Blueberries (Vacciniumspp.) have been widely cultivated in China because of their nutritional benefits and economic value. Blueberry stem blight has become one of the most severe diseases influencing blueberry productivity and quality in China. In this study, eight fungal isolates were obtained from twenty stem blight lesions of blueberry collected in Nanping, Fujian province, China. Asexual stage was observed after inducing sporulation, the morphology of which agrees withMacrophominain the black, smooth, hard sclerotia and ellipsoid to obovoid, smooth hyaline conidia with apical sheath. Furthermore, DNA sequences of concatenated ITS,tef1-α,TUB, andACTloci indicated that these isolates belong to a novel fungal species. The distinguishing morphological characteristics, such as the wider conidia and larger conidiomata pycnidial, also support its new status. Thus a novel fungus,Macrophominavaccinii, was described in this study. Pathogenicity tests indicated thatM.vacciniicould cause stem blight of blueberry.
Blueberries (Vaccinium spp.) are largely cultivated in China because of their nutritional benefits and economic value. Blueberry stem blight caused by members of the Botryosphaeriaceae has become one of the most severe diseases affecting blueberry cultivation in China. In this study, we examined the causal agent of blueberry stem blight at commercial greenhouse farms in the suburban area of Beijing, China. In total, 37 isolates of Botryosphaeriaceae were obtained from 100 stem blight samples of blueberry. Twelve of 37 strains were morphologically consistent with the genus Lasiodiplodia, showing ellipsoid to ovoid, one-celled, hyaline conidia that sometimes turned brown, with median septa and longitudinal striations when mature. These 12 strains were identified as belonging to a novel fungal species, Lasiodiplodia vaccinii, based on phylogenetic analysis of the concatenated internal transcribed spacer, RNA polymerase II gene, β-tubulin gene, and translation elongation factor-1α gene sequences as well as morphological characteristics. Pathogenicity tests indicated that L. vaccinii can cause twig blight on blueberry seedlings in the greenhouse. Mycelial growth of L. vaccinii occurred at pH values ranging from 3.0 to 10.0, with an optimum at 6.2, and at temperatures from 15 to 40°C, with an optimum at 30.3°C. Of the seven carbon sources tested, sucrose, fructose, and glucose were all highly efficient in supporting the mycelial growth of L. vaccinii, and xylose was the least effective. Of six nitrogen sources tested, yeast extract and tryptone best promoted mycelial growth of L. vaccinii. The ability of L. vaccinii to grow at high temperatures may help to explain its occurrence in Beijing greenhouses in this study.
Marssonina leaf spot disease of poplar (MLDP), caused by the hemibiotrophic pathogen Marssonina brunnea, frequently results in damage to many poplar species. In nature, two formae speciales of M. brunnea exist that are susceptible to different poplar subgenera. Marssonina brunnea f. sp. monogermtubi infects poplar hosts from Populus sect. Aigeiros (Aig), while M. brunnea f. sp. multigermtubi always infects poplar hosts from Populus sect. Leuce Duby (Leu). Based on the fungal penetration structures, a comprehensive transcriptomic approach was used to investigate the gene expression patterns of these two poplar subgenera at three crucial infection stages. MLDP significantly altered the expression patterns of many genes involved in mitogen activated protein kinase (MAPKs) and calcium signaling, transcription factors, primary and secondary metabolism, and other processes in both poplar subgenera. However, major differences in gene expression were also observed between the two poplar subgenera. Aig was most responsive at the initial infection stage, while Leu largely interacted with M. brunnea at the necrotrophic phase. Furthermore, the differentially expressed genes (DEGs) involved in pathways related to biotic stress also differed substantially between the two poplar subgenera. Further analysis indicated that the genes involved in cell wall metabolism and phenylpropanoid metabolism were differentially expressed in the progression of the disease. By examining the expression patterns of genes related to the defense against disease, we found that several genes annotated with causing hypersensitive cell death were upregulated at the necrotrophic phase of MLDP, inferring that plant immune response potentially happened at this infection stage. The present research elucidated the potential molecular differences between the two susceptible interaction systems in MLDP and provided novel insight into the temporal regulation of genes during the susceptible response. To the best of our knowledge, this study also constitutes the first to reveal the molecular mechanisms of poplar in response to the transition of hemibiotrophic fungal pathogens from the biotrophic phase to the necrotrophic phase.
Poplar, which is a dominant species in plant communities distributed in the northern hemisphere, is commonly used as a model plant in forestry studies. Poplar production can be inhibited by infections caused by bacteria, including Lonsdalea quercina subsp. populi, which is a gram-negative bacterium responsible for bark canker disease. However, the molecular basis of the pathogenesis remains uncharacterized. In this study, we annotated the two-component signal transduction systems (TCSs) encoded by the L. quercina subsp. populi N-5-1 genome and identified 18 putative histidine kinases and 24 response regulators. A large-scale mutational analysis revealed that 19 TCS genes regulated bacterial virulence against poplar trees. Additionally, the deletion of kdpE or overexpression of kdpD resulted in almost complete loss of bacterial virulence. We observed that kdpE and kdpD formed a bi-cistronic operon. KdpD exhibited autokinase activity and could bind to KdpE (Kd = 5.73 ± 0.64 μM). Furthermore, KdpE is an OmpR family response regulator. A chromatin immunoprecipitation sequencing analysis revealed that KdpE binds to an imperfect palindromic sequence within the promoters of 44 genes, including stress response genes Lqp0434, Lqp3037, and Lqp3270. A comprehensive analysis of TCS functions may help to characterize the regulation of poplar bark canker disease.