Laurel Wilt Research Articles

Quantification of Propagules of the Laurel Wilt Fungus and Other Mycangial Fungi from the Redbay Ambrosia Beetle,Xyleborus glabratus

The laurel wilt pathogen, Raffaelea lauricola, is a fungal symbiont of the redbay ambrosia beetle, Xyleborus glabratus, which is native to Asia and was believed to have brought R. lauricola with it to the southeastern United States. Individual X. glabratus beetles from six populations in South Carolina and Georgia were individually macerated in glass tissue grinders and serially diluted to quantify the CFU of fungal symbionts. Six species of Raffaelea were isolated, with up to four species from an individual adult beetle. The Raffaelea spp. were apparently within the protected, paired, mandibular mycangia because they were as numerous in heads as in whole beetles, and surface-sterilized heads or whole bodies yielded as many or more CFU as did nonsterilized heads or whole beetles. R. lauricola was isolated from 40 of the 41 beetles sampled, and it was isolated in the highest numbers, up to 30,000 CFU/beetle. Depending on the population sampled, R. subalba or R. ellipticospora was the next most frequently isolated species. R. arxii, R. fusca, and R. subfusca were only occasionally isolated. The laurel wilt pathogen apparently grows in a yeast phase within the mycangia in competition with other Raffaelea spp.

Homeowner Detection of and Recommendations for Mitigating Redbay Ambrosia Beetle – Laurel Wilt Disease on Redbay and Avocado Trees in the Home Landscape

HS1179, a 4-page illustrated fact sheet by Jonathan H. Crane and Jason A. Smith, provides homeowners with an update on redbay ambrosia beetle-laurel wilt disease in Florida, how the beetle and pathogen are spread, symptoms of infestations, and recommendations for homeowners. Includes references. Published by the UF Department of Horticultural Sciences, May 2010.

First Occurrence of Laurel Wilt Disease Caused by Raffaelea lauricola on Redbay Trees in Mississippi.

Laurel wilt is a lethal, nonnative vascular wilt disease of redbay (Persea borbonia), sassafras (Sassafras albidum), and other trees in the Lauraceae (1,4). It is caused by a fungus (Raffaelea lauricola) and transmitted by the redbay ambrosia beetle (Xyleborus glabratus), a nonnative insect first detected in Georgia in 2002 (1,2). Since introduction of the pathogen and vector (presumably from Asia), laurel wilt has caused extensive mortality to redbays in Georgia, Florida, and South Carolina (1). In June 2009, a landowner in Gautier, MS reported dead redbay trees. Signs and symptoms were identical to those reported for laurel wilt along the Atlantic Coast (wilted, bronze red foliage, and dark gray-to-black vascular discoloration) (1). Infected trees have subsequently been confirmed in and near the Pascagoula River Basin. Size of infected redbays ranged from 5 to 20 cm (diameter at breast height). No heavily decomposed or fallen redbays were noted. Many individual specimens exhibited extensive drying of stem wood and dry, wilted, light brown foliage. This indicates that introduction to the area may have occurred within the last 3 years. X. glabratus adults were collected (30°26'44.45″N, 88°39'41.83″W) in a Lindgren funnel trap baited with phoebe and manuka oil lures. Beetle identification was confirmed by USDA-APHIS, and voucher specimens were submitted to the Smithsonian National Museum of Natural History and the Mississippi Entomological Museum. Symptomatic redbay wood chips from the same location were surface sterilized and plated on cycloheximide-streptomycin malt agar and R. lauricola was isolated. A 1,026-bp portion of 18S rDNA (GenBank No. GQ996063) was amplified by PCR and sequenced using primers NS1 and NS4. BLASTn searches revealed perfect homology to R. lauricola isolate PL 697 (GQ329704). Two isolates of R. lauricola were recovered and prepared into separate spore suspensions (1 × 108 CFU/ml). Each isolate was inoculated into two healthy redbays. The inoculated redbays were placed in a growth chamber with two water-only controls. All inoculated plants, and none of the controls, exhibited wilt symptoms and died within 20 days. R. lauricola was recovered from the discolored sapwood of the inoculated plants, completing Koch's postulates. A model prediction for the natural dispersion of X. glabratus and R. lauricola estimated that these organisms may not reach Mississippi for 10 to 15 years (3). The current detection of laurel wilt in Mississippi is substantially ahead of this estimate. Currently, no records of laurel wilt have been reported from western Georgia, all of Alabama, or the panhandle of Florida. Confirmed locations in Mississippi are in Jackson County, along the Interstate 10 corridor and the Pascagoula River drainage. Due to the relatively large extent of the infestation (~64 km2, including hundreds of infected trees) eradication is not being attempted. Surveys, remote sensing, and phylogeographic analysis are underway to delineate the extent of infestation and discover the mode of introduction. The current outbreak of laurel wilt in Mississippi is likely the result of human transport of infested wood, either from Asia as a separate, new introduction or from previously infested areas in the southeastern United States.

