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Carotenoid biosynthesis changes in five red pepper (Capsicum annuum L.) cultivars during ripening. Cultivar selection for breeding.

Changes in the biosynthesis of individual carotenoid pigments have been investigated during fruit ripening of five cultivars of red pepper (Capsicum annuum L.): Mana, Numex, Belrubi, Delfin, and Negral (a chlorophyll-retaining mutant when ripe). The study was carried out throughout the ripening process, and with special emphasis on the ripe stage, to discover possible differences between cultivars and to characterize these by their carotenoid pattern and content for selecting the best varieties for breeding programs. Ripening fruit of the five cultivars showed the typical and characteristic pattern of carotenoid biosynthesis for the Capsicum genus. In the five cultivars, lutein and neoxanthin, both characteristic chloroplast pigments, decreased in concentration with ripening and eventually disappeared. beta-Carotene, antheraxanthin, and violaxanthin increased in concentration, and other pigments were biosynthesized de novo: zeaxanthin, beta-cryptoxanthin, capsanthin, capsorubin, capsanthin-5,6-epoxide, and cucurbitaxanthin A. A pool of zeaxanthin stands out of the rest of pigment during ripening, which reveals the importance of this pigment as a branching point in the carotenoid biosynthesis in Capsicum. Quantitatively, Negral cultivar showed the highest increase in total carotenoid content (48. 39-fold), followed by Mana and Delfin with 38.03- and 36.8-fold, respectively, and by Belrubi and Numex with 28.03- and 23.48-fold, respectively. In all the red varieties, there was an inverse relationship between total carotenoid content and the red to yellow isochromic pigment fraction ratio (R/Y) and the capsanthin-to-zeaxanthin ratio (Caps/Zeax). This seems to be related to the carotenogenic capacity of the cultivar, and thus selection and breeding should not only seek a higher total carotenoid content but also attempt to increase these ratios. In the present study, the cultivar Mana had the highest total carotenoid content (13 208 mg/kg dwt), but the lowest R/Y (1.25) and Caps/Zeax (3.38) ratios, which are therefore the parameters to improve. The cultivar Negral had a high carotenoid content (8797 mg/kg dwt) and high R/Y and Caps/Zeax ratios and could be used for transfer of these characters in direct crosses with the cultivar Mana. The cultivar Numex had the highest Caps/Zeax ratio (7.17) and is thus an ideal progenitor for this character.

Capsicum↗

Formation of volatile compounds during heating of spice paprika (Capsicum annuum) powder.

Spice paprika (red pepper; Capsicum annuum) is the most cultivated spice worldwide and is used mainly for its color and pungency. However, current research is also focusing on the flavor as an important parameter. This paper deals with the kinetics of the formation of those volatiles that indicate a decrease in spice paprika quality due to Maillard reaction, hydrolytic reactions, and oxidative degradation reactions of lipids such as fatty acids and carotenoids. Spice paprika volatiles were quantitatively analyzed by means of headspace gas chromatography (HS-GC) and solid-phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS). The kinetics of their formation were investigated, and the respective activation energies determined. Strecker aldehyde, acetone, and methanol formation followed a pseudo-zero-order reaction kinetic, and formation of dimethyl sulfide (DMS) was characterized by a first-order kinetic. The activation energies determined were between 86.3 and 101.8 for the Strecker aldehydes acetaldehyde (AA), 2-methylpropanal (2-MP), 3-methylbutanal (3-MB), and 2-methylbutanal (2-MB), 130.7 for acetone, 114.2 for methanol, and 109.7 kJ/mol for DMS. The amounts of Strecker aldehydes formed were correlated to the concentrations of the corresponding free amino acids present in the samples. The formation of hexanal and 6-methyl-5-hepten-2-one in Capsicum annuum during processing was confirmed, and the formation of beta-ionone was probably described for the first time. During heating, the concentration of hexanal increased rapidly. The formation of 6-methyl-5-hepten-2-one confirms that Capsicum annuum fruits contain lycopene.

Capsicum↗

Screening Capsicum accessions for capsaicinoids content.

