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We have extended previously published sets of simple sequence repeat markers for Synchytrium endobioticum, selected to be polymorphic for the German-standard isolates of pathotypes P1, P2, P6, P8, and P18. These markers also complement the extensive published information on DNA polymorphisms for the mitogenomes of Synchytrium endobioticum. This extended set of 35 markers representing 73 alleles differentiated 51 isolates from Europe and North America into three large, well-separated clusters and subclusters using dendrogram analysis, principal coordinates analysis (PCoA), and population substructure analysis using STRUCTURE 2.3.4 software. This suggests a limited number of introgressions of the wart disease pathogen into current potato growing areas, followed by recombination and admixture of populations through human activities. The new markers extend the published marker sets and are useful tools for future analyses of population structure and dynamics in Synchytrium endobioticum, which are necessary to understand the biology of the interaction between the pathogen and its potato host and to develop future control strategies. [ABSTRACT FROM AUTHOR]

Phosphorous starvation in plants has been reported to have contrasting effects on the interaction with pathogens in different plant pathogen systems and plant species. Both increases and decreases in susceptibility have been observed in numerous reports. Here, we analysed black spot infection and the leaf expression of two plant phosphate transporters and one defence marker gene in roses after phosphorous starvation. We varied three factors: phosphate starvation versus full supply of phosphorous, black spot infection vs. mock inoculation, and different susceptible and resistant progeny of a biparental rose population. Black spot susceptibility or resistance was not significantly changed upon phosphate starvation in either compatible or incompatible interactions. The expression of phosphate transporters was strongly induced upon starvation, but in some genotypes, expression was altered by black spot interaction as well. The marker for pathogenic interactions was exclusively induced by interaction with black spot, but the expression was altered by a combination of phosphate starvation and interaction with the fungus in some genotypes. In summary, phosphate starvation has clear effects on the gene expression of phosphate transporters in rose leaves, and the interaction with a hemibiotrophic leaf pathogen is strongly genotype dependent. [ABSTRACT FROM AUTHOR]

Abstract Background The resistance of plants to pathogens relies on two lines of defense: a basal defense response and a pathogen-specific system, in which resistance (R) genes induce defense reactions after detection of pathogen-associated molecular patterns (PAMPS). In the specific system, a so-called arms race has developed in which the emergence of new races of a pathogen leads to the diversification of plant resistance genes to counteract the pathogens’ effect. The mechanism of resistance gene diversification has been elucidated well for short-lived annual species, but data are mostly lacking for long-lived perennial and clonally propagated plants, such as roses. We analyzed the rose black spot resistance gene, Rdr1, in five members of the Rosaceae: Rosa multiflora, Rosa rugosa, Fragaria vesca (strawberry), Malus x domestica (apple) and Prunus persica (peach), and we present the deduced possible mechanism of R-gene diversification. Results We sequenced a 340.4-kb region from R. rugosa orthologous to the Rdr1 locus in R. multiflora. Apart from some deletions and rearrangements, the two loci display a high degree of synteny. Additionally, less pronounced synteny is found with an orthologous locus in strawberry but is absent in peach and apple, where genes from the Rdr1 locus are distributed on two different chromosomes. An analysis of 20 TIR-NBS-LRR (TNL) genes obtained from R. rugosa and R. multiflora revealed illegitimate recombination, gene conversion, unequal crossing over, indels, point mutations and transposable elements as mechanisms of diversification. A phylogenetic analysis of 53 complete TNL genes from the five Rosaceae species revealed that with the exception of some genes from apple and peach, most of the genes occur in species-specific clusters, indicating that recent TNL gene diversification began prior to the split of Rosa from Fragaria in the Rosoideae and peach from apple in the Spiraeoideae and continued after the split in individual species. Sequence similarity of up to 99% is obtained between two R. multiflora TNL paralogs, indicating a very recent duplication. Conclusions The mechanisms by which TNL genes from perennial Rosaceae diversify are mainly similar to those from annual plant species. However, most TNL genes appear to be of recent origin, likely due to recent duplications, supporting the hypothesis that TNL genes in woody perennials are generally younger than those from annuals. This recent origin might facilitate the development of new resistance specificities, compensating for longer generation times in woody perennials.

