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Towards the saturation of the pepper linkage map by alignment of three intraspecific maps including known-function genes.

Three populations composed of a total of 215 doubled haploid lines and 151 F2 individuals were used to design an intraspecific consensus map of pepper (Capsicum annuum L.). The individual maps varied from 685 to 1668 cM with 16 to 20 linkage groups (LGs). The alignment of the three individual maps permitted the arrangement of 12 consensus major linkage groups corresponding to the basic chromosome number of pepper and displaying a complex correspondence with the tomato map. The consensus map contained 100 known-function gene markers and 5 loci of agronomic interest (the disease-resistance loci L, pvr2, and Pvr4; the C locus, which determines capsaicin content; and the up locus, controlling the erect habit of the fruits). The locations of three other disease-resistance loci (Tsw, Me3, and Bs3) and the y locus, which determines the yellow fruit colour, were also found on this consensus map thanks to linked markers. Here we report on the first functional detailed map in pepper. The use of candidate gene sequences as genetic markers allowed us to localize four clusters of disease-resistance gene analogues and to establish syntenic relationships with other species.

Capsicum↗

A comparison of sole carbon source utilization patterns and phospholipid fatty acid profiles to detect changes in the root microflora of hydroponically grown crops.

Sole carbon source utilization (SCSU) patterns and phospholipid fatty acid (PLFA) profiles were compared with respect to their potential to characterize root-inhabiting microbial communities of hydroponically grown crops. Sweet pepper (Capsicum annum cv. Evident), lettuce (Lactuca sativa cv. Grand Rapids), and four different cultivars of tomato (Lycopersicon esculentum cvs. Gitana, Armada, Aromata, and Elin) were grown in 1-L black plastic beakers placed in a cultivation chamber with artificial light. In addition to the harvest of the plants after 6 weeks, plants of one tomato cultivar, cv. Gitana, were also harvested after 4 and 8 weeks. The cultivation in this study was performed twice. Principal component analysis was used to analyze the data. Both characterization methods had the ability to discriminate between the root microflora of different plant species, cultivars, and one tomato cultivar at different ages. Differences in both SCSU patterns and PLFA profiles were larger between plant species than between cultivars, but for both methods the largest differences were between the two cultivations. Still, the differences between treatments were always due to differences in the same PLFAs in both cultivations. This was not the case for the SCSU patterns when different plant ages were studied. Furthermore, PLFA profiles showed less variation between replicates than did SCSU patterns. This larger variation observed among the SCSU data indicates that PLFA may be more useful to detect changes in the root microflora of hydroponically grown crops than the SCSU technique.

Capsicum↗

Ultrastructural rRNA localization in plant cell nucleoli. RNA/RNA in situ hybridization, autoradiography and cytochemistry.

The distribution of ribosomal transcripts in the plant nucleolus has been studied by non-isotopic in situ hybridization in ultrathin Lowicryl K4M sections and by high-resolution autoradiography after labelling with tritiated uridine. In parallel, cytochemical techniques were applied to localize RNA on different plant nucleolar components of Allium cepa L. root meristematic cells and Capsicum annuum L. pollen grains. For RNA/RNA in situ hybridization, several biotinylated single-stranded ribosomal RNA probes were used for mapping different fragments of the 18 S and the 25 S rRNA gene transcribed regions. Ribosomal RNAs (from pre-rRNAs to mature 18 and 25 S RNAs) were found in the nucleolus, in the dense fibrillar (DFC) and granular components (GC). Hybridization signal was found at the periphery of some fibrillar centres (FCs) with probes recognizing both 18 and 25 S rRNA sequences. A quantitative study was performed to analyze the significance of this labelling. Incorporation of tritiated uridine into roots was carried out and, later, after a long time-exposure, autoradiography revealed the presence of newly synthesized RNA mainly in the DFC and at the periphery of the FCs. The presence of RNA in these areas was also confirmed by the cytochemical techniques used in this study. Taken together, these data favour the hypothesis that transcription can begin at the periphery of the FCs, although we cannot exclude the possibility that the DFC plays a role in this process.

Allium↗

Changes of lipoxygenase and fatty acid hydroperoxide lyase activities in bell pepper fruits during maturation.