Potential Economic Impact of Laurel Wilt Disease on the Florida Avocado Industry

This article describes and provides preliminary estimates of the potential economic losses that could result from an incursion of the recently discovered exotic laurel wilt disease caused by Raffaelea lauricola, in the main avocado (Persea americana) growing area of Florida. Estimates are provided for the direct losses as well as the indirect or “spillover” losses that could occur across the rest of the regional economy. The Impact Analysis for Planning (IMPLAN) input-output multipliers were used in assessing the regional impacts. The results of the investigation indicate that the direct loss to the industry in terms of lost sales, property damage, and increased management costs could range from $356 million in a do-nothing situation to about $183 million if damage control measure were 50% effective. If increased management costs and decreased property values are ignored, the adverse impact on the regional economy could range from $54 million in a do-nothing situation to $27 million in a case in which the treatments result in only a 50% reduction in avocado production.

First Report of Laurel Wilt Disease Caused by Raffaelea lauricola on Sassafras in Florida and South Carolina

Laurel wilt disease, caused by Raffaelea lauricola (T.C. Harr., Fraedrich & Aghayeva sp. nov.), which is a fungal symbiont of the nonnative redbay ambrosia beetle (Xyleborus glabratus Eichhoff), has caused widespread mortality of native redbay (Persea borbonia (L.) Spreng) in Georgia, South Carolina, and Florida since 2002. The disease has been noted on other species in the Lauraceae including sassafras in Georgia (1), and more recently, on avocado and camphor in Florida (4). Since 2005, wilted shoots, branch dieback, and tree death have been observed in sassafras trees (Sassafras albidum (L.)) in Liberty, McIntosh, Chatham, Effingham, Bulloch, Evans, and Screven counties in Georgia; Bamberg, Beaufort, Charleston, Colleton, Hampton, and Orangeburg counties in South Carolina; and Putnam County in Florida. Symptomatic sassafras trees ranged from 1 to 12 m high and 2.5 to 25 cm in diameter at breast height. In contrast to red bay trees that retain wilted foliage, symptomatic sassafras defoliate rapidly as trees wilt and die. Multiple symptomatic ramets originating from a common root system have been observed. Removal of bark from stem and root sections from wilted trees revealed black-to-brownish staining in the sapwood, characteristic of laurel wilt. Wood chips from symptomatic areas of branches and roots were surface sterilized and plated on cycloheximide-streptomycin malt agar as previously described (1) and R. lauricola was routinely isolated. Small subunit (18S) sequences from rDNA were amplified by PCR and sequenced using primers NS1 and NS4 (3) for isolates from sassafras from Florida and South Carolina. BLASTn searches revealed homology to Raffaelea sp. C2203 (GenBank Accession No. EU123076, 100% similarity) described by Fraedrich et al. (1) from redbay and later named R. lauricola (2). The small subunit rDNA sequences for these isolates have been deposited into GenBank ( http://www.ncbi.nlm.nih.gov/Genbank/index.html ) and assigned Accession Nos. EU980448 (Florida) and GQ329704 (South Carolina). Koch's postulates have been completed with R. lauricola on this host previously (1). Laurel wilt on sassafras often was geographically isolated from other symptomatic hosts in Georgia and South Carolina and appears to occur on this host independently of proximity to redbay. Further studies to determine the epidemiology of laurel wilt on sassafras, potential resistance, and impact on sassafras life history and distribution are needed. Given the clonal nature of sassafras, the disease would appear to have the potential to move through roots of trees once established in a stand.

First Report of Laurel Wilt Disease Caused by Raffaelea lauricola on Camphor in Florida and Georgia.

