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To develop an in vitro assay method for bacterial wilt resistance in potato, resistant and susceptible standard genotypes were grown in vitro, and different conditions of inoculation with Ralstonia solanacearum phylotype I/biovar 4 were examined. The optimal condition was the inoculation of 6-8 leaf stage plants with a bacterial concentration of 102 CFU ml−1 and an incubation temperature of 28 °C. Evaluation of stem wilting was more reliable than that of leaf wilting. Using this method, nine genotypes with different resistance levels in the field were evaluated. Lower disease indices were obtained for genotypes with high resistance levels in the field, suggesting that this assay is useful for evaluating bacterial wilt resistance in a controlled environment. [ABSTRACT FROM AUTHOR]

Greening is a major cause of quality loss in potato tubers. As underground stems, potato tubers are non-photosynthetic plant organs that lack photosynthetic machinery. However, after light exposure, amyloplasts convert to chloroplasts in tuber peripheral cell layers, which cause the accumulation of the green photosynthetic pigment, chlorophyll. Tuber greening can be impacted by genetic, cultural, physiological and environmental factors including planting depth, tuber physiological age, temperature, atmospheric oxygen levels, and lighting conditions. Numerous studies have been devoted to understand and control this costly defect for the potato industry. This review brings together the available knowledge on light-induced greening, from causes to solutions and suggestions on further research with a focus on identifying the underlying mechanisms of tuber greening. [ABSTRACT FROM AUTHOR]

Andean Potato is a major crop for farmers in the Andes and represents an important gene pool for potato improvement. We present the population structure and genetic diversity of 88 Andigena accessions collected in Northwestern Argentina based on functional markers (25 SSR) distributed along 12 chromosomes. Polymorphic information content ranged from 0.40 to 0.87. A Bayesian approach, a Principal Coordinate Analysis and a Cluster Analysis revealed the presence of: I) a major group containing most of the Andean accessions and II) a smaller group including the out-group cv. Spunta and the sequenced genotype DM. Several group specific alleles were detected. AMOVA showed that 81% of the variability was within each group. Eleven of the SSRs analyzed are linked or within genes reported to regulate traits of nutritional and industrial interest. Additionally, the allelic variant of a photoperiod dependent tuberization regulator gene, StCDF1.2, was exclusively detected in accessions clustered in group II. [ABSTRACT FROM AUTHOR]

Potato tubers must be cold-stored to extend their shelf life and maintain an uninterrupted supply chain for food processors. However, a side-effect of low-temperature storage is manifested in terms of cold-induced sweetening (CIS) of potato tubers, which reduces the processing quality and the commercial value of the end-products. RNA interference (RNAi) technology, whereby transgene-derived small interfering RNAs can trigger the homology-based knockdown of cognate host genes and can initiate gene silencing, has been successfully applied in crop improvement through targeted gene knockout in host plants. In the current study, transgenic potato plants (Solanum tuberosum cv. Désirée) were generated, expressing a 300 bp hairpin loop nucleotide sequence targeting the potato vacuolar invertase gene (VInv), under the constitutive Cauliflower mosaic virus 35S promoter. Tubers collected from transgenic lines showed a significant reduction in reducing sugar content after 180 days of cold storage, without showing any measurable off-target effects on plant morphology and tuberization compared to non-transformed control plants. The cold-stored tubers were further assayed for chip color, which showed a fairly light colored quality in the samples originating from RNAi lines. Together with similar effects seen in previously published experiments involving other potato varieties, the Désirée results described here establish the efficacy of using RNAi for the successful reduction of CIS in potato tubers. [ABSTRACT FROM AUTHOR]

'Harimaru' is a new potato variety bred from a cross between 'Saikai 35' as a female parent and 'Pike' as a male parent. Marker selection was performed for 1,647 seedlings to combine resistance genes to late blight (R1), Potato virus Y (Rychc), Potato virus X (Rx1), and golden cyst nematode (H1). In total, 194 selected clones were evaluated in the field, among which the best clone was officially released as 'Harimaru'. Its yield was slightly lower than the local standard variety, 'May Queen'. However, it produces tasty potatoes, that do not become mushy with long boiling times despite its high starch content. 'Harimaru' may become a local specialty potato and its multiple resistance to potato viruses may allow cultivation using homemade seed tubers from the previous season's crop. [ABSTRACT FROM AUTHOR]

Elite parental lines are more likely to breed fine varieties, but knowledge about elite parents and their genetic backgrounds is limited. In this paper, we investigated the pedigree relationships of potato varieties bred worldwide and in China. Several elite parents were identified, and these parents were more frequently used as parents in breeding programs across different time periods and countries. We next used 2b-RAD, a reduced-representation sequencing method, to genotype the elite parent Mira and 24 of its offspring. These cultivars span 5 generations, making this lineage the longest continuous pedigree among Chinese bred potatoes. A total of 47,314 tetraploid single nucleotide polymorphisms (SNPs) identified by FreeBayes were used to trace the conserved segments of the Mira genome. The conserved segments had identical or similar allele-specific SNPs across the analyzed genotypes. In Mira, 3,788 segments comprising over 10,000 bp, or 20.8% of the genome, were defined as conserved segments. These segments contain genes involved in crucial biological processes that are of special interest to breeders. These regions, which have been conserved across generations of highly selective breeding, may be helpful for further breeding and performing genome-wide breeding by design. [ABSTRACT FROM AUTHOR]

The mechanisms of potato (Solanum tuberosum L.) plants’ tolerance to chloride salinity were investigated in cv. Lugovskoi regionalized in Russia. Regenerated plants were produced in vitro from apical meristem and grown on half-strength Murashige and Skoog medium (0.5 MS) using a hydroponic unit in controlled-climate conditions. At the age of six weeks, the plants were exposed to salt stress (50-150 mM NaCl, 7 days). Plant response to salt stress was estimated by growth parameters (fresh and dry biomass of the aboveground and underground parts of plants, linear dimensions of shoot and root, area of leaf surface, and number of stolons) and physiological characteristics (level of photosynthetic pigments, accumulation of sodium, potassium, and calcium ions in the aboveground and underground parts of plants, content of proline, activity of antioxidant enzymes, plant tissue hydration, osmotic potential, and POL). It was found that, in response to salinity, the plants of potato, cv. Lugovskoi, showed a considerable inhibition of growth processes, reduction in chlorophyll a content, and suppression of stolon formation, which points to a rather low salinity tolerance of the cultivar. At the same time, under weak or moderate salt stress, the plants preserved water homeostasis owing to effective osmoregulation, actively accumulated proline that acted as a stress protector, and showed hardly any signs of oxidative stress. It was assumed that low salt tolerance of this cultivar depends on the inability of its root system to retain sodium ions and ensure selective ion transport to the aboveground part of the plant and on inefficiency of the system of sodium ions’ removal from the cytoplasm of leaf cells and their compartmentalization in the central vacuole with the purpose of reducing their toxic effect. The obtained results may be useful for working out a technique of improving salt tolerance of this cultivar by the methods of molecular genetics. [ABSTRACT FROM AUTHOR]

