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AimsDrought is a climate threat limiting crop production. Potato is one of the four most important food crops worldwide and is sensitive to water shortage. The CBP80 gene was shown to affect plant response to drought by regulating the level of microRNA159, and, consequently, the levels of the MYB33 and MYB101 transcription factors. (TF) Our studies aimed to show whether indeed the level of MYB33, MYB65, and MYB101 TFs affects plant response to water shortage.MethodsArabidopsis transgenic plants exhibiting downregulation and Arabidopsis and potato transgenic plants exhibiting overexpression of selected MYB TFs were obtained. Plants response to drought was mainly measured using relative water content (RWC) and stomata closure upon exogenous ABA.ResultsThree MYB TFs studied are involved in plant response to drought. When downregulated in Arabidopsis, the MYB33, MYB65 and MYB101 genes cause stomatal hyposensitivity to ABA, leading to reduced tolerance to drought. Transgenic Arabidopsis and potato plants overexpressing a mutated version of these genes with changed miR159 recognition site, show hypersensitivity to ABA and relatively high tolerance to drought conditions.ConclusionsThe MYB33, MYB65, and MYB101 genes are good be potential targets for innovative breeding to obtain crops with relatively high tolerance to drought.

Soft rot is a bacterial disease that causes heavy losses in potato production worldwide. The goal of this study was to identify quantitative trait loci (QTLs) for potato tuber resistance to bacterium Dickeya solani and for tuber starch content to study the relationship between these traits. A highly resistant diploid hybrid of potato was crossed with a susceptible hybrid to generate the F1 mapping population. Tubers that were wound-inoculated with bacteria were evaluated for disease severity expressed as the mean weight of rotted tubers, and disease incidence measured as the proportion of rotten tubers. Diversity array technology (DArTseq™) was used for genetic map construction and QTLs analysis. The most prominent QTLs for disease severity and incidence were identified in overlapping regions on potato chromosome IV and explained 22.4% and 22.9% of the phenotypic variance, respectively. The second QTL for disease severity was mapped to chromosome II and explained 16.5% of the variance. QTLs for starch content were detected on chromosomes III, V, VI, VII, VIII, IX, XI, and XII in regions different than the QTLs for soft rot resistance. Two strong and reproducible QTLs for resistance to Dickeya solani on potato chromosomes IV and II might be useful for further study of candidate genes and marker development in potato breeding programs. The relationship between tuber resistance to bacteria and the starch content in potato tubers was not confirmed by QTL mapping, which makes the selection of genotypes highly resistant to soft rot with a desirable starch content feasible.

Main conclusion Tuber-omics in potato with the T- and D-types of cytoplasm showed different sets of differentially expressed genes and proteins in response to cold storage. Abstract For the first time, we report differences in gene and protein expression in potato (Solanum tuberosum L.) tubers possessing the T- or D-type cytoplasm. Two F1 diploid reciprocal populations, referred to as T and D, were used. The pooling strategy was applied for detection of differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) in tubers consisting of extreme chip colour after cold storage. RNA and protein bulks were constructed from contrasting phenotypes. We recognized 48 and 15 DEGs for the T and D progenies, respectively. DEPs were identified in the amyloplast and mitochondrial fractions. In the T-type cytoplasm, only 2 amyloplast-associated and 5 mitochondria-associated DEPs were detected. Of 37 mitochondria-associated DEPs in the D-type cytoplasm, there were 36 downregulated DEPs in the dark chip colour bulks. These findings suggest that T- and D-type of cytoplasm might influence sugar accumulation in cold-stored potato tubers in different ways. We showed that the mt/nucDNA ratio was higher in D-possessing tubers after cold storage than in T progeny. For the D-type cytoplasm, the pt/nucDNA ratio was higher for tubers characterized by dark chip colour than for those with light chip colour. Our findings suggest that T- and D-type cytoplasm might influence sugar accumulation in cold-stored potato tubers in different ways.

Potato (Solanum tuberosum L.) is considered a drought-sensitive crop species, although cultivar-dependent differences in tolerance have been described. The extent of drought-induced tuber yield decreases is considered the main criterion for potato tolerance to drought. In this study, eighteen closely related potato cultivars were subjected to drought stress, and among them, two groups with contrasting drought tolerance phenotypes were distinguished (tolerant: Sebago, Katahdin and Cayuga; susceptible: Sequoia and Carpatin). Photosynthesis is the most important primary metabolic process that determines the yield of potato crops and is also strongly affected by drought. The photosynthetic parameters of these two groups of potato cultivars were assessed by measuring chlorophyll a fluorescence. Drought-induced changes in the examined parameters were observed in both groups of cultivars, but the changes in the drought-sensitive group were relatively much more pronounced. The tolerant and sensitive groups significantly differed in terms of most photosynthetic parameters. Principal component analysis (PCA) revealed differences in the responses of the examined potato cultivars and separated tolerant cultivars from sensitive ones, similar to the criterion of the relative decrease in tuber yields. These results suggest that chlorophyll a fluorescence may serve as a useful tool for estimating the level of tolerance to drought stress in potato.

The objective of this study was to map the quantitative trait loci (QTLs) for chip color after harvest (AH), cold storage (CS) and after reconditioning (RC) in diploid potato and compare them with QTLs for starch-corrected chip color. Chip color traits AH, CS, and RC significantly correlated with tuber starch content (TSC). To limit the effect of starch content, the chip color was corrected for TSC. The QTLs for chip color (AH, CS, and RC) and the starch-corrected chip color determined with the starch content after harvest (SCAH), after cold storage (SCCS) and after reconditioning (SCRC) were compared to assess the extent of the effect of starch and the location of genetic factors underlying this effect on chip color. We detected QTLs for the AH, CS, RC and starch-corrected traits on ten potato chromosomes, confirming the polygenic nature of the traits. The QTLs with the strongest effects were detected on chromosomes I (AH, 0 cM, 11.5% of variance explained), IV (CS, 43.9 cM, 12.7%) and I (RC, 49.7 cM, 14.1%). When starch correction was applied, the QTLs with the strongest effects were revealed on chromosomes VIII (SCAH, 39.3 cM, 10.8% of variance explained), XI (SCCS, 79.5 cM, 10.9%) and IV (SCRC, 43.9 cM, 10.8%). Applying the starch correction changed the landscape of QTLs for chip color, as some QTLs became statistically insignificant, shifted or were refined, and new QTLs were detected for SCAH. The QTLs on chromosomes I and IV were significant for all traits with and without starch correction.