Laurel wilt is a recently described (1) vascular disease of redbay (Persea borbonia (L.) Spreng) and other plants in the family Lauraceae. The wilt is caused by Raffaelea lauricola, a fungus vectored by the nonnative redbay ambrosia beetle (Xyleborus glabratus Eichhoff) (1,2). Since 2003, laurel wilt has caused widespread mortality of redbay in Georgia, South Carolina, and Florida (1) and has recently been found on avocado in Florida (4). Since June of 2007, wilted shoots and branch dieback have been observed in several camphor trees (Cinnamomum camphora (L.) Sieb.) in residential areas of McIntosh and Glynn counties in Georgia and Baker County in Florida. Symptomatic camphor trees ranged from 4.5 to 12 m high and occurred in areas where redbay mortality due to laurel wilt has been frequently observed during the last 2 to 3 years. In some camphor trees, only the smaller branches (<2 cm in diameter) were wilting or dead, whereas in other trees (e.g., Baker County, Florida), the larger branches and substantial portions of the crown were also symptomatic. Rapid wilt that affects entire trees that is usually observed in redbay, has not been observed in camphor. Some camphor trees in residential areas of Jekyll Island, Georgia (Glynn County), where extensive wilt of redbay has occurred, have exhibited only localized wilt of some shoots or branches and other camphors remain asymptomatic. Removal of bark from wilted branch sections revealed black-to-brownish staining in the sapwood, characteristic of laurel wilt. Although no evidence of ambrosia beetles was observed on these samples, more extensive surveys are needed to determine the role of this vector in laurel wilt of camphor. Wood chips from symptomatic areas of branches were surface sterilized and plated on cycloheximide-streptomycin malt agar as previously described (1,4) and R. lauricola was routinely isolated. Small subunit (18S) sequences from rDNA were amplified by PCR and sequenced using primers NS1 and NS4 (3). BLASTn searches revealed homology to R. lauricola C2203 (GenBank Accession No. EU123076, 100% similarity, e-value of 0.0, and a total score of 1,886). The small subunit rDNA sequence for this isolate has been deposited into GenBank ( http://www.ncbi.nlm.nih.gov/Genbank/index.html ) and has been assigned Accession No. EU 853303. The presence of laurel wilt in camphor provides an opportunity to understand the pathogen distribution and possible resistance mechanisms in this host, which could have implications for efforts to remediate the impacts of the disease in redbay and other species in the Lauraceae in the southeastern United States.

California Laurel Is Susceptible to Laurel Wilt Caused by Raffaelea lauricola.

Extensive mortality of redbay (Persea borbonia (L.) Spreng.) has been observed in the southeastern United States since 2003. The mortality is due to laurel wilt caused by Raffaelea lauricola T. C. Harr., Fraedrich & Aghayeva, a fungal symbiont of the recently introduced redbay ambrosia beetle (RAB), Xyleborus glabratus Eichhoff (1,2). The wilt is known to affect other members of the Lauraceae including sassafras (Sassafras albidum (Nuttall) Nees) and avocado (Persea americana Mill.) (1,3). Two inoculation experiments were conducted to evaluate the susceptibility of California laurel (Umbellularia californica (Hook. & Arn.) Nutt.) to R. lauricola. Seedlings, averaging 73 cm high and 13 mm in diameter, were wounded with a drill bit (2.8 mm) to a depth of one-half the diameter of the stems. In each experiment, 10 seedlings were inoculated with one of two isolates of R. lauricola (five seedlings per isolate) obtained as previously described (1) from wilted redbays on Hilton Head Island, South Carolina and Fort George Island, Florida. In the first experiment, seedlings were inoculated with spore suspensions (0.1 ml) ranging from 1.9 to 2.3 × 106 spores/ml and produced as previously described (1). In the second experiment, seedlings were inoculated with mycelial plugs obtained from the edge of 10-day-old cultures growing on malt extract agar (MEA). Five seedlings in each experiment served as controls and were inoculated with sterile deionized water or plugs of sterile MEA. Inoculation points were wrapped with Parafilm M (Pechiney Plastic Packaging, Menasha, WI). Seedlings were grown in growth chambers (daytime temperature 26°C, nighttime 24°C, and a 15-h photoperiod) for 13 to 15 weeks. At the end of the first experiment, 7 of 10 seedlings inoculated with R. lauricola exhibited wilt that appeared as a dieback of a few to the majority of branches. Nine of the ten seedlings exhibited sapwood discoloration and the fungus was isolated from eight of these seedlings. At the end of the second experiment, 8 of 10 seedlings exhibited wilt that again appeared as a dieback of a few branches to most branches. All seedlings with wilt exhibited sapwood discoloration and the fungus was recovered from these seedlings. Two seedlings inoculated with R. lauricola exhibited no symptoms of disease and the fungus was not recovered. Control seedlings remained healthy in both experiments with no evidence of wilt or sapwood discoloration and R. lauricola was not isolated. These results indicate that California laurel is susceptible to laurel wilt caused by R. lauricola. Furthermore, the disease on California laurel may appear as a branch dieback affecting individual branches one at a time rather than a rapid wilt of the entire crown as is often observed in redbay (1). Currently, the RAB is not known to occur on the West Coast and it is also not known if this beetle is capable of attacking and producing brood on California laurel. Nonetheless, if the RAB and R. lauricola become established on the West Coast, laurel wilt could pose a serious threat to natural ecosystems as well as the avocado industry in California.

