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22 result(s) for "Keulemans, Johan"
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Regulation of fruit ascorbic acid concentrations during ripening in high and low vitamin C tomato cultivars
Background To gain insight into the regulation of fruit ascorbic acid (AsA) pool in tomatoes, a combination of metabolite analyses, non-labelled and radiolabelled substrate feeding experiments, enzyme activity measurements and gene expression studies were carried out in fruits of the ‘low-’ and ‘high-AsA’ tomato cultivars ‘Ailsa Craig’ and ‘Santorini’ respectively. Results The two cultivars exhibited different profiles of total AsA (totAsA, AsA + dehydroascorbate) and AsA accumulation during ripening, but both displayed a characteristic peak in concentrations at the breaker stage. Substrate feeding experiments demonstrated that the L-galactose pathway is the main AsA biosynthetic route in tomato fruits, but that substrates from alternative pathways can increase the AsA pool at specific developmental stages. In addition, we show that young fruits display a higher AsA biosynthetic capacity than mature ones, but this does not lead to higher AsA concentrations due to either enhanced rates of AsA breakdown (‘Ailsa Craig’) or decreased rates of AsA recycling (‘Santorini’), depending on the cultivar. In the later stages of ripening, differences in fruit totAsA-AsA concentrations of the two cultivars can be explained by differences in the rate of AsA recycling activities. Analysis of the expression of AsA metabolic genes showed that only the expression of one orthologue of GDP-L-galactose phosphorylase ( SlGGP1 ), and of two monodehydroascorbate reductases ( SlMDHAR1 and SlMDHAR3 ) correlated with the changes in fruit totAsA-AsA concentrations during fruit ripening in ‘Ailsa Craig’, and that only the expression of SlGGP1 was linked to the high AsA concentrations found in red ripe ‘Santorini’ fruits. Conclusions Results indicate that ‘Ailsa Craig’ and ‘Santorini’ use complementary mechanisms to maintain the fruit AsA pool. In the low-AsA cultivar (‘Ailsa Craig’), alternative routes of AsA biosynthesis may supplement biosynthesis via L-galactose, while in the high-AsA cultivar (‘Santorini’), enhanced AsA recycling activities appear to be responsible for AsA accumulation in the later stages of ripening. Gene expression studies indicate that expression of SlGGP1 and two orthologues of SlMDHAR are closely correlated with totAsA-AsA concentrations during ripening and are potentially good candidates for marker development for breeding and selection.
Allelic Variation in Paralogs of GDP-L-Galactose Phosphorylase Is a Major Determinant of Vitamin C Concentrations in Apple Fruit
To identify the genetic factors underlying the regulation of fruit vitamin C (L-ascorbic acid [AsA]) concentrations, quantitative trait loci (QTL) studies were carried out in an Fl progeny derived from a cross between the apple (Malus X domestica) cultivars Telamón and Braeburn over three years. QTL were identified for AsA, glutathione, total antioxidant activity in both flesh and skin tissues, and various quality traits, including flesh browning. Four regions on chromosomes 10, 11, 16, and 17 contained stable fruit AsA-QTL clusters. Mapping of AsA metabolic genes identified colocations between orthologs of GDP-L-galactose phosphorylase (GGP), dehydroascorbate reducíase (DHAR), and nucleobase-ascorbate transporter within these QTL clusters. Of particular interest are the three paralogs of MdGGP, which all colocated within AsA-QTL clusters. Allelic variants of MdGGPl and MdGGP3 derived from the cultivar Braeburn parent were also consistently associated with higher fruit total AsA concentrations both within the mapping population (up to 10-fold) and across a range of commercial apple germplasm (up to 6-fold). Striking differences in the expression of the cv Braeburn MdGGPl alíele between fruit from high-and low-AsA genotypes clearly indicate a key role for MdGGPl in the regulation of fruit AsA concentrations, and this MdGGP allele-specific single-nucleotide polymorphism marker represents an excellent candidate for directed breeding for enhanced fruit AsA concentrations. Interestingly, colocations were also found between MdDHAR3-3 and a stable QTL for browning in the cv Telamón parent, highlighting links between the redox status of the AsA pool and susceptibility to flesh browning.
