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Development of a CAPS marker for the Pvr4 locus: a tool for pyramiding potyvirus resistance genes in pepper.

The Pvr4 resistance gene in pepper confers a complete resistance to the three pathotypes of potato virus Y (PVY) and to pepper mottle virus (PepMoV). In order to use this gene in a marker-assisted selection (MAS) program and to permit the pyramiding of several potyvirus resistance genes in the same cultivar, tightly linked amplified fragment length polymorphism (AFLP) markers were obtained by the bulked segregant analysis method. Eight linked AFLP markers were mapped in an interval from 2.1 +/- 0.8 to 13.8 +/- 2.9 cM around this locus. The closest codominant AFLP marker was converted into a codominant CAPS (cleaved amplified polymorphic sequence) marker using data from the alignment of the two allele sequences. We have further characterized the relevance of the CAPS marker for MAS programs in different pepper breeding lines.

Capsicum↗

Use of SSR markers to complement tests of distinctiveness, uniformity, and stability (DUS) of pepper (Capsicum annuum L.) varieties.

This study was carried out to assess the potential of SSR markers for variety identification by comparing SSR markers and morphological traits in tests of distinctiveness, uniformity, and stability (DUS) of pepper (Capsicum annuum L.) varieties. Twenty-seven SSR markers were polymorphic in 66 pepper varieties, revealing a total of 89 alleles. Average polymorphism information content (PIC) value was 0.529, ranging from 0.03 to 0.877. Cluster analysis of the band patterns separated the varieties into three groups corresponding to varietal types. Morphological trait-based clustering showed some degree of similarity to dendrogram topologies based on the SSR index. However, no significance correlation was found between the SSR and morphological data. SSR markers could be used to complement a DUS test of a candidate variety and to select complimentary varieties by pre-screening existing varieties in the context of protecting new varieties of pepper.

Capsicum↗

Cumulative trauma disorders in betel pepper leaf-cullers visiting a rehabilitation clinic: experience in Taitung.

BACKGROUND: Betel pepper (Piper betle L.) cultivation is an important agricultural industry in Taitung, Taiwan, and culling leaves is very labor-intensive. This case study compares the proportion of cumulative trauma disorders (CTDs) between cullers and those with other occupations. METHODS: Patients with musculoskeletal disorders in the rehabilitation clinic of a local hospital in Taitung were enrolled. This all female cohort was divided into a culler group (betel pepper cullers, n = 20), and a non-culler group (other occupations, n = 47). Three cullers were interviewed, and were also recorded to elucidate the related ergonomics. Patients were diagnosed using plain radiography and ultrasonography. RESULTS: The act of culling involves an overhead internal rotation of both shoulders with extended elbows while standing, followed by wrist flexion and forearm pronation. Flexing of the fingers is also required by the tools, 'iron nails' fitted onto both thumbs. The proportions of patients with shoulder impingement syndrome (SIS) and carpal tunnel syndrome (CTS) were significantly higher among cullers than non-cullers (0.45 vs. 0.15, p = 0.011 and 0.40 vs. 0.06, p = 0.002, respectively). Furthermore, the total frequency of CTDs displayed a positive linear correlation with employment duration (r = 0.618, p = 0.004). CONCLUSIONS: Proportions of occupational SIS and CTS were higher among betel pepper cullers than those with other occupations. These CTDs may have resulted from a prolonged static posture and repetitive motions during culling.

Adult↗

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↗

Development of SCAR markers for early identification of cytoplasmic male sterility genotype in chili pepper (Capsicum annuum L.).

We previously used Southern blot analysis to detect restriction-length polymorphisms between male fertile and cytoplasmic male sterile (CMS) cytoplasms at the coxII and atp6 loci of the mtDNA of Capsicum annuum L. Two copies of atp6 were found in each male fertile and CMS pepper lines. Interestingly, one of the copies of atp6 in CMS pepper was a 3'-truncated pseudogene. The open reading frame of the coxII gene was the same in the fertile (N-) and CMS (S-) lines. However, the nucleotide sequence in the S-cytoplasm diverged from that in the N-cytoplasm 41 bp downstream of the stop codon. To develop CMS-specific sequence-characterized amplified region (SCAR) markers, inverse PCR was performed to characterize the nucleotide sequences of the 5' and 3' flanking regions of mitochondrial atp6 and coxII from the cytoplasms of male fertile (N-) and CMS (S-) pepper plants. Based on these data, two CMS-specific SCAR markers, 607 and 708 bp long, were developed to distinguish N-cytoplasm from S-cytoplasm by PCR. The CMS-specific PCR bands were verified for 20 cultivars containing either N- or S-cytoplasm. PCR amplification of CMS-specific mitochondrial nucleotide sequences will allow quick and reliable identification of the cytoplasmic types of individual plants at the seedling stage, and assessment of the purity of F1 seed lots. The strategy used in this report for identifying CMS-specific markers could be adopted for many other crops where CMS is used for F1 seed production.