Solanum tuberosum potato species constitute the bulk of economically and agronomically important potato production. However, S. tuberosum is a drought- and frost-sensitive species that is incapable of acclimating to the cold. Solanum commersonii is a tuber-bearing wild potato species that exhibits greater frost and drought resistance than S. tuberosum . CBF/DREB (C-REPET BINDING FACTOR/DROUGHT RESPONSE ELEMENT BINGING FACTOR) transcription factors play important roles in response to a variety of abiotic stresses, such as cold, drought and salt stresses. To explore different functions between S. tuberosum CBF1 (StCBF1) and S. commersonii CBF1 (ScCBF1), Arabidopsis was transformed with the ScCBF1 and StCBF1 genes driven by a constitutive CaMV35S promoter. Our results reveal that the ScCBF1 transgenic lines are much more tolerant to freezing and drought than the StCBF1 transgenic lines. The development of transgenic plants was altered, resulting in dwarf phenotype with delayed flowering and thicker and additional rosette leaves. The expression levels of several COR (COLD-RESPONSIVE) genes and development-related genes, including genes that inhibited plant growth (GA2ox7, RGL3) and delayed flowering (FLC) were higher in transgenic plants. These results suggest that these two potato CBF1 play important roles in the plant response to abiotic stress and can influence plant growth and development, and ScCBF1 plays a more pronounced function than StCBF1. [ABSTRACT FROM AUTHOR]

Polyploidy is one of the most important events in plant evolution: the evidence suggests that more than 70% of the angiosperms underwent some event of genomic duplication during their diversification. In potato, the third more important crop worldwide, the effects of autopolyploidization at morphological, physiological and molecular levels have been explored and no evident phenotypic superiority of polyploid lines respect to the parental diploid lines has been found. On the other hand, the consequences of allopolyploidization have not been analysed so far in potato from a molecular point of view. In this work we study the variability induced by whole genome duplication in potato auto and allotetraploids obtained by chromosomal duplication of a S. kurtzianum genotype (autopolyploid model) and by chromosomal duplication of a diploid interspecific hybrid between S. tuberosum and S. kurtzianum (allopolyploid model). Polyploid lines in both models showed a tendency to increase the vigor in phenotypic characters compared with diploid parent they derived from. AFLP analysis showed no polymorphism between parental diploid and derived polyploids lines. However, there were DNA methylation changes between diploids and polyploids, indicating that polyploidization alters the epigenetic patterns in potato. It is possible that the superiority of the tetraploid can be explained as a heterotic response dependent on the presence of heterozygous loci and epistatic interactions inducing alterations in the gene regulation of the polyploid genomes. [ABSTRACT FROM AUTHOR]

Abstract: A total of four traits related to bioactive potential of the tetraploid potato were studied in a North Carolina II cross‐experiment. The objectives of this study were to estimate genetic parameters for target bioactive compounds in a tetraploid potato population, to determine phenotypic and genotypic correlations and to calculate combining ability effects of cultivars and breeding lines included in the crossing scheme. Total monomeric anthocyanins, phenolics, carotenoids and hydrophilic antioxidant capacity were measured by spectrophotometric methods. The sources of variation female (GCAf), male (GCAm) and their interaction (SCA) were significant for bioactive compounds and antioxidant capacity. Narrow‐sense heritability estimates (h2) were moderate for all the traits (0.413–0.657). Both phenotypic and genotypic correlations were highly significant for phenolic compounds and antioxidant capacity. As regards combining ability, positive and negative GCA and also SCA effects have been identified for bioactive compounds and antioxidant capacity. These results indicate that hybrid breeding lines can be obtained by choosing parents with significant GCA, whose bioactive potential can also be further enhanced if the crosses have a significant SCA. [ABSTRACT FROM AUTHOR]

In a remote mountainous region of the Philippines, farmers selected their own clones from hybrid True Potato Seed (TPS) populations and have maintained them for 30 years without public support. In 1985, the International Potato Center (CIP) initiated on-farm TPS research in the Mount Kanlaon area to help farmers control or reduce the rates of bacterial wilt caused by Ralstonia solanacearum. Seedlings were either grown in nursery beds or as transplants in the field. At harvest, farmers not only harvested their crop for either food or for sale but also selected their preferred clones. A survey conducted in 2016 showed that farmers are still growing potato clones selected from TPS and that these clones had spread to numerous areas within and around Mount Kanlaon. Farmers kept these clones because it was profitable because they required minimum inputs and their resistance to various pests and diseases and adverse weather conditions. ELISA tests showed that these clones have excellent virus resistance which partially explains why these clones have been growing for 30 years without a formal seed production program or any government support. Parents used to develop the TPS hybrids included those with virus and R. solanacearum resistance. [ABSTRACT FROM AUTHOR]

The golden cyst nematode (Globodera rostochiensis) is a serious pest that can dramatically reduce potato crop yield. Pathotype Ro1 of G. rostochiensis was first detected in the United States in 1941 and is still present on several farms in New York State. The H1 gene confers high levels of resistance to pathotype Ro1 but screening for it with a bioassay is time consuming and expensive. In this study two known molecular markers, 57R and TG689, were evaluated for their ability to identify resistant clones among 38 global cultivars and 350 New York breeding clones. The ability of either marker to predict resistance was high - 99.7% and 98.3% for 57R and TG689, respectively - but the ability to predict susceptibility was much lower, 47% and 41%, respectively. As resistance is the trait of interest, either of these markers is sufficient to make selection decisions in a practical breeding program. Cases exhibiting discordance between presence/absence of diagnostic markers and bioassay results were investigated further. Recombination, inflow of other resistance genes, and occasional failure of marker- and/or bio-assays are discussed as potential causes. [ABSTRACT FROM AUTHOR]

Common scab caused by the soil-borne bacterium Streptomyces scabies is a serious disease for the potato industry. We have identified a strong source of resistance in the diploid wild relative Solanum chacoense. This resistance has been introgressed into tetraploid cultivated potato via unilateral sexual polyploidization. This paper describes three hybrid clones (M8, M16, M17) for use by potato breeding programs to enhance resistance to common scab. They were created by crossing a diploid clone (50% S. tuberosum, 50% S. chacoense) to tetraploid cultivars, producing hybrids with 75% cultivated and 25% wild germplasm. The clones are male and female fertile, and are adapted to temperate zone production environments. [ABSTRACT FROM AUTHOR]

Key message: Cd is a toxic metal, whilst Zn is an essential for plant and human health. Both can accumulate in potato tubers. We examine the genetic control of this process.Abstract: The aim of this study was to map quantitative trait loci (QTLs) influencing tuber concentrations of cadmium (Cd) and zinc (Zn). We developed a segregating population comprising 188 F1 progeny derived from crossing two tetraploid cultivars exhibiting divergent tuber-Cd-accumulation phenotypes. These progeny were genotyped using the SolCap 8303 SNP array, and evaluated for Cd, Zn, and maturity-related traits. Linkage and QTL mapping were performed using TetraploidSNPMap software, which incorporates all allele dosage information. The final genetic map comprised 3755 SNP markers with average marker density of 2.94 per cM. Tuber-Cd and Zn concentrations were measured in the segregating population over 2 years. QTL mapping identified four loci for tuber-Cd concentration on chromosomes 3, 5, 6, and 7, which explained genetic variance ranging from 5 to 33%, and five loci for tuber-Zn concentration on chromosome 1, 3, 5, and, 6 explaining from 5 to 38% of genetic variance. Among the QTL identified for tuber-Cd concentration, three loci coincided with tuber-Zn concentration. The largest effect QTL for both tuber-Cd and Zn concentration coincided with the maturity locus on chromosome 5 where earliness was associated with increased tuber concentration of both metals. Coincident minor-effect QTL for Cd and Zn sharing the same direction of effect was also found on chromosomes 3 and 6, and these were unrelated to maturity The results indicate partially overlapping genetic control of tuber-Cd and Zn concentration in the cross, involving both maturity-related and non-maturity-related mechanisms. [ABSTRACT FROM AUTHOR]

