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Reduction of listeria monocytogenes on green peppers (Capsicum annuum L.) by gaseous and aqueous chlorine dioxide and water washing and its growth at 7 degrees C.

Reduction of Listeria monocytogenes Scott A on uninjured and injured surfaces of green peppers after 0.3- and 3-mg/ liter gaseous and aqueous ClO2 treatment and water washing for 10 min at 20 degrees C was studied. Growth of the L. monocytogenes untreated or treated with 0.6 mg/liter ClO2 gas for 30 min at 20 degrees C on green peppers also was investigated. A membrane-surface-plating method was used for resuscitation and enumeration of L monocytogenes treated with ClO2. The bacterial viability on pepper surfaces was visualized using confocal laser scanning microscopy (CLSM). Live and dead cells of L. monocytogenes were labeled with a fluorescein isothiocyanate-labeled antibody and propidium iodide, respectively. More than 6 log CFU/5 g L. monocytogenes on uninjured surfaces and about 3.5 log CFU/5 g on injured surfaces were inactivated by both 3-mg/liter and 0.6-mg/liter ClO2 gas treatments. The 3-mg/liter aqueous ClO2 treatment achieved 3.7- and 0.4-log reductions on uninjured and injured surfaces, respectively; whereas, water washing alone showed 1.4- and 0.4-log reductions, respectively. ClO2 gas treatment was the most effective in reducing L. monocytogenes on both uninjured and injured green pepper surfaces, when compared with aqueous ClO2 treatment and water washing. The significant difference (P < 0.05) between log reductions on uninjured and injured surfaces and the results from CLSM analysis suggested that injured surfaces protected more bacteria from sanitation treatments than did uninjured surfaces. Not only could L. monocytogenes grow on green pepper surfaces at 7 degrees C, bacteria that survived the 0.6-mg/liter ClO2 gas treatment also could grow.

Capsicum↗

Response surface modeling for the inactivation of Escherichia coli O157:H7 on green peppers (Capsicum annuum L.) by chlorine dioxide gas treatments.

The effects of chlorine dioxide (ClO2) gas concentration (0.1 to 0.5 mg/liter), relative humidity (RH) (55 to 95%), treatment time (7 to 135 min), and temperature (5 to 25 degrees C) on inactivation of Escherichia coli O157:H7 on green peppers were studied using response surface methods. A four-factor, central, composite, rotatable design was used. The microbial log reduction was measured as a response. A direct membrane-surface-plating method with tryptic soy agar and sorbitol MacConkey agar was used to resuscitate and enumerate ClO2-treated E. coli O157:H7 cells. The statistical analysis and the predictive model developed in this study suggest that ClO2 gas concentration, treatment time, RH, and temperature all significantly (P < 0.01) increased the inactivation of E. coli O157:H7. ClO2 gas concentration was the most important factor, whereas temperature was the least significant. The interaction between ClO2 gas concentration and RH indicated a synergistic effect. The predictive model was validated, and it could be used to determine effective ClO2 gas treatments to achieve a 5-log reduction of E. coli O157:H7 on green peppers.

Capsicum↗

Regulation of nitrite reductase by light and nitrate in the cotyledons of hot pepper (Capsicum annuum L.).

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.

Amino Acid Sequence↗

Constitutive expression of rice MADS box gene using seed explants in hot pepper (Capsicum annuum L.).

Two transgenic pepper plants were obtained from 255 seed explants that were infected with Agrobacterium LBA4404 (pGA1209). One of them (PT2) showed morphological change, such as dwarfism and early flowering by the constitutive expression of the rice OsMADS1 gene. The in vitro condition of the plant regeneration has been optimized from hypocotyl explants on a MS medium that was supplemented with zeatin 3 mg/L, IAA 0.3 mg/L for shoot induction. The optimal rooting condition was at NAA 0.3 mg/L. The transformation frequency was 0.8% from the total hypocotyls. DNA and RNA hybridization analyses showed that the introduced gene was integrated and stably expressed in regenerated plants.

Capsicum↗

Application of RAPD and SCAR markers for purity testing of F1 hybrid seed in chili pepper (Capsicum annuum).

A simpler and better method for purity testing of hybrid pepper seed was developed. The simplest method for extracting genomic DNA, the NaOH method, was chosen. Two RAPD markers identifying male and female parents were also developed, and the PCR products of male- and female-specific RAPD markers were cloned and sequenced. From these sequences, new longer primers were constructed for conversion into SCAR markers. In blind tests the RAPD and SCAR markers were able to reliably detect contaminating exotic seeds. These PCR-based markers are therefore directly applicable for purity testing by seed companies. In addition, the PCR products of the SCAR markers could be identified by direct staining methods such as ethidium bromide and pellet painting without electrophoresis.

Capsicum↗

Cold-inducible transcription factor, CaCBF, is associated with a homeodomain leucine zipper protein in hot pepper (Capsicum annuum L.).