3' Untranslated Regions↗

[Novel application of FTIR to the analysis of sweet pepper seeds and tomato seeds with space mutation].

Comparative study of the sweet pepper and tomato seeds lines with space mutation and corresponding conventional seeds was carried out with FTIR, in an attempt to search for the influence of space environment upon mutation. The similarities among space seeds lines are: the absorbance at near 1,160-1,061 cm(-1) increased. This result shows that the stretching vibration of C-O on the structure of carbohydrates was enhanced by space mutation. The special mutations are: (1) The absorbance at 2,854 cm(-1) of space sweet pepper was enhanced, showing that the stretching vibration of CH2 was stronger; (2) The absorbance at 1,160 cm(-1) of cherry tomato seeds lines was increased notably, indicating that the stretching vibration of C-O on the structure of carbohydrates was stronger. The results show that the space environment could notably increase the carbohydrates in the tomato seeds. The space sweet pepper and tomato seeds lines, which produced similar mutation, could be influenced by microgravity in space. The special mutations are the results of the influence of universe radiation in space.

Capsicum↗

Polymorphism of pepper (Capsicum annuum L.) seed proteins studied by two-dimensional electrophoresis with immobilized pH gradients: methodical and genetic aspects.

A population of ten morphologically similar inbred lines of pepper (Capsicum annuum L.) has been investigated for polymorphism of seed proteins by two-dimensional (2-D) electrophoresis with immobilized pH gradients. To reveal as much variation as possible, both the water- and the urea/detergent-soluble protein fraction were electrophoretically analyzed and subsequently visualized by silver staining. The 2-D patterns were subjected to computer analysis to be able to establish genetic variation. A high number of the seed proteins were found to be variable as to presence/absence: these were 68 out of 184 reproducible water-soluble proteins and 34 out of 419 reproducible urea/detergent-soluble proteins. Comparison of the 2-D data of the water-soluble and the urea/detergent-soluble proteins, which represent the biggest part of all extractable seed proteins, showed that both protein fractions have proteins in common, but the variable proteins found in both fractions were non-identical. The difference of variability scored in both solubility fractions was discussed. Genetic distances between all pairs of inbred pepper lines were calculated and a genetic tree was constructed. A correlation analysis was carried out to correct for genetic linkage and for secondary modifications, to have a more proper estimate of genetic distances. In both cases the dendrograms showed two distinct genetic groups of five inbred lines. This electrophoretic study was done in order to utilize the genetic distance data in breeding for heterosis. The genetic distance data presented will be used to validate the assumption that there is a higher chance to achieve better hybrid performance when the genetic distance between the parents is as great as possible.(ABSTRACT TRUNCATED AT 250 WORDS)

Breeding↗

Association of protein polymorphism among pepper (Capsicum annuum L.) inbred lines with agronomic performance of their crosses.

The genetic variation of 10 morphologically similar pepper (Capsicum annuum L.) inbred lines has been analyzed by two-dimensional electrophoresis with immobilized pH gradient (IPG-DALT) of seed proteins. For all pairs of inbred lines genetic distance indices were calculated on the basis of the IPG-DALT analysis and a genetic tree was constructed. The protein polymorphism data of the 10 inbred lines were integrated into a pepper breeding program to validate the assumption that there is a higher chance of achieving better hybrid performance when the genetic distance between the parents is as great as possible. Field trials were performed in Turkey and consisted of a total of 27 crossings based on 9 inbred lines. Fifteen hybrids exceeded the crop yield of the better parent and 8 additional hybrids exceeded the mean crop yield of both parents. The genetic distance indices of the parental inbred lines based on protein polymorphism data were not significantly correlated to heterosis performance of the experimental hybrids (r = 0.20), indicating that protein polymorphism data add little to the prediction of single cross hybrid performance. Nevertheless, protein polymorphism data support the breeder in being more effective in finding out the best inbred combinations, because most of the less promising crossings can be omitted from the expensive and time-consuming performance tests. Those hybrids whose distance indices exceeded the mean of all 27 distance values (0.52) were superior to the mean value of all heterosis performances by 19%.(ABSTRACT TRUNCATED AT 250 WORDS)

Capsicum↗

Evaluation of mutagenic activity in an extract of pepper tree stem bark (Schinus terebinthifolius Raddi).

