Use of Coniothyrium minitans as a biocontrol agent and some molecular aspects of sclerotial mycoparasitism

Mitochondria contain their own genome, the integrity of which is required for normal cellular energy metabolism. Reactive oxygen species (ROS) produced by normal mitochondrial respiration can damage cellular macromolecules, including mitochondrial DNA (mtDNA), and have been implicated in degenerative diseases, cancer, and aging. We developed strategies to elevate mitochondrial oxidative stress by exposure to antimycin and H(2)O(2) or utilizing mutants lacking mitochondrial superoxide dismutase (sod2Delta). Experiments were conducted with strains compromised in mitochondrial base excision repair (ntg1Delta) and oxidative damage resistance (pif1Delta) in order to delineate the relationship between these pathways. We observed enhanced ROS production, resulting in a direct increase in oxidative mtDNA damage and mutagenesis. Repair-deficient mutants exposed to oxidative stress conditions exhibited profound genomic instability. Elimination of Ntg1p and Pif1p resulted in a synergistic corruption of respiratory competency upon exposure to antimycin and H(2)O(2). Mitochondrial genomic integrity was substantially compromised in ntg1Delta pif1Delta sod2Delta strains, since these cells exhibit a total loss of mtDNA. A stable respiration-defective strain, possessing a normal complement of mtDNA damage resistance pathways, exhibited a complete loss of mtDNA upon exposure to antimycin and H(2)O(2). This loss was preventable by Sod2p overexpression. These results provide direct evidence that oxidative mtDNA damage can be a major contributor to mitochondrial genomic instability and demonstrate cooperation of Ntg1p and Pif1p to resist the introduction of lesions into the mitochondrial genome.

Biological control agents (BCAs) are potential alternatives for the chemical fungicides presently used in agriculture to fight plant diseases. Coniothyrium minitans is an example of a promising fungal BCA. It is a naturally occurring parasite of the fungus Sclerotinia sclerotiorum, a wide-spread pathogen which substantially reduces the yield of many crops. This review describes, exemplified by C. minitans, the studies that need to be carried out before a fungal BCA is successfully introduced into the market. The main aspects considered are the biology of C. minitans, the development of a product by mass production of spores using solid-state fermentation technology, its biocontrol activity and marketing of the final product.

ABSTRACT The effect of the fungal mycoparasite Coniothyrium minitans applied as a spray to crops infected with Sclerotinia sclerotiorum (causal agent of white mold) on contamination of soil with S. sclerotiorum sclerotia was studied in a 5-year field experiment. Sclerotial survival also was monitored during two subsequent years, when the field was returned to commercial agriculture. In a randomized block design, factorial combinations of four crops and three treatments were repeated 10 times. Potato (Solanum tuberosum), bean (Phaseolus vulgaris), carrot (Daucus carota), and chicory (Cichorium intybus), which are all susceptible to S. sclerotiorum, were grown in rotation. Plots were treated with C. minitans or Trichoderma spp. or were nontreated (control). Crops were rotated in each plot, but treatments were applied to the same plot every year. After 3 years during which it showed no effect on sclerotial survival, the Trichoderma spp. treatment was replaced by a single spray with C. minitans during the fourth and fifth years of the trial. The effect of treatments was monitored in subsequent seasons by counting apothecia as a measure of surviving S. sclerotiorum sclerotia and scoring disease incidence. Trichoderma spp. did not suppress S. sclerotiorum, but C. minitans infected at least 90% of S. sclerotiorum sclerotia on treated crops by the end of the each season. C. minitans lowered the number of apothecia compared with the other treatments during the second year after the bean crop. C. minitans reduced the number of apothecia by approximately 90% when compared with the control and Trichoderma spp. treatments and reduced disease incidence in the bean crop by 50% during the fifth year of the trial, resulting in a slightly higher yield. In 1993, but not 1994, a single spray with C. minitans was nearly as effective at reducing apothecia as three sprays (monitored in 1995). The final population size of sclerotia in soil at the end of the 7-year period was lower in all C. minitans plots than at the beginning of the trial, even in plots where two highly susceptible bean crops were grown during the period. The results indicate that the mycoparasite C. minitans has the potential to keep contamination of soil with sclerotia low in crop rotations with a high number of crops susceptible to S. sclerotiorum.