Diploid Potato Research Articles

Erratum to: Genotype–environment interaction and yield stability analysis of Colombian yellow diploid potato cultivars using mixed linear models and environmental covariates

Erratum to: Genotype–environment interaction and yield stability analysis of Colombian yellow diploid potato cultivars using mixed linear models and environmental covariates

Characterizing Diploid and Tetraploid Potato Cultivars with Reflectance Spectroscopy

Characterizing potato cultivars is essential for diagnosing the specific production and management requirements. Employing techniques that facilitate rapid, objective, and cost-effective identification of various aspects of plant physiology is crucial for generating this valuable information. In this study, our goal was to identify different potato cultivars across three distinct phenological phases utilizing reflectance spectroscopy within the visible and near-infrared ranges. Reflectance spectra were measured using a portable spectrometer on leaves obtained from five different potato cultivars. We constructed classification models to determine the accurate classification percentages for each cultivar within specific phenological stages. General trends in the reflectance spectra were observed, which were consistent across all five cultivars within the measured range. During the vegetative phase, there was a uniform reflectance of around 5% in the green region. This value experienced a slight decrease during the flowering phase and subsequently rose to approximately 12% during the senescence phase. In the near-infrared (NIR) range, a reduction in reflectance from 50 to 30% was noted during the flowering phase, followed by a cultivar-dependent increase towards senescence. The specific wavelengths identified in the spectra enabled the accurate classification of plants from each cultivar across the vegetative, flowering, and senescence phases. The classification rates were notably high, with a success rate of 87% for Var. Paola, 96% for Var. Paysandú, 93% for Var. Violeta, and 100% for cv. 448 and 440. The Var. Paola exhibited superior correct classification rates, ranging from 85% for Paysandú and Violeta, 90% for cv. 440, to 100% for cv. 448. The characterization of potato cultivars using spectroscopy techniques yields valuable information that serves as fundamental input for precision agriculture systems.

Genotype–environment interaction and yield stability analysis of Colombian yellow diploid potato cultivars using mixed linear models and environmental covariates

AbstractSeveral statistical methods have been developed to decompose the phenotypic response based mainly on the general means, the effect of the genotype, and its interaction with the environment, treating the environmental variance as a confounding factor. However, the estimation of the environmental effect from the traits evaluated would establish a dependency on the genotype, resulting in a lower fit and predictive potential of the model. The integration of environmental covariates into the models that associate edaphoclimatic characteristics with the traits of interest has been proposed, with the purpose of increasing the predictive potential and the variance contained by the models. The aim of this study was to evaluate the sensitivity of the traits of interest in yellow diploid potato (Solanum tuberosum L. Group: Phureja) crops to environmental covariates, selecting the most relevant covariates as parameters in empirical multiple regression models based on environmental variance. The results showed high yield variability due to covariates of the water component, while the quality traits were mainly affected by traits of the energetic and physicochemical components of the soil. The adjusted models explained the variance intrinsically due to the environment, reaching adjustments greater than 20%. Therefore, it is concluded that the traits have a high phenotypic sensitivity, and the incorporation of environmental covariates into the genotype‐by‐environment interaction analysis models could improve the understanding of the stability and adaptability of cultivars from the data obtained in multi‐environment trials.

Characterization of traditional small potato (Solanum tuberosum L.) cultivars for nutritional, quality traits and ploidy level

Traditional small potato cultivars of North Eastern region of India are an important food source, but no documentation is availableregarding their ploidy level and nutritional worth. The study was undertaken to explore these cultivars to determine their ploidy levelby flow cytometry method. Fluorescence-activated cell sorter (FACS) based method was used for the analysis of ploidy level and theknown diploid potato clone Solanum chacoense was used as the reference clone. The ploidy level of these cultivars was determined bycomparing the value of mean peak position of G1 of reference diploid clone with mean peak position of G1 of small potato cultivars.Results indicated that all the 11 small potato cultivars collected from the Northeastern region were tetraploid with an average genomesize of 837.38 Mb although they were suspected to be diploid based on the tuber characters and overall plant morphology. Thesecultivars were characterized by having anthocyanin pigmentation of newly emerged sprouts and eye on tuber, small and narrow leaf,long stolon length (average 7.38 cm) and high number (average 33.90/plant) of very small (average 7.41 g) tubers compared to thecommercial potato varieties. However, these cultivars surpassed India’s commercial tetraploid potato varieties for dry matter, starch,ascorbic acid, β-carotene and total phenol contents. Appreciably rich nutritional profile coupled with matching levels of ploidy not onlyenhanced the biodiversity of cultivated potato but also suggested the possibility of utilizing these cultivars in the breeding programmeto enhance nutrient and antioxidant contents.