The Rdr1 gene confers resistance to black spot in roses and belongs to a large TNL gene family, which is organized in two major clusters at the distal end of chromosome 1. We used the recently available chromosome scale assemblies for the R. chinensis 'Old Blush' genome, re-sequencing data for nine rose species and genome data for Fragaria, Rubus, Malus and Prunus to identify Rdr1 homologs from different taxa within Rosaceae. Members of the Rdr1 gene family are organized into two major clusters in R. chinensis and at a syntenic location in the Fragaria genome. Phylogenetic analysis indicates that the two clusters existed prior to the split of Rosa and Fragaria and that one cluster has a more recent origin than the other. Genes belonging to cluster 2, such as the functional Rdr1 gene muRdr1A, were subject to a faster evolution than genes from cluster 1. As no Rdr1 homologs were found in syntenic positions for Prunus persica, Malus x domestica and Rubus occidentalis, a translocation of the Rdr1 clusters to the current positions probably happened after the Rubeae split from other groups within the Rosoideae approximately 70–80 million years ago during the Cretaceous period. [ABSTRACT FROM AUTHOR]

We studied the petal number trait in a population obtained after self-pollination of R. x damascena f. trigintipetala following analysis of molecular markers which have previously been mapped near the major dominant locus Blfo/d6 determining this trait in other rose species including R. multiflora and R. hybrida. The results showed that the same genetic mechanism, which determines the petal number trait in R. multiflora and R. hybrida also controls the trait in R. x damascena f. trigintipetala and is related to the dominant effect of a single copy allele in the tetraploid genome of this species. We also analyzed the expression of several flower homeotic genes including R. x damascena APETALA1/FUL-like (paleo AP1 type), R. x damascena euAPETALA 3 (euAP3 line) and R. x damascena AGAMOUS in early stage flower buds corresponding to plants with double and simple flowers. The obtained results showed that only R. x damascena AGAMOUS was differentially expressed between the samples of double and simple flowers, its relative expression being upregulated 3.5-fold in simple flowers. We further cloned and sequenced the four genomic clones of R. x damascena AGAMOUS and studied the potential additive effect of this gene by analysing the segregation of its four alleles in the population of self-pollinated R. x damascena. Analysis of variance of the data for petal number and allele segregation did not show a statistically significant effect of any allele configuration of the AGAMOUS gene on the petal number trait in R. x damascena f. trigintipetala. [ABSTRACT FROM AUTHOR]

African nightshade is an indigenous leafy vegetable in Eastern and Western Africa that is valued for its high nutrient content and potential to generate income. Efforts to improve this crop have been hampered by limited information available on intra- and interspecific genetic diversity. Here, we present data on the development and application of polymorphic simple sequence repeat (SSR) markers in a broad collection of entries (gene bank accessions, landraces from farmers, breeding lines and commercial cultivars) from two species of African nightshades, Solanum scabrum (Mill.) and S. villosum (Mill.). Next generation sequencing information was used to develop 16 polymorphic SSR markers that revealed genetic diversity within and between the 54 entries collected from various geographic locations in Sub-Saharan Africa. High levels of polymorphism were detected within entries, indicating gene flow between entries and/or a low intensity of selection during development of breeding lines and improved cultivars. As a consequence, they have retained sufficient variation for further selection within the improved germplasm. The two species were clearly distinct, and S. scabrum was found to be less diverse compared to S. villosum . Furthermore, some entries were identified as probably misclassified based on morphological markers as they can clearly be placed within the gene pool of either S. scabrum or S. villosum using the SSR data. In addition to the diversity studies, we determined the genome size and ploidy of a selection of 29 entries helpful for the design of future NGS-based marker projects. All entries of S. scabrum were found to be hexaploid, whereas entries of S. villosum were tetraploid with a haploid genome size of all samples slightly varying by approximately 1 pg. Finally, we also determined pollen viability parameters, which differed significantly between the two species but did not correlate to any of the other parameters measured. [ABSTRACT FROM AUTHOR]