Identification and stability of QTLs for fruit quality traits in apple
Breeding for fruit quality traits is complex due to the polygenic (quantitative) nature of the genetic control of these traits. Therefore, to improve the speed and efficiency of genotype selection, attention in recent years has focused on the identification of quantitative trait loci (QTLs) and molecular markers associated with these QTLs. However, despite the huge potential of molecular markers in breeding programmes, their implementation in practice has been limited by the lack of information on the stability of QTLs across different environments and within different genetic backgrounds. Here, we present the results from a comprehensive analysis of the inheritance of fruit quality traits within a population derived from a cross between the apple cultivars 'Telamon' and 'Braeburn' over two successive seasons. A total of 74 different QTLs were identified for all the major fruit physiological traits including fruit height, diameter, weight and stiffness, flesh firmness, rate of flesh browning, acidity, the [ordinal indicator, masculine]Brix content and harvest date. Seventeen of these QTLs were 'major' QTLs, accounting for over 20% of the observed population variance of the trait. However, only one third (26) of the identified QTLs were stable over both harvest years, and of these year-stable QTLs only one was a major QTL. A direct comparison with published QTL results obtained using other populations (King et al., Theor Appl Genet 102:1227-1235, 2001; Liebhard et al., Plant Mol Biol 52:511-526, 2003) is difficult because the linkage maps do not share a sufficient number of common markers and due to differences in the trait evaluation protocols. Nonetheless, our results suggest that for the six fruit quality traits which were measured in all populations, nine out of a total of 45 QTLs were common or stable across all population x environments combinations. These results are discussed in the framework of the development and application of molecular markers for fruit quality trait improvement.
Genetic Control of Fruit Vitamin C Contents
An F₁ progeny derived from a cross between the apple (Malus x domestica) cultivars Telamon and Braeburn was used to identify quantitative trait loci (QTL) linked to the vitamin C (L-ascorbate [L-AA]) contents of fruit skin and flesh (cortex) tissues. We identified up to three highly significant QTLs for both the mean L-AA and the mean total L-AA contents of fruit flesh on both parental genetic linkage maps, confirming the quantitative nature of these traits. These QTLs account for up to a maximum of 60% of the total population variation observed in the progeny, and with a maximal individual contribution of 31% per QTL. QTLs common to both parents were identified on linkage groups (LGs) 6, 10, and 11 of the Malus reference map, while each parent also had additional unique QTLs on other LGs. Interestingly, one strong QTL on LG-17 of the Telamon linkage map colocalized with a highly significant QTL associated with flesh browning, and a minor QTL for dehydroascorbate content, supporting earlier work that links fruit L-AA contents with the susceptibility of hardfruit to postharvest browning. We also found significant minor QTLs for skin L-AA and total L-AA (L-AA + dehydroascorbate) contents in Telamon. Currently, little is known about the genetic determinants underlying tissue L-AA homeostasis, but the presence of major, highly significant QTL in both these apple genotypes under field conditions suggests the existence of common control mechanisms, allelic heterozygosity, and helps outline strategies and the potential for the molecular breeding of these traits.
