Abstract
Resistance to multiple races of Fusarium wilt is a desired trait in lettuce (Lactuca sativa L.) cultivars, as it is directly related to grower profitability. We focused on its resistance to the race 2 pathogen, and analyzed F2 individuals obtained from a cross between ‘VI185’ (resistance to race 2) and ‘ShinanoGreen’ (susceptible to race 2) for genetic mapping of its resistance locus. According to the ddRAD-seq analysis and resistance genotype prediction in each F2 individual by infection assay of derived F3 generation, a single semi-dominant locus on LG1 (qFOL1.2) was found to determine race 2 resistance, and a marker designated as LG1_v8_117.181Mbp showed complete co-segregation with the resistance phenotype. Fine mapping by identified PCR-based markers in this locus allowed to show that qFOL1.2 were located in the position of 116.468–117.974Mbp in LG1. Association of the resistance phenotypes and genotypes of the PCR-based markers for qFOL1.2 in 42 lettuce cultivars showed that genotypes of all the analyzed markers were uniform in all the susceptible cultivars, while their genotypes in the resistant cultivars were not always consistent. Nonetheless, only the genotype of LG1_v8_116.506Mbp was identical across all the 25 resistant cultivars. Thus, we defined that LG1_v8_116.506Mbp was a broadly available marker for selection of race 2-resistance in lettuce cultivars. Our results provide additional breeding tool for resistance to race 2, and can accelerate pyramiding of resistance loci to multiple races of Fusarium wilt in lettuce breeding program.
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Data availability
All data analyzed during this study are included in this published article and its supplementary information files.
Code availability
Scripts for these analyses are deposited in https://github.com/KousukeSEKI/RAD-seq_scripts as (1) Parent_script.R, (2) RAD_mapping_script_BWA_aln_script.txt, (3) Map_list_script.R, (4) Population_tagcount_script.R, (5) Genotype_script.R, (6) Convert_ABH_script.R, and (7) Post_data_correction_script.R.
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Acknowledgements
This work was supported by Cooperative Research Grant of the Genome Research for BioResource, NODAI Genome Research Center, Tokyo University of Agriculture. We thank Rumi Ohtake of NODAI Genome Research Center, Tokyo University of Agriculture, Dr. Kazuhiro Yagasaki and Hiroaki Serizawa of the Nagano Vegetable Ornamental Crops Experiment Station, Dr. Yoshifumi Yamaguchi in University of Hokkaido and Dr. Satoshi Watanabe in University of Saga for helpful discussion and advice. We also thank Chizuru Kagami, Yoko Takahashi, Yoshie Nakayama, and especially Atsushi Tsukada and Hideaki Okazawa for technical assistance in the field experiment.
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This work was supported by Cooperative Research Grant of the Genome Research for BioResource, NODAI Genome Research Center, Tokyo University of Agriculture.
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KS, KK and HM planned the experiments. KS and MH performed the development of the mapping population. KS and MH performed bioassay for disease resistance. KS and YU performed the development of the ddRAD-seq library. KK and KT performed sequence analysis. KS and HM performed data analysis. KS wrote the manuscript. All authors read and approved the final manuscript.
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Seki, K., Komatsu, K., Hiraga, M. et al. Development of PCR-based marker for resistance to Fusarium wilt race 2 in lettuce (Lactuca sativa L.). Euphytica 217, 126 (2021). https://doi.org/10.1007/s10681-021-02857-1
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DOI: https://doi.org/10.1007/s10681-021-02857-1