he genome and genetics of a high oxidative stress tolerant Serratia sp. LCN16 isolated from the plant parasitic nematode Bursaphelenchus xylophilus

Background: Pine wilt disease (PWD) is a worldwide threat to pine forests, and is caused by the pine wood nematode (PWN) Bursaphelenchus xylophilus. Bacteria are known to be associated with PWN and may have an important role in PWD. Serratia sp. LCN16 is a PWN-associated bacterium, highly resistant to oxidative stress in vitro, and which beneficially contributes to the PWN survival under these conditions. Oxidative stress is generated as a part of the basal defense mechanism used by plants to combat pathogenic invasion. Here, we studied the biology of Serratia sp. LCN16 through genome analyses, and further investigated, using reverse genetics, the role of two genes directly involved in the neutralization of H2O2, namely the H2O2 transcriptional factor oxyR; and the H2O2-targeting enzyme, catalase katA. Results: Serratia sp. LCN16 is phylogenetically most closely related to the phytosphere group of Serratia, which includes S. proteamaculans, S. grimessi and S. liquefaciens. Likewise, Serratia sp. LCN16 shares many features with endophytes (plant-associated bacteria), such as genes coding for plant polymer degrading enzymes, iron uptake/ transport, siderophore and phytohormone synthesis, aromatic compound degradation and detoxification enzymes. OxyR and KatA are directly involved in the high tolerance to H2O2 of Serratia sp. LCN16. Under oxidative stress, Serratia sp. LCN16 expresses katA independently of OxyR in contrast with katG which is under positive regulation of OxyR. Serratia sp. LCN16 mutants for oxyR (oxyR::int(614)) and katA (katA::int(808)) were sensitive to H2O2 in relation with wild-type, and both failed to protect the PWN from H2O2-stress exposure. Moreover, both mutants showed different phenotypes in terms of biofilm production and swimming/swarming behaviors. Conclusions: This study provides new insights into the biology of PWN-associated bacteria Serratia sp. LCN16 and its extreme resistance to oxidative stress conditions, encouraging further research on the potential role of this bacterium in interaction with PWN in planta environment.

Natural recovery of the benthic nematode assemblages associated to Zostera noltii seagrass beds after physical disturbance caused by digging harvest activity

Seagrass beds are productive ecosystems that maintain high levels of biodiversity, making them susceptible to anthropogenic pressures such as bivalve harvesting. Nematodes are considered great ecological indicators as changes in their density, diversity and structure may represent changes in the environment. This experimental fieldwork aimed to assess the impact of the bivalve harvesting on the nematodes assemblage of a seagrass bed in the Mira estuary by simulating the digging activity. Two plots were subjected to the digging (D1 and D19) and two plots were control (C11 and C18). The sampling took place in five occasions: T0 – before digging; T1 – 14 days; T2 – 45 days; T3 – 75 days; and T4 – 165 days after digging. The results showed no significant difference in the nematode assemblages’ density, diversity and trophic composition between treatments and sampling times, evidencing their high tolerance for naturally stressed environments and to the level of digging they were exposed; Recuperação natural das comunidades de nematodes bentónicos associados aos povoamentos de Zostera noltii após atividade de marisqueio Resumo: As pradarias marinhas são ecossistemas produtivos que suportam elevados níveis de biodiversidade, pelo que estão sujeitos a pressões antropogénicas. Os nematodes são bons indicadores ecológicos pois respondem rapidamente a qualquer perturbação por alterações na densidade, diversidade e estrutura. Este trabalho experimental teve como finalidade o estudo da recuperação natural das comunidades de nematodes associados aos povoamentos de Zostera noltii pela simulação da atividade de marisqueio. Dois plots foram sujeitos a revolvimento (D1 e D19) e dois plots serviram como controlo (C11 e C18) e foram efetuadas amostragens em cinco ocasiões: T0 – antes do revolvimento; T1 – 14 dias; T2 – 45 dias; T3 – 75 dias; e T4 – 165 dias após revolvimento. Os resultados obtidos não mostraram diferenças significativas na diversidade, densidade e composição trófica das comunidades de nematodes entre tratamentos e tempos de amostragem, evidenciando a sua elevada tolerância a ambientes naturalmente dinâmicos e ao nível de revolvimento a que foram expostas.