Potato virus Y (PVY) is a major potato (Solanum tuberosum L.) pathogen that causes severe annual crop losses worth billions of dollars worldwide. PVY is transmitted by aphids, and successful control of virus transmission requires the extensive use of environmentally damaging insecticides to reduce vector populations. Rysto , from the wild relative S. stoloniferum, confers extreme resistance (ER) to PVY and related viruses and is a valuable trait that is widely employed in potato resistance breeding programmes. Rysto was previously mapped to a region of potato chromosome XII, but the specific gene has not been identified to date. In this study, we isolated Rysto using resistance gene enrichment sequencing (RenSeq) and PacBio SMRT (Pacific Biosciences single-molecule real-time sequencing). Rysto was found to encode a nucleotide-binding leucine-rich repeat (NLR) protein with an N-terminal TIR domain and was sufficient for PVY perception and ER in transgenic potato plants. Rysto -dependent extreme resistance was temperature-independent and requires EDS1 and NRG1 proteins. Rysto may prove valuable for creating PVY-resistant cultivars of potato and other Solanaceae crops.

This study aims the detection of proteins associated with increased resistance of tubers to necrotrophic bacteria Dickeya solani in tetraploid and diploid potato plants. Comparative analysis of differently expressed proteins in tuber tissue of potato cultivars and diploid interspecific hybrids of Solanum, differing in resistance to Dickeya solani, was performed using nano-liquid chromatography coupled with tandem mass spectrometry (LC-MS-MS/MS). Two highly resistant (Bea and Humalda) and three susceptible (Irys, Katahdin, Ulster Supreme) potato cultivars, and the highly resistant (DG 00–270) and the susceptible (DG 08–305) diploid clones, were studied. Proteins were extracted from wounded potato tubers inoculated with bacteria at an early symptomatic phase of infection and from controls, i.e., intact tubers and wounded mock-inoculated tubers. Data are available via ProteomeXchange with identifier PXD013009. Eight constitutive differentially expressed proteins with fold changes ≥1.9 and q-value ≤0.1 between the resistant and susceptible cultivar groups after D. solani infection were selected. Probable inactive patatin-03-Kuras 1 and the proteinase inhibitor PTI exhibited significantly increased protein abundances after bacterial inoculation in both resistant cultivars compared to the susceptible cultivars. In the diploid clones, only metallocarboxypeptidase and metallocarboxypeptidase-like inhibitors exhibited much higher fold changes following pathogenic invasion (274.4- and 368.6-fold, respectively) than after mock inoculation (165.5- and 130.7-fold, respectively). These results show that different proteins indicating significant fold changes between the resistant and susceptible potato cultivars and diploid clones are induced at an early phase of symptomatic D. solani infection.

We demonstrate temporal-, spatial- and temperature-dependent expression of five host microRNAs and target mRNAs in potato leaves after potato virus Y (PVY) inoculation. A novel gene, Ny-DG, conferring resistance to PVY was mapped on chromosome IX in the diploid potato clone DG 81-68. At 20 °C, the PVYNTN strain was localized in the inoculated leaves without symptoms. At 28 °C, necrotic lesions were observed in both the inoculated leaves and the non-inoculated upper leaves and the virus spread systemically, which indicates overcoming the resistance. At 0.5 h post-inoculation and 3-day post-inoculation (dpi), PVY led to significant changes in the levels of the tested miRNAs and their targets only in a small number of cases. At 6 dpi, in the inoculated leaves at 28 °C, the increased expression of stu-miR162, stu-miR168a and stu-miR482 (PotatoMir1005353123_x15854) promoted the downregulation of their targets DCL1, AGO1-2 and Cc-nbs-lrr, respectively. The expression of stu-miR482 (PotatoMir1005658171_x16170) and stu-miR172e remained unchanged, whereas their targets Gpa2 and TOE3 were downregulated. However, in the inoculated leaves at 20 °C, all five tested miRNAs and their targets showed parallel downregulation. In the non-inoculated upper leaves, changes in gene expression levels, namely downregulation, were only detected for the tested five miRNAs at 28 °C. We conclude that the expression patterns of the tested miRNAs and targets were altered differently not only at different time points post-inoculation but also in the inoculated and upper leaves, and their accumulation levels were related to the type of reaction, which in turn was dependent on temperature.

