Source: TEXAS A&M UNIVERSITY submitted to NRP
PROTECTING TREE HEALTH IN TEXAS COMMUNITIES AND WOODLANDS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1017595
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Oct 3, 2018
Project End Date
Oct 3, 2023
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
TEXAS A&M UNIVERSITY
750 AGRONOMY RD STE 2701
COLLEGE STATION,TX 77843-0001
Performing Department
Plant Pathology & Microbiology
Non Technical Summary
Our valuable natural forests and urban trees are under a barrage of emerging and introduced tree pathogens. This is particularly true in Texas where the enormous value of these resources are subject to extremely stressful climates and a continual invasion of exotictree pathogens acrosslengthy borders. Solutions to the diseases areas complex as the pathogens themselves, the tree hosts they infect and the diseases they cause. There is relatively little research currently being conducted to provide practical, effective options for directly managing tree pathogens and mitigating thier impacts. This is particularly obvious when interacting with clientele expecting some useful response from AgriLife Extension when they reach out for solutions to their tree problems in Texas. Improvements in early detection and diagnostics would be very helpful, because these issues are the first line ofdefense against any plant pathogen. For improving plant disease diagnosis, studies to develop a RT-PCR protocol for detecting the oak wilt pathogen, Bretziella fagacearum will be conducted in this proposed M-S project. There are several limitations to the most common method for diagnosing a tree suspected of being infected with oak wilt. This method relies on growing the fungus out of the sample in the laboratory, and can take weeks to complete and may have unacceptably high levels of false negative results (the tree may be infected but the sample does not contain the fungus). An RT-PCR method will be quicker and far more sensitive than isolating the pathogen, but requires a great deal of effort to verify the reliability and consistency of the test. Another potential advantage would the ability to successfully conduct a post mortum on diseased trees to diagnose a case of oak wilt after it the tree has died.The other issue, concerning the monitioring of exotic pathogens, will also be a focus of the proposed project There will be surveys for Citrus Greening, caused by a bacterium called Candidatus liberabacter asiaticus, and Sudden Oak Death, caused by the fungus Phytophthora ramorum. These two pathogens pose an immediate and significant threat to tthe Texas citrus industry as well as the considerable oak resources of the State. Another important issue that will be addressed is the need for improvements to methods ofdirect control and saving trees at threat of infection by the oak wilt pathogen. Tree injection with a fungicide has proven to be an effective option in the management of oak wilt. One technique, called the macro-injection method, has dominated the injection market for the past 20 years because there were reliable research results to support its use. More recently, there has been a proliferation of new methods for injecting trees, mostly based on "micro" injection methods. These new methods are purportedly faster, less laborius, and better for the tree than macroinjection. However, the only evidence to support these claims is in need of further verification and testing. That is another objective of this project, to test methods of injecting trees with both systems under conditions of natural infection by the pathogen to determine whether the advantages and disadvantages to how trees are currently being injected. The information generated by the proposed studies will be of great interest to a wide variety of clientele in both the public and private sectors. One of the greatest impacts will be on the Central Texas urban forests where there is a thriving, well - trained and organized arborilogical industry in Texas ready to adopt better technology for dealing with oak wilt. The PI regularly meets with arborists and homeowners at 25 - 30 meetings annually, including several Master Gardener trainings in Texas counties. Factsheets will be published through the Texas A&M AgriLIfe Bookstore for distribution on the internet, and there will publications in peer-reveiwed journals for the braoder scientific community.
