39 results on '"Veilleux RE"'
Search Results
2. Characterization of the F Locus Responsible for Floral Anthocyanin Production in Potato.
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Laimbeer FPE, Bargmann BOR, Holt SH, Pratt T, Peterson B, Doulis AG, Buell CR, and Veilleux RE
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- Flowers genetics, Flowers metabolism, Gene Expression Regulation, Plant, Plant Proteins genetics, Plant Proteins metabolism, Plants, Genetically Modified, Anthocyanins, Solanum tuberosum genetics
- Abstract
Anthocyanins are pigmented secondary metabolites produced via the flavonoid biosynthetic pathway and play important roles in plant stress responses, pollinator attraction, and consumer preference. Using RNA-sequencing analysis of a cross between diploid potato ( Solanum tuberosum L.) lines segregating for flower color, we identified a homolog of the ANTHOCYANIN 2 ( AN2 ) gene family that encodes a MYB transcription factor, herein termed StFlAN2 , as the regulator of anthocyanin production in potato corollas. Transgenic introduction of StFlAN2 in white-flowered homozygous doubled-monoploid plants resulted in a recovery of purple flowers. RNA-sequencing revealed the specific anthocyanin biosynthetic genes activated by StFlAN2 as well as expression differences in genes within pathways involved in fruit ripening, senescence, and primary metabolism. Closer examination of the locus using genomic sequence analysis revealed a duplication in the StFlAN2 locus closely associated with gene expression that is likely attributable to nearby genetic elements. Taken together, this research provides insight into the regulation of anthocyanin biosynthesis in potato while also highlighting how the dynamic nature of the StFlAN2 locus may affect expression., (Copyright © 2020 Laimbeer et al.)
- Published
- 2020
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3. Genome-wide Inference of Somatic Translocation Events During Potato Dihaploid Production.
- Author
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Pham GM, Braz GT, Conway M, Crisovan E, Hamilton JP, Laimbeer FPE, Manrique-Carpintero N, Newton L, Douches DS, Jiang J, Veilleux RE, and Buell CR
- Subjects
- Tetraploidy, Chromosomes, Plant, Diploidy, Plant Breeding, Solanum tuberosum genetics, Translocation, Genetic
- Abstract
Potato ( L.) breeders often use dihaploids, which are 2× progeny derived from 4× autotetraploid parents. Dihaploids can be used in diploid crosses to introduce new genetic material into breeding germplasm that can be integrated into tetraploid breeding through the use of unreduced gametes in 4× by 2× crosses. Dihaploid potatoes are usually produced via pollination by haploid inducer lines known as in vitro pollinators (IVP). In vitro pollinator chromosomes are selectively degraded from initially full hybrid embryos, resulting in 2× seed. During this process, somatic translocation of IVP DNA may occur. In this study, a genome-wide approach was used to identify such events and other chromosome-scale abnormalities in a population of 95 dihaploids derived from a cross between potato cultivar Superior and the haploid inducing line IVP101. Most Superior dihaploids showed translocation rates of <1% at 16,947,718 assayable sites, yet two dihaploids showed translocation rates of 1.86 and 1.60%. Allelic ratios at translocation sites suggested that most translocations occurred in individual cell lineages and were thus not present in all cells of the adult plants. Translocations were enriched in sites associated with high gene expression and H3K4 dimethylation and H4K5 acetylation, suggesting that they tend to occur in regions of open chromatin. The translocations likely result as a consequence of double-stranded break repair in the dihaploid genomes via homologous recombination during which IVP chromosomes are used as templates. Additionally, primary trisomy was observed in eight individuals. As the trisomic chromosomes were derived from Superior, meiotic nondisjunction may be common in potato., (© 2019 The Author(s).)
- Published
- 2019
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4. Reply to Huang et al.: Avoiding "one-size-fits-all" approaches to variant discovery.
- Author
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Hardigan MA, Laimbeer FPE, Hamilton JP, Vaillancourt B, Douches DS, Farré EM, Veilleux RE, and Buell CR
- Abstract
Competing Interests: The authors declare no conflict of interest.
- Published
- 2018
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5. Genome Reduction in Tetraploid Potato Reveals Genetic Load, Haplotype Variation, and Loci Associated With Agronomic Traits.
- Author
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Manrique-Carpintero NC, Coombs JJ, Pham GM, Laimbeer FPE, Braz GT, Jiang J, Veilleux RE, Buell CR, and Douches DS
- Abstract
The cultivated potato ( Solanum tuberosum ) has a complex genetic structure due to its autotetraploidy and vegetative propagation which leads to accumulation of mutations and a highly heterozygous genome. A high degree of heterozygosity has been considered to be the main driver of fitness and agronomic trait performance in potato improvement efforts, which is negatively impacted by genetic load. To understand the genetic landscape of cultivated potato, we constructed a gynogenic dihaploid (2 n = 2 x = 24) population from cv. Superior, prior to development of a high-density genetic map containing 12,753 single nucleotide polymorphisms (SNPs). Common quantitative trait loci (QTL) were identified for tuber traits, vigor and height on chromosomes 2, 4, 7, and 10, while specific QTL for number of inflorescences per plant, and tuber shape were present on chromosomes 4, 6, 10, and 11. Simplex rather than duplex loci were mainly associated with traits. In general, the Q allele (main effect) detected in one or two homologous chromosomes was associated with lower mean trait values suggesting the importance of dosage allelic effects, and the presence of up to two undesired alleles in the QTL region. Loss of heterozygosity has been associated with a lower rate of fitness, yet no correlation between the percent heterozygosity and increased fitness or agronomic performance was observed. Based upon linkage phase, we reconstructed the four homologous chromosome haplotypes of cv. Superior. revealing heterogeneity throughout the genome yet nearly duplicate haplotypes occurring among the homologs of particular chromosomes. These results suggest that the potentially deleterious mutations associated with genetic load in tetraploid potato could be mitigated by multiple loci which is consistent with the theory that epistasis complicates the identification of associations between markers and phenotypic performance.
- Published
- 2018
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6. Measuring Endoreduplication by Flow Cytometry of Isolated Tuber Protoplasts.
- Author
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Laimbeer FPE, Makris M, and Veilleux RE
- Subjects
- Endoreduplication genetics, Flow Cytometry methods, Protoplasts metabolism
- Abstract
Endoreduplication, the replication of a cell's nuclear genome without subsequent cytokinesis, yields cells with increased DNA content and is associated with specialization, development and increase in cellular size. In plants, endoreduplication seems to facilitate the growth and expansion of certain tissues and organs. Among them is the tuber of potato (Solanum tuberosum), which undergoes considerable cellular expansion in fulfilling its function of carbohydrate storage. Thus, endoreduplication may play an important role in how tubers are able to accommodate this abundance of carbon. However, the cellular debris resulting from crude nuclear isolation methods of tubers, methods that can be used effectively with leaves, precludes the estimation of the tuber endoreduplication index (EI). This article presents a technique for assessing tuber endoreduplication through the isolation of protoplasts while demonstrating representative results obtained from different genotypes and compartmentalized tuber tissues. The major limitations of the protocol are the time and reagent costs required for sample preparation as well as relatively short lifespan of samples after lysis of protoplasts. While the protocol is sensitive to technical variation, it represents an improvement over traditional methods of nuclear isolation from these large specialized cells. Possibilities for improvements to the protocol such as recycling enzyme, the use of fixatives, and other alterations are proposed.
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- 2018
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7. Lanosterol synthase-like is involved with differential accumulation of steroidal glycoalkaloids in potato.
- Author
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Kumar A, Fogelman E, Weissberg M, Tanami Z, Veilleux RE, and Ginzberg I
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- Amino Acid Sequence, Arabidopsis genetics, Biosynthetic Pathways, Genes, Reporter, Intramolecular Transferases genetics, Plants, Genetically Modified, Sequence Alignment, Solanum tuberosum genetics, Arabidopsis enzymology, Intramolecular Transferases metabolism, Phytosterols metabolism, Solanine metabolism, Solanum tuberosum enzymology, Triterpenes metabolism
- Abstract
Main Conclusion: Phytosterol homeostasis may be maintained in leaves through diversion of intermediates into glycoalkaloid biosynthesis, whereas in tuber flesh, excess intermediates are catalyzed by tuber-specific StLAS - like , resulting in low tuber glycoalkaloids. Lanosterol synthase (LAS) and cycloartenol synthase (CAS) are phylogenetically related enzymes. Cycloartenol is the accepted precursor leading to cholesterol and phytosterols, and in potato, to steroidal glycoalkaloid (SGA) biosynthesis. LAS was also shown to synthesize some plant sterols, albeit at trace amounts, questioning its role in sterol homeostasis. Presently, a potato LAS-related gene (StLAS-like) was identified and its activity verified in a yeast complementation assay. A transgenic approach with targeted gene expression and metabolic profiling of sterols and SGAs was used. Analyses of StLAS-like transcript levels and StLAS-like-promoter::GUS reporter assays indicated specific expression in tuber flesh tissue. Overexpression of Arabidopsis AtLAS in leaves where the endogenic StLAS-like is not expressed, resulted with increased SGA level and reduced phytosterol level, while in the tuber flesh SGA level was reduced. StLAS-like expression only in tuber flesh may explain the differential accumulation of SGAs in commercial cultivars-low in tubers, high in leaves. In leaves, to maintain phytosterol homeostasis, an excess of intermediates may be diverted into SGA biosynthesis, whereas in tuber flesh these intermediates are catalyzed by tuber-specific StLAS-like instead, resulting in low levels of SGA.
