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Papers by theo hendriks

Theoretical and Applied Genetics, May 21, 2013

High-density genetic maps were constructed for loci involved in nuclear male sterility (NMS1-locu... more High-density genetic maps were constructed for loci involved in nuclear male sterility (NMS1-locus) and sporophytic self-incompatibility (S-locus) in chicory (Cichorium intybus L.). The mapping population consisted of 389 F1 0 individuals derived from a cross between two plants, K28 (male-sterile) and K59 (pollen-fertile), both heterozygous at the S-locus. This F1 0 mapping population segregated for both male sterility (MS) and strong selfincompatibility (SI) phenotypes. Phenotyping F1 0 individuals for MS allowed us to map the NMS1-locus to linkage group (LG) 5, while controlled diallel and factorial crosses to identify compatible/incompatible phenotypes mapped the S-locus to LG2. To increase the density of markers around these loci, bulked segregant analysis was used. Bulks and parental plants K28 and K59 were screened using amplified fragment length polymorphism (AFLP) analysis, with a complete set of 256 primer combinations of EcoRI-ANN and MseI-CNN. A total of 31,000 fragments were generated, of which 2,350 showed polymorphism between K59 and K28. Thirteen AFLP markers were identified close to the NMS1-locus and six in the vicinity of the S-locus. From these AFLP markers, eight were transformed into sequence-characterized amplified region (SCAR) markers and of these five showed co-dominant polymorphism. The chromosomal regions containing the NMS1-locus and the S-locus were each confined to a region of 0.8 cM. In addition, we mapped genes encoding proteins similar to S-receptor kinase, the female determinant of sporophytic SI in the Brasicaceae, and also markers in the vicinity of the putative S-locus of sunflower, but none of these genes or markers mapped close to the chicory S-locus.

Copyright © 2012 Meriem Bahri et al. This is an open access article distributed under the Creativ... more Copyright © 2012 Meriem Bahri et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. A “novel ” protocol is presented for easy and reliable estimation of soluble hydroxycinnamate levels in Cichorium intybus L. leaf tissue in large-scale experiments. Samples were standardized by punching 6 discs per leaf, and hydroxycinnamates were extracted by submerging the discs in 80 % ethanol with 5 % acetic acid for at least 48 h in the darkness at 4◦C. Residual dry mass of the discs was used for a posteriori correction of compound levels. Chlorophyll was eliminated by chloroform, and the aqueous phases were transferred to microplates, dried, and dissolved in 50 % methanol for HPLC analysis and storage. An HPLC program of 8min was developed for the analysis of the extracts. Comparisons with extractions of liquid nitrogen powders i...

Background: In our laboratory we use cultured chicory (Cichorium intybus) explants as a model to ... more Background: In our laboratory we use cultured chicory (Cichorium intybus) explants as a model to investigate cell reactivation and somatic embryogenesis and have produced 2 chicory genotypes (K59, C15) sharing a similar genetic background. K59 is a responsive genotype (embryogenic) capable of undergoing complete cell reactivation i.e. cell deand re-differentiation leading to somatic embryogenesis (SE), whereas C15 is a non-responsive genotype (nonembryogenic) and is unable to undergo SE. Previous studies [1] showed that the use of the β-D-glucosyl Yariv reagent (β-GlcY) that specifically binds arabinogalactan-proteins (AGPs) blocked somatic embryo production in chicory root explants. This observation indicates that β-GlcY is a useful tool for investigating somatic embryogenesis (SE) in chicory. In addition, a putative AGP (DT212818) encoding gene was previously found to be significantly up-regulated in the embryogenic K59 chicory genotype as compared to the non-embryogenic C15 genot...