Animal Health Component
80%
Research Effort Categories
Basic
10%
Applied
80%
Developmental
10%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2120699116050%
1242110116050%
Goals / Objectives
The value of trees to the State of Texas can be expressed in many ways. The total economic output of the East Texas forest timber industry in 2015 was $29,849,220,000. Timber alone was ranked ninth among Texas' top ag commodities and forest based industry was one of the top 10 manufacturing sectors in the state (see website http://txforestservice.tamu.edu/main/default.aspx). Although more difficult to quantify, the urban forest values in the state are also considerable. For example, a recent study by the U.S. Forest Service revealed that the compensatory value of trees in Austin, TX, in terms of air pollution removal, reduced carbon emissions, carbon storage and sequestration, reduced energy use for buildings, storm water capture, etc., was $16 billion. Similar staggering values can be found for Houston, San Antonio, Dallas, and other metropolitan areas throughout the state. However, economic values do not tell the whole story as far as the value trees have to the state in terms of value of life and ecosystem services. For example, live oak (Quercus virginiana, Q. fusiformis) is represented on the original state seal of Texas, going back to 1845, as a symbol of strength and endurance. This connection with live oak was most recently exemplified by the national recoginition of the iconic Goose Island Oak near Rockport, TX, when it survived a direct strike by Hurricane Harvey. The trees of Texas are economic and environmental assets that justify the effort to better protect them from the ravages of pathogenic agents.A wide variety of forest types and woodlands occurs in Texas (Correll and Johnston 1970, also see website of the Bureau of Economic Geology, University of Texas http://www.beg.utexas.edu/UTopia/images/pagesizemaps/vegetation.pdf). There are therefore many different species of trees, each with its complement of diseases. Some diseases are caused by existing, infectious pathogens with the potentials to cause extremely destructive epidemics, such as oak wilt and Dutch elm disease. Other diseases appear to be less pathogenic, but are increasing in incidence and severity with changes in climatic conditions (Kliejunas et al. 2009). Also, the introduction of exotic, invasive pathogens on trees is an ongoing threat in Texas, with the potential for a huge impact on valuable landscape and forest trees (Tkacz et al. 1998, Hulcr and Dunn 2011).Monitoring and studying tree diseases, regardless of their origins, are warranted when they threaten the extremely valuable Texas woodlands, forests, and urban landscapes. This project addresses those threats with better detection, improved understanding of their epidemiology, and the ultimate goal of reducing their impact. New and refined disease prevention and control methods resulting from these studies will be made available through educational outreach activities to the vast clientele of the Texas A&M AgriLife Extension Service. These research goals are consistent with the emerging knowledge areas, as defined in the 2010 McIntire-Stennis Strategic Plan update. Specifically, in the first of two recommendations, pathogens are specifically cited in the Foundational Areas of Knowledge as a focus of research. A fundamental understanding of pathogen function is ".....critical to achieving a broader knowledge of forests, watersheds, and global functions." In addition, technological advancements are needed in areas such as nanotechnology, remote sensing, and biotetchnology in order to monitor, assess, and expand the productivity and utility of forests. The advancement and application of new state-of-the-art technologies will advance our capacity to protect forests.Given the high value of trees as a critical natural resource, and given the existing disease threats to that resource, the major goals of this project are to: I. Identify and assess the disease threats to Texas forests, woodlands, and urban forests of Texas II.Improve existing methods to control or manage disease threates to Texas forests, woodlands, and urban forests. III. Disseminate project activities and results to relevant audiences.
Project Methods
ProceduresObjective I. Identify and assess the disease threats to Texas forests, woodlands, and urban forests of Texas. Approach. The surveillance of tree diseases can be accomplished through a number of means. Given the need to cover such a large area, a hierarchical approach must be used to gather data at different scales. County Extension Agents with Texas AgriLife Extension and their volunteers, such as the Master Gardeners and Master Naturalists, will be trained to assist with statewide monitoring for new and emerging pathogens. Training of arborists, landscape professionals, nurserymen, and natural resource managers through workshops, websites (i.e., www.texasinvasives.org), and various educational outputs will also be an effective strategy to alert practitioners of threats. At a finer scale, there are prescribed survey protocols for detection of exotic pathogens with suspected routes of entry. In the case of SOD, surveys will focus on locations where the pathogen was previously found. Off-site, streamside surveys will consist of baiting waterways downstream from the sites and processing the baits for detecting P. ramorum in the lab (see the Forest Health Monitoring website http://www.fhm.fs.fed.us/sp/sod/sod.shtml). In addition, the same locations will undergo on-site soil and water sampling (see website of USDA APHIS PPQ website