Important Brassica Species Research Articles

Sequence-level comparative analysis of theBrassica napusgenome around two stearoyl-ACP desaturase loci

We conducted a sequence-level comparative analyses, at the scale of complete bacterial artificial chromosome (BAC) clones, between the genome of the most economically important Brassica species, Brassica napus (oilseed rape), and those of Brassica rapa, the genome of which is currently being sequenced, and Arabidopsis thaliana. We constructed a new B. napus BAC library and identified and sequenced clones that contain homoeologous regions of the genome including stearoyl-ACP desaturase-encoding genes. We sequenced the orthologous region of the genome of B. rapa and conducted comparative analyses between the Brassica sequences and those of the orthologous region of the genome of A. thaliana. The proportion of genes conserved (approximately 56%) is lower than has been reported previously between A. thaliana and Brassica (approximately 66%). The gene models for sets of conserved genes were used to determine the extent of nucleotide conservation of coding regions. This was found to be 84.2 +/- 3.9% and 85.8 +/- 3.7% between the B. napus A and C genomes, respectively, and that of A. thaliana, which is consistent with previous results for other Brassica species, and 97.5 +/- 3.1% between the B. napus A genome and B. rapa, and 93.1 +/- 4.9% between the B. napus C genome and B. rapa. The divergence of the B. napus genes from the A genome and the B. rapa genes was greater than anticipated and indicates that the A genome ancestor of the B. napus cultivar studied was relatively distantly related to the cultivar of B. rapa selected for genome sequencing.

Genetics of Clubroot Resistance in Brassica Species

Clubroot disease, caused by the obligate plant pathogen Plasmodiophora brassicae Wor., is one of the most economically important diseases affecting Brassica crops in the world. The genetic basis of clubroot resistance (CR) has been well studied in three economically important Brassica species: B. rapa, B. oleracea, and B. napus. In B. rapa, mainly in Chinese cabbage, one minor and seven major CR genes introduced from European fodder turnips have been identified. Mapping of these CR genes localized Crr1 on R8, Crr2 on R1, CRc on R2, and Crr4 on R6 linkage groups of Chinese cabbage. Genes Crr3, CRa, CRb, and CRk mapped to R3, but at two separate loci, CRa and CRb are independent of Crr3 and CRk, which are closely linked. Further analysis suggested that Crr1, Crr2, and CRb have similar origins in the ancestral genome as in chromosome 4 of Arabidopsis thaliana. Genetic analysis of clubroot resistance genes in B. oleracea suggests that they are quantitative traits. Twenty-two quantitative trait loci (QTLs) were mapped in different linkage groups of B. oleracea. In B. napus, genetic analysis of clubroot resistance was found to be governed by one or two dominant genes, whereas resistance conferred by two recessive genes is reported. The quantitative analysis approach, however, proved that they are polygenic. In total, at least 16 QTLs have been detected on eight chromosomes of B. napus, N02, N03, N08, N09, N13, N15, N16, and N19. The chromosomal location of the other six QTLs is not clear. Cloning of any of these QTLs or resistance loci was not, however, possible until recently. Progress in genomics, particularly the techniques of comparative mapping and genome sequencing, supplements cloning and allows improved characterization of CR genes. Further development of DNA markers linked to CR genes will in turn hasten the breeding of clubroot-resistant Brassica cultivars.

Genetic Variation in Carotenoid Concentrations among Diploid and Amphidiploid Rapid-cycling Brassica Species

