Mushroom Virus X affected
crop.Mushroom Virus X affected crop.
characterise the interactions between a three A. bisporus strains with different vegetative compatibilities (as this would be an important trait for a virus-breaker strain) as well as characterising the response of a number of strains to virus infection. At the same time, we also wanted to visualise where viruses were located within the mycelium. Having multiple genome sequences for multiple A. bisporus strains is key if you wish to understand the genetic basis that confers an advantage to a particular strain, therefore we also fully sequenced a wild A. bisporus strain to provide new genome data for future research. The experiments and key results are summarised below and will add to the A. bisporus knowledge base for this important crop species, identifying useful traits for future strain development endeavours.
Interaction studies
Three strains of A. bisporus were used in this study: (1) commercial strain A15, (2) a novel experimental fourth-generation hybrid strain (CWH) and (3) a wild strain ARP23 from the ARP collection, all obtained from Sylvan Inc., France. Co-cultures of different strains were set up on agar plates and grown for two weeks at 25°C to allow enough time for ample hyphal interactions to occur (Figure 1). Proteins were
extracted from mycelial tissue and identified using Liquid Chromatography–Mass Spectro- metry (LC-MS/MS). Comparative proteomic analysis was carried out with particular focus on proteins which were significantly more or less abundant between pairwise comparisons. With respect to mycelial interactions between different strains, as expected there was a com- patible interaction between A15-A15, and this manifested as high levels of carbohydrate metabolism in the form of cell wall growth in our comparative proteomics results. The A15- CWH interaction appeared less antagonistic and more representative of a competitive response for substrate, suggesting that A15 and CWH are not interacting very much. The A15-ARP23 interaction on the other hand displayed a high level of vegetative incompatibility and antago- nism, represented by high levels of stress proteins in A15 and abnormal fluffy growth (Figure 1) (O’Connor et al. 2020a). Despite the very different compatibilities bet- ween these three A. bisporus strains, none- theless anastomosis (hyphal fusion) did occur in-vitro between A15 and the other strains. Even low levels of anastomosis are likely to facilitate the transmission of viruses between apparently ‘non-compatible’ strains, increasing the challenge to develop a virus breaker strain.
Ò MUSHROOM BUSINESS 33
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