A Method to Induce Stem Cankers by Inoculating Nonwounded Populus Clones with Septoria musiva Spore Suspensions

Most artificial inoculations of Populus spp. stems with Septoria musiva have required host wounding to induce canker development; in the absence of wounds, frequencies of cankers have been low. Three greenhouse inoculation experiments were conducted to demonstrate the reliability and repeatability of an inoculation method that did not require wounding. In the first, 16 clones of hybrid poplar were inoculated with three isolates of S. musiva to compare responses following wounding and inoculation with mycelium (wound inoculation) with responses following inoculation of nonwounded trees by spraying with a conidial suspension (spray inoculation). Stem disease severity among clones following spray inoculation was correlated with stem disease severity following wound inoculation. A significant clone–isolate interaction was detected with spray inoculation but not wound inoculation. In the other two greenhouse experiments, 29 clones of hybrid poplar and 69 clones of Populus balsamifera were inoculated with a spore suspension mixture of three isolates. In both cases, the experimental error was similar to that obtained in previous experiments, in which trees were wound inoculated, and was adequately small to permit detection of differences in responses among clones. Ultimately, field studies will be needed to determine the best inoculation method for predicting stem responses to this pathogen under field conditions. However, relative to wound inoculation, spray inoculation of nonwounded trees has the advantage of yielding faster results, permitting inoculation with a mixture of isolates, and not circumventing potential mechanisms for resisting penetration. The ability to infect stems without wounding creates opportunities for numerous types of epidemiological and disease control studies that are difficult to conduct with wound inoculation.

Susceptibility of Populus balsamifera to Septoria musiva: A Field Study and Greenhouse Experiment

A greenhouse inoculation experiment and field study were conducted to determine the cause of an outbreak of Septoria musiva cankers on Populus balsamifera in a northern Alberta plantation. Four clones of P. balsamifera, five clones of putatively resistant P. deltoides, and one susceptible hybrid poplar clone, Northwest, were inoculated with seven isolates identified as S. musiva. Four of the isolates were from P. balsamifera in Alberta and the others were from P. deltoides in Quebec. Results indicated that disease severity was similar for Alberta and Quebec isolates (P = 0.243) and that P. balsamifera had the greatest mean disease severity (x-bar = 4.20), P. deltoides had the lowest (x-bar = 2.76), and Northwest was intermediate (x-bar = 3.45). A genetic analysis comparing six polymorphic polymerase chain reaction restriction fragment length polymorphism loci and the mitochondrial small subunit ribosomal DNA sequences of the seven isolates indicated that the Alberta population was made up of at least three distinct genotypes. Canker incidence and age on Northwest and 56 different clones of P. balsamifera in a plantation were recorded. Canker incidence (P = 0.726) and the canker age distributions (P = 0.994) were similar for the two species. In conclusion, contrary to what has been reported in the literature, P. balsamifera appears to be quite susceptible to Septoria canker.

Clone by isolate interaction in the hybrid poplar – Septoria musiva pathosystem

Fourteen clones of hybrid poplar were inoculated with 19 isolates of Septoria musiva Peck under greenhouse conditions to determine the magnitude of the clone, isolate, and clone × isolate interaction effects. Septoria musiva isolates were collected from five geographic areas, two symptoms (canker and leaf spot), and two host types (native species and hybrid poplar). The hybrid poplar clones were classified by parent type ( Populus deltoides Bartr. ex Marsh., Populus laurifolia Lebed. × Populus nigra L., and P. deltoides × (P. laurifolia × P. nigra)). There were no significant differences among geographic areas (p = 0.443), symptoms (p = 0.842), or hosts (p = 0.304) of origin for the 19 isolates nor significant differences among the three parent types (p = 0.089). Clone, isolate, and clone × isolate interaction effects were all significant, accounting for 65%, 15%, and 18%, respectively, of the explained variation. These results indicate that clones rather than parent types should be the focus of resistance screening programs and that the pathosystem should be stable given the relatively small clone × isolate interaction. These results also indicate that a single isolate should be sufficient for preliminary screening of disease resistance in hybrid poplars.

The Histology of Hybrid Poplar Clones Inoculated with Septoria musiva

Septoria musiva causes stem cankers that severely limit production of susceptible hybrid poplars in eastern North America. A field experiment was conducted with resistant clone DN34 and susceptible clone NC11505 in order to (i) identify tissues colonized by the pathogen, (ii) describe tissue responses to S. musiva, and (iii) determine whether tissue responses to S. musiva differed between hybrid poplar clones. Branches of each clone were inoculated by removing the fourth or fifth fully expanded leaf and placing an agar plug colonized by an aggressive isolate of S. musiva over the wound. Seven weeks after inoculation, branches were harvested and prepared for histology. Data from nonwounded control, wounded control, and wounded and inoculated stems were collected and analyzed for effects of clone and treatment. In general, fungal colonization was more extensive in NC11505 and exophylactic and necrophylactic periderms (NPs) of clone DN34 were significantly thicker than those of NC11505, regardless of treatment. The number of NPs produced and the distance from the epidermis to the outermost layer of phellem were significantly affected by the pathogen. Inoculated stems of clone DN34 developed a single NP that formed closer to the wound surface than in wounded controls. In contrast, inoculated stems of NC11505 developed successive NPs and the first NP formed further from the wound surface than in wounded controls. These two host responses to inoculation, as well as measures of exophylactic and necrophylactic periderm thickness, may be useful as markers for the selection of poplar resistant to damage by S. musiva.

Evaluating the interaction between genotype and water stress in the hybrid poplar – Septoria musiva pathosystemThis note is one of a selection of papers published in the Special Issue on Poplar Research in Canada.

Septoria musiva , the causal agent of Septoria canker, has caused widespread damage to plantations of hybrid poplar across North America. A greenhouse experiment was conducted to evaluate the size of interactions between the genotype factors of (i) clone, (ii) isolate, and (iii) the clone–isolate interaction and the environmental factor of water stress. Four clones of hybrid poplar were inoculated with four isolates of S. musiva and subsequently exposed to two levels of water stress (stressed and unstressed). Tree height, root collar diameter (RCD), and disease severity were measured 56 d after inoculation. Water stress reduced height and RCD growth by approximately 30% but did not affect disease severity (P = 0.258). Of the explained variability, 97.5% was attributable to genotype effects (89% from clone, 0% from isolate, and 8.5% from clone–isolate interaction). The remaining 2.5% of the variation was accounted for by the genotype – water stress interaction. These results suggest that clonal effects will be more important determinants of disease severity under greenhouse conditions than genotype – water stress interactions.