New vegetable varieties of Brassica rapa and Brassica napus with modified glucosinolate content obtained by mass selection approach.

Background: Glucosinolates (GSLs) constitute a characteristic group of secondary metabolites present in the Brassica genus. These compounds confer resistance to pests and diseases. Moreover, they show allelopathic and anticarcinogenic effects. All those effects are dependent on the chemical structure of the GSL. The modification of the content of specific GSLs would allow obtaining varieties with enhanced resistance and/or improved health benefits. Moreover, the attainment of varieties with the same genetic background but with divergent GSLs concentration will prompt the undertaking of studies on their biological effects. Objective and Methods: The objective of this study was to evaluate the efficacy of two divergent mass selection programs to modify GSL content in the leaves of two Brassica species: nabicol (Brassica napus L.), selected by glucobrassicanapin (GBN), and nabiza (Brassica rapa L.), selected by gluconapin (GNA) through several selection cycles using cromatographic analysis. Results: The response to selection fitted a linear regression model with no signs of variability depletion for GSL modification in either direction, but with higher efficiency in reducing the selected GSL than in the increasing. The selection was also effective in other parts of the plant, suggesting that there is a GSL translocation in the plant or a modification in their synthesis pathway that is not-organ specific. There was an indirect response of selection in other GSL; thus this information should be considered when designing breeding programs. Finally, populations obtained by selection have the same agronomic performance or even better than the original population. Conclusion: Therefore, mass selection seems to be a good method to modify the content of specific GSL in Brassica crops.

Effects of 3-butenyl isothiocyanate on phenotypically different prostate cancer cells.

Isothiocyanates (ITCs) have gained increasing attention since they have been attributed the merits for the potential beneficial effects of cruciferous vegetable dietary consumption on cancer. The aim of the present study was to determine the cytotoxic effects of 3-butenyl ITC (3-BI) on prostate cancer (PC) cells under in vitro conditions. Two androgen-insensitive human PC cell lines, PC-3 and DU145, were assayed. Cells were cultured in the presence of increasing concentrations of 3-BI (5, 10, 30 and 50 µM) in the absence or presence of the chemotherapeutic drug docetaxel (DOCE) (1 and 2 nM). The cytotoxic effects of these compounds were analyzed using the trypan blue exclusion assay at 24, 48 and 72 h. Apoptosis and migration assays were also performed. The results showed that 3-BI induced a dose-dependent cytotoxic effect on PC-3 cells at 24, 48 and 72 h. These effects were significantly higher than those found with DOCE at 72 h of culture. Moreover, 3-BI also potentiated the effects of DOCE in a dose-dependent manner. Additionally, 3-BI showed inhibition of the migration of PC-3 cells. Nevertheless, 3-BI was not effective in the DU145 PC cell line. These results show a promising role for the 3-BI compound as a co-adjuvant agent in DOCE-based therapy in certain types of PC.

In vitro activity of glucosinolates and their degradation products against brassica-pathogenic bacteria and fungi.

Glucosinolates (GSLs) are secondary metabolites found in Brassica vegetables that confer on them resistance against pests and diseases. Both GSLs and glucosinolate hydrolysis products (GHPs) have shown positive effects in reducing soil pathogens. Information about their in vitro biocide effects is scarce, but previous studies have shown sinigrin GSLs and their associated allyl isothiocyanate (AITC) to be soil biocides. The objective of this work was to evaluate the biocide effects of 17 GSLs and GHPs and of leaf methanolic extracts of different GSL-enriched Brassica crops on suppressing in vitro growth of two bacterial (Xanthomonas campestris pv. campestris and Pseudomonas syringae pv. maculicola) and two fungal (Alternaria brassicae and Sclerotinia scletoriorum) Brassica pathogens. GSLs, GHPs, and methanolic leaf extracts inhibited the development of the pathogens tested compared to the control, and the effect was dose dependent. Furthermore, the biocide effects of the different compounds studied were dependent on the species and race of the pathogen. These results indicate that GSLs and their GHPs, as well as extracts of different Brassica species, have potential to inhibit pathogen growth and offer new opportunities to study the use of Brassica crops in biofumigation for the control of multiple diseases.

