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Biomedical subjects

K Apel

Publications and source records attributed to K Apel.

At least 19 recordsLinked to original sources
PubMed1998

Wounding and chemicals induce expression of the Arabidopsis thaliana gene Thi2.1, encoding a fungal defense thionin, via the octadecanoid pathway.

In seedlings of Arabidopsis thaliana the thionin gene Thi2.1 is inducible by methyl jasmonate, wounding, silver nitrate, coronatine, and sorbitol. We have used a biochemical and genetic approach to test the signal transduction of these different inducers. Both exogenously applied jasmonates and jasmonates produced endogenously upon stress induction, lead to GUS expression in a Thi2.1 promoter-uidA transgenic line. No GUS expression was observed in a coil mutant background which lacks jasmonate perception whereas methyl jasmonate and coronatine but not the other inducers were able to overcome the block in jasmonic acid production in a fad3-2 fad7-2 fad8 mutant background. Our results show conclusively that all these inducers regulate Thi2-1 gene expression via the octadecanoid pathway.

Antifungal Agents↗
PubMed1998

High-level expression of a viscotoxin in Arabidopsis thaliana gives enhanced resistance against Plasmodiophora brassicae.

Viscotoxins are a group of toxic thionins found in several mistletoe species. The constitutive CaMV-omega promoter was used to drive the expression of the viscotoxin A3 cDNA from Viscum album in transgenic Arabidopsis thaliana C24. Lines with high viscotoxin A3 levels in all parts of the plant were selected and tested for resistance against the clubroot pathogen Plasmodiophora brassicae. The transgenic lines were more resistant to infection by this pathogen than the parental line.

Animals↗
PubMed1998

Systemic and local induction of an Arabidopsis thionin gene by wounding and pathogens.

The Arabidopsis Thi2.1 thionin gene was cloned and sequenced. The promoter was fused to the uidA gene and stably transformed into Arabidopsis to study its regulation. GUS expression levels correlated with the steady-state levels of Thi2.1 mRNA, thus demonstrating that the promoter is sufficient for the regulation of the Thi2.1 gene. The sensitivity of the Thi2.1 gene to methyl jasmonate was found to be developmentally determined. Systemic and local expression could be induced by wounding and inoculation with Fusarium oxysporum f sp. matthiolae. A deletion analysis of the promoter identified a fragment of 325 bp upstream of the start codon, which appears to contain all the elements necessary for the regulation of the Thi2.1 gene. These results support the view that thionins are defence proteins, and indicate the possibility that resistance of Arabidopsis plants to necrotrophic fungal pathogens is mediated through the octadecanoid pathway.

Agrobacterium tumefaciens↗
PubMed1998

Differential induction of the Arabidopsis thaliana Thi2.1 gene by Fusarium oxysporum f. sp. matthiolae.

The Arabidopsis thaliana Thi2.1 gene is inducible by necrotrophic fungi through a signal transduction pathway different from that for pathogenesis-related (PR) proteins. We have identified three ecotypes that are susceptible (Col-2, Ler, and Ws) and two ecotypes that are resistant (Mt-0 and Uk-4) to spray inoculation with Fusarium oxysporum f. sp. matthiolae. The Thi2.1 transcript level after infection correlates with resistance, being 5 to 10 times higher in the resistant than in the susceptible ecotypes. The beta-glucuronidase (GUS) expression of a Thi2.1-promoter-uidA fusion (with a promoter derived from Col-2) is on the average almost 10 times higher in the Uk-4 background than in the Col-2 background. This confirms the results obtained by Northern (RNA) blots and indicates that Uk-4, and probably other resistant ecotypes too, might have a more sensitive recognition system for F. oxysporum f. sp. matthiolae or might have a signal transduction system that gives a higher amplification of the original recognition signal. Our results suggest a role of the Thi2.1 gene in resistance against F. oxysporum f. sp. matthiolae and perhaps other necrotrophic fungi.

