Laurel wilt, caused by Raffaelea lauricola, threatens native and non-native species in the Lauraceae in the south-eastern USA. Avocado, Persea americana, is the most important agricultural suscept of laurel wilt. Grafted plants (clonal scions on seedling rootstocks) of 24 cultivars were screened against the disease in the field from 2008 to 2010. Disease was induced with either mycelial plugs or conidial suspensions of R. lauricola. There were significant differences in the severity of disease that developed on different cultivars, and West Indian cultivars were most susceptible (P < 0AE05). Simmonds, a West Indian cultivar that comprises 35% of the commercial production in Florida, was consistently susceptible and was used as a standard genotype in different studies. Disease severity increased significantly on cv. Simmonds as plant size (stem diameter) increased (P < 0AE0042). In greenhouse studies, internal (sapwood) and external disease severities on cv. Simmonds were correlated (P < 0AE0001), and a threshold was evident, in that external symptoms developed only after moderately severe symptoms had developed internally. Latent infection was uncommon; R. lauricola was usually isolated on a semiselective medium or detected via qPCR only from discoloured xylem of inoculated cv. Simmonds. As basipetal movement of the pathogen was common, its movement among trees via root grafts is probable. Greater understanding is needed of the movement of R. lauricola in naturally and artificially infected trees, and whether sufficient tolerance exists in avocado to assist in the management of this important new disease.

The citrus disease huanglongbing (HLB) has become endemic in Florida, with estimates that greater than 80% of citrus trees are currently infected. Although there are no commercial citrus varieties with strong HLB resistance, some field tolerance has been observed in trees exposed to the disease after they were mature. There is great urgency to identify citrus which may permit economic citrus production where HLB is endemic. Therefore, the objective of this study was to assess field tolerance to HLB. To expedite the trial due to urgency, nursery trees were purchased on rootstock varieties as available. The trial included the following unbalanced scion/rootstock combinations: ‘Hamlin/Kinkoji’, ‘Hamlin/Cleopatra’, ‘Temple/Cleopatra’, ‘Fallglo/Kinkoji’, ‘Sugar Belle/Sour Orange’, ‘Tango/Kuharske’, and ‘Ruby Red/Kinkoji’, with most comparisons based solely on scion/rootstock combinations. A randomized complete block experiment was established at Fort Pierce, FL, in Sept. 2010. All trees exhibited symptoms of HLB and tested positive for the Candidatus Liberibacter asiaticus (CLas) bacterium by Oct. 2012, with similar titers [directly assessed as cycle threshold (Ct) using quantitative polymerase chain reaction (qPCR)] measured for all scion/rootstocks at most sample dates, but early titer development in ‘Ruby Red/Kinkoji’ was significantly lower than several other scion/rootstocks. Across all time-points, ‘Fallglo/Kinkoji’ had the lowest rating of distinctive HLB mottling and ‘Ruby Red/Kinkoji’ had the highest rating, but ‘SugarBelle/Sour Orange’ had the highest percentage of leaves affected. After 5 years, ‘SugarBelle/Sour Orange’ and ‘Tango/Kuharske’ had the greatest overall increase in trunk diameter, and were among the healthiest in overall appearance. In Oct. 2015, ‘SugarBelle/Sour Orange’ and ‘Tango/Kuharske’ trees had significantly greater fruit load (80–88 fruit/tree) followed by ‘Temple/Cleopatra’ and ‘Fallglo/Kinkoji’ (31–35 fruit/tree) while ‘Hamlin/Kinkoji’, ‘Hamlin/Cleopatra’, and ‘Ruby Red/Kinkoji’ produced less than 20 fruit per tree. Despite becoming infected by CLas in less than 2 years after planting, the trees continued to grow and all scion/rootstocks displayed increasing fruit production, although very low in ‘Hamlin/Kinkoji’, ‘Hamlin/Cleopatra’, and ‘Ruby Red/Kinkoji’. Growth and fruit production in the highest performing scion/rootstocks were likely less than would be expected for healthy trees, but these results are promising, with markedly better response of some scion/rootstocks with mandarin hybrid scions compared with trees with sweet orange or grapefruit scions. Larger fully replicated trials are underway. It is noteworthy that the most pronounced HLB symptoms and higher early pathogen titer, which are the two criteria most widely used in assessing HLB resistance, were not associated with the lowest growth and cropping, and focus on early symptomatic traits when screening for resistance may obscure important disease tolerance.