The DWARF4 ( DWF4 ) gene encodes a C-22 hydroxylase which is pivotal for brassinosteroids (BRs) biosynthesis. In this research, aimed to understand the molecular mechanism of DWF4 on regulation of potatoes tolerance to salt stress, DWF4 was cloned from potato, named as StDWF4 . Its 1476 bp open reading frame encodes a protein of 491 amino acids. The StDWF4 -overexpressing (OE) and interference-expressing (RNAi) transgenic potato plants were acquired using Agrobacterium -mediated transformation, respectively. Tissue specific analysis using Quantitative real-time polymerase chain reaction (qRT-PCR) demonstrated that the StDWF4 gene expressed in the leaves, stems and roots of the transgenic and un-transgenic (NT) plants, with specially increased ( StDWF4 -OE)/reduced ( StDWF4 -RNAi) expression in the roots. The content of malondialdehyde (MDA) in StDWF4 -OE potato plants was lower than that of NT, and proline content was higher than that of NT. MDA and proline content in StDWF4 -OE and NT under salt-stress was significantly higher than that of the control and was increased at different sampling times. The content of soluble protein, soluble sugar and the activities of superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX) was higher in the StDWF4 -OE plantlets at varied salt treatment time than in the NT potatoes. Reduction of H 2 O 2 content in the StDWF4 -OE plants was observed. All above plant physiology indicators in the StDWF4 -RNAi potatoes showed opposite variation trends. The results proved that the overexpressing of StDWF4 in potato plantlets can enhance the salt resistance by alleviating the negative effects of salt-stress. However, its interference expression in potato plants depresses the salt resistance. The results lay the groundwork for intensive study of BRs regulation in potato growth and development, and will help us to reveal the molecular mechanisms of how the BRs signaling regulate potato salt tolerance. [ABSTRACT FROM AUTHOR]

The reported narrow genetic base of cultivated potato (Solanum tuberosum) can be expanded by the introgression of many related species with large genetic diversity. The analysis of the genetic structure of a potato population is important to broaden the genetic base of breeding programs by the identification of different genetic pools. A panel composed by 231 diverse genotypes was characterized using single nucleotide polymorphism (SNP) markers of the Illumina Infinium Potato SNP Array V2 to identify population structure and assess genetic diversity using discriminant analysis of principal components (DAPC) and pedigree analysis. Results revealed the presence of five clusters within the populations differentiated principally by ploidy, taxonomy, origin and breeding program. The information obtained in this work could be readily used as a guide for parental introduction in new breeding programs that want to maximize variability by combination of contrasting variability sources such as those presented here. [ABSTRACT FROM AUTHOR]

Relative gene expression analyses by qRT-PCR (quantitative reverse transcription PCR) require an internal control to normalize the expression data of genes of interest and eliminate the unwanted variation introduced by sample preparation. A perfect reference gene should have a constant expression level under all the experimental conditions. However, the same few housekeeping genes selected from the literature or successfully used in previous unrelated experiments are often routinely used in new conditions without proper validation of their stability across treatments. The advent of RNA-Seq and the availability of public datasets for numerous organisms are opening the way to finding better reference genes for expression studies. Globodera rostochiensis is a plant-parasitic nematode that is particularly yield-limiting for potato. The aim of our study was to identify a reliable set of reference genes to study G. rostochiensis gene expression. Gene expression levels from an RNA-Seq database were used to identify putative reference genes and were validated with qRT-PCR analysis. Three genes, GR, PMP-3, and aaRS, were found to be very stable within the experimental conditions of this study and are proposed as reference genes for future work. [ABSTRACT FROM AUTHOR]

Studying phenotypic and genomic modifications associated with pathogen adaptation to resistance is a crucial step to better understand and anticipate resistance breakdown. This short review summarizes recent results obtained using experimentally evolved populations of the potato cyst nematode Globodera pallida. In a first step, the variability of resistance durability was explored in four different potato genotypes carrying the resistance quantitative trait loci (QTL) GpaVvrn originating from Solanum vernei but differing by their genetic background. The consequences of the adaptation to resistance in terms of local adaptation, cross-virulence and virulence cost were then investigated. Finally, a genome scan approach was performed in order to identify the genomic regions involved in this adaptation. Results showed that nematode populations were able to adapt to the QTL GpaVvrn, and that the plant genetic background has a strong impact on resistance durability. A trade-off between the adaptations to different resistant potato genotypes was detected, and we also showed that adaptation to the resistance QTL GpaVvrn from S. vernei did not allow adaptation to the colinear locus from S. sparsipilum (GpaVspl). Unexpectedly, the adaptation to resistance led to an increase of virulent individual’s fitness on a susceptible host. Moreover, the genome scan approach allowed the highlighting of candidate genomic regions involved in adaptation to host plant resistance. This review shows that experimental evolution is an interesting tool to anticipate the adaptation of pathogen populations and could be very useful for identifying durable strategies for resistance deployment. [ABSTRACT FROM AUTHOR]

The activity of the pool of sgRNA molecules designed for different regions of potato coilin and phytoene desaturase genes was compared in vitro. Due to the presence of nucleotides unpaired with DNA, sgRNA is able not only to inhibit but also to stimulate the activity of the Cas9-sgRNA complex in vitro. Although the first six nucleotides located in the DNA substrate proximally to the PAM site at the 3' end are the binding sites for cas9, they had no significant effect on the activity of the Cas9-sgRNA complex. [ABSTRACT FROM AUTHOR]

Transgenic potato plants harbouring cry1Ab gene, resistant to potato tuber moth, were used to examine the persistence of recombinant DNA and proteins in the digesta, gastrointestinal tract (GIT) tissues and visceral organs of rats in a 90-day feeding trial. Native plant DNA (chloroplastic gene, cp) and fragments of cry1Ab, nptII and nos promoter were tracked in rat organs and tissues by polymerase chain reaction (PCR). Although complete sequences (1200 bp) of cry1Ab gene and its decreased size (400 bp) were detected in the rat diets, the presence of these sequences was not confirmed by PCR analysis in the GIT contents of rats fed diets containing transgenic potato. Moreover, the 400 bp sequence of the nptII gene and 300 bp of the nos promoter were detected in the caecum, stomach and rectum contents of some rats fed transgenic potato; however, these sequences were not detected in all rats. Immunoassay showed that Cry1Ab protein is detectable in the GIT contents of rats fed transgenic potato, but quantitative assay by ELISA confirmed that the Cry1Ab protein was partially degraded (80.3-83.7% digestibility) after passing through the GIT. Similarly, ELISA assay of NPTII showed that the protein can be digested (76.8-84.36% digestibility) in the GIT. So, although ingested native and recombinant DNA and protein did not totally degrade in the GIT, it can be indicated that there is no need for concern about the effects of recombinant cry1Ab and nptII genes and their products on animals and humans. [ABSTRACT FROM AUTHOR]

The objective of this case study is to examine the farm management decision of whether to adopt a new, genetically engineered potato variety. We describe the potato supply chain from seed production to final consumer products and explore how price and production risk interact to influence decision making at each link in that chain. We provide extensive supplemental material as well, including a teaching note with assignment and/or discussion questions, an introduction to and application of stakeholder theory, and a tool that assists students in calculating expected and simulated actual returns from their choice of potato variety. [ABSTRACT FROM AUTHOR]