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- 2017
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8. Genome diversity of tuber-bearing Solanum uncovers complex evolutionary history and targets of domestication in the cultivated potato.
- Author
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Hardigan MA, Laimbeer FPE, Newton L, Crisovan E, Hamilton JP, Vaillancourt B, Wiegert-Rininger K, Wood JC, Douches DS, Farré EM, Veilleux RE, and Buell CR
- Subjects
- Alleles, Carbohydrate Metabolism genetics, Cell Cycle genetics, Chromosomes, Plant, Circadian Rhythm genetics, Diploidy, Endoreduplication genetics, Fertility genetics, Gametogenesis genetics, Gene Expression Regulation, Plant, Gene Pool, Genotype, Haplotypes, Metabolic Networks and Pathways genetics, North America, Peru, Phenotype, Phylogeny, Pollen genetics, Pollen growth & development, Polyploidy, South America, Species Specificity, Tetraploidy, Biological Evolution, Domestication, Genes, Plant genetics, Genetic Variation, Plant Tubers genetics, Solanum genetics, Solanum tuberosum genetics
- Abstract
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., Competing Interests: The authors declare no conflict of interest., (Copyright © 2017 the Author(s). Published by PNAS.)
- Published
- 2017
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9. Meiotic crossovers are associated with open chromatin and enriched with Stowaway transposons in potato.
- Author
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Marand AP, Jansky SH, Zhao H, Leisner CP, Zhu X, Zeng Z, Crisovan E, Newton L, Hamernik AJ, Veilleux RE, Buell CR, and Jiang J
- Subjects
- Chromosome Mapping, Chromosomes, Plant, Genomics, Haplotypes, Chromatin chemistry, Crossing Over, Genetic, DNA Transposable Elements, Meiosis genetics, Solanum tuberosum genetics
- Abstract
Background: Meiotic recombination is the foundation for genetic variation in natural and artificial populations of eukaryotes. Although genetic maps have been developed for numerous plant species since the late 1980s, few of these maps have provided the necessary resolution needed to investigate the genomic and epigenomic features underlying meiotic crossovers., Results: Using a whole genome sequencing-based approach, we developed two high-density reference-based haplotype maps using diploid potato clones as parents. The vast majority (81%) of meiotic crossovers were mapped to less than 5 kb. The fine-scale accuracy of crossover detection was validated by Sanger sequencing for a subset of ten crossover events. We demonstrate that crossovers reside in genomic regions of "open chromatin", which were identified based on hypersensitivity to DNase I digestion and association with H3K4me3-modified nucleosomes. The genomic regions spanning crossovers were significantly enriched with the Stowaway family of miniature inverted-repeat transposable elements (MITEs). The occupancy of Stowaway elements in gene promoters is concomitant with an increase in recombination rate. A generalized linear model identified the presence of Stowaway elements as the third most important genomic or chromatin feature behind genes and open chromatin for predicting crossover formation over 10-kb windows., Conclusions: Collectively, our results suggest that meiotic crossovers in potato are largely determined by the local chromatin status, marked by accessible chromatin, H3K4me3-modified nucleosomes, and the presence of Stowaway transposons.
- Published
- 2017
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10. Protoplast isolation prior to flow cytometry reveals clear patterns of endoreduplication in potato tubers, related species, and some starchy root crops.
- Author
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Laimbeer FPE, Holt SH, Makris M, Hardigan MA, Robin Buell C, and Veilleux RE
- Abstract
Background: Endoreduplication, the process of DNA replication in the absence of cell division, is associated with specialized cellular function and increased cell size. Genes controlling endoreduplication in tomato fruit have been shown to affect mature fruit size. An efficient method of estimating endoreduplication is required to study its role in plant organ development. Flow cytometry is often utilized to evaluate endoreduplication, yet some tissues and species, among them the tubers of Solanum tuberosum , remain intractable to routine tissue preparation for flow cytometry. We aimed to develop a method through the use of protoplast extraction preceding flow cytometry, specifically for the assessment of endoreduplication in potato tubers., Results: We present a method for appraising endoreduplication in potato ( Solanum tuberosum ) tuber tissues. We evaluated this method and observed consistent differences between pith and cortex of tubers and between different cultivars, but no apparent relationship with whole tuber size. Furthermore, we were able to observe distinct patterns of endoreduplication in 16 of 20 wild potato relatives, with mean endoreduplication index (EI) ranging from 0.94 to 2.62 endocycles per cell. The protocol was also applied to a panel of starchy root crop species and, while only two of five yielded reliable flow histograms, the two (sweet potato and turnip) exhibited substantially lower EIs than wild and cultivated potato accessions., Conclusions: The protocol reported herein has proven effective on tubers of a variety of potato cultivars and related species, as well as storage roots of other starchy crops. This method provides an important tool for the study of potato morphology and development while revealing natural variation for endoreduplication which may have agricultural relevance.
- Published
- 2017
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11. Self-Fertility in a Cultivated Diploid Potato Population Examined with the Infinium 8303 Potato Single-Nucleotide Polymorphism Array.
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Peterson BA, Holt SH, Laimbeer FP, Doulis AG, Coombs J, Douches DS, Hardigan MA, Buell CR, and Veilleux RE
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- Chromosome Mapping, DNA Copy Number Variations, Diploidy, Fertility genetics, Genotype, Oligonucleotide Array Sequence Analysis, Polymorphism, Single Nucleotide, Genome, Plant genetics, Solanum tuberosum genetics
- Abstract
Within a population of F hybrids between two genotypes ( L. Group Phureja DM 1-3 516 R44 [DM] and L. Group Tuberosum RH89-039-16 [RH]) used in the potato genome sequencing project, we observed fruit set after self-pollination on many plants. Examination of pollen tube growth in self-fertile and self-unfruitful F plants after controlled self-pollinations revealed no difference in the ability of pollen tubes to reach the ovary. To identify genomic regions linked with self-fertility, we genotyped the F population using a genome-wide single-nucleotide polymorphism (SNP) array. Polymorphic and robust SNPs were analyzed to identify allelic states segregating with the self-fertile phenotype. All 88 highly significant SNPs occurred on chromosome 12. Seeds obtained after self-pollination of self-fertile individuals were used to advance the population for four generations. Genotyping 46 self-fruitful and 46 self-unfruitful S plants on the Infinium 8303 Potato SNP array revealed eight SNPs segregating with self-fertility on chromosomes 4, 9, 11, and 12. Three times more heterozygosity than expected was found in the S generation. Estimates of heterozygosity were influenced by copy number variation (CNV) in the potato genome leading to spurious heterozygous genotyping calls. Some spurious heterozygosity could be removed by application of a CNV filter developed from alignment of additional monoploid potato genomic sequence to the DM reference genome. The genes responsible for fruit set in self-fertile plants in the F generation were restricted to chromosome 12, whereas new genomic regions contributed to the ability of S plants to set fruit after self-pollination., (Copyright © 2016 Crop Science Society of America.)
- Published
- 2016
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12. Comparative Analysis of Regions with Distorted Segregation in Three Diploid Populations of Potato.
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Manrique-Carpintero NC, Coombs JJ, Veilleux RE, Buell CR, and Douches DS
- Subjects
- Chromosome Segregation, Chromosomes, Plant, Epistasis, Genetic, Genotype, Meiosis, Recombination, Genetic, Self-Incompatibility in Flowering Plants genetics, Chromosome Mapping, Diploidy, Polymorphism, Single Nucleotide, Solanum tuberosum genetics
- Abstract
Genes associated with gametic and zygotic selection could underlie segregation distortion, observed as alterations of expected Mendelian genotypic frequencies in mapping populations. We studied highly dense genetic maps based on single nucleotide polymorphisms to elucidate the genetic nature of distorted segregation in potato. Three intra- and interspecific diploid segregating populations were used. DRH and D84 are crosses between the sequenced doubled monoploid DM 1-3 516 R44 Solanum tuberosum Group Phureja and either RH89-039-16 S. tuberosum or 84SD22, a S. tuberosum × S. chacoense hybrid. MSX902 is an interspecific cross between 84SD22 and Ber83 S. berthaultii × 2 × species mosaic. At the 0.05 significance level, 21%, 57%, and 51% of the total markers mapped in DRH, D84, and MSX902 exhibited distorted segregation, respectively. Segregation distortion regions for DRH were located on chromosomes 9 and 12; for D84 on chromosomes 2, 3, 4, 6, 7, and 8; and on chromosomes 1, 2, 7, 9, and 12 for MSX902. In general, each population had unique segregation distortion regions and directions of distortion. Interspecific crosses showed greater levels of distorted segregation and lower recombination rates as determined from the male parents. The different genomic regions where the segregation distortion regions occurred in the three populations likely reflect unique genetic combinations producing distorted segregation., (Copyright © 2016 Manrique-Carpintero et al.)
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- 2016
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13. Genome Reduction Uncovers a Large Dispensable Genome and Adaptive Role for Copy Number Variation in Asexually Propagated Solanum tuberosum.
