Detecting and estimating segregation distortion and linkage between glufosinate tolerance and blackleg resistance in Brassica napus L.

Summary The segregation and linkage between glufosinate (transgenes ‘Rf3’ and ‘T177’) and blackleg resistance genes in canola (Brassica napus L.) were assessed using F₁ microspore-derived doubled haploid (DH) populations from four crosses including reciprocals, two involving the transgene ‘Rf3’ and the other two involving the transgene ‘T177’. To relax the assumption of no segregation distortion required for the conventional analysis of segregation and linkage, we employed Bailey's analysis that allows detecting segregation distortion at linked loci. The significant departures from the 1:1 segregation were detected in the crosses involving the transgene ‘T177’ but not in the crosses involving the transgene ‘Rf3’. The apparent deficit of the herbicide tolerant DH lines in the crosses with the transgene ‘T177’ is likely due to differential selection against the gametes carrying ‘T177’ during microspore culture. The linkage was strong between blackleg resistance and the transgene ‘Rf3’ but weak or absent between blackleg resistance and the transgene ‘T177’, suggesting that the two transgenes are probably inserted into distant regions of the genome. The observed linkage offers an opportunity to develop new canola cultivars with both glufosinate tolerance conferred by transgene ‘Rf3’ and blackleg resistance.     

Histological and inheritance studies of partial resistance in the Brassica napus–Albugo candida host-pathogen interaction

Traditional and doubled haploid (DH) genotypes of oilseed Brassica spp. resistant, partially resistant, moderately susceptible, and susceptible to Albugo candida were compared for phenotypic development of host‐pathogen interaction and histology of host‐pathogen interaction. The partially resistant genotype showed pinhead‐size pustules, mainly on the upper surface of cotyledonary leaves. Relatively less mycelium was observed in the partially resistant genotype compared with the susceptible genotype. In resistant B. napus genotypes, there was neither pustule development nor any mycelial growth. In the moderately susceptible genotype, the pustules were similar to those in the partially resistant genotype in being of pinhead‐size and occasionally coalescing. However, ample mycelial growth in the mesophyll tissue in the moderately susceptible genotype was similar to that in the susceptible control B. rapa cv. ‘Torch’. The susceptible genotype B. rapa cv. ‘Torch’ also showed large coalescing pustules. In the non‐host B. juncea cv. ‘Commercial Brown’, no pustules were formed although some mycelial growth was observed beneath the epidermal cell layer and in the mesophyll cell layer of the cotyledonary leaf tissue. For inheritance studies, two partially resistant B. napus genotypes were crossed with a resistant B. napus genotype. Various generations viz., F1, F1(reciprocal), F₂, and DHs produced from the crosses were inoculated with a zoospore suspension of race 7v of A. candida. The partially resistant phenotype appeared to be controlled by a single recessive gene designated as wpr with variable expression. The simple inheritance of partial resistance has implications for disease resistance breeding against white rust, as this type of resistance can be easily incorporated into elite breeding lines through conventional and DH breeding methods.     

A Comparison of Genetic Segregation in Traditional and Microspore-Derived Populations of Brassica juncea L. Czern and Coss.

An isolated microspore culture procedure was used to produce doubled haploid lines of Brassica juncea from F¹ plants of reciprocal crosses between the cultivar‘RLM514’and a canola quality breeding line. The inheritance of two qualitative markers, seed color and leaf hairiness, was compared using traditional and microspore-derived populations from these crosses. Chi-square tests indicated that each trait is controlled by different sets of duplicate pairs of genes. Brown seeds or hairy leaves can result from the presence of either of two dominant alleles, whereas yellow seed or glabrous leaves are produced when alleles at both loci are recessive. The segregation of genes controlling seed color and leaf hairs in doubled haploid progeny did not differ significantly from that expected under random assortment, indicating that doubled haploids can be used in this species for genetic studies, and probably cultivar development as well.     

Selection for fast germination in rapeseed (Brassica napus L. and B. Campestris L.)

Summary Eleven populations of Brassica napus L. and twelve populations of B. campestris L. were subjected to three cycles of selection for fast germination at 2°C and at 25°C. The seeds from the selected populations, and unselected control populations grown in the same environment as the selected populations, were examined for germination behaviour at 2°C and 25°C, and for growth behaviour at 10°C. The populations in both species responded differently in terms of germination behaviour to selection for fast germination. In most of the populations that did respond positively to selection, selection practised at 2°C was superior to selection at 25°C in improving percent germination at 2°C, and was as good as the selection at 25°C in improving germination rate at the higher temperature. Selection for fast germination had no effect on growth characteristics of B. napus and B. campestris populations grown at 10°C. Thus, selection for fast germination at one low temperature may lead to improvement in germination characteristics over a range of temperatures, but will not necessarily lead to improved growth performance of the selected populations.     