Laurel wilt, caused by Raffaelea lauricola, is a lethal, vascular disease of avocado, Persea americana. Its impact on xylem function was examined in artificially inoculated plants of the ÔSimmondsÕ cultivar. Three, 7, 14, 21 and 42 days after inoculation (dai), plants were rated externally and internally for disease severity on a subjective one (asymptomatic) to 10 (dead or 100% symptomatic) scale. Stems were then cut under water, 15 cm below the inoculation point, and placed in 0.1% aqueous solutions of acid fuschin in a greenhouse. After 48 h, the percentage of functional xylem was estimated 10 and 5 cm above and below the inoculation point by quantifying acid fuschin-stained portions of digitized stem cross-sections. Hydraulic conductivity was determined by placing the proximal end of 5-cm-long stem sections, harvested between 5 and 10 cm above the inoculation point, in water and quantifying the volume of water that was drawn over time through the distal end under partial pressure (350 mm Hg). Functional xylem decreased by 3 dai, well before the development of vascular discoloration (7 dai) and wilting of foliage (14 dai). By 14 dai, extensive vascular discoloration had developed and there was a dramatic reduction in functional xylem; plants with internal disease severities of 7 or greater had <20% functional xylem. Hydraulic conductivity decreased exponentially as non-functional xylem and disease severity increased. In plants with internal severities >7, mean flow rates of water were 0.07 ml )1 min )1 cm )2 vs. 42 ml )1 min )1 cm )2 in mock-inoculated plants. The rapid development of these changes suggests that it may be difficult to manage laurel wilt in avocado once plants are infected by R. lauricola. Better understanding of the temporal and spatial development of infection and how the host responds to infection may assist efforts to select laurel wilt-tolerant avocado cultivars.

Raffaelea lauricola causes laurel wilt of avocado, Persea americana. Host × pathogen interactions were examined with light and scanning electron microscopy. The susceptible avocado cultivar 'Simmonds' was inoculated and examined 5 cm above the inoculation site 3, 7, 14, 21, and 42 days after inoculation (dai). No external symptoms were observed at 3 and 7 dai, and there were no anatomical differences when compared with the mock-inoculated plants. By 14 dai, external symptoms were present and dark discoloration had developed in sapwood. Tylose development increased significantly by 14 dai, and was positivity correlated with disease severity (P < 0.05). By 14 dai, gels formed in xylem vessels, fibers, and adjacent parenchyma cells; they were associated with xylem blockage and composed of phenols, pectin, and lipids, as suggested by, respectively, toluidine blue O, ruthenium red, and Sudan III stains. With a chitin-specific stain, fluoresceinconjugated wheat germ agglutinin, infrequent mycelia, and conidia of R. lauricola were visualized within xylem lumena and fibers, regardless of sample date. Understanding how avocado responds to the presence of this pathogen could assist the development of laurel wilt-resistant avocado genotypes and inform efforts to manage this disease with other measures.

Laurel wilt, caused by the fungus Raffaelea lauricola, affects the growth, development, and productivity of avocado, Persea americana. This study evaluated the potential of visible-near infrared spectroscopy for non-destructive sensing of this disease. The symptoms of laurel wilt are visually similar to those caused by freeze damage (leaf necrosis). In this work, we performed classification studies with visible-near infrared spectra of asymptomatic and symptomatic leaves from infected plants, as well as leaves from freeze-damaged and healthy plants, both of which were non-infected. The principal component scores computed from principal component analysis were used as input features in four classifiers: linear discriminant analysis, quadratic discriminant analysis (QDA), Naïve-Bayes classifier, and bagged decision trees (BDT). Among the classifiers, QDA and BDT resulted in classification accuracies of higher than 94% when classifying asymptomatic leaves from infected plants. All of the classifiers were able to discriminate symptomatic-infected leaves from freeze-damaged leaves. However, the false negatives mainly resulted from asymptomatic-infected leaves being classified as healthy. Analyses of average vegetation indices of freeze-damaged, healthy (non-infected), asymptomatic-infected, and symptomatic-infected leaves indicated that the normalized difference vegetation index and the simple ratio index were statistically different.