Developmental changes in fatty acid hydroperoxide lyase (HPO lyase) and lipoxygenase (LOX) during the maturation of bell pepper fruits (Capsicum annuum L. cv. Kyonami) were examined by means of activity measurements, immunological detection of both the enzymes, and analysis of the volatile compounds formed upon homogenization of the fruits. Both the enzyme activities decreased with maturation, and immunological studies showed that the amounts of the enzymes concomitantly decreased. The amounts of six-carbon aldehydes and alcohols formed from bell pepper fruits upon homogenization also decreased during maturation, and with the fully ripened red fruits, these volatile compounds were hardly detectable. These results suggest that the major factor contributing to the changes in the composition of volatile compounds during the maturation of bell pepper fruits was changes in the amounts of HPO lyase and LOX.

Alcohols↗

Household treatment for "chile burns" of the hands.

In New Mexico, chile peppers (Capsicum annum) are prepared by roasting and manually removing the skin from the fruit. Peeling is often done barehanded and may cause prolonged burning pain, irritation, and erythema but not vesication. In a survey of elderly Hispanic women, treatment with oils or cool tap water were frequently used home remedies. Twenty female subjects immersed their hands in a standardized slurry of green chile for 40 minutes, afterwards one hand was placed in cool tap water and the other in vegetable oil for a total of 75 minutes. Pain was scored using a visual analog scale while the hands were immersed in the chile slurry, test baths, and after drying. The difference in pain score was calculated for each subject. Analysis was by pooled regression. Cool tap water immersion initially provided more relief while vegetable oil provided better long-term relief from the pain of "chile burns".

Adult↗

Amelioratory effect of dietary ingestion with red bell pepper on learning impairment in senescence-accelerated mice (SAMP8).

The effect of dietary red bell pepper (Capsicum annuum L.) on learning performance was studied in the senescence-accelerated mouse (SAM). An experimental diet, which contained 20% (w/w) lyophilized powder of red bell pepper, was fed to SAMP8 mice. The mice that received the experimental diet showed much better acquisition in passive avoidance tasks as compared with a control group given a common diet. This indicated that the dietary ingestion of red bell pepper ameliorated the learning impairment in SAMP8.

Aging↗

Four-week supplementation with a natural dietary compound produces favorable changes in body composition.

The purpose of this study was to determine whether a natural dietary supplement produced favorable changes in body composition during a 4-week diet- and-exercise program. The active compound contains a patented combination of chromium picolinate, inulin, capsicum, L-phenylalanine, and other lipotropic nutrients. A double-blind, weight-loss intervention design was used. Participants were randomly assigned to either a diet/exercise/supplement group (n = 56) or a diet/exercise/placebo group (n = 67). Caloric intake was reduced to 1500 kcal/d and participants walked for 45 minutes, 5 days a week, to attain between 60% and 80% of predicted maximal heart rate. Analysis of covariance (ANCOVA) showed significant differences (P < .05) between groups in percent body fat, fat mass, and fat-free mass; no significant differences were found (P > .05) in body weight, body mass index, or energy intake. Independent t tests showed no significant differences (P > .05) in diet composition between groups. Results indicate that the addition of a natural dietary supplement during a 4-week diet-and-exercise weight-loss program accelerates the rate of body fat loss and helps maintain fat-free mass (lean tissue), thereby producing favorable changes in body composition.

Adipose Tissue↗

A CAPS marker to assist selection of tomato spotted wilt virus (TSWV) resistance in pepper.

The hypersensitive resistance to tomato spotted wilt virus (TSWV) in pepper is determined by a single dominant gene (resistant allele: Tsw) in several Capsicum chinense genotypes. In order to facilitate the selection for this resistance, four RAPD (among 250 10-mer primers tested) were found linked to the Tsw locus using the bulked segregant analysis and 153 F2 individuals. A close RAPD marker was converted into a codominant cleaved amplified polymorphic sequence (CAPS) using specific PCR primers and restriction enzymes. This CAPS marker is tightly linked to Tsw (0.9 +/- 0.6 cM) and is helpful for marker-assisted selection in a wide range of genetic intercrosses.

Alleles↗

Effect of cadmium on lipid composition of pepper.

Seedlings (2 weeks old) of pepper (Capsicum annum) were grown in nutrient solution with added CdCl(2) (10 or 50 microM) for 7 days. In Cd-treated plants, changes in acyl lipids and fatty acid composition were investigated. Cd particularly lowered the amount of monogalactosyldiacylglycerol (MGDG) and enhanced accumulation of phospholipids [phosphatidylcholine (PC), phosphatidylethanolamine and phosphatidylglycerol] in leaves. In contrast, content of PC and galactolipids (MGDG and digalactosyldiacylglycerol) decreased in roots. Fatty acid composition of leaves was also changed by Cd addition to external medium, but no important changes occurred in roots. Levels of leaf polyunsaturated fatty acids, especially 18:3 and 16:3, were reduced. Lipid and fatty acid composition changes in roots are discussed in relation to Cd tolerance in pepper.