Haploid potato populations derived from a single tetraploid donor constitute an efficient strategy to analyze markers segregating from a single donor genotype. Analysis of marker segregation in populations derived from crosses between polysomic tetraploids is complicated by a maximum of eight segregating alleles, multiple dosages of the markers and problems related to linkage analysis of marker segregation in repulsion. Here, we present data on two monoparental haploid populations generated by prickle pollination of two tetraploid cultivars with Solanum phureja and genotyped with the 12.8 k SolCAP single nucleotide polymorphism (SNP) array. We show that in a population of monoparental haploids, the number of biallelic SNP markers segregating in linkage to loci from the tetraploid donor genotype is much larger than in putative crosses of this genotype to a diverse selection of 125 tetraploid cultivars. Although this strategy is more laborious than conventional breeding, the generation of haploid progeny for efficient marker analysis is straightforward if morphological markers and flow cytometry are utilized to select true haploid progeny. The level of introgressed fragments from S. phureja, the haploid inducer, is very low, supporting its suitability for genetic analysis. Mapping with single-dose markers allowed the analysis of quantitative trait loci (QTL) for four phenotypic traits. [ABSTRACT FROM AUTHOR]

Summary: Black spot disease, which is caused by the ascomycete Diplocarpon rosae, is the most severe disease in field‐grown roses in temperate regions and has been distributed worldwide, probably together with commercial cultivars. Here, we present data indicating that muRdr1A is the active Rdr1 gene, a single‐dominant TIR‐NBS‐LRR (Toll/interleukin‐1 receptor‐nucleotide binding site‐leucine rich repeat) (TNL)‐type resistance gene against black spot disease, which acts against a broad range of pathogenic isolates independent of the genetic background of the host genotype. Molecular analyses revealed that, compared with the original donor genotype, the multiple integrations that are found in the primary transgenic clone segregate into different integration patterns in its sexual progeny and do not show any sign of overexpression. Rdr1 provides resistance to 13 different single‐spore isolates belonging to six different races and broad field mixtures of conidia; thus far, Rdr1 is only overcome by two races. The expression of muRdr1A, the active Rdr1 gene, leads to interaction patterns that are identical in the transgenic clones and the non‐transgenic original donor genotype. This finding indicates that the interacting avirulence (Avr) factor on the pathogen side must be widespread among the pathogen populations and may have a central function in the rose–black spot interaction. Therefore, the Rdr1 gene, pyramided with only a few other R genes by sexual crosses, might be useful for breeding roses that are resistant to black spot because the spread of new pathogenic races of the fungus appears to be slow. [ABSTRACT FROM AUTHOR]

African nightshades are becoming more important as leafy vegetables in sub-Saharan Africa. Previously considered as food for the poor, their cultivation is now being promoted, and some cultivars are commercialised; however, most farmers use self-produced seeds, leading to low and varying yields. Improvement through conventional breeding depends on the available genetic diversity, the possible breeding systems, and the nutritional value of the accessions. Therefore, we review the information on these topics with the following main outcomes: the most commonly discussed species,S. nigrum, S. scabrum, S. villosum, andS. americanum, could be differentiated using molecular markers, but further sub-clustering was rarely possible, and statistical support often missing.S. nigrumandS. scabrumseem to be most closely related to each other. The mainly self-pollinating African nightshades form a polyploidy series with diploid (2n = 2x = 24) to hexaploid taxa. Interploidy hybridisations between diploids and tetraploids are possible, whereas the hexaploidS. nigrumandS. scabrumcould not be crossed to genotypes of lower ploidies. Solanine, solamargine, solasonine, and chaconine are the major steroidal alkaloid glucosides in African nightshades. Amounts are age and environment dependant. Mineral and vitamin contents in leaves are at least as high as inBrassica oleraceaorSpinacia oleracea, underlining their relevance as local vegetables. [ABSTRACT FROM AUTHOR]

