Analysis of QTL DM4.1 for Downy Mildew Resistance in Cucumber Reveals Multiple subQTL: A Novel RLK as Candidate Gene for the Most Important subQTL

Jeroen A Berg; Freddy W K Hermans; Frank Beenders; Lina Lou; Wim H Vriezen; Richard G F Visser; Yuling Bai; Henk J Schouten

doi:10.3389/fpls.2020.569876

Analysis of QTL DM4.1 for Downy Mildew Resistance in Cucumber Reveals Multiple subQTL: A Novel RLK as Candidate Gene for the Most Important subQTL

Front Plant Sci. 2020 Oct 22:11:569876. doi: 10.3389/fpls.2020.569876. eCollection 2020.

Authors

Jeroen A Berg¹, Freddy W K Hermans², Frank Beenders², Lina Lou^{1

3}, Wim H Vriezen², Richard G F Visser¹, Yuling Bai¹, Henk J Schouten¹

Affiliations

¹ Plant Breeding, Wageningen University & Research, Wageningen, Netherlands.
² Nunhems Netherlands BV, Nunhem, Netherlands.
³ Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, China.

Abstract

One of the biggest problems in cucumber cultivation is cucurbit downy mildew (DM), caused by the obligate biotroph Pseudoperonospora cubensis. Whereas DM in cucumber was previously efficiently controlled by the dm-1 gene from Indian cucumber accession PI 197087, this resistance was broken by new DM strains, prompting the search for novel sources of resistance. A promising source of resistance is the wild cucumber accession PI 197088. It was previously shown that DM resistance in this genotype inherits polygenically. In this paper, we put the focus on one of the QTL, DM4.1 that is located on chromosome 4. QTL DM4.1 was shown to consist of three subQTL: DM4.1.1 affected pathogen-induced necrosis, DM4.1.2 was shown to have an additive effect on sporulation, and DM4.1.3 had a recessive effect on chlorosis as well as an effect on sporulation. Near-isogenic lines (NILs) were produced by introgressing the subQTLs into a susceptible cucumber line (HS279) with good horticultural traits. Transcriptomic analysis revealed that many genes in general, and defense pathway genes in particular, were differentially expressed in NIL DM4.1.1/.2 compared to NIL DM4.1.3 and the susceptible parent HS279. This indicates that the resistance from subQTL DM4.1.1 and/or subQTL DM4.1.2 likely involves defense signaling pathways, whereas resistance due to subQTL DM4.1.3 is more likely to be independent of known defense pathways. Based on fine-mapping data, we identified the RLK gene CsLRK10L2 as a likely candidate for subQTL DM4.1.2, as this gene was found to have a loss-of-function mutation in the susceptible parent HS279, and was strongly upregulated by P. cubensis inoculation in NIL DM4.1.1/.2. Heterologous expression of this gene triggered necrosis, providing further evidence that this gene is indeed causal for subQTL DM4.1.2.

Keywords: PI 197088; QTL mapping; cucumber (Cucumis sativus); downy mildew (Pseudoperonospora cubensis); leaf rust kinase 10-like (LRK10L); plant–pathogen interactions; receptor-like kinase (RLK); transcriptomics.