Neil Coombes

Genome-wide delineation of natural variation for pod shatter resistance in Brassica napus

Resistance to pod shattering (shatter resistance) is a target trait for global rapeseed (canola, Brassica napus L.), improvement programs to minimise grain loss in the mature standing crop, and during windrowing and mechanical harvest. We describe the genetic basis of natural variation for shatter resistance in B. napus and show that several quantitative trait loci (QTL) control this trait. To identify loci underlying shatter resistance, we used a novel genotyping-by-sequencing approach DArT-Seq. QTL analysis detected a total of 12 significant QTL on chromosomes A03, A07, A09, C03, C04, C06, and C08; which jointly account for approximately 57% of the genotypic variation in shatter resistance. Through Genome-Wide Association Studies, we show that a large number of loci, including those that are involved in shattering in Arabidopsis, account for variation in shatter resistance in diverse B. napus germplasm. Our results indicate that genetic diversity for shatter resistance genes in B. napus is limited; many of the genes that might control this trait were not included during the natural creation of this species, or were not retained during the domestication and selection process. We speculate that valuable diversity for this trait was lost during the natural creation of B. napus. To improve shatter resistance, breeders will need to target the introduction of useful alleles especially from genotypes of other related species of Brassica, such as those that we have identified.

Evaluating landraces of bread wheat Triticum aestivum L. for tolerance to aluminium under low pH conditions

Bread wheat (Triticum aestivum L.) landraces held within ex situ collections offer a valuable and largely unexplored genetic resource for wheat improvement programs. To maximise full utilisation of such collections the evaluation of landrace accessions for traits of interest is required. In this study, 250 accessions from 21 countries were screened sequentially for tolerance to aluminium (Al) using haematoxylin staining of root tips and by root regrowth measurement. The staining test indicated tolerance in 35 accessions, with an intermediate response to Al exhibited in a further 21 accessions. Of the 35 accessions classified as tolerant, 33 also exhibited increased root length following exposure to Al. The tolerant genotypes originated from Bulgaria, Croatia, India, Italy, Nepal, Spain, Tunisia, and Turkey. AFLP analysis of the 35 tolerant accessions indicated that these represent diverse genetic backgrounds. These accessions form a valuable set of germplasm for the study of Al tolerance and may be of benefit to breeding programs for expanding the diversity of the gene pool from which tolerant cultivars are developed.

Genome‐wide association analyses reveal complex genetic architecture underlying natural variation for flowering time in canola

Optimum flowering time is the key to maximize canola production in order to meet global demand of vegetable oil, biodiesel and canola-meal. We reveal extensive variation in flowering time across diverse genotypes of canola under field, glasshouse and controlled environmental conditions. We conduct a genome-wide association study and identify 69 single nucleotide polymorphism (SNP) markers associated with flowering time, which are repeatedly detected across experiments. Several associated SNPs occur in clusters across the canola genome; seven of them were detected within 20 Kb regions of a priori candidate genes; FLOWERING LOCUS T, FRUITFUL, FLOWERING LOCUS C, CONSTANS, FRIGIDA, PHYTOCHROME B and an additional five SNPs were localized within 14 Kb of a previously identified quantitative trait loci for flowering time. Expression analyses showed that among FLC paralogs, BnFLC.A2 accounts for ~23% of natural variation in diverse accessions. Genome-wide association analysis for FLC expression levels mapped not only BnFLC.C2 but also other loci that contribute to variation in FLC expression. In addition to revealing the complex genetic architecture of flowering time variation, we demonstrate that the identified SNPs can be modelled to predict flowering time in diverse canola germplasm accurately and hence are suitable for genomic selection of adaptative traits in canola improvement programmes.

Influence of plant silicon and sugarcane cultivar on mandibular wear in the stalk borer Eldana saccharina.

1 Silicon can increase the resistance of plants to attack by herbivorous insects. The present study aimed to determine the effect of silicon and cultivar on mandibular wear in larvae of the sugarcane stalk borer Eldana saccharina Walker (Lepidoptera: Pyralidae).

Sponge and dough bread making: genetic and phenotypic relationships with wheat quality traits

The genetic and phenotypic relationships among wheat quality predictors and sponge and dough bread making were evaluated in a population derived from a cross between an Australian cultivar ‘Chara’ and a Canadian cultivar ‘Glenlea’. The genetic correlation across sites for sponge and dough loaf volume was high; however, phenotypic correlations across sites for loaf volume were relatively low compared with rheological tests. The large difference between sites was most likely due to temperature differences during grain development reflected in a decrease in the percentage of unextractable polymeric protein and mixing time. Predictive tests (mixograph, extensograph, protein content and composition, micro-zeleny and flour viscosity) showed inconsistent and generally poor correlations with end-product performance (baking volume and slice area) at both sites, with no single parameter being effective as a predictor of end-product performance. The difference in the relationships between genetic and phenotypic correlations highlights the requirement to develop alternative methods of selection for breeders and bakers in order to maximise both genetic gain and predictive assessment of grain quality.

