The first hypothesis was rejected because in many young orchards with severe and uniform occurrences of band canker, detection of any obvious source/s of inoculum within the proximity of these orchards was not present.
Therefore, we focused our research on the second hypothesis: infections may pre-exist in the young trees as latent infections, and after planting the trees in the field these infections develop to disease with apparent symptoms (gummosis, cankers, and tree death).
Previously it was very difficult to identify or isolate the pathogens using a conventional plant pathological methodology. However, we designed and developed a very sensitive and efficient analytical method (qPCR assay) by using pathogen-specific DNA primers and a specific protocol for DNA extraction from plant tissues to target the specific pathogen groups.1 Using this analytical tool, we were able to study and figure out where these initial infections of the Botryosphaeriaceae pathogens originated and how the disease can develop in such young orchards.
To make this determination, we collected healthy looking (symptomless) almond shoots from first, second, and third-leaf orchards and, using the qPCR assay, we detected latent infections even in newly emerged and one-year-old almond shoots (Figure 5). We also found the existence of some of these pathogens in newly emerged and one-year-old shoots, which supports the hypothesis that symptomless trees have latent canker pathogen/s infections by the time they are planted in the commercial field.
Figure 5: Incidence of latent infections of young symptomless almond shoots of different age orchards, using molecular technique (quantitative polymerase chain reaction, qPCR) that quantifies the DNA of specific canker pathogens.
When trees from two nurseries were processed, three fungi (two causing band canker) and a species of Cytospora (a pathogen causing Cytospora canker) were detected in very high levels (Figure 6). The nursery industry takes all kinds of proactive measures, including treating nursery stock material with fungicide, to reduce any type of infection.
These results suggest that additional treatments may be required, first to improve the methodology and treatments done in the nurseries and second to introduce new cultural practices that suppress disease development and protect the newly planted trees in the orchard.
Figure 6: Latent infection levels can vary among different cultivars and nurseries, and in some cases can be very high (i.e., nursery 1).
Similar levels of these fungi were also detected in budwood collected directly from mother almond trees. When processed with the qPCR assay, budwood obtained from two nurseries showed DNA detection (and thus the latent infections) of Cytospora spp., Neofusicoccum spp., and Phomopsis spp. at different levels among nurseries and varieties.
These results imply the possible risk of budwood material that may be carrying these pathogens and can cause canker diseases in young trees after being planted in the field. However, these findings should be further confirmed and compared with large numbers of conventional isolations, using multi-year assessments and working with more nurseries.
Disease management strategy
Based on our findings and the mechanism of band canker epidemics in young almond orchards, our team designed disease management strategies for early in-season applications that take into account the age of trees.
To test our hypothesis, we sprayed an effective fungicide on tissues where latent infections were located to see whether this fungicide could prevent the latent infections from developing disease symptoms. Previous studies showed that Topsin M 70WP (a.i. thiophanate methyl) fungicide was very effective against canker fungi. Therefore, young almond trees in a second leaf orchard were sprayed in early March at a 1.5 lbs. per acre. The spray was done mainly to sufficiently cover the trunk and main scaffolds of these young trees.
The disease was then evaluated in November (about six months after the spray) and showed that the application of Topsin M 70WP significantly reduced the incidence of band canker disease when compared to the untreated trees (Figure 7).
Figure 7: Left: Effect of single fungicide spray applied early March on young almond trees before the appearance of any disease symptoms [disease recording after 10 months]. Right: Effect of single fungicide spray at 15 months after treatment. Each replication includes 50 trees.
These results indicate that when Topsin-M is sprayed on the trunks of young almond trees – before the expression of any symptoms of band canker disease develop – growers can obtain a significant reduction in the incidence of band canker up to 10 months after the applications.
However, the increased incidence of band canker on treated trees recorded 15 months after Topsin-M applications suggests that one spray may not be enough to control the disease. Instead, annual sprays in early spring may be needed to maintain suppression of the disease at low levels and to protect the trees from external infections.
The Almond Board will continue to report on new research and findings related to band canker as they become available. If you have any immediate questions, please contact Dr. Michailides at tjmichailides@ucanr.edu.
1 Specific pathogen groups include Botryosphaeria dothidea, Diplodia spp., Lasiodiplodia spp., Neofusicoccum spp., and Phomopsis spp. (Luo et al., 2017).