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Korean Brassica oleracea germplasm offers a novel source of qualitative resistance to blackleg disease

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Abstract

Blackleg disease, caused by the hemibiotrophic fungal pathogen Leptosphaeria maculans, is one of the most devastating disease of Brassica species worldwide. To date, a total of 20 race-specific blackleg resistance (R) genes have been reported and all of those loci are located in either the A or B genomes of various Brassica species. The B. oleracea genome (CC) shares a high ancestral synteny with the A genome of B. rapa, suggesting the presence of qualitative (race specific) resistance to blackleg disease is also possible in B. oleracea germplasm. In the present study the C genome of Korean B. oleracea germplasm was screened for the presence of blackleg R genes. Thirty-two inbred cabbage lines with unknown resistance profiles, along with five control B. napus lines with well-characterised race-specific R genes, were assessed for cotyledon resistance against two L. maculans isolates with known and highly-contrasting avirulence gene (Avr) profiles. Two cabbage accessions were identified which produced a strong resistance when challenged with either isolate, demonstrating the presence of effective blackleg R genes in the cabbage C genome. Additionally, 16 microsatellite markers linked to seven different R genes of the B. napus A genome were converted into markers for their homologous regions on the B. oleracea C genome. These markers were used to screen all B. oleracea lines to assess if the novel C genome R genes were syntenous to known R gene-homologous regions of the A genome. The resistant cabbage lines offer C genome R genes for the protection of B. oleracea varieties against incursion of blackleg disease, as well as novel additional resistance sources for introgression into B. napus and B. carinata breeding material.

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Acknowledgments

We thank the Asia Seed Co., Ltd., Republic of Korea for providing B. oleracea seeds. This study was supported by the Golden Seed Project (Center for Horticultural Seed Development) of the Ministry of Agriculture, Food and Rural Affairs in the Republic of Korea (MAFRA).

Author contributions

ISN, JIP and NUA conceived the study. IP provided microsatellite marker information; HB and NJL provided fungal isolates. AHKR designed and conducted the experiments. RL assisted AHKR in plant culture and management, sample preparation, DNA isolation, purification, and cloning. AHKR analyzed the data and wrote the manuscript. NJL, HB and IP edited the manuscript. All authors read the article and approved the manuscript.

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Authors and Affiliations

  1. Department of Horticulture, Sunchon National University, Suncheon, South Korea

    Arif Hasan Khan Robin, Rawnak Laila, Jong-In Park, Nasar Uddin Ahmed & Ill-Sup Nou

  2. Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon, SK, Canada

    Nicholas J. Larkan, Hossein Borhan & Isobel A. P. Parkin

  3. Armatus Genetics Inc., Saskatoon, SK, Canada

    Nicholas J. Larkan

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  1. Arif Hasan Khan Robin
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Fig. S1

Percent infected lobe of the individual cotyledons of the seedlings of 32 cabbage lines and five B. napus control lines 10 days post-inoculation with isolates 03–02 s and 00–100 s of L. maculans, respectively. Lines 33–37 respectively represent B. napus susceptible line TopasDH16516 and four introgresion lines, Topas-Rlm1, Topas-Rlm2, Topas-Rlm3 and Topas-Rlm4. Lines with more than 60% lobe diameter infected were considered susceptible. (DOCX 105 kb)

Fig. S2

Progression of blackleg disease infection (a) caused by 03–02 s isolate and (b) 00–100 s isolate in R (in squared boxes) and S lines (L) of cabbage. (DOCX 1414 kb)

Fig. S3

Banding profile of 32 cabbage lines in agarose gel electrophoresis (white bands in black background) and in QIAxcel (black bands in white background). Differences in band sizes were recorded from QIAxcel. (DOCX 7268 kb)

Fig. S4

Cloned and sequenced microsatellite marker regions of cabbage lines. (DOCX 33 kb)

Table S1

Thirty two cabbage inbred lines and five B. napus control lines used to screen for existence of Leptosphaeria maculans resistance gene in Brassica oleracea L. (DOCX 21 kb)

Table S2

Selected Resistant (R) and susceptible (S) cabbage inbred lines used to follow disease reactions at the seedling stage against two Leptosphaeria maculans isolates. (DOCX 20 kb)

Table S3

SSR markers with primer sequence collected from literature targeted to potential blackleg resistance genes in Brassica oleracea. Primer sequences were compared with the assembled genome sequence of B. oleracea using BLAST (Altschul et al. 1990). (DOCX 24 kb)

Table S4

Interaction between bioassay results with L. maculans isolates (a) 03–02 s and (b) (a) 00–100 s and microsatellite marker bands against specific resistant genes for 32 cabbage lines. R, resistant; MR, moderately resistant; I, intermediate; S, susceptible and HS, highly susceptible indicate status of interactions between each cabbage line and each fungal isolate. Plus (+) sign favours resistant polymorphic marker type and minus (−) sign indicates absence of resistant marker type. (DOCX 31 kb)

Table S5

(DOCX 21 kb)

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Robin, A.H.K., Larkan, N.J., Laila, R. et al. Korean Brassica oleracea germplasm offers a novel source of qualitative resistance to blackleg disease. Eur J Plant Pathol 149, 611–623 (2017). https://doi.org/10.1007/s10658-017-1210-0

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