Identification and mapping of Rpi-blb4 in diploid wild potato species Solanum bulbocastanum

More than 170 years after causing the potato famine in Ireland, late blight is still considered one of the most devastating crop diseases. Commercial potato breeding efforts depend on natural sources of resistance to protect crops from the rapidly evolving late blight pathogen, Phytophthora infestans. We have identified and mapped a novel broad-spectrum disease resistance gene effective against P. infestans from the wild, diploid potato species Solanum bulbocastanum. Diagnostic resistance gene enrichment sequencing (dRenSeq) was used to confirm the uniqueness of the identified resistance. RenSeq and GenSeq-based mapping of the resistance, referred to as Rpi-blb4, alongside recombinant screening, positioned the locus responsible for the resistance to potato chromosome 5. The interval spans approximately 2.3 Mb and corresponds to the DM reference genome positions of 11.25 and 13.56 Mb.

Dissected Leaf 1 encodes an MYB transcription factor that controls leaf morphology in potato.

The transcription factor StDL1 regulates dissected leaf formation in potato and the genotype frequency of recessive Stdl1/Stdl1, which results in non-dissected leaves, has increased in cultivated potatoes. Leaf morphology is a key trait of plants, influencing plant architecture, photosynthetic efficiency and yield. Potato (Solanum tuberosum L.), the third most important food crop worldwide, has a diverse leaf morphology. However, despite the recent identification of several genes regulating leaf formation in other plants, few genes involved in potato leaf development have been reported. In this study, we identified an R2R3 MYB transcription factor, Dissected Leaf 1 (StDL1), regulating dissected leaf formation in potato. A naturally occurring allele of this gene, Stdl1, confers non-dissected leaves in young seedlings. Knockout of StDL1 in a diploid potato changes the leaf morphology from dissected to non-dissected. Experiments in N. benthamiana and yeast show that StDL1 is a transcriptional activator. Notably, by calculating the genotype frequency of the Stdl1/Stdl1 in 373-potato accessions, we found that it increases significantly in cultivated potatoes. This work reveals the genetic basis of dissected leaf formation in potato and provides insights into plant leaf morphology.

Efficient Genomic Prediction of Yield and Dry Matter in Hybrid Potato.

There is an ongoing endeavor within the potato breeding sector to rapidly adapt potato from a clonal polyploid crop to a diploid hybrid potato crop. While hybrid breeding allows for the efficient generation and selection of parental lines, it also increases breeding program complexity and results in longer breeding cycles. Over the past two decades, genomic prediction has revolutionized hybrid crop breeding through shorter breeding cycles, lower phenotyping costs, and better population improvement, resulting in increased genetic gains for genetically complex traits. In order to accelerate the genetic gains in hybrid potato, the proper implementation of genomic prediction is a crucial milestone in the rapid improvement of this crop. The authors of this paper set out to test genomic prediction in hybrid potato using current genotyped material with two alternative models: one model that predicts the general combining ability effects (GCA) and another which predicts both the general and specific combining ability effects (GCA+SCA). Using a training set comprising 769 hybrids and 456 genotyped parental lines, we found that reasonable a prediction accuracy could be achieved for most phenotypes with both zero common parents (ρ=0.36-0.61) and one (ρ=0.50-0.68) common parent between the training and test sets. There was no benefit with the inclusion of non-additive genetic effects in the GCA+SCA model despite SCA variance contributing between 9% and 19% of the total genetic variance. Genotype-by-environment interactions, while present, did not appear to affect the prediction accuracy, though prediction errors did vary across the trial's targets. These results suggest that genomically estimated breeding values on parental lines are sufficient for hybrid yield prediction.

