Sequence Comparison of Plant Ornithine Decarboxylases Reveals High Homology and Lack of Introns

We have designed and constructed four oligonucleotides corresponding to the most conserved regions of ornithine decarboxylases (ODC; EC 4.1.1.17) of plant origin. These oligonucleotides were used for the amplification of homologous fragments from several plants (Zea mays, Capsicum annuum, Sorghum bicolor, Phaseolus vulgaris, Carica papaya and Daucus carota). The amplified fragments were cloned and sequenced, revealing high homology to other ODCs. Peptide sequences coded by these fragments were compared by Clustal analyses. These analyses identified the location of the conserved sequences corresponding to the binding sites of substrate and cofactor. Data demonstrated that the plant ODCs fragments lacked intron sequences and were extremely homologous (over 80 %), constituting a compact group separated from other eukaryotic ODCs. This revised version was published online in July 2006 with corrections to the Cover Date.     

Broccoli growth parameters and level of head infestations in simple and mixed plantings: Impact of increased flora diversification

The impacts of interplanting broccoli, Brassica oleracea var. italica L., with chili pepper, Capsicum annuum L., or yellow sweetclover (YSC), Melilotus officinalis L., on broccoli growth parameters and levels of lepidopteran head infestations were studied during two field seasons (winter & summer). During both seasons, several broccoli growth characteristics (e.g. plant vertical area, whole plant biomass, etc.) were significantly reduced in broccoli‐YSC habitats. However, broccoli heads in broccoli‐YSC habitats did not differ significantly in size from other treatment habitats. Additionally, these heads contained the lowest mean lepidopteran contaminants per broccoli crown during the winter season. During this period, the average number of lepidopteran contaminants per broccoli head was more than twice that in monoculture and pepper intercropped broccoli than in broccoli‐YSC habitats. During the summer experiment, the average number of lepidopteran contaminants per broccoli head was greater in pepper intercropped broccoli than in both monoculture and YSC interplanted broccoli. Potential causes of broccoli growth differences and their relationship to lepidopteran ovipositional behavior are discussed.     

Marker-assisted introgression of 4 Phytophthora capsici resistance QTL alleles into a bell pepper line: validation of additive and epistatic effects

The aim of the present study was to transfer resistance to P. capsici alleles at four quantitative trait loci (QTLs) from a small fruited pepper into a bell pepper recipient line using markers. The marker-assisted selection program was initiated from a doubled-haploid line issued from the mapping population and involved three cycles of marker-assisted backcross (MAB). Two populations, derived by selfing the plants selected after the first selection cycle, were genotyped and evaluated phenotypically for their resistance level. The additive and epistatic effects of the four resistance factors were re-detected and validated in these populations, indicating that introgression of 4 QTLs in this MAB program was successful. A decrease of the effect for the moderate-effect QTLs and of the epistatic interaction was observed. Phenotypic evaluations of horticultural traits were performed on sample of each backcross generation. The results indicated an efficient return to the recipient phenotype using this MAB strategy.     

Seed Storage Proteins in Solanaceae and Cucurbitaceae Species

Electrophoretic analyses of non-reduced and reduced seed storage proteins from Solanaceae and Cucurbitaceae species and cultivars were performed. High molecular disulfide bonded complexes between intermediary subunits of 11S globulins previously detected in Capsicum annuum cultivars, were found in Solanum melongena cultivars as well. The data obtained might be used for further elucidation of peculiarities of the 11S globulins in dicotyledonous plants.     

