Oleracea Cultivars Research Articles

Microsatellite-based analysis of genetic diversity in 91 commercial Brassica oleracea L. cultivars belonging to six varietal groups

Brassica oleracea L. includes various types of important vegetables that show extremely diverse phenotypes. To elucidate the genetic diversity and relationships among commercial cultivars derived by different companies throughout the world, we characterized the diversity and genetic structure of 91 commercial B. oleracea cultivars belonging to six varietal groups, including cabbage, broccoli, cauliflower, kohlrabi, kale and kai-lan. We used 69 polymorphic microsatellite markers showing a total of 359 alleles with an average number of 5.20 alleles per locus. Polymorphism information content (PIC) values ranged from 0.06 to 0.73, with an average of 0.40. Among the six varietal groups, kohlrabi cultivars exhibited the highest heterozygosity level, whereas kale cultivars showed the lowest. Based on genetic similarity values, an UPGMA clustering dendrogram and a two-dimensional scale diagram (PCoA) were generated to analyze genetic diversity. The cultivars were clearly separated into six different clusters with a tendency to cluster into varietal groups. Model-based structure analysis revealed six genetic groups, in which cabbage cultivars were divided into two subgroups that were differentiated by their head shape, whereas cauliflower and kai-lan cultivars clustered together into a single group. Furthermore, we identified 18 SSR markers showing 27 unique alleles specific to only one cultivar that can be used to discriminate 22 cultivars from the others. Our phylogenetic and population structure analysis presents new insights into the genetic structure and relationships among 91 B. oleracea cultivars and provides valuable information for breeding of B. oleracea species. In addition, we demonstrate the utility of SSR markers as a powerful tool for discriminating between the cultivars. The SSR markers described herein will also be helpful for Distinctness, Uniformity and Stability (DUS) test of new cultivars.

Phloem‐specific resistance in Brassica oleracea against the whitefly Aleyrodes proletella

AbstractThe cabbage whitefly [Aleyrodes proletella L. (Hemiptera: Aleyrodidae)] is becoming a serious pest in Brassica oleracea L. (Brassicaceae) crops. However, almost nothing is known about the interaction of this insect with its host plants. Previous studies have shown differences in the natural occurrence of adults, eggs, and nymphs on the closely related B. oleracea cultivars Christmas Drumhead and Riviera grown in the field. In this study, we aimed to identify the nature of these differences and to gain insight into the resistance mechanisms against A. proletella. We used no‐choice experiments on field‐ and greenhouse‐grown plants to show that the differences between the two cultivars are mainly based on antibiosis (traits that reduce herbivore performance) and not on antixenosis (traits that deter herbivory). This was further supported by laboratory choice experiments that indicated little or no discrimination between the two cultivars based on plant volatiles. We showed that resistance is dependent on plant age, that is, resistance increased during plant development, and is mainly independent of environmental factors. Analysis of probing behaviour revealed that the resistance trait affects A. proletella at the phloem level and that morphological differences between the two cultivars are most likely not involved. We suggest that compounds present in the phloem reduce sap ingestion by the whitefly and that this explains the observed resistance.

Relative importance of plant-mediated bottom-up and top-down forces on herbivore abundance on Brassica oleracea

1. Arthropod communities are structured by complex interactions between bottom-up (resource-based) and top-down (natural enemy-based) forces. Their relative importance in shaping arthropod communities, however, continues to be under debate. Bottom-up and top-down forces can be affected by intraspecific plant variation, for example by differences in concentrations of secondary metabolites that affect herbivore abundance through plant quality (bottom-up) or attract natural enemies of these herbivores (top-down). 2. Our objective was to investigate whether herbivore abundance is more strongly affected by plant-mediated bottom-up or top-down forces. 3. We used a model system of four cultivars of Brassica oleracea that show a high degree of variation in several plant traits, resistance to herbivores and attraction of natural enemies. During two field seasons, we recorded the abundance of several herbivorous and carnivorous insect species. To assess the relative importance of bottom-up and top-down forces, we quantified chemical and morphological traits of the cultivars (bottom-up) and assessed parasitization of herbivores and predator oviposition on plants inoculated with a controlled number of herbivores (top-down). 4. We show that intraspecific variation in plant chemistry and morphology consistently affects the abundance of insect herbivores and their natural enemies, resulting in cascading effects on tritrophic interactions in the associated insect community. Foliar profiles of glucosinolates and leaf toughness appeared most important for these effects. Brassica oleracea cultivars that harboured the largest numbers of herbivores also harboured the largest numbers of natural enemies. Differences in the fraction of herbivores parasitized and in predator oviposition on plants inoculated with a controlled number of herbivores could not explain the differences in natural abundance of herbivores. 5. Although abundance of herbivores is most likely influenced by a combination of bottom-up and top-down forces, it appears that in the tritrophic system investigated, bottom-up forces (plant chemistry and morphology) were more important for herbivore abundance than plant-mediated top-down forces (attraction and arrestment of natural enemies).