Background Glycoalkaloids are bioactive compounds that contribute to the defence response of plants against herbivore attack and during pathogenesis. Solanaceous plants, including cultivated and wild potato species, are sources of steroidal glycoalkaloids. Solanum plants differ in the content and composition of glycoalkaloids in organs. In wild and cultivated potato species, more than 50 steroidal glycoalkaloids were recognized. Steroidal glycoalkaloids are recognized as potential allelopathic/phytotoxic compounds that may modify the growth of target plants. There are limited data on the impact of the composition of glycoalkaloids on their phytotoxic potential. Results The presence of α-solasonine and α-solamargine in potato leaf extracts corresponded to the high phytotoxic potential of the extracts. Among the differentially expressed genes between potato leaf bulks with high and low phytotoxic potential, the most upregulated transcripts in sample of high phytotoxic potential were anthocyanin 5-aromatic acyltransferase-like and subtilisin-like protease SBT1.7-transcript variant X2. The most downregulated genes were carbonic anhydrase chloroplastic-like and miraculin-like. An analysis of differentially expressed proteins revealed that the most abundant group of proteins were those related to stress and defence, including glucan endo-1,3-beta-glucosidase acidic isoform, whose expression level was 47.96× higher in potato leaf extract with low phytotoxic. Conclusions The phytotoxic potential of potato leaf extract possessing low glycoalkaloid content is determined by the specific composition of these compounds in leaf extract, where α-solasonine and α-solamargine may play significant roles. Differentially expressed gene and protein profiles did not correspond to the glycoalkaloid biosynthesis pathway in the expression of phytotoxic potential. We cannot exclude the possibility that the phytotoxic potential is influenced by other compounds that act antagonistically or may diminish the glycoalkaloids effect.

Tuber starch content (TSC) is a very important trait in potato (Solanum tuberosum L.). This study is the first to use expression quantitative trait loci (eQTL) mapping of transcript-derived markers for TSC in potato. Thirty-four differentially expressed genes were selected by comparing the RNA-seq data of contrasting bulked segregants. For the 11 candidate genes, we determined their relative expression levels across the segregating diploid potato population using RT-qPCR. We detected 36 eQTL as candidate genes distributed on all twelve potato chromosomes, and nine of them overlapped with QTL for TSC. Peaks for two eQTL, eAGPaseS-a and ePGRCRURSE5, were close to the corresponding loci of the large subunit of ADP-glucose pyrophosphorylase (AGPaseS-a) and the 12S globulin cruciferin gene (PGCRURSE5), respectively. The eQTL peaks for AGPaseS-a and PGRCRURSE5 explained 41.0 and 28.3% of the phenotypic variation at the transcript level. We showed the association of the DNA markers for AGPaseS-a and PGRCRURSE5 with QTL for TSC, and significant correlation between the expression level of PGRCRURSE5 and TSC. We did not observe a significant correlation between the expression level of AGPaseS-a and TSC. We concluded that the cruciferin gene PGRCRURSE5 is a novel candidate involved in the regulation of starch content in potato tubers.

Potato virus Y (PVY) is a major potato pathogen that causes annual losses of billions of dollars. Control of its transmission requires extensive use of environmentally damaging insecticides. Rysto confers extreme resistance (ER) to PVY and is a valuable trait in resistance breeding programs. We isolated Rysto using Resistance gene enrichment sequencing (RenSeq) and PacBio SMRT (Pacific Biosciences Single-Molecule Real Time Sequencing). Rysto encodes a nucleotide binding-leucine rich repeat (NLR) protein with an N-terminal TIR domain, and is sufficient for PVY perception and extreme resistance in transgenic potato plants. We investigated the requirements for Rysto-dependent extreme resistance, and showed that Rysto function is temperature-independent and requires EDS1 and NRG1 proteins. Rysto may prove valuable for creating PVY-resistant cultivars of potato and other Solanaceae crops.

The Ny-1 gene confers hypersensitive response (HR) to Potato virus Y (PVY) in potato cultivars (Solanum tuberosum L.). Tetraploid potato breeding clone PB07–37 possessing the allele Ny-1 in a duplex state was developed. After PVY infection, the size of necrotic lesions in leaves of PB07–37 was reduced by approximately 68% in relation to cultivar Rywal plants displaying Ny-1 in a simplex dosage. Two-dimensional gel electrophoresis (2-DE) was applied for screening PVY-induced proteins in leaves of PB07–37. Compared with non-infected control plants, 60 reproducible PVY-induced proteins were detected using LC-MS/MS analysis, of which 41 were involved in qualitative changes and 19 were differently expressed in inoculated leaves by at least 1.5-fold. Proteins involved in the category of photosynthesis and primary metabolism were the most abundant. The results from PB07–37 (Ny-1 duplex) were compared with data from Rywal (Ny-1 simplex). The protein profiles in the Ny-1 simplex and Ny-1 duplex plants are genotype specific. Only eight proteins were identified in both genotypes. Five of them: ATP synthase CF1 alpha chain, chloroplastic; ATP synthase CF1 beta subunit, chloroplastic; ATP synthase subunit beta, mitochondrial-like; linoleate 13S-lipoxygenase 2–1, chloroplastic; and mitochondrial monodehydroascorbate reductase 5 are known to be involved in the defense response in plants. In our study, however, protein abundance did not correspond to the Ny-1 resistance gene dosage.

The Ny-1 and Ry-fsto genes of potato (Solanum tuberosum L.) confer hypersensitive response (HR) and extreme resistance (ER), respectively, to Potato virus Y (PVY). ER-type resistance was also observed in potato plants with both alleles (Ny-1 and Ry-fsto). Using two-dimensional electrophoresis (2-DE), quantitative differences between PVY-infected and non-infected control plants were found for 35, 32 and 15 protein spots identified in leaves of potato cultivar Rywal (possessing Ny-1), and potato tetraploid clones PW 363 (with Ry-fsto) and PB 08–137 (with Ny-1 + Ry-fsto), respectively. We recognized 29, 12 and 21 PVY-induced protein spots involved in qualitative changes in Rywal, PW 363 and PB 08–137 plants, respectively, which were processed and analysed using a liquid chromatography-tandem mass spectrometry (LC-MS/MS) system. A database search indicated that these 62 proteins belong to various functional categories. Their expression was genotype-specific. In the case of cultivar Rywal with HR-mediated resistance, proteins involved in photosynthesis and primary metabolism were the most abundant. For PW 363 and PB 08–137, both with ER–mediated resistance, stress-responsive proteins were the most numerous. Only two proteins – glutamate–glyoxylate aminotransferase 2 (GGAT2) and monodehydroascorbate reductase 5 (MDHAR5) – were identified in all three genotypes. These two proteins are components of the reactive oxygen species (ROS) defence mechanism against pathogens in plants. The present study showed that the differences in PVY-induced proteins in the leaves of Ny-1, Ry-fsto and Ny-1 + R-fsto genotypes do not correspond to the type of gene conferring the resistance or to the observed phenotype.