Vegetable crops can be significant sources of nutritionally important dietary carotenoids, and Brassica are sources that also exhibit antioxidant and anticarcinogenic activity. The family Brassicaceae contains a diverse group of plant species commercially important in many parts of the world. The six economically important Brassica species are closely related genetically. Three diploid species (B. nigra, B. rapa, B. oleracea) are the natural progenitors of the amphidiploid species (B. juncea, B. napus, B. carinata). The objective of this study was to characterize the accumulation of important dietary carotenoid pigments among the genetically related Brassica species. High-performance liquid chromatographic quantification revealed significant differences in carotenoid and chlorophyll pigment concentrations among the Brassica species. Brassica rapa accumulated the highest concentrations of antheraxanthin [0.79 mg/100 g fresh weight (FW)], lutein (8.89 mg/100 g FW), and zeaxanthin (0.75 mg/100 g FW). The highest concentrations of β-carotene (4.41 mg/100 g FW) and total chlorophyll (125.9 mg/100 g FW) were found in B. juncea. Brassica nigra accumulated the highest concentrations of 5,6-epoxylutein (0.41 mg/100 g FW) and violaxanthin (2.28 mg/100 g FW), whereas B. oleracea accumulated the highest concentrations of neoxanthin (2.10 mg/100 g FW). For many of the pigments analyzed, the amphidiploids B. carinata and B. napus accumulated significantly less carotenoid concentrations than the diploid species and B. juneca. Brassica convey unique health attributes when consumed in the diet. Identification of genetic relationships among the Brassica species would be beneficial information for improvement programs designed to increase carotenoid values.

HPLC screening of anti-cancer sulforaphane from important European Brassica species

Natural sulforaphane has been of increasing interest for nutraceutical and pharmaceutical industries due to its anti-cancer effect. The sulforaphane contents of 14 different major European Brassica oleracea L. varieties (cauliflower Alverda, cauliflower di Jesi, cauliflower Minaret, broccoli Ramoso calabrese, white cabbage, palm cabbage Cuor di Bue, broccoli Primor, savoy cabbage riccio d’Asti, broccoli Eolo, Delicato, cauliflower Nuvola, cauliflower Palla di neve, San martino, Velox) and four different Brassica rapa L. varieties (turnip Senza Testa, cima di rapa 40, cima di rapa 90, cima di rapa 120) which are cultivated in Italy, were analysed during two different developmental stages (3rd and 9th day after germination) of the seedlings. A quantitative determination of sulforaphane in Brassica species seedlings was established by HPLC. The highest amount of sulforaphane (2.21 mg/g d w) was found in San martino 3-day old seedlings. Seedlings of this European Brassica variety may be considered as a potential new source of natural sulforaphane.

(167) Carotenoid Accumulation Among the Diploid and Amphidiploid Brassica Species

Vegetable crops can be significant sources of nutritionally important dietary carotenoids and Brassica vegetables are sources that also exhibit antioxidant and anticarcinogenic activity. The family Brassicaceae contains a diverse group of plant species commercially important in many parts of the world. The six economically important Brassica species are closely related genetically. Three diploid species (B. nigra, B. rapa, and B. oleracea) are the natural progenitors of the allotetraploid species (B. juncea, B. napus, and B. carinata). The objective of this study was to characterize the accumulation of important dietary carotenoid pigments among the genetically related Brassica species. The HPLC quantification revealed significant differences in carotenoid and chlorophyll pigment accumulation among the Brassica species. Brassica nigra accumulated the highest concentrations of lutein, 5,6-epoxy lutein, violaxanthin, and neoxanthin. The highest concentrations of beta-carotene and total chlorophyll were found in B. juncea. Brassica rapa accumulated the highest concentrations of zeaxanthin and antheraxanthin. For each of the pigments analyzed, the diploid Brassica species accumulated higher concentrations, on average, than the amphidiploid species. Brassicas convey unique health attributes when consumed in the diet. Identification of genetic relationships among the Brassica species would be beneficial information for improvement programs designed to increase carotenoid values.

Identification of Alternaria brassicicola genes expressed in planta during pathogenesis of Arabidopsis thaliana

