Biomedical subjects

J Gressel

Publications and source records attributed to J Gressel.

At least 19 recordsLinked to original sources

PubMed2003

Low frequency transmission of a plastid-encoded trait in Setaria italica.

It has been claimed that engineering traits into the chloroplast will prevent transgene transmission by pollen, precluding transgene flow from crops. A Setaria italica (foxtail or birdseed millet) with chloroplast-inherited atrazine resistance (bearing a nuclear dominant red-leaf base marker) was crossed with five male-sterile yellow- or green-leafed herbicide susceptible lines. Chloroplast-inherited resistance was consistently pollen transmitted at a 3x10(-4 )frequency in >780,000 hybrid offspring. The nuclear marker segregated in the F(2), but resistance did not segregate, as expected. Pollen transmission of plastome traits can only be detected using both large samples and selectable genetic markers. The risk of pollen transmission at this frequency would be several orders of magnitude greater than spontaneous nuclear-genome mutation-rates. Chloroplast transformation may be an unacceptable means of preventing transgene outflow, unless stacked with additional mechanisms such as mitigating genes and/or male sterility.

Cell Nucleus↗

PubMed2001

Potential failsafe mechanisms against the spread and introgression of transgenic hypervirulent biocontrol fungi.

Microbial biocontrol agents are typically inefficient owing to the evolutionary necessity to be in balance with their hosts to survive. If transgenetically rendered hypervirulent, however, they could be competitive alternatives to pesticides. Potential means are delineated to prevent, contain or mitigate uncontrollable spread of hypervirulent biocontrol organisms, mutations that increase their host range, and the sexual or asexual introgression of hypervirulence genes into pathogens of other organisms. The use of asporogenic deletion mutants as a platform for generating transgenic hypervirulent biopesticides would prevent such spread. Hypervirulence genes flanked with available 'transgenetic mitigator' (TM) genes (genes that are neutral or positive to the biocontrol agent but deleterious to recombinants) would decrease virulence to non-target species.

Fungi↗

PubMed2000

Transient, oxidant-induced antioxidant transcript and enzyme levels correlate with greater oxidant-resistance in paraquat-resistant Conyza bonariensis.

The elucidation of mechanisms plants use to overcome oxidative stress is facilitated where there is intra-specific genetic variability. The differential induction of higher levels of mRNAs, cytosol and chloroplast antioxidant enzyme activities, and proteins occurred after sub-lethal paraquat treatment of the oxidant-resistant biotype of Conyza bonariensis (L.) Cronq. By 6 h after sub-lethal paraquat treatment the activities of superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11), dehydroascorbate reductase (EC 1.8.5), monodehydroascorbate reductase (EC 1.6.5.4), and glutathione peroxidase (EC 1.11.19) had increased, peaking at 24 h and then slowly reverting back to the basal level. Similarly, the levels of mRNAs encoding these enzymes were enhanced by 12 h and peaked at 18-24 h after sub-lethal paraquat treatment. The time courses of the transient elevation of both transcript and antioxidant enzyme levels correlated with a further transient 2.5- to 3.0-fold increase of paraquat resistance, which occurred only in the constitutively resistant biotype. The individual enzymes seem to be part of a coordinately controlled oxidant tolerance in the resistant biotype, utilizing oxidant-induced, increasingly abundant transcript levels, upon which more antioxidant enzymes were synthesized.

Ascorbate Peroxidases↗

PubMed2000

Molecular biology of weed control.

The vast commercial effort to utilize chemical and molecular tools to solve weed control problems has had a major impact on the basic biological sciences as well as benefits to agriculture, and the first generation of transgenic products has been successful, while somewhat crude. More sophisticated products are envisaged and expected. Biotechnologically-derived herbicide-resistant crops have been a considerable benefit, yet in some cases there is a risk that the same useful transgenes may introgress into related weeds, specifically the weeds that are hardest to control without such transgenic crops. Biotechnology can also be used to mitigate the risks. Molecular tools should be considered for weed control without the use of, or with less chemicals, whether by enhancing crop competitiveness with weeds for light, nutrients and water, or via allelochemicals. Biocontrol agents may become more effective as well as more safe when rendered hypervirulent yet non-spreading by biotechnology. There might be ways to disperse deleterious transposons throughout weed populations, obviating the need to modify the crops.