We report the first comparative study of protein expression profiles in tuber sprouts between Katahdin-derived potato cultivars resistant and susceptible to Synchytrium endobioticum. Synchytrium endobioticum causes wart disease in potato (Solanum tuberosum L.) and is considered as the most important quarantine pathogen in almost all countries where potatoes are grown. We performed a comparative analysis of differentially expressed proteins in the tuber sprouts of potato cultivars differing in resistance to pathotype 1(D1) of S. endobioticum using two-dimensional electrophoresis (2-DE) and liquid chromatography–tandem mass spectrometry (LC–MS/MS) approaches. Bulks prepared from two resistant (Calrose and Humalda) and three susceptible (Sebago, Seneca and Wauseon) potato cultivars were studied. When protein profiles were compared between mock- and S. endobioticum-inoculated sprouts, 35 and 63 protein spots, indicating qualitative or quantitative differences, were detected in the resistant and susceptible cultivars, respectively. In turn, 24 proteins associated with resistance to S. endobioticum were revealed by comparison of the resistant and susceptible bulks. These proteins were changed in a constitutive or induced manner and were grouped into four categories: stress and defence, cell structure, protein turnover, and metabolism. Among the 13 proteins classified into the stress and defence group, seven proteins were related to heat-shock proteins (HSPs)/chaperone factors. In addition, four proteins, S-adenosyl-l-homocysteine hydrolase-like, superoxide dismutase [Mn], inactive patatin-3-Kuras 1 and patatin-15, were induced in the resistant bulk; whereas two proteins, patatin-01 and nucleoredoxin 1, showed significant differences in expression between the S. endobioticum-inoculated resistant and susceptible bulks. The detection of such a large number of S. endobioticum-mediated proteins representing the HSP70, HSP60 and HSP20 families suggests their significant role in restricting wart disease in potato tubers.

The phytotoxic abilities of potato (Solanum tuberosum L.), in a population consisting of wild species, potato diploid hybrids and potato cultivars, against mustard (Sinapis alba L.) were investigated. Measurements of total glycoalkaloid, phenolic and flavonoid concentrations in potato leaf extracts and their role in phytotoxicity were performed. Extracts prepared from wild potato species, particularly Solanum maglia and Solanum chacoense, had the strongest inhibitory effect on mustard growth. Most extracts significantly extended the germination time, as determined by Piepper’s index, but extracts prepared from potato cultivars did not affect seedling growth/development. However, only S. maglia and S. chacoense prolonged the time of germination, corresponding to inhibition of mustard growth. The Pearson’s correlation coefficient of concentrations of total glycoalkaloids, phenolics and flavonoids in potato leaf extracts and length of mustard seedlings showed a significant negative correlation for glycoalkaloids (r = − 0.51), with the strongest effect for root length of potato wild species (r = − 0.79) and potato diploid hybrids (r = − 0.86). Our data indicated that glycoalkaloids are the major phytotoxic compounds responsible for potato phytotoxicity against mustard. However, we observed phenolics and flavonoids may influence the strength of the phytotoxic effects caused by glycoalkaloids.

Key message Most QTL for leaf sucrose content map to positions that are similar to positions of QTL for tuber starch content in diploid potato. Abstract In the present study, using a diploid potato mapping population and Diversity Array Technology (DArT) markers, we identified twelve quantitative trait loci (QTL) for tuber starch content on seven potato chromosomes: I, II, III, VIII, X, XI, and XII. The most important QTL spanned a wide region of chromosome I (42.0–104.6 cM) with peaks at 63 and 84 cM which explained 17.6 and 19.2 % of the phenotypic variation, respectively. ADP-glucose pyrophosphorylase (AGPase) is the key enzyme for starch biosynthesis. The gene encoding the large subunit of this enzyme, AGPaseS-a, was localized to chromosome I at 102.3 cM and accounted for 15.2 % of the variance in tuber starch content. A more than 100-fold higher expression of this gene was observed in RT-qPCR assay in plants with the marker allele AGPaseS-a1334. This study is the first to report QTL for sucrose content in potato leaves. QTL for sucrose content in leaves were located on eight potato chromosomes: I, II, III, V, VIII, IX, X and XII. In 5-week-old plants, only one QTL for leaf sucrose content was detected after 8 h of darkness; four QTL were detected after 8 h of illumination. In 11-week-old plants, 6 and 3 QTL were identified after dark and light phases, respectively. Of fourteen QTL for leaf sucrose content, eleven mapped to positions that were similar to QTL for tuber starch content. These results provide genetic information for further research examining the relationships between metabolic carbon molecule sources and sinks in potato plants. Electronic supplementary material The online version of this article (doi:10.1007/s00122-015-2615-9) contains supplementary material, which is available to authorized users.