C-Repeat/drought responsive element binding factor (CBF1/DREB1b) is a well known transcriptional activator that is induced at low temperature and in turn induces the CBF regulon (CBF-targeted genes). We have cloned and characterized two CBF1-like cDNAs, CaCBF1A and CaCBF1B, from hot pepper. CaCBF1A and CaCBF1B were not produced in response to mechanical wounding or abscisic acid but were induced by low-temperature stress at 4 degrees . When plants were returned to 25 degrees , their transcript levels of the CBF1-like genes decreased markedly within 40 min. Long-term exposure to chilling resulted in continuous expression of these genes. The critical temperature for induction of CaCBF1A was between 10 and 15 degrees . Low temperature led to its transcription in roots, stems, leaves, fruit without seeds, and apical meristems, and a monoclonal antibody against it revealed a significant increase in CaCBF1A protein by 4 h at 4 degrees . Two-hybrid screening led to the isolation of an homeodomain leucine zipper (HD-Zip) protein that interacts with CaCBF1B. Expression of HD-Zip was elevated by low temperature and drought.

Abscisic Acid↗

[cDNA cloning and expression of a cytosolic small heat shock protein gene (CaHSP18) from Capsicum annuum].

Full length 779 bp cytosolic clsss I sHSP cDNA was cloned from heat-shocked sweet pepper leaves using a PCR approach with degenerate primers designed from conserved motifs found in a number of plant cytosolic clsss I sHSP genes, named CaHSP18. Its accession number is AY284925. CaHSP18 was high identified with Nicotiana tabacum (Fig.1). A multi-gene family was found in sweet pepper genomic DNA by Southern-blot analysis (Fig.3). Northern blot revealed that the expression of CaHSP18 in sweet pepper was induced by heat treatment and was significantly different between different tissues (Fig.4). The transcription of CaHSP18 in leaves and stems were higher than those in roots. The expression of CaHSP18 could be detected after chilling treatment at 4 degrees C for 2 d. Recombinant CaHSP18 was overexpressed in Escherichia coli to study its possible function under heat and chilling stress. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of cell lysates suggested that CaHSP18 was expressed in Escherichia coli (Fig.6). The growth of wild type and transformed cells was similar at 37 degrees C (Fig. 5). Upon transfer from 37 degrees C to 50 degrees C, a temperature known to cause cell autolysis, those cells that accumulated CaHSP18 showed improved viability compared with the control lines (Fig.7). To test the hypothesis that sHSPs may be involved in protection against chilling stress, the viability of recombinant cells at 4 degrees C was studied. The sweet pepper CaHSP18 significantly enhanced cell survivability (Fig.8). These results indicate that plant cytosolic clsss I sHSP have protective functions not only against heat stress but also against chilling stress.

Amino Acid Sequence↗

Effect of a mixture of red pepper (Capsicum frutescens) and an amino-acid on gastric acid secretion.

The effects of a mixture of red pepper suspension and alpha-alanine (an amino-acid) on gastric acid secretion in 33 duodenal ulcer and 18 non-duodenal ulcer patients were studied. This mixture caused an increase of gastric acid in the two groups of patients, more so in the duodenal ulcer group (0.025 > P > 0.010). Compared with the results of previous studies the administration of alpha-alanine in red pepper suspension produced less gastric acid than the fresh red pepper suspension alone.

Adult↗

Seed borne nature of Azotobacter chroococcum in chilli (Capsicum annum) and its role in seed germination and plant growth.

Investigations were carried out on seed borne nature of Azotobacter chroococcum in chilli and their role in plant growth at the Department of Agriculture Microbiology, University of Agricultural Sciences, Dharwad. Azotobacter chroococcum were isolated from both surface sterilized and unsterilized seeds of 14 varieties of chilli at different stages of fruit maturity. Inoculation with these strains increased the seed germination, root, shoot length and total dry matter content of chilli plants significantly in both sterilized and unsterilized soil., the results of which are presented.

Azotobacter↗

Qualitative analysis of MDR-reversing Anastasia Black (Russian black sweet pepper, Capsicum annuum, Solanaceae) extracts and fractions by HPLC and LC-MS-MS methods.

In earlier experiments, the MDR (multidrug resistance)-reversal activities of Anastasia Black (Russian black sweet pepper) extracts had been analysed. Recently, the most effective MDR reversing extracts and fractions have been separated by HPLC (high-performance liquid chromatography, for carotenoids) and LC-MS-MS (HPLC combined with mass spectrometry, for phenolic compounds) methods. As a result of the analytical studies, the following flavonoids had been identified: feruloyl glucopyranoside, quercetin rhamnopyranoside glucopyranoside, luteolin glucopyranoside arabinopyranoside, apigenin glucopyranoside arabinopyranoside, quercetin rhamnopyranoside, luteolin arabinopyranoside diglucopy-ranoside, hesperidine and luteolin glucuronide. According to the literature, the aglycones of these phenolic compounds exhibit MDR-reversal activity in vitro, and the connection between the phenolic content of Anastasia Black and MDR-reversal action was therefore studied by different analytical methods. The results of this study revealed that the identified flavonoids of Anastasia Black may be only partially responsible for the modulation of the MDR of mouse lymphoma cells. Other lipophilic compounds, most probably carotenoids, present in Russian black sweet pepper may act as inhibitors of MDR reversal.

Animals↗