Crops, Agricultural↗

PubMed1999

Tandem constructs: preventing the rise of superweeds.

Transgenic crops may interbreed with nearby weeds, increasing their competitiveness, and may themselves become a 'volunteer' weed in the following crop. The desired transgene can be coupled in tandem with genes that would render hybrid offspring or volunteer weeds less able to compete with crops, weeds and wild species. Genes that prevent seed shatter or secondary dormancy, or that dwarf the recipient could all be useful for mitigation and may have value to the crop. Many such genes have been isolated in the past few years.

Crops, Agricultural↗

PubMed1997

A high-sensitivity, single-gel, polyacrylamide gel electrophoresis method for the quantitative determination of glutathione reductases.

A new method is described that allows the selective staining and quantification of gluthathione reductases (EC 1.6.4.2) in cell extracts following acrylamide gel electrophoresis. The method is based on modifications of two previous procedures; it uses DTNB [5,5'-dithiobis(2-nitrobenzoic acid)] to develop a yellow color on reaction with GSH formed from the NADPH-dependent reduction of oxidized glutathione. This is followed by specific counterstaining of glutathione reductase with dichlorophenolindophenol/nitroblue tetrazolium. The use of DTNB in the initial staining step inhibits enzymes other than glutathione reductase that could be stained with the dichlorophenolindophenol/nitroblue tetrazolium counterstain. Enzymes such as thioredoxin reductase, which can directly reduce DTNB with NADPH, may be nonselectively stained by this new procedure. Plant ferredoxin-thioredoxin reductase is not reduced by NADPH and therefore does not appear. Glutathione reductase stains much quicker with DTNB in the presence of GSSG than with thioredoxin reductase, allowing them to be distinguished, if parallel gels are run without GSSG, where the two enzymes react at the same rate. The sequential use of two staining procedures results in distinct, sharp permanent bands that can be used to quantify the activity of glutathione reductase while precluding artifacts generated by the previous methods.

2,6-Dichloroindophenol↗

PubMed1992

Isolation, Purification, and Identification of 2-(p-Hydroxyphenoxy)-5, 7-Dihydroxychromone: A Fungal-Induced Phytoalexin from Cassia obtusifolia.

A single phytoalexin was isolated and purified from 12- to 14-day-old leaves of Cassia obtusifolia L. inoculated with Alternaria cassiae Jurair & Khan. The structure was elucidated by (1)H- and (13)C-nuclear magnetic resonance and mass spectrometry as 2-(p-hydroxyphenoxy)-5,7-dihydroxychromone. The compound was shown to be derived in part from phenylalanine. Radial growth of A. cassiae was inhibited 50% by the compound at 0.3 millimolar. This moderate toxicity is compensated for by the relatively high levels (3 millimolar) accumulated. Phenoxychromones have been previously reported only as constitutive secondary metabolites in Artemisia capillaris Thunb., in which their function is unknown.

Journal Article↗

PubMed1992

Glyphosate Suppression of an Elicited Defense Response : Increased Susceptibility of Cassia obtusifolia to a Mycoherbicide.

