1, a novel cytochrome P450 gene, compromises the basal pathogen defense response of pepper plants.

A putative cytochrome P450 gene from chili pepper, Capsicum annuum L. Bukang cytochrome P450 (CaCYP1), was identified using cDNA microarray analysis of gene expression following induction of the leaf hypersensitive response by inoculation of pepper plants with the non-host pathogen Xanthomonas axonopodis pv. glycines 8ra. The full-length cDNA of CaCYP1 encoded a protein of 514 amino acid residues, which contained a putative hydrophobic membrane anchoring domain in the N-terminal region, and a heme-binding motif in the C-terminal region. Analysis of the deduced amino acid sequence of CaCYP1 revealed that it has high homology to Arabidopsis CYP89A5, the function of which is unknown. Expression of CaCYP1 was preferentially increased in pepper plants in response to non-host pathogen inoculation and also during the host resistance response. CaCYP1 expression also increased following treatment with salicylic acid and abscisic acid, while treatment with ethylene had a mild effect. Using a virus-induced gene silencing-based reverse genetics approach, we demonstrated that suppression of CaCYP1 results in enhanced susceptibility to bacterial pathogens. Interestingly, gene silencing of CaCYP1 in pepper plants resulted in the reduced expression of the defense-related genes CaLTP1, CaSIG4, and Cadhn. Our results indicated that CaCYP1, a novel cytochrome P450 in pepper plants, may play a role in plant defense response pathways that involve salicylic acid and abscisic acid signaling pathways.

Amino Acids↗

Functional study of hot pepper 26S proteasome subunit RPN7 induced by Tobacco mosaic virus from nuclear proteome analysis.

Two-dimensional gel electrophoresis (2-DE) was applied for the screening of Tobacco mosaic virus (TMV)-induced hot pepper (Capsicum annuum cv. Bugang) nuclear proteins. From differentially expressed protein spots, we acquired the matched peptide mass fingerprint (PMF) data, analyzed by MALDI-TOF MS, from the non-redundant hot pepper EST protein FASTA database using the VEMS 2.0 software. Among six identified nuclear proteins, the hot pepper 26S proteasome subunit RPN7 (CaRPN7) was subjected to further study. The level of CaRPN7 mRNA was specifically increased during incompatible TMV-P(0) interaction, but not during compatible TMV-P(1.2) interaction. When CaRPN7::GFP fusion protein was targeted in onion cells, the nuclei had been broken into pieces. In the hot pepper leaves, cell death was exacerbated and genomic DNA laddering was induced by Agrobacterium-mediated transient overexpression of CaPRN7. Thus, this report presents that the TMV-induced CaRPN7 may be involved in programmed cell death (PCD) in the hot pepper plant.

Amino Acid Sequence↗

Effect of different Thai traditional processing of various hot chili peppers on urethane-induced somatic mutation and recombination in Drosophila melanogaster: assessment of the role of glutathione transferase activity.

Four different Thai traditional chili peppers, namely bird pepper (Capsicum frutescens), red chili spur peppers (Capsicum annuum), green bell peppers and sweet pepper (C. annuum) were investigated for their antimutagenic properties. Each chili was prepared in three formulations commonly used for chili food processing; raw paste (chili ground in water), pickled in vinegar or stir-fried in palm oil. Each sample was tested for its antimutagenic effect against urethane by using the somatic mutation and recombination of wing hair of Drosophila melanogaster as an indicator. Three-day-old larvae, trans-heterozygous for two genetic markers, multiple wing hairs mwh and orrigon (ORR;flr3), were exposed to urethane alone or in combination with each chili formulation. The various processing methods for chilies differentially extracted the antimutagenic chili components. The specific chili as well as the method of processing influenced the observed antimutagenic properties against urethane. This suggested each chili contains a unique complex mixture of many antimutagens. Co-treatment and pre-treatment experiments showed that both direct and indirect protective mechanisms are involved in an 'activation' process to give antimutagenesis effects. An association between antigenotoxicity and glutathione transferase activity could not be established.

Animals↗

Effect of composted sewage sludge application to soil on sweet pepper crop (Capsicum annuum var. annuum) grown under two exploitation regimes.