Abstract Chlorogenic acid (CGA) plays an important role in protecting plants against pathogens and promoting human health. Although CGA accumulates to high levels in potato tubers, the key enzyme p -coumaroyl quinate/shikimate 3′-hydroxylase (C3′H) for CGA biosynthesis has not been isolated and functionally characterized in potato. In this work, we cloned StC3′H from potato and showed that it catalyzed the formation of caffeoylshikimate and CGA (caffeoylquinate) from p -coumaroyl shikimate and p -coumaroyl quinate, respectively, but was inactive towards p -coumaric acid in in vitro enzyme assays. When the expression of StC3′H proteins was blocked through antisense (AS) inhibition under the control of a tuber-specific patatin promoter, moderate changes in tuber yield as well as phenolic metabolites in the core tuber tissue were observed for several AS lines. On the other hand, the AS and control potato lines exhibited similar responses to a bacterial pathogen Pectobacterium carotovorum. These results suggest that St C3′H is implicated in phenolic metabolism in potato. They also suggest that CGA accumulation in the core tissue of potato tubers is an intricately controlled process and that additional C3′H activity may also be involved in CGA biosynthesis in potato. Highlights • St C3′H catalyzed the formation of coumaroyl quinate and coumaroyl shikimate in vitro. • Antisense downregulation of St C3′H in potato tubers caused moderate increases in ferulic acid conjugates and chlorogenic acid. • A reduction in tuber yield was observed in antisense StC3′H lines, though their response to a bacterial pathogen was not significantly affected. [ABSTRACT FROM AUTHOR]

Summary: Potato is one of the four most important food crop plants worldwide and is strongly affected by drought. The following two pairs of potato cultivars, which are related in ancestry but show different drought tolerances, were chosen for comparative gene expression studies: Gwiazda/Oberon and Tajfun/Owacja. Comparative RNA‐seq analyses of gene expression differences in the transcriptomes obtained from drought‐tolerant versus drought‐sensitive plants during water shortage conditions were performed. The 23 top‐ranking genes were selected, 22 of which are described here as novel potato drought‐responsive genes. Moreover, all but one of the potato genes selected have homologues in the Arabidopsis genome. Of the seven tested A. thaliana mutants with altered expression of the selected homologous genes, compared to the wild‐type Arabidopsis plants, six showed an improved tolerance to drought. These genes encode carbohydrate transporter, mitogen‐activated protein kinase kinase kinase 15 (MAPKKK15), serine carboxypeptidase‐like 19 protein (SCPL19), armadillo/beta‐catenin‐like repeat‐containing protein, high‐affinity nitrate transporter 2.7 and nonspecific lipid transfer protein type 2 (nsLPT). The evolutionary conservation of the functions of the selected genes in the plant response to drought confirms the importance of these identified potato genes in the ability of plants to cope with water shortage conditions. Knowledge regarding these gene functions can be used to generate potato cultivars that are resistant to unfavourable conditions. The approach used in this work and the obtained results allowed for the identification of new players in the plant response to drought. [ABSTRACT FROM AUTHOR]

Steroidal glycoalkaloids (SGAs), while found in many solanaceous plants, can accumulate to unacceptably high levels in potato tubers. The two primary SGAs that occur in potatoes are the tri-glycosylated alkaloids, α-solanine and α-chaconine. The first glycosylation steps in their biosynthetic pathways are performed by the regulated enzymes SGT1, the UDP-galactose:solanidine galactosyltransferase, and SGT2, the UDP-glucose:solanidine glucosyltransferase, respectively. Using fragments of the Sgt1 and Sgt2 genes to produce small inhibitory RNAs (siRNA), we have been able to down-regulate each branch of the pathway. The use of the siRNA approach increases the efficiency of producing transgenic plant lines with reductions in individual SGAs but further research is required to achieve reductions in levels of total SGA accumulation. [ABSTRACT FROM AUTHOR]

Six hundred untrained panelists evaluated the baked sensory appeal of up to six varieties, three clones, and two Russet Norkotah (RN) strains. Panelists consumed small samples of plain baked potato and recorded their preference for aroma, flavor, texture, aftertaste and acceptance. After viewing photographs of two unidentified baked potatoes (RN and Classic Russet) sliced in half, they selected the photo that best described their visual preference of a baked potato; each potato was stored at 6.7 °C for 6 months prior to cooking. Eighty percent of panelists shown the photographs of the unidentified baked potatoes preferred the visual appearance of Classic R (white/cream - colored flesh) to that of RN (yellow/Gy - colored flesh). Mean scores for all culinary traits averaged > 5.0 on the 1-9 scale, which indicated that panelists generally liked the culinary attributes of all trialed varieties, however, panelists were able to detect differences among varieties. Flavor and texture were addressed in written comments from panelists more frequently than other attributes. On a texture scale of 1-7, with 1 = moist/dense and 7 = dry/crumbly, most panelists preferred baked potatoes with a texture of 3 (creamy/smooth). [ABSTRACT FROM AUTHOR]

Plants have evolved different types of immune reactions but large-scale proteomics about these processes are lacking, especially in the case of agriculturally important crop pathosystems. We have established a system for investigating PAMP-triggered immunity (PTI) and two different effector-triggered immunity (ETI; triggered by Avr2 or IpiO) responses in potato. The ETI responses are triggered by molecules from the agriculturally important Phytophthora infestans interaction. To perform large-scale membrane protein-based comparison of these responses, we established a method to extract proteins from subcellular compartments in leaves. In the membrane fractions that were subjected to quantitative proteomics analysis, we found that most proteins regulated during PTI were also regulated in the same way in ETI. Proteins related to photosynthesis had lower abundance, while proteins related to oxidative and biotic stress, as well as those related to general antimicrobial defense and cell wall degradation, were found to be higher in abundance. On the other hand, we identified a few proteins—for instance, an ABC transporter-like protein—that were only found in the PTI reaction. Furthermore, we also identified proteins that were regulated only in ETI interactions. These included proteins related to GTP binding and heterotrimeric G-protein signaling, as well as those related to phospholipase signaling. [ABSTRACT FROM AUTHOR]

Summary: The evolution of duplicated genes after polyploidization has been the subject of many evolutionary biology studies. Potato (Solanum tuberosum) and tomato (Solanum lycopersicum) are the first two sequenced genomes of asterids, and share a common polyploidization event. However, the epigenetic role of DNA methylation on the evolution of duplicated genes derived from polyploidization is not fully understood. Here, we explore the role of the DNA methylation in the evolution of duplicated genes in potato and tomato. The overall levels of DNA methylation are different, although patterns of DNA methylation are similar in potato and tomato. Different types of duplicated genes can display different methylation patterns in potato and tomato. In addition, we found that differences in the methylation levels between duplicated genes were associated with gene expression divergence. In particular, for the majority of duplicated gene pairs, one copy is always hyper‐ or hypo‐methylated compared with the other copy across different tomato fruit ripening stages, and these genes are enriched for specific function related to transcription factor (TF) activity. Furthermore, transcription of hundreds of duplicated TFs was shown to be regulated by DNA methylation during fruit ripening stages in tomato, some of which are well‐known fruit ripening TFs. Taken together, our results support the notion that DNA methylation may facilitate divergent evolution of duplicated genes and play roles in important biological processes such as tomato fruit ripening. [ABSTRACT FROM AUTHOR]

Background: Recent advances in ~omics technologies such as transcriptomics, metabolomics and proteomics along with genotypic profiling have permitted the genetic dissection of complex traits such as quality traits in non-model species. To get more insight into the genetic factors underlying variation in quality traits related to carbohydrate and starch metabolism and cold sweetening, we determined the protein content and composition in potato tubers using 2D-gel electrophoresis in a diploid potato mapping population. Upon analyzing we made sure that the proteins from the patatin family were excluded to ensure a better representation of the other proteins. Results: We subsequently performed pQTL analyses for all other proteins with a sufficient representation in the population and established a relationship between proteins and 26 potato tuber quality traits (e.g. flesh colour, enzymatic discoloration) by co-localization on the genetic map and a direct correlation study of protein abundances and phenotypic traits. Over 1643 unique protein spots were detected in total over the two harvests. We were able to map pQTLs for over 300 different protein spots some of which co-localized with traits such as starch content and cold sweetening. pQTLs were observed on every chromosome although not evenly distributed over the chromosomes. The largest number of pQTLs was found for chromosome 8 and the lowest for chromosome number 10. For some 20 protein spots multiple QTLs were observed. Conclusions: From this analysis, hotspot areas for protein QTLs were identified on chromosomes three, five, eight and nine. The hotspot on chromosome 3 coincided with a QTL previously identified for total protein content and had more than 23 pQTLs in the region from 70 to 80 cM. Some of the co-localizing protein spots associated with some of the most interesting tuber quality traits were identified, albeit far less than we had anticipated at the onset of the experiments. [ABSTRACT FROM AUTHOR]