Potato virus Y (PVY, Potyvirus) is the fifth most important plant virus worldwide in terms of economic and scientific impact. It infects members of the family Solanaceae and causes losses in potato, tomato, tobacco, pepper and petunia production. In potato and its wild relatives, two types of resistance genes against PVY have been identified. While Ry genes confer symptomless extreme resistance, Ny genes cause a hypersensitive response visible as local necrosis that may also be able to prevent the virus from spreading under certain environmental conditions. The potato cultivar Sarpo Mira originates from Hungary and is highly resistant to PVY, although the source of this resistance remains unknown. We show that cv. Sarpo Mira reacts with a hypersensitive response leading to necrosis after PVYNTN infection in detached leaf, whole plant and grafting assays. The hypersensitivity to PVYNTN segregated amongst 140 individuals of tetraploid progeny of cvs. Sarpo Mira × Maris Piper in a 1:1 ratio, indicating that it was conferred by a single, dominant gene in simplex. Moreover, we identified five DNA markers linked to this trait and located the underlying locus (Ny-Smira) to the long arm of potato chromosome IX. This position corresponds to the location of the Ry chc and Ny-1 genes for PVY resistance. A simple PCR marker, located 1 cM from the Ny-Smira gene, can be recommended for selection of PVY-resistant progeny of cv. Sarpo Mira.

Fusarium oxysporum Schlecht. f.sp. radicis-lycopersici Jarvis & Schoemaker (FORL) is a saprophytic fungus, responsible for the fusarium crown and root rot disease in tomato ( Solanum lycopersicum L. ). This is one of the most destructive pathogens of this species. A new cleaved amplified polymorphic sequence (CAPS) marker C2-25 was developed for the detection of the dominant gene Frl, which confers tomato resistance to FORL . C2-25 was amplified from a conserved ortholog set II (COSII) sequence C2_At2g38025 . The Xap I-derived restriction product of 700 bp was informative for the identification of FORL resistant tomato genotypes and can be used as a diagnostic marker in tomato breeding programmes and hybrid seed production.

Developing new strategies for crop plants to respond to drought is crucial for their innovative breeding. The down-regulation of nuclear cap-binding proteins in Arabidopsis renders plants drought tolerant. The CBP80 gene in the potato cultivar Desiree was silenced using artificial microRNAs. Transgenic plants displayed a higher tolerance to drought, ABA-hypersensitive stomatal closing, an increase in leaf stomata and trichome density, and compact cuticle structures with a lower number of microchannels. These findings were correlated with a higher tolerance to water stress. The level of miR159 was decreased, and the levels of its target mRNAs MYB33 and MYB101 increased in the transgenic plants subjected to drought. Similar trends were observed in an Arabidopsis cbp80 mutant. The evolutionary conservation of CBP80, a gene that plays a role in the response to drought, suggests that it is a candidate for genetic manipulations that aim to obtain improved water-deficit tolerance of crop plants.

Potato (Solanum tuberosum L.) tubers exhibit significant variation in reducing sugar content directly after harvest, cold storage and reconditioning. Here, we performed QTL analysis for chip color, which is strongly influenced by reducing sugar content, in a diploid potato mapping population. Two QTL on chromosomes I and VI were detected for chip color after harvest and reconditioning. Only one region on chromosome VI was linked with cold-induced sweetening. Using the RT-PCR technique, we showed differential expression of the auxin-regulated protein (AuxRP) gene. The AuxRP transcript was presented in light chip color parental clone DG 97-952 and the RNA progeny of the bulk sample consisting of light chip color phenotypes after cold storage. This amplicon was absent in dark chip parental clone DG 08-26/39 and the RNA bulk sample of dark chip progeny. Genetic variation of AuxRP explained up to 16.6 and 15.2 % of the phenotypic variance after harvest and 3 months of storage at 4 °C, respectively. Using an alternative approach, the RDA-cDNA method was used to recognize 25 gene sequences, of which 11 could be assigned to potato chromosome VI. One of these genes, Heat-shock protein 90 (Hsp90), demonstrated higher mRNA and protein expression in RT-qPCR and western blotting assays in the dark chip color progeny bulk sample compared with the light chip color progeny bulk sample. Our study, for the first time, suggests that the AuxRP and Hsp90 genes are novel candidate genes capable of influencing the chip color of potato tubers. Electronic supplementary material The online version of this article (doi:10.1007/s11032-015-0415-1) contains supplementary material, which is available to authorized users.

Drought tolerance in plants is a complex trait involving morphological, physiological, and biochemical mechanisms. Hundreds of genes underlie the response of plants to the stress. For crops, selecting cultivars that can produce economically significant yields under drought is a priority. Potato (Solanum tuberosum L.) is considered as drought sensitive crop, although cultivar-dependent differences in tolerance have been described. Cultivar 'Katahdin' possesses many appropriate characteristics and is widely used for breeding purposes worldwide; it also has enhanced tolerance to drought stress. In this study, we evaluated cv. 'Katahdin' and a half-sib family of 17 Katahdin-derived cultivars for leaf relative water content (RWC) and tuber yield under drought stress. The yields of cultivars 'Wauseon', 'Katahdin', 'Magura', 'Calrose', and 'Cayuga' did not significantly decline under drought stress. Among these five, Wauseon exhibited the lowest reduction in both tuber yield and relative water content under water shortage. The data showed that 'Wauseon' is the most attractive cultivar for studies of molecular and physiological processes under drought and for potato breeding due to low yield losses that correspond with high RWC values. This cultivar can serve as a reservoir of potentially useful genes to develop cultivars with enhanced tolerance to this abiotic stress.

Annexins are a family of calcium- and membrane-binding proteins that are important for plant tolerance to adverse environmental conditions. Annexins function to counteract oxidative stress, maintain cell redox homeostasis, and enhance drought tolerance. In the present study, an endogenous annexin, STANN1, was overexpressed to determine whether crop yields could be improved in potato (Solanum tuberosum L.) during drought. Nine potential potato annexins were identified and their expression characterized in response to drought treatment. STANN1 mRNA was constitutively expressed at a high level and drought treatment strongly increased transcription levels. Therefore, STANN1 was selected for overexpression analysis. Under drought conditions, transgenic potato plants ectopically expressing STANN1 were more tolerant to water deficit in the root zone, preserved more water in green tissues, maintained chloroplast functions, and had higher accumulation of chlorophyll b and xanthophylls (especially zeaxanthin) than wild type (WT). Drought-induced reductions in the maximum efficiency and the electron transport rate of photosystem II (PSII), as well as the quantum yield of photosynthesis, were less pronounced in transgenic plants overexpressing STANN1 than in the WT. This conferred more efficient non-photochemical energy dissipation in the outer antennae of PSII and probably more efficient protection of reaction centers against photooxidative damage in transgenic plants under drought conditions. Consequently, these plants were able to maintain effective photosynthesis during drought, which resulted in greater productivity than WT plants despite water scarcity. Although the mechanisms underlying this stress protection are not yet clear, annexin-mediated photoprotection is probably linked to protection against light-induced oxidative stress.

