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A new selection method for pepper transformation: callus-mediated shoot formation.

We used two genes, TMV-CP and PPI1 (pepper-PMMV interaction 1 transcription factor), to transform commercially important chili pepper (Capsicum annuum) inbred lines (P915, P409) by means of Agrobacterium co-culture. Eighteen independently transformed T0 plants were obtained. The most critical point in the pepper transformation protocol was the selection of shoots growing on calli--referred to as callus-mediated shoot formation (indirect shooting)--because shoots not grown from the callus (direct shooting from the wounded surface) developed into non-transformants. Selection of the correct right callus type also proved to be an important requirement for obtaining transformed peppers. Six different types of callus developed during the selection process. Shoots regenerated from two of these types, while one type regenerated significantly more shoots than the other types, suggesting that the capacity for shoot formation is callus type-specific. Although the transformation rate was low, transformation via callus-mediated shoot formation proved to be reproducible and was confirmed by Southern and Northern blot analyses. Based on the experimental data, we have succeeded in developing a new protocol for the selection and transformation of pepper and expect that it will be used in the future for pepper transformation.

Capsicum↗

Inhalation of vapor from black pepper extract reduces smoking withdrawal symptoms.

Previous studies have suggested that sensory cues associated with cigarette smoking can suppress certain smoking withdrawal symptoms, including craving for cigarettes. In this study we investigated the subjective effects of a cigarette substitute delivering a vapor of black pepper essential oil. Forty-eight cigarette smokers participated in a 3-h session conducted after overnight deprivation from smoking. Subjects were randomly assigned to one of three conditions: one group of smokers puffed on a device that delivered a vapor from essential oil of black pepper; a second group puffed on the device with a mint/menthol cartridge, and a third group used a device containing an empty cartridge. Subjects puffed and inhaled ad libitum from the device throughout the session during which no smoking was allowed. Reported craving for cigarettes was significantly reduced in the pepper condition relative to each of the two control conditions. In addition, negative affect and somatic symptoms of anxiety were alleviated in the pepper condition relative to the unflavored placebo. The intensity of sensations in the chest was also significantly higher for the pepper condition. These results support the view that respiratory tract sensations are important in alleviating smoking withdrawal symptoms. Cigarette substitutes delivering pepper constituents may prove useful in smoking cessation treatment.

Administration, Inhalation↗

The aflatoxin contamination of some pepper types sold in Turkey.

In the present study, 44 red-scaled pepper and 26 red powder pepper samples obtained from various spice retailers in Erzurum, and 20 isot (red pepper produced in Sanliurfa) were investigated for aflatoxin contamination using thin layer chromatography. Aflatoxin (B + G) was found in 8 red pepper samples (18.2%), and 3 red powder peppers (10.7%), and 1 isot sample (5%). The aflatoxin amount ranged from 1.1 to 97.5 ppb in all samples investigated. The highest amount of aflatoxin was found in red-scaled pepper. Species of Aspergillus, Penicillium and Rhizopus were generally contaminants of all samples.

Aflatoxins↗

Systematic identification pepper CaE2F transcription factor reveals the role of CaDPb in drought stress response.

The EARLY 2 FACTOR (E2F) transcription factor (TF) family plays a pivotal role in regulating plant development and adaptations to environmental stresses. However, the physiological function of E2Fs in pepper (Capsicum annuum L.) are not well elucidated. In this work, we conduct a comprehensive genome-wide annotation of the E2F family within the Zunla-1 pepper genome and further explore the biological roles of CaDPb in response to drought stress. Through systematic bioinformatics analysis, we identify a total of nine CaE2F genes within the Zunla-1 genome, categorizing them into three distinct subgroups. Additionally, we discover multiple cis-regulatory elements in the CaE2F promoter regions associated with responses to plant hormones and drought stress. Public RNA-seq datasets reveal distinct expression profiles of CaE2F genes across various pepper tissues and their responses to environmental stimuli and plant hormones. Subsequently, the CaDPb gene is further functionally verified in drought response. Our findings indicate that TRV2:CaDPb silenced pepper plants are more sensitivity to drought. Furthermore, we show that CaDPb participates in the regulation of reactive oxygen species (ROS) production, the expression of drought-responsive genes, and the modulation of stomatal aperture. Taken together, our findings provide a comprehensive characterization of E2F genes in pepper and offer insights into the biological function of CaDPb in pepper drought stress response.