Cowpea ( Vigna unguiculata (L.) Walp.) is widely grown by subsistence farmers in West and East Africa where its grain and leaves are sources of highly valuable food, due to their high contents of proteins, minerals and vitamins. Therefore, cowpea could play a significant role in mitigating malnutrition such as micronutrient deficiencies. The objectives of this study were to evaluate the performance in agronomic traits of cowpea entries (released and farmers’ cultivars, and gene bank accessions) from different sources, and to assess the extent of genetic diversity in this material to provide basic information for its use in breeding programs. A total of 15 entries were evaluated. All entries were morphologically uniform, except the farmers’ local cultivars. Fresh leaf yield varied from 34.6 to 52.8 g per plant and days to 50% flowering from 64 to 82 days. Hundred seed weight ranged from 7.67 to 15.12 g. On average, the number of pods ranged from 4.8 to 15.6 pods per plant. No correlation between fresh leaf yield and other traits was detected, whereas the number of pods per plant and the hundred seed weight were negatively correlated. Genetic diversity was assessed on five genotypes per entry using 544 Amplified Fragment Length Polymorphism (AFLP) and 18 microsatellite markers. Genetic distances calculated using the Jaccard algorithm ranged from 0.002 to 0.193 among genotypes of the same entry and from 0.098 to 0.301 for genotypes from different entries. A principle coordinate analysis separated four entries from the rest. Although the consensus tree based on Neighbor Joining trees was unable to resolve the whole cluster, an assignment of most of the entries into entry-specific clades was possible. [ABSTRACT FROM AUTHOR]

Cleome gynandra (L.) Briq. is an African leafy vegetable with a potential to improve food security and micronutrient deficiencies. Cytological traits, breeding biology and genetic diversity of 30 selected entries of C. gynandra from six African countries were investigated. The entries consisted of advanced lines, gene bank accessions and farmers' cultivars. Our study revealed chromosome numbers of 2 n = 34 in root tip metaphase cells from one entry. The 30 entries were found to be diploid with genome sizes ranging from 2.31 to 2.45 pg/2C. Hand pollination experiments were carried out to assess self-incompatibility within the entries and revealed that they are self- and cross-compatible. For genetic diversity studies within and among the entries, the pooled data of 499 polymorphic bands from 11 amplified fragment length polymorphism primer combinations and nine simple sequence repeat markers were used. The genetic distance among the entries ranged from 0.13 to 0.77. In a principal coordinate analysis, the farmers' cultivars formed a cluster separate from the advanced lines and the gene bank entries, and the latter were not well resolved. [ABSTRACT FROM AUTHOR]

Petal color is one of the key characteristics determining the attractiveness and therefore the commercial value of an ornamental crop. Here, we present the first genome-wide association study for the important ornamental crop rose, focusing on the anthocyanin and carotenoid contents in petals of 96 diverse tetraploid garden rose genotypes. Cultivated roses display a vast phenotypic and genetic diversity and are therefore ideal targets for association genetics. For marker analysis, we used a recently designed Axiom SNP chip comprising 68,000 SNPs with additionally 281 SSRs, 400 AFLPs and 246 markers from candidate genes. An analysis of the structure of the rose population revealed three subpopulations with most of the genetic variation between individual genotypes rather than between clusters and with a high average proportion of heterozygous loci. The mapping of markers significantly associated with anthocyanin and carotenoid content to the related Fragaria and Prunus genomes revealed clusters of associated markers indicating five genomic regions associated with the total anthocyanin content and two large clusters associated with the carotenoid content. Among the marker clusters associated with the phenotypes, we found several candidate genes with known functions in either the anthocyanin or the carotenoid biosynthesis pathways. Among others, we identified a glutathione-S-transferase, 4CL, an auxin response factor and F3'H as candidate genes affecting anthocyanin concentration, and CCD4 and Zeaxanthine epoxidase as candidates affecting the concentration of carotenoids. These markers are starting points for future validation experiments in independent populations as well as for functional genomic studies to identify the causal factors for the observed color phenotypes. Furthermore, validated markers may be interesting tools for marker-assisted selection in commercial breeding programmes in that they provide the tools to identify superior parental combinations that combine several associated markers in higher dosages. [ABSTRACT FROM AUTHOR]

