Evidence for the Widespread Occurrence of Bacteria Implicated in Acute Oak Decline from Incidental Genetic Sampling

Acute Oak Decline (AOD) is complex syndrome affecting Britain’s keystone native oak species, (Quercus robur L. and Q. petraea L. (Matt.) Liebl.), in some cases causing mortality within five years of symptom development. The most distinguishable symptom is weeping stem lesions, from which four species of bacteria have been isolated: Brenneria goodwinii, Gibbsiella quercinecans, Lonsdalea britannica and Rahnella victoriana. We do not yet know where else these bacteria exist, and little is known about the relationship of the wider oak leaf microbiome (phyllosphere) to acute oak decline. Here we investigate whether incidental evidence from a large oak genome re-sequencing dataset could be used to detect these bacteria in oak foliage, and whether bacterial incidence co-varied with AOD status or location. Oak leaves and buds were sampled from 421 trees at five sites in England. Whole genomic DNA from these samples was shot-gun sequenced with short reads. Non-oak reads were extracted from these data and queried to microbial databases. Reads uniquely matching AOD-associated bacterial genomes were found to be present on trees from all five sites and included trees with active lesions, trees with historic lesions and trees without AOD symptoms. The abundance of the AOD-associated bacteria did not differ between tree health categories but did differ among sites. We conclude that the AOD-associated bacteria may be members of the normal oak microbiome, whose presence on a tree is not sufficient to cause AOD symptoms.

Evidence for the Widespread Occurrence of Bacteria Implicated in Acute Oak Decline from Incidental Genetic Sampling

Acute Oak Decline (AOD) is complex syndrome affecting Britain’s keystone native oak species, (Quercus robur L. and Q. petraea L. (Matt.) Liebl.), in some cases causing mortality within five years of symptom development. The most distinguishable symptom is weeping stem lesions, from which four species of bacteria have been isolated: Brenneria goodwinii, Gibbsiella quercinecans, Lonsdalea britannica and Rahnella victoriana. We do not yet know where else these bacteria exist, and little is known about the relationship of the wider oak leaf microbiome (phyllosphere) to acute oak decline. Here we investigate whether incidental evidence from a large oak genome re-sequencing dataset could be used to detect these bacteria in oak foliage, and whether bacterial incidence co-varied with AOD status or location. Oak leaves and buds were sampled from 421 trees at five sites in England. Whole genomic DNA from these samples was shot-gun sequenced with short reads. Non-oak reads were extracted from these data and queried to microbial databases. Reads uniquely matching AOD-associated bacterial genomes were found to be present on trees from all five sites and included trees with active lesions, trees with historic lesions and trees without AOD symptoms. The abundance of the AOD-associated bacteria did not differ between tree health categories but did differ among sites. We conclude that the AOD-associated bacteria may be members of the normal oak microbiome, whose presence on a tree is not sufficient to cause AOD symptoms.

The distribution of bacteria Gibbsiella quercinecans and Brenneria goodwinii in oak (Quercus robur L.) stands in Latvia

Acute Oak Decline (AOD) is a complex plant disease affecting species of native oaks, for example, Quercus robur, in Latvia. Typical AOD symptoms are bark cracks with dark exudate. In other European region countries, like the United Kingdom, AOD is associated with bark insects, for example, Agrilus biguttatus. From the results of the forest survey, it can be concluded that in oak forest stands located in Latvia, there is practically no damage by the bark insect A. biguttatus, which may indicate the existence of another AOD spreading vector. In 2019, from one oak, which was positive for AOD in the spring, repeated stem bleed samples were taken in the autumn period, the laboratory analysis results of the repeated samplings were negative. These findings suggest that the activity of bacteria Brenneria goodwinii and Gibbsiella quercinecans are influenced by climatic conditions.

First report of Brenneria goodwinii and Gibbsiella quercinecans bacteria, detected on weaken oak trees in Poland

The decline of oak stands is a phenomenon that has been observed since the beginning of the 20th century in many European countries. It can be caused both with abiotic factors such as drought and fluctuations in groundwater levels, as well as biotic factors such as infestations by insects, fungi and bacteria. Acute Oak Decline (AOD) is an dangerous disease of oak trees which was first observed in the XX century. From the moment the first symptoms were noticed, the disease is able to kill trees in up to 6 years. Two species of bacteria, Brebberia goodwinii and Gibsiella quercinecand, are considered as one of the infectious agents of AOD. The decline of Quercs robur was reported from a weakened stand from the Chojnów Forest District. Symptomatic exudates were sampled and subjected to laboratory analysis. Using the Real time PCR, bacteria Brenneria goodwinii and Gibsiella quercinecans were confirmed in two out of seven collected field samples. This report is the first information on the observation of these bacteria in weakened stands with the participation of Quercus robur in Poland. Keywords: Quercus robur, bacteria, Brenneria goodwinii, Gibsiella quercinecans, real time PCR, Acute Oak Decline

Raffaelea quercina sp. nov. Associated with Cork Oak (Quercus suber L.) Decline in Portugal

Research Highlights: Raffaelea quercina sp. nov. is an ophiostomatoid fungus isolated from the ambrosia beetle Platypus cylindrus. The species occurs in symptomatic Portuguese cork oak trees, (Quercus suber L.), exhibiting vegetative decline. Background and Objectives: Quercus suber L. is a species restricted to the Mediterranean basin, of special economic importance as it constitutes the crucial raw material for the cork production industry, in particular for Portugal, the world’s leading producer. Over the last three decades a progressive and alarming decline of cork oak trees has been observed across its distribution area, including Portugal. The ambrosia beetle Platypus cylindrus, commonly known as the oak pinhole borer, establishes symbiotic relationships with fungi from which it depends for survival and for oak colonization. Some of these fungi are ophiostomatoid species of the Raffaelea genus, known as ambrosia fungi associated with ambrosia beetles. Some Raffaelea species exhibit phytopathogenic activity causing wilting and/or death of trees. The objective of the present study is to identify the association between P. cylindrus and Raffaelea species in Portuguese cork oak stands showing symptoms of disease and decline. Materials and Methods: A total of 300 adult insects were collected as they emerged from cork oak logs, sampled from symptomatic trees. Axenic isolates of Raffaelea species were obtained from the beetles and their galleries in the trunks and identified based on morphological features and molecular analysis of the SSU and LSU rDNA regions. Results: Two Raffaelea species were identified, i.e., R. montetyi and a novel Raffaelea species closely related to R. canadensis. The novel species is morphologically and genetically characterized in this study, and erected as Raffaelea quercina M.L. Inácio, E. Sousa & F. Nóbrega, sp. nov. Raffaelea quercina constitutes a new phytopathogenic fungal species associated with P. cylindrus and cork oak trees exhibiting symptoms of vegetative decline. Conclusions: Raffaelea species appear to have a significant role in cork oak decline. Future research on the association between P. cylindrus and Raffaelea species, encompassing the trans-European and North African wide-range of cork oak stands, would further clarify the relationships between ambrosia beetles, associated fungi and cork oak decline, contributing to a better understanding of the phenomena and for strategies aiming to halt the continuous decline of the unique cork oak stands enclosed in the Mediterranean basin.