Biocontrol of Cytospora canker of poplar in north-east China with Trichoderma longibrachiatum

Trichoderma isolate T05, collected and isolated from a poplar (Populus simonii x Populus nigra) stand in the province of Heilongjiang, China, was tested for its efficacy in controlling Cytospora chrysosperma in vitro and Cytospora canker of poplar in the field. Trichoderma isolate T05 was identified as Trichoderma longibrachiatum in sect. Longibrachiatum based on both morphological characteristics and phylogenetic analysis of the nuclear ribosomal internal transcribed spacer (ITS1 and ITS2) oligonucleotide sequences and the short intron of tef1. During dual culturing, the inhibitory effect of T. longibrachiatum T05 on C. chrysosperma increased over time, and inhibition up to 81% was observed 120 h after inoculation. Hyphae of T. longibrachiatum T05 grew alongside, coiled and sporulated around the pathogen hyphae, which were generally observed to be wrinkled, collapsed, disrupted and degraded. Culture filtrates of T. longibrachiatum T05 that were sterilized by either bacterial filtration or autoclaving suppressed the growth of the pathogen colony. Filtrate sterilized by filtration showed greater inhibitory activity than the autoclaved filtrate, which still had some inhibitory activity, demonstrating that some inhibitory substances were heat unstable and some heat stable. Cytospora chrysosperma C01 colony growth was also reduced by volatile antibiotics produced by T. longibrachiatum T05, with inhibition of 54% observed after 96 h. In a field trial, Cytospora canker on poplar trunks was controlled with efficacy similar to that of routine chemical control by applying a T. longibrachiatum preparation.