Ninety Capsicum accessions selected from the USDA Capsicum germplasm collection were screened for their capsaicinoids content using gas hromatography with nitrogen phosphorus detection (GC/NPD). Fresh fruits of Capsicum chinense, C. frutescens, C. baccatum, C. annuum, and C. pubescens were extracted with methanol and analyzed for capsaicin, dihydrocapsaicin, and nordihydrocapsaicin. Mass spectrometry of the fruit crude extracts indicated that the molecular ions at m/z 305, 307, and 293, which correspond to capsaicin, dihydrocapsaicin, and nordihydrocapsaicin, respectively, have a common benzyl cation fragment at m/z 137 that can be used for monitoring capsaicinoids in pepper fruit extracts. Capsaicin and dihydrocapsaicin were the dominant capsaicinoids detected. Capsaicin concentrations were typically greater than dihydrocapsaicin. Concentrations of total capsaicinoids varied from not detectable to 11.2 mg fruit(-1). Statistical analysis revealed that accession PI-441624 (C. chinense) had the highest capsaicin content (2.9 mg g(-1) fresh fruit) and accession PI-497984 (C. frutescens) had the highest dihydrocapsaicin content (2.3 mg g(-1) fresh fruit). Genebank accessions PI-439522 (C. frutescens) and PI-497984 contained the highest concentrations of total capsaicinoids.

Capsaicin↗

Improvement of shelf-life and microbiological quality of minimally processed refrigerated capsicum by gamma irradiation.

Shelf-life, microbiological and chemical quality of minimally processed capsicum subjected to radiation doses of 1, 2 and 3 kGy followed by storage at 5 degrees C and 10 degrees C were evaluated. Irradiation at an optimal dose of 2 kGy reduced the initial bacterial population by 2-3 log cycles and eliminated the coliforms Listeria and Yersinia. Chemical analysis revealed that the initial contents of ascorbic acid (127.7 mg/100 g), carotenoid (110 microg/100 g) and chlorophyll (7.75 mg/g) were reduced marginally by 5-10% with increasing radiation dose. However, during subsequent storage, up to 4 weeks, the temperature-dependent losses in vitamin C and chlorophyll content of irradiated samples were less compared with non-irradiated samples. The total carotenoid content of capsicum irradiated and stored for up to 2 weeks at 10 degrees C showed a similar trend. Thus, gamma irradiation at 2 kGy was found to improve both the hygienic quality and shelf-life without affecting the nutritional quality of minimally processed capsicum.

Ascorbic Acid↗

Capsicum production, technology, chemistry, and quality. Part 1: History, botany, cultivation, and primary processing.

The genus Capsicum (Fam. Solanaceae) was known to ancient cultures and was more recently historically associated with the discovery of the New World. This genus provides many species and varieties used in flavoring foods popular in the cuisines of many parts of the world. From the pungent chilli to the colorful paprika and the bell pepper, with its remarkable aroma, the genus is of great interest for its chemistry, sensory attributes, and physiological action. The Capsicums, among the spices, are second only to black pepper in trade both in volume and value. The production of the different pungency forms, the processed seasonings, and the concentrated oleoresins, through technologically advanced processes and in specified standard grades, are critically reviewed. The pungency of Capsicum fruits, its evaluation, chemical structure relationship, its increasing acceptance and preference by a variety of populations are of great research interest. The wide traditional use in the growing regions and its intense physiological effects have attracted the attention of researchers of many different disciplines. These aspects are reviewed in four sequential parts. Part I deals with history, botany, cultivation, and primary processing.

Agriculture↗

Capsicum--production, technology, chemistry, and quality. Part IV. Evaluation of quality.