Intraspecific variation of host plant and locality influence the lepidopteran-parasitoid system of Brassica oleracea crops.

The aim of the study was to investigate the attractiveness to herbivores and parasitoids of two cultivars of Brassica oleracea L., namely, B. oleracea variety acephala (kale) and B. oleracea variety capitata (cabbage), that exhibit differences of morphological and biochemical traits. To this end, field samplings were replicated at seven localities in Galicia (northwestern Spain). Three specialist and three generalist lepidopteran species were sampled. In total, 7,050 parasitoids were obtained, belonging to 18 genera and 22 species. The results showed that 1) parasitism rate and parasitoid species richness changed with locality and was higher in cabbage, although this crop had lower herbivore abundance; 2) the proportion of specialist herbivores was higher in cabbage crops, whereas generalists dominated in kale crops; 3) the abundance of the parasitoids Telenomus sp. (Hymenoptera, Scelionidae), Cotesia glomerata L. (Hymenoptera: Braconidae), and Diadegma fenestrale (Holmgren) (Hymenoptera: Ichneumonidae) was higher in kale crops; and 4) parasitism rate of Pieris rapae larvae and pupae and Mamestra brassicae eggs were higher in kale crops. In contrast with the notion that plant structural complexity provides physical refuge to the hosts and can interfere with parasitoid foraging, parasitism rate was higher on cabbage plants, which form heads of overlapped leaves. Possibly, different chemical profiles of cultivars also influenced the host-parasitoid relationship. These results suggest that top-down and bottom-up forces may enhance cabbage crops to better control herbivore pressure during the studied season. In Spain, information on natural occurring parasitoid guilds of Brassica crops is still scarce. The data provided here also represent a critical first step for conservation biological control plans of these cultivations.

New insights into antioxidant activity of Brassica crops.

Antioxidant activity of six Brassica crops-broccoli, cabbage, cauliflower, kale, nabicol and tronchuda cabbage-was measured at four plant stages with DPPH and FRAP assays. Samples taken three months after sowing showed the highest antioxidant activity. Kale crop possessed the highest antioxidant activity at this plant stage and also at the adult plant stage, while cauliflower showed the highest antioxidant activity in sprouts and in leaves taken two months after sowing. Brassica by-products could be used as sources of products with high content of antioxidants. Phenolic content and composition varied, depending on the crop under study and on the plant stage; sprout samples were much higher in hydroxycinnamic acids than the rest of samples. Differences in antioxidant activity of Brassica crops were related to differences in total phenolic content but also to differences in phenolic composition for most samples.

Identification of Sources of Resistance to Xanthomonas campestris pv. campestris in Brassica napus Crops.

Black rot, caused by Xanthomonas campestris pv. campestris, is one of the most important diseases affecting Brassica crops worldwide. Nine races have been differentiated in X. campestris pv. campestris, with races 1 and 4 being the most virulent and widespread. The objective of this work was to identify sources of resistance to races 1 and 4 of X. campestris pv. campestris in different Brassica napus crops, mainly in the underexplored pabularia group. Seventy-six accessions belonging to four B. napus groups were screened for resistance to two X. campestris pv. campestris races (1 and 4). The strain of race 1 used in this study was more virulent on the tested materials than the strain of race 4. No race-specific resistance was found to race 1. Most cultivars were susceptible except Russian kale, from the pabularia group, which showed some resistant plants and some other accessions with some partially resistant plants. High levels of race-specific resistance to race 4 were found in the pabularia group, and great variability within accessions was identified. Three improved cultivars (Ragged Jack kale, Friese Gele, and Valle del Oro) and four landraces (Russian kale, MBG-BRS0037, MBG-BRS0041, and MBG-BRS0131) showed plants with some degree of resistance to both races, which may indicate that race-nonspecific resistance is involved. These accessions could be directly used in breeding programs, either as improved cultivars or as donors of race-specific resistance to other Brassica cultivars.

Resistance of cabbage (Brassica oleracea capitata group) crops to Mamestra brassicae.