The aim of this study is to monitor the effect of the application of three increasing amounts of composted sewage sludge (3, 6 and 9 kg compost m(-2)) on the physico-chemical properties of a horticultural calcareous soil where sweet pepper plants (Capsicum annuum var. annuum) cv. California were grown. A comparative study of two different exploitation regimes was carried out; the first was an open-air field-grown plot and the second plot was kept under controlled conditions in a greenhouse. Changes in physical and chemical properties measured in soil and sweet pepper crop were recorded during crop growth in order to measure the evolution of these properties as a consequence of increasing compost applications. Organic matter, total nitrogen Kjeldahl and available phosphorus contents increased in soil after composted sewage sludge applications. The 9 kg compost m(-2) application promoted the appearance of deleterious effects on the properties of soil, such as salt accumulation, a significant increase in the electrical conductivity and an input of heavy metals (Pb>Cr>Cd). The 6 kg compost m(-2) application provided a supply of nutrients necessary to grow peppers plants under both exploitation regimes. Pepper fruit biomass production under greenhouse was almost 60% higher compared to that of the open-air plot. Lower contents of Ca and increased levels of Cu in fruit under greenhouse growing conditions compared to those of open-air grown peppers seemed to promote the occurrence of blossom-end rot, affecting more than 10% of the harvested fruits.

Biodegradation, Environmental↗

Toxic carriers in pepper sprays may cause corneal erosion.

We describe four patients who developed corneal erosion after an exposure to a pepper spray containing toxic carriers. Two of these patients were exposed to a pepper gas containing 5% oleoresin capsicum (OC) as an irritant and 92% trichlorethylene or unknown amount of dichloromethane as a carrier. One patient was exposed to a mock (containing 92% trichlorethylene as a carrier) training pepper gas without OC. The fourth patient was exposed to an unidentified Russian pepper gas spray. Two of the patients were examined by in vivo confocal microscopy to demonstrate the depth and quality of the stromal damage. To test the toxicity of the commercial tear spray, it was analyzed and test sprayed on a soft contact lens and into a plastic cup. Visual acuity was measured and the eyes were examined with a slit-lamp up to 5 months. Physical damage to a soft contact lens was visually acquired. All patients showed a long-lasting, deep corneal and conjuctival erosion, which resolved partly with medical therapy during the following weeks/months. Confocal microscopy revealed corneal nerve damage, and keratocyte activation reaching two-thirds of stroma for one patient. The spray caused serious damage to both the soft contact lens and the plastic cup. The safety of the commercially available pepper sprays should be assessed before marketing, and a list of acceptable ingredients created. The sprays should also have instructions on the use of the compound as well as on the first aid measures after the exposure. Solvents known to be toxic should not be used.

Adolescent↗

Capsaicin pepper, cancer and ethnicity.

The 'hot' sensation produced by exposure to pepper is apparently due to two natural carcinogens: capsaicin in chili type peppers and safrole in black/white pepper. There are four cookeries in the United States that are noted for their high pepper content: Mexican-American, Cajun, white Creole, and black Creole. Each is largely confined to a single ethnic-cultural group which is concentrated in some counties. By use of county population and mortality data, significantly higher rates for stomach and liver cancer were found in counties inhabited by these four ethnic-cultural groups than in matched control counties. This involved both sexes. The cancer increase was dependent on the concentration of these groups in a county. These results strengthen and extend an earlier case-control study which found odds ratios above 5 for the stomach cancer association with capsaicin pepper. It is further evidence that capsaicin is a human carcinogen.

Capsaicin↗

Corneal abrasions associated with pepper spray exposure.

Pepper spray containing oleoresin capsicum is used by law enforcement and the public as a form of nonlethal deterrent. Stimulated by the identification of a case of a corneal abrasion associated with pepper spray exposure, a descriptive retrospective review of a physician-maintained log of patients presenting to a jail ward emergency area over a 3-year period was performed. The objective was to give some quantification to the frequency with which an emergency physician could expect to see corneal abrasions associated with pepper spray exposure. Of 100 cases of pepper spray exposure identified, seven patients had sustained corneal abrasions. We conclude that corneal abrasions are not rare events when patients are exposed to pepper spray and that fluorescein staining and slit lamp or Wood's lamp examination should be performed on all exposed patients in whom corneal abrasions cannot be excluded on clinical grounds.