Homoeologous Chromosome Pairing and Alien Introgression in Backcrossing Progenies Derived from Hybrids Solanum tuberosum (+) Mexican 2x (1 EBN) B-Genome Potato Species

We applied a genomic in situ hybridization (GISH) to analyze the genomic constitution of and meiotic pairing in interspecific somatic hybrids, and in a wide subset of backcrossing derivatives (BC1–BC5), from three interspecific combinations involving the cultivated potato, Solanum tuberosum (AAAA genome), and three diploid (1 EBN) wild Mexican potato species (genome BB)—S. tarnii, S. pinnatisectum, and S. bulbocastanum. The theoretically expected genomic composition was detected in the somatic hybrids (AAAABB) and in the BC1 progeny (AAAAB), whereas in the subsequent BC2–BC4 generations, the partial loss of alien chromosomes was observed and almost all the BC5 genotypes showed a complete chromosome elimination of wild species. GISH revealed a homoeologous pairing between the chromosomes of the A- and the B-genomes in each of the hybrid progenies. Using GISH, we selected introgression lines with a single chromosome of the wild species in a potato genome background, as well as introgression lines with intergenomic recombinant chromosomes. Moreover, via molecular screening, BC hybrids with diagnostic markers for the R-genes conferring resistance to late blight disease and to the quarantine pest of the potato–Columbia root-knot nematode—were selected. The potential application of the results obtained for the planning of introgressive schemes directed to the breeding of advanced lines with multiple disease and pest resistance is discussed.

HT-B and S-RNase CRISPR-Cas9 double knockouts show enhanced self-fertility in diploid Solanum tuberosum.

The Gametophytic Self-Incompatibility (GSI) system in diploid potato (Solanum tuberosum L.) poses a substantial barrier in diploid potato breeding by hindering the generation of inbred lines. One solution is gene editing to generate self-compatible diploid potatoes which will allow for the generation of elite inbred lines with fixed favorable alleles and heterotic potential. The S-RNase and HT genes have been shown previously to contribute to GSI in the Solanaceae family and self-compatible S. tuberosum lines have been generated by knocking out S-RNase gene with CRISPR-Cas9 gene editing. This study employed CRISPR-Cas9 to knockout HT-B either individually or in concert with S-RNase in the diploid self-incompatible S. tuberosum clone DRH-195. Using mature seed formation from self-pollinated fruit as the defining characteristic of self-compatibility, HT-B-only knockouts produced little or no seed. In contrast, double knockout lines of HT-B and S-RNase displayed levels of seed production that were up to three times higher than observed in the S-RNase-only knockout, indicating a synergistic effect between HT-B and S-RNase in self-compatibility in diploid potato. This contrasts with compatible cross-pollinations, where S-RNase and HT-B did not have a significant effect on seed set. Contradictory to the traditional GSI model, self-incompatible lines displayed pollen tube growth reaching the ovary, yet ovules failed to develop into seeds indicating a potential late-acting self-incompatibility in DRH-195. Germplasm generated from this study will serve as a valuable resource for diploid potato breeding.

High-Density Linkage Map Constructed from a Skim Sequenced Diploid Potato Population Reveals Transmission Distortion and QTLs for Tuber Yield and Pollen Shed

The reinvention of potato, from a tetraploid clonal crop into a diploid seed-based hybrid crop, requires insight in the mutational load, recombination landscape, and the genetic basis of fertility. Genomics-based breeding and QTL discovery rely on efficient genotyping strategies such as skim sequencing, to gather genotypic information. The application of skim sequencing to full-sib population of non-inbred parents remains challenging. Here, we report on an R implementation of the OutcrossSeq pipeline for diploids. We applied this pipeline to a large diploid skim sequenced potato population. We used the resulting bin-markers for the construction of high-density parent specific linkage maps, highlighting variation in parental recombination rate and structural variations. We subsequently explored transmission ratio distortion and non-independent assortment of alleles, indicative of large-effect deleterious mutations. Finally, we identified QTLs for seedling tuber yield in pots and pollen shed. This study showcases the range of genetic analyses, from marker inference, identification of transmission ratio distortion, and linkage map construction to QTL mapping, resulting in new insights that contribute to breeding diploid potato.