Effects of host plant and genetic background on the fitness costs of resistance to Bacillus thuringiensis

Novel resistance to pathogens and pesticides is commonly associated with a fitness cost. However, measurements of the fitness costs of insecticide resistance have used diverse methods to control for genetic background and rarely assess the effects of environmental variation. Here, we explored how genetic background interacts with resource quality to affect the expression of the fitness costs associated with resistance. We used a serially backcrossed line of the diamondback moth, Plutella xylostella, resistant to the biopesticide Bacillus thuringiensis, to estimate the costs of resistance for insects feeding on two Brassica species. We found that fitness costs increased on the better-defended Brassica oleracea cultivars. These data were included in two meta-analyses of fitness cost experiments that used standardized protocols (and a common resistant insect stock) but which varied in the methodology used to control for the effects of genetic background. The meta-analysis confirmed that fitness costs were higher on the low-quality host (B. oleracea); and experimental methodology did not influence estimates of fitness costs on that plant species. In contrast, fitness costs were heterogeneous in the Brassica pekinensis studies: fitness costs in genetically homogenized lines were significantly higher than in studies using revertant insects. We hypothesize that fitness modifiers can moderate fitness costs on high-quality plants but may not affect fitness when resource quality is low.

Combined effects of patch size and plant nutritional quality on local densities of insect herbivores

Plant–insect interactions occur in spatially heterogeneous habitats. Understanding how such interactions shape density distributions of herbivores requires knowledge on how variation in plant traits (e.g. nutritional quality) affects herbivore abundance through, for example, affecting movement rates and aggregation behaviour. We studied the effects of plant patch size and herbivore-induced differences in plant nutritional quality on local densities of insect herbivores for two Brassica oleracea cultivars, i.e. white cabbage and Brussels sprouts. Early season herbivory as a treatment resulted in measurable differences in glucosinolate concentrations in both cultivars throughout the season. Herbivore induction and patch size both influenced community composition of herbivores in both cultivars, but the effects differed between species. Flea beetles (Phyllotreta spp.) were more abundant in large than in small patches, and this patch response was more pronounced on white cabbage than on Brussels sprouts. Herbivore-induction increased densities in all patches. Thrips tabaci was also more abundant in large patches and densities of this species were higher on Brussels sprouts than on white cabbage. Thrips densities were lower on induced than on control plants of both cultivars and this negative effect of induction tended to be more pronounced in large than in small patches. Densities of the cabbage moth (Mamestra brassicae) were lower on Brussels sprouts than on white cabbage and lower on herbivore-induced than on uninduced plants, with no effect of patch size. No clear effects of patch size and induction were found for aphids. This study shows that constitutive and herbivore-induced differences in plant traits interact with patch responses of insect herbivores.

Intraspecific variation in herbivore community composition and transcriptional profiles in field-grown Brassica oleracea cultivars.

Intraspecific differences in plant defence traits are often correlated with variation in transcriptional profiles and can affect the composition of herbivore communities on field-grown plants. However, most studies on transcriptional profiling of plant–herbivore interactions have been carried out under controlled conditions in the laboratory or greenhouse and only a few examine intraspecific transcriptional variation. Here, intraspecific variation in herbivore community composition and transcriptional profiles between two Brassica oleracea cultivars grown in the field is addressed. Early in the season, no differences in community composition were found for naturally occurring herbivores, whereas cultivars differed greatly in abundance, species richness, and herbivore community later in the season. Genome-wide transcriptomic analysis using an Arabidopsis thaliana oligonucleotide microarray showed clear differences for the expression levels of 26 genes between the two cultivars later in the season. Several defence-related genes showed higher levels of expression in the cultivar that harboured the lowest numbers of herbivores. Our study shows that herbivore community composition develops differentially throughout the season on the two B. oleracea cultivars grown in the field. The correlation between the differences in herbivore communities and differential expression of particular defence-related genes is discussed.