Despite the long history of breeding potatoes resistant to Phytophthora infestans, this oomycete is still economically the most important pathogen of potato worldwide. The correlation of high levels of resistance to late blight with a long vegetation period is one of the bottlenecks for progress in breeding resistant cultivars of various maturity types. Solanum phureja was identified as a source of effective late blight resistance, which was transferred to the cultivated gene pool by interspecific crosses with dihaploids of Solanum tuberosum. A novel major resistance locus, Rpi-phu1, derived most likely from S. phureja and conferring broad-spectrum resistance to late blight, was mapped to potato chromosome IX, 6.4 cM proximal to the marker GP94. Rpi-phu1 was highly effective in detached leaflet, tuber slice and whole tuber tests during 5 years of quantitative phenotypic assessment. The resistance did not show significant correlation with vegetation period length. Our findings provide a well-characterized new source of resistance for breeding early and resistant-to-P. infestans potatoes.

A novel locus for extreme resistance to Potato virus Y (PVY), Ry-fsto, was identified on potato chromosome XII. The gene Ry-fsto has been introgressed from the wild potato species Solanum stoloniferum. Inheritance of Ry-fsto in the tetraploid potato population Rysto was consistent with the model of a single, dominant gene. Bulked segregant analysis identified an ISSR (inter-simple sequence repeat) marker UBC 857980 linked to Ry-fsto. This marker mapped to linkage group XII of a reference potato RFLP (restriction fragment length polymorphism) map. Chromosome XII specific RFLP markers were converted into PCR-based STS and CAPS markers and tested for linkage with Ry-fsto in the population Rysto. CAPS marker GP122718 was tightly linked to the resistance gene and was successfully used to identify Polish and German cultivars expressing extreme resistance to PVY. This indicates that the source of Ry-fsto has been widely utilized in various potato breeding programs and can be monitored by a diagnostic marker in marker-assisted selection.

A novel locus for potato resistance to potato leafroll virus (PLRV) was characterized by inheritance studies and molecular mapping. The diploid parental clone DW 91-1187 was resistant to PLRV accumulation in both inoculated plants and their tuber progeny. The resistance to PLRV accumulation present in DW 91-1187 was not transmitted to any F1 offspring when crossed with a PLRV susceptible clone. Instead, one half of the F1 individuals exhibited undetectable amounts of PLRV as determined by ELISA during the primary infection assay, but accumulated PLRV in their tuber progeny plants. The other half was clearly infected both in the inoculated and tuber-born plants. The inheritance of resistance to PLRV accumulation may be explained by a model of two complementary alleles of a single gene ( PLRV.4) or by two complementary genes that are closely linked in repulsion phase. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers linked to the PLRV.4 locus were selected. The two complementary factors were closely linked in coupling phase to the alternative alleles UBC864(600) and UBC864(800) of DNA marker UBC864. These markers may be used for marker-assisted selection of genotypes having both factors for resistance to PLRV accumulation. The PLRV.4 locus was mapped to a central position on linkage group XI of the potato molecular map, where no resistance locus has been mapped previously.

With 1 figure and 1 table Abstract Two genes, AOX1a and β-AmyI, were mapped in diploid potato. AOX1a (alternative oxidase 1a) is the cyanide insensitive component of the electron transport chain in plants induced by environmental stress. The enzyme β-amylase (β-AmyI) is involved in hydrolytic starch breakdown in both photosynthetic and storage organs. AOX1a and β-AmyI are located in the central region on potato chromosome VIII, where other genes for potato tuber quality traits were mapped.

The purpose of the study was to investigate the role of salicylic acid (SA) signalling in Ny-1-mediated hypersensitive resistance (HR) of potato (Solanum tuberosum L.) to Potato virus Y (PVY). The responses of the Ny-1 allele in the Rywal potato cultivar and transgenic NahG-Rywal potato plants that do not accumulate SA were characterized at the cytological, biochemical, transcriptome, and proteome levels. Analysis of noninoculated and inoculated leaves revealed that HR lesions started to develop from 3 d post inoculation and completely restricted the virus spread. At the cytological level, features of programmed cell death in combination with reactive oxygen species burst were observed. In response to PVY infection, SA was synthesized de novo. The lack of SA accumulation in the NahG plants led to the disease phenotype due to unrestricted viral spreading. Grafting experiments show that SA has a critical role in the inhibition of PVY spreading in parenchymal tissue, but not in vascular veins. The whole transcriptome analysis confirmed the central role of SA in orchestrating Ny-1-mediated responses and showed that the absence of SA leads to significant changes at the transcriptome level, including a delay in activation of expression of genes known to par ticipate in defence responses. Moreover, perturbations in the expression of hormonal signalling genes were detected, shown as a switch from SA to jasmonic acid/ethylene signalling. Viral multiplication in the NahG plants was accompanied by downregulation of photosynthesis genes and activation of multiple energy-producing pathways.

