ELR is a true pattern recognition receptor that associates with elicitins from diverse Phytophthora species

The first layer of plant immunity against pathogens is mediated by cell surface pattern recognition receptors (PRRs) that recognize pathogen molecules in the apoplast. Several pairs of PRRs and their matching extracellular ligands have been described but, in many cases, actual evidence for ligand binding by the PRR is lacking. The receptor-like protein ELR from Solanum microdontum, which triggers cell death upon co-expression with elicitins of various Phytophthora species and enhances resistance to late blight caused by Phytophthora infestans, was previously identified as the elicitin receptor by forward genetic screenings employing the INF1 elicitin of P. infestans. In this study, we investigated whether ELR associates with INF1 and other elicitins that are secreted by diverse Phytophthora spp. We performed in planta and in vitro co-immunoprecipitation of ELR with several affinity-tagged elicitins, as well as in planta transient co-expression assays. We found that ELR physically interacts with the class I elicitins INF1 and ParA1, from P. infestans and Phytophthora parasitica, respectively, which is in line with their ability to cause cell death when co-expressed with ELR in potato. Together, we demonstrate that ELR is a genuine PRR that binds elicitins of Phytophthora species.

Detection of Novel QTLs for Late Blight Resistance Derived from the Wild Potato Species Solanum microdontum and Solanum pampasense

Wild potato species continue to be a rich source of genes for resistance to late blight in potato breeding. Whilst many dominant resistance genes from such sources have been characterised and used in breeding, quantitative resistance also offers potential for breeding when the loci underlying the resistance can be identified and tagged using molecular markers. In this study, F1 populations were created from crosses between blight susceptible parents and lines exhibiting strong partial resistance to late blight derived from the South American wild species Solanum microdontum and Solanum pampasense. Both populations exhibited continuous variation for resistance to late blight over multiple field-testing seasons. High density genetic maps were created using single nucleotide polymorphism (SNP) markers, enabling mapping of quantitative trait loci (QTLs) for late blight resistance that were consistently expressed over multiple years in both populations. In the population created with the S. microdontum source, QTLs for resistance consistently expressed over three years and explaining a large portion (21–47%) of the phenotypic variation were found on chromosomes 5 and 6, and a further resistance QTL on chromosome 10, apparently related to foliar development, was discovered in 2016 only. In the population created with the S. pampasense source, QTLs for resistance were found in over two years on chromosomes 11 and 12. For all loci detected consistently across years, the QTLs span known R gene clusters and so they likely represent novel late blight resistance genes. Simple genetic models following the effect of the presence or absence of SNPs associated with consistently effective loci in both populations demonstrated that marker assisted selection (MAS) strategies to introgress and pyramid these loci have potential in resistance breeding strategies.

RLP/K enrichment sequencing; a novel method to identify receptor-like protein (RLP) and receptor-like kinase (RLK) genes

SummaryThe identification of immune receptors in crop plants is time-consuming but important for disease control. Previously, resistance gene enrichment sequencing (RenSeq) was developed to accelerate mapping of nucleotide-binding domain and leucine-rich repeat containing (NLR) genes. However, resistances mediated by pattern recognition receptors (PRRs) remain less utilised.Here, our pipeline shows accelerated mapping of PRRs. Effectoromics leads to precise identification of plants with target PRRs, and subsequent RLP/K enrichment sequencing (RLP/KSeq) leads to detection of informative SNPs that are linked to the trait.Using Phytophthora infestans as a model, we identified Solanum microdontum plants that recognize the apoplastic effectors INF1 or SCR74. RLP/KSeq in a segregating Solanum population confirmed the localization of the INF1 receptor on chromosome 12, and lead to the rapid mapping of the response to SCR74 to chromosome 9. By using markers obtained from RLP/KSeq in conjunction with additional markers, we fine-mapped the SCR74 receptor to a 43-kbp G-LecRK locus.Our findings show that RLP/KSeq enables rapid mapping of PRRs and is especially beneficial for crop plants with large and complex genomes. This work will enable the elucidation and characterisation of the non-NLR plant immune receptors and ultimately facilitate informed resistance breeding.

Micronutrient concentration in potato clones with distinct physiological sensitivity to Al stress