Arabidopsis↗
PubMed1998

Etioplast differentiation in arabidopsis: both PORA and PORB restore the prolamellar body and photoactive protochlorophyllide-F655 to the cop1 photomorphogenic mutant.

The etioplast plastid type of dark-grown angiosperms is defined by the accumulation of the chlorophyll (Chl) precursor protochlorophyllide (Pchlide) and the presence of the paracrystalline prolamellar body (PLB) membrane. Both features correlate with the presence of NADPH:Pchlide oxidoreductase (POR), a light-dependent enzyme that reduces photoactive Pchlide-F655 to chlorophyllide and plays a key role in chloroplast differentiation during greening. Two differentially expressed and regulated POR enzymes, PORA and PORB, have recently been discovered in angiosperms. To investigate the hypothesis that etioplast differentiation requires PORA, we have constitutively overexpressed PORA and PORB in the Arabidopsis wild type and in the constitutive photomorphogenic cop1-18 (previously det340) mutant, which is deficient in the PLB and Pchlide-F655. In both genetic backgrounds, POR overexpression increased PLB size, the ratio of Pchlide-F655 to nonphotoactive Pchl[ide]-F632, and the amount of Pchlide-F655. Dramatically, restoration of either PORA or PORB to the cop1 mutant led to the formation of etioplasts containing an extensive PLB and large amounts of photoactive Pchlide-F655.

Alleles↗
PubMed1997

AtGRP7, a nuclear RNA-binding protein as a component of a circadian-regulated negative feedback loop in Arabidopsis thaliana.

The endogenous clock that drives circadian rhythms is thought to communicate temporal information within the cell via cycling downstream transcripts. A transcript encoding a glycine-rich RNA-binding protein, Atgrp7, in Arabidopsis thaliana undergoes circadian oscillations with peak levels in the evening. The AtGRP7 protein also cycles with a time delay so that Atgrp7 transcript levels decline when the AtGRP7 protein accumulates to high levels. After AtGRP7 protein concentration has fallen to trough levels, Atgrp7 transcript starts to reaccumulate. Overexpression of AtGRP7 in transgenic Arabidopsis plants severely depresses cycling of the endogenous Atgrp7 transcript. These data establish both transcript and protein as components of a negative feedback circuit capable of generating a stable oscillation. AtGRP7 overexpression also depresses the oscillation of the circadian-regulated transcript encoding the related RNA-binding protein AtGRP8 but does not affect the oscillation of transcripts such as cab or catalase mRNAs. We propose that the AtGRP7 autoregulatory loop represents a "slave" oscillator in Arabidopsis that receives temporal information from a central "master" oscillator, conserves the rhythmicity by negative feedback, and transduces it to the output pathway by regulating a subset of clock-controlled transcripts.

Arabidopsis↗
PubMed1997

ESTs reveal a multigene family for plant defensins in Arabidopsis thaliana.

Plant defensins, formerly named gamma-thionins, are a group of small, cysteine-rich, basic, and antimicrobial plant proteins. Random sequencing of expressed sequence tags (ESTs) in Arabidopsis thaliana has revealed several different plant defensin genes in this plant species which can be grouped into two subfamilies. We have used one EST of each subfamily to study the expression of the corresponding genes in A. thaliana. Pdf2.3 is constitutively expressed in seedlings, rosettes, flowers, and siliques and is not inducible in seedlings either by methyl jasmonate, salicylate, ethephon, and silver nitrate or by several different phytopathogenic fungi. The expression of a second gene, Pdf1.2, is in untreated plants only detectable in rosettes. In seedlings, it is inducible by methyl jasmonate, silver nitrate, and different phytopathogenic fungi, notably Fusarium oxysporum f. sp. matthiolae. The regulation of Pdf1.2 resembles that of the pathogen-inducible thionin gene Thi2.1.

Alternaria↗
PubMed1997

Overexpression of light-dependent PORA or PORB in plants depleted of endogenous POR by far-red light enhances seedling survival in white light and protects against photooxidative damage.

