Genetic research in a public-private research consortium: prospects for indirect use of Elige breeding germplasm in academic research

The creation of a public¿private research partnership between plant breeding industry and academia can be beneficial for all parties involved. Academic partners benefit from the material contributions by industry and a practically relevant research focus, while industry benefits from increased insights and methodology tailored to a relevant set of data. However, plant breeding industry is highly competitive and there are obvious limits to the data and material partners are willing and able to share. This will usually include current and historic released cultivated materials, but will very often not include the elite germplasm used in-house to create new cultivars. Especially for crops where hybrid cultivars dominate the market, parental lines of hybrid cultivars are considered core assets that are never provided to outside parties. However, this limitation often does not apply to DNA or genetic fingerprints of these parental lines. We developed a procedure to take advantage of elite breeding materials for the creation of new promising research populations, through indirect selection of parents. The procedure starts with the identification of a number of traits for further study based on the presence of marker-trait associations and a priori knowledge within the participating companies about promising traits for quality improvement. Next, regression-based multi-QTL models are fitted to hybrid cultivar data to identify QTLs. Fingerprint data of parental lines of a limited number of specific hybrids are then used to predict parental phenotypes using the multi-QTL model fitted on hybrid data. The specific hybrids spanned the whole of the sensory space adequately. Finally, a choice of parental lines is made based on the QTL model predictions and new promising line combinations are identified. Breeding industry is then asked to create and provide progeny of these line combinations for further research. This approach will be illustrated with a case study in tomato

Normal adult survival but reduced Bemisia tabaci oviposition rate on tomato lines carrying an introgression from S. habrochaites

Background Host plant resistance has been proposed as one of the most promising approaches in whitefly management. Already in 1995 two quantitative trait loci (Tv-1 and Tv-2) originating from S. habrochaites CGN1.1561 were identified that reduced the oviposition rate of the greenhouse whitefly (Trialeurodes vaporariorum). After this first study, several others identified QTLs affecting whitefly biology as well. Generally, the QTLs affecting oviposition were highly correlated with a reduction in whitefly survival and the presence of high densities of glandular trichomes type IV. The aim of our study was to further characterize Tv-1 and Tv-2, and to determine their role in resistance against Bemisia tabaci. Results We selected F2 plants homozygous for the Tv-1 and Tv-2 QTL regions and did three successive backcrosses without phenotypic selection. Twenty-three F2BC3 plants were phenotyped for whitefly resistance and differences were found in oviposition rate of B. tabaci. The F2BC3 plants with the lowest oviposition rate had an introgression on Chromosome 5 in common. Further F2BC4, F2BC4S1 and F2BC4S2 families were developed, genotyped and phenotyped for adult survival, oviposition rate and trichome type and density. It was possible to confirm that an introgression on top of Chr. 5 (OR-5), between the markers rs-2009 and rs-7551, was responsible for reducing whitefly oviposition rate. Conclusion We found a region of 3.06 Mbp at the top of Chr. 5 (OR-5) associated with a reduction in the oviposition rate of B. tabaci. This reduction was independent of the presence of the QTLs Tv- 1 and Tv-2 as well as of the presence of trichomes type IV. The OR-5 locus will provide new opportunities for resistance breeding against whiteflies, which is especially relevant in greenhouse cultivation

Normal adult survival but reduced Bemisia tabaci oviposition rate on tomato lines carrying an introgression from S. habrochaites

