- Author: Sara Garcia Figuera, Jennifer Reed and Brianna McGuire
Western Plant Protection Network at UC Davis
Early detection technologies (EDTs) are tests that indicate the presence of disease before signs or symptoms of the disease can be seen. In the same way that a doc-tor measures a patient's blood pressure to look for heart problems, a grower might use a trained “sniffer” dog to detect changes in a tree that looks healthy but has huanglongbing (HLB) disease. By using the EDT, the grower is able to uncover HLB earlier, and can decide on an early, cost-saving course of action.
In the case of HLB, there are many EDTs under development. Some of them look for patterns in the microorganisms that live on the citrus leaves (Leveau snapshot); some look for patterns in the chemicals that are produced by the tree in response to HLB (Pourreza, Davis and Slupsky); and others look for the molecules that the bacterium injects in the tree to cause disease (Ma). A description of some of these EDTs can be found on the Science for Citrus Health website.
Why do we need EDTs for HLB?
To understand why EDTs are needed and what their potential value is, it is necessary to understand the difference between the incubation period for a disease and the latent period. The incubation period is the time between exposure to the pathogen and the appearance of symptoms. The latent period is the time between exposure and the newly-infected host becoming infectious. Huanglongbing (HLB) has a long incubation period and a very short latent period, which means that a tree can be dis-eased for a long time without showing any visible symptoms, while being infectious for a large fraction of that time. Even if a tree does not seem diseased, it can serve as a home for the bacterium (Candidatus Liberibacter asiaticus, CLas) that causes HLB. If a psyllid feeds on the infected tissue of a tree (with or without symptoms), CLas that is present in the leaf tissue can be picked up by the insect and transmitted to other trees when the psyllid moves on to feed. Information from an EDT can help a grower detect the disease in a tree a long time before it would be detected by eye. This cuts down the time psyllids are able to feed on it and transmit the disease, slowing the spread of HLB to neighboring trees.
Why is it important to remove infected trees as early as possible?
If a tree that tests positive for CLas is not treated or removed, the bacterium will spread throughout the tree. Over time, an increasing proportion of the tree's tissues will become infected, increasing the chances that a psyllid will become infected upon feeding, and subsequently spread the infection to healthy neighboring trees. If the infected tree is removed, there is no opportunity for psyllids to feed on the infected tissue and spread the disease. Once CLas is detected, tree removal is the only surefire way to prevent the spread of the infection, and it is extremely time-sensitive. The sooner an infected tree is removed, the lower the chances that psyllids will get infected. The savings associated with early infected tree removal will be proportional to the amount of surrounding trees that would have been infected with CLas due to that tree, and the number of months that it would be left on the ground.
Who is working on the project?
Several research teams in different universities and research stations, supported by a variety of funding organizations, have been working on the development of a variety of EDTs. These EDTs, designed under laboratory and greenhouse conditions, are being validated under field conditions in Texas and Florida. In California, where HLB has not been detected in citrus orchards, samples of different citrus varieties have been collected from healthy trees and trees affected by other diseases from all over the state. These samples are being used to calibrate the EDTs, and to test if they can distinguish between healthy and HLB-diseased trees, and between HLB-diseased trees and trees affected by other common citrus diseases. Dr. Neil McRoberts and his team at UC Davis are evaluating the data from these experiments and providing support to the EDT researchers.
What are the challenges and opportunities?
Currently, regulations require HLB infected trees to be removed if a certain amount of CLas DNA is detected in leaf samples through polymerase chain reaction (PCR). However, CLas is unevenly distributed in the sap of citrus trees, and the leaf samples collected might not be PCR-positive even though the bacterium is already present elsewhere in the tree. EDTs offer the possibility to detect infected trees before they are PCR-positive, so they could be removed earlier in the HLB epidemic. Therefore, the value of EDTs relies on the voluntary removal of EDT-positive trees before the law requires them to be removed.
No EDT gives perfect diagnostic results. Sometimes healthy trees will produce EDT scores that look like diseased trees (so-called “false positives”). Removing such trees will result in an immediate financial loss. However, because the economic damage caused by leaving an infected tree in place is much bigger than the value of a healthy tree, using an EDT to guide decisions has the potential to result in a long-term economic benefit to individual growers and communities, by reducing the spread of HLB. Losing a few healthy trees along the way is the unavoidable cost of stopping the disease from spreading. Like-wise, some trees will seem healthy based on EDT scores but might end up showing symptoms (“false negatives”). The proportion of true positives, false positives, true negatives and false negatives represents the accuracy of a diagnostic test. Dr. McRoberts' team is analyzing the accuracy of the EDTs, and preliminary results suggest that the best performing EDTs could be correctly determining the status of the trees 95% of the time.
