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Ornithine decarboxylase gene (CaODC1) is specifically induced during TMV-mediated but salicylate-independent resistant response in hot pepper.

A gene encoding putative ornithine decarboxylase (ODC) has been isolated by differential screening of a cDNA library from the resistant hot pepper (Capsicum annuum L.) inoculated with avirulent tobacco mosaic virus (TMV) pathotype P0. In hot pepper plants, transcripts of the CaODC1 (C. annuum ODC1) gene started to accumulate at 24 h post-inoculation of TMV-P0 and the signal was spread systemically. The transcript level of CaODC1 was increased rapidly in a hot pepper resistant to Xanthomonas campestris pv. vesicatoria (Xcv) but not in a susceptible hot pepper after inoculation. These results suggest possible role(s) for CaODC1 in plant defense against a broad range of pathogens including viruses and bacteria.

Amino Acid Sequence↗

Isolation, partial sequencing, and expression of pathogenesis-related cDNA genes from pepper leaves infected by Xanthomonas campestris pv. vesicatoria.

Specific cDNAs showing differential expression in bacteria-infected pepper leaves as opposed to healthy leaves were isolated from a pepper cDNA library from hypersensitive response (HR) lesions of leaves infected with an avirulent strain of Xanthomonas campestris pv. vesicatoria. Among a total of 282 cDNA clones tested, 36 individual cDNA genes (13%) hybridized strongly or differentially to the cDNA probes from bacteria-infected leaves. Ten Capsicum Annuum-Induced (CAI) genes encoding putative thionin, lipid transfer protein I and II, osmotin (PR-5), class I chitinase, beta-1,3-glucanase, SAR 8.2, stellacyanin, leucine-rich repeat protein, and auxin-repressed protein were identified. Two CAI genes showed little or no sequence homology to the previously sequenced plant genes. Transcripts of the CAI genes were strongly or preferentially induced in pepper tissues by infection with X. campestris pv. vesicatoria or Phytophthora capsici, and by abiotic elicitor treatment. In particular, most of the CAI genes were strongly induced in pepper tissues by ethephon and methyl jasmonate.

Acetates↗

The exbD2 gene as well as the iron-uptake genes tonB, exbB and exbD1 of Xanthomonas campestris pv. campestris are essential for the induction of a hypersensitive response on pepper (Capsicum annuum).

The tonB, exbB and exbD1 genes of Xanthomonas campestris pv. campestris are essential for ferric iron uptake. In contrast, the exbD2 gene located in the same gene cluster is not essential. Mutational analysis revealed that the ferric-iron-uptake genes tonB, exbB and exbD1 are necessary for the induction of a hypersensitive response (HR) on the nonhost plant pepper (Capsicum annuum) and the induction of typical black rot symptoms on the host plant cauliflower (Brassica oleracea). Again, the exbD2 gene behaved differently. It was found to play a role only in the induction of the HR in pepper but not in the induction of black rot symptoms in cauliflower. Due to the low iron concentration in the plant tissue, the titre of viable bacteria of the ferric-iron-uptake mutants tonB, exbB and exbD1 decreased after leaf infiltration of pepper. The exbD2 mutant, however, which is not impaired in ferric iron uptake, multiplied in the pepper leaf tissue and grew even better than the wild-type strain, probably due to its failure to induce the HR. Nevertheless, the tonB, exbB and exbD1 mutant strains were able to spread systemically in cauliflower.

Bacterial Proteins↗

Production of coumaroylserotonin and feruloylserotonin in transgenic rice expressing pepper hydroxycinnamoyl-coenzyme A:serotonin N-(hydroxycinnamoyl)transferase.

