Abstract: A segregating doubled haploid (DH) population (n = 96) was developed by anther culture of an F1 plant crossed between susceptible (Manganji) and resistant (Criollo de Morelos 334) lines of pepper (Capsicum annuum L.) to conduct a genetic analysis of resistance to Phytophthora rot caused by Phytophthora capsici. In order to perform a quantitative trait locus (QTL) analysis, we constructed a high density simple sequence repeat (SSR)-based map with a total length of 878 cM. Sixteen linkage groups (LGs) and 118 SSR markers were located using the 626 SSR markers that we previously developed. Resistance was evaluated in two root inoculation tests. Interval mapping for the resistance to P. capsici detected a common major QTL in the duplicate tests and a minor QTL specific to the first test. The major QTL was located on LG15 and flanked with an SSR marker, CAMS420. In addition, seven SSR markers were located within 21 cM intervals from the peak of this QTL. In contrast, the QTL on LG3 was detected with small effects in the first test, the nearest marker was a dominant amplified fragment length polymorphism (AFLP) marker, and the QTL was surrounded by eight SSR markers within a distance of 10 cM. Since some of the linkage markers for agriculturally valuable traits cannot detect polymorphism within breeding populations in C. annuum, the present linkage markers may widen the choice in marker-assisted selection in breeding programs for Phytophthora rot resistant pepper cultivars.
Abstract: An efficient technique for isolation of resistant gene analogs (RGAs) in pepper from silver stained denaturing polyacrylamide gel was developed using a modified amplified fragment length polymorphism (AFLP) strategy. Pepper DNA was digested, ligated and pre-amplified as in a normal AFLP method. The selective amplification was made by using combinations with oligonucleotide primers based on conserved motifs in and around nucleotide binding site (NBS) of known NBS-leucine-rich repeats resistance proteins from known resistant genes. The amplified products were separated by using denaturing polyacrylamide gels and silver staining instead of radioactive labelling. We isolated specific polymorphic AFLP bands directly from the gels with one round of polymerase chain reaction amplification, in order to confirm, after sequencing, that these bands have homologies with products of resistance genes described so far. Two bands (R2: 250 bp and R6: 150 bp) are particularly highlighted because they could be considered as RGAs related to resistance to Phytophthora capsici in pepper, because their sequences have a very high homology with other resistant gene analogs that have already been described. Besides, they were only detected in the resistant parent and in the bulked resistant segregants but not in the susceptible parent or susceptible F2 segregants. We can conclude that the technique used is clean, quick and efficient for the isolation of RGAs in pepper.
Abstract: Quantitative trait loci (QTL) mapping for agronomic traits was carried out in cocoa (Theobroma cacao L.). Regions of the genome involved in yield, vigor, and resistance to Phytophthora palmivora were identified. Three
heterozygous clones, one upper Amazon Forastero (IMC78) and two Trinitario (DR1 and S52), were crossed with the same male parent, a lower Amazon Forastero (Catongo), known to be highly homozygous. Observations were made on progeny over nine consecutive years. One to three QTL related to yield were detected in each of the three populations, located on chromosomes 1, 2, 4, 5, 9, and 10. They explained between 8.1 and 19.3% of the phenotypic variation and showed various levels of repeatability. In IMC78, the QTL detected on chromosome 5 was the most repeatable over years. The QTL for the average individual pod weight on chromosome 4 was the most significant with an LOD of 17.3 and an R2 of 43.7. QTL related to these traits were identified in the same region of the genome in clones of different genetic groups. This suggests that molecular markers can be used to improve cocoa varieties.