Background: Heat shock proteins (Hsps) are essential components in plant tolerance mechanism under various abiotic stresses. Hsp20 is the major family of heat shock proteins, but little of Hsp20 family is known in potato (Solanum tuberosum), which is an important vegetable crop that is thermosensitive. Results: To reveal the mechanisms of potato Hsp20s coping with abiotic stresses, analyses of the potato Hsp20 gene family were conducted using bioinformatics-based methods. In total, 48 putative potato Hsp20 genes (StHsp20s) were identified and named according to their chromosomal locations. A sequence analysis revealed that most StHsp20 genes (89.6%) possessed no, or only one, intron. A phylogenetic analysis indicated that all of the StHsp20 genes, except 10, were grouped into 12 subfamilies. The 48 StHsp20 genes were randomly distributed on 12 chromosomes. Nineteen tandem duplicated StHsp20s and one pair of segmental duplicated genes (StHsp20-15 and StHsp20-48) were identified. A cis-element analysis inferred that StHsp20s, except for StHsp20-41, possessed at least one stress response cis-element. A heatmap of the StHsp20 gene family showed that the genes, except for StHsp20-2 and StHsp20-45, were expressed in various tissues and organs. Real-time quantitative PCR was used to detect the expression level of StHsp20 genes and demonstrated that the genes responded to multiple abiotic stresses, such as heat, salt or drought stress. The relative expression levels of 14 StHsp20 genes (StHsp20-4, 6, 7, 9, 20, 21, 33, 34, 35, 37, 41, 43, 44 and 46) were significantly up-regulated (more than 100-fold) under heat stress. Conclusions: These results provide valuable information for clarifying the evolutionary relationship of the StHsp20 family and in aiding functional characterization of StHsp20 genes in further research. [ABSTRACT FROM AUTHOR]

The objectives of this research were to study tuber starch characteristics and chemical - thermal properties of 21 potato varieties, and to determine their genetic diversity through SSR markers. Starch granular size varied among samples, with a wide diameter distribution (5–85 μm), while granule shapes were similar. Differential Scanning Calorimeter analysis showed that the transition temperatures (69 °C–74 °C) and enthalpies of gelatinization (0.9 J/g–3.8 J/g) of tubers were also variety dependent. SSR analysis allowed the detection of 157 alleles across all varieties, with an average value of 6.8 alleles per locus. Variety-specific alleles were also identified. SSR-based cluster analysis revealed that varieties with interesting quality attributes were distributed among all clusters and sub-clusters, suggesting that the genetic basis of traits analyzed may differ among our varieties. The information obtained in this study may be useful to identify and develop varieties with slowly digestible starch. [ABSTRACT FROM AUTHOR]

Glutathione S-transferases (GSTs) are ubiquitous enzymes which play versatile functions including cellular detoxification and stress tolerance. In this study, a comprehensive genome-wide identification of GST gene family was carried out in potato ( Solanum tuberosum L.). The result demonstrated the presence of at least 90 GST genes in potato which is greater than any other reported species. According to the phylogenetic analyses of Arabidopsis , rice and potato GST members, GSTs could be subdivided into ten different classes and each class is found to be highly conserved. The largest class of potato GST family is tau with 66 members, followed by phi and lambda. The chromosomal localization analysis revealed the highly uneven distribution of StGST genes across the potato genome. Transcript profiling of 55 StGST genes showed the tissue-specific expression for most of the members. Moreover, expression of StGST genes were mainly repressed in response to abiotic stresses, while largely induced in response to biotic and hormonal elicitations. Further analysis of StGST gene's promoter identified the presence of various stress responsive cis-regulatory elements. Moreover, one of the highly stress responsive StGST members, StGSTU46, showed strong affinity towards flurazole with lowest binding energy of − 7.6 kcal/mol that could be used as antidote to protect crop against herbicides. These findings will facilitate the further functional and evolutionary characterization of GST genes in potato. [ABSTRACT FROM AUTHOR]

Potato tubers with purple flesh contain high levels of antioxidants as do some of many berries fruits. This study evaluated the total antioxidant activity (TAC) of four potatoes genotypes with purple colored flesh tubers in comparison with cranberries, blueberries, blackberries and raspberries. The potential antioxidant activity (estimated byABTS method and DPPH scavenging activity) of the purple potato tubers were closed to that obtained for raspberries. Compared to other rich sources of polyphenols like the berries, the purple potato consumption has as special advantages the bioavailability and economic considerations (lower price). [ABSTRACT FROM AUTHOR]

The production of minitubers using the hydroponic method has become an important component in the production of seed potatoes in developed countries. The objective of the study is obtaining minitubers (Prebase), free from diseases, from the transplantation of vitroplants on industrial substrate and different hydroponic systems. The hydroponic culture involves growing plants on artificial substrates, or in a sterile, porous substrate with a high-water permeability. The parameters analyzed in this study were: average number of minitubers / plant and average weight of minitubers / plant. By using hydroponic system, a high minitubes production was obtained. The trifactor experience of the type 4 x 2 x 2 with 16 variants used the following graduations of the studied factors: experimental factor a, variety, with four graduations: a1- Braşovia; a2- Castrum; a3- Marvis; a4- Sarmis; experimental factor b, culture substrate, with two graduations: b1- expanded clay; b2 - perlite; experimental factor c, the hydroponic system: c1- Wilma; c2- Nutrient Film Technique. These variants were experimented in 3 repetitions. Castrum variety was remarked by obtaining a high number of minitubers and also by weight of minitubers. Nutrient Film Technique hydroponic system had a positive influence on minituberization, for minitubers number, but also for weight of minitubers. Perlite substrate has determined obtaining high values for both studied parameters. [ABSTRACT FROM AUTHOR]

Phosphate transporter (PHTs) have important roles in Pi acquisition, allocation, and signal transduction. The aim of this study is to provide a comprehensive knowledge of PHTs in potato. Very strict homology search and subsequent domain verification using Hidden Markov Models revealed that the potato genome has 20 StPHT and StPHOs genes which were grouped into 5 phylogenetical clusters including 8 PHT1 homologs,1 PHT2 homolog, 2 PHT3 homologs, 5 PHT4 homologs and 4 PHO homologs. These genes were mapped on ten S. tuberosum chromosomes (chr1–9 and 12). Analysis of these genes led to identification of 20 conserved motifs and location prediction showed that PHT1 and PHT4 proteins have 12 transmembrane (TM) domains, PHT2 proteins have 13 TM domains, PHT3 proteins contain 4–5 TM domains, and PHO proteins contain 8–10 TM domains. Two tandem and 2 segmental duplications were further identified from these genes. RT-qPCR analysis showed that most of PHTs and PHOs displayed specific expression patterns with respect to phosphate status, fourteen transporters were up regulated in leaves and 11 transporters were up regulated in roots under phosphate deprivation. StPHT2;1 was found to be expressed both in potato leaf and root after phosphate starvation. Most cis -regulatory elements in StPHTs and StPHOs were found associated with light, defense, stress and hormones responsiveness, endosperm and meristem expression, as well as circadian control. A scan of the coding sequences of 20 StPHTs and StPHOs against published miRNAs in potato predicted a total of 2 potential miRNAs, mainly targeting members located on the same chromosome. Expression of StPHTs and StPHOs was selectively induced by biotic and abiotic stresses, and phytohormone treatments, among them, heat stress exclusively caused the up-regulation of 14 StPHT and StPHO genes, in particular PHO1 members. Most of StPHTs and StPHOs do not exhibit the tissue-specific expression. StPHT1;7, StPHT3;1 and StPHTO1;1 were highly expressed in the root of S. tuberosum , especially the gene StPHT3;1 are highly expressed in 14 tissues which covering the entire life cycle of potato. Our results provide comprehensive insights into the PHT and PHO family genes involved in the growth, development, and stress response of the potato. [ABSTRACT FROM AUTHOR]