Transgenic rice (Oryza sativa) plants were engineered to express a N-(hydroxycinnamoyl)transferase from pepper (Capsicum annuum), which has been shown to have hydroxycinnamoyl-CoA:tyramine N-(hydroxycinnamoyl)transferase activity, a key enzyme in the synthesis of hydroxycinnamic acid amides, under the control of constitutive maize (Zea mays) ubiquitin promoter. The transgenic rice plants require foliar application of amines to support synthesis of hydroxycinnamic acid amides, suggestive of limiting amine substrates in rice shoots. In addition, when T2 homozygous transgenic rice plants were grown in the presence of amines or phenolic acids, two novel compounds were exclusively identified in the leaves of the transgenic plants. These compounds eluted earlier than p-coumaroyltyramine and feruloyltyramine during HPLC chromatography and were identified as p-coumaroylserotonin and feruloylserotonin by liquid chromatography/mass spectrometry and other methods. To test whether the unpredicted production of serotonin derivatives is associated with the pepper N-(hydroxycinnamoyl)transferase, the substrate specificity of the pepper enzyme was analyzed again. Purified recombinant pepper N-(hydroxycinnamoyl)transferase exhibited a serotonin N-hydroxycinnamoyltransferase (SHT) activity, synthesized p-coumaroylserotonin and feruloylserotonin in vitro, and demonstrated a low K(m) for serotonin. SHT activity was inhibited by 10 to 50 mm tyramine. In addition, SHT activity was predominantly found in the root tissues of wild-type rice in parallel with the synthesis of serotonin derivatives, suggesting that serotonin derivatives are synthesized in the root of rice. This is the first report of SHT activity and the first demonstration, to our knowledge, that serotonin derivatives can be overproduced in vivo in transgenic rice plants that express serotonin N-(hydroxycinnamoyl)transferase.

Acyltransferases↗

Functional analysis of the amine substrate specificity domain of pepper tyramine and serotonin N-hydroxycinnamoyltransferases.

Pepper (Capsicum annuum) serotonin N-hydroxycinnamoyltransferase (SHT) catalyzes the synthesis of N-hydroxycinnamic acid amides of serotonin, including feruloylserotonin and p-coumaroylserotonin. To elucidate the domain or the key amino acid that determines the amine substrate specificity, we isolated a tyramine N-hydroxycinnamoyltransferase (THT) gene from pepper. Purified recombinant THT protein catalyzed the synthesis of N-hydroxycinnamic acid amides of tyramine, including feruloyltyramine and p-coumaroyltyramine, but did not accept serotonin as a substrate. Both the SHT and THT mRNAs were found to be expressed constitutively in all pepper organs. Pepper SHT and THT, which have primary sequences that are 78% identical, were used as models to investigate the structural determinants responsible for their distinct substrate specificities and other enzymatic properties. A series of chimeric genes was constructed by reciprocal exchange of DNA segments between the SHT and THT cDNAs. Functional characterization of the recombinant chimeric proteins revealed that the amino acid residues 129 to 165 of SHT and the corresponding residues 125 to 160 in THT are critical structural determinants for amine substrate specificity. Several amino acids are strongly implicated in the determination of amine substrate specificity, in which glycine-158 is involved in catalysis and amine substrate binding and tyrosine-149 plays a pivotal role in controlling amine substrate specificity between serotonin and tyramine in SHT. Furthermore, the indisputable role of tyrosine is corroborated by the THT-F145Y mutant that uses serotonin as the acyl acceptor. The results from the chimeras and the kinetic measurements will direct the creation of additional novel N-hydroxycinnamoyltransferases from the various N-hydroxycinnamoyltransferases found in nature.

Acyltransferases↗

Anthocyanins, colour and antioxidant properties of eggplant (Solanum melongena L.) and violet pepper (Capsicum annuum L.) peel extracts.

Acetone extracts from eggplant (Solanum melongena L.) and violet pepper (Capsicum annuum L.) peels both belonging to the Solanaceae plant family were characterized with respect to their anthocyanin profiles, colour qualities and antioxidant capacities. According to HPLC-DAD-MS3 analyses the major anthocyanin in eggplant was delphinidin-3-rutinoside, while the predominant pigment in violet pepper was assigned to delphinidin-3-trans-coumaroylrutinoside-5-glucoside. Since virtually all anthocyanins were delphinidin-based, the effect of acylation and glycosylation patterns on colour stability and antioxidant capacity could be assessed. Application of two in vitro-assays for antioxidant capacity assessment revealed that eggplant generally exhibited higher values compared to violet pepper which was ascribed to 3,5-diglycosylated structures predominating in the latter. The higher extent of acylation in violet pepper was reflected by a more purplish colour shade of the extracts, but did not translate into a higher stability against fading which again was attributed to additional glycosyl substitution at C5. These findings support the relevance of structure-related activities of anthocyanins both for understanding food colour and their particular nutritional value.

Anthocyanins↗

[Rapid analysis of aflatoxins in raw peanuts, corn, buckwheat and red pepper by a new mini-column cleanup and HPLC using post-column photochemical derivatization system].

