[Determination of active principles of capsicum].
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Various concentrations of chilli extract were tested in 2 stages on the growth of a human buccal mucosa fibroblast cell line. Firstly, concentrations of 25, 75, 150 and 300 micrograms/ml were tested on the cell line for 6 days followed by a repeat which included concentrations of 400 and 500 micrograms/ml. In the latter, cell growth was monitored for 18 days. From day 3 (72 hr) suppression of cell growth was evident with concentrations 300-500 micrograms/ml. Total cell death occurred at 16 days with 300 micrograms/ml and at 6 days with 400-500 micrograms/ml. With the lower concentrations, 25-150 micrograms/ml, the daily counts were lower than the control but the difference was not statistically significant. Growth continued unabated. It is therefore concluded that cytopathic effect of chilli extract to fibroblasts is concentration dependent.
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We have determined the nucleotide sequence of the plastid psbL gene from bell pepper. This gene has an ACG as a first codon. Isolation of RNA from pepper leaves and ripe fruits and subsequent sequencing of the psbL cDNA revealed that this ACG codon is post-transcriptionally edited into an AUG initiation codon in both leaves and fruits. These data indicate that the RNA editing machinery which exists in chloroplasts is still functional in chromoplasts from ripe fruits.
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A new, highly sensitive method for quantitative separation and determination of capsaicin and dihydrocapsaicin by high-performance liquid chromatography with electrochemical detection is elaborated. The method employs a Cosmosil 5Ph column eluted with 100 mM potassium dihydrogenphosphate containing 45% acetonitrile (pH 5.0) at a flow-rate of 1.0 ml/min, and an electrochemical detector (at a potential of +750 mV versus the Ag/AgCl electrode). The detection limits for both capsaicin and dihydrocapsaicin were 12 pg (39 fmol) at a signal-to-noise ratio of 3:1. By applying this method, the biological half-life of capsaicin in the rat was investigated.
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Northern blot and in situ hybridization analyses revealed that a pepper basic class II chitinase gene (CAChi2) is constitutively expressed in floral organs and root endodermis, but not in leaf, stem and fruit of pepper. Resistance of pepper leaves to Colletotrichum coccodes infection at a late growth stage was correlated with induction of beta-1,3-glucanase and PR-1 mRNA, but not of chitinase (CAChi2) mRNA. Transcriptional activation of the CAChi2 gene in pepper leaves occurred during anthracnose development. The CAChi2 transcripts were mainly localized in phloem cells of vascular tissues of pepper leaves infected with C. coccodes. The CAChi2 gene was also differentially induced in leaf and stem tissue by treatment with abscisic acid (ABA), sodium chloride or drought. Strong accumulation of the CAChi2 transcripts occurred in pepper stem tissues due to high salt and drought, and also due to treatment with ABA. These results suggest involvement of the chitinase gene in protection of pepper plants against the pathogen, but also document cross talk with stress signals mediated by ABA, high salinity and drought.