Fine genetic mapping of the Co locus controlling columnar growth habit in apple
Tree architecture is an important, complex and dynamic trait affected by diverse genetic, ontogenetic and environmental factors. ‘Wijcik McIntosh’, a columnar (reduced branching) sport of ‘McIntosh’ and a valuable genetic resource, has been used intensively in apple-breeding programs for genetic improvement of tree architecture. The columnar growth habit is primarily controlled by the dominant allele of gene Co (columnar) on linkage group-10. But the Co locus is not well mapped and the Co gene remains unknown. To precisely map the Co locus and to identify candidate genes of Co, a sequence-based approach using both peach and apple genomes was used to develop new markers linked more tightly to Co. Five new simple sequence repeats markers were developed (C1753-3520, C18470-25831, C6536-31519, C7223-38004 and C7629-22009). The first four markers were obtained from apple genomic sequences on chromosome-10, whereas the last (C7629-22009) was from an unanchored apple contig that contains an apple expressed sequence tag CV082943, which was identified through synteny analysis between the peach and apple genomes. Genetic mapping of these five markers in four F1 populations of 528 genotypes and 290 diverse columnar selections/cultivars (818 genotypes in total) delimited the Co locus in a genetic interval with 0.37 % recombination between markers C1753-3520 and C7629-22009. Marker C18470-25831 co-segregates with Co in the 818 genotypes studied. The Co region is estimated to be 193 kb and contains 26 predicted gene in the ‘Golden Delicious’ genome. Among the 26 genes, three are putative LATERAL ORGAN BOUNDARIES (LOB) DOMAIN (LBD) containing transcription factor genes known of essential roles in plant lateral organ development, and are therefore considered as strong candidates of Co, designated MdLBD1, MdLBD2, and MdLBD3. Although more comprehensive studies are required to confirm the function of MdLBD1-3, the present work represents an important step forward to better understand the genetic and molecular control of tree architecture in apple.
Effect of polyploidization on morphology in two apple (Malus × domestica) genotypes
Because polyploidy often results in enhancement of desirable properties, artificial genome doubling is commonly used in agri- and horticultural crop breeding programs. In this study genome doubling was induced in two apple genotypes. The effect on vegetative morphological and physiological traits of the plants was then comprehensively determined by comparing the obtained tetraploid apple plants with their diploid counterparts. Out of 17 different physio- and morphological characteristics, 15 were significantly affected in one or both genotypes. The response of these 15 characteristics also appeared to have been caused by two effects; 10 of the 15 characteristics exhibited a common response to ploidy change over both genotypes while five traits showed a genotype-specific response to polyploidization. Tetraploid leaves also exhibited a darker leaf colour, which could be correlated to a higher pigment concentration. Furthermore, the results also show a decreased elongation rate and leaf size in tetraploids, which is suggested to be due to the observed lower cell density in the polyploid apple plants.
Heterologous oligonucleotide microarrays for transcriptomics in a non-model species; a proof-of-concept study of drought stress in Musa
Background 'Systems-wide' approaches such as microarray RNA-profiling are ideally suited to the study of the complex overlapping responses of plants to biotic and abiotic stresses. However, commercial microarrays are only available for a limited number of plant species and development costs are so substantial as to be prohibitive for most research groups. Here we evaluate the use of cross-hybridisation to Affymetrix oligonucleotide GeneChip ® microarrays to profile the response of the banana ( Musa spp.) leaf transcriptome to drought stress using a genomic DNA (gDNA)-based probe-selection strategy to improve the efficiency of detection of differentially expressed Musa transcripts. Results Following cross-hybridisation of Musa gDNA to the Rice GeneChip ® Genome Array, ~33,700 gene-specific probe-sets had a sufficiently high degree of homology to be retained for transcriptomic analyses. In a proof-of-concept approach, pooled RNA representing a single biological replicate of control and drought stressed leaves of the Musa cultivar 'Cachaco' were hybridised to the Affymetrix Rice Genome Array. A total of 2,910 Musa gene homologues with a >2-fold difference in expression levels were subsequently identified. These drought-responsive transcripts included many functional classes associated with plant biotic and abiotic stress responses, as well as a range of regulatory genes known to be involved in coordinating abiotic stress responses. This latter group included members of the ERF, DREB, MYB, bZIP and bHLH transcription factor families. Fifty-two of these drought-sensitive Musa transcripts were homologous to genes underlying QTLs for drought and cold tolerance in rice, including in 2 instances QTLs associated with a single underlying gene. The list of drought-responsive transcripts also included genes identified in publicly-available comparative transcriptomics experiments. Conclusion Our results demonstrate that despite the general paucity of nucleotide sequence data in Musa and only distant phylogenetic relations to rice, gDNA probe-based cross-hybridisation to the Rice GeneChip ® is a highly promising strategy to study complex biological responses and illustrates the potential of such strategies for gene discovery in non-model species.