A rapid and clean method for the analysis of aflatoxins (AFs) was developed by using a new column and post-column photochemical derivatization HPLC with fluorescence detection. The new cleanup column consisted of magnesia and basic alumina poured on the top of a commercial multi-functional mini-column. It was extremely effective for the cleanup of AFs from raw peanut, corn, buckwheat and red pepper. Fluorescent substances, which interfered with the analysis of AFs from corn, were completely absorbed at the top of the magnesia layer. Recoveries of AFs (B1, B2, G1, G2) added to raw peanuts, corn, buckwheat and red pepper were over 80% at two levels of fortification (higher level: 10, 3, 10, 3 ng/g, respectively, lower level: 1.0, 0.3, 1.0, 0.3 ng/g, respectively). Coefficients of variation were smaller than 12%, except the lower fortified level for red pepper. Limits of detection for AFs in raw peanuts, corn and buckwheat were 0.3 ng/g for B1 and G1, and 0.1 ng/g for B2 and G2. Those in red pepper were 0.5 ng/g for B1, B2, G1 and G2.

Aflatoxins↗

An alternative method to screen for pepper spray residue.

A method was developed to screen for pepper spray residue using instruments and methods other than those techniques commonly employed to analyze chemical residue (i.e.. gas chromatography mass spectrometry-GCMS or liquid chromatography mass spectrometry-LCMS). The method employed gas chromatography (GC), thin layer chromatography (TLC), and diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) to screen for dried pepper spray stains. Pepper sprays from nine different manufacturers were investigated. Capsaicin and dihydrocapsaicin were identified and unique IR reflectance spectra are presented. An additional five compounds were presumptively found. Results showed that a particular stain could be characterized as a pepper-based stain.

Journal Article↗

[Effects of different application rates of humic acid compound fertilizer on pepper and its mechanism of anti-senility and incremental yield].

The effects of different application rates of humic acid compound fertilizer (HA) on the yield and physiological characteristics of pepper (Capsicum frutescens L.) were studied in this paper. The results showed that different application rates of (HA) could affect the physiological activities and yield of pepper. The nitrate reductase activity (NRA) and peroxidase (POD) activity were improved with increasing application rate, but the change rate of NRA was reduced gradually. POD activity in inorganic compound fertilizer treatment was improved significantly when the application rate of fertilizer was higher than 0.3 g.kg-1, but that in HA treatment changed smoothly. Superoxide dismutase (SOD) activity was improved significantly when the application rate of fertilizer was lower than 0.2 g.kg-1, and decreased gradually when the application rate was higher than 0.2 g.kg-1. The change of transpiration rate was consistent with that of SOD activity. The relationship between the application rate of fertilizer and the yield of pepper could be described with quadratic curve regression model. According to the equation, the highest application rate of HA should be 0.27 g.kg-1, and the highest yield of pepper could reach 165.22 g per pot.

Capsicum↗

A method of high frequency virus-induced gene silencing in chili pepper (Capsicum annuum L. cv. Bukang).

Using a tobacco rattle virus (TRV)-based virus-induced gene silencing (VIGS) system, expression of phytogene desaturase (PDS) and ribulose-1,5-bisphosphate carboxylase small-subuit (rbcS) genes was suppressed in Nicotiana benthamiana and pepper plants (Capsicum annuum L. cv. Bukang). The silenced phenotypes of pale yellow (rbcS), and photobleached leaves (PDS), were invariably obvious 2 weeks after inoculation with the TRV-based vector. In a parallel experiment, the same set of genes was silenced in N. benthamiana and yielded identical phenotypes to pepper 1 week after inoculation. Northern blot analyses showed that the endogenous levels of CarbcS and CaPDS transcripts were dramatically reduced in the silenced leaf tissues. These observations confirm that the silenced phenotype is closely correlated with the pattern of tissue expression. To our knowledge, this is the first high frequency VIGS method in pepper plants. It should provide a tool for large-scale gene silencing studies in pepper functional genomics.

Agrobacterium tumefaciens↗

Biological control of Aphis gossypii on pepper plant using Coccinella algerica.

To calibrate biological control in pepper plant against Aphis gossypii using Coccinella algerica different experimentation were made. The consumption of C. algerica during the larval stage was enumerated in laboratory conditions. The densities (1, 4, 8 larvae/plant) to L2 stage of C. algerica necessarily to decrease aphid's populations were determined in plant pepper and the daily consumption in the pepper plant at the different densities was calculated. The density with 8 larvae at semi natural conditions is higher significantly. In protected crop of pepper, different releases to C. algerica were made with eggs, second stage larva and with ladybird before infestation with A. gossypii. A strategy to introduce L2 stage is preferred than the release with punters of C. algerica at different densities. At the same, the Introduce L2 stage is compared to release the C. algerica ladybird before the infestation of A. gossypii in the protected crops. The activity of ladybird has been maintained in the protected crop and it was shown when the infestation appeared.