Background: Trehalose-6-phosphate synthase (TPS) serves important functions in plant desiccation tolerance and response to environmental stimuli. At present, a comprehensive analysis, i.e. functional classification, molecular evolution, and expression patterns of this gene family are still lacking in Solanum tuberosum (potato). Results: In this study, a comprehensive analysis of the TPS gene family was conducted in potato. A total of eight putative potato TPS genes (StTPSs) were identified by searching the latest potato genome sequence. The amino acid identity among eight StTPSs varied from 59.91 to 89.54%. Analysis of dN/dS ratios suggested that regions in the TPP (trehalose-6-phosphate phosphatase) domains evolved faster than the TPS domains. Although the sequence of the eight StTPSs showed high similarity (2571-2796 bp), their gene length is highly differentiated (3189-8406 bp). Many of the regulatory elements possibly related to phytohormones, abiotic stress and development were identified in different TPS genes. Based on the phylogenetic tree constructed using TPS genes of potato, and four other Solanaceae plants, TPS genes could be categorized into 6 distinct groups. Analysis revealed that purifying selection most likely played a major role during the evolution of this family. Amino acid changes detected in specific branches of the phylogenetic tree suggests relaxed constraints might have contributed to functional divergence among groups. Moreover, StTPSs were found to exhibit tissue and treatment specific expression patterns upon analysis of transcriptome data, and performing qRT-PCR. Conclusions: This study provides a reference for genome-wide identification of the potato TPS gene family and sets a framework for further functional studies of this important gene family in development and stress response. [ABSTRACT FROM AUTHOR]

Cultivated potatoes (Solanum tuberosum L.), domesticated from wild Solanum species native to the Andes of southern Peru, possess a diverse gene pool representing more than 100 tuber-bearing relatives (Solanum section Petota). A diversity panel of wild species, landraces, and cultivars was sequenced to assess genetic variation within tuber-bearing Solanum and the impact of domestication on genome diversity and identify key loci selected for cultivation in North and South America. Sequence diversity of diploid and tetraploid S. tuberosum exceeded any crop resequencing study to date, in part due to expanded wild introgressions following polyploidy that captured alleles outside of their geographic origin. We identified 2,622 genes as under selection, with only 14-16% shared by North American and Andean cultivars, showing that a limited gene set drove early improvement of cultivated potato, while adaptation of upland (S. tuberosum group Andigena) and lowland (S. tuberosum groups Chilotanum and Tuberosum) populations targeted distinct loci. Signatures of selection were uncovered in genes controlling carbohydrate metabolism, glycoalkaloid biosynthesis, the shikimate pathway, the cell cycle, and circadian rhythm. Reduced sexual fertility that accompanied the shift to asexual reproduction in cultivars was reflected by signatures of selection in genes regulating pollen development/gametogenesis. Exploration of haplotype diversity at potato's maturity locus (StCDF1) revealed introgression of truncated alleles from wild species, particularly S. microdontum in long-day-adapted cultivars. This study uncovers a historic role of wild Solanum species in the diversification of long-day-adapted tetraploid potatoes, showing that extant natural populations represent an essential source of untapped adaptive potential. [ABSTRACT FROM AUTHOR]

Potato ( Solanum tuberosum L.) is a major crop worldwide and its productivity is severely reduced by high soil salinity. The aim of the current study was to gain insights into the response and regulation of the antioxidant system in the transgenic potato, overexpressing two genes viz. thermostable CuZn-superoxide dismutase ( PaSOD) gene from polyextremophile high-altitude plant Potentilla atrosanguinea and ascorbate peroxide ( RaAPX) gene from Rheum australe under salinity stress. Transgenic lines and wild type (WT) were assessed under 50, 100, and 150 mM concentrations of NaCl treatment at the anatomical, transcriptional and metabolic levels. Although superoxide dismutase (SOD) and ascorbate peroxide (APX) activities in transgenic lines were significantly higher than WT under salt stress, the secondary cell wall lignification was profoundly affected by the modulated antioxidant enzymes activities and the corresponding HO levels in transgenic. Expression profiles showed that the several genes and transcription factors directly involved in lignin biosynthesis were profoundly induced in transgenic lines as compared to WT. In addition, transgenic plants harbored increased starch accumulation, improved growth attributes, and reduced accumulation of reactive oxygen species than WT plants. Together, these results indicated that the expression of SOD and APX genes in transgenic potato may function as a positive set of physiological, anatomical, and molecular adjustments in the HO regulated lignin biosynthesis signaling pathway, enabling the plants to withstand severe saline conditions. [ABSTRACT FROM AUTHOR]

Key message: A tetraploid potato population was mapped for internal heat necrosis (IHN) using the Infinium 8303 potato SNP array, and QTL for IHN were identified on chromosomes 1, 5, 9 and 12 that explained 28.21% of the variation for incidence and 25.3% of the variation for severity. This research represents a significant step forward in our understanding of IHN, and sets the stage for future research focused on testing the utility of these markers in additional breeding populations. Abstract: Internal heat necrosis (IHN) is a significant non-pathogenic disorder of potato tubers and previous studies have identified AFLP markers linked to IHN susceptibility in the tetraploid, B2721 potato mapping population. B2721 consists of an IHN susceptible×resistant cross: Atlantic×B1829-5. We developed a next-generation SNP-based linkage map of this cross using the Infinium 8303 SNP array and conducted additional QTL analyses of IHN susceptibility in the B2721 population. Using SNP dosage sensitive markers, linkage maps for both parents were simultaneously analyzed. The linkage map contained 3427 SNPs and totaled 1397.68 cM. QTL were detected for IHN on chromosomes 1, 5, 9, and 12 using LOD permutation thresholds and colocation of high LOD scores across multiple years. Genetic effects were modeled for each putative QTL. Markers associated with a QTL were regressed in models of effects for IHN incidence and severity for all years. In the full model, the SNP markers were shown to have significant effects for IHN ( p < 0.0001), and explained 28.21% of the variation for incidence and 25.3% of the variation for severity. We were able to utilize SNP dosage information to identify and model the effects of putative QTL, and identify SNP loci associated with IHN resistance that need to be confirmed. This research represents a significant step forward in our understanding of IHN, and sets the stage for future research focused on testing the utility of these markers in additional breeding populations. [ABSTRACT FROM AUTHOR]