Background: Interspecific hybridization is a useful tool in ornamental breeding to increase genetic variability and introduce new valuable traits into existing cultivars. The successful formation of interspecific hybrids is frequently limited by the presence of pre- and post-fertilization barriers. In the present study, we investigated the nature of hybridization barriers occurring in crosses between Kalanchoë species and evaluated possibilities of obtaining interspecific hybrids. Results: The qualitative and quantitative analyses of pollen tube growth in situ were performed following intra- and interspecific pollinations. They revealed occurrence of pre-fertilization barriers associated with inhibition of pollen germination on the stigma and abnormal growth of pollen tubes. Unilateral incongruity related to differences in pistil length was also observed. The pollen quality was identified as a strong factor influencing the number of pollen tubes germinating in the stigma. In relation to post-fertilization barriers, endosperm degeneration was a probable barrier hampering production of interspecific hybrids. Moreover, our results demonstrate the relation of genetic distance estimated by AFLP marker analysis of hybridization partners with cross-compatibility of Kalanchoë species. At the same time, differences in ploidy did not influence the success of interspecific crosses. Conclusions: Our study presents the first comprehensive analysis of hybridization barriers occurring within Kalanchoë genus. Reproductive barriers were detected on both, pre- and post-fertilization levels. This new knowledge will contribute to further understanding of reproductive isolation of Kalanchoë species and facilitate breeding of new cultivars. For the first time, interspecific hybrids between K. nyikae as maternal plant and K. blossfeldiana as well as K. blossfeldiana and K. marnieriana were generated. [ABSTRACT FROM AUTHOR]

Polyploidy is a widespread phenomenon among higher plants and a major factor shaping the structure and evolution of plant genomes. The important ornamental chrysanthemum (Chrysanthemum indicum hybrid) possesses a hexaploid genome with 54 chromosomes and was classified based on its evolutionary origin and cytological methods as an allopolyploid. However, it is questionable whether cytological methods are sufficient to determine the type of ploidy, and there are more informative methods available based on molecular marker analyses. Therefore, we collected segregation data for 406 dominant molecular marker alleles [327 amplified fragment length polymorphism (AFLPs), 65 single-strand conformation polymorphism (SSCPs) and 14 microsatellites (EST-SSRs)] in a biparental F1 population of 160 individuals.We analyzed these data for the characteristics that differ between allopolyploids and autopolyploids, including the segregation ratio of each marker, the ratio of single-dose (SD) to multi-dose (MD) markers, the ratio of SD markers in coupling to those in repulsion and the banding patterns of the SSRs. Whereas the analysis of the segregation ratio of each polymorphic marker indicated disomic (13 markers) as well as hexasomic (eight markers) inheritance, the ratio of SD markers in coupling to those in repulsion was 1:0, which is characteristic of autopolyploids. The observed ratio of SD to MD markers was 0.67:0.33 which is significantly different to the expected segregation for auto- and allohexaploids. Furthermore, the three EST-SSR alleles were inherited in all possible combinations and were not independent of each other, as expected for fixed heterozygosity in allopolyploids. Combining our results with published cytological data indicates that cultivated chrysanthemums should be classified as segmental allohexaploids. [ABSTRACT FROM AUTHOR]

The detection of QTL by association genetics depends on the genetic architecture of the trait under study, the size and structure of the investigated population and the availability of phenotypic and marker data of sufficient quality and quantity. In roses, we previously demonstrated that major QTL could already be detected in small association panels. In this study, we analyzed petal number, petal size and fragrance in a small panel of 95 mostly tetraploid garden rose genotypes. After genotyping the panel with the 68 K Axiom WagRhSNP chip we detected major QTL for all three traits. Each trait was significantly influenced by several genomic regions. Some of the QTL span genomic regions that comprise several candidate genes. Selected markers from some of these regions were converted into KASP markers and were validated in independent populations of up to 282 garden rose genotypes. These markers demonstrate the robustness of the detected effects independent of the set of genotypes analyzed. Furthermore, the markers can serve as tools for marker-assisted breeding in garden roses. Over an extended timeframe, they may be used as a starting point for the isolation of the genes underlying the QTL. [ABSTRACT FROM AUTHOR]