Functional growth analysis of diploid potato varieties (Solanum tuberosum Phureja group)

The growth analysis is relevant to understanding and comparing in terms of physiological and productive behavior varieties. The study aims to determine the accumulation and distribution of dry matter through different phenological stages of development in four diploid potato varieties. For this, in a randomized complete block design with three replications, four varieties of diploid potato were evaluated: Paola, Violeta, Primavera, and Paysandú. Based on direct measurements of leaf area and dry matter of each organ, it was computed the growth indices of relative growth rate (RGR), net assimilation rate (NAR), leaf area relation (LAR), and source-sink relation. The duration of development stages of flowering, fruit, and tuber development was measured in terms of thermal time. Furthermore, the number of stems, stolons, and tubers was measured, and the yield was registered at the cycle end. The varieties Paola and Primavera reached a greater accumulation of dry matter in each organ and a greater number of tubers by plant about the varieties Paysandú and Violeta. The organs with the greater biomass accumulated were the leaves, stems, and tubers. The source and demand power had a considerable correlation between the NAR and RGR, which increased toward the end of the cycle for Paola and Primavera varieties. The yield and the beginning and duration of the growth stages were related to the season and the varieties.

Phylogenomic discovery of deleterious mutations facilitates hybrid potato breeding

Hybrid potato breeding will transform the crop from a clonally propagated tetraploid to a seed-reproducing diploid. Historical accumulation of deleterious mutations in potato genomes has hindered the development of elite inbred lines and hybrids. Utilizing a whole-genome phylogeny of 92 Solanaceae and its sister clade species, we employ an evolutionary strategy to identify deleterious mutations. The deep phylogeny reveals the genome-wide landscape of highly constrained sites, comprising ∼2.4% of the genome. Based on a diploid potato diversity panel, we infer 367,499 deleterious variants, of which 50% occur at non-coding and 15% at synonymous sites. Counterintuitively, diploid lines with relatively high homozygous deleterious burden can be better starting material for inbred-line development, despite showing less vigorous growth. Inclusion of inferred deleterious mutations increases genomic-prediction accuracy for yield by 24.7%. Our study generates insights into the genome-wide incidence and properties of deleterious mutations and their far-reaching consequences for breeding.

Genomic features of meiotic crossovers in diploid potato.

Meiotic recombination plays an important role in genome evolution and crop improvement. Potato (Solanum tuberosum L.) is the most important tuber crop in the world, but research about meiotic recombination in potato is limited. Here, we resequenced 2163F2 clones derived from five different genetic backgrounds and identified 41 945 meiotic crossovers. Some recombination suppression in euchromatin regions was associated with large structural variants. We also detected five shared crossover hotspots. The number of crossovers in each F2 individual from the accession Upotato 1 varied from 9 to 27, with an average of 15.5, 78.25% of which were mapped within 5kb of their presumed location. We show that 57.1% of the crossovers occurred in gene regions, with poly-A/T, poly-AG, AT-rich, and CCN repeats enriched in the crossover intervals. The recombination rate is positively related with gene density, SNP density, Class II transposon, and negatively related with GC density, repeat sequence density and Class I transposon. This study deepens our understanding of meiotic crossovers in potato and provides useful information for diploid potato breeding.

Using haplotype and QTL analysis to fix favorable alleles in diploid potato breeding.

At present, the potato (Solanum tuberosum L.) of international commerce is autotetraploid, and the complexity of this genetic system creates limitations for breeding. Diploid potato breeding has long been used for population improvement, and because of an improved understanding of the genetics of gametophytic self-incompatibility, there is now sustained interest in the development of uniform F1 hybrid varieties based on inbred parents. We report here on the use of haplotype and quantitative trait locus (QTL) analysis in a modified backcrossing (BC) scheme, using primary dihaploids of S. tuberosum as the recurrent parental background. In Cycle 1, we selected XD3-36, a self-fertile F2 individual homozygous for the self-compatibility gene Sli (S-locus inhibitor). Signatures of gametic and zygotic selection were observed at multiple loci in the F2 generation, including Sli. In the BC1 cycle, an F1 population derived from XD3-36 showed a bimodal response for vine maturity, which led to the identification of late versus early alleles in XD3-36 for the gene CDF1 (Cycling DOF Factor 1). Greenhouse phenotypes and haplotype analysis were used to select a vigorous and self-fertile F2 individual with 43% homozygosity, including for Sli and the early-maturing allele CDF1.3. Partially inbred lines from the BC1 and BC2 cycles have been used to initiate new cycles of selection, with the goal of reaching higher homozygosity while maintaining plant vigor, fertility, and yield.