Adolescent↗

Antimutagenic effect of one variety of green pepper (Capsicum spp.) and its possible interference with the nitrosation process.

It is known that the poblano green pepper, a significant component in the Mexican diet, contains certain natural compounds such as chlorophyll, beta-carotene, and vitamins, which have antimutagenic and/or anticarcinogenic properties. Using the somatic mutation and recombination test in wing cells of Drosophila melanogaster, an extract of the poblano pepper (Capsicum spp.) was evaluated to determine its antimutagenic effect against the nitrosation process, simulating the process occurring in the human stomach caused by known food additives. Larvae of 72h old D. melanogaster of standard (ST) and high bioactivation (HB) crosses were exposed in a simultaneous, chronic treatment with the juice expressed from the crushed, whole, fresh pepper fruit, plus the mixture of 20mM methyl urea (MU) and sodium nitrite (SN), mixed with the animals' food. Three doses of pepper juice (12.5, 25, and 50%) were used. The background mutation rate given as spots per wing was 0.36 and 0.48 for ST and HB, respectively. Mutation frequencies produced by the MU and SN mixture was 1.73 (ST) and 26.46 (HB) mutations per wing. The poblano juice decreased the above rates between 40 and 80%, respectively. The experiments suggest that some compounds present in the green pepper may cause this antimutagenic effect by interfering with the nitrosation process. The role of the extract and one of its components, such as vitamin C, in the nitrosation process will be discussed.

Animals↗

Characterization of odorants causing an atypical aroma in white pepper powder (Piper nigrum L.) based on quantitative measurements and orthonasal breakthrough thresholds.

Application of the aroma extract dilution analysis on an extract of white pepper powder showing an intense fecal, cowshed-like off-flavor revealed 3-methylindole (fecal, swine-manure) and 4-methylphenol (fecal, horse-like) with the highest flavor dilution (FD) factors among the 22 odor-active compounds detected. In addition, high FD factors and/or undesirable odor qualities suggested 3-methylphenol (phenolic), butanoic acid (cheese-like), and 2- and 3-methylbutanoic acid (cheese-like) as well as pentanoic acid and hexanoic acid (cheese-like odors) as contributors to the malodor. Although the intensities of the off-note were clearly different in 50 commercial samples of white pepper, quantitation of 3-methylindole and 3- and 4-methylphenol as well as of the five short-chain acids by means of stable isotope dilution assays showed similar concentrations in most of the samples. Storage of a freshly ground white pepper powder for up to 7 months revealed a significant decrease in the typical odor qualities of white pepper and an increase in the fecal odor note with storage time. Because the concentrations of the odorants mentioned above were not much changed during storage, possibly very volatile odorants, such as alpha-pinene, which are able to mask the malodor, are lost during storage of, in particular, pepper powders. On the basis of odor activity values, which were calculated using breakthrough thresholds, in particular, 3-methylindole, 4-methylphenol, 3-methylphenol, and butanoic acid could be suggested as the main sources of the fecal off-flavor.

Animals↗

Effect of capsaicin-containing red pepper sauce suspension on upper gastrointestinal motility in healthy volunteers.

Afferent nerves play a major role in the regulation of gastrointestinal motility. The questions remains if specific food ingredients can selectively activate such fibers. The aim of the study was to investigate the effect of intraesophageal application of a capsaicin-containing red pepper sauce (Tabasco) suspension on upper gastrointestinal motility in a controlled trial. After a baseline recording [esophageal motility, balloon distension, electrogastrogram (EGG)], red pepper or saline solution was infused intraesophageally in seven healthy volunteers. At 30 min gastric emptying and orocecal transit time were determined using a [13C]acetate and H2-lactulose breath test. Infusion of red pepper sauce suspension significantly increased the amplitudes (65.8 +/- 3 to 78.5 +/- 4.7 mm Hg, P < 0.05) and propagation velocity (2.9 +/- 0.3 to 4.25 +/- 0.3 sec, P < 0.05) of esophageal pressure waves and LES pressure (17.8 +/- 1.4 to 23.7 +/- 2.6 mm Hg, P < 0.05). It significantly decreased perception and discomfort threshold of intraesophageal balloon distension, reduced the percentage of normal electrical activity in the EGG, and delayed gastric emptying (saline: T(1/2) 42.9 +/- 12.0 min vs red pepper: T(1/2) 66.8 +/- 19.0 min, P < 0.05). Despite the prolongation of gastric emptying, orocecal transit time was not altered, indicating an actual increase of intestinal transit. Esophageal application of capsaicin-containing red pepper sauce suspension had profound changes on upper gastrointestinal motility, which could improve clearance and protection of the esophagus and could lead to retention of the irritant in the stomach and faster transit through the small bowel.