Ability of the Redbay Ambrosia Beetle (Coleoptera: Curculionidae: Scolytinae) to Bore into Young Avocado (Lauraceae) Plants and Transmit the Laurel Wilt Pathogen (Raffaelea sp)

Laurel wilt is a vascular disease of plants in the Laurel family (Lauraceae) that has caused ex tensive mortality of redbay (Persea borbonia (L.) Spreng) trees in South Carolina, Georgia, and Florida. The disease is caused by a previously un described species of Raffaelea, a fungal symbiont of the non-native redbay ambrosia beetle, Xyle borus glabratus Eichhoff (Fraedrich et al. 2008). Xyleborus glabratus was initially detected in the US. near Savannah, GA in 2002 (Rabaglia et al. 2006). The beetle carries spores of the laurel wilt fungus in mandibular mycangia and inoculates the xylem of host trees by boring into the wood. In redbay, the fungus moves rapidly through the xy lem, plugging the flow of water and causing trees to die in a matter of weeks or a few months. Af fected trees are characterized by a dark discolor ation in the outer sapwood (Fraedrich et al. 2008). Laurel wilt continues to devastate populations of redbay in the southeastern coastal plain and the distribution of the disease has rapidly expanded (Johnson et al. 2008). In the summer and fall of 2006, 16 small avo cado (Persea americana var. americana Mill.) trees were planted on Ft. George Island, FL, a site with a high incidence of laurel wilt. By 12 Feb 2007, X glabratus and its associated Raffaelea sp. were obtained from two dying trees of the West In dian cultivar Donnie (A.E.M., J.E.P. & J.H.C., un published data). In September 2007, a large avo cado tree in a homeowner's yard in Jacksonville, FL also was discovered to be diseased with laurel wilt (Mayfield et al. 2008). In growth chamber ex periments, some avocado plants wilted after arti ficial inoculation with the laurel wilt pathogen (Fraedrich et al. 2008). Furthermore, avocado wood has been demonstrated to be attractive toX glabratus in field trapping trials (Hanula et al. in press). Concern exists that laurel wilt and X. gla bratus could negatively impact the commercial avocado industry in south Florida and beyond. There are 3 distinct races of avocado, West In dian (WI) (Persea americana var. americana Mill), Guatemalan (G) (P. americana var. guatemalensis Williams) and Mexican (M) [P. americana var drymifloia (Schlect. and Cham.)]. In Florida, West Indian and West Indian-Guatemalan hybrids are commercially grown; in California, mostly Guate malan-Mexican hybrids are grown. Two no-choice experiments were conducted to determine (1) whether Xyleborus glabratus would bore into healthy young avocado plants of various genetic backgrounds, and (2) whether these bee tles could transmit the laurel wilt pathogen (Raf faelea sp.) into those plants. Stem sections (ap prox. 30 cm long and 10 cm in diameter) from wilted redbay trees infested with X. glabratus were collected from Jennings State Forest, Clay County, FL in early May 2007 and placed in rear ing containers at the Florida DACS Division of Plant Industry in Gainesville, FL. Emerging adult females were collected, sealed in Petri plates and transferred to a quarantine green house at the University of Florida, Indian River Research & Education Center in Fort Pierce, FL. Experimental plants were obtained in 3.8-L pots from a commercial nursery near Homestead, FL and ranged from 1.2-4.4 cm in basal diameter and 43.0-94.0 cm in height. Experiment 1 included 4 plants from each of 4 avocado cultivars represent ing different genetic backgrounds: 'Hass' (G-M hybrid), 'Simmonds' (WI), 'Monroe' (G-WI hybrid), and Winter Mexican' (M). These particular culti vars were of interest because 'Hass' is a widely grown commercial cultivar in California, 'Sim monds' and 'Monroe' are popular Florida culti vars, and 'Winter Mexican' is a pure Mexican race cultivar. Within each cultivar, 2 plants were ran