Twenty-one cabbage (Brassica oleracea capitata group) varieties, including 16 local varieties and five commercial hybrids, were screened for resistance to the moth Mamestra brassicae L. under natural and artificial conditions in northwestern Spain. Resistance was assessed as the proportion of damaged plants and damaged leaves, leaf feeding injury, and number of larvae present. Correlation coefficients among damage traits showed that a visual scale (general appearance rating) should be a useful indicator of resistance. Most local varieties were highly susceptible to M. brassicae, whereas the commercial hybrids tested were resistant in terms of head foliage consumption and number of larvae per plant. Performance of varieties was similar under natural and artificial infestation although some of them performed differently at each year. Three local varieties (MBG-BRS0057, MBG-BRS0074, and MBG-BRS0452) were highly susceptible at both natural and artificial infestation conditions being MBG-BRS0074 the most damaged variety. Two local varieties (MBG-BRS0402 and MBG-BRS0535) and commercial hybrids were identified as resistant or moderately resistant to M. brassicae. Among them, 'Corazón de Buey' and 'Cabeza negra' were the most resistant and produced compact heads. These varieties could be useful sources of resistance to obtain resistant varieties to M. brassicae or as donors of resistance to other Brassica crops. The possible role of leaf traits, head compactness, and leaf glucosinolate content in relation to M. brassicae resistance is discussed.

Identification of quantitative trait loci for resistance to Xanthomonas campestris pv. campestris in Brassica rapa.

Resistance to six known races of black rot in crucifers caused by Xanthomonas campestris pv. campestris (Pammel) Dowson is absent or very rare in Brassica oleracea (C genome). However, race specific and broad-spectrum resistance (to type strains of all six races) does appear to occur frequently in other brassica genomes including B. rapa (A genome). Here, we report the genetics of broad spectrum resistance in the B. rapa Chinese cabbage accession B162, using QTL analysis of resistance to races 1 and 4 of the pathogen. A B. rapa linkage map comprising ten linkage groups (A01-A10) with a total map distance of 664 cM was produced, based on 223 AFLP bands and 23 microsatellites from a F(2) population of 114 plants derived from a cross between the B. rapa susceptible inbred line R-o-18 and B162. Interaction phenotypes of 125 F(2) plants were assessed using two criteria: the percentage of inoculation sites in which symptoms developed, and the severity of symptoms per plant. Resistance to both races was correlated and a cluster of highly significant QTL that explained 24-64% of the phenotypic variance was located on A06. Two additional QTLs for resistance to race 4 were found on A02 and A09. Markers closely linked to these QTL could assist in the transference of the resistance into different B. rapa cultivars or into B. oleracea.

Performance of crosses among flint maize populations under infestation by Sesamia nonagrioides (Lepidoptera: Noctuidae).

Flint maize, Zea mays L., varieties provide some interesting agronomic characteristics and kernels that possess a better ability than other kernels for developing high-quality flour. The pink stem borer, Sesamia nonagrioides Lefebvre, is an important constraint for the maize crop in Mediterranean regions. The objective of this work was to identify a "flint x flint" heterotic pattern that would perform well under artificial infestation by S. nonagrioides. A 10-population diallel was evaluated under infestation by S. nonagrioides in 2 yr. Variety effects were the only significant effects involved in stem and ear resistance to S. nonagrioides attack. Variety effects and average heterosis effects were the only significant effects for grain yield under artificial infestation conditions. Considering variety effects and cross-performance, the heterotic pattern Basto/Enano levantixo x Longfellow (BA/EL x LO) would be recommended for obtaining flint maize hybrids tolerant to S. nonagrioides attack because BA/EL had the most favorable variety effects for stem resistance, LO exhibited the most positive variety effects for grain yield, and the cross BA/EL x LO yielded significantly more than the remaining crosses.

Evaluation of the European Union Maize Landrace Core Collection for resistance to Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae).