The expression of insecticidal genes must be induced at appropriate time and in sufficient amount to confer protection against targeted pests. However, the increased scientific reports of resistance development in insect pest against insecticidal delta-endotoxins, produced by Bacillus thuringiensis, provide impetus for the development of alternative insect management strategies. The present study was conducted to investigate the importance of targeted expression of a hybrid insecticidal gene (SN19) in potatoes. For this purpose, two plant expression vectors were constructed by cloning hybrid SN19 gene ( cry1Ba-domain I-III and cry1Ia-domain II) under the control of a wound-inducible promoter isolated from Asparagus officinalis (AoPR1) and CaMV 35S promoter, and were transferred to Agrobacterium tumefaciens strain EHA 105. Four potato genotypes (Marabel, Innovator, Tokat 10/1 and Tokat 6/24) were transformed with EHA 105 strain harboring pTF101.1 35S-SN19 and pTF101.1 AoPR1-SN19 constructs. Phosphinothricin (PPT) was used at concentration of 1 mg/l for selection of primary transformants. PCR results showed the presence of both introduced SN19 and bar genes in 43 plants out of total 154 putative transgenics. Expression of SN19 protein in primary transformants was confirmed by Western blot assays. The mechanical wounding of transgenic plants exhibited more accumulated levels of SN19 proteins during post wounding period. Leaf biotoxicity assays with Colorado potato beetle (Coleoptera) and tomato leafminer (Lepidoptera) exhibited 100% mortality of the pests in primary transformants. Based on our mortality results with both constructs, we concluded that the potato transgenic lines exhibited targeted expression of insecticidal gene under the control of AoPR1 promoter upon insect wounding with eliminated toxicity of Cry protein and hence can be further used effectively in potato breeding programme. [ABSTRACT FROM AUTHOR]

Genome editing (also known as targeted mutation) has promise for molecular breeding. Compared with the CRISPR/Cas9 system, the transcription activator-like effector nucleases (TALENs) have likely a lesser off-target rate in genome editing. Both a rapid test system for the functionality of designed TALENs and an effective delivery system for introducing the TALENs to plants are critical for successful target mutation. TALENs have usually been tested in protoplasts or introduced to plants with viral vectors. However, plant regeneration from protoplast culture can generate extensive somatic variation. Viral vectors are not always available, and plants edited by these vectors usually require virus elimination. Here, we used a non-viral, Agrobacterium-mediated transient expression approach, to serve both rapid test and effective delivery of TALENs into two vegetatively propagated potato cultivars, Solanum tuberosum Russet Burbank and Shepody. Two TALENs with different molecular weights (22 and 27 aa-repeat modules) were expressed to target two endogenous genes (starch branching enzyme and an acid invertase) by Agrobacterium-mediated infiltration (agroinfiltration) into leaves of potato plants. The infiltrated leaf DNA was analyzed using restriction site loss assay and subsequent DNA sequencing. Deep sequencing of these tetraploid cultivars was also conducted to determine the zygosity at the targeted chromosomal loci. TALENs, with different molecular weights, successfully agroinfiltrated and induced mutations at both targeted loci. [ABSTRACT FROM AUTHOR]

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DNA-based marker analysis of plant genebank material has become a useful tool in the evaluation of levels of genetic diversity and for the informed use and maintenance of germplasm. In this study, we quantify levels of amplified fragment length polymorphism (AFLP) in representative accessions of wild and cultivated potato species of differing geographic origin, ploidy, and breeding system. We generated 449 polymorphic AFLP fragments in 619 plants, representing multiple plants (16-23) from 17 accessions of 14 potato taxa as well as single plants sampled from available accessions (from 3 to 56) of the same 14 taxa. Intra-accession diversities were compared to those of a synthetic 'taxon-wide' population comprising a single individual from a variable number of available accessions of each sampled taxon. Results confirm the expected considerably lower levels of polymorphism within accessions of self-compatible as compared to self-incompatible taxa. We observed broadly similar levels of 'taxon-wide' polymorphism among self-compatible and self-incompatible species, with self-compatible taxa showing only slightly lower rates of polymorphism. The most diverse accessions were the two cultivated potato accessions examined, the least diverse being the Mexican allohexaploids Solanum demissum and S. iopetalum. Generally allopolyploid self-compatible accessions exhibited lower levels of diversity. Some purported self-incompatible accessions showed relatively low levels of marker diversity, similar to the more diverse self-compatible material surveyed. Our data indicate that for self-compatible species a single plant is highly representative of a genebank accession. The situation for self-incompatible taxa is less clear, and sampling strategies used will depend on the type of investigation. These results have important implications for those seeking novel trait variation (e.g., disease resistance) in gene banks as well as for the selection of individuals for genomics studies. We also show that AFLPs, despite having been largely replaced by other marker types, is highly suitable for the evaluation of within and between accession diversity in genebanks. [ABSTRACT FROM AUTHOR]

Streptomyces scabies is a common soil bacterium that causes scab symptoms in potatoes. Strong evidence indicates horizontal gene transfer (HGT) among bacteria has influenced the evolution of this plant pathogen and other Streptomyces spp. To extend the study of the HGT to the Streptomyces genus, we explored the effects of the inter-domain HGT in the S. scabies genome. We employed a semi-automatic pipeline based on BLASTp searches and phylogenetic reconstruction. The data show low impact of inter-domain HGT in the S. scabies genome; however, we found a putative plant pathogenesis related 1 (PR1) sequence in the genome of S. scabies and other species of the genus. It is possible that this gene could be used by S. scabies to out-compete other soil organisms. [ABSTRACT FROM AUTHOR]

Aiming at a better understanding of the physiological and biochemical background of nitrogen use efficiency, alterations in the shoot proteome under N-deficiency were investigated in two contrasting potato genotypes grown in vitro with 60 and 7.5 mM N, respectively. A gel based proteomic approach was applied to identify candidate proteins associated with genotype specific responses to N-deficiency. 21% of the detected proteins differed in abundance between the two genotypes. Between control and N-deficiency conditions 19.5% were differentially accumulated in the sensitive and 15% in the tolerant genotype. 93% of the highly N-deficiency responsive proteins were identified by MALDI TOF/TOF mass spectrometry. The major part was associated with photosynthesis, carbohydrate metabolism, stress response and regulation. Differential accumulation of enzymes involved in the Calvin cycle and glycolysis suggest activation of alternative carbohydrate pathways. In the tolerant genotype, increased abundance under N-deficiency was also found for enzymes involved in chlorophyll synthesis and stability of enzymes, which increase photosynthetic carbon fixation efficiency. Out of a total of 106 differentially abundant proteins, only eight were detected in both genotypes. Our findings suggest that mutually responsive proteins reflect universal stress responses while adaptation to N-deficiency in metabolic pathways is more genotype specific. Significance Nitrogen losses from arable farm land considerably contribute to environmental pollution. In potato, this is a special problem due cultivation on light soils, irrigation and the shallow root system. Therefore, breeding of cultivars with improved nitrogen use efficiency and stable yields under reduced N fertilization is an important issue. Knowledge of genotype dependent adaptation to N-deficiency at the proteome level can help to understand regulation of N efficiency and development of N-efficient cultivars. [ABSTRACT FROM AUTHOR]

‘Candidatus Liberibacter solanacearum’ (Lso), an uncultivable phloem-limited phytopathogenic bacteria, is known to be associated with Zebra Chip disease (ZC), which represents a major threat to potato production in the US and elsewhere. This pathogen is transmitted by the phloem-feeding potato psyllid, Bactericera cockerelli Sulc (Hem. Triozidae). Currently, there are no reports of resistance to ZC in cultivated potatoes. This greenhouse study was conducted to evaluate Lso transmission success and the susceptibility of 11 potato breeding clones, representing diverge genetic background, to ZC, in relation to a susceptible commercial cultivar, Russet Burbank. Individual plants were exposed to two Lso-positive potato psyllids for 48 hours. The percentage of successful Lso transmission varied across the evaluated genotypes ranging between 7 and 57%. Freshly-cut and fried tubers showed significant variation in ZC symptom severity among the breeding clones, with several genotypes expressing relative tolerance when compared to Russet Burbank. None of the evaluated clones showed statistically lower Lso titers than Russet Burbank with the exception of one genotype in the second year of the study. However, the presence of a non-significant relationship between average symptom severity and Lso titer indicated variations in phenotypic responses (i.e. tolerance) to Lso existed among evaluated breeding lines. Breeding clones A07781-3LB, A07781-4LB and A07781-10LB had relatively lower Lso titer (low susceptibility) and tuber symptom expression (high tolerance) among the tested genotypes. As these three clones represent full siblings, the observed effects could be indicative of the presence of a genetic basis for resistance/tolerance to ZC. Findings provide a better understanding of resistance/tolerance to ZC, and contribute to continued efforts in breeding for resistance to this disease. [ABSTRACT FROM AUTHOR]

