Genome-wide association study to identify candidate loci and genes for Mn toxicity tolerance in rice.

Manganese (Mn) is an essential micro-nutrient for plants, but flooded rice fields can accumulate high levels of Mn2+ leading to Mn toxicity. Here, we present a genome-wide association study (GWAS) to identify candidate loci conferring Mn toxicity tolerance in rice (Oryza sativa L.). A diversity panel of 288 genotypes was grown in hydroponic solutions in a greenhouse under optimal and toxic Mn concentrations. We applied a Mn toxicity treatment (5 ppm Mn2+, 3 weeks) at twelve days after transplanting. Mn toxicity caused moderate damage in rice in terms of biomass loss and symptom formation despite extremely high shoot Mn concentrations ranging from 2.4 to 17.4 mg g-1. The tropical japonica subpopulation was more sensitive to Mn toxicity than other subpopulations. Leaf damage symptoms were significantly correlated with Mn uptake into shoots. Association mapping was conducted for seven traits using 416741 single nucleotide polymorphism (SNP) markers using a mixed linear model, and detected six significant associations for the traits shoot manganese concentration and relative shoot length. Candidate regions contained genes coding for a heavy metal transporter, peroxidase precursor and Mn2+ ion binding proteins. The significant marker SNP-2.22465867 caused an amino acid change in a gene (LOC_Os02g37170) with unknown function. This study demonstrated significant natural variation in rice for Mn toxicity tolerance and the possibility of using GWAS to unravel genetic factors responsible for such complex traits.

Responses of contrasting rice genotypes to excess manganese and their implications for lignin synthesis.

Manganese (Mn) toxicity is frequently encountered in crops grown on soils with low pH or low redox potential, and harmful to plant development and growth. This study aimed at exploring adaptive mechanisms to Mn toxicity in rice, and investigated the effects of Mn toxicity on shoot lignification. Sixteen rice genotypes were grown in hydroponic solutions and exposed to normal (0.5 mg dm-3) or toxic (5 mg dm-3) Mn concentrations for three weeks. Morphological responses to Mn toxicity included a significant reduction in shoot length and the formation of visible symptoms scored as leaf damage index (LDI). Based on shoot Mn concentrations in the Mn toxic treatment, genotypes were classified as Mn includers and excluders. Across different genotypes, shoot Mn concentrations were significantly negatively correlated with relative shoot length and positively correlated with LDI. Consequently, the most tolerant genotypes in terms of morphology were all excluders, while the most sensitive genotypes were includers. The sensitive genotypes were also more responsive to manganese in terms of lipid peroxidation than tolerant genotypes. Shoots of rice plants grown in the high Mn treatment showed a higher level of lignification measured as thioglycolic acid lignin (TGAL), especially among Mn includers. TGAL was positively correlated with shoot Mn concentration and the levels of phenolics. In contrast, peroxidase activity was not responsive to the Mn treatment and was not significantly correlated with shoot lignification. In conclusion, exclusion is a dominant tolerance mechanism to Mn toxicity in rice. Further, Mn stimulated lignin biosynthesis in rice, especially in genotypes that were unable to exclude Mn.