Field parasitism rates of caterpillars on Brassica oleracea plants are reliably predicted by differential attraction of Cotesia parasitoids

Summary 1. Herbivore‐induced plant volatiles (HIPVs) play an important role in host location of parasitoid wasps and may benefit the plant by top–down control of its herbivorous attackers. Although many studies have shown that accessions of plants differ in attractiveness to parasitoid wasps under controlled laboratory studies, few studies have confirmed that the most attractive accessions also sustain highest parasitism rates in the field. Here, we tested whether in‐flight preference of parasitoids for HIPVs from cultivars of Brassica oleracea in the laboratory reliably predicts the parasitism rates of herbivores feeding on these cultivars in the field. 2. In wind tunnel tests in the laboratory, we ranked cultivars of B. oleracea for the preference of two congeneric parasitoids (Cotesia glomerata and C. rubecula) for their HIPVs. The cultivars were then compared for their relative parasitism rates of caterpillars in the field. Throughout the growth season in the field, we infested the different cultivars with Pieris caterpillars on a weekly basis. The caterpillars were recollected after 3 days, dissected and scored for the rate of parasitism. 3. Cultivars of B. oleracea that we identified as most attractive to parasitoids in the laboratory also sustained highest proportions of parasitism in the field. The composition of the headspace of the B. oleracea cultivars damaged by P. rapae differs among these cultivars in the amounts of terpenoids and methyl salicylate emitted, which may be responsible for the differential attraction of parasitoids to the cultivars. 4. Our results show that intraspecific variation in HIPVs of plants is paralleled by differential parasitism of caterpillars in the field. The widely used laboratory assays on HIPV‐based preferences of parasitoids provided reliable information on relative parasitism differences of herbivores as found in the field. 5. Thereby, our work confirms that through HIPVs plants attract parasitoids that effectively parasitize herbivores even under the complex and variable abiotic and biotic conditions in (agro‐) ecosystems.

Chemical diversity in Brassica oleracea affects biodiversity of insect herbivores

Intraspecific variation in plants plays a major role in the composition and diversity of the associated insect community. Resistance traits of plants are likely candidates mediating community composition. However, it is debated whether total concentrations of chemical compounds or specific compounds determine herbivore resistance, and how chemical diversity among plant genotypes in turn affects the composition of the associated herbivore community. To study the role of specific chemical compounds in affecting the herbivore community, we used cultivated Brassica oleracea. The cultivars differ qualitatively in glucosinolate profile, i.e., foliar composition of different glucosinolate compounds, and only a little in total concentration of glucosinolates, the secondary metabolites specific for the Brassicaceae family. In field and laboratory experiments, we tested whether individual compounds explained differences in herbivore community composition, and whether herbivores with a similar degree of host plant specialization responded in a similar way to variation in glucosinolate profiles. In the field B. oleracea cultivars differed widely in species richness and composition of the herbivore community, as well as in the density of insects they harbored. Plants with high concentrations of the short side chain alkenyl glucosinolate, glucoiberin, harbored low herbivore diversity. Higher biodiversity was found when plants had glucosinolate profiles containing high concentrations of glucosinolates with elongated side chains, which are biosynthetically linked to glucoiberin. Although glucosinolates are known to have differential effects on generalist and specialist herbivores, all herbivore species exhibited similar responses to the intraspecific variation in foliar glucosinolate profiles of the B. oleracea cultivars. This observation is supported by the correspondence between oviposition preferences of the specialist herbivore Pieris rapae and the generalist Mamestra brassicae in the field and the laboratory, using the same cultivars, and may be due to the relatively low concentrations of glucosinolates in cultivars. Our results show that variation in the concentration of short side-chain glucosinolates affects the composition of the herbivore community associated with brassicaceous plants.