http://www.aphis.usda.gov/plant_health/plant_pest_info/pram/ for the latest protocols). For the detection of citrus greening, the focus will be on the Asian citrus psyllid, Diophorina citri Kuwayama, since the pathogen has yet to be found in Texas. Dooryard and landscape citrus trees will be sampled for psyllids and suspect tissues exhibiting symptoms of greening. Appropriate sampling locations will be provided by Master Gardeners, Master Naturalists, County Extension Agents, and urban foresters. These samples will be returned to the Texas Plant Disease Diagnostic Laboratory for detection of C. l. asiaticus using prescribed molecular protocols (see website of the USDA APHIS http://www.aphis.usda.gov/plant_health/plant_pest_info/citrus_greening/index.shtml). Citrus canker is another introduced bacterium with the potential for economic impact on the the greater commercial citrus industry in South Texas, and must be monitored throughout the Upper Gul Coast dooryard citrus population (Gottwald, T.R. 2000. Citrus canker. The Plant Health Instructor. DOI: 10.1094/PHI-I-2000-1002-01).Objective II. Improve existing methods to control or manage disease threats to Texas forests, woodlands, and urban forests.Part 1. Test and adopt new tools to diagnose oak wilt in a clinical setting. Approach.Each phase in the diagnostic process using qPCR to detect Bretziellla fagacearum (formally known as Ceratisistis fagacerarum) for clinical testing will be examined. Sapwood samples will be collected from infected and non-infected live oaks (Quercus fusiformis) in Central Texas for detection utilizing the standard culturing technique and comparing those results to testing with qPCR. The samples will be processed in the Texas Plant Disease Diagnostic Laboratory (TPDDL), Texas A&M AgriLife Extension Service in College Station, TX. The samples will be extracted using various extraction methods and specific probes and primers will be used for qPCR with an Applied Biosystems® 7500 Real-Time PCR System (Applied Biosystems Inc., Foster City, CA). Sample collection will vary according to season, Quercus sp., degree of symptom development in diseased trees, geographic location, sample retrieval and handling, and other factors that might vary during the course of submission to the TPDDL.Part 2. Test new injection protocols for treating high risk oaks threatened by infection by the oak wilt pathogen. Approach. The study is being conducted in central Texas within untreated, expanding oak wilt centers on privately-owned property within the range of live oak and oak wilt in central Texas. Non-symptomatic test trees (~125), ranging from 15 to 46 cm (6 - 18 in) dbh (diameter at breast height), will be selected in proximity with trees showing oak wilt symptoms (veinal necrosis). In April 2016, fifty (50) trees per delivery system (10 trees per site in five different sites) were injected with Alamo® (Syngenta) at the label rate (20 ml/inch tree dbh) using the two systems described above. A minimum of twenty-five (25) trees (5 trees per study site) serve as untreated controls. The application procedure used to inject the propiconazole formulation is based on the recommendations of each system manufacturer. All of the injected or check trees are located adjacent to the infected trees, at a distance of at least 50 - 75 ft but exhibiting no symptoms of oak wilt infection. The treatments are therefore being tested under conditions of natural infection with the pathogen. Trees have been evaluated for oak wilt symptoms at 6 month intervals since the treated. Each tree crown is given a rating of 0 (healthy), 1 (wilt symptoms comprising up to one-third of the crown), 2 (wilt symptoms comprising greater than one-third of the crown) (Mayfield et al. 2008), or 3 (dead tree). At each rating period, trees with a crown rating of 2 may be sampled from the stem and branches to determine the presence of Ceratocystis fagacearum. At the termination of the experiment in June 2019, final crown ratings will be made. An analysis of variance will be used to test for differences among injection systems. A X2 (Chi-square) test for homogeneity will be used to test the null hypothesis that the percentage of trees with a crown rating of 2 did not differ between the fungicide-treated trees and the untreated control group (Mayfield et al. 2008). The null hypothesis will be rejected if more than 20% of the fungicide-treated trees reached a crown rating of 2. The test will be invalidated if fewer than 60% of the control trees reach a crown rating of 2.Objective III. Disseminate project activities and results to all relevant audiences throughout Texas. Approach. The target audience(s) correspondwith each of the outcomes of this project. Information generated from the project is disseminated through many diverse county programs usually involving homeowners and landowners. The PI usually has direct contacts with 1,000 to 1,500 participants in these county programs, sponsored by the Texas A&M AgriLife County Extension Agents. Other audiences includenatural resource managers, ranch managers, and master gardeners. All of these groups are trainers and serve to deliver high quality, technically based information on the identification, biology and control of tree diseases. The groups are reached through a variety of trainings, workshops, and invited presentations sponsored by TAMU AgriLife, the Texas Forest Service, the Texas Chapter of the International Society of Arboriculture, and other entities. The PI also has a teaching appointment and is responsible for two courses where the information generated by this project are presented. These include ESSM 307 Forest Protection, and PLPA 301 Plant Pathology.