Capsicum fruits are popular worldwide and are used in the cuisines of both the developing and the developed countries. With its different varieties, forms, and uses, the spice capsicum contributes to the entire gamut of sensory experience--color as finely ground paprika powder or extract in sausages, goulash, cheese, and snacks; both pungency and color as the many varieties of chillies used in Mexican, African, Indian, and southeast Asian cuisines; color, aroma, and mild pungency as the fresh green chillies used in many of the growing countries; and appearance, color, aroma, and texture as fresh fruit in salads and as a pickled and canned product. In three earlier parts in this series, the varieties, cultivation, and primary processing; the processed products, world production, and trade; and the chemistry of the color, aroma, and pungency stimuli have been reviewed. In this part, the evaluation of quality through instrumental determination of the causal components and the sensory evaluation of color, aroma, and pungency are discussed. Several methods for quantitative determination of the stimuli and the sensory evaluation of the responses to the stimuli are reviewed. The problems of sensory evaluation of color, aroma, and pungency, the dominant attributes for validation of the instrumentally determined values for carotenoids, volatiles, or particular fractions, and total and individual capsaicinoids are specifically discussed. Summarized details of selected instrumental methods for evaluating the stimuli, which are either validated by correlation to sensorily perceived responses or to adopted standards, are given along with representative data obtained for discussing the adequacy and reliability of the methods. Pungency as a specific gustatory perception and the many methods proposed to evaluate this quality are discussed. A recommended objective procedure for obtaining reproducible values is discussed, and a method for relating different panel results is shown. With such a method, highly significant correlations have been shown between estimated total capsaicinoids and the determined pungency. The estimation of total capsaicinoids by any simple, reliable method is shown to be adequate for quality control of pungency of Capsicum fruits.

Capsicum↗

The relationship between high fibrinolytic activity and daily capsicum ingestion in Thais.

Capsicum, a hot appetizer and seasoning, has been found to induce increased fibrinolytic activity and simultaneously cause hypocoagulability of blood when ingested or when retained in the mouth for a short time. The effect on fibrinolysis and blood coagulation of capsicum can be reproduced in the same subjects within a short time after the first stimulation. More investigations on this effect may lead to the discovery of some ideal drugs for both treatment and prevention of thromboembolism. Fibrinolytic activity measured by euglobulin lysis time in 88 Thai subjects (mean +/- SD = 167 +/- 66.84 min) was significantly higher than in 55 American whites (mean +/- SD = 254 +/- 126.70 min) residing in Thailand for a period of time (p less than 0.001). The Thai people consume capsicum with their meals. Their fibrinolytic activity, therefore, is activated several times during the day and this activation could be an important factor in causing high fibrinolytic activity. This customary habit of food ingestion is very likely a factor contributing to the racial difference in fibrinolysis. Furthermore, the Thais also have lower plasma fibrinogen and higher antithrombin III compared to Americans. These could certainly be additional factors, in addition to fibrinolytic activity, that play a role in the rarity of thromboembolism among Thais.

Adolescent↗

The coat protein is required for the elicitation of the Capsicum L2 gene-mediated resistance against the tobamoviruses.

In Capsicum, the resistance against tobamoviruses conferred by the L2 gene is effective against all but one of the known tobamoviruses. Pepper mild mottle virus (PMMoV) is the only virus which escapes its action. To identify the viral factors affecting induction of the hypersensitive reaction (HR) mediated by the Capsicum spp. L2 resistance gene, we have constructed chimeric viral genomes between paprika mild mottle virus (PaMMV) (a virus able to induce the HR) and PMMoV. A hybrid virus with the PaMMV coat protein gene substituted in the PMMoV-S sequences was able to elicit the HR in Capsicum frutescens (L2L2) plants. These data indicate that the sequences that affect induction of the HR mediated by the L2 resistance gene reside in the coat protein gene. Furthermore, a mutant that codes for a truncated coat protein was able to systemically spread in these plants. Thus, the elicitation of the host response requires the coat protein and not the RNA.

Amino Acid Sequence↗

Effects of alfalfa extract, anise, capsicum, and a mixture of cinnamaldehyde and eugenol on ruminal fermentation and protein degradation in beef heifers fed a high-concentrate diet.

