(2016). Two novel Fusarium species that cause canker disease of prickly ash (Zanthoxylum bungeanum) in northern China form a novel clade with Fusarium torreyae. Mycologia: Vol. 108, No. 4, pp. 668-681.

Oligosaccharide elicitors from pathogens have been shown to play major roles in host plant defense responses involving plant–pathogen chemoperception and interaction. In the present study, chitosan and oligochitosan were prepared from pathogen Fusarium sambucinum, and their effects on infection of Zanthoxylum bungeanum stems were investigated. Results showed that oligochitosan inhibited the infection of the pathogen, and that the oligochitosan fraction with a degree of polymerization (DP) between 5 and 6 showed the optimal effect. Oligochitosan DP5 was purified from fraction DP5-6 and was structurally characterized using electrospray ionization mass spectrometry, Fourier transform infrared spectroscopy, and nuclear magnetic resonance spectroscopy. Oligochitosan DP5 showed significant inhibition against the infection of the pathogenic fungi on host plant stems. An investigation of the mechanism underlying this effect showed that oligochitosan DP5 increased the activities of defensive enzymes and accumulation of phenolics in host Z. bungeanum. These results suggest that oligochitosan from pathogenic fungi can mediate the infection of host plants with a pathogen by acting as an elicitor that triggers the defense system of a plant. This information will be valuable for further exploration of the interactions between the pathogen F. sambucinum and host plant Z. bungeanum.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate the gene expression of target mRNAs involved in plant growth, development, and abiotic stress and pathogen responses. Previous studies have reported miRNAs in Populus that respond to abiotic stresses, such as cold, heat, drought, flooding, high salt and mechanical stress. However, little is known about the regulatory roles of these molecules in the Populus response to the stress of foliar rust fungal infection. Here, we identified the miRNA profiles of Populus after inoculation with Melampsora larici-populina using high-throughput sequencing and bioinformatics analysis. Quantitative real-time PCR (qRT-PCR) was used to validate the expression levels of 10 miRNAs. A total of 90 known miRNAs belonging to 42 families and 378 novel miRNAs were identified from three small-RNA libraries of Populus szechuanica infected with M. larici-populina isolates Sb052 and Th053 and a control. Comparative analysis revealed that the expression of 38 known miRNAs and 92 novel miRNAs in P. szechuanica after infection with different rust fungus isolates showed significant differences, and more miRNAs were suppressed during rust infection. Among the differentially expressed miRNAs, 7 known and 20 novel miRNAs were relevant to the rust fungus infection, and according to KEGG (Kyoto Encyclopaedia of Genes and Genomes) pathway analysis, these miRNAs primarily regulate genes encoding disease-resistance proteins, serine/threonine protein kinases, transcription factors, and related proteins. QRT-PCR analysis indicated that most miRNAs were up-regulated in the Sb052 library and down-regulated in the Th053 library at 48 h post-inoculation (hpi). These results demonstrate that the expression of miRNAs was altered in poplar under stress associated with M. larici-populina infection, and different temporal dynamics were observed in incompatible and compatible libraries. These findings suggest important roles for miRNA regulation in Populus upon infection with foliar rust fungus.

Fusarium zanthoxyli and F. continuum are sister taxa that are the etiological agents of canker disease of prickly ash (Zanthoxylum bungeanum) in northern China. These two pathogens, together with F. torreyae, the causal agent of canker disease of the critically endangered conifer Florida torreya (Torreya taxifolia) from northern Florida and southwestern Georgia, constitute a novel clade, the F. torreyae species complex. To assess their reproductive mode, a polymerase chain reaction (PCR) assay targeting the MAT1-1 and MAT1-2 idiomorphs was designed and validated, using MAT sequences mined from the whole-genome sequence of the three F. torreyae clade pathogens and several closely related fusaria. Results of the MAT idiomorph PCR assay indicated that isolates of the three pathogens were MAT1-1 or MAT1-2. When MAT1-1 and MAT1-2 isolates of each species were crossed on carrot agar, all of the F. zanthoxyli (N = 30) and F. continuum (N = 3) isolates tested were female fertile, yielding mature perithecia with viable ascospores. By comparison, only one pairing of the five different isolates of F. torreyae produced perithecia; however, the majority of the asci in this cross aborted or produced fewer than eight ascospores. Of the three temperatures tested (i.e., 22, 25, and 27 C), the optimal temperature for perithecium production was 22–25 C in F. zanthoxyli and 25 C in F. continuum and F. torreyae. Ascospore progeny from three separate crosses of F. zanthoxyli and F. continuum and one cross of F. torreyae were genotyped to assess whether they were the products of genetic recombination and sexual reproduction. Genotyping of 34–40 progeny from the F. zanthoxyli and F. continuum crosses confirmed that they were the products of sexual reproduction. However, only 36% of the progeny in the F. torreyae cross were recombinant, which was roughly half of the nonparental progeny expected with three markers segregating.

In this report, the effects of two oligochitosans, i.e., oligochitosan A (OCHA) and oligochitosan B (OCHB), on control of dry rot of Zanthoxylum bungeanum (Z. bungeanum) caused by Fusarium sambucinum (F. sambucinum) were evaluated. First, both oligochitosans show desirable ability to decrease the infection of F. sambucinum. Second, the oligochitosans strongly inhibit the radial colony and submerged biomass growth of F. sambucinum. Lastly, these oligochitosans are capable of increasing the activities of phenylalanine ammonia lyase (PAL), polyphenoloxidase (PPO) and peroxidase (POD) significantly, as well as enhancing the content of total phenolics in Z. bungeanum stems. These findings indicate that the protective effects of OCHA and OCHB on Z. bungeanum stems against dry rot may be associated with the direct fungitoxic function against pathogen and the elicitation of biochemical defensive responses in Z. bungeanum stems. The outcome of this report suggests that oligochitosans may serve as a promising natural fungicide to substitute, at least partially, for synthetic fungicides in the disease management of Z. bungeanum.

A wide range of species and hybrids of black and balsam poplars or cottonwoods (Populus L., sections Aigeiros and Tacamahaca) grow naturally, or have been introduced to grow in plantations in China. Many species of Melampsora can cause poplar leaf rust in China, and their distributions and host specificities are not entirely known. This study was prompted by the new susceptibility of a previously resistant cultivar, cv. ‘Zhonghua hongye’ of Populus deltoides (section Aigeiros), as well as by the need to know more about the broader context of poplar leaf rust in China. Rust surveys from 2015 through 2018 in Shaanxi, Sichuan, Gansu, Henan, Shanxi, Qinghai, Beijing, and Inner Mongolia revealed some samples with urediniospores with the echinulation pattern of M. medusae. The morphological characteristics of urediniospores and teliospores from poplar species of the region were further examined with light and scanning electron microscopy. Phylogenetic analysis based on sequences of the rDNA ITS region (ITS₁, 5.8S rRNA gene, and ITS₂) and the nuclear large subunit rDNA (D₁/D₂) was used to further confirm morphology-based identification. Based on combined analyses, five of the fifteen fully characterized samples were identified as Melampsora medusae: one from Shaanxi and four from Sichuan. Two of the five were from Populus deltoides cv. ‘Zhonghua hongye’. Three others were identified on Populus szechuanica, P. simonii, and P. yunnanensis. Additional samples of M. medusae were collected in Shaanxi in 2017 and 2018, and from Henan in 2015 through 2018. Altogether these findings show that this introduced pathogen is widespread and persistent from year to year in China. This is the first report of this North American poplar leaf rust species, Melampsora medusae, in China. It has previously been reported outside North America in Argentina, Europe, Australia, New Zealand, Japan, and Russia.