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:The primary target for the results of this project are landowners and homeowners in Texas. My efforts are directed at developing methods to prevent trees from becoming diseased and to treat them when they do. In order to reach the primary audience effectively, it is also necessary to extend the results to natural resource managers, county extension agents, commercial arborists, landscape managers and master gardeners with the best information available to help people with their tree problems. Validation and review of the results are accomplished through comunication with research scientists and extension specialists in other states and colleagues in relevant professional societies. Further evidence of the effectiveness of these efforts are found in participant assessments conducted by County Extension Agents following workshops and presentations made in support of county programs. The methods and results utilized in the project are relevant to the curricula of undergraduate and graduate students in courses on forest protection, methods in plant pathology., and invasive species. Changes/Problems:There will be no major changes in the problems being investigated for the coming year. However, there will be significant differences in the manner in which extension outreach will be carried out. Few face-to-face venues will be hosted by the Texas Forest Service and TAMU AgriLife Extension Service in 2021 in order to comply with ongoing efforts to reduce spread of COVID-19. As a result, more training videos and on line meetings will be conducted with clientele. There will also be an emphasis on the production of fact sheets and other popular publications to reach our audience. What opportunities for training and professional development has the project provided?Oak wilt workshops are routinely scheduled by the Texas A&M Texas Forest Service and theTexas A&M AgriLife Extension Service. Many of these workshops provide CEUs to arborists and other landscapes professionals needing compliance with Texas DEpartment of Agriculture licensing requirments. A previousUS Forest Service sponosored injection study provided the information to greatly enhance workshops where injection is demonstrated. For example, three "Oak Wilt Qualification" workshops are held every summer by the Texas Chapter of the International Society of Arboriculture in cooperation with the Texas A&M Texas Forest Service and Texas A&M AgriLife Extension Service. We now have evidence for communicating the relative efficacies of two of the most widely used injection methods, the macro system and the hybrid system. The information derived from the injection study also provides homeowners much needed information when deciding what injection approach they sould prefer when hiring arborists to treat their trees. During this reporting period, I also participated in two, 2 day workshops hosted by the Texas Chapter of ISA on tree diagnostics. One of these workshops was a hands on laboratory experience for a small group (n = 25). The other was a Tree Academy offered on line in which over 400 participants attended for 6 hours of training in tree diagnostics. Finally, a workshop was held for Texas A&M AgriLife County Extesnion Agents in the West Region on tree disease problems and solutions. How have the results been disseminated to communities of interest?As the the PI on this project, I have a "three way" appoinmtment, with responsibilities for teaching, research and extension in the TAMU System. This provides the opportunity to disseminate information on tree health issues to a very broad, diverse audience using a number of methods to get the information to users. Nearly 150 undergraduate and 10 graduate students enroll in courses I teach dealing with invasive species, tree disease control and identification, plant diagnositcs, and introductory plant pathology. These topics relate directly to the objectives of the various surveys and research projects that are in my project. I participate in 25-30 workshops through county extension, professional societies, agnecy trainings, and other venues, reaching thousands of attendees every year. I am very active in creating vidoes for training purposes in cooperation with the Texas Forest Service, where they exist on the TFS Oak Wilt Partnership website (texasoakwilt.org). I also help produce informational videos in cooperation with the Texas Plant Disease Diagnostic Laboratory called "What Wednesday". These short videos address topical tree disease problems that are submitted to the clinic for diagnostics. I participate in 3 oak wilt qualification trainings annually to certify commercial arborists in the diagnosis and management of the disease, resulting in 75 arborists receiving advanced training and the ability to have their companies listed on texasoakwilt.org. These intensive trainings are two days long, during which there is time to update attendees on the latest research and survey results from my lab. The quality and depth of information we are able to provide to our clientele is greatly enhanced by the continual production of new and relevant information from this project. What do you plan to do during the next reporting period to accomplish the goals?Given the limited opportunities for face-to-face education, a greater emphasis will be placed on the production of videos and factsheets to enhance my outreach efforts.