Background Host plant resistance has been proposed as one of the most promising approaches in whitefly management. Already in 1995 two quantitative trait loci (Tv-1 and Tv-2) originating from S. habrochaites CGN1.1561 were identified that reduced the oviposition rate of the greenhouse whitefly (Trialeurodes vaporariorum). After this first study, several others identified QTLs affecting whitefly biology as well. Generally, the QTLs affecting oviposition were highly correlated with a reduction in whitefly survival and the presence of high densities of glandular trichomes type IV. The aim of our study was to further characterize Tv-1 and Tv-2, and to determine their role in resistance against Bemisia tabaci. Results We selected F2 plants homozygous for the Tv-1 and Tv-2 QTL regions and did three successive backcrosses without phenotypic selection. Twenty-three F2BC3 plants were phenotyped for whitefly resistance and differences were found in oviposition rate of B. tabaci. The F2BC3 plants with the lowest oviposition rate had an introgression on Chromosome 5 in common. Further F2BC4, F2BC4S1 and F2BC4S2 families were developed, genotyped and phenotyped for adult survival, oviposition rate and trichome type and density. It was possible to confirm that an introgression on top of Chr. 5 (OR-5), between the markers rs-2009 and rs-7551, was responsible for reducing whitefly oviposition rate. Conclusion We found a region of 3.06 Mbp at the top of Chr. 5 (OR-5) associated with a reduction in the oviposition rate of B. tabaci. This reduction was independent of the presence of the QTLs Tv- 1 and Tv-2 as well as of the presence of trichomes type IV. The OR-5 locus will provide new opportunities for resistance breeding against whiteflies, which is especially relevant in greenhouse cultivation

QRLs for tomato powdery mildew resisance (Oidium lycopersici) in Lycopersicon parviflorum G1.1601 colocalize with two qualitative powdery mildew resistance genes

Tomato (Lycopersicon esculentum) is susceptible to the powdery mildew Oidium lycopersici, but several wild relatives such as Lycopersicon parviflorum G1.1601 are completely resistant. An F-2 population from a cross of Lycopersicon esculentum cv. Moneymaker x Lycopersicon parviflorum G1.1601 was used to map the O. lycopersici resistance by using amplified fragment length polymorphism markers. The resistance was controlled by three quantitative trait loci (QTLs). Ol-qtl1 is on chromosome 6 in the same region as the Ol-1 locus, which is involved in a hypersensitive resistance response to O. lycopersici. Ol-qtl2 and Ol-qtl3 are located on chromosome 12, separated by 25 cM, in the vicinity of the Lv locus conferring resistance to another powdery mildew species, Leveillula taurica. The three QTLs, jointly explaining 68% of the phenotypic variation, were confirmed by testing F-3 progenies. A set of polymerase chain reaction-based cleaved amplified pollymorphic sequence and sequence characterized amplified region markers was generated for efficient monitoring of the target QTL genomic regions in marker assisted selection. The possible relationship between genes underlying major and partial resistance for tomato powdery mildew is discussed

Tomato defense to Oidium neolycopersici: Dominant Ol genes confer isolate-dependent resistance via a different mechanism than recessive ol-2

Tomato powdery mildew caused by Oidium neolycopersici has become a globally important disease of tomato (Lycopersicon esculentum). To study the defense responses of tomato triggered by tomato powdery mildew, we first mapped a set of resistance genes to O. neolycopersici from related Lycopersicon species. An integrated genetic map was generated showing that all the dominant resistance genes (Ol-1, Ol-3, Ol-4, Ol-5, and Ol-6) are located on tomato chromosome 6 and are organized in three genetic loci. Then, near-isogenic lines (NIL) were produced that contain the different dominant Ol genes in a L. esculentum genetic background. These NIL were used in disease tests with local isolates of O. neolycopersici in different geographic locations, demonstrating that the resistance conferred by different Ol genes was isolate-dependent and, hence, may be race-specific. In addition, the resistance mechanism was analyzed histologically. The mechanism of resistance conferred by the dominant Ol genes was associated with hypersensitive response, which varies in details depending on the Ol-gene in the NIL, while the mechanism of resistance governed by the recessive gene ol-2 on tomato chromosome 4 was associated with papillae formatio

Exploration of a Resequenced Tomato Core Collection for Phenotypic and Genotypic Variation in Plant Growth and Fruit Quality Traits