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Hardigan MA, Crisovan E, Hamilton JP, Kim J, Laimbeer P, Leisner CP, Manrique-Carpintero NC, Newton L, Pham GM, Vaillancourt B, Yang X, Zeng Z, Douches DS, Jiang J, Veilleux RE, and Buell CR
- Subjects
- Genotype, In Situ Hybridization, Fluorescence, Phenotype, Phylogeny, Reproduction, Asexual genetics, Solanum tuberosum physiology, Species Specificity, DNA Copy Number Variations genetics, Gene Duplication, Genetic Variation, Genome, Plant genetics, Plant Proteins genetics, Solanum tuberosum genetics
- Abstract
Clonally reproducing plants have the potential to bear a significantly greater mutational load than sexually reproducing species. To investigate this possibility, we examined the breadth of genome-wide structural variation in a panel of monoploid/doubled monoploid clones generated from native populations of diploid potato (Solanum tuberosum), a highly heterozygous asexually propagated plant. As rare instances of purely homozygous clones, they provided an ideal set for determining the degree of structural variation tolerated by this species and deriving its minimal gene complement. Extensive copy number variation (CNV) was uncovered, impacting 219.8 Mb (30.2%) of the potato genome with nearly 30% of genes subject to at least partial duplication or deletion, revealing the highly heterogeneous nature of the potato genome. Dispensable genes (>7000) were associated with limited transcription and/or a recent evolutionary history, with lower deletion frequency observed in genes conserved across angiosperms. Association of CNV with plant adaptation was highlighted by enrichment in gene clusters encoding functions for environmental stress response, with gene duplication playing a part in species-specific expansions of stress-related gene families. This study revealed unique impacts of CNV in a species with asexual reproductive habits and how CNV may drive adaption through evolution of key stress pathways., (© 2016 American Society of Plant Biologists. All rights reserved.)
- Published
- 2016
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14. Characterization of a potato activation-tagged mutant, nikku, and its partial revertant.
- Author
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Aulakh SS, Veilleux RE, Tang G, and Flinn BS
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- 5-Methylcytosine metabolism, Biological Assay, Blotting, Southern, Cytidine analogs & derivatives, Cytidine pharmacology, DNA Methylation drug effects, DNA Methylation genetics, DNA, Bacterial genetics, Enhancer Elements, Genetic genetics, Gene Dosage, Gene Expression Regulation, Plant drug effects, Phenotype, Plant Tubers drug effects, Plant Tubers physiology, RNA, Messenger genetics, RNA, Messenger metabolism, Solanum tuberosum drug effects, Solanum tuberosum growth & development, Stress, Physiological drug effects, Mutagenesis genetics, Mutation genetics, Solanum tuberosum genetics
- Abstract
Main Conclusion: A potato mutant with a strong stress-response phenotype, and a partial mutant revertant, were characterized. Gene expression patterns and DNA cytosine methylation varied between these and wild-type, indicating a role for DNA cytosine methylation changes in the gene expression and visible phenotypes. Morphological and molecular studies were conducted to compare potato cv. Bintje, a Bintje activation-tagged mutant (nikku), and nikku revertant phenotype plants. Morphological studies revealed that nikku plants exhibited an extremely dwarf phenotype, had small hyponastic leaves, were rootless, and infrequently produced small tubers compared to wild-type Bintje. The overall phenotype was suggestive of a constitutive stress response, which was further supported by the greater expression level of several stress-responsive genes in nikku. Unlike the nikku mutant, the revertant exhibited near normal shoot elongation, larger leaves and consistent rooting. The reversion appeared partial, and was not the result of a loss of 35S enhancer copies from the original nikku mutant. Southern blot analyses indicated the presence of a single T-DNA insertion on chromosome 12 in the mutant. Gene expression studies comparing Bintje, nikku and revertant phenotype plants indicated transcriptional activation/repression of several genes flanking both sides of the insertion in the mutant, suggesting that activation tagging had pleiotropic effects in nikku. In contrast, gene expression levels for many, but not all, of the same genes in the revertant were similar to Bintje, indicating some reversion at the gene expression level as well. DNA methylation studies indicated differences in cytosine methylation status of the 35S enhancers between the nikku mutant and its revertant. In addition, global DNA cytosine methylation varied between Bintje, the nikku mutant and the revertant, suggesting involvement in gene expression changes, as well as mutant phenotype.
- Published
- 2015
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15. Transposon based activation tagging in diploid strawberry and monoploid derivatives of potato.
- Author
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Lu N, Carter JD, Boluarte Medina T, Holt SH, Manrique-Carpintero NC, Upham KT, Pereira A, Shulaev V, and Veilleux RE
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- Agrobacterium genetics, Base Sequence, Crops, Agricultural genetics, Germination, Molecular Sequence Data, Mutation, Plants, Genetically Modified, Polymerase Chain Reaction methods, Transformation, Genetic, DNA Transposable Elements, Diploidy, Fragaria genetics, Solanum tuberosum genetics
- Abstract
Key Message: Diploid strawberry and potato transformed with a transposon tagging construct exhibited either global (strawberry) or local transposition (potato). An activation tagged, compact-sized strawberry mutant overexpressed the gene adjacent to Ds. As major fruit and vegetable crops, respectively, strawberry and potato are among the first horticultural crops with draft genome sequences. To study gene function, we examined transposon-tagged mutant strategies in model populations for both species, Fragaria vesca and Solanum tuberosum Group Phureja, using the same Activation/Dissociation (Ac/Ds) construct. Early somatic transposition during tissue culture occurred at a frequency of 18.5% in strawberry but not in potato transformants. Green fluorescent protein under a monocot promoter was a more reliable selectable marker in strawberry compared to potato. BASTA (gluphosinate herbicide) resistance served as an effective selectable marker for both species (80 and 85% reliable in strawberry and potato, respectively), although the effective concentration differed (0.5% for strawberry and 0.03% for potato). Transposons preferentially reinserted within genes (exons and introns) in both species. Real-time quantitative PCR revealed enhanced gene expression (670 and 298-fold expression compared to wild type in petiole and leaf tissue, respectively) for an activation tagged strawberry mutant with Ds inserted about 0.6 kb upstream from a gene coding for an epidermis-specific secreted glycoprotein EP1. Our data also suggested that endopolyploid (diploid) cells occurring in leaf explants of monoploid potato were the favored targets of T-DNA integration during transformation. Mutants obtained in these studies provide a useful resource for future genetic studies.
- Published
- 2014
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16. Characterization and RNA-seq analysis of underperformer, an activation-tagged potato mutant.
- Author
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Aulakh SS, Veilleux RE, Dickerman AW, Tang G, and Flinn BS
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- Amino Acid Sequence, Base Sequence, Computational Biology, Flowers genetics, Flowers growth & development, Gene Expression Profiling, High-Throughput Nucleotide Sequencing, Molecular Sequence Data, Mutagenesis, Insertional, Phenotype, Plant Leaves genetics, Plant Leaves growth & development, Plant Stems genetics, Plant Stems growth & development, Plant Tubers genetics, Plant Tubers growth & development, RNA, Messenger chemistry, RNA, Messenger genetics, RNA, Plant chemistry, RNA, Plant genetics, Sequence Analysis, RNA, Solanum tuberosum growth & development, Gene Expression Regulation, Plant, Solanum tuberosum genetics, Transcriptome
- Abstract
The potato cv. Bintje and a Bintje activation-tagged mutant, underperformer (up) were compared. Mutant up plants grown in vitro were dwarf, with abundant axillary shoot growth, greater tuber yield, altered tuber traits and early senescence compared to wild type. Under in vivo conditions, the dwarf and early senescence phenotypes of the mutant remained, but the up plants exhibited a lower tuber yield and fewer axillary shoots compared to wild type. Southern blot analyses indicated a single T-DNA insertion in the mutant, located on chromosome 10. Initial PCR-based gene expression studies indicated transcriptional activation/repression of several genes in the mutant flanking the insertion. The gene immediately flanking the right border of the T-DNA insertion, which encoded an uncharacterized Broad complex, Tramtrac, Bric-a-brac; also known as Pox virus and Zinc finger (BTB/POZ) domain-containing protein (StBTB/POZ1) containing an Armadillo repeat region, was up-regulated in the mutant. Global gene expression comparisons between Bintje and up using RNA-seq on leaves from 60 day-old plants revealed a dataset of over 1,600 differentially expressed genes. Gene expression analyses suggested a variety of biological processes and pathways were modified in the mutant, including carbohydrate and lipid metabolism, cell division and cell cycle activity, biotic and abiotic stress responses, and proteolysis.
- Published
- 2014
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17. Allelic variation in genes contributing to glycoalkaloid biosynthesis in a diploid interspecific population of potato.
- Author
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Manrique-Carpintero NC, Tokuhisa JG, Ginzberg I, and Veilleux RE
- Subjects
- Genes, Plant, Polymorphism, Single Nucleotide, Alkaloids biosynthesis, Alleles, Diploidy, Solanum tuberosum genetics
- Abstract
Key Message: Variation for allelic state within genes of both primary and secondary metabolism influences the quantity and quality of steroidal glycoalkaloids produced in potato leaves. Genetic factors associated with the biosynthesis and accumulation of steroidal glycoalkaloids (SGAs) in potato were addressed by a candidate gene approach and whole genome single nucleotide polymorphism (SNP) genotyping. Allelic sequences spanning coding regions of four candidate genes [3-hydroxy-3-methylglutaryl coenzyme A reductase 2 (HMG2); 2,3-squalene epoxidase; solanidine galactosyltransferase; and solanidine glucosyltransferase (SGT2)] were obtained from two potato species differing in SGA composition: Solanum chacoense (chc 80-1) and Solanum tuberosum group Phureja (phu DH). An F2 population was genotyped and foliar SGAs quantified. The concentrations of α-solanine, α-chaconine, leptine I, leptine II and total SGAs varied broadly among F2 individuals. F2 plants with chc 80-1 alleles for HMG2 or SGT2 accumulated significantly greater leptines and total SGAs compared to plants with phu DH alleles. Plants with chc 80-1 alleles at both loci expressed the greatest levels of total SGAs, α-solanine and α-chaconine. A significant positive correlation was found between α-solanine and α-chaconine accumulation as well as between leptine I and leptine II. A whole genome SNP genotyping analysis of an F2 subsample verified the importance of chc 80-1 alleles at HMG2 and SGT2 for SGA synthesis and accumulation and suggested additional candidate genes including some previously associated with SGA production. Loci on five and seven potato pseudochromosomes were associated with synthesis and accumulation of SGAs, respectively. Two loci, on pseudochromosomes 1 and 6, explained phenotypic segregation of α-solanine and α-chaconine synthesis. Knowledge of the genetic factors influencing SGA production in potato may assist breeding for pest resistance.