The results of this analysis could be used to foster the adoption of EDTs among the citrus grower community, promoting the idea that the sooner infected trees are detected and removed, the smaller impact HLB will have on California's citrus production. Unless there is sufficient cooperation in integrated management of HLB by removing infected trees as early as possible, controlling the ACP on an area-wide scale, and using certified plant material, the California citrus industry is likely to suffer un-sustainable economic losses to HLB.
- Author: Joey Mayorquin, Mohamed Nouris, Akif Eskalen and Florent Trouillas
Figure 1: Citrus shoot dieback (top) and gummosis (bottom) caused by Colletrotrichum.
A new disease of citrus has been found in the main growing regions of the Central Valley of California. The causal agents of this disease were identified as species of Colletotrichum, which are well-known pathogens of citrus and other crops causing anthracnose diseases. Several growers and nurserymen in various orchards in the Central Valley first noticed the disease in 2013. Symptoms include leaf chlorosis, crown thinning, gumming on twigs and shoots dieback, and in severe cases, death of young trees. The most characteristic symptoms of this disease are the gum pockets, which appear on young shoots either alone or in clusters and the dieback of twigs and shoots (Fig.1). Field observations indicate that symptoms initially appear during the early summer months and continue to express until the early fall. These symptoms were primarily reported from clementine, mandarin, and navel orange varieties. In order to determine the main cause of this disease, field surveys were conducted in several orchards throughout the Central Valley. Isolations from symptomatic plant samples frequently yielded Colletotrichum species. Morphological and molecular phylogenetic studies allowed the identification of two distinct species of Colletotrichum (Colletotrichum karstii and Colletotrichum gloeosporioides) associated with twig and shoot dieback. Interestingly, these Colletotrichum species were also isolated from cankers in larger branches. Although C. gloeosporioides is known to cause anthracnose on citrus, a post-harvest disease causing fruit decay, it has not been reported to cause shoot dieback of citrus. C. karstii however has not been reported previously from citrus in California and our research team is currently conducting field and green house studies to determine the pathogenicity of this species in citrus. At present, it is unclear how widespread this disease is in California orchards or how many citrus varieties are susceptible to this disease. Pest control advisors are monitoring citrus trees for the presence of the disease in the Central Valley (particularly clementine, mandarin, and navel varieties) during the early summer months. Continuing research led by Dr. Akif Eskalen in collaboration with Dr. Florent Trouillas is focused on further understanding the biology of the fungal pathogens as well as factors influencing disease expression in order to develop management strategies against this emerging disease.
The Citrus Research and Development Foundation (CRDF), a $124 million state citrus-industry initiative, has invested nearly 90 percent of its funds in HLB research. CRDF asked the Academies to review its research portfolio and determine if its efforts have followed recommendations outlined in the Academies' 2010 report, which originally called for the organization's creation. The committee found that CRDF was responsive to several recommendations from the previous report, and along with other funders, has advanced our knowledge about the disease. However, HLB remains a serious danger to Florida's citrus industry, having progressed from an acute to a chronic disease throughout the state.
The report notes that significant barriers to progress toward an HLB solution still exist, among them the inability to culture the bacteria in the laboratory, the lack of advanced diagnostics for early disease detection, and the absence of standardized research methodology that would improve the comparability of results across studies. Resolution of any one of these issues would constitute a significant step, according to the report.
The committee recommended continuing support for both basic and applied research for short- and long-term research efforts. In the long run, HLB solutions would likely utilize new technology, such as gene modification and gene editing, focusing on targets that mediate molecular interactions among plant, bacteria, and the vector, the committee said. As interest in using genetic modification in research grows, CRDF should also consider funding research to assess stakeholder acceptance of the technology and expand efforts to educate growers, processors, and consumers to facilitate the eventual deployment of genetically modified citrus lines.
In the meantime, growers in the state will need short-term solutions for the industry to remain viable. The report recommends finding the best suite of strategies to control the disease in different environmental and growing conditions, vector and pathogen pressures, tree varieties, and stages of tree health, which would help growers in Florida and other states where HLB also occurs.
The report also highlights the need to better understand the economic and sociological factors that impact decision-making and behaviors of growers, which influence the adoption of HLB management strategies. CRDF should create accessible databases to support sociological and economic modeling of citrus greening-related research outcomes and application projections.
The report recommends researchers communicate about the outcomes and evaluation of their efforts in a timely and systematic way. Additionally, current approaches to research prioritization and funding based within individual federal and state funding agencies have not led to development of a master plan for HLB research and subsequent management solutions. CRDF should work with other funding agencies to create an overarching advisory panel to develop a master plan for HLB research, communication, and management.
The study was sponsored by CRDF. The National Academies of Sciences, Engineering, and Medicine are private, nonprofit institutions that provide independent, objective analysis and advice to the nation to solve complex problems and inform public policy decisions related to science, technology, and medicine. The National Academies operate under an 1863 congressional charter to the National Academy of Sciences, signed by President Lincoln. For more information, visit http://national-academies.org. A roster follows.
To download full report: https://www.nap.edu/read/25026/chapter/1