The objective of this study was to evaluate the effects of aluminum (Al) on the zinc (Zn), manganese (Mn), iron (Fe) and copper (Cu) concentrations in four potato clones (Macaca and Dakota Rose: both Al-sensitive clones; and SMIC148-A and Solanum microdontum: both Al-tolerant-clones), grown in a nutrient solution (pH 4.00) with 0, 50, 100, 150 and 200mg Al L-1. Root Zn and Fe concentrations decreased linearly with the increase of Al levels in Macaca, SMIC148-A and Dakota Rose and increased linearly in S. microdontum. Shoot Zn concentration showed a quadratic relationship with Al in S. microdontum and SMIC148-A, but a curvilinear response in Dakota Rose. Shoot Fe concentration showed a quadratic relationship with Al in S. microdontum, SMIC148-A and Dakota Rose. Root Mn concentration decreased linearly in Macaca and SMIC148-A, and increased linearly in S. microdontum with Al levels. Mn concentration showed a quadratic relationship with Al in roots of Dakota Rose and in shoot of SMIC148-A, and increased curvilinearly with Al levels in shoot of Dakota Rose. In shoot, there was no alteration in Zn, Fe and Mn in Macaca and Mn concentration in S. microdontum. Roots and shoot Cu concentration increased linearly in Dakota Rose, and showed quadratic relationship with Al in Macaca. Roots Cu concentration showed a quadratic relationship with Al levels in S. microdontum and SMIC148-A. Shoot Cu concentration increased linearly in S. microdontum, and decreased linearly in SMIC148-A. Therefore, the excessive Al accumulation affected the uptake and distribution of Zn, Fe, Mn and Cu in roots and shoot of potato clones.The response of shoot Cu concentration to Al was less altered in the Al-tolerant clones than was in Al-sensitive clones. Aluminum tolerance in S. microdontum may be connected with greater levels of Zn, Fe and Mn in the roots.

The RPi-mcd1 Locus from Solanum microdontum Involved in Resistance to Phytophthora infestans, Causing a Delay in Infection, Maps on Potato Chromosome 4 in a Cluster of NBS-LRR Genes

The distinction between field resistance and resistance based on resistance (R) genes has been proven valid for many plant–pathogen interactions. This distinction does not seem to be valid for the interaction between potato and late blight. In this study, a locus involved in late blight resistance, derived from Solanum microdontum, provides additional evidence for this lack of distinction. The resistance is associated with a hypersensitive response and results in a delay of infection of approximately 1 to 2 weeks. Both a quantitative as well as a qualitative genetic approach were used, based on data from a field assay. Quantitative trait locus (QTL) analysis identified a QTL on chromosome 4 after correction of the resistance data for plant maturity. A qualitative genetic analysis resulted in the positioning of this locus on the short arm of chromosome 4 in between amplified fragment length polymorphism marker pCTmACG_310 and cleaved amplified polymorphic sequence markers TG339 and T0703. This position coincides with a conserved Phytophthora R gene cluster which includes R2, R2-like, RPi-blb3, and RPi-abpt. This implies that RPi-mcd1 is the fifth R gene of this nucleotide-binding site leucine-rich repeat cluster. The implications of our results on R-gene-based and field resistance are discussed.

Physiological and oxidative stress responses of four potato clones to aluminum in nutrient solution

Aluminum toxicity is a serious problem in Brazilian soils and selecting potato clones is an important strategy to produce this crop on these kinds of soils. Potato clones, Macaca, SMIC148-A, Dakota Rose, and Solanum microdontum, were grown in a nutrient solution (pH 4.0) with 0, 50, 100, 150 and 200 mg Al L-1. After 7 d, Al concentration in both root system and shoot of all clones increased linearly with increasing Al levels. Based on relative root growth, S. microdontum and SMIC148-A were considered Al-tolerant clones, whereas Macaca and Dakota Rose were considered Al-sensitive. Shoot growth in Macaca linearly decreased with increasing Al levels. Root H2O2 concentration in both Al-sensitive clones increased with increasing Al supply, whereas in Al-tolerant clones it either decreased (SMIC148-A) or demonstrated no alteration (S. microdontum). Shoot H2O2 concentration increased linearly in Macaca, whereas for Dakota Rose it showed a quadratic relationship with Al levels. On the other hand, shoot H2O2 concentration in the Al-tolerant clones either demonstrated no alteration (S. microdontum) or presented lower levels (SMIC148-A). Root catalase (CAT) activity in both Al-sensitive clones increased with increasing Al levels, whereas in Al-tolerant clones it either demonstrated no alteration (SMIC148-A) or presented lower levels (S. microdontum). Shoot CAT activity in the S. microdontum increased curvilinearly with increasing Al levels. In all potato clones, chlorophyll concentration showed a curvilinear response to Al supply, where in Al-sensitive clones it decreased upon addition of Al exceeding 100 mg L-1, but in SMIC148-A it increased at levels between approximately 100 and 150 mg L-1, and decreased in S. microdontum regardless of the Al level. Carotenoid concentrations in the Al-sensitive clones were linearly decreased with increasing Al levels. Aluminum supply caused root lipid peroxidation only in the Al-sensitive clones, whereas in the shoot it increased linearly in the Al-sensitive clones and in S. microdontum it only increased at around 50 mg L-1. Most of root protein oxidation was only observed in the Al-sensitive clones. However, shoot protein oxidation was increased with increasing Al levels for all potato clones. These results indicate that oxidative stress caused by Al in potato may harm several components of the cell, mainly in Al-sensitive clones.