An extract (decoction) from pepper tree stem bark (Schinus terebinthifolius Raddi) is widely used in Brazil as a topical antiinflammatory agent and to cicatrize wounds. The extract contains catechin, tannins, terpenes, flavonoids, and saponins; of these components, both mutagenic potential and antioxidant properties have been ascribed to flavonoids. The mutagenicity of some flavonoids is believed to be associated with the formation of reactive oxygen species and seems to depend on the number and position of hydroxyl groups. In the present study, we evaluated an extract of S. terebinthifolius in a series of cell-free and bacterial assays in order to determine its genotoxic potential. The extract was negative in a cell-free plasmid DNA test, indicating that it did not directly break DNA. Positive results, however, were obtained in the SOS chromotest, in a forward mutagenesis assay employing CC104 and CC104mutMmutY strains of Escherichia coli, and in the Salmonella reversion assay, using strains TA97, TA98, TA100, and TA102. All the bacterial tests were performed without exogenous metabolic activation due to the topical use of this preparation. The results indicate that pepper tree stem bark extract produces DNA damage and mutation in bacteria, and that oxidative damage may be responsible for the genotoxicity.

Anacardiaceae↗

Comprehensive two-dimensional gas chromatography/mass spectrometric analysis of pepper volatiles.

The headspace compositions of 13 pepper and peppercorn samples of different species, colloquially also referred to as pepper, were analyzed, and more than 300 compounds were tentatively characterized by means of comprehensive two-dimensional gas chromatography in tandem with flame ionization detection, quadrupole mass spectrometric detection and time-of-flight mass spectrometric detection (GC x GC-FID, GC x GC/qMS and GC x GC/TOFMS, respectively). The analysis of volatile organic compounds (VOCs) was performed after solid-phase microextraction (SPME) using a 75-microm PDMS/DVB fibre. Fingerprint comparison between the three techniques permitted peaks to be assigned in the GC x GC-FID experiment based on the analogous MS analysis, taking into account retention shifts arising from method variations. When using GC x GC/TOFMS, about five times more peaks were identified than in GC x GC/qMS. Retention indices for all peaks were calculated in the bi-dimensional column set comprising of a 5% phenyl polysilphenylene-siloxane primary column and a polyethylene glycol second column. The spectra obtained by both mass detection techniques (qMS and TOFMS) give very similar results when spectral library searching was performed. The majority of the identified compounds eluted as pure components as a result of high-resolution GC x GC separations, which significantly reduces co-elution, and therefore increases the likelihood that pure spectra can be obtained. The differences between TOFMS and qMS (in fast scanning mode) spectra were generally small. Whilst spectral quality and relative ion ratios across a narrow peak (e.g. w(b) approximately 100-150 ms) do vary more for the fast peaks obtained in GC x GC/qMS operation, than with TOFMS, in general adequate spectral matching with the library can be achieved.

Gas Chromatography-Mass Spectrometry↗

A comparison of pepper mottle virus with potato virus Y and evidence for their distinction.

Pepper mottle virus (PepMOV) was identified as a distinct potyvirus infecting peppers in Arizona and Florida in the 1970's. The distinction of PepMoV from potato virus Y (PVY) has recently been challenged on the basis of sequence comparisons of the coat proteins and of the 3' nontranslated regions of the viral RNAs. We summarize the biological, cytological, serological, and in vitro translational studies which compare the apparent differences, and also similarities, between PepMoV and PVY. We conclude that although PepMoV may be more closely related to PVY than to other known potyvirus, PepMoV should be maintained as a separate virus on the basis of its distinctive characteristics.

Plant Viruses↗

Is pepper mottle virus a strain of potato virus Y?

On the basis of serological properties and host plant reactions pepper mottle virus (PepMoV) has been classified as a potyvirus related to, but distinct from, other pepper-infecting potyviruses, potato virus Y (PVY) and tobacco etch virus (TEV). Recent amino acid and nucleotide sequence data show that PepMoV is more closely related to PVY than previously assumed. PepMoV shows a high degree of homology to various PVY strains in both the coat protein and the 3' non-translated sequences, while unrelated potyviruses are generally less homologous in these regions. Detailed coat-protein amino acid sequence and 3' non-translated region (3' NTR) nucleotide sequence comparisons described in this paper confirm the close relationship between PepMoV and PVY and it is concluded that the isolate sequenced indeed represents a strain of PVY. Sequence data for several strains of PVY gave two groups with closer relationships among strains in a group than between groups.

Amino Acid Sequence↗

Molecular cloning and expression of the hot pepper ERabp1 gene encoding auxin-binding protein.

The 22 kDa auxin-binding proteins in higher plants have received considerable attention as candidates for an auxin receptor. A cDNA clone Ca-ERabp1 of hot pepper (Capsicum annum) was isolated using the oligonucleotides as PCR primers. The cDNA codes for a polypeptide related to the major 22 kDa auxin-binding protein from maize and Arabidopsis ERabp1. The deduced amino acid sequence contains an endoplasmic reticulum retention signal, the KDEL sequence located at the C-terminal end, and has two possible auxin-binding sites, HRHSCE and YDDWSVPHTA conserved sequences. Northern hybridization analysis revealed that the Ca-ERabp1 gene is differentially expressed in total RNA isolated from different organs of a pepper plant, showing the highest level of expression in fruits but barely detectable in leaves and roots.