Cadmium Chloride↗

A hot pepper cDNA encoding a pathogenesis-related protein 4 is induced during the resistance response to tobacco mosaic virus.

Hot pepper (Capsicum annuum) plants exhibit a hypersensitive response (HR) against infection by many tobamoviruses. A clone (CaPR-4) encoding a putative pathogenesis-related protein 4 was isolated by differential screening of a cDNA library prepared from resistant pepper plant leaves inoculated with tobacco mosaic virus (TMV) pathotype P0. The predicted amino acid sequence of CaPR-4 is very similar to those of other plant PR-4s. Southern blot analysis showed that small gene families of PR-4-related sequences were present in the pepper genome. Hot pepper cultivar Bugang, resistant to TMV-P0 and susceptible to TMV-P1.2, induced CaPR-4 expression by pathotype P0 inoculation in inoculated and systemic leaves, but not by pathotype P1.2. Effects of exogenously applied abiotic elicitors upon the CaPR-4 expression were also examined. The expression of the CaPR-4 gene was stimulated by methyl jasmonate (MeJA), ethephon and wounding treatment. However, application of salicylic acid (SA) did not trigger the expression. Evidence is emerging that jasmonic acid and ethylene play key roles in the SA-independent pathways of plant-pathogen interaction. Taken together, these results suggest that the CaPR-4 gene is one of the defense-related genes conferring resistance on pepper plants by the SA-independent pathway and the cross-talk between signaling compounds, jasmonic acid and ethylene could have a great regulatory potential in a plant's defense against TMV.

Acetates↗

Isolation of cDNA clones differentially accumulated in the placenta of pungent pepper by suppression subtractive hybridization.

Capsaicinoids responsible for pungency of chili pepper are synthesized exclusively in the placenta tissue of the fruit. As an elementary step in the molecular genetics study of capsaicinoid biosynthesis, a cDNA library was constructed from the placenta of a highly pungent pepper, Capsicum chinense cv. Habanero using the suppression subtractive hybridization (SSH). Thirty-nine cDNA clones from about 400 subtracted clones were selected through dot blot analysis and according to their nucleotides sequence. Sequence information of the chosen clones was evaluated by comparing it with DNA and protein databases. Results showed that the cDNA clones could be divided into 4 groups; cDNAs with similarities in genes encoding metabolic enzymes including acyl transferase and fatty acid alcohol oxidase (Group I), putative cell wall proteins (Group II), biotic and abiotic stress-inducible proteins (Group III), and lastly, cDNAs with no similarity (Group IV). Northern blot analysis was performed to confirm that these clones are differentially expressed in pungent pepper. The results revealed that all cDNA clones were differentially expressed in pungent pepper. In addition, the cDNA clones of Groups I and IV were differentially or preferentially expressed in the placenta of pungent pepper.

Blotting, Northern↗

Molecular cloning of the anther-preferential nonspecific lipid transfer protein cDNA in hot pepper by mRNA differential display.

To better understand the molecular control of anther development, an anther-preferential mRNA was isolated from hot pepper (Capsicum annuum) using mRNA differentially display. Using the displayed fragment as a probe, a full-length cDNA named CaLTP was isolated. A nucleotide sequence analysis of CaLTP revealed that the clone contains an open reading frame of 123 amino acids, which exhibits a 60-23% identity with nonspecific lipid transfer proteins (nsLTP). Northern and RT-PCR analysis of the clone confirmed that CaLTP mRNA was predominant to anther tissues. The basal expression level in the leaves was slightly induced only by abscisic acid (ABA) treatment. Southern analysis reveals that CaLTP is present as a single-copy gene in hot pepper genome. We hypothesize that CaLTP might have an important role in protecting the reproductive tissues from environmental stresses.

Amino Acid Sequence↗

Characterization of MADS box genes from hot pepper.

The cDNA clone, CanMADS1, was isolated from young flower buds of the hot pepper (Capsicum annuum L.) by screening a cDNA library using the OsMADS1 rice MADS-box gene as a probe. We used a yeast two-hybrid screening method to investigate interaction partners of the protein product of CanMADS1. A MADS-box gene, CanMADS6, was isolated from young flower buds using the region containing the K domain and 15 amino acid residues of the C-terminal region of CanMADS1 as a bait. CanMADS1 and CanMADS6 showed high amino acid sequence similarities to members of the AGL2 subfamily and the SQUA subfamily, respectively. CanMADS1 transcript was expressed in flower buds and fruits, and the transcription signal was the strongest in the stage of the fruit set (2 d after anthesis). CanMADS6 showed the same expression pattern as CanMADS1. CanMADS1 and CanMADS6 were not expressed in leaves. These results suggest that a regulatory role for flower and fruit development of the hot pepper may be accomplished through an interaction of the protein products of the two MADS-box genes, CanMADS1 and CanMADS6.