Effect of Propiconazole on Laurel Wilt Disease Development in Redbay Trees and on the Pathogen In Vitro

Laurel wilt is a vascular disease of Lauraceous plants caused by a fungus (Raffaelea spp.) that is vectored by a recently introduced, nonnative ambrosia beetle (Xyleborus glabratus). The disease is devastating to redbay (Persea borbonia) trees in forests, parks, and residential landscapes in the southeastern United States, and management strategies for reducing its impact are needed. In this study, the systemic fungicide propiconazole completely inhibited mycelial growth of Raffaelea spp. in vitro at concentrations 0.1 parts per million (ppm) or greater and was fungitoxic at 1 ppm or greater, whereas the fungicide thiabendazole was less inhibitory. None of the ten mature redbay trees that received root-flare injections of propiconazole developed crown wilt symptoms for at least 30 weeks after being inoculated with Raffaelea spp., whereas nine of ten untreated control trees wilted in more than one-third of their crowns. Propiconazole was retained in the stem xylem for at least 7.5 months after injection but was more frequently detected in samples from trees injected 4.5 months earlier and was not well detected in small-diameter branches. Results suggest that propiconazole may be useful in preventing laurel wilt in redbay, but limitations and questions regarding duration of efficacy, rate of uptake, and efficacy under different levels of disease pressure remain.

First Report of Laurel Wilt Disease Caused by a Raffaelea sp. on Avocado in Florida.

Laurel wilt is a vascular disease of redbay (Persea borbonia (L.) Spreng.) and other plants in the family Lauraceae in the southeastern United States. It is caused by a fungus (Raffaelea sp.) that is vectored by a non-native insect of Asian origin, the redbay ambrosia beetle (Xyleborus glabratus Eichhoff) (1). Since the initial detection of the redbay ambrosia beetle near Savannah, GA in 2002, laurel wilt has caused widespread mortality of redbay in Georgia, South Carolina, and Florida (1). In September 2007, an avocado (Persea americana Mill.) tree planted approximately 10 years earlier in a residential neighborhood in Jacksonville, FL was discovered to be infected with laurel wilt. The crown was in various stages of decline, including upper branches that were dead and leafless, those with wilted and drooping foliage, and those with healthy foliage. Removal of bark from wilted branch sections revealed black-to-brown streaks of discoloration in the sapwood and a few ambrosia beetle holes from which the discoloration extended into the adjacent wood. A Raffaelea sp. was isolated from discolored wood samples by surface sterilizing wood chips by submersion in a 5% sodium hypochlorite solution for 30 s and plating them on cycloheximide streptomycin malt agar (2). Small subunit (18S) sequences from the rDNA were amplified by PCR and sequenced with primers NS1 and NS4 (3). BLASTn searches revealed homology to Raffaelea sp. C2203 (GenBank Accession No. EU123076, 100% similarity, e-value of 0.0, and a total score of 1,886), which is known to be the causal agent of laurel wilt (1). The small-subunit rDNA sequence for this isolate has been deposited into GenBank and has been assigned accession No. EU257806. Pathogenicity of the laurel wilt pathogen on Persea spp. in growth chamber trials has been previously demonstrated (1). Laurel wilt is of concern to the commercial avocado industry and is a potential threat to the Lauraceae elsewhere in the Americas.

Laurel Wilt: A Threat to Redbay, Avocado and Related Trees in Urban and Rural Landscapes

HS-1137, a 6-page illustrated fact sheet by Albert E. Mayfield III, Jonathan H. Crane, and Jason A. Smith, describes for homeowners this fungal disease of trees in the laurel family that is carried by the non-native redbay ambrosia beetle. Includes descriptions of the beetles, plant hosts, impact, host symptoms, and management strategies. Includes references. Published by the UF Horticultural Sciences Department, March 2008.