Capsicum↗

Impact of occupational and inhalant allergy on rhinitis-specific quality of life in employees of bell pepper greenhouses in the Netherlands.

BACKGROUND: Rhinitis symptoms among bell pepper greenhouse employees can be caused by an allergy to occupational allergens, such as bell pepper pollen and predatory mites, and common inhalant allergens. OBJECTIVES: To estimate the effect of sensitization to these allergens on rhinitis-specific quality of life (QoL) during and outside the flowering period and to evaluate whether the QoL of sensitized employees is comparable with that of chrysanthemum greenhouse employees with rhinitis and an average population sample with perennial rhinitis. METHODS: We invited 233 employees with rhinitis symptoms to complete 2 rhinitis QoL questionnaires and to undergo skin prick testing with common inhalant allergens and occupational allergen extracts. Multiple regression analysis was used to estimate the impact of sensitization to the various allergens on QoL. RESULTS: Sensitization to bell pepper pollen had a significant negative effect on all the domain and mean QoL scores. The other allergens had no effect on QoL. A significant decrease in all the rhinitis scores was found outside the flowering period. There were no relevant differences in the mean scores of the different domains for both occupational groups. Greenhouse employees scored higher on limitations in activities and much lower on emotional, sleeping, and practical problems compared with individuals with perennial rhinitis. CONCLUSIONS: Bell pepper greenhouse employees are impaired in QoL because of their sensitization to bell pepper pollen, suggesting that bell pepper pollen is the most important occupational allergen in greenhouse workers with allergic symptoms. A common allergy does not have more impact on a person's day-to-day life than an occupational allergy; however, there is a clear difference in the way in which an occupational group is hampered compared with a nonoccupational group.

Adolescent↗

Characterization and quantitation of antioxidant constituents of sweet pepper (Capsicum annuum L.).

Sweet peppers (Capsicum annuum L.) cv. Vergasa have been studied at four maturity stages (immature green, green, immature red, and red). The individual phenolics (hydroxycinnamic acids and flavonoids), vitamin C (ascorbic acid and dehydroascorbic acid), and individual carotenoids were characterized and quantified. Five hydroxycinnamic derivatives and 23 flavonoids were characterized and quantified from the pericarp of sweet pepper by high-performance liquid chromatography-diode array detection-electrospray ionization-mass spectrometry. Identification was carried out by their UV spectra, chromatographic comparisons with authentic markers, identification of hydrolysis products, and tandem mass spectrometry analysis. Hydroxycinnamic derivatives, O-glycosides of quercetin, luteolin, and chrysoeriol, and a large number of C-glycosyl flavones have been characterized. Some of these compounds were found for the first time in nature. Clear differences in the individual and total phenolic content were detected between the different maturity stages. Immature green pepper had a very high phenolic content while green, immature red, and red ripe peppers showed a 4-5-fold reduction. Ascorbic acid was the main form of vitamin C, and its content increased as the pepper reached maturity. The red ripe stage had a relevant impact on the carotenoids content. Thus, immature green peppers showed the highest content of polyphenols, while red ripe fruits had the highest content of vitamin C and provitamin A.

Antioxidants↗

Induction of pepper cDNA encoding a lipid transfer protein during the resistance response to tobacco mosaic virus.