Impacts
What was accomplished under these goals? Two tree disease surveys were conducted during the reporting period. These surveys are intended to expand our knowledge of the potential spread of three very different pathogens; Phytophthora ramorum, the causal agent of sudden oak death, Candidatus Liberabacter asiaticus, the causal agent of citrus greening, and Xanthmonas axanopodis, the causal agent of citrus bacterial canker. The SOD survey is sponsored by the USDA Forest Service and focuses on sampling for P. ramorum in in waterways near nurseries where suspect plants were shipped. Five sites were sampled on 7 dates during the reporting period, including 2 sites in the fall season and 3 in the spring. Sampling was restricted during the reporting period due to travel restrictions in place to deal with the COVID pandemic. In no case was P. ramorum detected. This includes two of the nursery sites previously found to be positive in 2013 for P. ramorum, MAAS nursery in Seabrook, TX, and Paradise Palm nursery, in Webster, TX. There is little likelihood the pathogen is surviving at these locations. Streamside surveys combined with soil surveys carried out within the nurseries have been conducted annually since the detection, with no subsequent positive results. Both sites will be periodically sampled in the future, but we consider the threat of an outbreak of sudden oak death at those locations to be minimal. In cooperation with Dr. Awinash Bhatkar, Coordinator, Biosecurity and Agriculture Resource Management, Texas Dept. of Agriculture, our lab is periodically updated on the shipment of potentialP. ramorum-contaminated nursery stock into Texas wholesale and retail nursery operations. The three nurseries sampled in the Spring, 2020, were selected on this basis. This is the first time they were sampled, and plans are to sample again in future efforts. For citrus greening, the survey consists of two stages: sample collection and laboratory analyses. Samples consist of Asian Citrus Pyllids and suspect foliage from symptomatic citrus. These samples are returned to the Texas Plant Disease Diagnostic Laboratory (TPDDL) for laboratory diagnostics. In the case of citrus canker, trees are inspected for the presence of the typical symptoms of the disease at the same sites as those surveyed for citrus greening. Most prominent among those symptoms are raised, pustular lesions with a corky surface that form mostly on the lower leaf surface. Field diagnosis is aided by the presence of the yellow halo that surrounds thele sions. Similar lesions may be found on the fruit and (http://www.apsnet.org/edcenter/intropp/lessons/prokaryotes/Pages/CitrusCanker.aspx) stems of infected trees. If suspicious foliage, fruit, or branch tissues are observed, samples will be collected for testing at the TPDDL. For the current year, four counties were included in the survey (Chambers, Jefferson, Montgomery, and Orange). Due to the COVID-19 pandemic, the sampling opportunities were greatly diminished. Homeowners were reluctant to host survey crews on their properties during the spring and summer of 2020, so sample numbers were low. None of the collected samples were positive for either pathogen. One further study was initiated during the reporting period on oak wilt control. This studey involves the concern for resistance to the fungicide propiconazole developing in the Bretziella fagacearum population in Texas. Hundreds of thousands of trees (or more) have been treated with propiconazole since it was first labelled for use in the early 1990s. Hundreds of trees are typically treated repeatedly for limiting the growth of the pathogen in the trees and their root systems. This constant exposure of the pathogen population to the triazole fungicide could put pressure on the fungus to develop resistance, as other crop pathogens have. TwelveB. fagacearum isolates have been collected and are being analyzed for variability in growth and spore germination in poison agar assays. Results from this study will be analyzed and discussed in future reports.