The major effort in developing pathogenic fungi into potential mycoherbicides is aimed at increasing fungal virulence to weeds without affecting crop selectivity. Specific suppression of biosynthesis of a phytoalexin derived from the shikimate pathway in Cassia obtusifolia L. by a sublethal dose (50 micromolar) of glyphosate increased susceptibility to the mycoherbicide Alternaria cassiae Jurair & Khan. Glyphosate applied with conidia suppressed phytoalexin synthesis beginning at 12 hours, but not an earlier period 8 to 10 hours after inoculation. The phytoalexin synthesis elicited by fungal inoculation was also suppressed by darkness. The magnitudes of virulence of the mycoherbicide in the dark or with glyphosate in the light were both higher than after inoculation in the light with the same concentration of conidia in the absence of glyphosate. Five times less inoculum was needed to cause disease symptoms when applied with glyphosate than without. Glyphosate did not render A. cassiae virulent on soybean (Glycine max), a crop related to the host. These results suggest that a specific inhibition of a weed's elicited defense response can be a safe way to enhance virulence and improve the efficacy of the mycoherbicide.

Journal Article↗

PubMed1991

UDP-rhamnose:flavanone-7-O-glucoside-2''-O-rhamnosyltransferase. Purification and characterization of an enzyme catalyzing the production of bitter compounds in citrus.

The rhamnosyltransferase catalyzing the production of the bitter flavanone-glucosides, naringin and neohesperidin, was purified to homogeneity. The enzyme catalyzes the transfer of rhamnose from UDP-rhamnose to the C-2 hydroxyl group of glucose attached via C-7-O- of naringenin or hesperetin. To our knowledge this is the first complete purification of a rhamnosyl-transferase. The enzyme from young pummelo leaves was purified greater than 2,700-fold to a specific activity of over 600 pmol/min/mg of protein by sequential column chromatographies on Sephacryl S-200, reactive green 19-agarose, and Mono-Q. The enzyme was selectively eluted from the green dye column with only three other proteins by a pulse of the substrate hesperetin-7-O-glucoside followed by UDP. The rhamnosyltransferase is monomeric (approximately 52 kDa) by gel filtration and electrophoresis. The enzyme rhamnosylates only with UDP-rhamnose. Flavonoid-7-O-glucosides are usable acceptors but 5-O-glucosides or aglycones are not. It is inhibited by 10 microM UDP, its end product, but not by naringin or neohesperidin. Several flavonoid-aglycones at 100 microM inhibited the rhamnosyltransferase; UDP-sugars did not. The Km for UDP-rhamnose was similar with prunin (1.3 microM) and hesperetin-7-O-glucoside (1.1 microM) as substrate. The affinity for the natural acceptor prunin (Km = 2.4 microM) was much higher than for hesperetin-7-O-glucoside (Km = 41.5 microM). The isolation of the gene may enable its use in genetic engineering directed to modifying grapefruit bitterness.

Chromatography↗

PubMed1990

Induction of Trichoderma sporulation by nanosecond laser pulses: evidence against cryptochrome cycling.

An important question in the study of photoreceptor action in morphogenesis is whether the chromophore is unidirectionally photobleached, or whether it is recycled, allowing each receptor molecule to be counted more than once. The common soil fungus Trichoderma harzianum grows vegetatively in the dark and sporulates in response to a pulse of blue or UV-A light. Colonies were grown at 26 degrees C, transferred to 3 degrees C, illuminated with non-saturating light, and then put back at 26 degrees C to sporulate. The fluence-response curves for photoinduction in the cold and at 26 degrees C were identical, indicating that there are no enzymatic transduction processes during irradiation. Regions of the perimeter of dark-grown colonies were given single pulses (maximum duration, 30 ns) at 355 nm with a neodymium laser. We obtained a complete fluence-response curve for the laser pulses, which agreed with data for irradiations in the second to minute range. Photoinduction at 3 degrees C, and validity of Bunsen-Roscoe reciprocity from nanoseconds to minutes, support the hypothesis that the inductive event is a simple first-order photobleaching reaction.

Cold Temperature↗

PubMed1989

Increased Tolerance to Photoinhibitory Light in Paraquat-Resistant Conyza bonariensis Measured by Photoacoustic Spectroscopy and CO(2)-Fixation.