Anacardiaceae↗

Identification of irradiated pepper with the level of hydrogen gas as a probe.

A novel method to detect whether or not a particular pepper has been irradiated has been developed which is based on the fact that H2 is formed in organic substances irradiated with ionizing radiation. Following gamma irradiation, black and white peppers were ground to powder in a gastight ceramic mill. By gas-chromatographic analysis of the gas in the mill, we observed that H2 had been released from the irradiated pepper grains. Curves plotting the H2 content vs storage time at storage temperatures of 7, 22, and 30 degrees C showed that the higher the temperatures, the smaller the H2 content, and that identification of irradiated pepper was possible for 2-4 months after 10 kGy irradiation.

Chromatography, Gas↗

[The value of Polish field grown sweet pepper cultivars for freezing and pickling].

Raw and blanched sweet pepper and frozen goods and pickles produced of blanched and non-blanched vegetable were evaluated after 6-month storage, the following cultivars of Polish origin being used in the screening: Bryza, Ino, Jantar, Kano, Kujawianka, Passat, Sono, and Zefir. The compared cultivars differed by the level of analysed indices. The variability range of dry matter content was 25%, of sugars 23%, of starch 97%, of protein 25%, of mineral constituents 24%, and of vitamin C 40%. The blanching of the raw material reduced the content of sugar by 11%, raw protein by 8%, mineral constituents by 14%, and vitamin C by 18%. The freezing and 6-month storage of frozen sweet pepper did not significantly affect its chemical composition, except vitamin C whose level was decreased by 17% on the average in the case of freezing non-blanched raw material, and by 22% when blanched sweet pepper was used. The pickling and storage of pickles significantly reduced the content of vitamin C, the losses amounting to 58% in products of non-blanched and to 67% in these of blanched fruit. The lossesin protein content were relatively small while and increase in the content of dry matter, sugars, and ashy constituents was due to additives in the marinade. The process of blanching sweet pepper before freezing ensure the preservation of proper smell and taste. Hence, this treatment has to be regarded as indispensable prior to the longterm storage of frozen vegetables. In the case of pickling the blanching has no favourable effect on the organoleptic qualities. The highest organoleptic values was in pickles and frozen goods of Kujawianka, the scores being 4.5 each, however no statistical differences were found for products of Kano, Sono, Bryza, and Zefir.

Ascorbic Acid↗

Microbial and mycotoxic contamination of peppers and food safety.

Twelve random samples of pepper powders were analyzed for contamination by microorganisms of public health significance (bacteria, moulds) and for aflatoxin B1 concentration. Black peppers were more heavily contaminated than white peppers but the most important bacteria causing foodborne diseases were not isolated; The mould flora was dominated by xerophilic species, particularly Aspergillus fumigatus, A. flavus, A. niger and A. ochraceus. These potentially pathogenic species can be dangerous for predisposed patients. The most striking result concerns aflatoxin B1 which was detected in every sample analyzed and could not always be correlated with the occurrence of a mycotoxinogenic strain of Aspergillus, suggesting the use of a sterilizing treatment previously. So, the detection of aflatoxin B1 must be a necessary criterion to be added to microbial criteria to appreciate the quality of peppers.

Aflatoxin B1↗

[Allergy associated with pepper and latex: new cross reaction?].

The aim of this study was to point out the frequency of associated sensitization to rubber latex and to sweet pepper. The study included three populations of patients: Patients allergic to rubber latex, patients sensitized to rubber latex and patients without any clinical symptoms for rubber latex and sweet pepper (negative controls). We pointed out a high frequency of patients sensitized to sweet pepper among the patients allergic or sensitized to rubber latex which lead to the hypothesis of a cross reaction between rubber latex and sweet pepper.

Adolescent↗

Resistance to tobamoviruses in transgenic tobacco plants expressing the coat protein gene of pepper mild mottle virus (Korean isolate).