A tomato core collection consisting of 122 gene bank accessions, including landraces, old cultivars, and wild relatives, was explored for variation in several plant growth, yield and fruit quality traits. The resequenced accessions were also genotyped with respect to a number of mutations or variations in key genes known to underlie these traits. The yield-related traits fruit number and fruit weight were much higher in cultivated varieties when compared to wild accessions, while, in wild tomato accessions, Brix was higher than in cultivated varieties. Known mutations in fruit size and shape genes could well explain the fruit size variation, and fruit colour variation could be well explained by known mutations in key genes of the carotenoid and flavonoid pathway. The presence and phenotype of several plant architecture affecting mutations, such as self-pruning (sp), compound inflorescence (s), jointless-2 (j-2), and potato leaf (c) were also confirmed. This study provides valuable phenotypic information on important plant growth- and quality-related traits in this collection. The allelic distribution of known genes that underlie these traits provides insight into the role and importance of these genes in tomato domestication and breeding. This resource can be used to support (precision) breeding strategies for tomato crop improvement

Phenotyping of a diverse tomato collection for postharvest shelf-life.

In all fruit and vegetable crops, reduction in quality during postharvest storage leads to substantial losses of primary production with enormous economic consequences. Also in tomato, fruit shelf-life is an important quality trait. In this study a collection of tomato accessions, consisting of 92 S. lycopersicum landraces and old cultivars and several S. pimpinellifolium accessions, was phenotyped for several shelf-life parameters and biochemical characterization was performed during the postharvest shelf-life of fruit from selected accessions. This collection was selected based on available genotypic data and represents the genetic diversity present in the EU–SOL tomato core collection (Roohanitaziani, 2020). The core collection was grown in a greenhouse, and fruit were harvested at the breaker-turning stage and stored in a controlled climate chamber for 42 d at 18 ⁰C. The shelf-life attributes firmness loss, weight loss, as well as color pigments, were measured once a week and evaluated over time. All three shelf-life-related parameters varied markedly among accessions, resulting in fruit with different shelf-life. The most promising accessions of the first screen were re-grown and analyzed to validate the initial results and six accessions with contrasting shelf-life were selected for metabolite analysis. Fruit were harvested at the breaker stage and stored for 35 d at 18 ⁰C. Samples were taken at weekly intervals and analyzed for volatile compounds, primary metabolites and cell wall polysaccharide monomers. During storage long and short shelf-life accessions showed considerable differences in their content of sugars, such as galactose and polyamines, such as putrescine in their pericarp. The content of three cell wall sugars, galactose, arabinose and galacturonic acid, underwent considerable changes during postharvest storage. The short shelf-life accessions contained a higher amount of arabinose and galactose in their cell wall than other accessions which is indicative of highly branched pectin. This knowledge provides a better understanding of the difference in pectin structure between short and long shelf-life fruit during the ripening process

Quantitative resistance to Botrytis cinerea from Solanum neorickii

Tomato (Solanum lycopersicum) is susceptible to gray mold (Botrytis cinerea). Quantitative resistance to B. cinerea was previously identified in a wild relative, S. neorickii G1.1601. The 122 F3 families derived from a cross between the susceptible S. lycopersicum cv. Moneymaker and the partially resistant S. neorickii G1.1601 were tested for susceptibility to B. cinerea using a stem bioassay. Three putative quantitative trait loci (pQTL) were detected: pQTL3 and pQTL9 reducing lesion growth (LG) and pQTL4 reducing disease incidence (DI). For each pQTL, a putative homologous locus was identified recently in another wild tomato relative, S. habrochaites LYC4. pQTL3 was confirmed by assessing disease resistance in BC3S1 and BC3S2 progenies of S. neorickii G1.1601. pQTL4 was not statistically confirmed but the presence of the S. neorickii resistance allele reduced DI in all three tested populations. The reduction in LG of pQTL9 was not confirmed but rather, this locus conferred a reduced DI, similar to observations in the QTL study using S. habrochaites. The results are discussed in relation to other disease resistance loci identified in studies with other wild tomato relatives