Tolerance to photoinhibition was compared between a paraquat-resistant and a sensitive biotype of Conyza bonariensis (L.). Cronq. Photoinhibitory damage was measured as a decrease in oxygen evolution or energy storage using photoacoustic spectroscopy, or as a decrease of (14)CO(2)-fixation. Prior to exposure to high fluence rates, both biotypes had similar quantum yields of oxygen evolution and energy storage. After exposure to high intensity light, the resistant biotype continued to evolve oxygen and to store energy with a high quantum yield while both energy storage and oxygen evolution were severely reduced in the sensitive biotype. CO(2)-fixation was less rapidly inhibited in the resistant biotype compared to the sensitive one. The data show that the paraquat resistant biotype with its high constitutive levels of the chloroplast localized enzymes of the oxygen detoxification pathway, is also partially protected from photoinhibition. This supports the theory that an enhanced radical scavenging system can give temporary protection against photooxidative damage from a variety of sources.

Journal Article↗

PubMed1989

Flavanone Glycoside Biosynthesis in Citrus: Chalcone Synthase, UDP-Glucose:Flavanone-7-O-Glucosyl-Transferase and -Rhamnosyl-Transferase Activities in Cell-Free Extracts.

Previous indirect evidence suggested that the biosynthesis of flavonoids in Citrus may not proceed via the usual chalcone synthase reaction and that glycosylation occurs during chalcone formation and not afterward, as has been reported in other species. We detected chalcone-synthase and UDP-glucose:flavanone-7-O-glucosyl-transferase activities in cell-free extracts of Citrus. The glucosylated flavanone was further rhamnosylated when exogenous UDP-glucose and NADPH were added to the extract. Chalcone-synthase activity was detected in cell-free extracts derived from young leaves and fruits. Young fruits (2 millimeter diameter) had the highest chalcone synthase activity. UDP-glucose:flavanone-7-O-glucosyl-transferase activity was measured in cell-free extracts derived from young leaves and fruits of Citrus mitis and Citrus maxima. The highest UDP-glucose:flavanone-7-O-glucosyl-transferase activity was found in young C. maxima leaves. These data indicate that Citrus contains a flavonoid pathway similar to that studied in other species.

Journal Article↗

PubMed1988

Catabolism of single ring aromatic acids by four Aspergillus species.

Four species of the genus Aspergillus, viz A. fumigatus, A. japonicus, A. niger and A. terreus, decarboxylated, demethoxylated and ring-cleaved aromatic compounds but to different extents. Decarboxylation of vanillate occurred before ring-cleavage, which preceded the release of 14CO2 from the methoxyl group. A large proportion of labelled carbon from the ring of ferulate and vanillate was found in particulate or trichloracetic acid precipitable material of homogenized fungal mycelium. The four Aspergillus species contained vanillate-inducible protocatechuate-3,4-dioxygenase and catechol-1,2-dioxygenase activities.

Aspergillus fumigatus↗

PubMed1987

Acifluorfen Enhancement of Cryptochrome-Modulated Sporulation following an Inductive Light Pulse.

Acifluorfen enhancement of blue-light mediated phototropism suggested that this diphenyl-ether herbicide augments the light reaction (TY Leong, WR Briggs 1983 Plant Physiol 70: 875-881). The separation of the possible direct interaction of acifluorfen with light reactions from interactions with dark pathways has been elucidated in this paper with Trichoderma harzianum. Acifluorfen at 30 micromolar, given for 5 hours in the growth medium, stimulated the conidiation of Trichoderma in response to blue light without apparently affecting growth. Enhanced conidiation could be elicited by dipping cultures into medium with acifluorfen both before as well as 0.5 hour after inductive blue light. This postphotoinduction stimulation indicates that acifluorfen does not directly augment the effect of light by interacting with cryptochrome(s) in Trichoderma. Instead, acifluorfen most probably interacted with the dark reactions following photoinduction.

Journal Article↗

PubMed1987

Kinetic Analysis of Resistance to Paraquat in Conyza: Evidence that Paraquat Transiently Inhibits Leaf Chloroplast Reactions in Resistant Plants.