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Highlights • Out of 25 putative aquaporin genes in the sweet cherry genome 16 are expressed in fruit. • Most AQP genes are down-regulated at maturity, exceptions are PaPIP1;2 and PaPIP1;4. • Expression of AQP genes in the flesh is generally higher than that in the skin. • Water transport is blocked by the AQP inhibitors HgCl 2 , AgNO 3 , H 2 O 2 and Na acetate. Abstract Aquaporins (AQPs) function as gated membrane channels for water. They may have a role in water transport in sweet cherries (Prunus avium L.) where rain-induced cracking and the associated water uptake, severely limit production. The objectives were (1) to identify potential AQP genes in sweet cherry, (2) to establish their expression patterns during development and (3) to investigate their roles in the transmembrane movement of water at maturity. Based on an expression database of developing fruit and the recently published sweet cherry genome, 25 putative full length AQP genes were identified and 16 of them were expressed in the fruit. These 16 putative genes included seven PIPs, three TIPs, three NIPs and three SIPs. Most of them had six transmembrane domains and asparagine-proline-alanine motifs characteristic for AQPs. Expression differed among AQPs, tissues and developmental stages. The most highly expressed AQPs were PaPIP1;2 , PaPIP1;4 and PaTIP1;1. Expression in the flesh generally exceeded that in the skin. Furthermore, expression decreased towards maturity. Incubating flesh discs in hypertonic sucrose solutions resulted in water efflux from the disc. HgCl 2 (1 mM), AgNO 3 (1 mM), H 2 O 2 (100 mM) or Na acetate (100 mM) all decreased the rate of efflux, presumably through an inhibition of AQPs. Lower inhibitor concentrations had no significant effects. The results indicate: (1) that AQPs are expressed in sweet cherry fruits, (2) that expression is generally higher in the flesh than the skin and decreases towards maturity and (3) that AQPs are involved in water transport across plasma membranes. [ABSTRACT FROM AUTHOR]

Synchytrium endobioticum is an obligate biotrophic fungus that causes wart diseases in potato. Like other species of the class Chytridiomycetes, it does not form mycelia and its zoospores are small, approximately 3 μm in diameter, which complicates the detection of early stages of infection. Furthermore, potato wart disease is difficult to control because belowground organs are infected and resting spores of the fungus are extremely durable. Thus, S. endobioticum is classified as a quarantine organism. More than 40 S. endobioticum pathotypes have been reported, of which pathotypes 1(D1), 2(G1), 6(O1), 8(F1), and 18(T1) are the most important in Germany. No molecular methods for the differentiation of pathotypes are available to date. In this work, we sequenced both genomic DNA and cDNA of the German pathotype 18(T1) from infected potato tissue and generated 5,422 expressed sequence tags (EST) and 423 genomic contigs. Comparative sequencing of 33 genes, single-stranded confirmation polymorphism (SSCP) analysis with polymerase chain reaction fragments of 27 additional genes, as well as the analysis of 41 simple sequence repeat (SSR) loci revealed extremely low levels of variation among five German pathotypes. From these markers, one sequence-characterized amplified region marker and five SSR markers revealed polymorphisms among the German pathotypes and an extended set of 11 additional European isolates. Pathotypes 8(F1) and 18(T1) displayed discrete polymorphisms which allow their differentiation from other pathotypes. Overall, using the information of the six markers, the 16 isolates could be differentiated into three distinct genotype groups. In addition to the presented markers, the new collection of EST from genus Synchytrium might serve in the future for molecular taxonomic studies as well as for analyses of the host-pathogen interactions in this difficult pathosystem. [ABSTRACT FROM AUTHOR]