Inheritance of partial resistance to race 2 of Albugo candida in canola-quality mustard (Brassica juncea) and its role in resistance breeding

The inheritance of partial resistance to race 2 of Albugo candida was studied in a canola-quality line of Brassica juncea . This partially resistant line was crossed with the susceptible B. juncea cultivar Commercial Brown. F₁, F₁(reciprocal), F₂, BC and doubled haploid generations from the cross were inoculated with a zoospore suspension of race 2 to study segregation of partial resistance. The partially resistant phenotype appeared to be controlled by a single dominant gene that has variable expression. This partial resistance can have implications in breeding for disease resistance against white rust, as adult plants did not develop hypertrophic growth or stagheads under greenhouse and field conditions.     

Exploitation of the late flowering species <Emphasis Type="Italic">Brassica oleracea</Emphasis> L. for the improvement of earliness in <Emphasis Type="Italic">B. napus</Emphasis> L.: an untraditional approach

The oilseed Brassica rapa flowers and matures earlier than B. oleracea, as well as their amphidiploid B. napus. Therefore, earliness of B. rapa has been investigated as a source of variation for earliness in B. napus breeding programs. Variation for days to flower exists in B. oleracea; however, its earliest flowering variant B. alboglabra flowers 2–3 weeks later than B. napus. We hypothesized that the C genome of B. alboglabra carries alleles for early flowering which are different from the C-genome alleles of B. napus; and these alleles can be used for the improvement of B. napus. To test this, we examined flowering time in pedigree and DH populations from two B. napus × B. alboglabra crosses. A B. napus line with about a week earlier flowering than the B. napus parent was achieved through reconstitution of its C genome following pedigree selection. Introgression of the B. alboglabra allele in the early flowering pedigree lines is also evident from the presence of B. alboglabra-specific SSR alleles in this line. However, application of doubled haploidy failed to generate any line that flowered earlier than the B. napus parent, which is probably due to the difficulty of obtaining large numbers of euploid B. napus DH lines from this interspecific cross. Thus, we demonstrate that a trait of the diploid species, which apparently looks undesirable, might in fact be highly valuable for the improvement of amphidiploids; and knowledge from this research can also be applied for other traits.     

Haploid plantlet screening in the development of blackleg resistant DH lines of Brassica napus

This study was conducted to expedite disease-resistance breeding in canola by screening haploid plants against blackleg disease. Microspore-derived haploid plantlets from 14 unrelated crosses were inoculated with Leptosphaeria maculans pycnidiospores and were rated as resistant, intermediate or susceptible. Blackleg-inoculated and control haploid plants were then colchicine-treated to produce doubled haploid (DH) lines. The DH lines thus produced were again screened against blackleg using the cotyledon bioassay. In general, the proportion of the resistant DH lines derived from selected resistant haploid plants was much higher than the proportion of resistant DH lines from control haploid populations. There was no detrimental effect on the survival of haploid plants after the combined treatment of blackleg inoculation and colchicine. The importance of the micro-environment on the final outcome of plant-pathogen interaction in case of non-obligate pathogens is also discussed.     

Interspecific cross of Brassica oleracea var. alboglabra and B. napus : effects of growth condition and silique age on the efficiency of hybrid production, and inheritance of erucic acid in the self-pollinated backcross generation

Interspecific hybrids were produced from reciprocal crosses between Brassica napus (2n = 38, AACC) and B. oleracea var. alboglabra (2n = 18, CC) to introgress the zero-erucic acid alleles from B. napus into B. oleracea. The ovule culture embryo rescue technique was applied for production of F₁ plants. The effects of silique age, as measured by days after pollination (DAP), and growth condition (temperature) on the efficiency of this technique was investigated. The greatest numbers of hybrids per pollination were produced under 20°/15°C (day/night) at 16 DAP for B. oleracea (♀) × B. napus crosses, while under 15°/10°C at 14 DAP for B. napus (♀) × B. oleracea crosses. Application of the ovule culture technique also increased the efficiency of BC₁ (F₁ × B. oleracea) hybrid production by 10-fold over in vivo seed set. The segregation of erucic acid alleles in the self-pollinated backcross generation, i.e. in BC₁S₁ seeds, revealed that the gametes of the F₁ and BC₁ plants carrying a greater number of A-genome chromosomes were more viable. This resulted in a significantly greater number of intermediate and a smaller number of high-erucic acid BC₁S₁ seeds.