Plant root secreted compounds alter the gene expression of associated microorganisms by acting as signal molecules that either stimulate or repel the interaction with beneficial or harmful species, respectively. However, it is still unclear whether two distinct groups of beneficial bacteria, non-plant-associated (soil) strains and plantassociated (endophytic) strains, respond uniformly or variably to the exposure with root exudates. Therefore, Bacillus mycoides, a potential biocontrol agent and plant growth-promoting bacterium, was isolated from the endosphere of potatoes and from soil of the same geographical region. Confocal fluorescence microscopy of plants inoculated with GFP-tagged B. mycoides strains showed that the endosphere isolate EC18 had a stronger plant colonization ability and competed more successfully for the colonization sites than the soil isolate SB8. To dissect these phenotypic differences, the genomes of the two strains were sequenced and the transcriptome response to potato root exudates was compared. The global transcriptome profiles evidenced that the endophytic isolate responded more pronounced than the soil-derived isolate and a higher number of significant differentially expressed genes were detected. Both isolates responded with the alteration of expression of an overlapping set of genes, which had previously been reported to be involved in plant-microbe interactions; including organic substance metabolism, oxidative reduction, and transmembrane transport. Notably, several genes were specifically upregulated in the endosphere isolate EC18, while being oppositely downregulated in the soil isolate SB8. These genes mainly encoded membrane proteins, transcriptional regulators or were involved in amino acid metabolism and biosynthesis. By contrast, several genes upregulated in the soil isolate SB8 and downregulated in the endosphere isolate EC18 were related to sugar transport, which might coincide with the different nutrient availability in the two environments. Altogether, the presented transcriptome profiles provide highly improved insights into the life strategies of plant-associated endophytes and soil isolates of B. mycoides. [ABSTRACT FROM AUTHOR]

Key message: Transient 5-azacytidine treatment of leaf explants from potato plants with transcriptionally silenced transgenes allows de novo regeneration of plants with restored transgene expression at the whole plant level. Abstract: Transgenes introduced into plant genomes frequently become silenced either at the transcriptional or the posttranscriptional level. Transcriptional silencing is usually associated with DNA methylation in the promoter region. Treatments with inhibitors of maintenance DNA methylation were previously shown to allow reactivation of transcriptionally silenced transgenes in single cells or tissues, but not at the whole plant level. Here we analyzed the effect of DNA methylation inhibitor 5-azacytidine (AzaC) on the expression of two silenced reporter genes encoding green fluorescent protein (GFP) and neomycin phosphotransferase (NPTII) in potato plants. Whereas no obvious reactivation was observed in AzaC-treated stem cuttings, transient treatment of leaf segments with 10 μM AzaC and subsequent de novo regeneration of shoots on the selective medium with kanamycin resulted in the production of whole plants with clearly reactivated expression of previously silenced transgenes. Reactivation of nptII expression was accompanied by a decrease in cytosine methylation in the promoter region of the gene. Using the plants with reactivated GFP expression, we found that re-silencing of this transgene can be accidentally triggered by de novo regeneration. Thus, testing the incidence of transgene silencing during de novo regeneration could be a suitable procedure for negative selection of transgenic lines (insertion events) which have an inclination to be silenced. Based on our analysis of non-specific inhibitory effects of AzaC on growth of potato shoots in vitro, we estimated that AzaC half-life in the culture media is approximately 2 days. [ABSTRACT FROM AUTHOR]

Nuclear factor Y (NF-Y) is one of the most ubiquitous transcription factors (TFs), comprising NF-YA, NF-YB and NF-YC subunits, and has been identified and reported in various aspects of development for plants and animals. In this work, StNF-YB3.1 , a putative potato NF-YB subunit encoding gene, was isolated from Solanum tuberosum by rapid amplification of cDNA ends (RACE). Overexpression of StNF-YB3.1 in potato (cv. Atlantic) resulted in accelerated onset of flowering, and significant increase in leaf chlorophyll content in field trials. However, transgenic potato plants overexpressing StNF-YB3.1 (OEYB3.1) showed significant decreases in photosynthetic rate and stomatal conductance both at tuber initiation and bulking stages. OEYB3.1 lines were associated with significantly fewer tuber numbers and yield reduction. Guard cell size and stomatal density were not changed in OEYB3.1 plants, whereas ABA-mediated stomatal closure was accelerated compared to that of wild type plants because of the up-regulation of genes for ABA signaling, such as StCPK10-like , StSnRK2.6/OST1-like , StSnRK2.7-like and StSLAC1-like . We speculate that the acceleration of stomatal closure was a possible reason for the significantly decreased stomatal conductance and photosynthetic rate. [ABSTRACT FROM AUTHOR]

Tuber formation of potato is a complex biological process and is regulated by many factors including phytochrome, hormones, transcription factors, RNAs, microRNAs, etc. In our previous study, CYP707A1, an ABA 8′-hydroxylase gene, was down-regulated in the StCOL antisense transgenic potato stolons, but its relation with potato tuberization has not been studied yet. In this study, to investigate the role of this gene in potato tuberization, we cloned a CYP707A1 gene from potato cultivar Désirée and constructed the StCYP707A1 over-expression and anti-sense potato lines by Agrobacterium-mediated transformation. In total, four over-expression transgenic lines and nine anti-sense transgenic lines were confirmed by PCR analysis. Semi-quantitative RT-PCR and qRT-PCR analysis showed that the level of StCYP707A1 transcripts was significantly higher in the over-expression lines (A7 and A9) and lower in the anti-sense lines (F2 and F8), compared to the wild-type control. We further analyzed tuber formation in the transgenic lines and the wild-type control. The result showed that tuber yield per plant and average tuber weight were decreased in A7 and A9 and increased in F2. We also measured the content of ABA and GA in transgenic lines. ABA level was reduced in A7 and A9 and increased in F2. Contrariwise, the concentration of GA was higher in A7 and A9, and lower in F2 than in wild-type control. These results indicate that StCYP707A1 negatively affects potato tuberization through ABA regulation on gibberellic acid concentration. [ABSTRACT FROM AUTHOR]

This study presents the characterization of the plasma membrane (PM) H+-ATPases in potato, focusing on their role in stolon and tuber development. Seven PM H+-ATPase genes were identified in the Solanum tuberosum genome, designated PHA1-PHA7. PHA genes show distinct expression patterns in different plant tissues and under different stress treatments. Application of PM H+-ATPase inhibitors arrests stolon growth, promotes tuber induction, and reduces tuber size, indicating that PM H+-ATPases are involved in tuberization, acting at different stages of the process. Transgenic potato plants overexpressing PHA1 were generated (PHA1-OE). At early developmental stages, PHA1-OE stolons elongate faster and show longer epidermal cells than wild-type stolons; this accelerated growth is accompanied by higher cell wall invertase activity, lower starch content, and higher expression of the sucrose-H+ symporter gene StSUT1. PHA1-OE stolons display an increased branching phenotype and develop larger tubers. PHA1-OE plants are taller and also present a highly branched phenotype. These results reveal a prominent role for PHA1 in plant growth and development. Regarding tuberization, PHA1 promotes stolon elongation at early stages, and tuber growth later on. PHA1 is involved in the sucrose-starch metabolism in stolons, possibly providing the driving force for sugar transporters to maintain the apoplastic sucrose transport during elongation. [ABSTRACT FROM AUTHOR]