Potato root-associated microbiomes adapt to combined water and nutrient limitation and have a plant genotype-specific role for plant stress mitigation

BackgroundDue to climate change and reduced use of fertilizers combined stress scenarios are becoming increasingly frequent in crop production. In a field experiment we tested the effect of combined water and phosphorus limitation on the growth performance and plant traits of eight tetraploid and two diploid potato varieties as well as on root-associated microbiome diversity and functional potential. Microbiome and metagenome analysis targeted the diversity and potential functions of prokaryotes, fungi, plasmids, and bacteriophages and was linked to plant traits like tuber yield or timing of canopy closure.ResultsThe different potato genotypes responded differently to the combined stress and hosted distinct microbiota in the rhizosphere and the root endosphere. Proximity to the root, stress and potato genotype had significant effects on bacteria, whereas fungi were only mildly affected. To address the involvement of microbial functions, we investigated well and poorly performing potato genotypes (Stirling and Desirée, respectively) under stress conditions and executed a metagenome analysis of rhizosphere microbiomes subjected to stress and no stress conditions. Functions like ROS detoxification, aromatic amino acid and terpene metabolism were enriched and in synchrony with the metabolism of stressed plants. In Desirée, Pseudonocardiales had the genetic potential to take up assimilates produced in the fast-growing canopy and to reduce plant stress-sensing by degrading ethylene, but overall yield losses were high. In Stirling, Xanthomonadales had the genetic potential to reduce oxidative stress and to produce biofilms, potentially around roots. Biofilm formation could be involved in drought resilience and nutrient accessibility of Stirling and explain the recorded low yield losses. In the rhizosphere exposed to combined stress, the relative abundance of plasmids was reduced, and the diversity of phages was enriched. Moreover, mobile elements like plasmids and phages were affected by combined stresses in a genotype-specific manner.ConclusionOur study gives new insights into the interconnectedness of root-associated microbiota and plant stress responses in the field. Functional genes in the metagenome, phylogenetic composition and mobile elements play a role in potato stress adaption. In a poor and a well performing potato genotype grown under stress conditions, distinct functional genes pinpoint to a distinct stress sensing, water availability and compounds in the rhizospheres.

Genome Report: Genome sequence of 1S1, a transformable and highly regenerable diploid potato for use as a model for gene editing and genetic engineering.

Availability of readily transformable germplasm, as well as efficient pipelines for gene discovery are notable bottlenecks in the application of genome editing in potato. To study and introduce traits such as resistance against biotic and abiotic factors, tuber quality traits and self-fertility, model germplasm that is amenable to gene editing and regeneration is needed. Cultivated potato is a heterozygous autotetraploid and its genetic redundancy and complexity makes studying gene function challenging. Genome editing is simpler at the diploid level, with fewer allelic variants to consider. A readily transformable diploid potato would be further complemented by genomic resources that could aid in high throughput functional analysis. The heterozygous Solanum tuberosum Group Phureja clone 1S1 has a high regeneration rate, self-fertility, desirable tuber traits and is amenable to Agrobacterium-mediated transformation. We leveraged its amenability to Agrobacterium-mediated transformation to create a Cas9 constitutively expressing line for use in viral vector-based gene editing. To create a contiguous genome assembly, a homozygous doubled monoploid of 1S1 (DM1S1) was sequenced using 44 Gbp of long reads generated from Oxford Nanopore Technologies (ONT), yielding a 736 Mb assembly that encoded 31,145 protein-coding genes. The final assembly for DM1S1 represents a nearly complete genic space, shown by the presence of 99.6% of the genes in the Benchmarking Universal Single Copy Orthologs (BUSCO) set. Variant analysis with Illumina reads from 1S1 was used to deduce its alternate haplotype. These genetic and genomic resources provide a toolkit for applications of genome editing in both basic and applied research of potato.

Development of homozygous tetraploid potato and whole genome doubling-induced the enrichment of H3K27ac and potentially enhanced resistance to cold-induced sweetening in tubers.