Exposure Time to Lethal Temperatures for Meloidogyne incognita Suppression and Its Implication for Soil Solarization

Meloidogyne incognita eggs or J2 were incubated in test tubes containing sand:peat mix and immersed in a water bath heated to 38, 39, 40, 41, 42, 43, 44 and 45°C for a series of time intervals. Controls were maintained at 22°C. Nematodes surviving or hatching were collected from Baermann trays after three weeks of incubation. Regression analyses between percent survival or egg hatch and hours of heat treatment were performed for each temperature. Complete suppression of egg hatch required 389.8, 164.5, 32.9, 19.7 and 13.1 hours at 38, 39, 40, 41 and 42°C, respectively. Complete killing of J2 required 47.9, 46.2, 17.5 and 13.8 hours at 39, 40, 41 and 42°C, respectively. J2 were not completely killed at 38°C within 40 hours of treatment, but were killed within one hour at 44 and 45°C. Effect of temperature on nematode killing is not determined by heat units. Oscillating temperature between cool and warm did not interfere with the nematode suppressive effect by the heat treatment. Six-week solarization in the field during the summers of 2003 and 2004 in Florida accumulated heat exposure times in the top 15 cm of soil that surpassed levels required to kill M. incognita as determined in the water bath experiments. Although near zero M. incognita were detected right after solarization, the nematode population densities increased after a cycle of a susceptible pepper crop. Therefore, future research should address failure of solarization to kill nematodes in the deeper soil layers.     

Cytogenetic markers for the characterization of Capsicum annuum L. cultivars

Karyotypic studies with conventional staining have been unsuccessful due to the uniformity of Capsicum chromosomes. In this study, we found diagnostic chromosome characters that permit to characterize cultivars; this is the first cytological characterization of both rDNAs (18S and 5S) in a species of Capsicum using a genus-specific probe and the most exhaustive in C. annuum to date. The heterochromatic banding patterns enabled us to identify cultivars, and fluorescent in situ hybridization (FISH) showed one 5S rDNA locus largely conserved within the cultivars, whereas high variation in the number of 18S rDNA loci was observed. One of the most obvious differences is the presence of an additional active nucleolar organizer region in pair #12 and the dispersal of inactive 18S rDNA signals. These results indicate that fluorochrome banding together with silver impregnation and FISH procedures are very useful for the identification of chromosomes and the interpretation of chromosomal variation between cultivars. The functional role of this variation is still uncertain, but our results show that copy number variation of repetitive DNA during the course of evolution might provide an excellent experimental system for studying genome rearrangements accompanying functional divergence in domesticated C. annuum.     

Isolation and culture of protoplasts ofCapsicum annuum L. and their regeneration into plants floweringin vitro

Summary Axenic shoot cultures ofCapsicum annuum cv.California Wonder were used as the source for isolation of protoplasts from mesophyll cells. Protoplasts underwent sustained mitotic activity and proliferated to form callus masses on NT or DPD medium enriched with 2,4-D, NAA and BAP each at 1 mg/l level. The callus could be differentiated into whole plants on the differentiation media and plants floweredin vitro under long day conditions.     

Effects of compactin,a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor,on the growth of alfalfa (Medicago sativa) seedlings and the rhizogenesis of pepper (Capsicum annuum) explants

The effects of compactin, a specific inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, on the growth of alfalfa seedlings in vivo and the rhizogenesis of pepper explants in vitro were investigated. Compactin added to the agar medium inhibited the elongation of roots and hypocotyls of etiolated alfalfa seedlings. The growth inhibition was accompanied by strict inhibition of sterol synthesis. Addition of mevalonic acid, the direct product of 3-hydroxy-3-methylglutaryl coenzyme A reductase, together with compactin relieved the growth inhibition. The sterol level in the seedlings was also protected against the lowering effect of compactin. Similarly, the rhizogenetic process of cultured explants of pepper was inhibited by compactin and relieved by mevalonic acid. Several isoprenoid end products were tested in combination with compactin to determine which compounds, if any, might be limiting for growth. Exogenously supplied isoprenoids failed to relieve the growth inhibition of seedlings. In contrast, they partly relieved the growth inhibition of explants, suggesting their important role in plant growth. During the course of these experiments, it was also found that brassinolide caused remarkable growth inhibition and twisting of alfalfa seedlings.