Alternaria brassicicola is a necrotrophic fungal pathogen that causes black spot disease on cruciferous plants including economically important Brassica species. The purpose of this study was to identify fungal genes expressed during infection of Arabidopsis. In order to identify candidate genes involved in pathogenicity, we employed suppression subtractive hybridization (SSH) between RNA isolated from A. brassicicola spores incubated in water and on the leaf surface of the Arabidopsis ecotype Landsberg. Two populations of cDNA were created from total RNA extracted after 24 h when approximately 80% of the spores had germinated either on the leaf surface or in water. Following SSH, expression of clones was examined using dot-blot macro-arrays and virtual Northern blots. 47 cDNA clones differentially expressed between Alternaria infected Arabidopsis leaves and spore germination in water were selected for sequencing. Seventy-seven percent (36) of the cDNAs had significant homology to fungal sequences from databases examined, including available fungal genomes, while 13% (11) had no homology to sequences in the databases. All 36 genes had significant matches with genes of fungal origin, while 11 genes did not have significant hits in the databases examined. Five sequences were expressed on the plant leaf surface but not during spore germination in water according to virtual Northern blots. These five cDNAs were predicted to encode a cyanide hydratase, arsenic ATPase, formate dehydrogenase, major Alternaria allergen, and one unknown. RT-PCR was used to examine the expression of these five genes during infection of Brassica oleraceae var. capitata (cabbage), in vitro growth in nutrient rich media, and infection of Arabidopsis thaliana. Four of these genes are expressed in the nutrient rich medium, while the unknown gene P3F2 was only expressed during plant infection. The results of this study provide the first insight into genes expressed during A. brassicicola infection of Brassica species that may be involved in fungal pathogenesis.

Realizing the potential of rapid-cycling Brassica as a model system for use in plant biology research.

Rapid-cycling Brassica populations were initially developed as a model for probing the genetic basis of plant disease. Paul Williams and co-workers selected accessions of the six main species for short time to flower and rapid seed maturation. Over multiple generations of breeding and selection, rapid-cycling populations of each of the six species were developed. Because of their close relationship with economically important Brassica species, rapid-cycling Brassica populations, especially those of B. rapa (RCBr) and B. oleracea, have seen wide application in plant and crop physiology investigations. Adding to the popularity of these small, short-lived plants for research applications is their extensive use in K-12 education and outreach.

Physical mapping of rDNA loci in Brassica species

The number of major rDNA loci (the genes coding for 18S-5.8S-26S rRNA) was investigated in the economically important Brassica species and their wild relatives by in situ hybridization of an rDNA probe to metaphase chromosomes and interphase nuclei. The diploid species B. nigra (B genome) has two major pairs of rDNA loci, B. oleracea (C genome) has two major pairs and one minor pair of loci, while B. campestris (A genome) has five pairs of loci. Among the three tetraploid species arising from these three diploid ancestors, B. carinata (BBCC genomes) has four loci, B. juncea (AABB genomes) has five major pairs and one minor pair of loci, and B. napus (AACC genomes) has six pairs of loci, indicating that the number of loci has been reduced during evolution. The complexity of the known rDNA restriction fragment length polymorphism patterns gave little indication of number of rDNA loci. It is probable that chromosome rearrangements have occurred during evolution of the amphidiploid species. The data will be useful for physical mapping of genes relative to rDNA loci, micro- and macro-evolutionary studies and analysis of aneuploids including addition and substitution lines used in Brassica breeding programs.

Protoplast culture and plant regeneration of different agronomically important Brassica species and varieties

Protoplast cultures were prepared from 6-day-old hypocotyls of six spring, seven winter cultivars of Brassica napus L. and one line of Brassica campestris L. The molarity of enzyme solution was raised to 0,714 M mannitol resulting in well manipulable, cytoplasm dense protoplasts. In the protoplast purification procedure density gradient centrifugation was used to minimize physical damage of protoplasts. Three different protoplast culture systems —(1) liquid, (2) 2nd day embedded, (3) directly embedded in low melting agarose were compared. The two different protoplast embedding techniques resulted in the same efficiency of cell division as the liquid culture method and over this fact the colony browning was avoided. Using protoplast agarose-embedding and culture techniques, healthy calli were obtained for plant regeneration experiments. Incorporation of silver nitrate into the regeneration medium improved the efficiency of plant regeneration in responsive genotypes and the regeneration was induced in three nonresponsive (without silver nitrate) genotypes, too. The supplement of silver nitrate in regeneration medium was especially advantageous in plant regeneration of B. campestris. Out of fourteen commercial cultivars of Brassica napus and B. campestris, there is only one recalcitrant genotype in obtaining plantlets from protoplast-derived calli.