The structurally related light-dependent protochlorophyllide (Pchlide) oxidoreductases PORA and PORB mediate the only light-requiring step in chlorophyll (Chl) biosynthesis in higher plants. Correlative evidence suggests that some in vivo functions of PORA and PORB may be unique, including a postulated photoprotective role for PORA. For example, wild-type Arabidopsis thaliana seedlings grown in non-photooxidative far-red light (cFR) resemble those grown in white light (WL), but they are yellow and do not green normally thereafter in WL. This defect is accompanied by the absence of detectable PORA and reduced levels of PORB expression. Here, direct evidence is provided that the presence of POR, either as PORA or PORB, can confer photoprotection in plants. In contrast to the wild-type, the plastids of transgenic PORA- or PORB-overexpressing Arabidopsis seedlings grown in cFR possess extensive prolamellar bodies. Upon a subsequent shift to WL, POR-overexpressing seedlings develop thylakoid membranes, accumulate large amounts of Chl and are viable at fluence rates lethal to the wild-type. Intriguingly, the plastid membrane architectures of greening transgenic seedlings seem to depend on whether PORA or PORB has been overproduced. POR-overexpressing seedlings shifted from cFR to WL of fluence rates from 20 to 500 muE m-2 sec-1 accumulate substantially higher amounts of Chl than does the wild-type. Furthermore, the WL fluence rate that permits maximal Chl accumulation increases from 8 muE m-2 sec-1 in the wild-type to 125 muE m-2 sec-1 in transgenic seedlings. POR overexpression during growth in cFR also correlates with a fourfold decrease in the steady-state content of Pchlide, a potentially lethal photosensitizer.

Agrobacterium tumefaciens↗
PubMed1997

Overexpression of an endogenous thionin enhances resistance of Arabidopsis against Fusarium oxysporum.

Thionins are antimicrobial proteins that are thought to be involved in plant defense. Concordant with this view, we have recently shown that the Arabidopsis thionin Thi2.1 gene is inducible by phytopathogenic fungi. Here, we demonstrate that constitutive overexpression of this thionin enhances the resistance of the susceptible ecotype Columbia (Col-2) against attack by Fusarium oxysporum f sp matthiolae. Transgenic lines had a reduced loss of chlorophyll after inoculation and supported significantly less fungal growth on the cotyledons, as evaluated by trypan blue staining. Moreover, fungi on cotyledons of transgenic lines had more hyphae with growth anomalies, including hyperbranching, than on cotyledons of the parental line. No transcripts for pathogenesis-related PR-1, PR-5, or the pathogen-inducible plant defensin Pdf1.2 could be detected in uninoculated transgenic seedlings, indicating that all of the observed effects of the overexpressing lines are most likely the result of the toxicity of the THI2.1 thionin. Our findings strongly support the view that thionins are defense proteins.

Antimicrobial Cationic Peptides↗
PubMed1997

FPF1 promotes flowering in Arabidopsis.

We have characterized the gene flowering promoting factor1 (FPF1), which is expressed in apical meristems immediately after the photoperiodic induction of flowering in the long-day plants mustard and Arabidopsis. In early transition stages, expression is only detectable in the peripheral zone of apical meristems; however, later on, it can also be found in floral meristems and in axillary meristems that form secondary inflorescences. The FPF1 gene encodes a 12.6-kD protein that has no homology to any previously identified protein of known function. Constitutive expression of the gene in Arabidopsis under control of the cauliflower mosaic virus 35S promoter resulted in a dominant heritable trait of early flowering under both short- and long-day conditions. Treatments with gibberellin (GA) and paclobutrazol, a GA biosynthesis inhibitor, as well as crosses with GA-deficient mutants indicate that FPF1 is involved in a GA-dependent signaling pathway and modulates a GA response in apical meristems during the transition to flowering.

Amino Acid Sequence↗
PubMed1996

Transcripts of the two NADPH protochlorophyllide oxidereductase genes PorA and PorB are differentially degraded in etiolated barley seedlings.