Theoretical and Applied Genetics

Progress in Plant Growth Regulation, 1992

In all plant species investigated it is possible to induce somatic cells of most plant organs to ... more In all plant species investigated it is possible to induce somatic cells of most plant organs to proliferate in liquid synthetic media supplemented with auxins alone or with auxins and cytokinins. Initially in carrot, and later in many other plant species, it was shown that some of these proliferating cells are capable of producing somatic embryos able to develop into fertile flowering plants. The strategy most commonly employed is to expose expiant tissue to a high concentration of a stable and persistent auxin such as 2,4-D. After a period of proliferation of the released non-embryogenic expiant cells in the presence of 2,4-D, a subpopulation of single cells and clusters of cells can be distinguished that have acquired embryogenic potential. From these single embryogenic cells (Komamine et al. 1990) or from clusters of embryogenic cells designated proembryogenic masses (Halperin, 1966), somatic embryos can develop after simple culture manipulations. These usually involve enrichment for proembryogenic masses, followed by a 10 to 100 fold reduction in cell density together with a decrease in the concentration or the complete removal of exogenous auxins. Most embryogenic carrot cultures are maintained over many subcultures with 2,4-D either as the sole growth regulator, or in the presence of both 2,4-D and cytokinin. Therefore, cells that have embryogenic potential may either be continuously formed from non-embryogenic cells, or constitute an independent self-propagating subpopulation. In carrot cultures, depletion of the population of embryogenic cells eventually leads to loss of the embryogenic potential. In other culture systems such as alfalfa, non-embryogenic cells can be maintained in media containing NAA, while a short exposure to a high concentration of 2,4-D is sufficient for these cells to acquire embryogenic potential (Bogre et al. 1990). Although many variations in the source of the original explant material and in culture conditions have been reported, the basic scheme always involves an induction phase in the presence of 2,4-D, during which proliferating cells become embryogenic, and an expression phase in the absence of 2,4-D, during which the actual somatic embryos develop.

The Scientific World Journal, 2012

A “novel” protocol is presented for easy and reliable estimation of soluble hydroxycinnamate leve... more A “novel” protocol is presented for easy and reliable estimation of soluble hydroxycinnamate levels inCichorium intybusL. leaf tissue in large-scale experiments. Samples were standardized by punching 6 discs per leaf, and hydroxycinnamates were extracted by submerging the discs in 80% ethanol with 5% acetic acid for at least 48 h in the darkness at 4°C. Residual dry mass of the discs was used fora posterioricorrection of compound levels. Chlorophyll was eliminated by chloroform, and the aqueous phases were transferred to microplates, dried, and dissolved in 50% methanol for HPLC analysis and storage. An HPLC program of 8 min was developed for the analysis of the extracts. Comparisons with extractions of liquid nitrogen powders indicated that the novel extraction method was reliable. No degradation of the major hydroxycinnamates—caftaric, chlorogenic, and chicoric acids—was observed, during maceration at ambient temperatures, or after storage for 1 year.

Biotechnology Applications of Microinjection, Microscopic Imaging, and Fluorescence, 1993

In cell walls of phellem of potato tubers (Solanum tuberosum cv Bintje) a yellow autofluorescence... more In cell walls of phellem of potato tubers (Solanum tuberosum cv Bintje) a yellow autofluorescence was observed when viewed with blue light (470–490 nm). A similar autofluorescence was also observed in tuber tracheary elements, suggesting that the fluorescence is caused by lignin or lignin-like material. Confocal scanning laser (488 nm) microscopy revealed that in phellem the autofluorescent material is present mainly in the intercellular spaces.