Publications

  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Alabi, O.J., McBride, S., Appel, D.N., Al Rwahnih, M., and F. Pontasch. 2020. Complete genome sequence analysis of a grapevine cv. Blanc du Bois genetic variant of grapevine virus L. Arch. Virol. 165:1905-1909.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Appel, D.N., McBride, S., Davis, R., And Alabi, O.J. 2020. The occurrence of mixed virus infections in a Texas Blanc du Bois vineyard. (https//doi.org/10.1094/PHTYO-110-12.S269)


Progress 10/03/18 to 09/30/19

Outputs
Target Audience:The primary target for the results of this project are landowners and homeowners in Texas. My efforts are directed atdevelopingmethods to prevent trees from becoming diseased and to treat them when they do. In order to reach the primary audienceeffectively, it is also necessary to extend the results tonatural resource managers, county extension agents, commercial arborists, landscape managers and master gardeners with the best information available to help people with their tree problems. Validation and review of the results are accomplished through coomunication with research scientists and extension specialists in other states and colleagues inrelevant professional societies. The methods and results utilized in the project are related to the curricula of undergraduate and graduate students in courses on forest protection, methods in plant pathology., and invasive species. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Tree injection is a popular, effective, and economically important tool in the management of oak wilt in Texas.Oak wiltworkshops are routinely scheduled by the Texas A&M Texas Forest Service andtheTexas A&M AgriLife Extension Service. The US Forest Service sponosored injection study provided the information to greatly enhance workshops where injection is demonstrated.For example, three "Oak Wilt Qualification" workshops are held every summer by the Texas Chapter of the International Society of Arboriculture in cooperation with the Texas A&M Texas Forest Service and Texas A&M AgriLifeExtension Service.We now have evidence for communicating the relative efficacies of two of the most widely used injection methods, the macro system and the hybrid system.The information derived from the injection study also provides homeowners much needed information when deciding what injection approach they sould prefer when hiring arboriststo treat their trees. How have the results been disseminated to communities of interest?As the the PI on this project, I have a "three way" appointment, with responsibilities for teaching, research and extension in the TAMU System. This provides the opportunity to disseminate information on tree health issues to a very broad, diverse audience using a number of methods to get the information to users.Nearly 150 undergraduate and 10 graduate students enroll in courses I teach dealing with invasive species, tree disease control and identification, plant diagnositcs, and introductory plant pathology.These topics relate directly tothe objectives of the various surveys and research projects that are in my project. Iparticipate in 25-30 workshops through county extension, professional societies, agnecy trainings, and other venues, reaching thousands of attendees every year. I am very active in creating vidoes for training purposes in cooperation with the Texas Forest Service, where they exist on the TFS Oak Wilt Partnership website (texasoakwilt.org). I also help produce informational videos in cooperation with the Texas Plant Disease Diagnostic Laboratory called "What Wednesday". These short videos address topicaltree disease problems that are submitted to the clinic for diagnostics. I participate in 3 oak wilt qualification trainings annually to certify commercial arborists in the diagnosis and management of the disease, resulting in 75 arborists receiving advanced training and the ability to have their companies listed on texasoakwilt.org. These intensive trainings are two days long, during which there is time to update attendees on the latest research and survey results from my lab.Although no new factsheets were published through the Texas A&M AgriLife Extension Bookstore during the reporting period, 2 are in preparation and will be published during the coming year. The quality and depth of information we are able to provide to our clientele is greatly enhanced by the continual production of new and relevant information from this project. What do you plan to do during the next reporting period to accomplish the goals?With the conclusion of the injection study, a new direction is being taken to improve diagnostic capabilities for early detection and diagnosis of oak wilt. Some seed money has been acquired to improve on molecular tools previously developed under the auspices of this project during previous years. Survey activities will be continued for the coming year, as will outreach and teaching.