The two most important bacterial diseases of cultivated potato, blackleg of stems and tuber soft rot, are caused by Erwinia species. Genetic resistance currently present in cultivars is insufficient to protect the crop. New sources of polygenic resistance to Erwinia carotovora ssp. atroseptica (van Hall) Dye (Eca) have been selected in diploid hybrids originating from intercrossing Solanum tuberosum L with the wild species S. chacoense Bitter and S. yungasense Hawk. One F1 hybrid population derived from these materials was used to locate, on the molecular map of potato, quantitative trait loci (QTL) for resistance of tubers and leaves to Eca. A linkage map was constructed based on AFLP and RFLP markers including three resistance-gene-like (RGL) markers. Clustering of AFLP markers in putative centromeric regions was observed. QTL analysis revealed complex inheritance of resistance to Eca. Genetic factors affecting resistance to Eca were located on all 12 potato chromosomes. Putative QTL for tuber resistance were identified on 10 chromosomes. The QTL with the largest and most reproducible effect on tuber resistance mapped to chromosome I. Effects on leaf resistance were less reproducible than effects on tuber resistance. Putative QTL for leaf resistance were identified on 10 chromosomes. Inheritance of tuber and leaf resistance to Eca was largely independent. Several QTL for resistance to Eca were linked to RGL loci. Four of those QTL mapped to genomic segments that have been shown to contain factors for qualitative and quantitative resistance to different pathogens in potato, tomato {Lycopersicon esculentum Mill.), or tobacco {Nicotiana tabacum L.).

The Ns gene confers resistance of potato to Potato virus S (PVS). Sixteen German and Dutch potato cultivars, all registered in Poland, were found to be susceptible to PVS infection. However, scoring of the cultivars for the presence of the Ns-linked SCAR marker SC811(454) revealed additional amplicons with a similar electrophoretic migration rate as that of SC811(454), which resulted in ambiguous determination of the genotype at the Ns locus. MboI or FokI treatment of the PCR products allowed to detect their Ns-unspecificity in PVS-susceptible potato cultivars.

A simple and robust multiplex PCR approach was developed for detection of the alleles Ry-f sto and Ns conferring resistance of potato to Potato Virus Y (PVY) and Potato Virus S (PVS), respectively. Cleaved amplified polymorphic sequence (CAPS) markers GP122564 linked to Ry-f sto and SC811260 linked to Ns were amplified in one PCR reaction and identified after simultaneous digestion of the amplicons with restriction enzymes EcoRV and MboI. Effectiveness of this procedure for marker-assisted selection was confirmed in 55 potato cultivars.

Potato spindle tuber viroid (PSTVd) was transmitted by Myzus persicae to Physalis floridana from P. floridana plants that also were infected with potato leafroll luteovirus (PLRV), whereas it was not transmitted by aphids from plants infected with PSTVd alone. Dot-blot hybridisation was used to detect PSTVd. The results indicate that PLRV can assist PSTVd in its transmission by M. persicae. Doubly infected, aphid-inoculated P. floridana plants from the previous experiment were used as the source plants in aphid transmission tests to the tomato cv. Rutgers, P. floridana and Datura stramonium. PSTVd was detected in 17 of 30 plants of tomato. The viroid was not detected by dot-blotting in any plant of P. floridana and D. stramonium in this experiment, but it was recovered from some plants by sap inoculation of the Rutgers plants. Treatment with RNase A of PLRV preparations purified from doubly infected plants indicated that PSTVd was encapsidated by PLRV particles.

Functional male sterility in tomato (Solanum lycopersicum L.), controlled by the ps-2 and ps genes can be utilized in the production of F1 hybrid tomato seed. Two novel cleaved amplified polymorphic sequence (CAPS) markers linked to the ps-2 and ps genes were found in tomato. The C4-30 and C2-21 markers were developed based on the conserved ortholog set II (COSII) sequences C2_At3g20020 and C2_At1g65900 located on tomato chromosomes 4 and 2, respectively. The HinfI-derived PCR restriction product C4-3080 was applicable in the detection of functional male sterile plants. In case of the C2-21 marker, a polymorphism was revealed after digestion of the amplicon with restriction enzyme MboI. Specificity of the DNA markers identified was verified by scoring the tomato parental lines, F2 progeny and F1 hybrids, in which maternal lines possessed the ps or ps-2 gene. C4-30 and C2-21 can be used as the diagnostic tools in tomato breeding programs and in F1 hybrid seed production.

A random amplified polymorphic DNA marker OPG17 450 linked to the Ns gene that confers resistance of potato to potato virus S (PVS), was used to develop sequence-characterized amplified region (SCAR) markers. After cloning and sequencing of OPG17 450 new polymerase chain reaction (PCR) primers were designed to generate dominant (SCG17 321 ) and codominant (SCG17 448 ) markers. For SCG17 448 , polymorphism between susceptible and resistant genotypes was recovered after digestion of the marker with the restriction enzyme MunI. In addition to the band corresponding to susceptible' allele that does not contain the MunI cleavage site, two bands of approximately 251 bp and 197 bp were observed in the resistant genotypes. The usefulness of these SCAR markers was verified in diploid potatoes possessing the Ns locus from clone G-LKS 678147/60, and in tetraploid potatoes derived from G-LKS 678147/60 and from clone MPI 65118/3.

Sucrose is a substrate for starch biosynthesis, unloaded symplastically into the developing potato tubers. Sucrose is converted to glucose-6-phosphate in the cytosol and exported into the amyloplasts. Starch may be degraded either hydrolytically or phosphorolytically. Glucose and maltose are products of hydrolytic starch breakdown in the sprouting tubers. Glucose phosphates are products of the phosphorolytic activity, metabolized to glucose and fructose in cold-stored tubers. Development of molecular tools for assaying potato gene function provide opportunities to receive genetic progress in the sugar-starch potato breeding.