Abstract: The pepper accession Criollo de Morelos 334 is the most efficient source of resistance currently known to Phytophthora capsici and P. parasitica. To investigate whether genetic controls of resistance to two Phytophthora species are independent, we compared the genetic architecture of resistance of CM334 to both Phytophthora species. The RIL population F5YC used to construct the high-resolution genetic linkage map of pepper was assessed for resistance to one isolate of each Phytophthora species. Inheritance of the P. capsici and P. parasitica resistance was polygenic. Twelve additive QTLs involved in the P. capsici resistance and 14 additive QTLs involved in the P. parasitica resistance were detected. The QTLs identified in this progeny were specific to these Phytophthora species. Comparative mapping analysis with literature data identified three colocations between resistance QTLs to P. parasitica and P. capsici in pepper. Whereas this result suggests presence of common resistance factors to the two Phytophthora species in pepper, which possibly derive from common ancestral genes, calculation of the colocation probability indicates that these colocations could occur by chance.
Abstract:Piper colubrinum Link., the exotic wild Piper shows high degree of resistance to fungal pathogens and is a potential source of resistance genes. A PCR-based suppression subtractive hybridization (SSH) was used to identify P. colubrinum genes that are differentially expressed in response to the signalling molecule, salicylic acid(SA). A subtracted library of SA-induced genes was synthesized and one of the clones showed sequence homology to osmotin, a member of class-V group of pathogenesis-related (PR) gene family. The 315 bp gene fragment was used to probe total RNA prepared from untreated and SA-treated leaf tissues. Osmotin fragment cloned from the subtracted library was also used to probe RNA from ethylene-treated leaf tissues. Northern blot analysis revealed that osmotin is dominantly expressed in SA/ethylene-treated tissue. This indicates that SSH can be used to identify and clone PR genes in P. colubrinum.
Abstract: Late blight caused by the oomycete Phytophthora infestans is the most important fungal disease in potato cultivation worldwide. Resistance to late blight is controlled by a few major genes (R genes) which can be easily overcome by new races of P. infestans and/or by an unknown number of genes expressing a quantitative type of resistance which may be more durable. Quantitative resistance of foliage to late blight was evaluated in five F1 hybrid families originating from crosses among seven different diploid potato clones. Tuber resistance was evaluated in four of the families. Two of the families were scored for both foliage maturity and vigour. The five families were genotyped with DNA-based markers and tested for linkage with the traits analysed. QTL (quantitative trait locus) analysis identified at least twelve segments on ten chromosomes of potato having genes that affect reproducibly foliage resistance. Two of those segments also have major R genes for resistance to late blight. The segments are tagged by 21 markers that can be analyzed based on PCR (polymerase chain reaction) with specific oligonucleotide primers. One QTL was detected for tuber resistance and one for foliage vigour. Two QTLs were mapped for foliage maturity. Major QTL effects on foliage and tuber resistance to late blight and on foliage maturity and vigour were all linked with marker GP179 on linkage group V of potato. Plants having alleles at this QTL, which increased foliage resistance, exhibited decreased tuber resistance, later maturity and more vigour.
Abstract: A doubled-haploid (DH) population (n = 176) obtained by anther culture of an F1 hybrid between a line susceptible to Phytophthora capsici K9-11 (Capsicum annuum L.) and a line resistant to P. capsici AC2258 (C. annuum L.) was inoculated with P. capsici. QTL analysis of the resistance was performed using a linkage map consisting of 16 linkage groups (LGs), covering a total distance of 1100.5 cM. Three QTLs were detected on LG1, LG6 and LG7. The QTL with the highest LOD score, detected on LG7, explained 82.7% of the phenotypic variance with a LOD score of 67.02. This QTL was designated as Phyt-1. The nearest marker was an AFLP marker, M10E3-6. The second QTL, designated as Phyt-2, was found on LG1. It explained 6.4% of the phenotypic variance with a LOD score of 2.54. The nearest RAPD marker was RP13-1. The other QTL, designated as Phyt-3, which was found on LG6, explained 5.6% of the phenotypic variance with a LOD score of 2.20. The nearest AFLP marker was M9E3-11. It was confirmed that the lines with a high resistance could be efficiently selected by using two markers, M10E3-6 and RP13-1, simultaneously. The presence of both Phyt-1 and Phyt-2 under homozygous conditions may enable to breed resistant cultivars of sweet pepper. The molecular markers identified in the present study could be useful for marker-assisted selection (MAS) in order to breed sweet pepper cultivars with a high resistance to P. capsici using AC2258 as a source of resistance genes.