Responses of Brassica oleracea cultivars to infestation by the aphid Brevicoryne brassicae: an ecological and molecular approach

Intraspecific variation in resistance or susceptibility to herbivorous insects has been widely studied through bioassays. However, few studies have combined this with a full transcriptomic analysis. Here, we take such an approach to study the interaction between the aphid Brevicoryne brassicae and four white cabbage (Brassica oleracea var. capitata) cultivars. Both under glasshouse and field conditions, two of the cultivars clearly supported a faster aphid population development than the other two, indicating that aphid population development was largely independent of the environmental conditions. Genome-wide transcriptomic analysis using 70-mer oligonucleotide microarrays based on the Arabidopsis thaliana genome showed that only a small number of genes were differentially regulated, and that this regulation was highly cultivar specific. The temporal pattern in the expression behaviour of two B. brassicae-responsive genes in all four cultivars together with targeted studies employing A. thaliana knockout mutants revealed a possible role for a trypsin-and-protease inhibitor in defence against B. brassicae. Conversely, a xyloglucan endotransglucosylase seemed to have no effect on aphid performance. Overall, this study shows clear intraspecific variation in B. brassicae susceptibility among B. oleracea cultivars under glasshouse and field conditions that can be partly explained by certain differences in induced transcriptional changes.

Performance of specialist and generalist herbivores feeding on cabbage cultivars is not explained by glucosinolate profiles

AbstractPlants display a wide range of chemical defences that may differ in effectiveness against generalist and specialist insect herbivores. Host plant‐specific secondary chemicals such as glucosinolates (GS) in Brassicaceae typically reduce the performance of generalist herbivores, whereas specialists have adaptations to detoxify these compounds. The concentration of glucosinolates may also alter upon herbivory, allowing the plant to tailor its response to specifically affect the performance of the attacking herbivore. We studied the performance of three Lepidoptera species, two specialists [Pieris rapae L. (Pieridae), Plutella xylostella L. (Yponomeutidae)] and one generalist [Mamestra brassicae L. (Noctuidae)], when feeding on eight cultivars of Brassica oleracea L. and a native congener (Brassica nigra L.) and related this to the GS content. We tested the hypotheses (i) that a generalist herbivore is more affected by high GS concentrations, and (ii) that generalist feeding has a stronger effect on GS levels. Although performance of the three herbivores was different on the B. oleracea cultivars, M. brassicae and P. xylostella had a similar ranking order of performance on the eight cultivars. In most of the cultivars, the concentration of indole GS was significantly higher after feeding by P. rapae or M. brassicae than after P. xylostella feeding. As a consequence, the total concentration of GS in the cultivars showed a different ranking order for each herbivore species. The generalist M. brassicae performed equally well as the specialist P. xylostella on cultivars with high concentrations of GS. Our findings suggest that secondary metabolites other than GSs or differences in nutrient levels affect performance of the species studied.

Development and reproductive potential of diamondback moth Plutella xylostella (Lepidoptera: Plutellidae) on cultivated and wild crucifer species in Kenya

The development, survival and reproductive potential of diamondback moth Plutella xylostella (Linnaeus) were studied at 25 ± 1 °C in the laboratory in response to two cultivated Brassica oleracea cultivars (cabbage B. oleracea var. capitata and kale B. oleracea var. acephala) and four wild crucifer species Erucastrum arabicum, Raphanus raphanistrum, Rorippa nudiuscula and Rorippa micrantha. Rorippa micrantha was the most preferred species in oviposition choice tests, while cabbage and kale were least preferred. First instar larval mining period differed significantly between plant species with the longest period recorded on cabbage (3.0 days) and the shortest on R. micrantha (0.4 days). Pupal weight was significantly lower for larvae reared on R. nudiuscula, while those of the others were similar. The developmental period from first instar to adult was the shortest on R. micrantha (14.1 days) and the longest on R. raphanistrum (15.6 days). Survival to adult was not statistically affected by the host plant species. Adult longevity ranged between 18.2 days on R. raphanistrum and 24.7 days on R. nudiuscula. The females were significantly heavier than the males on all plant species. However, males lived longer than females. Moths reared on R. nudiuscula recorded the highest fecundity (326 eggs), while moths reared on cabbage had the lowest fecundity (262 eggs). Kale and R. nudiuscula recorded the longest generation time of 31.7 days, while E. arabicum had the highest net reproductive rate (126.4 eggs per day). The highest intrinsic rate of increase was calculated for R. micrantha (0.179) and the lowest for kale (0.147). This study shows the suitability of wild crucifers as hosts for P. xylostella and indicates that they may play a major role as reservoir for the pest during the absence of cultivated host plants.