Genome-wide Association Study Identifies New Loci for Resistance to Leptosphaeria maculans in Canola

Key message “We identified both quantitative and quantitative resistance loci to Leptosphaeria maculans, a fungal pathogen, causing blackleg disease in canola. Several genome-wide significant associations were detected at known and new loci for blackleg resistance. We further validated statistically significant associations in four genetic mapping populations, demonstrating that GWAS marker loci are indeed associated with resistance to L. maculans. One of the novel loci identified for the first time, Rlm12, conveys adult plant resistance in canola.” Blackleg, caused by Leptosphaeria maculans, is a significant disease which affects the sustainable production of canola (Brassica napus). This study reports a genome-wide association study based on 18,804 polymorphic SNPs to identify loci associated with qualitative and quantitative resistance to L. maculans. Genomic regions delimited with 694 significant SNP markers, that are associated with resistance evaluated using 12 single spore isolates and pathotypes from four canola stubble were identified. Several significant associations were detected at known disease resistance loci including in the vicinity of recently cloned Rlm2/LepR3 genes, and at new loci on chromosomes A01/C01, A02/C02, A03/C03, A05/C05, A06, A08, and A09. In addition, we validated statistically significant associations on A01, A07, and A10 in four genetic mapping populations, demonstrating that GWAS marker loci are indeed associated with resistance to L. maculans. One of the novel loci identified for the first time, Rlm12, conveys adult plant resistance and mapped within 13.2 kb from Arabidopsis R gene of TIR-NBS class. We showed that resistance loci are located in the vicinity of R genes of Arabidopsis thaliana and Brassica napus on the sequenced genome of B. napus cv. Darmor-bzh. Significantly associated SNP markers provide a valuable tool to enrich germplasm for favorable alleles in order to improve the level of resistance to L. maculans in canola.

Molecular mapping and validation of Rlm1 gene for resistance to Leptosphaeria maculans in canola (Brassica napus L.)

Abstract. European winter canola (Brassica napus L.) cultivars harbour genes for durable resistance to the fungus Leptosphaeria maculans, which causes blackleg disease under Australian environmental conditions. Previous studies have shown that resistance in winter-type cultivars Maxol and Columbus is controlled by two genes, Rlm1 and Rlm3, which have been mapped using randomly amplified polymorphic DNA markers onto chromosome A7. We mapped a doubled-haploid population that consisted of 101 lines from a cross between Maxol*1 and Westar-10 using diversity arrays technology and simple sequence repeat (SSR)-based markers. Two SSR marker loci, Xol12-e03 and Xra2-a05b, flanked the Rlm1 locus at an interval of 6.7 cM, which corresponds to ∼3.2 Mb of the Brassica rapa genomic sequence; this region contains several genes encoding putative kinase and leucine-rich repeat-type disease-resistance proteins. SSR markers were further tested for their linkage with the Rlm1 locus in an independent population derived from Columbus*3/Westar-10. Our results showed that SSR markers linked to Rlm1 can be useful for monitoring Rlm1 gene introgression in breeding populations derived from Maxol and Columbus.

Effect of fipronil seed treatments on the germination and early growth of rice

Fipronil seed treatments were evaluated to determine whether they directly influence germination and subsequent seedling growth in rice (Oryza sativa L). Continuous seed exposure to fipronil (four days) at 2000 mg litre -1 significantly impaired germination (P 0.05) were found between shoot lengths or root system dry weights of control plants and plants developing from seed exposed continuously (two days) to fipronil at rates of up to 2000 mg litre -1 during germination and harvested nine days after sowing. Treating germinated seed with fipronil for 1 h immediately prior to sowing at rates of up to 4000 mg litre -1 did not result in significant changes (P<0.05) in plant growth parameters at either nine or 25 days after sowing. No evidence of fipronil having a direct phytostimulatory effect on rice was obtained.

Genetic improvement of triticale for irrigated systems in south-eastern Australia: a study of genotype and genotype × environment interactions

Abstract. Research into winter cereal breeding in Australia has focused primarily on studying the effects of rainfed environments. These studies typically show large genotype × environment (GE) interactions, and the complexity of these interactions acts as an impediment to the efficient selection of improved varieties. Wheat has been studied extensively; however, there are no published studies on the GE interactions of triticale in Australia under irrigated production systems. We conducted trials on 101 triticale genotypes at two locations over 4 years under intensive irrigated management practices and measured the yield potential, GE interactions, heritability and estimated genetic gain of yield, lodging resistance and several other traits important for breeding triticale. We found that high yield potential exceeding 10 t ha–1 exists in the Australian germplasm tested and that, in these irrigated trials, genotype accounted for a high proportion of the variability in all measured traits. All genetic parameters such as heritability and estimated genetic gain were high compared with rainfed studies. Breeding of triticale with improved yield and lodging resistance for irrigated environments is achievable and can be pursued with confidence in breeding programs.

Molecular diversity analysis and genetic mapping of pod shatter resistance loci in Brassica carinata L.

Seed lost due to easy pod dehiscence at maturity (pod shatter) is a major problem in several members of Brassicaceae family. We investigated the level of pod shatter resistance in Ethiopian mustard (Brassica carinata) and identified quantitative trait loci (QTL) for targeted introgression of this trait in Ethiopian mustard and its close relatives of the genus Brassica. A set of 83 accessions of B. carinata, collected from the Australian Grains Genebank, was evaluated for pod shatter resistance based on pod rupture energy (RE). In comparison to B. napus (RE = 2.16 mJ), B. carinata accessions had higher RE values (2.53 to 20.82 mJ). A genetic linkage map of an F2 population from two contrasting B. carinata selections, BC73526 (shatter resistant with high RE) and BC73524 (shatter prone with low RE) comprising 300 individuals, was constructed using a set of 6,464 high quality DArTseq markers and subsequently used for QTL analysis. Genetic analysis of the F2 and F2:3 derived lines revealed five statistically significant QTL (LOD ≥ 3) that are linked with pod shatter resistance on chromosomes B1, B3, B8, and C5. Herein, we report for the first time, identification of genetic loci associated with pod shatter resistance in B. carinata. These characterized accessions would be useful in Brassica breeding programs for introgression of pod shatter resistance alleles in to elite breeding lines. Molecular markers would assist marker-assisted selection for tracing the introgression of resistant alleles. Our results suggest that the value of the germplasm collections can be harnessed through genetic and genomics tools.