Identification of boron tolerance in Brassica rapa

There has been increasing interest in developing canola quality B. juncea for low rainfall areas across Australia over the past two decades. However, B. juncea genotypes are susceptible to high levels of boron in Western Victorian soils. An understanding of the genetics and the molecular basis of boron tolerance may enable fast and accurate tolerance selection and lead to improved boron tolerance. Being an allotetraploid species, B. juncea is difficult to understand at the genetic level because of chromosomal duplication and the potential presence of multiple copies of the loci of interest. Therefore, once the tolerance genes or chromosomal loci governing tolerance are identified in the diploid progenitor genomes, B. rapa and B. nigra, boron tolerant B. juncea lines may be resynthesized. Thus, as an initial step in this process, this thesis aimed to understand the physiological, genomic and molecular mechanisms involved in boron tolerance in B. rapa. Initially, B. rapa genotypes were screened for tolerance to boron toxicity using hydroponic and soil assays. On the basis of primary root length, severity of leaf toxicity symptoms, dry matter accumulation and shoot boron uptake, the B. rapa genotypes WWY Sarson and Local were identified as the most tolerant and the B. rapa genotypes Shillong and Kaga the most susceptible to toxic boron concentrations (1000 ?M B in hydroponic assay; 54 mg B kg-1 soil in soil assay). The main mechanism of tolerance to boron toxicity in B. rapa involved reduced net boron uptake by roots, with some boron accumulation in the tap roots and partial exclusion of boron from shoots. Furthermore, boron uptake was much lower in the WWY Sarson and Local genotypes than in the Shillong genotype, despite higher rates of transpiration. This implied that an active boron efflux mechanism may be operating in the tolerant genotypes. The inheritance pattern of tolerance to boron toxicity in B. rapa genotype, WWY Sarson best fitted a Mendelian model of two major dominant and epistatic genes. A B. rapa linkage map was constructed from an intraspecific F2 population (WWY Sarson X Shillong) with ISSR, RAPD, SRAP and SSR marker loci. The linkage map spanned a total length of 874.1 cM and contained 12 linkage groups. Chisquare analysis (P < 0.05) revealed 25 dominant markers that showed segregation distortion in the F2 progeny. QTL analysis using composite interval analysis identified three significant peaks on LG2 and LG8 that were associated with primary root length and which accounted for 17% of the trait variation. Differential transcript analysis of SRAP markers following exposure to a toxic boron concentration identified up-regulation of me4+em2570bp, me2+em2650bp, me2+em1 1600bp, me2+em1800bp and me4+em2500bp genes in Shillong and Kaga and down-regulation of me2+em2650bp, me2+em1 1600bp, me2+em1800bp and me1+em21200bp genes in WWY Sarson and Local. Of these, a UDP-glycosyltransferase gene (sharing 80% similarity to the Arabidopsis thaliana homolog) was highly transcribed only in the sensitive genotype, Shillong, and may be involved in excessive boron cross-linking to the glycosyl groups present in the cell walls and/or membranes eventually causing the observed reductions in shoot and root growth