Hyperspectral Reflectance and Fluorescence Imaging to Detect Scab Induced Stress in Apple Leaves
Apple scab causes significant losses in the production of this fruit. A timely and more site-specific monitoring and spraying of the disease could reduce the number of applications of fungicides in the fruit industry. The aim of this leaf-scale study therefore lies in the early detection of apple scab infections in a non-invasive and non-destructive way. In order to attain this objective, fluorescence- and hyperspectral imaging techniques were used. An experiment was conducted under controlled environmental conditions, linking hyperspectral reflectance and fluorescence imaging measurements to scab infection symptoms in a susceptible apple cultivar (Malus x domestica Borkh. cv. Braeburn). Plant stress was induced by inoculation of the apple plants with scab spores. The quantum efficiency of Photosystem II (PSII) photochemistry was derived from fluorescence images of leaves under light adapted conditions. Leaves inoculated with scab spores were expected to have lower PSII quantum efficiency than control (mock) leaves. However, besides scab-induced, also immature leaves exhibited low PSII quantum efficiency. Therefore, this study recommends the simultaneous use of fluorescence imaging and hyperspectral techniques. A shortwave infrared narrow-waveband ratio index (R1480/R2135) is presented in this paper as a promising tool to identify scab stress before symptoms become visible to the naked eye. Low PSII quantum efficiency attended by low narrow waveband R1480/R2135 index values points out scab stress in an early stage. Apparent high PSII quantum efficiency together with high overall reflectance in VIS and SWIR spectral domains indicate a severe, well-developed scab infection.
Effect of polyploidisation on the response of apple (Malus × domestica Borkh.) to Venturia inaequalis infection
Development of new and durable strategies to improve the apple plant’s resistance against apple scab (Venturia inaequalis) remains a major challenge. Polyploids or organisms with three or more complete chromosome sets often possess properties superior to their diploid counterparts. Studies reported that polyploidy confers an increased resistance to biotic and abiotic stress factors. However, the potential use of polyploidy to increase disease resistance is still insufficiently investigated. We determined the influence of artificial genome doubling on the response of three Malus × domestica genotypes, with a variable level of susceptibility to apple scab. Based on visual symptom evaluation and real-time PCR quantification of the V. inaequalis DNA in apple leaves, an increased resistance was observed in the neotetraploid form of the monogenic resistant genotype compared to its diploid progenitor. No pronounced effects were observed comparing ploidy levels of the susceptible genotypes. These results suggest a potential role for polyploidisation in apple scab resistance, but it seems to depend on the degree of disease susceptibility of the genotype. Still, further research on the effects of polyploidy might offer potential in the development of new and durable resistance strategies.
Genetic diversity, population structure, and linkage disequilibrium of elite and local apple accessions from Belgium using the IRSC array
The identification of molecular markers associated with economic and quality traits will help improve breeding for new apple ( Malus × domestica Borkh.) cultivars. Tools such as the 8K apple SNP array developed by the RosBREED consortium allow for high-throughput genotyping of SNP polymorphisms within collections. However, genetic characterization and the identification of population stratification and kinship within germplasm collections is a fundamental prerequisite for identifying robust marker–trait associations. In this study, a collection of apple germplasm originally developed for plant architectural studies and consisting of both non-commercial/local and elite accessions was genotyped using the 8K apple SNP array to identify cryptic relationships between accessions, to analyze population structure and to calculate the linkage disequilibrium (LD). A total of nine pairs of synonyms and several triploids accessions were identified within the 130 accessions genotyped. In addition, most of the known parent-child relations were confirmed, and several putative, previously unknown parent-child relations were identified among the local accessions. No clear subgroups could be identified although some separation between local and elite accessions was evident. The study of LD showed a rapid decay in our collection, indicating that a larger number of SNPs is necessary to perform whole genome association mapping. Finally, an association mapping effort for architectural traits was carried out on a small number of accessions to estimate the feasibility of this approach.