Database derived microsatellite markers (SSRs) for cultivar differentiation in Brassica oleracea

Fifty-nine Brassica oleracea cultivars, belonging to five botanical varieties, were evaluated for microsatellite (SSR) polymorphisms using 11 database sequence derived primer pairs. The cultivars represented 12 broccoli (Brassica oleracea var. italica), ten Brussels sprouts (B. o. var. gemmifera), 21 cabbage (B. o. var. capitata, including the groups white and red cabbage), six savoy cabbage (B. o. var. sabauda), and ten cauliflower (B. o. var. botrytis) cultivars from 13 seed suppliers. The 11 primer pairs amplified in total 47 fragments, and differentiated 51 of the cultivars, whereas the remaining eight cultivars were differentiated from the rest in four inseparable pairs. All SSR markers, except one, produced a polymorphic information content (PIC value) of 0.5 or above. The average diversity for all markers within the tested material was 0.64. There was no major difference in the diversity within botanical varieties and groups. The cluster analysis and the resulting dendrogram showed that the cultivars tended to group within these taxonomic units. The present study substantiates the use of microsatellite markers as a powerful tool for cultivar differentiation and identification in vegetable brassicas.

Linkage mapping of a dominant male sterility geneMs-cd1inBrassica oleracea

The dominant male sterility gene Ms-cd1 (c, cabbage; d, dominant) was identified as a spontaneous mutation in the spring cabbage line 79-399-3. The Ms-cd1 gene is successfully applied in hybrid seed production of several Brassica oleracea cultivars in China. Amplified fragment length polymorphism (AFLP) technology was used to identify markers linked to the Ms-cd1 gene in bulks of male-sterile and male-fertile individuals of a segregating BC3 population and in a near-isogenic population of 25 male-sterile plants. Twelve markers within a 20-cM interval proximal to the Ms-cd1 gene were identified, 5 of which can be used to select homozygous male-sterile Ms-cd1/ Ms-cd1 plants. Three AFLP markers and 3 sequence characterized amplified region markers that were linked to MS-cd1 mapped onto linkage group O9, corresponding to chromosome 3 of B. oleracea. This region corresponds to the top of chromosome 5 in Arabidopsis thaliana.

Transfer of Powdery Mildew Resistance from Brassica carinata to B. oleracea

Powdery mildew is a fungal disease of crucifers, caused by Erysiphe polygoni D.C. and it can be problematic during seed increase in green-houses. Crosses were made between Brassica carinata (Ethiopian mustard) accession (PI 360883) and B. oleracea cultivars `Titleist' and `Cecile' to transfer resistance to powdery mildew to B. oleracea germplasm. It was not possible to obtain interspecific hybrids between Ethiopian mustard and B. oleracea through natural seed set. However, interspecific hybrids and backcross one (BC1) progenies were produced via embryo rescue following sexual crosses. Four interspecific hybrid plants were produced with the aid of embryo rescue from cultured pistils with B. carinata as the maternal parent, and their interspecific origin was confirmed through plant morphology and analysis of RAPD polymorphisms. No interspecific hybrids were obtained when `Titleist' was used as a maternal parent. Interspecific hybrid plants were male sterile and they were used as maternal parents to produce BC1 plants. Twenty one BC1 plants were obtained through natural seed set and embryo rescue, although embryo rescue was not necessary to produce first backcross generation plants. When tested in greenhouse with powdery mildew, all interspecific hybrids and eight of the BC1 plants were resistant to the disease. Crosses are being made to produce BC2 plants with 2n = 18 chromosomes for introgression of the resistance in B. oleracea.