- Published
- 2014
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18. Construction of reference chromosome-scale pseudomolecules for potato: integrating the potato genome with genetic and physical maps.
- Author
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Sharma SK, Bolser D, de Boer J, Sønderkær M, Amoros W, Carboni MF, D'Ambrosio JM, de la Cruz G, Di Genova A, Douches DS, Eguiluz M, Guo X, Guzman F, Hackett CA, Hamilton JP, Li G, Li Y, Lozano R, Maass A, Marshall D, Martinez D, McLean K, Mejía N, Milne L, Munive S, Nagy I, Ponce O, Ramirez M, Simon R, Thomson SJ, Torres Y, Waugh R, Zhang Z, Huang S, Visser RG, Bachem CW, Sagredo B, Feingold SE, Orjeda G, Veilleux RE, Bonierbale M, Jacobs JM, Milbourne D, Martin DM, and Bryan GJ
- Subjects
- Biomarkers metabolism, Chromosomes, Plant metabolism, Genome, Plant, Internet, User-Computer Interface, Chromosome Mapping standards, Chromosomes, Plant genetics, Solanum tuberosum genetics
- Abstract
The genome of potato, a major global food crop, was recently sequenced. The work presented here details the integration of the potato reference genome (DM) with a new sequence-tagged site marker-based linkage map and other physical and genetic maps of potato and the closely related species tomato. Primary anchoring of the DM genome assembly was accomplished by the use of a diploid segregating population, which was genotyped with several types of molecular genetic markers to construct a new ~936 cM linkage map comprising 2469 marker loci. In silico anchoring approaches used genetic and physical maps from the diploid potato genotype RH89-039-16 (RH) and tomato. This combined approach has allowed 951 superscaffolds to be ordered into pseudomolecules corresponding to the 12 potato chromosomes. These pseudomolecules represent 674 Mb (~93%) of the 723 Mb genome assembly and 37,482 (~96%) of the 39,031 predicted genes. The superscaffold order and orientation within the pseudomolecules are closely collinear with independently constructed high density linkage maps. Comparisons between marker distribution and physical location reveal regions of greater and lesser recombination, as well as regions exhibiting significant segregation distortion. The work presented here has led to a greatly improved ordering of the potato reference genome superscaffolds into chromosomal "pseudomolecules".
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- 2013
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19. Sequence diversity in coding regions of candidate genes in the glycoalkaloid biosynthetic pathway of wild potato species.
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Manrique-Carpintero NC, Tokuhisa JG, Ginzberg I, Holliday JA, and Veilleux RE
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- Alkaloids genetics, Alleles, Galactosyltransferases genetics, Genotype, Glucosyltransferases chemistry, Glucosyltransferases genetics, HMGB1 Protein genetics, HMGB2 Protein genetics, Open Reading Frames, Polymorphism, Single Nucleotide, Protein Structure, Tertiary, Sequence Analysis, DNA, Squalene Monooxygenase genetics, Alkaloids biosynthesis, Genome, Plant, Solanum tuberosum genetics
- Abstract
Natural variation in five candidate genes of the steroidal glycoalkaloid (SGA) metabolic pathway and whole-genome single nucleotide polymorphism (SNP) genotyping were studied in six wild [Solanum chacoense (chc 80-1), S. commersonii, S. demissum, S. sparsipilum, S. spegazzinii, S. stoloniferum] and cultivated S. tuberosum Group Phureja (phu DH) potato species with contrasting levels of SGAs. Amplicons were sequenced for five candidate genes: 3-hydroxy-3-methylglutaryl coenzyme A reductase 1 and 2 (HMG1, HMG2) and 2.3-squalene epoxidase (SQE) of primary metabolism, and solanidine galactosyltransferase (SGT1), and glucosyltransferase (SGT2) of secondary metabolism. SNPs (n = 337) producing 354 variations were detected within 3.7 kb of sequenced DNA. More polymorphisms were found in introns than exons and in genes of secondary compared to primary metabolism. Although no significant deviation from neutrality was found, dN/dS ratios < 1 and negative values of Tajima's D test suggested purifying selection and genetic hitchhiking in the gene fragments. In addition, patterns of dN/dS ratios across the SGA pathway suggested constraint by natural selection. Comparison of nucleotide diversity estimates and dN/dS ratios showed stronger selective constraints for genes of primary rather than secondary metabolism. SNPs (n = 24) with an exclusive genotype for either phu DH (low SGA) or chc 80-1 (high SGA) were identified for HMG2, SQE, SGT1 and SGT2. The SolCAP 8303 Illumina Potato SNP chip genotyping revealed eight informative SNPs on six pseudochromosomes, with homozygous and heterozygous genotypes that discriminated high, intermediate and low levels of SGA accumulation. These results can be used to evaluate SGA accumulation in segregating or association mapping populations.
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- 2013
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20. Retrospective view of North American potato (Solanum tuberosum L.) breeding in the 20th and 21st centuries.
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Hirsch CN, Hirsch CD, Felcher K, Coombs J, Zarka D, Van Deynze A, De Jong W, Veilleux RE, Jansky S, Bethke P, Douches DS, and Buell CR
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- Agriculture, Biosynthetic Pathways genetics, Carotenoids metabolism, Gene Frequency genetics, Geography, Heterozygote, History, 20th Century, History, 21st Century, Marketing, North America, Phenotype, Polymorphism, Single Nucleotide genetics, Seeds genetics, Selection, Genetic, Solanum tuberosum economics, Solanum tuberosum genetics, Breeding history, Solanum tuberosum growth & development, Solanum tuberosum history
- Abstract
Cultivated potato (Solanum tuberosum L.), a vegetatively propagated autotetraploid, has been bred for distinct market classes, including fresh market, pigmented, and processing varieties. Breeding efforts have relied on phenotypic selection of populations developed from intra- and intermarket class crosses and introgressions of wild and cultivated Solanum relatives. To retrospectively explore the effects of potato breeding at the genome level, we used 8303 single-nucleotide polymorphism markers to genotype a 250-line diversity panel composed of wild species, genetic stocks, and cultivated potato lines with release dates ranging from 1857 to 2011. Population structure analysis revealed four subpopulations within the panel, with cultivated potato lines grouping together and separate from wild species and genetic stocks. With pairwise kinship estimates clear separation between potato market classes was observed. Modern breeding efforts have scarcely changed the percentage of heterozygous loci or the frequency of homozygous, single-dose, and duplex loci on a genome level, despite concerted efforts by breeders. In contrast, clear selection in less than 50 years of breeding was observed for alleles in biosynthetic pathways important for market class-specific traits such as pigmentation and carbohydrate composition. Although improvement and diversification for distinct market classes was observed through whole-genome analysis of historic and current potato lines, an increased rate of gain from selection will be required to meet growing global food demands and challenges due to climate change. Understanding the genetic basis of diversification and trait improvement will allow for more rapid genome-guided improvement of potato in future breeding efforts.
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- 2013
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21. An active ac/ds transposon system for activation tagging in tomato cultivar m82 using clonal propagation.
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Carter JD, Pereira A, Dickerman AW, and Veilleux RE
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- DNA, Plant genetics, Databases, Nucleic Acid, Genome, Plant genetics, Solanum lycopersicum physiology, Mutagenesis, Insertional, Phenotype, Plant Proteins genetics, Plant Proteins metabolism, Plants, Genetically Modified, Seeds genetics, Seeds physiology, Sequence Analysis, DNA, Transposases metabolism, Zea mays genetics, DNA Transposable Elements genetics, Solanum lycopersicum genetics, Transposases genetics
- Abstract
Tomato (Solanum lycopersicum) is a model organism for Solanaceae in both molecular and agronomic research. This project utilized Agrobacterium tumefaciens transformation and the transposon-tagging construct Activator (Ac)/Dissociator (Ds)-ATag-Bar_gosGFP to produce activation-tagged and knockout mutants in the processing tomato cultivar M82. The construct carried hygromycin resistance (hyg), green fluorescent protein (GFP), and the transposase (TPase) of maize (Zea mays) Activator major transcript X054214.1 on the stable Ac element, along with a 35S enhancer tetramer and glufosinate herbicide resistance (BAR) on the mobile Ds-ATag element. An in vitro propagation strategy was used to produce a population of 25 T0 plants from a single transformed plant regenerated in tissue culture. A T1 population of 11,000 selfed and cv M82 backcrossed progeny was produced from the functional T0 line. This population was screened using glufosinate herbicide, hygromycin leaf painting, and multiplex polymerase chain reaction (PCR). Insertion sites of transposed Ds-ATag elements were identified through thermal asymmetric interlaced PCR, and resulting product sequences were aligned to the recently published tomato genome. A population of 509 independent, Ds-only transposant lines spanning all 12 tomato chromosomes has been developed. Insertion site analysis demonstrated that more than 80% of these lines harbored Ds insertions conducive to activation tagging. The capacity of the Ds-ATag element to alter transcription was verified by quantitative real-time reverse transcription-PCR in two mutant lines. The transposon-tagged lines have been immortalized in seed stocks and can be accessed through an online database, providing a unique resource for tomato breeding and analysis of gene function in the background of a commercial tomato cultivar.
