Efficacy of Potential Control Agents Against Rosellinia necatrix and Their Physiological Impact on Avocado

Rosellinia necatrix is the causal agent of white root rot (WRR), a fatal disease affecting many woody plants, including avocado (Persea americana). As with other root diseases, an integrated approach is required to control WRR. No fully effective control methods are available, and no chemical or biological agents against R. necatrix have been registered for use on avocado in South Africa. Fluazinam has shown promising results in the greenhouse and field in other countries, including Spain. The current study aimed to investigate the potential of a fumigant, chloropicrin, and biological control agents (B-Rus, Beta-Bak, Mity-Gro, and Trichoderma) against R. necatrix both in vitro and in vivo as compared with fluazinam. In a greenhouse trial, results showed that Trichoderma and B-Rus were as effective as fluazinam at inhibiting R. necatrix in vitro and suppressed WRR symptoms when applied before inoculation with R. necatrix. In contrast, Mity-Gro and Beta-Bak failed to inhibit the pathogen in vitro and in the greenhouse trial, despite application of the products to plants before R. necatrix infection. Fluazinam suppressed WRR symptoms in plants when applied at the early stages of infection, whereas chloropicrin rendered the pathogen nonviable when used as a preplant treatment. Plants treated with Trichoderma, B-Rus, and fluazinam sustained dry mass production and net CO2 assimilation by maintaining the green leaf tissues despite being infected with the pathogen. This study has important implications for the integrated management of WRR.

The Rhizobacterium Pseudomonas alcaligenes AVO110 Induces the Expression of Biofilm-Related Genes in Response to Rosellinia necatrix Exudates

The rhizobacterium Pseudomonas alcaligenes AVO110 exhibits antagonism toward the phytopathogenic fungus Rosellinia necatrix. This strain efficiently colonizes R. necatrix hyphae and is able to feed on their exudates. Here, we report the complete genome sequence of P. alcaligenes AVO110. The phylogeny of all available P. alcaligenes genomes separates environmental isolates, including AVO110, from those obtained from infected human blood and oyster tissues, which cluster together with Pseudomonas otitidis. Core and pan-genome analyses showed that P. alcaligenes strains encode highly heterogenic gene pools, with the AVO110 genome encoding the largest and most exclusive variable region (~1.6 Mb, 1795 genes). The AVO110 singletons include a wide repertoire of genes related to biofilm formation, several of which are transcriptionally modulated by R. necatrix exudates. One of these genes (cmpA) encodes a GGDEF/EAL domain protein specific to Pseudomonas spp. strains isolated primarily from the rhizosphere of diverse plants, but also from soil and water samples. We also show that CmpA has a role in biofilm formation and that the integrity of its EAL domain is involved in this function. This study contributes to a better understanding of the niche-specific adaptations and lifestyles of P. alcaligenes, including the mycophagous behavior of strain AVO110.

Loop-Mediated Isothermal Amplification (LAMP)-Based Turn on Fluorescent Paper (ToFP) Device for Detecting Rosellinia necatrix

White root rot (WRR) disease caused by Rosellinia necatrix, a fungal pathogen, results in severe damage to various fruit trees, decreasing their marketability. Regular monitoring is a major process because the pathogen can remain in the soil around the host for a long time. Loop-mediated isothermal amplification (LAMP) is a highly sensitive and efficient amplification technology of nucleic acids (DNA or RNA) that can be performed at constant temperatures. Thus, it has been spotlighted as a useful tool for detecting several infectious agents. In the present study, LAMP-based Turn-on Fluorescent Paper (ToFP) devices were designed and applied to detect R. necatrix. LAMP conditions were optimized and found to be optimal at a reaction temperature (62 °C) and a reaction time (30 minutes). These reaction conditions were confirmed by applying them to infectious soil samples collected from the field. The limitation of detection was identified as 10 fg of genomic DNA under optimized LAMP conditions. These LAMP-based ToFP devices were generated with easily available stationery materials and the utility of these devices to analyze the LAMP results were confirmed through several experiments on a total of 14 field samples. The results showed that the developed LAMP-based detection system was very sensitive and had the advantages of rapid detection and high availability in the field.