Amino Acid Sequence↗

Growth and photomorphogenesis of pepper plants under red light-emitting diodes with supplemental blue or far-red lighting.

Light-emitting diodes (LEDs) are a potential irradiation source for intensive plant culture systems and photobiological research. They have small size, low mass, a long functional life, and narrow spectral output. In this study, we measured the growth and dry matter partitioning of 'Hungarian Wax' pepper (Capsicum annuum L.) plants grown under red LEDs compared with similar plants grown under red LEDs with supplemental blue or far-red radiation or under broad spectrum metal halide (MH) lamps. Additionally, we describe the thermal and spectral characteristics of these sources. The LEDs used in this study had a narrow bandwidth at half peak height (25 nm) and a focused maximum spectral output at 660 nm for the red and 735 nm for the far-red. Near infrared radiation (800 to 3000 nm) was below detection and thermal infrared radiation (3000 to 50,000 nm) was lower in the LEDs compared to the MH source. Although the red to far-red ratio varied considerably, the calculated phytochrome photostationary state (phi) was only slightly different between the radiation sources. Plant biomass was reduced when peppers were grown under red LEDs in the absence of blue wavelengths compared to plants grown under supplemental blue fluorescent lamps or MH lamps. The addition of far-red radiation resulted in taller plants with greater stem mass than red LEDs alone. There were fewer leaves under red or red plus far-red radiation than with lamps producing blue wavelengths. These results indicate that red LEDs may be suitable, in proper combination with other wavelengths of light, for the culture of plants in tightly controlled environments such as space-based plant culture systems.

Biomass↗

Spectral quality affects disease development of three pathogens on hydroponically grown plants.

Plants were grown under light-emitting diode (LED) arrays with various spectra to determine the effects of light quality on the development of diseases caused by tomato mosaic virus (ToMV) on pepper (Capsicum annuum L.), powdery mildew [Sphaerotheca fuliginea (Schlectend:Fr.) Pollaci] on cucumber (Cucumis sativus L.), and bacterial wilt (Pseudomonas solanacearum Smith) on tomato (Lycopersicon esculentum Mill.). One LED (660) array supplied 99% red light at 660 nm (25 nm bandwidth at half-peak height) and 1% far-red light between 700 to 800 nm. A second LED (660/735) array supplied 83% red light at 660 nm and 17% far-red light at 735 nm (25 nm bandwidth at half-peak height). A third LED (660/BF) array supplied 98% red light at 660 nm, 1% blue light (BF) between 350 to 550 nm, and 1% far-red light between 700 to 800 nm. Control plants were grown under broad-spectrum metal halide (MH) lamps. Plants were grown at a mean photon flux (300 to 800 nm) of 330 micromoles m-2 s-1 under a 12-h day/night photoperiod. Spectral quality affected each pathosystem differently. In the ToMV/pepper pathosystem, disease symptoms developed slower and were less severe in plants grown under light sources that contained blue and UV-A wavelengths (MH and 660/BF treatments) compared to plants grown under light sources that lacked blue and UV-A wavelengths (660 and 660/735 LED arrays). In contrast, the number of colonies per leaf was highest and the mean colony diameters of S. fuliginea on cucumber plants were largest on leaves grown under the MH lamp (highest amount of blue and UV-A light) and least on leaves grown under the 660 LED array (no blue or UV-A light). The addition of far-red irradiation to the primary light source in the 660/735 LED array increased the colony counts per leaf in the S. fuliginea/cucumber pathosystem compared to the red-only (660) LED array. In the P. solanacearum/tomato pathosystem, disease symptoms were less severe in plants grown under the 660 LED array, but the effects of spectral quality on disease development when other wavelengths were included in the light source (MH-, 660/BF-, and 660/735-grown plants) were equivocal. These results demonstrate that spectral quality may be useful as a component of an integrated pest management program for future space-based controlled ecological life support systems.

Biomass↗

Construction and characterization of a bacterial artificial chromosome library from chili pepper.