The light-dependent reduction of protochlorophyllide to chlorophyllide in higher plants is catalyzed by two closely related enzymes, the NADPH-Pchlide oxidoreductases A and B that are encoded by the nuclear genes PorA and PorB, respectively. The expression of the PorA gene is negatively regulated by light. It has formerly been reasoned that, apart from the well-studied transcriptional down-regulation, a post-transcriptional mechanism may exist that contributes markedly to the light-induced decline of PorA mRNA steady-state levels. We investigated the degradation kinetics of the PorA messenger after inhibiting RNA synthesis with cordycepin. The PorA mRNA was found to be inherently unstable. In contrast, the PorB mRNA was shown to be stabilized in the presence of cordycepin, suggesting degradation by a mechanism different from that of PorA mRNA degradation. The PorA messenger instability is postulated to be conferred by a previously described plant-specific DST element in its 3'UTR.

Base Sequence↗
PubMed1996

Identification of two MADS box genes that are expressed in the apical meristem of the long-day plant Sinapis alba in transition to flowering.

To understand the molecular mechanisms involved in the transition from vegetative to reproductive growth, the photoperiodic-controlled induction of flowering in the long-day plant Sinapis alba was used to screen for regulatory genes which are expressed upon induction in the apical meristem. By using the conserved MADS box-encoding region of the organ identity gene AGAMOUS, the genes SaMADS A and SaMADS B were identified which are expressed in transition stages of mustard. RNA blot analysis has confirmed that the transcript levels of both genes are drastically increased shortly after the induction of flowering and that both genes are expressed earlier than the known MADS box genes. In situ hybridization studies have shown that the expression of the genes is restricted to the apical meristem of induced plants during early phases of reproductive development. The expression of SaMADS A is first detectable in the central zone of the meristem, a region where the earliest changes of an evoked meristem could be detected by classical physiological methods, suggesting that SaMADS A may have an important function during the transition to flowering.

Amino Acid Sequence↗
PubMed1996

Distinct roles for light-dependent NADPH:protochlorophyllide oxidoreductases (POR) A and B during greening in higher plants.

Light-dependent NADPH:protochlorophyllide oxidoreductase (POR), a nuclear-encoded plastid-localized enzyme, catalyzes the photoreduction of protochlorophyllide (Pchlide) to chlorophyllide in higher plants, algae and cyanobacteria. Angiosperms require light for chlorophyll (Chl) biosynthesis and have recently been shown to contain two POR-encoding genes, PorA and PorB, that are differentially regulated by light and developmental state. PorA expression rapidly becomes undetectable after illumination of etiolated seedlings, whereas PorB expression persists throughout greening and in adult plants. In order to study the in vivo functions of Arabidopsis POR A and POR B we have abolished the expression of PorA through the use of the phytochrome A-mediated far-red high irradiance response. Wild-type seedlings grown in continuous far-red light (cFR) display the morphology of white light (WL)-grown seedlings, but contain only traces of Chl and do not green upon transfer to WL. This cFR-induced greening defect correlates with the absence of PorA mRNA, the putative POR A protein, phototransformable Pchlide-F655, and with strongly reduced POR enzymatic activity in plant extracts. In contrast, a cFR-grown phyA mutant expresses the PorA gene, accumulates Chl and visibly greens in WL. Furthermore, constitutive overexpression of POR A in cFR-grown transgenic Arabidopsis wild-type seedlings restores Chl accumulation and WL-induced greening. These data demonstrate that POR A is required for greening and that the availability of POR A limits Chl accumulation during growth in cFR. POR B apparently provides a means to sustain light-dependent Chl biosynthesis in fully greened, mature plants in the absence of phototransformable Pchlide-F655.

Arabidopsis↗
PubMed1995

Two routes of chlorophyllide synthesis that are differentially regulated by light in barley (Hordeum vulgare L.).