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- 2013
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22. Development of an efficient transformation method by Agrobacterium tumefaciens and high throughput spray assay to identify transgenic plants for woodland strawberry (Fragaria vesca) using NPTII selection.
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Pantazis CJ, Fisk S, Mills K, Flinn BS, Shulaev V, Veilleux RE, and Dan Y
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- Acetophenones pharmacology, DNA, Bacterial genetics, Fragaria drug effects, Fragaria growth & development, Genome Size, Genomics, Germination, High-Throughput Screening Assays, Kanamycin pharmacology, Mutagenesis, Insertional, Plant Leaves drug effects, Plant Leaves genetics, Plant Leaves growth & development, Plant Roots drug effects, Plant Roots genetics, Plant Roots growth & development, Plant Shoots drug effects, Plant Shoots genetics, Plant Shoots growth & development, Plants, Genetically Modified, Regeneration, Seeds drug effects, Seeds genetics, Seeds growth & development, Transgenes, Agrobacterium tumefaciens genetics, Anti-Bacterial Agents pharmacology, Fragaria genetics, Paromomycin pharmacology, Transformation, Genetic drug effects
- Abstract
KEY MESSAGE : We developed an efficient Agrobacterium -mediated transformation method using an Ac/Ds transposon tagging construct for F. vesca and high throughput paromomycin spray assay to identify its transformants for strawberry functional genomics. Genomic resources for Rosaceae species are now readily available, including the Fragaria vesca genome, EST sequences, markers, linkage maps, and physical maps. The Rosaceae Genomic Executive Committee has promoted strawberry as a translational genomics model due to its unique biological features and transformability for fruit trait improvement. Our overall research goal is to use functional genomic and metabolic approaches to pursue high throughput gene discovery in the diploid woodland strawberry. F. vesca offers several advantages of a fleshy fruit typical of most fruit crops, short life cycle (seed to seed in 12-16 weeks), small genome size (206 Mbb/C), small plant size, self-compatibility, and many seeds per plant. We have developed an efficient Agrobacterium tumefaciens-mediated strawberry transformation method using kanamycin selection, and high throughput paromomycin spray assay to efficiently identify transgenic strawberry plants. Using our kanamycin transformation method, we were able to produce up to 98 independent kanamycin resistant insertional mutant lines using a T-DNA construct carrying an Ac/Ds transposon Launchpad system from a single transformation experiment involving inoculation of 22 leaf explants of F. vesca accession 551572 within approx. 11 weeks (from inoculation to soil). Transgenic plants with 1-2 copies of a transgene were confirmed by Southern blot analysis. Using our paromomycin spray assay, transgenic F. vesca plants were rapidly identified within 10 days after spraying.
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- 2013
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23. Transposon tagging in diploid strawberry.
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Veilleux RE, Mills KP, Baxter AJ, Upham KT, Ferguson TJ, Holt SH, Lu N, Ruiz-Rojas JJ, Pantazis CJ, Davis CM, Lindsay RC, Powell FL, Dan Y, Dickerman AW, Oosumi T, and Shulaev V
- Subjects
- Diploidy, Genetic Engineering, Sequence Tagged Sites, Transformation, Genetic, Crops, Agricultural genetics, DNA Transposable Elements, Fragaria genetics, Plants, Genetically Modified genetics, Zea mays genetics
- Abstract
Fragaria vesca was transformed with a transposon tagging construct harbouring amino terminally deleted maize transposase and EGFP (Ac element), NPTII, CaMV 35S promoter (P35S) driving transposase and mannopine synthase promoter (Pmas) driving EGFP (Ds element). Of 180 primary transgenics, 48 were potential launch pads, 72 were multiple insertions or chimaeras, and 60 exhibited somatic transposition. T₁ progeny of 32 putative launch pads were screened by multiplex PCR for transposition. Evidence of germ-line transposition occurred in 13 putative launch pads; however, the transposition frequency was too low in three for efficient recovery of transposants. The transposition frequency in the remaining launch pads ranged from 16% to 40%. After self-pollination of the T₀ launch pads, putative transposants in the T₁ generation were identified by multiplex PCR. Sequencing of hiTAIL-PCR products derived from nested primers within the Ds end sequences (either P35S at the left border or the inverted repeat at the right border) of T₁ plants revealed transposition of the Ds element to distant sites in the strawberry genome. From more than 2400 T₁ plants screened, 103 unique transposants have been identified, among which 17 were somatic transpositions observed in the T₀ generation. Ds insertion sites were dispersed among various gene elements [exons (15%), introns (23%), promoters (30%), 3' UTRs (17%) as well as intergenically (15%)]. Three-primer (one on either side of the Ds insertion and one within the Ds T-DNA) PCR could be used to identify homozygous T₂ transposon-tagged plants. The mutant collection has been catalogued in an on-line database., (© 2012 The Authors. Plant Biotechnology Journal © 2012 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.)
- Published
- 2012
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24. Induction of potato steroidal glycoalkaloid biosynthetic pathway by overexpression of cDNA encoding primary metabolism HMG-CoA reductase and squalene synthase.
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Ginzberg I, Thippeswamy M, Fogelman E, Demirel U, Mweetwa AM, Tokuhisa J, and Veilleux RE
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- Chromatography, High Pressure Liquid, Farnesyl-Diphosphate Farnesyltransferase metabolism, Gene Expression Regulation, Plant, Genes, Plant genetics, Hydroxymethylglutaryl CoA Reductases metabolism, Phytosterols biosynthesis, Plant Leaves enzymology, Plant Leaves genetics, Plants, Genetically Modified, RNA, Messenger genetics, RNA, Messenger metabolism, Sequence Homology, Nucleic Acid, Solanine metabolism, Alkaloids biosynthesis, Biosynthetic Pathways genetics, DNA, Complementary genetics, Farnesyl-Diphosphate Farnesyltransferase genetics, Hydroxymethylglutaryl CoA Reductases genetics, Solanine analogs & derivatives, Solanum tuberosum enzymology, Solanum tuberosum genetics
- Abstract
Potato steroidal glycoalkaloids (SGAs) are toxic secondary metabolites whose total content in tubers must be regulated. SGAs are biosynthesized by the sterol branch of the mevalonic acid/isoprenoid pathway. In a previous study, we showed a correlation between SGA levels and the abundance of transcript coding for HMG-CoA reductase 1 (HMG1) and squalene synthase 1 (SQS1) in potato tissues and potato genotypes varying in SGA content. Here, Solanum tuberosum cv. Desirée (low SGA producer) was transformed with a gene construct containing the coding region of either HMG1 or SQS1 of Solanum chacoense Bitt. clone 8380-1, a high SGA producer. SGA levels in transgenic HMG-plants were either greater than (in eight of 14 plants) or no different from untransformed controls, whereas only four of 12 SQS-transgenics had greater SGA levels than control, as determined by HPLC. Quantitative real-time PCR was used to estimate relative steady-state transcript levels of isoprenoid-, steroid-, and SGA-related genes in leaves of the transgenic plants compared to nontransgenic controls. HMG-transgenic plants exhibited increased transcript accumulation of SQS1, sterol C24-methyltransferase type1 (SMT1), and solanidine glycosyltransferase 2 (SGT2), whereas SQS-transgenic plants, had consistently lower transcript levels of HMG1 and variable SMT1 and SGT2 transcript abundance among different transgenics. HMG-transgenic plants exhibited changes in transcript accumulation for some sterol biosynthetic genes as well. Taken together, the data suggest coordinated regulation of isoprenoid metabolism and SGA secondary metabolism.
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- 2012
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25. Steroidal glycoalkaloids in Solanum chacoense.
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Mweetwa AM, Hunter D, Poe R, Harich KC, Ginzberg I, Veilleux RE, and Tokuhisa JG
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- Molecular Conformation, Plant Extracts analysis, Plant Extracts metabolism, Plant Roots chemistry, Plant Roots metabolism, Solanaceous Alkaloids analysis, Solanum tuberosum chemistry, Solanum tuberosum growth & development, Stereoisomerism, Solanaceous Alkaloids metabolism, Solanum tuberosum metabolism
- Abstract
Potato (Solanum tuberosum L.), a domesticated species that is the fourth most important world agricultural commodity, requires significant management to minimize the effects of herbivore and pathogen damage on crop yield. A wild relative, Solanum chacoense Bitt., has been of interest to plant breeders because it produces an abundance of novel steroidal glycoalkaloid compounds, leptines and leptinines, which are particularly effective deterrents of herbivory by the Colorado potato beetle (Leptinotarsa decemlineata Say). Biochemical approaches were used in this study to investigate the formation and accumulation of SGAs in S. chacoense. SGA contents were determined in various organs at different stages of organ maturity during a time course of plant development. Leptines and leptinines were the main contributors to the increased levels in SGA concentration measured in the aerial versus the subterranean organs of S. chacoense accession 8380-1. Leptines were not detected in aboveground stolons until the stage where shoots had formed mature chlorophyllous leaves. To gain insights into SGA biosynthesis, the abundance of SGAs and steady-state transcripts of genes coding for enzymes of the central terpene and SGA-specific pathways in various plant organs at anthesis were compared. For two genes of primary terpene metabolism, transcript and SGA abundances were correlated, although with some discrepancies. For genes associated with SGA biosynthesis, transcripts were not detected in some tissues containing SGAs; however these transcripts were detected in the progenitor tissues, indicating the possibility that under our standard growth conditions, SGA biosynthesis is largely limited to highly proliferative tissues such as shoot, root and floral meristems., (Copyright © 2011 Elsevier Ltd. All rights reserved.)