Impacts
What was accomplished under these goals? Two tree disease surveys were conducted during the reporting period. These surveys are intended to expand our knowledge of the potential spread of threevery different pathogens; Phytophthora ramorum, the causal agent of sudden oak death, CandidatusLiberabacter asiaticus, the causal agent of citrus greening, and Xanthmonas axanopodis, the causal agent of citrus bacterial canker. The SOD survey is sponsored by the USDA Forest Service and focuses on sampling for P. ramorum in in waterways near nurseries where suspect plants were shipped. Six siteswere sampled on 18 occasions during the reporting period, including 3 sites in the fall season and 3 in the spring. In no case was P. ramorum detected. This includes two of the nursery sites previously found to be positive in 2013 for P. ramorum, MAAS nursery in Seabrook, TX, and Paradise Palm nursery, in Webster, TX. There is little likelihood the pathogen is surviving at these locations. Streamside surveys combined with soil surveys carried out within the nurseries have been conducted annually since the detection, with no subsequent positive results. Both sites will be periodically sampled in the future, but we consider the threat of an outbreak of sudden oak death at those locations to be minimal. In cooperation with Dr. Awinash Bhatkar, Coordinator, Biosecurity and Agriculture Resource Management, Texas Dept. of Agriculture, our lab is periodically updated on the shipment of potentially P. ramorum-contaminated nursery stock into Texas wholesale and retail nursery operations. The three nurseries (TX70, TX71, and TX72, see Table 1) sampled in the Spring, 2019, were selected on this basis. This is the first time they were sampled, and plans are to sample again in future efforts. For citrus greening, the survey consists of two stages: sample collection and laboratory analyses. Samples consist of Asian Citrus Pyllids and suspect foliage from symptomatic citrus. These samples are returned to the Texas Plant Disease Diagnostic Laboratory (TPDDL) for laboratory diagnostics. In the case of citrus canker, trees are inspected for the presence of the typical symptoms of the disease at the same sites as those surveyed for citrus greening. Most prominent among those symptoms are raised, pustular lesions with a corky surface that form mostly on the lower leaf surface. Field diagnosis is aided by the presence of the yellow halo that surrounds the lesions. Similar lesions may be found on the fruit and (http://www.apsnet.org/edcenter/intropp/lessons/prokaryotes/Pages/CitrusCanker.aspx) stems of infected trees. If suspicious foliage, fruit, or branch tissues are observed, samples will be collected for testing at the TPDDL.For the current year,four counties were included in the survey (Chambers, Jefferson, Montgomery, and Orange). Due to changes in the counties in the citrus survey from previous years, the discovery of new, reliable sites took longer than anticipated. Chambers and Montgomery counties do not have the same high concentration of dooryard citrus as compared to other coastal counties. The emergence of psyllid populations in the survey area was delayed until July this year, probably due to the effects of some severe cold weather during the spring. This was first noted in the previous semi-annual report. As a result of freeze, most of the citrus trees were damaged thus reducing the available foliage for healthy flush to support the psyllids. A total of 72 site visits were made with 357 trees inspected (observations) (Table 1). No tree had symptoms of bacterial canker, and foliage from 38 trees were sampled to test for greening. None of them were positive, nor did any of the 16 psyllid samples test positive for greening. A three year study on improving tree injection methods for management of oak wilt, caused by Bretziella fagacearum, was concluded during the reporting period. Two tree injection methods were compared by treating naturally infected trees on a ranch with oak wilt in Pipe Creek, TX. The first injection method was a high volume, low concentration method called the "macro" system. The second was a "hybrid" system comprised of a relatively low vaolume, high concentration method. The most significant results of this study clearly demonstrate that the macro injection method and the hybrid injector are both similarly effective in preventing disease development in trees being treated before becoming infected by B. fagacearum. Traditionally, the macro injection system was so named for the relatively large holes, or injection ports, when compared to the smaller ports needed for the micro injectors. In addition, macro injection consists of high volumes of dilute fungicide rates relative to the low volume, high concentrations used with micro injectors. The hybrid technique uses high concentrations of fungicide, as do micro techniques, but the injection hole is larger than both the macro and micro technique. In addition, the ports can be connected in a series with the hybrid technique, in a manner similar to the macro method of injection. For these reasons, we have proposed to use the term hybrid to designate injection equipment using multiple, small containers, concentrated fungicide, relatively large injection holes, and ports connected in a series to distinguish them from the small, separtae capsules used in micro injection methods. Another valuable result from this study was the opportunity to document the relative advantages and disadvantages for the two injection techniques in an objective manner. For example, the hybrid injection method requires the use of a plug that is inserted into the hole of the tree immediately following drilling. The injection port is then inserted into a self-sealing diaphragm in the plug, which remains permanently in the tree following treatment. These plugs apparently delay the ability of the tree to compartmentalize and seal the wound site for the hybrid injectors, leading to considerable leakage. A visible crack is often seen above and below the injection site as well (Figure 4)l. This response does not occur with the macro system, and should be a source of concern, particularly for younger trees where decay could be a problem in later years.

Publications