One isolate each of Trichoderma viride, Epicoccum nigrum, Fusarium tricinctum, Alternaria alternata, Sclerotinia sclerotiorum and Cytospora (teleomorph: Valsa sp.) present in epigeous declining oak tissues was evaluated for its ability to control Diplodia corticola (isolate 79). This fungus is the causal agent of cankers, vascular necrosis and dieback on various oak species. Among the isolates tested, T. viride and F. tricinctum showed maximum in vitro inhibition of mycelial growth of D. corticola (isolate 79). Species were also evaluated for their ability to reduce mortality caused by D. corticola (isolate 79) of Quercus cerris and Q. pubescens seedlings under controlled conditions. Two series of inoculations were carried out through wounds in the stem; in the first, the distance between the point of inoculation of the antagonist and the pathogen was 6 cm, whereas in the second series the distance was shortened to 3 cm. In seedlings of Q. cerris and Q. pubescens at a distance of 3 cm, inoculation with F. tricinctum and A. alternata significantly reduced mortality caused by D. corticola (isolate 79). Inoculation of T. viride through artificial cuticular wounds in the stem of seedlings prevented the proliferation of D. corticola (isolate 79) only on seedlings of Q. cerris. All Q. pubescens seedlings treated with T. viride manifested pathological symptoms subsequent to proliferation of D. corticola (isolate 79). These observations indicate that the interactions between endophytes in planta and D. corticola (isolate 79) are complex and merit further study.
Fuscoporia torulosa is the causal agent of white alveolar wood decay on several species including a large number of forest trees. Early detection of the fungus is essential to identify diseased trees before spread occurs to healthy plants. However, current detection methods based on isolation from infected tissues on semi-selective media are laborious, time consuming and require expertise in identifying the pathogen after isolation. In the present study, a rapid and reliable Scorpion-PCR based molecular method to identify and detect F. torulosa in planta was developed in a highly polymorphic portion of the internal transcribed spacer (ITS) regions. Specificity of primers and probe was assessed by means of both BLAST analyses and by using genomic DNA from 131 F. torulosa isolates and 43 other fungi and oomycetes from different hosts and geographic areas. In Scorpion-PCR the limit of detection was 1 pg of total DNA and a high correlation ( r 2 = 0·996)was achieved between target DNA quantity and cycle threshold (Ct). Real-time PCR combined with effective procedures for DNA extraction enabled the detection of F. torulosa from naturally infected tissues of oaks with and without fruit bodies in approximately 6 h. Comparative testing showed that detection of F. torulosa in wood samples is more sensitive and reliable with real-time PCR than with conventional isolation.
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