Four Holstein heifers (360 +/- 22 and 450 +/- 28 kg of BW in Exp. 1 and 2, respectively) fitted with ruminal trocars were used in 4 x 4 Latin square designs to evaluate the effects on ruminal microbial fermentation of the following: Exp. 1, no additive, alfalfa extract (30 g/d, AEX), a mixture of cinnamaldehyde (0.18 g/d) and eugenol (0.09 g/d; CIE1), and AEX and CIE1 in combination; and Exp. 2, no additive, anise oil (2 g/d), capsicum oil (1 g/d), and a mixture of cinnamaldehyde (0.6 g/d) and eugenol (0.3 g/d). Heifers were fed a 90:10 concentrate:barley straw diet (16% CP; 25% NDF) for ad libitum intake. Each period consisted of 15 d for adaptation and 6 d for sampling. On d 16 to 18, DM and water intakes were measured. On d 19 to 21 ruminal contents were sampled at 0, 3, 6, 9, and 12 h after feeding to determine ruminal pH and the concentrations of VFA, L-lactate, large peptides, small peptides plus AA (SPep+AA), and ammonia N. On d 20 and 21, samples of ruminal fluid were collected at 0 and 3 h after feeding to determine protozoal counts. In Exp. 1, CIE1 and AEX decreased (P < 0.05) total DMI, concentrate DMI, and water intake. The increase (P < 0.05) in SPep+AA and the decrease (P < 0.05) in ammonia N when supplementing CIE1 suggest that deamination was inhibited. Treatment AEX increased (P < 0.05) the acetate to propionate ratio, which is less efficient for beef production. Treatment CIE1 increased (P < 0.05) counts of holotrichs. Effects of AEX and CIE1 were not additive for many of the measured metabolites. In Exp. 2, treatments had no effect on ruminal pH, total VFA concentration, and butyrate proportion. The capsicum oil treatment increased (P < 0.05) DMI, water intake, and SPep+AA N concentration and decreased (P < 0.05) acetate proportion, branched-chain VFA concentration, and large peptide N concentration. The cinnamaldehyde (0.6 g/d) and eugenol (0.3 g/d) treatment decreased (P < 0.05) water intake, acetate proportion, branched-chain VFA, L-lactate, and ammonia N concentrations and increased (P < 0.05) propionate proportion and SPep+AA N concentration. The anise oil treatment decreased (P < 0.05) acetate to propionate ratio, branched-chain VFA and ammonia N concentrations, and protozoal counts. The results indicate that at the doses used a mixture of cinnamaldehyde and eugenol, anise oil, and capsicum oil may be useful as modifiers of rumen fermentation in beef production systems.

Acrolein↗

Effects of red chilli (Capsicum frutescens L) on rats.

The effects of red chilli were examined in 12-w-old male Wistar rats fed a commercial diet mixed with ground Capsicum frutescens L fruits at levels of 2% and 10% (w/w) for 8 w. When compared with controls, the general health, body weight gain, feed intake and feed efficiency were not adversely affected on the rats on feed with 2% Capsicum, while the feed intake and growth rate were depressed and exfoliation of the intestinal epithelium into the lumen and cytoplasmic fatty vacuolation and necrosis of the centrilobular hepatocytes occurred in rats at 4 and 8 w on feed with 10% Capsicum. This correlated with changes in hematology, serum enzyme profiles and other serum constituents.

Animals↗

An evaluation of the relative potential public health concern for the self-defense spray active ingredients oleoresin capsicum, o-chlorobenzylidene malononitrile, and 2-chloroacetophenone.

In 1996, the New York State Department of Health was charged by the State Legislature to develop regulations regarding the types of self-defense spray devices which could lawfully be purchased, possessed, and used in New York State. Prior to this legislation, sale or possession of self-defense spray devices in New York State was illegal. The Department of Health used existing data to evaluate three commonly used self-defense spray active ingredients (oleoresin capsicum, o-chlorobenzylidene malononitrile, and 2-chloroacetophenone) with respect to their relative toxicity and their involvement in accidental poisonings. Based on the balance of the available information, the Department of Health determined that oleoresin capsicum posed a lower public health concern than o-chlorobenzylidene malononitrile or 2-chloroacetophenone, and developed a rule that specifies oleoresin capsicum as the only active ingredient to be used in self-defense sprays for sale and use in New York State.

Accidents↗

Purification and characterization of farnesyl pyrophosphate synthase from Capsicum annuum.

