Transcriptomic analysis of molecular responses in Malus domestica 'M26' roots affected by apple replant disease.

KEY MESSAGE: Gene expression studies in roots of apple replant disease affected plants suggested defense reactions towards biotic stress to occur which did not lead to adequate responses to the biotic stressors. Apple replant disease (ARD) leads to growth inhibition and fruit yield reduction in replanted populations and results in economic losses for tree nurseries and fruit producers. The etiology is not well understood on a molecular level and causal agents show a great diversity indicating that no definitive cause, which applies to the majority of cases, has been found out yet. Hence, it is pivotal to gain a better understanding of the molecular and physiological reactions of the plant when affected by ARD and later to overcome the disease, for example by developing tolerant rootstocks. For the first time, gene expression was investigated in roots of ARD affected plants employing massive analysis of cDNA ends (MACE) and RT-qPCR. In reaction to ARD, genes in secondary metabolite production as well as plant defense, regulatory and signaling genes were upregulated whereas for several genes involved in primary metabolism lower expression was detected. For internal verification of MACE data, candidate genes were tested via RT-qPCR and a strong positive correlation between both datasets was observed. Comparison of apple 'M26' roots cultivated in ARD soil or γ-irradiated ARD soil suggests that typical defense reactions towards biotic stress take place in ARD affected plants but they did not allow responding to the biotic stressors attack adequately, leading to the observed growth depressions in ARD variants.

Impaired defense reactions in apple replant disease-affected roots of Malus domestica 'M26'.

A soil- and site-dependent complex of diverse microbial populations causes apple replant disease (ARD), which leads to economic losses for tree nurseries and apple producers due to reduced plant growth and diminished fruit yields. Soil fumigation has been widely used to mitigate ARD, but the application of these chemicals is restricted in the European Union. Hence, other counteractions have to be developed. Genomics-based breeding may be used to select ARD-tolerant genotypes; however, molecular responses of ARD are not well understood. Recent studies revealed that biotic stress-associated genes involved in typical defense reactions are activated but do not result in an adequate response to ARD. The objective of this study was to analyze selected responsive genes in a time-course experiment to test for expression kinetics. Cultivating the ARD-susceptible apple rootstock 'M26' on ARD-affected soil resulted in significantly reduced growth as early as 7 days after planting. Genes involved in phytoalexin biosynthesis were upregulated in ARD samples as early as 3 days after planting and reached up to 26-fold changes at Day 10, which resulted in high amounts of 3-hydroxy-5-methoxybiphenyl, aucuparin, noraucuparin, 2-hydroxy-4-methoxydibenzofuran, 2'-hydroxyaucuparin and noreriobofuran. For the first time, these phytoalexins were detected, identified and quantified in apple roots. The lack of a sufficient defense response may be due to impaired sequestration and/or exudation of the potentially cytotoxic phytoalexins and perturbed formation of reactive oxygen species, leading to root damage in ARD soils. The findings provide a basis for comparative studies of the defense processes in more ARD-tolerant rootstocks.

Associação entre otites bacterianas e infecção pelo circovírus suíno tipo 2 (PCV2) em suínos / Association between bacterial otitis and porcine circovirus 2 (PCV2) infection in pigs

Neste trabalho, foi estudada a ocorrência e a bacteriologia das otites purulentas em suínos apresentando três condições clínico-patológicas distintas: síndrome multissistêmica do definhamento dos suínos (SMDS), crescimento retardado (porém sem apresentação de sinais clínicos da SMDS) e crescimento normal. No total, foram examinados 385 animais com idade de 60-130 dias, sendo a confirmação do diagnóstico da SMDS obtida pelos sinais clínicos e achados patológicos e pela detecção de antígenos virais por imunoistoquímica. De 242 animais com a SMDS, 57 (23,5 por cento) apresentaram lesões purulentas no ouvido médio. Entre os 119 animais de baixo desenvolvimento, apenas 1 (0,7 por cento) apresentou a lesão. Não foram detectadas lesões macroscópicas no ouvido médio dos 24 animais com crescimento normal. Os agentes isolados com maior freqüência foram Arcanobacterium pyogenes (43 por cento), Streptococcus α-hemolíticos (37,2 por cento) e Pasteurella multocida (27,9 por cento) dos 86 ouvidos submetidos à bacteriologia. A alta ocorrência de lesões purulentas no ouvido médio de animais com a SMDS sugere que a infecção pelo PCV2 pode tornar o suíno mais suscetível às otites bacterianas. A ocorrência reduzida dessas lesões em suínos de baixo desenvolvimento, sem a manifestação clínica da SMDS, sugere que a otite média não representa uma causa importante de mau desempenho em leitões nas fases de crescimento e terminação. O envolvimento de A. pyogenes, Streptococcus α-hemolíticos e P. multocida na maioria das lesões apontam a importância desses organismos como agentes causais de otite média em suínos.(AU)

The occurrence and bacteriology of purulent otitis in groups of pigs affected by three different clinical conditions are described: postweaning multisystemic wasting syndrome (PMWS), depressed growth (attrition) and normal growth (control). A total of 385 animals, 60-130 days of age, were examined. Diagnosis of PMWS was confirmed by the presence of clinical signs and pathological findings compatible with the disease, and through detection of viral antigens in tissues by immunohistochemistry. From 242 pigs with PMWS, 57 (23.5 percent) showed purulent lesions in the middle ear. Among 119 pigs with attrition, only 1 (0.7 percent) presented the lesion. In 24 control pigs, middle ear lesions were not detected. The most frequently isolated agents from the lesions were Arcanobacterium pyogenes (43 percent), á-hemolytic Streptococci (37.2 percent) and Pasteurella multocida (27.9 percent) out of 86 materials. The frequent occurrence of purulent lesions in the middle ear of PMWS affected pigs suggests that PCV2 infection may increase the susceptibility to bacterial otitis. The low occurrence of this disease in piglets with attrition suggests that otitis does not represent a significant cause for depressed growth in pigs from growing and finishing age. The isolation of A. pyogenes, á-hemolytic Streptococci and P. multocida from most of the lesions emphasizes the importance of these organisms as causal agents of otitis media in swine.(AU)