Plant Molecular Biology, 2000

Three different ß-1,3-glucanase cDNA fragments, CG1, CG2 and CG3, were obtained by RT-PCR from RN... more Three different ß-1,3-glucanase cDNA fragments, CG1, CG2 and CG3, were obtained by RT-PCR from RNA isolated from Cichorium hybrid `474' leaf fragments cultured for 11 days under somatic embryogenesis-inducing conditions. When expressed in Escherichia coli the proteins encoded by the three cDNAs were recognized by antibodies raised against 38 kDa extracellular ß-1,3-glucanases studied previously (Helleboid et al., Planta 205 (1998) 56–63). The CG2

Theoretical and Applied Genetics, 1984

In root and flower corolla tissue of Petunia several anodic moving peroxidase isoenzymes are pres... more In root and flower corolla tissue of Petunia several anodic moving peroxidase isoenzymes are present, which cannot be detected in other organs. Alleles of the gene prxF control the presence or absence of several peroxidases that are only present in flower corolla tissue. Alleles of the gene prxG code for two peroxidases that can only be detected in root tissue. In addition to mutations of prxG that cause a change in the electrophoretic mobility of the PRXg enzymes, a mutation was also found that causes the absence of expression in enzyme activity. Crossing experiments indicated that this mutation is located in the gene prxG. Peroxidases encoded by the gene prxH were only found in root tissue. Two alleles of prxH were identified by electrophoretic variation; one allele is responsible for a single band, whereas the other allele could be recognized by a double-banded phenotype. The double-banded PRXh phenotype is suggested to be caused by tandem duplication, followed by mutation in one of the genes. A third prxH allele could be identified by the absence of PRXh activity. The genes prxF, prxG, and prxH were shown to be located on chromosome VII, with the following gene order: prxG-A n4-lap B-gpi B-prx H-prx F.

Theoretical and Applied Genetics, 2013

European Journal of Biochemistry, 1991

The fast-moving anionic peroxidase isoenzyme variant PRXa was purified from leaves of petunia (Pe... more The fast-moving anionic peroxidase isoenzyme variant PRXa was purified from leaves of petunia (Petunia hybrida). Over 1300-fold purification was achieved by subjecting extracellular extracts to two sequential acetone precipitations and resuspending the pellets at pH 5.0 and pH 8.0, respectively, followed by gel filtration and chromatofocusing. The purified enzyme had an absorbance ratio (A405 nm/A280 nm) of 3.6, a molecular mass of about 37 kDa and a pI of 3.8. Three molecular forms with slightly different molecular masses were separated by concanavalin-A--Sepharose affinity chromatography, indicating that these three forms differ in their carbohydrate moieties. The absorption spectrum of PRXa had maxima at 496 and 636 nm and a Soret band at 405 nm. Spectra of compounds I and IV were obtained by titrating a batch of PRXa stored for several months at -20 degrees C with H2O2. The addition of 1 mol H2O2/mol freshly purified PRXa caused the formation of compound II, indicating that freshly isolated PRXa contains a bound hydrogen donor which is lost upon storage. Compound III was obtained from both preparations in the presence of excess H2O2. The pH optimum of PRXa for the reaction with H2O2 and guaiacol was 5.0 and its specific activity 61 mkat/g protein. Among various aromatic compounds, coniferyl alcohol was polymerized by PRXa to presumed lignin-like material. The extracellular localization and high affinity of PRXa for the cinnamic acid derivatives suggest that this isoenzyme functions in the polymerization or cross-linking of lignin in the plant cell wall.

Planta, 1996

Certain single cells in carrot (Daucus carota L.) suspension cultures react with the monoclonal a... more Certain single cells in carrot (Daucus carota L.) suspension cultures react with the monoclonal antibody JIM8, and it has been proposed that these cells represent a transitional stage in somatic embryo formation. Shortly after isolation of the single cells by sieving, up to 80% of the cells react with JIM8. Within 4 d, JIM8 labelling becomes restricted to 1% of the single cells. To obtain evidence for the proposed correlation between expression of the JIM8 cell wall epitope and somatic embryo formation the developmental fate of carrot single cells labelled with JIM8 was determined by cell tracking. The results, obtained by recording 43 000 cells, show that only few JIM8-1abelled cells give rise to embryos, and most somatic embryos develop from cells devoid of the JIM8 cell wall epitope. We therefore conclude that the presence of the JIM8 cell wall epitope does not coincide with the ability of single suspension cells to form embryos.