Farnesyl pyrophosphate synthase (FPP) displaying dimethylallyl transferase activity (EC 2.5.1.1) and geranyl transferase activity (EC 2.5.1.10) was purified from Capsicum fruits. This prenyltransferase has a molecular mass of 89,000 +/- 5000 Da resulting from the association of two apparently identical subunits having a molecular mass of 43,000 +/- 2000 Da. Antibodies raised against Capsicum FPP synthase selectively blocked the transferase activity. Analysis of the immunological relationships between FPP synthase and geranylgeranyl pyrophosphate synthase (EC 2.5.1.1, EC 2.5.1.10 and EC 2.5.1.30) revealed that these two enzymes though performing the same mechanism of catalysis and accepting identical substrates have different antigenic determinants. Thus, in connection to previous work, this immunological study suggests that Capsicum FPP is strictly located in the extraplastidial compartment.

Cations, Divalent↗

Isolation and purification of the hypoglycaemic principle present in Capsicum frutescens.

Capsicum frutescens has been used to treat diabetes mellitus by traditional healers in Jamaica. Purification experiments employing thin layer chromatography (TLC) and high performance liquid chromatography led to the extraction of the active principle, capsaicin. Purified capsaicin caused a decrease in blood glucose levels to 4.91 +/- 0.52 (n = 6) mmol/dL versus 6.40 +/- 0.13 mmol/dL (n = 6) for the control (p < 0.05) at 2.5 h in an OGTT in dogs. There was a concomitant elevation in plasma insulin levels (p < 0.05). In conclusion, it can be stated that capsaicin is the major constituent of Capsicum frutescens that is responsible for the hypoglycaemic episodes seen in the dogs. It is also apparent that the latter is mediated by insulin release.

Animals↗

Molecular mapping of capsaicinoid biosynthesis genes and quantitative trait loci analysis for capsaicinoid content in Capsicum.

Quantitative variation in the accumulation of two major capsaicinoids responsible for pungency in the fruit of chile peppers, capsaicin and dihydrocapsaicin, was analyzed in a cross between the non-pungent Capsicum annuum parent cv. Maor and a pungent Capsicum frutescens parent, accession BG 2816. In order to identify quantitative trait loci (QTLs) for capsaicinoid content, we employed the bulked segregant analysis method and screened bulked DNA from F2 individuals at the extremes of the distribution of capsaicinoid content with RAPD primers. Screening with 400 primers allowed the identification of three loci that were polymorphic between the bulks. These RAPD markers were converted to SCARs and subsequently mapped with additional RFLP markers to chromosome 7 of pepper. QTL interval analysis for individual and total capsaicinoid content identified a major QTL, termed cap, which explained 34-38% of the phenotypic variation for this trait in two growing environments. For all measurements, the allele of the pungent parent BG 2816 at cap contributed to the increased level of pungency. To determine whether known structural genes in the pathway could define a candidate for this QTL, 12 clones obtained from differentially expressed transcripts from placental tissue in pungent peppers were also mapped. None of them had a significant effect on this trait, nor did the allelic state at the locus C, the on/off switch for pungency in pepper, located on chromosome 2. The identity of cap and its effect on capsaicin content in other backgrounds will be addressed in future studies.

Alleles↗

QTL mapping of anthracnose ( Colletotrichum spp.) resistance in a cross between Capsicum annuum and C. chinense.

Anthracnose fruit rot is an economically important disease that affects pepper production in Indonesia. Strong resistance to two causal pathogens, Colletotrichum gloeosporioides and C. capsici, was found in an accession of Capsicum chinense. The inheritance of this resistance was studied in an F(2) population derived from a cross of this accession with an Indonesian hot pepper variety ( Capsicum annuum) using a quantitative trait locus (QTL) mapping approach. In laboratory tests where ripe fruits were artificially inoculated with either C. gloeosporioides or C. capsici, three resistance-related traits were scored: the infection frequency, the true lesion diameter (averaged over all lesions that actually developed), and the overall lesion diameter (averaged over all inoculation points, including those that did not develop lesions). One main QTL was identified with highly significant and large effects on all three traits after inoculation with C. gloeosporioides and on true lesion diameter after inoculation with C. capsici. Three other QTL with smaller effects were found for overall lesion diameter and true lesion diameter after inoculation with C. gloeosporioides, two of which also had an effect on infection frequency. Interestingly, the resistant parent carried a susceptible allele for a QTL for all three traits that was closely linked to the main QTL. The results with C. capsici were based on less observations and therefore less informative. Although the main QTL was shown to have an effect on true lesion diameter after inoculation with C. capsici, no significant QTL were identified for overall lesion diameter or infection frequency.

