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Using chlorate as an analogue to nitrate to identify candidate genes for nitrogen use efficiency in barley

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Abstract

Nitrogen (N) is one of the most important macronutrients for crop growth and development. Large amounts of N fertilizers are applied exogenously to improve grain yield and quality, which has led to environmental pollution and high cost of production. Therefore, improvement of N use efficiency (NUE) is a very important aspect for sustainable agriculture. Here, a pilot experiment was firstly conducted with a set of barley genotypes with confirmed NUE to validate the fast NUE screening, using chlorate as an analogue to nitrate. High NUE genotypes were susceptible to chlorate-induced toxicity whereas the low NUE genotypes were tolerant. A total of 180 barley RILs derived from four parents (Compass, GrangeR, Lockyer and La Trobe) were further screened for NUE. Leaf chlorosis induced by chlorate toxicity was the key parameter observed which was later related to low-N tolerance of the RILs. There was a distinct variation in chlorate susceptibility of the RILs with leaf chlorosis in the oldest leaf ranging from 10 to 80%. A genome-wide association study (GWAS) identified 9 significant marker-trait associations (MTAs) conferring high chlorate sensitivity on chromosomes 2H (2), 3H (1), 4H (4), 5H (1) and Un (1). Genes flanking with these markers were retrieved as potential targets for genetic improvement of NUE. Genes encoding Ferredoxin 3, leucine-rich receptor-like protein kinase family protein and receptor kinase are responsive to N stress. MTA4H5468 which exhibits concordance with high NUE phenotype can further be explored under different genetic backgrounds and successfully applied in marker-assisted selection.

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Acknowledgements

We acknowledge the Western Australian State Agricultural Biotechnology Centre (SABC), Murdoch University and Western Crop Genetics Alliance (WCGA) for providing us with facilities to conduct the experiments. We also thank Ms. Lee-Anne McFawn from the Department of Primary Industries and Regional Development (DPIRD), Western Australia, for her support in sorting out the seeds.

Funding

This project was made possible by the generous support of Murdoch University Western Australia. Sakura Karunarathne was awarded a Murdoch International Postgraduate Scholarship (MIPS).

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SK and TA conducted the phenotyping experiments. SK and VD conducted data analysis and interpretation. VD and X-QZ generated the genotype data. CL, YH and X-QZ conceived the project. SK drafted the manuscript with input from all authors.

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Correspondence to Chengdao Li.

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Karunarathne, S.D., Han, Y., Zhang, XQ. et al. Using chlorate as an analogue to nitrate to identify candidate genes for nitrogen use efficiency in barley. Mol Breeding 41, 47 (2021). https://doi.org/10.1007/s11032-021-01239-8

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  • DOI: https://doi.org/10.1007/s11032-021-01239-8

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