Two corn borer species are the principal maize insect pests in Europe, the European corn borer, Ostrinia nubilalis (Hübner), and the pink stem borer, Sesamia nonagrioides (Lefebvre). Hence, it would be advisable to evaluate the European maize germplasm for corn borer resistance to generate European varieties resistant to corn borer attack. The creation of the European Union Maize Landrace Core Collection (EUMLCC) allowed the screening of most of the variability for European corn borer resistance present among European maize local populations from France, Germany, Greece, Italy, Portugal, and Spain, testing a representative sample. The objective of this study was the evaluation of stem and ear resistance of the EUMLCC to European corn borer and pink stem borer attack. Trials were made at two Spanish locations that represent two very different maize-growing areas. Populations that performed relatively well under corn borer infestation for stem and ear damage were 'PRT0010008' and'GRC0010085', among very early landraces; 'PRT00100120' and 'PRT00100186', among early landraces; 'GRC0010174', among midseason landraces; and 'ESP0070441', among late landraces. Either the selection that could have happen under high insect pressure or the singular origin of determined maize populations would be possible explanations for the higher corn borer resistance of some landraces. Landraces 'PRT0010008', 'FRA0410090', 'PRT00100186', and 'ESP0090214' would be selected to constitute a composite population resistant to corn borers and adapted to short season, whereas populations 'ESP0090033', 'PRT00100530', 'GRC0010174', and 'ITA0370005' would be used to make a resistant composite adapted to longer season.

Mean generation analysis of the damage caused by Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae) in sweet corn ears.

Sesamia nonagrioides (Lefebvre) and Ostrinia nubilalis (Hübner) are the main maize (Zea mays L.) pests in Mediterranean countries. To develop insect-resistant cultivars, it is helpful to know the genetic control of the resistance. Our objective was to determine the genetic control of the resistance to both borers. For each of two crosses (EP59 x P51 and 15125 x EP61), six generations were evaluated (P1, P2, F1, F2, BC1, and BC2). Genetic effects x environment interactions were not significant. For the O. nubilalis resistance traits; both crosses fitted an additive-dominance model. EP59 x P51 had important dominance and additive effects, whereas for 15125 x EP61 we did not detect significant genetic effects, but significant year effects were detected. For S. nonagrioides infestation, both crosses fitted to an additive-dominance model. There were additive effects for most traits in EP59 x P51. The cross I5125 x EP61 showed significant dominance effects for several traits. Resistance to both corn borers fit an additive-dominance model, but genetic effects depend on the cross evaluated. In the resistance to S. nonagrioides, additive effects were important for shank resistance, which is a useful trait for avoiding S. nonagrioides damage on the ear. Early sowings are recommended to make good use of the resistance to both corn borers. In the late sowings, damage is so high that resistant plants are not able to control the pest.

Resistance of kale populations to lepidopterous pests in northwestern Spain.

Kale (Brassica oleracea L. acephala) is common in northwestern Spain where it is severely damaged by different insect pests. Damage could be reduced by using resistant varieties. The objectives of this work were to evaluate the resistance of kale populations to leaf damage by lepidopterous pests, to determine which traits are the best indicators of resistance, and finally to study the relationship between the glossy phenotype and resistance. Fifteen kale populations, sowed early and late, were evaluated at two locations in northwestern Spain. Significant differences among genotypes were found for all damage traits. Damage was not related to planting dates. Highest levels of damage were observed from July to November. Some populations with different performance under natural infestation in 1999 were again evaluated in 2000 under artificial infestation with Mamestra brassicae (L.) eggs. Two accessions, MBC-BRS0142 and MBG-BRS0170, showed resistance to attack by lepidopterous pests. Correlation coefficients among damage traits show that general appearance rating may be an useful indicator of resistance. Phenotype of kale with glossy leaves seems to be related to resistance although further research is needed.

Antibiosis of the pith maize to Sesamia nonagrioides (Lepidoptera: Noctuidae).

Thirteen inbred lines of maize (Zea mays L.) with different levels of stem resistance to the stem borer Sesamia nonagrioides Lefevbre were evaluated in the field and the laboratory to determine the antibiotic resistance to this pest. Inbreds CM151, CO125, and EP39 had antibiotic pith as well as stem resistance, so the pith could play a role in stem resistance. Inbreds A509, F473, and PB130 did not have antibiotic pith but had stem resistance; therefore, other mechanisms could confer stem resistance. Finally, the inbred MS1334 had antibiotic pith and did not show stem resistance; thus, other factors could compensate the effect of the pith. Therefore, although pith antibiotic compound seems to play a role in the defense against S. nonagrioides attack, it is not the only possible mechanism of defense.