Genetic relationships of Brassica vegetables determined using database derived simple sequence repeats

Sequence databases were screened to identify simple sequence repeats (SSRs) in Brassica oleracea sequences. A total of 512 B. oleracea DNA sequences were screened and 43 potential SSRs were identified. Thirty-six primer pairs were designed to amplify target sequences. Of the 36 primer pairs, six failed to amplify fragments of expected sizes, and 17 primer pairs failed to generate polymorphisms. Thirteen SSRs were used to assess genetic similarity between 54 B. oleracea cultivars, belonging to 3 variteal groups (cabbage, cauliflower, and broccoli). Pairwise genetic similarities were calculated for cultivars, and a dendrogram of relationships was produced. All cabbage cultivars were distinguished from each other and clustered in two separate groups. Five cauliflower cultivars could not be distinguished with SSR markers used in the study. Three broccoli cultivars clustered with cauliflower cultivars, and two cauliflower cultivars grouped with broccoli cultivars. The varietal group with the narrowest genetic variation in the study was cauliflower (B. oleracea var. botrytis) followed by broccoli (B. oleracea var. italica) and cabbage (B. oleracea var. capitata) groups. Polymorphism information content (PIC) values and number of alleles produced per marker ranged between 0.25 to 0.86 and 1 to 8, respectively, for database derived SSR markers.

Development of black rot resistant interspecific hybrids between Brassica oleracea L. cultivars and Brassica accession A 19182, using embryo rescue

Black rot is a bacterial disease of Brassica oleracea caused by Xanthomonas campestris pv. campestris. Resistance to the major black rot races 1 or 4 has been identified in related Brassica species including B. carinata and B. napus. In this study, two B. juncea accessions (A 19182 and A 19183) that are resistant to races 1 and 4 of Xcc were used as maternal and paternal parents to generate interspecific hybrids with B. oleracea cultivars. Interspecific hybrids were recovered using the embryo rescue technique and confirmed through inheritance of paternal molecular markers. Twenty-six interspecific hybrid plants were obtained between A 19182 and B. oleracea cultivars, but no interspecific hybrids were obtained using A 19183. Although interspecific hybrid plants were male sterile, they were used successfully as maternal parents to generate backcross plants using embryo rescue. All hybrid and BC1 plants were resistant to black rot races 1 and 4.

Internal transcribed spacer 2 amplicon as a molecular marker for identification of Peronospora parasitica (crucifer downy mildew).

The purpose of the study was to characterize the internal transcribed spacer (ITS) regions of Peronospora parasitica (crucifer downy mildew) in order to evaluate their potential as molecular markers for pathogen identification. PCR amplification of ribosomal RNA gene block (rDNA) spacers (ITS1 and ITS2) performed in 44 P. parasitica isolates from different Brassica oleracea cultivars and distinct geographic origins, revealed no length polymorphisms. ITS restriction analysis with three endonucleases, confirmed by sequencing, showed no fragment length polymorphisms among isolates. Furthermore, ITS amplification with DNA isolated from infected host tissues also allowed the detection of the fungus in incompatible interactions. The combination of the universal ITS4 and ITS5 primers, for amplification of full ITS, with a new specific forward internal primer for ITS2 (PpITS2F), originates a P. parasitica specific amplicon, suitable for diagnosis. As ITS2 regions of P. parasitica, B. oleracea, other B. oleracea fungal pathogens and other Peronospora species are clearly distinct, a fast and reliable molecular identification method based on multiplex PCR amplification of full ITS and P. parasitica ITS2 is proposed for the diagnosis of crucifer downy mildew. The method can be applied to diagnose the disease in the absence of fungal reproductive structures, thus being useful to detect nonsporulating interactions, early stages of infection on seedlings, and infected young leaves packed in sealed plastic bags. Screening of seed stocks in sanitary control is also a major application of this diagnostic method.

The effects of anther culture and plant genetic background on Agrobacterium rhizogenes-mediated transformation of commercial cultivars and derived doubled-haploid Brassica oleracea

The production of transgenic roots was scored for eight Brassica oleracea cultivars from broccoli, cabbage, cauliflower and kale following inoculation with an Agrobacterium rhizogenes cell line carrying a binary plasmid bearing the green fluorescence protein (gfp) gene in the T-DNA. Significant differences in the numbers of explants producing transgenic roots were observed between cultivars, ranging from 1.4% for Marathon F1 to 57.8% for the Green Duke F1. Three F1 cultivars were subjected to anther culture, and doubled-haploid (DH) lines were used for transformation. The DH lines produced showed considerable variation for transgenic root production with some lines showing increased efficiency compared to the parental F1 cultivar. Grouping of the DH lines into response classes with respect to transgenic root production allowed the development of potential genetic models to explain the variation in performance released from each F1 cultivar. No apparent segregation distortion for transgenic root production was observed in the DH lines following anther culture.