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- 2012
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26. Integration of two diploid potato linkage maps with the potato genome sequence.
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Felcher KJ, Coombs JJ, Massa AN, Hansey CN, Hamilton JP, Veilleux RE, Buell CR, and Douches DS
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- Genotyping Techniques, Polymorphism, Single Nucleotide genetics, Reproducibility of Results, Chromosome Mapping methods, Diploidy, Genome, Plant genetics, Solanum tuberosum genetics
- Abstract
To facilitate genome-guided breeding in potato, we developed an 8303 Single Nucleotide Polymorphism (SNP) marker array using potato genome and transcriptome resources. To validate the Infinium 8303 Potato Array, we developed linkage maps from two diploid populations (DRH and D84) and compared these maps with the assembled potato genome sequence. Both populations used the doubled monoploid reference genotype DM1-3 516 R44 as the female parent but had different heterozygous diploid male parents (RH89-039-16 and 84SD22). Over 4,400 markers were mapped (1,960 in DRH and 2,454 in D84, 787 in common) resulting in map sizes of 965 (DRH) and 792 (D84) cM, covering 87% (DRH) and 88% (D84) of genome sequence length. Of the mapped markers, 33.5% were in candidate genes selected for the array, 4.5% were markers from existing genetic maps, and 61% were selected based on distribution across the genome. Markers with distorted segregation ratios occurred in blocks in both linkage maps, accounting for 4% (DRH) and 9% (D84) of mapped markers. Markers with distorted segregation ratios were unique to each population with blocks on chromosomes 9 and 12 in DRH and 3, 4, 6 and 8 in D84. Chromosome assignment of markers based on linkage mapping differed from sequence alignment with the Potato Genome Sequencing Consortium (PGSC) pseudomolecules for 1% of the mapped markers with some disconcordant markers attributable to paralogs. In total, 126 (DRH) and 226 (D84) mapped markers were not anchored to the pseudomolecules and provide new scaffold anchoring data to improve the potato genome assembly. The high degree of concordance between the linkage maps and the pseudomolecules demonstrates both the quality of the potato genome sequence and the functionality of the Infinium 8303 Potato Array. The broad genome coverage of the Infinium 8303 Potato Array compared to other marker sets will enable numerous downstream applications.
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- 2012
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27. Genome sequence and analysis of the tuber crop potato.
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Xu X, Pan S, Cheng S, Zhang B, Mu D, Ni P, Zhang G, Yang S, Li R, Wang J, Orjeda G, Guzman F, Torres M, Lozano R, Ponce O, Martinez D, De la Cruz G, Chakrabarti SK, Patil VU, Skryabin KG, Kuznetsov BB, Ravin NV, Kolganova TV, Beletsky AV, Mardanov AV, Di Genova A, Bolser DM, Martin DM, Li G, Yang Y, Kuang H, Hu Q, Xiong X, Bishop GJ, Sagredo B, Mejía N, Zagorski W, Gromadka R, Gawor J, Szczesny P, Huang S, Zhang Z, Liang C, He J, Li Y, He Y, Xu J, Zhang Y, Xie B, Du Y, Qu D, Bonierbale M, Ghislain M, Herrera Mdel R, Giuliano G, Pietrella M, Perrotta G, Facella P, O'Brien K, Feingold SE, Barreiro LE, Massa GA, Diambra L, Whitty BR, Vaillancourt B, Lin H, Massa AN, Geoffroy M, Lundback S, DellaPenna D, Buell CR, Sharma SK, Marshall DF, Waugh R, Bryan GJ, Destefanis M, Nagy I, Milbourne D, Thomson SJ, Fiers M, Jacobs JM, Nielsen KL, Sønderkær M, Iovene M, Torres GA, Jiang J, Veilleux RE, Bachem CW, de Boer J, Borm T, Kloosterman B, van Eck H, Datema E, Hekkert Bt, Goverse A, van Ham RC, and Visser RG
- Subjects
- Evolution, Molecular, Gene Duplication, Gene Expression Regulation, Plant, Genes, Plant genetics, Genetic Variation, Haplotypes genetics, Heterozygote, Homozygote, Immunity, Innate, Inbreeding, Molecular Sequence Annotation, Molecular Sequence Data, Plant Diseases genetics, Ploidies, Solanum tuberosum physiology, Genome, Plant genetics, Genomics, Solanum tuberosum genetics
- Abstract
Potato (Solanum tuberosum L.) is the world's most important non-grain food crop and is central to global food security. It is clonally propagated, highly heterozygous, autotetraploid, and suffers acute inbreeding depression. Here we use a homozygous doubled-monoploid potato clone to sequence and assemble 86% of the 844-megabase genome. We predict 39,031 protein-coding genes and present evidence for at least two genome duplication events indicative of a palaeopolyploid origin. As the first genome sequence of an asterid, the potato genome reveals 2,642 genes specific to this large angiosperm clade. We also sequenced a heterozygous diploid clone and show that gene presence/absence variants and other potentially deleterious mutations occur frequently and are a likely cause of inbreeding depression. Gene family expansion, tissue-specific expression and recruitment of genes to new pathways contributed to the evolution of tuber development. The potato genome sequence provides a platform for genetic improvement of this vital crop., (©2011 Macmillan Publishers Limited. All rights reserved)
- Published
- 2011
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28. The genome of woodland strawberry (Fragaria vesca).
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Shulaev V, Sargent DJ, Crowhurst RN, Mockler TC, Folkerts O, Delcher AL, Jaiswal P, Mockaitis K, Liston A, Mane SP, Burns P, Davis TM, Slovin JP, Bassil N, Hellens RP, Evans C, Harkins T, Kodira C, Desany B, Crasta OR, Jensen RV, Allan AC, Michael TP, Setubal JC, Celton JM, Rees DJ, Williams KP, Holt SH, Ruiz Rojas JJ, Chatterjee M, Liu B, Silva H, Meisel L, Adato A, Filichkin SA, Troggio M, Viola R, Ashman TL, Wang H, Dharmawardhana P, Elser J, Raja R, Priest HD, Bryant DW Jr, Fox SE, Givan SA, Wilhelm LJ, Naithani S, Christoffels A, Salama DY, Carter J, Lopez Girona E, Zdepski A, Wang W, Kerstetter RA, Schwab W, Korban SS, Davik J, Monfort A, Denoyes-Rothan B, Arus P, Mittler R, Flinn B, Aharoni A, Bennetzen JL, Salzberg SL, Dickerman AW, Velasco R, Borodovsky M, Veilleux RE, and Folta KM
- Subjects
- Algorithms, Chloroplasts genetics, Chromosome Mapping, Gene Expression Profiling, Genes, Plant, Genetic Linkage, In Situ Hybridization, Fluorescence, Likelihood Functions, Models, Genetic, Phylogeny, Terminal Repeat Sequences, Transcription, Genetic, Fragaria genetics, Genome, Plant
- Abstract
The woodland strawberry, Fragaria vesca (2n = 2x = 14), is a versatile experimental plant system. This diminutive herbaceous perennial has a small genome (240 Mb), is amenable to genetic transformation and shares substantial sequence identity with the cultivated strawberry (Fragaria × ananassa) and other economically important rosaceous plants. Here we report the draft F. vesca genome, which was sequenced to ×39 coverage using second-generation technology, assembled de novo and then anchored to the genetic linkage map into seven pseudochromosomes. This diploid strawberry sequence lacks the large genome duplications seen in other rosids. Gene prediction modeling identified 34,809 genes, with most being supported by transcriptome mapping. Genes critical to valuable horticultural traits including flavor, nutritional value and flowering time were identified. Macrosyntenic relationships between Fragaria and Prunus predict a hypothetical ancestral Rosaceae genome that had nine chromosomes. New phylogenetic analysis of 154 protein-coding genes suggests that assignment of Populus to Malvidae, rather than Fabidae, is warranted.
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- 2011
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29. Implementing reverse genetics in Rosaceae: analysis of T-DNA flanking sequences of insertional mutant lines in the diploid strawberry, Fragaria vesca.
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Oosumi T, Ruiz-Rojas JJ, Veilleux RE, Dickerman A, and Shulaev V
- Subjects
- Base Sequence, DNA, Bacterial genetics, DNA, Plant genetics, Molecular Sequence Data, Plants, Genetically Modified genetics, Sequence Analysis, DNA, Sequence Homology, Nucleic Acid, Transformation, Genetic, Fragaria genetics, Mutagenesis, Insertional
- Abstract
Reverse genetics is used for functional genomics research in model plants. To establish a model system for the systematic reverse genetics research in the Rosaceae family, we analyzed genomic DNA flanking the T-DNA insertions in 191 transgenic plants of the diploid strawberry, Fragaria vesca. One hundred and seventy-six T-DNA flanking sequences were amplified from the right border (RB) and 37 from the left border (LB) by thermal asymmetric interlaced PCR. Analysis of the T-DNA nick positions revealed that T-DNA was most frequently nicked at the cleavage sites. Analysis of 11 T-DNA integration sites indicated that T-DNA was integrated into the F. vesca genome by illegitimate recombination, as reported in other model plants: Arabidopsis, rice and tobacco. First, deletion of DNA was found at T-DNA integration target sites in all transgenic plants tested. Second, microsimilarities of a few base pairs between the left and/or right ends of the T-DNA and genomic sites were found in all transgenic plants tested. Finally, filler DNA was identified in four break-points. Out of 191 transgenic plants, T-DNA flanking sequences of 79 plants (41%) showed significant similarity to genes, elements or proteins of other plant species and 67 (35%) of the sequences are still unknown strawberry gene fragments. T-DNA flanking sequences of 126 plants (66%) showed homology to plant ESTs. This is the first report of T-DNA integration in a sizeable population of a rosaceous species. We have shown in this paper that T-DNA integration in strawberry is not random but directed by sequence microsimilarities in the host genome.