Polyploid plants typically display advantages on some agronomically important traits over their diploid counterparts. Extensive studies have shown genetic, transcriptomic, and epigenetic dynamics upon polyploidization in multiple plant species. However, few studies have unveiled those alternations imposed only by ploidy level, without any interference from heterozygosity. Cultivated potato is highly heterozygous. Thus, in this study, we developed two homozygous autotetraploid lines and one homozygous diploid line in parallel from a homozygous diploid potato. We confirmed their ploidy levels using chloroplast counting and karyotyping. Oligo-FISH and genome re-sequencing validated that these potato lines are nearly homozygous. We investigated variations in phenotypes, transcription, and histone modifications between two ploidies. Both autotetraploid lines produced larger but fewer tubers than the diploid line. Interestingly, each autotetraploid line displayed ploidy-related differential expression for various genes. We also discovered a genome-wide enrichment of H3K27ac in genic regions upon whole-genome doubling (WGD). However, such enrichment was not associated with the differential gene expression between two ploidies. The tetraploid lines may exhibit better resistance to cold-induced sweetening (CIS) than the diploid line in tubers, potentially regulated through the expression of CIS-related key genes, which seems to be associated with the levels of H3K4me3 in cold-stored tubers. These findings will help to understand the impacts of autotetraploidization on dynamics of phenotypes, transcription, and histone modifications, as well as on CIS-related genes in response to cold storage.

Glycoalkaloid Composition and Flavonoid Content as Driving Forces of Phytotoxicity in Diploid Potato.

Despite their advantages, biotechnological and omic techniques have not been applied often to characterize phytotoxicity in depth. Here, we show the distribution of phytotoxicity and glycoalkaloid content in a diploid potato population and try to clarify the source of variability of phytotoxicity among plants whose leaf extracts have a high glycoalkaloid content against the test plant species, mustard. Six glycoalkaloids were recognized in the potato leaf extracts: solasonine, solamargine, α-solanine, α-chaconine, leptinine I, and leptine II. The glycoalkaloid profiles of the progeny of the group with high phytotoxicity differed from those of the progeny of the group with low phytotoxicity, which stimulated mustard growth. RNA sequencing analysis revealed that the upregulated flavonol synthase/flavonone 3-hydroxylase-like gene was expressed in the progeny of the low phytotoxicity group, stimulating plant growth. We concluded that the metabolic shift among potato progeny may be a source of different physiological responses in mustard. The composition of glycoalkaloids, rather than the total glycoalkaloid content itself, in potato leaf extracts, may be a driving force of phytotoxicity. We suggest that, in addition to glycoalkaloids, other metabolites may shape phytotoxicity, and we assume that these metabolites may be flavonoids.

CRISPR/Cas9-mediated genome editing in diploid and tetraploid potatoes

CRISPR/Cas9-mediated genome editing in diploid and tetraploid potatoes

Generation of Asynaptic Mutants in Potato by Disrupting StDMC1 Gene Using RNA Interference Approach.

Fixing the genomic composition and multiplication through true potato seed (TPS) is an important challenge in autotetraploid potato. Disrupted meiotic cDNA (DMC1) is a meiotic gene that plays a central role in DNA recombination through crossing over in meiosis. Using the Arabidopsis DMC1 (AtDMC1) gene sequence, we retrieved Solanum tuberosum DMC1(StDMC1) from the diploid potato genome, and subsequently, sense and antisense regions of the StDMC1 gene were amplified in potato cv. Kufri Jyoti. The sense and antisense fragments were confirmed by Sanger-sequencing and cloned in the pRI101 vector. Agrobacterium-mediated transformation of the RNAi construct resulted in 44% transformation efficiency, and a total of 137 mutant lines were obtained. These mutant lines were further validated through pollen viability testing, and selected lines were used for gene expression analysis. The acetocarmine-based pollen staining showed reduced pollen viability ranging from 14 to 21% in four DMC1 mutant lines (DMC4-37, DMC4-41, DMC6-20, and DMC6-21), as compared to the Kufri Jyoti control plants, which on average exhibited 78% pollen viability. The phenotypic data was supported by the reduced expression of the StDMC1 gene in these four mutant lines compared to the control Kufri Jyoti. The results confirmed the generation of StDMC1 knockdown lines. This is the first report of StDMC1 mutant line generation in tetraploid potatoes and will be a step forward in generating non-recombinant mutants through sexual reproduction in potatoes.