S-HAPLOTYPE POLYMORPHISM IN BRASSICA OLERACEA

Self-incompatibility (SI) promotes outcrossing in Brassica species and has been exploited for producing Fl hybrids in cole crops. The SI locus (S-locus) is very polymorphic and cross-compatibility between plants depends on the allelic forms (S-haplotypes) of genes at the S-locus. Among these genes, SLG (S-Locus Glycoprotein gene) is specifically expressed in the stigma. Two classes (class I and class II) of SLG genes have been described and antibodies allowing a specific identification of class I or class II SLG are available. In this paper, we report the polymorphism in Brassica oleracea cultivars (cauliflower, broccoli, romanesco, cabbage, kale and wild forms of the Normandy coast) as assessed by using an isoelectric focusing and immunostaining combined analysis. Plants from an S-allele collection obtained from Dr D. Ockendon were also introduced in the analysis to allow the identification of S-haplotypes according to his nomenclature. Using the two class-specific-SLG antibodies, approximately 80 different immunostaining patterns were identified out of the 950 plants analysed. However, we were unable to find specific immunostaining patterns for about 10% of the analysed plants. Among these plants, we identified 9 S-haplotypes by crossing experiments with lines of known S-haplotype. We also observed that three S-haplotypes (S2, S29, S63) gave variable immunostaining patterns, where SLG bands were either clearly detectable, faintly detectable or not present. This indicates that, in these particular S-haplotypes, the amount of SLG is highly dependent on the genetic background of the plants. All but one of the S-' haplotypes described in the wild forms were also found in cultivated forms. The three class II haplotypes (i.e., S2, S5, S15) were detected in wild Brassica but only S15 was common to all cultivated as well as wild Brassica plants. Moreover, S15 was the most frequent S-haplotype detected in the species.

Diamondback Moth (Lepidoptera: Plutellidae) Resistance of Brassica napus and B. oleracea Lines with Differing Leaf Characteristics

Canola quality oil produced from oilseed rape, Brassica napus L. (Brassicaceae), is a relatively new crop in the United States and is damaged by a number of insect pests, including the crucifer specialist diamondback moth, Plutella xylostella (L.). Greenhouse tests were performed to evaluate resistance in normal-leafed ('Bingo', 'Cobra', 'Oscar') and glossy-leafed B. napus plants ('glossy Bingo' and plant introductions (PI) 469895, 470055, 470055-2, and 470064) to identify potential resistant sources to diamondback moth. Normal-leafed PI 171538, originally described as B. napus and recently reclassified as B. oleracea, also was evaluated for resistance with B. napus plants and B. oleracea cultivars of broccoli, 'Green Comet', 'Green Valiant', and collard 'Vates'. Plants of Cornell line 92-8611, a broccoli with glossy-leaf type (glossy broccoli), were used as the resistant control in all trials. B. napus entries failed to show feeding nonpreference or antibiosis comparable to that of glossy broccoli, Furthermore, no significant differences were detected between normal and glossy-leafed B. napus lines for most of the parameters tested. Within the tested B. napus entries, PI 470055 exhibited low levels of feeding damage in the restricted-feeding test and some level of antibiosis against diamondback moth. However, B. oleracea entry PI 171538 showed a greater or comparable level of resistance to that of glossy broccoli. Nonpreference for larval feeding and adult oviposition were exhibited by PI 171538. In feeding tests, PI 171538 was less defoliated compared with other B. oleracea entries. Diamondback moth larvae reared on PI 171538 leaves for 5 d had a lower head capsule width compared with larvae reared on other entries. In an oviposition preference test, PI 171538 received 0.9% of eggs as compared with normal-leafed B. napus plants (2.1-4.4%) and glossy broccoli, which received 7.0% of eggs. The experiments indicate that the normal-leafed B. oleracea entry PI 171538 has considerable resistance against diamondback moth. PI171538 is a newly discovered source of resistance and has leaf characteristics that differ from the previously reported diamondback moth-resistant B. oleracea lines.