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Biomedical subjects

Jonathan Gressel

Publications and source records attributed to Jonathan Gressel.

5 recordsLinked to original sources

Ultralow calcium requirements of fungi facilitate use of calcium regulating agents to suppress host calcium-dependent defenses, synergizing infection by a mycoherbicide.

Infection by many fungi activates a variety of calcium dependent defenses in the hosts, slowing or suppressing the attacker and limiting the efficacy of mycoherbicides. The calcium requirement for fungal growth is so low that it could only be implied based on fungi containing calcium-dependent signaling enzymes. Analytical grade media contain <2 microM calcium, and the addition of specific chelators does not affect fungal growth. Hydrophobic derivatives of the calcium-specific chelator BAPTA designed to traverse plant cuticles were synthesized in order to chelate calcium internally during fungal attack. Some chelators as well as calcium precipitating oxalate and channel blocker verapamil were applied with a weakly mycoherbicidal Colletotrichum coccodes to cotyledons of compatible Abutilon threophrasti. They suppressed calcium dependent callose biosynthesis in the weed and increased virulence but may have affected other calcium-dependent processes that facilitate virulence. The low calcium requirement of fungi, and their high affinity for calcium, allows the application of calcium-regulating agents as synergists for mycoherbicides where the weed uses calcium-dependent defenses.

Calcium↗

Engineering hypervirulence in a mycoherbicidal fungus for efficient weed control.

Agents proposed for biocontrol of major weeds in arable row-crop agriculture have not met expectations because an evolutionary balance has developed between microorganism and weed, even when the mycoherbicide is used inundatively at very high levels (>10(4)spores/cm<(2)). Sufficient virulence can be achieved by transferring genes to the microorganism, tipping the evolutionary balance. Virulence was increased ninefold and was more rapidly effected; furthermore, the requirement for a long duration at high humidity was decreased by introducing NEP1 encoding a phytotoxic protein, to an Abutilon theophrasti-specific, weakly mycoherbicidal strain of Colletotrichum coccodes. The parent strain was at best infective on juvenile cotyledons of this intransigent weed. The transgenic strain was lethal through the three-leaf stage, a sufficient time window to control this asynchronously germinating weed. Strategies of coupling virulence genes with fail-safe mechanisms to prevent spread (due to broadened host range) and to mitigate transgene introgression into crop pathogens could be very useful in the biocontrol of major weeds in row crops.

Agriculture↗

Transformation of carrots with mutant acetolactate synthase for Orobanche (broomrape) control.

Parasitic Orobanche spp are major constraints to vegetable crop production in the Mediterranean basin (to eastern Europe) and in localized places in India, China and the USA. Transgenic target-site herbicide resistance (eg, to acetolactate synthase inhibitors) allows for movement of unmetabolized herbicide through the crop to the photosynthate sink in the parasite, as well as through the soil. We report the successful engineering of a mutant acetolactate synthase (ALS) gene into carrot, allowing control of broomrape already in heterozygotes of the first back-crossed generation, by imazapyr, an imidazolinone ALS inhibitor. It is expected that homozygotes will have higher levels of resistance.

Acetolactate Synthase↗

Universal inheritable barcodes for identifying organisms.

The needs for recognition of novel conventional or transgenic organisms include protection of patented or Identity Preserved lines, detecting transgenics and tracing dispersal. We propose simple 'Biobarcodes' using universal PCR primers to recognize the universal 'nonsense' recognition site of all biobarcodes, followed by a variable nonsense sequence. The proposed sequences are long enough to allow recognition in spite of mutations, have stop codons to prevent coding, and will not self anneal. Sequences of PCR-amplified biobarcodes can be compared to a universal database.

DNA, Plant↗

Infection of tubercles of the parasitic weed Orobanche aegyptiaca by mycoherbicidal Fusarium species.

Progression of the infection by host-specific strains of Fusarium oxysporum and Fusarium arthrosporioides of Orobanche aegyptiaca (Egyptian broomrape) tubercles attached to tomato roots was tracked using light, confocal and electron microscopy. Mycelia transformed with the gene for green fluorescent protein were viewed using a confocal microscope. Fungal penetration was preceded by a rapid loss of starch, with approx. 10 % remaining at 9 h and no measurable starch at 24 h. Penetration into the Orobanche tubercles began by 12 h after inoculation. Hyphae penetrated the outer six cell layers by 24 h, reaching the centre of the tubercles by 48 h and infecting nearly all cells by 72 h. Most of the infected tubercles were dead by 96 h. Breakdown of cell walls and the disintegration of cytoplasm in and around the infected cells occurred between 48 and 96 h. Lignin-like material increased in tubercle cells of infected tissues over time, but did not appear to be effective in limiting fungal penetration or spread. Callose, suberin, constitutive toxins and phytoalexins were not detected in infected tubercles, suggesting that there are no obvious defence mechanisms to overcome. Both Fusarium spp. pathogenic on Orobanche produced fumonisin-like ceramide synthase inhibitors, while fusaric acid was produced only by F. oxysporum in liquid culture. The organisms do not have sufficient virulence for field use (based on glasshouse testing), suggesting that virulence should be transgenically enhanced or additional isolates sought.

Fusarium↗