Adult↗

The treatment of functional dyspepsia with red pepper.

AIM: : To decrease the intensity of dyspeptic symptoms by impairing the visceral nociceptive C-type fibres with capsaicin, contained in red pepper powder. METHODS: : The study was performed on 30 patients with functional dyspepsia and without gastro-oesophageal reflux disease and irritable bowel syndrome. After a 2-week washout period, 15 patients received, before meals randomly and in a double-blind manner, 2.5 g/day of red pepper powder for 5 weeks, and 15 patients received placebo. A diary sheet was given to each patient to record, each day, the scores of individual and overall symptom intensity, which subsequently were averaged weekly and over the entire treatment duration. RESULTS: : The overall symptom score and the epigastric pain, fullness and nausea scores of the red pepper group were significantly lower than those of the placebo group, starting from the third week of treatment. The decrease reached about 60% at the end of treatment in the red pepper group, whilst placebo scores decreased by less than 30%. CONCLUSIONS: : Red pepper was more effective than placebo in decreasing the intensity of dyspeptic symptoms, probably through a desensitization of gastric nociceptive C-fibres induced by its content of capsaicin. It could represent a potential therapy for functional dyspepsia.

Adult↗

Fructokinase and hexokinase from pollen grains of bell pepper (Capsicum annuum L.): possible role in pollen germination under conditions of high temperature and CO2 enrichment.

The processes of pollen grain development and germination depend on the uptake and metabolism of pollen sugars. In pepper (Capsicum annuum L.), initial sugar metabolism includes sucrose hydrolysis by invertase and subsequent phosphorylation of glucose and fructose by hexose kinases. The main objective of this study was to investigate changes in fructokinase (EC 2.7.1.4) and hexokinase (EC.2.7.1.1) activities in pepper flowers during their development, and to study the possible roles of these enzymes in determining pollen germination capacity under high temperature and under CO(2) enrichment, previously shown to modify sugar concentrations in pepper pollen (Aloni et al., 2001 Physiologia Plantarum 112: 505-512). Fructokinase (FK) activity was predominant in pepper pollen, and increased during pollen maturation. Pollen hexokinase (HK) activity was low and did not change throughout pollen development. High-temperature treatment (day/night, 32/26 degrees C) of pepper plants reduced the percentage of pollen that germinated compared with that under normal temperatures (26/22 degrees C), and concomitantly reduced the activity of FK in mature pollen. High temperature also reduced FK and HK activity in the anther. Under high ambient CO(2) (800 micro l l(-1)) pollen FK activity was enhanced. The results suggest that pollen and anther FK may play a role in the regulation of pollen germination, possibly by providing fructose-6-phosphate for glycolysis, or through conversion to UDP-glucose (UDPG) to support the biosynthesis of cell wall material for pollen tube growth. High temperature stress and CO(2) enrichment may influence pollen germination capacity by affecting these pathways.

Calcium Chloride↗

Managing the pepper maggot (Diptera: Tephritidae) using perimeter trap cropping.

A perimeter trap crop barrier of hot cherry peppers, border-row insecticide applications, and a combination of the two management strategies were evaluated to see if they could protect a centrally located main crop of bell peppers from oviposition and infestation by the pepper maggot, Zonosemata electa (Say). In large plots, the main cash crop of bell peppers was protected from the majority of the oviposition and infestation by all three barriers. The combination sprayed/trap crop barrier provided the best protection against both oviposition and infestation and resulted in over 98% pest-free fruit at harvest. Maggots infested only 1.7% of the main crop fruit when protected by a sprayed or unsprayed trap crop barrier, compared with 15.4% in control plots. The perimeter sprayed/trap crop strategy was employed in three commercial fields in 2000 and 2001. The combination barrier resulted in superior insect control and reduced insecticide use at all commercial locations, compared with the same farms' past history or to farms using conventional and integrated pest management (IPM) methods. Economic analysis showed that the technique is more cost effective and profitable than relying on whole-field insecticide applications to control the pepper maggot. Farmer users were surveyed and found the perimeter trap crop technique simple to use, with many hard-to-measure benefits associated with worker protection issues, marketing, personnel/management relations, pest control and the environment. Use of the perimeter trap crop technique as part of an IPM or organic program can help improve crop quality and overall farm profitability, while reducing pesticide use and the possibility of secondary pest outbreaks.