Amino Acid Sequence↗

The psbL gene from bell pepper (Capsicum annuum): plastid RNA editing also occurs in non-photosynthetic chromoplasts.

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.

Amino Acid Sequence↗

Characterization of a family of genes encoding a fruit-specific wound-stimulated protein of bell pepper (Capsicum annuum): identification of a new family of transposable elements.

Using a fruit-specific cDNA as a probe we isolated and sequenced the two corresponding homologous genes (Sn-1 and Sn-2) of the bell pepper (Capsicum annuum) genome. Both genes have a single intron and numerous unusual long inverted repeat sequences. The introns share 87% homology and Sn-2 contains one 450 bp additional sequence with structural features of a transposable element, which is highly repetitive in the bell pepper genome. Surprisingly, analysis in data banks showed that genes encoding the potato starch phosphorylase (EC 2.4.1.1) and patatin contain a similar element, named Alien, in their 5'-upstream region. Alien elements are characterized by a conserved 28 bp terminal inverted repeat (TIR), small size, high AT content, potential to form stable DNA secondary structures and they have probably been inserted in TA target sites. Interestingly, the TIR of the Alien elements shares high homology with sequences existing in the TIR of extrachromosomal linear pSKL DNA plasmid of Saccharomyces kluyveri. Northern blot analyses detected Sn-1 transcripts principally in the red fruit whereas no Sn-2 transcripts were detected in neither of the samples monitored. Western blot analyses detected a 16.8 kDa Sn protein principally in the ripe red fruit and wounded areas of green unripe fruit. A comparison of the deduced amino acid sequence of Sn-1 with protein sequences in data banks revealed a significant homology with proteins likely involved in the plant's disease resistance response. Analyses at the subcellular level showed that Sn-1 is localized in the membrane of vacuoles.

Amino Acid Sequence↗

Differential ethylene-inducible expression of cellulase in pepper plants.

Ethylene promotes the abscission of leaves and the ripening of fruits in pepper plants, and in both events an increase in cellulase activity is observed. However, two enzyme isoforms (pI 7.2 and 8.5, respectively) are differentially involved in the two physiological phenomena. The pI 8.5 form has been purified from ripe fruits. It is a glycoprotein with an apparent molecular mass of 54 kDa. Two short peptides were sequenced and a very high homology to a tomato cellulase was observed. Polyclonal antibodies, raised against the purified enzyme, have allowed us to demonstrate that the observed ethylene-induced increase in cellulase activity is paralleled by de novo synthesis of protein. Three cDNAs (CX1, CX2 and CX3), encoding different cellulases, were obtained and characterized and their expression investigated. Accumulation of all three mRNAs is induced by ethylene treatment, though to different levels. CX1 is mainly expressed in ripe fruits while CX2 is especially found in abscission zones. CX3 accumulates at very low levels in activated abscission zones. Comparisons with other known cellulases demonstrate clear heterogeneity within the higher plant cellulases. Differences in ethylene inducibility and molecular structure suggest different physiological roles for cellulase in pepper plants.

Amino Acid Sequence↗

Identification and structural characterization of further DNA elements in the potato and pepper genomes homologous to the transposable element-like insertion Tst1.

The molecular cloning and nucleotide sequence of elements from potato and pepper that are related to the recently identified Tst1 element are described. Sequence analysis reveals considerable conservation of sequences internal to both the Tst1 element and two of the related elements identified here. In six potato clones analysed, the 11 bp inverted repeat first identified in the Tst1 element is conserved. Several of the elements are flanked by an 8 bp direct repeat. DNA fragments which were amplified from several pepper genomes by polymerase chain reaction (PCR) amplification using the inverted repeat as sequence primers also display considerable conservation of sequences internal to the Tst1 element. These data further support the possibility that Tst1 is a non-autonomous transposable element and that Tst1 might be the first example of a transposable element which occurs in several genera of solanaceous plants.

Base Sequence↗

Alteration of apparent viscosity of irradiated pepper--a tool for semi-quantitative estimation of irradiation dose.

The feasibility of using apparent viscosity (eta a) as a method for detecting the occurrence of previous irradiation of pepper was studied. Apparent viscosity of heat-treated suspensions of white and black pepper, nonirradiated or irradiated with different doses of ionising radiation (gamma), was measured under different "shear rates". Results of previous research were therefore expanded and their usefulness examined; low shear rate conditions were found to be preferable for the detection and semi-quantitative evaluation of irradiation doses. The experimental methodology for semi-quantitative estimation was developed and its scope and limitations are presented.

Dose-Response Relationship, Radiation↗