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- 2010
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30. Development of efficient plant regeneration and transformation system for impatiens using Agrobacterium tumefaciens and multiple bud cultures as explants.
- Author
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Dan Y, Baxter A, Zhang S, Pantazis CJ, and Veilleux RE
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- Blotting, Southern, DNA, Bacterial genetics, Green Fluorescent Proteins genetics, Kanamycin pharmacology, Protein Synthesis Inhibitors pharmacology, Agrobacterium tumefaciens genetics, Impatiens genetics, Impatiens growth & development, Impatiens microbiology, Plants, Genetically Modified, Regeneration genetics, Transformation, Genetic drug effects
- Abstract
Background: Impatiens (Impatiens walleriana) is a top selling floriculture crop. The potential for genetic transformation of Impatiens to introduce novel flower colors or virus resistance has been limited by its general recalcitrance to tissue culture and transformation manipulations. We have established a regeneration and transformation system for Impatiens that provides new alternatives to genetic improvement of this crop., Results: In a first step towards the development of transgenic INSV-resistant Impatiens, we developed an efficient plant regeneration system using hypocotyl segments containing cotyledonary nodes as explants. With this regeneration system, 80% of explants produced an average of 32.3 elongated shoots per initial explant plated, with up to 167 elongated shoots produced per explant. Rooting efficiency was high, and 100% of shoots produced roots within 12 days under optimal conditions, allowing plant regeneration within approximately 8 weeks. Using this regeneration system, we developed an efficient Agrobacterium-mediated Impatiens transformation method using in vitro multiple bud cultures as explants and a binary plasmid (pHB2892) bearing gfp and nptII genes. Transgenic Impatiens plants, with a frequency up to 58.9%, were obtained within 12 to 16 weeks from inoculation to transfer of transgenic plants to soil. Transgenic plants were confirmed by Southern blot, phenotypic assays and T1 segregation analysis. Transgene expression was observed in leaves, stems, roots, flowers, and fruit. The transgenic plants were fertile and phenotypically normal., Conclusion: We report the development of a simple and efficient Agrobacterium-mediated transformation system for Impatiens. To the best of our knowledge, there have been no reports of Agrobacterium-mediated transformation of Impatiens with experimental evidence of stable integration of T-DNA and of Agrobacterium-mediated transformation method for plants using in vitro maintained multiple bud cultures as explants. This transformation system has the advantages of 1) efficient, simple and rapid regeneration and transformation (with no need for sterilization or a greenhouse to grow stock plants), 2) flexibility (available all the time) for in vitro manipulation, 3) uniform and desirable green tissue explants for both nuclear and plastid transformation using Agrobacterium-mediated and biolistics methods, 4) no somaclonal variation and 5) resolution of necrosis of Agrobacterium-inoculated tissues.
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- 2010
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31. SNP discovery and genetic mapping of T-DNA insertional mutants in Fragaria vesca L.
- Author
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Ruiz-Rojas JJ, Sargent DJ, Shulaev V, Dickerman AW, Pattison J, Holt SH, Ciordia A, and Veilleux RE
- Subjects
- Computational Biology, DNA Primers, DNA, Plant genetics, Genetic Linkage, Genotype, Mutagenesis, Insertional, Polymerase Chain Reaction, Chromosome Mapping, Chromosomes, Plant genetics, DNA, Bacterial genetics, Fragaria genetics, Mutation genetics, Polymorphism, Single Nucleotide genetics
- Abstract
As part of a program to develop forward and reverse genetics platforms in the diploid strawberry [Fragaria vesca L.; (2n = 2x = 14)] we have generated insertional mutant lines by T-DNA mutagenesis using pCAMBIA vectors. To characterize the T-DNA insertion sites of a population of 108 unique single copy mutants, we utilized thermal asymmetric interlaced PCR (hiTAIL-PCR) to amplify the flanking region surrounding either the left or right border of the T-DNA. Bioinformatics analysis of flanking sequences revealed little preference for insertion site with regard to G/C content; left borders tended to retain more of the plasmid backbone than right borders. Primers were developed from F. vesca flanking sequences to attempt to amplify products from both parents of the reference F. vesca 815 x F. bucharica 601 mapping population. Polymorphism occurred as: presence/absence of an amplification product for 16 primer pairs and different size products for 12 primer pairs, For 46 mutants, where polymorphism was not found by PCR, the amplification products were sequenced to reveal SNP polymorphism. A cleaved amplified polymorphic sequence/derived cleaved amplified polymorphism sequence (CAPS/dCAPS) strategy was then applied to find restriction endonuclease recognition sites in one of the parental lines to map the SNP position of 74 of the T-DNA insertion lines. BLAST search of flanking regions against GenBank revealed that 46 of 108 flanking sequences were close to presumed strawberry genes related to annotated genes from other plants.
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- 2010
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32. Multiple models for Rosaceae genomics.
- Author
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Shulaev V, Korban SS, Sosinski B, Abbott AG, Aldwinckle HS, Folta KM, Iezzoni A, Main D, Arús P, Dandekar AM, Lewers K, Brown SK, Davis TM, Gardiner SE, Potter D, and Veilleux RE
- Subjects
- Biodiversity, Computational Biology, Fragaria genetics, Humans, Malus genetics, Nutritive Value, Phylogeny, Prunus genetics, Genomics, Rosaceae genetics
- Abstract
The plant family Rosaceae consists of over 100 genera and 3,000 species that include many important fruit, nut, ornamental, and wood crops. Members of this family provide high-value nutritional foods and contribute desirable aesthetic and industrial products. Most rosaceous crops have been enhanced by human intervention through sexual hybridization, asexual propagation, and genetic improvement since ancient times, 4,000 to 5,000 B.C. Modern breeding programs have contributed to the selection and release of numerous cultivars having significant economic impact on the U.S. and world markets. In recent years, the Rosaceae community, both in the United States and internationally, has benefited from newfound organization and collaboration that have hastened progress in developing genetic and genomic resources for representative crops such as apple (Malus spp.), peach (Prunus spp.), and strawberry (Fragaria spp.). These resources, including expressed sequence tags, bacterial artificial chromosome libraries, physical and genetic maps, and molecular markers, combined with genetic transformation protocols and bioinformatics tools, have rendered various rosaceous crops highly amenable to comparative and functional genomics studies. This report serves as a synopsis of the resources and initiatives of the Rosaceae community, recent developments in Rosaceae genomics, and plans to apply newly accumulated knowledge and resources toward breeding and crop improvement.
- Published
- 2008
- Full Text
- View/download PDF
33. Phenotypic and transcriptomic changes associated with potato autopolyploidization.
- Author
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Stupar RM, Bhaskar PB, Yandell BS, Rensink WA, Hart AL, Ouyang S, Veilleux RE, Busse JS, Erhardt RJ, Buell CR, and Jiang J
- Subjects
- Biological Evolution, Gene Expression Profiling, Genome, Plant, Oligonucleotide Array Sequence Analysis, Phenotype, Plant Leaves genetics, Plant Roots genetics, Polyploidy, Solanum tuberosum anatomy & histology, Solanum tuberosum genetics
- Abstract
Polyploidy is remarkably common in the plant kingdom and polyploidization is a major driving force for plant genome evolution. Polyploids may contain genomes from different parental species (allopolyploidy) or include multiple sets of the same genome (autopolyploidy). Genetic and epigenetic changes associated with allopolyploidization have been a major research subject in recent years. However, we know little about the genetic impact imposed by autopolyploidization. We developed a synthetic autopolyploid series in potato (Solanum phureja) that includes one monoploid (1x) clone, two diploid (2x) clones, and one tetraploid (4x) clone. Cell size and organ thickness were positively correlated with the ploidy level. However, the 2x plants were generally the most vigorous and the 1x plants exhibited less vigor compared to the 2x and 4x individuals. We analyzed the transcriptomic variation associated with this autopolyploid series using a potato cDNA microarray containing approximately 9000 genes. Statistically significant expression changes were observed among the ploidies for approximately 10% of the genes in both leaflet and root tip tissues. However, most changes were associated with the monoploid and were within the twofold level. Thus, alteration of ploidy caused subtle expression changes of a substantial percentage of genes in the potato genome. We demonstrated that there are few genes, if any, whose expression is linearly correlated with the ploidy and can be dramatically changed because of ploidy alteration.