Ascomycota↗

QTL analysis for capsaicinoid content in Capsicum.

Pungency or "heat" found in Capsicum fruit results from the biosynthesis and accumulation of alkaloid compounds known as capsaicinoids in the dissepiment, placental tissue adjacent to the seeds. Pepper cultivars differ with respect to their level of pungency because of quantitative and qualitative variation in capsaicinoid content. We analyzed the segregation of three capsaicinoids: capsaicin, dihydrocapsaicin and nordihydrocapsaicin in an inter-specific cross between a mildly pungent Capsicum annuum 'NuMex RNaky' and the wild, highly pungent C. frutescens accession BG2814-6. F(3) families were analyzed in three trials in California and in Israel and a dense molecular map was constructed comprised mostly of loci defined by simple sequence repeat (SSR) markers. Six QTL controlling capsaicinoid content were detected on three chromosomes. One gene from the capsaicinoid biosynthetic pathway, BCAT, and one random fruit EST, 3A2, co-localized with QTL detected in this study on chromosomes 3 and 4. Because one confounding factor in quantitative determination of capsaicinoid is fruit size, fruit weight measurements were taken in two trials. Two QTL controlling fruit weight were detected, however, they did not co-localize with QTL detected for capsaicinoid content. The major contribution to the phenotypic variation of capsaicinoid content (24-42% of the total variation) was attributed to a digenic interaction between a main-effect QTL, cap7.1, and a marker located on chromosome 2 that did not have a main effect on the trait. A second QTL, cap7.2 is likely to correspond to the QTL, cap, identified in a previous study as having pronounced influence on capsaicinoid content.

Capsaicin↗

Induction of a cytosolic pyruvate kinase 1 gene during the resistance response to Tobacco mosaic virus in Capsicum annuum.

Hot pepper (Capsicum annuum L. cv. Bugang) plants exhibit a hypersensitive response (HR) upon infection by Tobacco mosaic virus (TMV) pathotype P(0). Previously, to elucidate molecular mechanism that underlies this resistance, hot pepper cv. Bugang leaves were inoculated with TMV-P(0) and genes specifically up-regulated during the HR were isolated by microarray analysis. One of the clones, Capsicum annuum cytosolic pyruvate kinase 1 (CaPK(c)1) gene was increased specifically in the incompatible interaction with TMV-P(0). The expression of CaPK(c)1 gene was also triggered not only by various hormones such as salicylic acid (SA), ethylene, and methyl jasmonate (MeJA), but also NaCl and wounding. These results suggest that CaPK(c)1 responds to several defense-related abiotic stresses in addition to TMV infection.

Amino Acid Sequence↗

Isolation and characterization of a lipid transfer protein expressed in ripening fruit of Capsicum chinense.

A novel LTP (CcLTP) from a Capsicum chinense cv Habanero was isolated from a fruit-specific SSH library. While this gene shares similarity with other LTPs, it is considerably larger than any lipid transfer protein reported to date and has a neutral predicted pI. CcLTP is consistently expressed in seedlings from three Capsicum species. It is also present at very high levels in ripening and mature fruit in C. chinense, but not in fruit of any C. annuum or C. frutescens varieties examined. We have obtained 3.8 kb of sequence containing the CcLTP gene and isolated two forms of mRNA transcripts which result from an alternative splicing event. Both transcripts are full-length cDNAs with putative open reading frames of 492 bp and 519 bp, encoding proteins of 164 and 173 amino acids, respectively, which differ only by an insertion of 9 amino acids. Both splice variants are detected consistently via RT-PCR. A 19 bp deletion in the promoter region differentiates C. chinense CcLTP from that of C. annuum and C. frutescens. The protein and its expression are characterized in C. chinense fruit, and a possible role in pepper fruit ripening and maturation is discussed.

Alternative Splicing↗