Red pepper, one of the most important vegetable crops in Korea, is severely affected by viral diseases causing 20-50% reduction in product yield. A pepper strain of tobacco mosaic virus (TMV-p) is the most common virus in red pepper. To study the molecular structure of the TMV-p virus, we generated cDNA clones of the viral genome. Partial sequencing of a few cDNA clones revealed that TMV-p shares a 98% identity at the nucleotide level with the Spanish isolate of pepper mild mottle virus (PMMV-s). This suggests that TMV-p should be reclassified as the Korean isolate of PMMV (PMMV-k). The coat protein (CP) gene together with the 3' untranslated region of the PMMV-k virus was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) using oligomers deduced from the sequence of PMMV-s. The sequence analysis of the CP gene and the 3' untranslated region further confirmed that PMMV-k is highly related to PMMV-s. The CP gene and the 3' untranslated region of PMMV-k were cloned into a plant expression vector and the construct was introduced into tobacco plants. The transgenic plants expressing the PMMV-k CP gene were delayed in developing systemic disease or failed to develop symptoms at all after inoculation with PMMV-k. Delay of symptoms was also observed when the plants were inoculated with TMV-OM which shares a 74% homology with PMMV-k in the amino acid sequence of the CP region. In a local lesion host, the CP expressing plants exhibited a greatly reduced number of necrotic lesions as compared to control plants after inoculation with TMV-OM. Our results show that CP-mediated viral resistance is readily applicable in the case of PMMV-k and can provide resistance to other viruses in the tobamovirus group.

Amino Acid Sequence↗

CaMYB121-CaABF2 negative feedback loop modulates CaNHX2 expression to confer salt tolerance in pepper.

Salt stress is a major abiotic factor that severely restricts pepper (Capsicum annuum) production. Although abscisic acid (ABA) is vital for salt tolerance, the transcriptional regulatory networks governing ABA-mediated salt defense remain largely unknown. Here, we uncovered a negative feedback loop between CaMYB121 and CaABF2.1/2 that modulates the expression of CaNHX2.1/2/3, thereby enhancing salt tolerance in pepper plants. RNA-seq analysis revealed that CaMYB121 displayed an expression pattern consistent with that of CaNHX2 after salt treatment. Silencing CaMYB121 markedly reduced salt tolerance and inhibited root growth. Mechanistically, CaMYB121 directly binds to the CaNHX2 promoter to activate transcription, thereby promoting salt resilience. Salt stress also robustly triggered ABA signaling genes, with CaABF2.1/2 displaying expression patterns closely mirroring those of CaMYB121. Transient silencing of CaABF2.1/2 results in phenotypes similar to those observed with CaMYB121 suppression. Notably, CaMYB121 activates CaABF2.1/2 transcription by binding to its promoters, whereas CaABF2.1/2 represses CaMYB121 expression by directly targeting its promoter, forming a self-regulating feedback loop that prevents excessive defense activation. Collectively, our findings reveal a CaMYB121-CaABF2 feedback circuit that dynamically balances growth and defense to optimize salt tolerance in pepper plants.

Salt Tolerance↗

Genome-Wide Identification, Phylogenetic Analysis, and Expression Pattern of Polyamine Biosynthesis Gene Family in Pepper.

Polyamines (PAs), including putrescine, spermidine, spermine, and thermospermine, play essential roles in plant growth, development, and responses to stress. However, the structure and function of PA biosynthetic genes in pepper remain poorly characterized. This study aimed to identify PA biosynthesis genes in the pepper genome using bioinformatics approaches and to assess their expression under various stress conditions. A total of 16 PA biosynthesis-related genes were identified, representing members of the arginine decarboxylase (ADC), ornithine decarboxylase (ODC), agmatine iminohydrolase (AIH), N-carbamoylputrescine amidohydrolase (CPA), S-adenosylmethionine decarboxylase (SAMDC), spermidine synthase (SPDS), spermine synthase (SPMS), and ACAULIS5 (ACL5) gene families. These genes encode proteins with an average molecular weight of approximately 40 kDa, primarily localized in the mitochondria and cytoplasm. Promoter analysis revealed multiple cis-acting elements associated with stress and phytohormone responsiveness. Gene expression was induced by various abiotic stresses, including saline-alkaline, drought, heat, cold, and hydrogen peroxide, as well as by phytohormones such as abscisic acid, ethylene, salicylic acid, auxin, and gibberellin. Overall, this study provides a comprehensive analysis of PA biosynthesis genes in pepper and highlights their potential roles in stress adaptation and hormone signalling, offering a foundation for further exploration of PA-mediated stress tolerance mechanisms.

Capsicum↗