Agriculture↗

Chili pepper consumption and gastric cancer in Mexico: a case-control study.

Laboratory studies indicate that capsaicin, the hot-tasting component of chili peppers, may be carcinogenic. A population-based case-control study was conducted in Mexico City during 1989-1990 to evaluate the relation between chili pepper consumption and gastric cancer risk. The study included 220 incident cases and 752 controls randomly selected from the general population. Information was collected by interview. Chili pepper consumers were at high risk for gastric cancer compared with nonconsumers (age- and sex-adjusted odds ratio = 5.49, 95% confidence interval (CI) 2.72-11.06). Among consumers, there was a highly significant trend of increasing risk with increasing self-rated level of consumption (low, medium, and high) (p = 2 x 10(-7). The odds ratio for high-level consumers compared with nonconsumers was 17.11 (95% CI 7.78-37.59). However, when consumption was measured as frequency per day, a significant trend among consumers was not observed. Multivariable adjustment increased the magnitude of the chili pepper-gastric cancer association, but a significant trend among consumers (measured as frequency per day) was still not observed. Chili pepper consumption may be a strong risk factor for gastric cancer, but further studies are needed to test this hypothesis.

Adenocarcinoma↗

Ectopic expression of Tsi1 in transgenic hot pepper plants enhances host resistance to viral, bacterial, and oomycete pathogens.

In many plants, including hot pepper plants, productivity is greatly affected by pathogen attack. We reported previously that tobacco stress-induced gene 1 (Tsi1) may play an important role in regulating stress responsive genes and pathogenesis-related (PR) genes. In this study, we demonstrated that overexpression of Tsi1 gene in transgenic hot pepper plants induced constitutive expression of several PR genes in the absence of stress or pathogen treatment. The transgenic hot pepper plants expressing Tsi1 exhibited resistance to Pepper mild mottle virus (PMMV) and Cucumber mosaic virus (CMV). Furthermore, these transgenic plants showed increased resistance to a bacterial pathogen, Xanthomonas campestris pv. vesicatoria and also an oomycete pathogen, Phytophthora capsici. These results suggested that ectopic expression of Tsi1 in transgenic hot pepper plants enhanced the resistance of the plants to various pathogens, including viruses, bacteria, and oomycete. These results suggest that using transcriptional regulatory protein genes may contribute to developing broad-spectrum resistance in crop plants.

Bacteria↗

Wound-inducible proteinase inhibitors in pepper. Differential regulation upon wounding, systemin, and methyl jasmonate.

Seven small (approximately 6,000 D) wound-inducible proteinase inhibitor proteins were isolated from leaves of pepper (Capsicum annuum) plants that are members of the potato inhibitor II family. N-terminal sequences obtained indicated that the pepper leaf proteinase inhibitors (PLPIs) exhibit homology to two GenBank accessions that code for preproteins containing three isoinhibitors domains each that, when post-translationally processed, can account for the mixture of isoinhibitors that are reported herein from pepper leaves. A constitutive level of PLPI proteins was found in pepper leaves, and these levels increased up to 2.6-fold upon wounding of the lower leaves. Exposing intact plants to methyl jasmonate vapors induced the accumulation of PLPIs. Supplying excised young pepper plants with water through the cut stems induced PLPI proteins to levels higher than those found in intact plants, but with high variability. Supplying the excised plants with systemin did not result in an increase of PLPI levels that were statistically higher than levels found in excised plants. Gel-blot analyses of PLPI induction revealed the presence of two mRNA bands, having slightly different mobilities in agarose gels. Only the low M(r) mRNA is present in untreated control plants, and it appears to be responsible for the constitutive levels of PLPI found in leaves. Both mRNA species are wound- and methyl jasmonate-inducible. Only the low- M(r) species is weakly induced by systemin, indicating a differential expression of the two PLPI species.

Acetates↗