Abstract: This study aimed to compare the genetic control of cacao resistance to three species of Phytophthora: Phytophthora palmivora, Phytophthora megakarya and Phytophthora capsici. The study was conducted on 151 hybrid progenies created in Cote d Ivoire and grown in a green-house in Montpellier. Phytophthora resistance was screened by leaf-test inoculation with two different strains per species. Selection of the best individuals for resistance to P. palmivora at a 10% selection rate, would lead to a genetic progress of 47% in the disease evaluation for this species and a genetic progress of 42% and 21% for the two other species. A genetic map with a total length of 682 cM was built with 213 markers, 190 AFLPs and 23 microsatellites. QTLs were identified using composite interval mapping. QTLs were found located in six genomic regions. One of these was detected with five strains belonging to the three Phytophthora species. Two other regions were detected with two or three strains of two different species. Three additional QTLs were detected for only one species of Phytophthora. Each QTL explained between 8 to 12% of the phenotypic variation. For each strain, between 11.5% to 27.5% of the total phenotypic variation could be explained by the QTLs identified. The identification of multiple QTLs involved in resistance to Phytophthora offers the possibility to improve durability of resistance in cocoa by a possible cumulation of many different resistance genes located in different chromosome regions using marker-aided selection.
Abstract: Tomato bacterial wilt caused by Ralstonia solanacearum is a model system for studying plant-bacterial interactions, because it is genetically one of the best characterized plant diseases. We demonstrate here that four different strains of R. solanacearum, two from radishes (Rd4 and Rd15) and two from tomato (Ps21 and Ps95), can infect 27 different ecotypes of Arabidopsis thaliana, causing different responses. All ecotypes tested were highly susceptible to strain Rd15, which caused symptoms similar to those observed in tomato plants. For example, leaf drooping and discoloration developed just 3 days after inoculation, and plants completely wilted within 1 week. Strains Rd4 and Ps95 were less infectious than Rd15. With these two strains, a variety of disease responses were observed among different ecotypes at 2 weeks after inoculation; both susceptible and resistant ecotypes of A. thaliana were identified. Ps21 was the least infectious of the four strains and caused almost no symptoms in any of the ecotypes of Arabidopsis tested. Direct bacterial isolation and plant skeleton hybridization analysis from infected plants indicated that bacterial colonization was correlated with the severity of symptoms. Growth of bacteria was limited to the infection site in resistant plants, whereas the bacteria spread throughout susceptible plants by 1 week after inoculation.
Abstract: Bacterial wilt incidence was reduced by 38.1% and 100% in silicon-treated plants of the moderately resistant tomato genotype King Kong 2 and the resistant genotype Hawaii 7998 grown in peat substrate. At 5 days post inoculation the bacterial population was significantly reduced in stems and roots of genotype Hawaii 7998, and in stems of King Kong 2 in silicon-treated plants compared to non-treated plants, indicating a silicon-induced resistance, since silicon accumulated in roots, but not in stems, while a tolerance effect was observed in the susceptible genotype L390. Characterization of possible molecular mechanisms involved in silicon-mediated resistance by immuno-histochemical analysis of stem cell walls indicated silicon-induced changes in the pectic polysaccharide structure. After infection homogalacturonan with non-blockwise degradation of methyl-esters was increased in vessel walls in non-silicon-treated plants, but not in silicon-treated plants, possibly indicating the action of pathogen pectinmethylesterase. Also the staining of vessel walls for arabinogalactan-protein in infected, non-silicon-treated plants was not observed in silicon-treated plants. In inoculated, silicon-treated plants, staining for arabinan side chains of rhamnogalacturonan I (RG I) was increased in some vessel walls, and fluorescence of antibodies for galactan side chains of RG I overall increased in the xylem parenchyma compared to non-silicon-amended plants. These observations suggest an induced basal resistance on cell wall level after silicon treatment, while the yellow or brown autofluorescence occurring in inoculated, non-silicon-treated plants disappeared.
Abstract