A library of the bacterial artificial chromosome (BAC) that consisted of a total of 78,336 clones with an average insert size of 80 kb was constructed from Capsicum annuum, 'CM334', which is resistant to Phytophthora capsici and PVY. Based on a haploid genome size of pepper of 2,702 Mbp/C, the BAC library was estimated to contain approximately three genome equivalents and represented at least 90% of the pepper genome. In order to determine the percentage of BAC clones that contained mitochondrial DNAs, the entire library was screened with probes of chili pepper mitochondrial DNAs. The result showed that only twenty-five clones, which is 0.03% of the total BAC clones, were hybridized to mitochondrial gene probes. This indicates that the library is comprised predominantly of the nuclear sequences. The library was also tested for isolating specific clones by screening with a few known genes from the chili pepper, phytoene synthase gene, and two MADS genes--HpMADS1 and HpMADS3. The result showed that the three clones for phytoene synthase and the two clones for each MADS gene were positively hybridized to the specific probes. This indicates that the library is highly reliable and represents a resource for initiating map-based cloning and contig mapping in chili pepper.

Alkyl and Aryl Transferases↗

Weightlessness experiments on Biosatellite II.

Four experiments in the aft compartment of Biosatellite II investigated the broad question of the effect of nearly zero gravity on the development, morphology and metabolism of plants and animals. The fertilization and development of the egg of a vertebrate (the frog, Rana pipiens), the feeding and growth of a protozoan (the giant amoeba, Pelomyxa carolinensis), the orientation of leaves and petioles of a young dicotyledon (pepper plants, Capsicum annuum) and the morphogenesis, orientation, histochemistry and biochemistry of a monocotyledon seedling (wheat, Triticum vulgare) gave a broad scope. All are known to have specific responses to normal gravity and changes in them might be expected to reflect the effects of orbital flight on living organisms. No differences in development of the frog eggs could be detected. Unfortunately, the 3 1/2 hour delay in launch allowed the first cleavage (the stage most sensitive to inversion) to appear before launch. Although the orbited embryos were somewhat slower to reach certain stages of development, recovered embryos developed just as did the controls. The amoebae fed normally while in orbit, and specimens fixed in orbit retained the ordinary heteropodal shape. Growth rates of orbited amoebae, both fed and starved, were slower than controls following reentry and recovery procedures. In continuous-fed organisms there was little or no effect of flight detectable in growth rate or actual number of divisions. Electron micrographs showed no abnormalities and few differences between flight and control organisms. The pepper plants were photographed in orbit at ten-minute intervals, as were the clinostat and erect controls. The subsequent measurement of photographs showed that in the orbited plants all leaves showed epinasty, the interaxial angle decreasing by 20-60 degrees C. Plants on the horizontal clinostat behaved comparably, but recovered more rapidly than orbited plants when returned to the normal erect position. Although the maximum age of wheat seedlings was only 65 hours, coleoptile and root growth rates during that time had not been significantly altered by flight or by slow rotation on a horizontal clinostat. There was some evidence that growth was accelerated after normal gravity was restored. The orientation of coleoptiles and of primary and lateral roots of orbited plants varied significantly from the normal erect seedlings but was almost identical with that of clinostat plants. The Periodic-Acid-Schiff technique on sectioned material showed starch grains at the bottom of cells of erect control coleoptile and root tips, while in orbited and clinostated plants the grains were located more or less at random. Histochemical differences between clinostat and orbited tissues are apparent however. Peroxidase localization varied and its activity was higher in both clinostat and orbited tissues; five other enzymes studied biochemically showed no differences. These experiments all suggest that there is no deleterious effect on living organisms or their activities from short-term weightlessness. Several results indicate that the horizontal clinostat may simulate the weightless state effectively here on Earth.

Amoeba↗

[Effect of fertilization on the qualities of sweet pepper in greenhouse culture].

In order to study the effect of fertilization on the qualities of sweet pepper(Capsicum annul L) cultured in greenhouse, the experiment was carried out by drop irrigation. The results showed that the most sensitive organ of sweet pepper for nitrate was lateral branch, in which the nitrate content was highest. The nitrate content in leaf was lower than that in lateral branch, and in fruit, the nitrate content was the lowest. High fertilization of N, P and K had no negative influence on content of nitrate and total soluble sugars in fruit. The content of soluble sugars in fruit were about 50%-55% under all treatment and higher than that in leaf and lateral branch. Decreasing fertilization would cause increasing of the inosital and sucrose contents. Glucose and fructose were the main soluble sugars in fruit. The metabolism of carbohydrates was weakened by high fertilization, and it expressed as restraining the accumulation of starch content in leaf. The content of starch was the highest in fruit. The lipid content were exceed 3-4 times in the leaf than in the lateral branch and fruit, and perhaps, it was due to the higher content of chlorophyll in the leaf.

Capsicum↗