Pepper (Capsicum annuum) plants exhibit hypersensitive response (HR) against infection by many tobamoviruses. A clone encoding a putative nonspecific lipid transfer protein (CaLTP1) was isolated by differential screening of a cDNA library from resistant pepper leaves when inoculated with tobacco mosaic virus (TMV) pathotype P0. The predicted amino acid sequence of CaLTP1 is highly similar to that of the other plant LTPs. Southern blot analysis showed that a small gene family of LTP-related sequences was present in the pepper genome. Transcripts homologous to CaLTP1 accumulated abundantly in old leaves and flowers. CaLTP1 expression was induced in the incompatible interaction with TMV-P0 but was not induced in the compatible interaction with TMV-P1.2. In correlation with the temporal progression of HR in the inoculated leaves, CaLTP1 transcripts started to accumulate at 24 h after TMV-P0 inoculation, reaching a maximal level at 48 h. A strain of Xanthomonas campestris pv. vesicatoria (Xcv) that carries the bacterial avirulence gene, avrBs2, was infiltrated into leaves of a pepper cultivar containing the Bs2 resistance gene. A marked induction of CaLTP1 expression was observed in Xcv-infiltrated leaves. Effects of exogenously applied abiotic elicitors on CaLTP1 expression were also examined. Salicylic acid caused a rapid accumulation of CaLTP1 transcripts in pepper leaves and ethephon treatment also induced the expression of the CaLTP1 gene. Transient expression in the detached pepper leaves by biolistic gene bombardment indicated that CaLTP1 is localized mostly at the plant cell surface, possibly in the cell wall. These results suggest possible role(s) for LTPs in plant defense against pathogens including viruses.

Amino Acid Sequence↗

The potential use of a viral coat protein gene as a transgene screening marker and multiple virus resistance of pepper plants coexpressing coat proteins of cucumber mosaic virus and tomato mosaic virus.

Transgenic pepper plants coexpressing coat proteins (CPs) of cucumber mosaic virus (CMV-Kor) and tomato mosaic virus (ToMV) were produced by Agrobacterium-mediated transformation. To facilitate selection for positive transformants in transgenic peppers carrying an L gene, we developed a simple and effective screening procedure using hypersensitive response upon ToMV challenge inoculation. In this procedure, positive transformants could be clearly differentiated from the nontransformed plants. Transgenic pepper plants expressing the CP genes of both viruses were tested for resistance against CMV-Kor and pepper mild mottle virus (PMMV). In most transgenic plants, viral propagation was substantially retarded when compared to the nontransgenic plants. These experiments demonstrate that our transgenic pepper plants might be a useful marker system for the transgene screening and useful for classical breeding programs of developing virus resistant hot pepper plants.

Agrobacterium tumefaciens↗

Suppression of a ripening-related endo-1,4-beta-glucanase in transgenic pepper fruit does not prevent depolymerization of cell wall polysaccharides during ripening.

The function of the ripening-related endo-1,4-beta-D-glucanase (EGase) CaCel1 in fruit softening was investigated by suppression of CaCel1 gene expression in transgenic pepper (Capsicum annuum L.) plants using constitutive expression of a truncated sense CaCel1 transgene. In suppressed lines, immunodetectable CaCel1 protein and extractable CMCase activity were reduced to at or below the limit of detection in ripe mature red fruit, suggesting that in pepper ripening-related CMCase activity is the product of a single gene. However, the abundances of two mRNAs derived from the CaCel1 gene by differential transcription initiation were affected differently in suppressed lines. Accumulation of a 1.7 kb CaCel1 transcript was strongly suppressed, whereas the abundance of a 2.1 kb CaCel1 transcript was only partially reduced. This implies that the 1.7 kb mRNA is responsible for producing CaCel1 protein, while the 2.1 kb mRNA is translationally inactive, and as such is recalcitrant to co-suppression. Chelator-soluble polyuronides exhibited little or no depolymerization during ripening, but matrix glycans including xyloglucan were extensively depolymerized. Depolymerization of non-xyloglucan matrix glycans was the prominant cell wall change observed during pepper ripening. However, the lack of CaCel1 activity in suppressed fruit had no detectable effect on ripening-related matrix glycan depolymerization, which occurred at wild-type levels. Recombinant CaCel1 protein purified from a transgenic pepper line over-expressing functional CaCel1 was active against pepper matrix glycans in vitro, and showed greater activity against non-xyloglucan polysaccharides than against xyloglucan. Transgenic suppression of CaCel1 EGase activity has not identified the natural cell wall substrate for this enzyme, and shows that activities other than CaCel1 are responsible for the depolymerization of matrix glycans occurring during ripening in pepper.

Capsicum↗

Residues and half-lives of pyrethrins on field-grown pepper and tomato.