Paraquat resistance has been claimed to be due to a sequestration of the herbicide before it reaches chloroplasts. This is based on the sensitivity of photosystem I in isolated thylakoids to paraquat, and autoradiographic analyses showing label from paraquat near veins 4 hours after treatment of a resistant biotype. Conversely, the enzymes of the superoxide detoxification pathway were found to be at constitutively elevated levels in intact class A chloroplasts of the resistant biotype of Conyza bonariensis (L.) Cronq. Evidence is presented here that physiologically active levels of paraquat rapidly inhibit chloroplast function in both the resistant and sensitive biotype, before the first sequestration was visualized. This inhibition is transient (completed in 2 hours) in the resistant biotype and irreversible in the sensitive type. Intact class A chloroplasts of the resistant biotype with or without paraquat are less susceptible to photoinduced membrane damage than the sensitive biotype without paraquat, as measured by ethane evolution. These data support a hypothesis that the ability to prevent superoxide damage keeps the resistant biotype viable while paraquat or its metabolites are being sequestered.

Journal Article↗

PubMed1985

Modified cryptochrome in vivo absorption in dim photosporulation mutants of Trichoderma.

Mutants of Trichoderma harzianum that are defective in blue-light-induced sporulation were induced by nutritional stresses as an alternative to light. These mutants may be modified in the putative photoreceptor pigment "cryptochrome" or in an early transduction component. dim (dimsighted) mutants induced by a short transient stress were mapped into four complementation groups and were chosen for study of pigment deficiencies by in vivo absorption spectroscopy. Mutants rib(-)10 and lys(-)44 in the dimY complementation group had altered in vivo absorption spectra in the blue region. Difference spectra obtained by subtracting dimY spectra from that of the wild type had difference bands with peaks at 455 and 480 nm. The similarity between the in vivo difference spectra and the action spectrum for sporulation in wild-type Trichoderma suggests that the mutants lack cryptochrome or have a defective cryptochrome. The decrease in photoresponse as well as the modification of the action spectrum near 480 nm in a dimY mutant support these suggestions. Both dimY mutants pleiotropically accumulate a yellow water-soluble pigment absorbing at wavelengths lower than the blue maxima of cryptochrome; this yellow pigment may be related to cryptochrome.

Journal Article↗

PubMed1984

Evolution of herbicide-resistant weeds.

The first generation of species-selective phenoxy herbicides went into use at the same time as modern antibiotics, chlorinated hydrocarbon insecticides and new fungicides and rodenticides. No resistance has appeared to the still widely used phenoxy herbicides. However, resistance has developed, in many isolated areas around the world, to s-triazine herbicides and, in a few small areas, to bipyridillium herbicides. The parameters of population genetics that govern herbicide resistance and those that govern resistance of microorganisms and fungi to other xenobiotics are the same: generations, selection pressure and fitness. Even though generation times are longer with plants, a sufficient number has passed for resistance to be apparent. The only special factor that controls the development of herbicide resistance in weeds is the spaced germination of seed throughout many seasons. The reason that resistance has not developed to the phenoxy herbicides is probably because of a low effective selection pressure; germination of susceptible seeds occurs late in the season after the herbicide is biodegraded. The highly persistent triazines, and the monthly used, highly ephemeral bipyridillium, paraquat, have exerted much stronger selection pressures. Different modes of tolerance and resistance seem to have evolved in the same species. Crop and herbicide rotation can considerably delay the possibility of resistance development until it is effectively precluded.

Adaptation, Biological↗

PubMed1984

Electric Currents around Growing Trichoderma Hyphae, before and after Photoinduction of Conidiation.

Electric currents were measured around Trichoderma harzianum (Rifai) hyphae using an extracellular vibrating electrode. A steady current enters growing hyphal tips and along the side of the apical millimeter. In addition, outward currents were detected at about one-ninth of the locations tested, 60 to 150 minutes after illumination but not in dark controls. This sporadic, localized outward current pattern might be an early biophysical response to blue light.

Journal Article↗