- Published
- 2007
- Full Text
- View/download PDF
34. Retrotransposon-based markers from potato monoploids used in somatic hybridization.
- Author
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Lightbourn GJ, Jelesko JG, and Veilleux RE
- Subjects
- Base Sequence, Blotting, Southern, Genetic Markers, Molecular Sequence Data, Polymorphism, Restriction Fragment Length, Sequence Analysis, DNA, Chimera genetics, Hybridization, Genetic, Retroelements genetics, Solanum tuberosum genetics
- Abstract
In an attempt to remove lethal and deleterious genes and enhance the heterozygosity of the potato genome, we developed several diverse somatic hybrids through the electrofusion of selected monoploids. Somatic hybrids and somaclones resulting from fused and unfused protoplasts, respectively, were verified with microsatellites. Molecular markers anchored in the Tst1 retrotransposon were used to examine polymorphisms in the regenerated plants and to reveal any somaclonal variation. Inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon display (sequence-specific amplified polymorphism (S-SAP), anchored in a retransposon) were examined on an ALFexpress DNA sequencer. Because of inconsistencies in the number and quality of bands revealed by the combination of either class of marker in combination with the ALFexpress, we cloned and sequenced 11 S-SAP bands to use as restriction fragment length polymorphism (RFLP) probes in Southern blot analyses of genetic relationships in our potato populations and among related Solanaceae. Readily scorable bands (n = 27) that separated somatic hybrids from parental monoploids and somaclones and grouped monoploids according to known genetic relationships were produced. Some of the probes could be used to differentiate tomato and Datura from potato. Sequence analysis of 5 cloned IRAP and 11 cloned S-SAP markers confirmed that they were anchored in the Tst1 retrotransposon. BLAST searches within GenBank produced 10 highly significant hits (5 nucleotide, 4 expressed sequence tag (EST), and 1 protein) within closely related Solanaceae, suggesting that Tst1 represents an old retroelement that was inserted before the diversion of genera within Solanaceae; however, most sequences were undescribed.
- Published
- 2007
- Full Text
- View/download PDF
35. High-efficiency transformation of the diploid strawberry (Fragaria vesca) for functional genomics.
- Author
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Oosumi T, Gruszewski HA, Blischak LA, Baxter AJ, Wadl PA, Shuman JL, Veilleux RE, and Shulaev V
- Subjects
- Cinnamates pharmacology, Fragaria anatomy & histology, Fragaria growth & development, Green Fluorescent Proteins analysis, Hygromycin B analogs & derivatives, Hygromycin B pharmacology, Mutagenesis, Insertional methods, Plant Shoots genetics, Plant Shoots growth & development, Plants, Genetically Modified anatomy & histology, Plants, Genetically Modified genetics, Rhizobium, Diploidy, Fragaria genetics, Genomics methods, Transformation, Genetic
- Abstract
Fragaria vesca L., a diploid (2n = 2x = 14) relative of the commercial octoploid strawberry, is an attractive model for functional genomics research in Rosaceae. Its small genome size, short reproductive cycle, and facile vegetative and seed propagation make F. vesca a promising candidate for forward and reverse genetics experiments. However, the lack of a high-efficiency transformation protocol required for systematic production of thousands of T-DNA insertional mutant lines and high-throughput gene validation is a major bottleneck. We describe a new transformation procedure that uses leaf explants from newly unfolded trifoliate leaves obtained from stock plants 6-7 weeks after seed germination, co-cultivation with Agrobacterium strain GV3101, and stringent selection on MS medium containing 4 mg l(-1) hygromycin. Using this protocol we achieved 100% transformation efficiency for 6 of 14 F. vesca accessions tested. Accession PI 551572 was determined to be the best candidate for a model in F. vesca functional genomics research, as it showed the greatest propensity for callus formation, transformation, shoot regeneration, ex vitro establishment, and plant growth, requiring only 14-15 weeks to complete its life cycle in different seasons in the greenhouse.
- Published
- 2006
- Full Text
- View/download PDF
36. Microsatellite marker analysis of an anther-derived potato family: skewed segregation and gene-centromere mapping.
- Author
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Chani E, Ashkenazi V, Hillel J, and Veilleux RE
- Subjects
- Alleles, Chromosome Mapping, Chromosome Segregation, Crosses, Genetic, DNA, Plant analysis, DNA, Plant isolation & purification, Diploidy, Genetic Linkage, Genetic Markers, Heterozygote, Inbreeding, Minisatellite Repeats, Ploidies, Polymorphism, Genetic, Centromere genetics, Gene Frequency, Genes, Plant, Microsatellite Repeats, Solanum tuberosum genetics
- Abstract
Segregation patterns of polymorphic simple sequence repeat (SSR) primer pairs were investigated in monoploid potato families derived from anther culture. A total of 14 primers developed from the sequences in the database, as well as from a genomic library of potato, was used. Distorted segregation was observed for seven (50%) polymorphic loci among monoploids derived from an interspecific hybrid. Similar distortion was observed for only one of five loci that could be contrasted between the two monoploid families. Segregation distortion was less common in the sexually derived backcross population between the interspecific hybrid and either of its parents. One locus could be putatively linked to a lethal allele because it showed distorted segregation in both monoploid families, a group of 70 heterozygous diploids derived from unreduced gametes through anther culture, and a backcross population. These diploids were used to map the polymorphic SSR markers with respect to the centromeres using half-tetrad analysis. The majority of the SSR loci mapped more than 33 cM from the centromere, suggesting the occurrence of a single crossover per chromosome arm.
- Published
- 2002
- Full Text
- View/download PDF
37. Development of microsatellite markers in potato and their use in phylogenetic and fingerprinting analyses.
- Author
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Ashkenazi V, Chani E, Lavi U, Levy D, Hillel J, and Veilleux RE
- Subjects
- Base Sequence, DNA Primers, DNA, Plant genetics, Databases, Factual, Hybridization, Genetic, Phylogeny, Polymorphism, Genetic, DNA Fingerprinting, Genetic Markers, Microsatellite Repeats genetics, Solanum tuberosum genetics
- Abstract
Three genomic libraries were constructed using a mixture of DNA from Solanum phureja Juz. & Buk., and S. chacoense Bitt. Two of the libraries were enriched for ATT and GT repeats (a 27-fold enrichment was achieved). In total, 3500 clones of the conventional library, 1,000 of the library enriched for ATT, and 12,000 of the one enriched for GT were screened with five different repeat motifs, and a total of 18 primer pairs was obtained. Another group of 12 primer pairs was obtained from the SSR-containing sequences in the public databases (18 SSR-containing sequences were utilized). From among 30 newly developed primer pairs, 12 previously published ones, and 12 pairs developed for tomato, 7 were used to identify 12 different potato cultivars and introductions, and 12 were used to study phylogenetic distance among seven wild and cultivated potato species. Two SSR markers were sufficient to discriminate the 12 cultivars. The mean number of alleles per polymorphic locus was 5 for the 12 cultivars and 4.5 for the seven species. The results obtained in this study confirm those achieved in similar studies in other plant species regarding the abundance and use of SSR markers in identifying species and cultivars.
- Published
- 2001
- Full Text
- View/download PDF
38. Analysis of the genetic composition of anther-derived potato by randomly amplified polymorphic DNA and simple sequence repeats.
- Author
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Veilleux RE, Shen LY, and Paz MM
- Subjects
- Alleles, Base Sequence, DNA, Plant genetics, Diploidy, Genetic Carrier Screening, Genetic Markers, Haploidy, Molecular Sequence Data, Microsatellite Repeats genetics, Polymorphism, Genetic, Random Amplified Polymorphic DNA Technique, Solanum tuberosum genetics
- Abstract
Randomly amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) analyses were used to characterize the genetic composition of anther-derived plants of a diploid potato clone, CP2 (Solanum chacoense 80-1 x S. phureja 1-3). The ploidy of anther-derived plants was first determined by flow cytometry. A total of 44 decamer primers was screened for RAPD polymorphism. The loci that segregated were selected and scored. The monoploids had less than half as many loci carrying RAPD markers compared with the anther donor. Among 14 anther-derived diploids, 5 were identified as homozygous by marker frequency similar to monoploids and 9 as heterozygous. Five of seven SSRs obtained from published potato sequences were polymorphic in CP2. CP2 was found to be heterozygous with two alleles at four SSR loci (TC/TA, AAG, AGA, CTT) and three alleles at a ACTC locus. Primer pairs flanking each of the five polymorphic SSRs revealed that monoploids had only the allele contributed by S. chacoense 80-1. Homozygous diploids had only one band per SSR locus, whereas heterozygous diploids displayed more than one allele for at least one SSR locus. Results of the SSR analysis supported the findings based on RAPD markers; the same five diploid clones were characterized as homozygous by both SSR and RAPD markers.
- Published
- 1995
- Full Text
- View/download PDF
39. Variation for 2n pollen production in clones of Solanum phureja Juz. and Buk.
- Author
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Veilleux RE and Lauer FI
- Abstract
Frequency of unreduced pollen grains was estimated for five genotypes of Solanum phureja (2n=24) growing in three environments; (E1) cool (7.2-13.3°C) and (E2) warm (12.2-17.2°C) growth chambers and (E3) field conditions. Highly variable frequencies were found, with genotype, environment, and genotype x environment interaction as significant components of variance. The frequency of unreduced gametes for two additional genotypes was studied over time in two growth chamber environments (cool and warm). One genotype, characterized by mostly fused spindles at the second meiotic division, expressed a high frequency of big pollen (BP) in both environments, whereas the second, characterized by fused, parallel and tripolar second division spindless was found to increase in BP frequency over time in the cool chamber, but remained consistently low in the warm chamber. The identification of specific environmental components with general effect on the expression of un-reduced gametes is not possible because of the large genotype x environment interaction component of variance. A genetic hypothesis based on incomplete penetrance and variable expressivity of a dominant gene is presented as an alternative to the currently accepted theory of control of parallel spindles by a single recessive gene.
- Published
- 1981
- Full Text
- View/download PDF
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