The dried flower heads of Tanacetum cinerariifolium Trev. (Family: Compositae) contain insecticidal compounds collectively called "pyrethrins." Pyrethrins are the subject of intense interest for use in crop protection because their toxicological properties permit control of certain insect species at application rates as low as 5-10 g AI acre(-1). Seedlings of sweet pepper, Capsicum annuum L. cv. Bell Boy Hybrid and tomato, Lycopersicon esculentum Mill. cv. Mountain Spring F1 Hybrid were planted and sprayed with a Multi-Purpose Insecticide formulation that contains 0.2% pyrethrins, 1.0% piperonyl butoxide (PBO), 88% diatomaceous earth, and 10.8% inert ingredients. The formulation was sprayed on pepper and tomato foliage when tomato fruits became red ripe and pepper became mature green at the rate of 6 lbs of formulated product per acre (5.4 and 27.2 g AI of pyrethrins and PBO, respectively). Following spraying, pepper and tomato leaves and fruits were collected at different time intervals for residue analysis using a high performance liquid chromatograph (HPLC) equipped with a UV detector. Residues of pyrethrins and PBO were generally higher on the leaves than fruits. Initial deposits (1 h following spraying) of pyrethrins were significantly higher on pepper than tomato fruits. Half-life (T1/2) values on pepper and tomato fruits did not exceed 2 h. Where concern exists over synthetic pesticide residues on treated crops and in the environment, pyrethrins are suitable alternatives that can be used to reduce the risk of exposure to synthetic pesticide residues.

Capsicum↗

Dietary red pepper ingestion increases carbohydrate oxidation at rest and during exercise in runners.

The effects of dietary hot red pepper on energy metabolism at rest and during exercise were examined in long distance male runners 18-23 yr of age. A standardized meal was given on the evening prior to the experiment. The subjects had a meal (2720 kJ) with or without 10 g of hot red pepper for breakfast. During rest (2.5 h after meal) and exercise (pedaling for 1 h at 150 W, about 60% VO2max, using cycling ergometry), expired gasses and venous blood were collected. The meal with hot red pepper significantly elevated respiratory quotient and blood lactate levels at rest and during exercise. Oxygen consumption at rest was slightly but nonsignificantly higher in the hot red pepper meal at 30 min after the meal. Plasma epinephrine and norepinephrine levels were significantly higher in those who had only hot red pepper at 30 min after the meal. These results suggest that hot red pepper ingestion stimulates carbohydrate oxidation at rest and during exercise.

Adolescent↗

A hot pepper cDNA encoding ascorbate peroxidase is induced during the incompatible interaction with virus and bacteria.

Capsicum annuum L. is infected by a number of viruses, including the tobacco mosaic virus (TMV). To study the defense-related genes that are induced by TMV in hot peppers, the pepper plant, which is susceptible to P1.2 but resistant to the P0 pathotype of TMV, was inoculated with TMV-P0. Differential screening isolated the genes that were specifically up- or down-regulated during the hypersensitive response (HR). The CaAPX1 cDNA clone that putatively encodes a polypeptide of cytosolic ascorbate peroxidase was selected as an up-regulated gene. It was isolated for further study. The full-length cDNA for CaAPX1, which is 972 bp long, contained the open-reading frame of 250-amino acid residues. A genomic Southern blot analysis showed that there were only limited copies of the CaAPX1 gene in the hot pepper genome. In hot pepper cv. Bugang, which is resistant to TMV-P0 and susceptible to TMV-P1.2, the CaAPX1 gene transcript was accumulated by TMV-P0, but not by TMV-P1.2 inoculation. CaAPX1 transcripts began to accumulate 24 h post-inoculation of TMV-P0, and increased gradually until 96 h. To investigate whether each transcript is induced by other stimuli, the plants were treated with various chemicals and wounding. A striking induction of the CaAPX1 transcript was observed at 2 h. It subsided 12 h after salicylic acid (SA), ethephon, and methyl jasmonate (MeJA) treatments. The response of the gene upon other pathogen infection was also examined by a bacterial pathogen (Xanthomonas campestris pv. vesicatoria race 3) inoculation. The CaAPX1 gene was induced in a hot pepper (C. annuum cv. ECW 20R) that was resistant to this bacterial pathogen, but not in a susceptible hot pepper (C. annuum cv. ECW). These results suggest the possible role(s) for the CaAPX1 gene in plant defense against viral and bacterial pathogen.