To assess the reliability of dot-spot hybridization assay for the Potato Spindle Tuber Viroid, more than 1000 potato samples were cross-tested by biological and hybridization methods. Both methods gave similar results with a range of cultivars. The dot-spot hybridization test was reproducible with formaldehyde-denaturated cell-sap samples stored at room temperature for over 6 months. This finding means that samples can be transported from field stations to a diagnostic laboratory and allows for cross-checking of results between laboratories. The data suggest that dot-spot hybridization is suitable for breeding and quarantine purposes.

Hypersensitive resistance (HR) is an efficient defense strategy in plants that restricts pathogen growth and can be activated during host as well as non-host interactions. HR involves programmed cell death and manifests itself in tissue collapse at the site of pathogen attack. A novel hypersensitivity gene, Ny-1, for resistance to Potato virus Y (PVY) was revealed in potato cultivar Rywal. This is the first gene that confers HR in potato plants both to common and necrotic strains of PVY. The locus Ny-1 mapped on the short arm of potato chromosome IX, where various resistance genes are clustered in Solanaceous genomes. Expression of HR was temperature-dependent in cv. Rywal. Strains PVYO and PVYN, including subgroups PVYNW and PVYNTN, were effectively localized when plants were grown at 20 degrees C. At 28 degrees C, plants were systemically infected but no symptoms were observed. In field trials, PVY was restricted to the inoculated leaves and PVY-free tubers were produced. Therefore, the gene Ny-1 can be useful for potato breeding as an alternative donor of PVY resistance, because it is efficacious in practice-like resistance conferred by Ry genes.

Using the RAPD method and bulked segregant analysis we identified four RAPD markers linked to a dominant gene Ns, responsible for a hypersensitive reaction of potato (Solanum tuberosum L.) to potato virus S (PVS) infection. The markers OPE15 550 , OPJ13 500 , OPG17 450 , and OPH19 900 were found to be closely linked to the Ns gene in diploid potato clones. They are situated at 2.6, 3.3, 4.6 and 6.6 cM from Ns, respectively. As a source of the gene, clone G-LKS 678147/60, which is known to carry Ns transferred from S. tuberosum ssp. andigena was used. These RAPD markers were not amplified in resistant tetraploid clones containing Ns derived from the clone MPI 65 118/3, also having an andigenum origin. This suggests that there may be two separate loci of Ns in the sources identified, or different alleles with the same specificity at a single locus, or that the genetic background of tetraploids tested results in different RAPD amplification patterns.

Phytophthora infestans causes an economically important disease of potato called late blight. The epidemic is controlled chemically but resistant potatoes can become an environment-friendly and financially justified alternative solution. The use of diploid Solanum tuberosum derived from European tetraploid cultivars enabled the introgression of novel genes encoding foliage resistance and tuber resistance from other species into the modern cultivated potato gene pool. This study evaluated the resistance of the obtained hybrids, its quality, expression in leaflets and tubers and its relation to the length of vegetation period. We also identified genetic loci involved in late blight resistance and the length of vegetation period. A family of 156 individuals segregating for resistance to late blight was assessed by three laboratory methods: detached leaflet, tuber slice and whole tuber test, repeatedly over 5 years. Length of vegetation period was estimated by a field test over 2 years. The phenotypic distributions of all traits were close to normal. Using sequence-specific PCR markers of known chromosomal position on the potato genetic map, six quantitative trait loci (QTLs) for resistance and length of vegetation period were identified. The most significant and robust QTL were located on chromosomes III (explaining 17.3% of variance observed in whole tuber tests), IV (15.5% of variance observed in slice tests), X (15.6% of variance observed in leaflet tests) and V (19.9% of variance observed in length of vegetation period). Genetic characterization of these novel resistance sources can be valuable for potato breeders and the knowledge that the most prominent QTLs for resistance and vegetation period length do not overlap in this material is promising with respect to breeding early potatoes resistant to P. infestans.

Two new loci for resistance to potato virus M (PVM), Gm and Rm, have been mapped in potato. The gene Gm was derived from Solanum gourlayi, whereas, Solanum megistacrolobum is the source of the gene Rm. Gm confers resistance to PVM infection after mechanical inoculation. Rm induces a hypersensitive response in potato plants. Two diploid populations segregating for Gm and Rm, bulked segregant analysis (BSA) using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR), and available potato molecular maps were instrumental for mapping the resistance loci. The novel locus Gm was mapped to a central region on potato chromosome IX. The locus Rm was placed on the short arm of chromosome XI, close to the marker loci GP250 and GP283, where a hotspot for monogenic and polygenic resistance to diverse pathogens is located in the potato and tomato genome.

The genetic relationship between twenty-six strains of Agaricus bisporus were analysed by the RAPD (random amplified polymorphic DNA) method. DNA amplification was performed with the use of twelve arbitrary 10-mer primers. Four primers, which gave polymorphic band patterns were chosen for RAPD analysis. In total, they gave 24 distinguishable bands, of which nine were polymorphic. The conducted research showed that there is a great genetic similarity among the examined strains. Low polymorphism of the strains may be a proof of a limited genetic pool used in the cultivation of those strains.

Potato leafroll virus (PLRV) causes one of the most widespread and important virus diseases in potato. Resistance to PLRV is controlled by genetic factors that limit plant infection by viruliferous aphids or virus multiplication and accumulation. Quantitative trait locus (QTL) analysis of resistance to virus accumulation revealed one major and two minor QTL. The major QTL, PLRV.1, mapped to potato chromosome XI in a resistance hotspot containing several genes for qualitative and quantitative resistance to viruses and other potato pathogens. This QTL explained between 50 and 60% of the phenotypic variance. The two minor QTL mapped to chromosomes V and VI. Genes with sequence similarity to the tobacco N gene for resistance to Tobacco mosaic virus were tightly linked to PLRV.1. The cDNA sequence of an N-like gene was used to develop the sequence characterized amplified region (SCAR) marker N1271164 that can assist in the selection of potatoes with resistance to PLRV.