NADPH-protochlorophyllide oxidoreductase (POR; EC 1.6.99.1) catalyzes the only known light-dependent step in chlorophyll synthesis of higher plants, the reduction of protochlorophyllide (Pchlide) to chlorophyllide. In barley, two distinct immunoreactive POR proteins were identified. In contrast to the light-sensitive POR enzyme studied thus far (POR-A), levels of the second POR protein remained constant in seedlings during the transition from dark growth to the light and in green plants. The existence of a second POR-related protein was verified by isolating and sequencing cDNAs that encode a second POR polypeptide (POR-B) with an amino acid sequence identity of 75% to the POR-A. In the presence of NADPH and Pchlide, the in vitro-synthesized POR-A and POR-B proteins could be reconstituted to ternary enzymatically active complexes that reduced Pchlide to chlorophyllide only after illumination. Even though the in vitro activities of the two enzymes were similar, the expression of their genes during the light-induced transformation of etiolated to green seedlings was distinct. While the POR-A mRNA rapidly declined during illumination of dark-grown seedlings and soon disappeared, POR-B mRNA remained at an approximately constant level in dark-grown and green seedlings. Thus these results suggest that chlorophyll synthesis is controlled by two light-dependent POR enzymes, one that is active only transiently in etiolated seedlings at the beginning of illumination and the other that also operates in green plants.

Amino Acid Sequence↗
PubMed1995

Comparison of different constitutive and inducible promoters for the overexpression of transgenes in Arabidopsis thaliana.

We compared the organ specificity and the strength of different constitutive (CaMV-35S, CaMV-35Somega, Arabidopsis ubiquitin UBQ1, and barley leaf thionin BTH6 promoter) and one inducible promoter (soybean heat-shock promoter Gmhsp17.3) in stably transformed Arabidopsis thaliana plants. For this purpose we constructed a set of plant expression vectors equipped with the different promoters. Using the uidA reporter gene we could show that the CaMV-35S promoter has the highest expression level which was enhanced two- to threefold by the addition of a translational enhancer (TMV omega element) without altering the organ specificity of the promoter. The barley leaf thionin promoter was almost inactive in the majority of lines whereas the ubiquitin promoter exhibited an intermediate strength. The heat-shock promoter was inducible up to 18-fold but absolute levels were lower than in the case of the ubiquitin promoter. Conclusive quantitative results for different organs and developmental stages were obtained by the analysis of 24 stably transformed lines per promoter construct.

Antimicrobial Cationic Peptides↗
PubMed1995

Isolation and classification of chlorophyll-deficient xantha mutants of Arabidopsis thaliana.

Mutant lines of Arabidopsis thaliana that are either blocked at various steps of the biosynthetic pathway of chlorophyll (Chl) or that are disturbed in one of the subsequent steps leading to the assembly of an active photosynthetic membrane were isolated by screening for Chl-deficient xantha (xan) mutants. Only mutants that segregated in a 3:1 ratio, that contained the same carotenoid spectrum as etiolated wild-type seedlings and less than 2% of the Chl of wild-type control seedlings, and whose Chl content was not affected by the addition of sucrose to the growth medium were selected for a more detailed analysis. As a final test for the classification of the selected mutants, light-grown xan mutants were vacuum-infiltrated and incubated with the common precursor of tetrapyrroles, delta-aminolevulinic acid (ALA), in the dark. Two major groups of mutants could be distinguished. Some of the mutants were blocked at various steps of the Chl pathway between ALA and protochlorophyllide (Pchlide) and did not accumulate the latter in the dark. The other mutants accumulated Pchlide in the dark regardless of whether exogenous ALA was added. This latter group could be subdivided into mutants with a biochemical lesion in a recently discovered second light-dependent Pchlide reduction step that occurs in green plants and mutants that have blocks in the assembly of Chl protein complexes. In the present work a total of seven different loci could be defined genetically in Arabidopsis that affect the synthesis of Chl and its integration into the growing photosynthetic membrane.

Arabidopsis↗