Amino Acid Sequence↗

Inheritance of resistance to Meloidogyne incognita race 2 in the hot pepper cultivar Carolina Cayenne (Capsicum annuum L.).

Root-knot nematodes of the genus Meloidogyne are important pathogens affecting vegetable crop production in Brazil and worldwide. The pepper species Capsicum annuum includes both hot and sweet peppers; very little emphasis has been placed on breeding sweet peppers for nematode resistance. We report on the inheritance of resistance to Meloidogyne incognita (Kofoid & White) Chitwood race 2 in the hot pepper cultivar Carolina Cayenne. The hot pepper cv. Carolina Cayenne was used as seed parent and the sweet pepper cv. Agronômico-8 was used as pollen parent to obtain the F(1) and F(2) generations and the backcross generations BC(11) and BC(12). The plants were inoculated with M. incognita race 2 at a rate of 60 eggs/ml of substrate and, after a suitable incubation period, the numbers of root galls and egg masses per root system were evaluated on each plant. Broad- (0.77 and 0.72) and narrow-sense (0.77 and 0.63) heritability estimates were high for both root galls and egg masses, respectively. The mean degree of dominance was estimated as 0.29 and 0.25 for numbers of galls and egg masses, respectively; these estimates were not significantly different from 0, indicating a predominantly additive gene action. The results were consistent with a hypothesis of monogenic resistance in Carolina Cayenne.

Animals↗

Molecular cloning of a pepper gene that is homologous to SELF-PRUNING.

"Determinate" and "indeterminate" inflorescences in plants are controlled by a single recessive gene, for example, SELF-PRUNING (SP) in Solanum lycopersicum, TERMINAL FLOWER1 in Arabidopsis, CENTRORADI-ALIS in Antirrhinum, and CENTRORADIALIS-like gene in tobacco. Pepper (Capsicum annuum L.) is an indeterminate species in which shoots grow indefinitely. In this study, we cloned and characterized the pepper SP-like gene (CaSP). RT-PCR revealed that the CaSP transcript accumulates to higher levels in floral buds than in other organs. Comparison of genomic DNA and cDNA sequences from indeterminate and determinate pepper plants revealed the insertion of a single base in the first exon of CaSP in the determinate pepper plants. CaSP is annotated in linkage group 8 (chromosome 6) of the SNU2 pepper genetic map and showed similar synteny to SP in tomato. Transgenic tobacco plants overexpressing CaSP displayed late-flowering phenotypes similar to the phenotypes caused by overexpression of CaSP orthologs in other plants. Collectively, these results suggest that pepper CaSP is an ortholog of SP in tomato.

Amino Acid Sequence↗

Screening of aflatoxins in Shiro and ground red pepper in Addis Ababa.

Aflatoxin contamination of Shiro and ground red pepper samples collected from government owned good stores, retail shops and open markets in. Addis Ababa was investigated. From sixty samples each of ground red pepper and Shiro, 8 (13.33%) and 5 (8.33%) were positive for aflatoxins, respectively. Only aflatoxin B1 was detected in both types of foodstuff. There was no significant difference between the proportion of aflatoxin contained in both ground red pepper and Shiro samples. Aflatoxin levels in Shiro and ground red pepper positive samples ranged from 100 to 500 ppb and 250 to 525 ppb, respectively. The mean levels of contamination by aflatoxin in ground red pepper was significantly higher (p < 0.05) than that of Shiro. Though statistically not significant, the number of positive samples were more in samples from open markets when compared to samples from government owned food stores and retail shops. Conclusions are drawn that ground red pepper and shiro traded in Addis Ababa can be considered as high risk commodities for which routine survey of aflatoxins may be necessary.

Aflatoxins↗

Genome-wide identification of CHY zinc finger and RING finger (CHYR) genes in pepper and functional characterization of CaCHYR5 in response to Phytophthora capsici infection.

CHY zinc finger and RING finger (CHYR) proteins play crucial roles in the growth and development, as well as stress response. To date, no systematic or comprehensive analysis of the CHYR gene family has been performed in pepper (Capsicum annuum L.). In this study, we identified 8 CaCHYR genes (CaCHYR1-CaCHYR8), which were classified into 3 groups based on phylogenetic relationships. CaCHYR members within the same group exhibited similar distributions of conserved motifs and exon-intron structures. Chromosomal localization analysis showed that 8 CaCHYR genes were unevenly distributed on 6 chromosomes. Segmental duplication, rather than tandem duplication, was found to be the major contributor to the expansion of this gene family. CaCHYR genes feature a variety of cis-elements involved in developmental processes, phytohormone responses, and stress adaptation. Expression analysis based on RNA-seq data revealed that CaCHYR genes exhibited distinct spatial expression patterns across different tissues and in response to Phytophthora capsici infection (PCI), and quantitative real-time PCR (qRT-PCR) further confirmed that three of them (CaCHYR2, CaCHYR3, and CaCHYR5) exhibited altered expression under PCI. Furthermore, transient overexpression of CaCHYR5 in pepper leaves increased susceptibility to PCI, suggesting its potential negative regulatory role in pepper defense against P. capsici. Collectively, these findings reveal the expression patterns and regulatory functions of pepper CHYR genes in growth and development, laying a groundwork for breeding pepper cultivars tolerant to PCI.

Phytophthora capsici infection (PCI)↗

The organization of mitochondrial atp6 gene region in male fertile and CMS lines of pepper (Capsicum annuum L.).

The mitochondrial atp6 gene in male fertile (N) and CMS (S) pepper has previously been compared and was found to be present in two copies (Kim et al. in J Kor Soc Hort Sci 42:121-127 2001). In the current study, these atp6 copies were amplified by an inverse PCR technique, and the coding region as well as the 5' and 3' flanking regions were sequenced. The atp6 copies in CMS pepper were detected as one intact gene and one pseudogene, truncated at the 3' coding region. When the atp6 genes in pepper were compared to other plant species, pepper, potato, and petunia all possessed a sequence of 12 identical amino acids at the 3' extended region, which was considered a hallmark of the Solanaceae family. Northern blot analysis showed differences in mRNA band patterns between CMS and restorer lines, indicating that atp6 gene is one of the candidates for CMS in pepper.

Amino Acid Sequence↗

EST and microarray analyses of pathogen-responsive genes in hot pepper (Capsicum annuum L.) non-host resistance against soybean pustule pathogen (Xanthomonas axonopodis pv. glycines).

Large-scale single-pass sequencing of cDNA libraries and microarray analysis have proven to be useful tools for discovering new genes and studying gene expression. As a first step in elucidating the defense mechanisms in hot pepper plants, a total of 8,525 expressed sequence tags (ESTs) were generated and analyzed in silico. The cDNA microarray analysis identified 613 hot pepper genes that were transcriptionally responsive to the non-host soybean pustule pathogen Xanthomonas axonopodis pv. glycines ( Xag). Several functional types of genes, including those involved in cell wall modification/biosynthesis, transport, signaling pathways and divergent defense reactions, were induced at the early stage of Xag infiltration. In contrast, genes encoding proteins that are involved in photosynthesis, carbohydrate metabolism and the synthesis of chloroplast biogenetic proteins were down-regulated at the late stage of Xag infiltration. These expression profiles share common features with the expression profiles elicited by other stresses, such as fungal challenge, wounding, cold, drought and high salinity. However, we also identified several novel transcription factors that may be specifically involved in the defense reaction of the hot pepper. We also found that the defense reaction of the hot pepper may involve the deactivation of gibberellin. Furthermore, many genes encoding proteins with unknown function were identified. Functional analysis of these genes may broaden our understanding of non-host resistance. This study is the first report of large-scale sequencing and non-host defense transcriptome analysis of the hot pepper plant species. (The sequence data in this paper have been submitted to the dbEST and GenBank database under the codes 10227604-10236595 and BM059564-BM068555, respectively. Additional information is available at http://plant.pdrc.re.kr/ks200201/pepper.html).

Capsicum↗