Abstract. Various features of the biology of the rust fungi and of the epidemiology of the plant diseases they cause illustrate the important role of rainfall in their life history. Based on this insight we have characterized the ice nucleation activity (INA) of the aerially disseminated spores (urediospores) of this group of fungi. Urediospores of this obligate plant parasite were collected from natural infections of 7 species of weeds in France, from coffee in Brazil and from field and greenhouse-grown wheat in France, the USA, Turkey and Syria. Immersion freezing was used to determine freezing onset temperatures and the abundance of ice nuclei in suspensions of washed spores. Microbiological analyses of spores from France, the USA and Brazil, and subsequent tests of the ice nucleation activity of the bacteria associated with spores were deployed to quantify the contribution of bacteria to the ice nucleation activity of the spores. All samples of spores were ice nucleation active, having freezing onset temperatures as high as −4 °C. Spores in most of the samples carried cells of ice nucleation-active strains of the bacterium Pseudomonas syringae (at rates of less than 1 bacterial cell per 100 urediospores), but bacterial INA accounted for only a small fraction of the INA observed in spore suspensions. Changes in the INA of spore suspensions after treatment with lysozyme suggest that the INA of urediospores involves a polysaccharide. Based on data from the literature, we have estimated the concentrations of urediospores in air at cloud height and in rainfall. These quantities are very similar to those reported for other biological ice nucleators in these same substrates. However, at cloud level convective activity leads to widely varying concentrations of particles of surface origin, so that mean concentrations can underestimate their possible effects on clouds. We propose that spatial and temporal concentrations of biological ice nucleators active at temperatures > −10 °C and the specific conditions under which they can influence cloud glaciation need to be further evaluated so as to understand how evolutionary processes could have positively selected for INA.

Abstract. For decades, drop-freezing instruments have contributed to a better understanding of biological ice nucleation and its likely implications for cloud and precipitation development. Yet, current instruments have limitations. Drops analysed on a cold stage are subject to evaporation and potential contamination. The use of closed tubes provides a partial solution to these problems, but freezing events are still difficult to be clearly detected. Here, we present a new apparatus where freezing in closed tubes is detected automatically by a change in light transmission upon ice development, caused by the formation of air bubbles and crystal facets that scatter light. Risks of contamination and introduction of biases linked to detecting the freezing temperature of a sample are then minimized. To illustrate the performance of the new apparatus we show initial results of two assays with snow samples. In one, we repeatedly analysed the sample (208 tubes) over the course of a month with storage at +4 • C, during which evidence for biological ice nucleation activity emerged through an increase in the number of ice nucleators active around −4 • C. In the second assay, we indicate the possibility of increasingly isolating a single ice nucleator from a precipitation sample, potentially determining the nature of a particle responsible for a nucleation activity measured directly in the sample. These two seminal approaches highlight the relevance of this handy apparatus for providing new points of view in biological ice nucleation research.

Pseudomonas syringae is the most frequently emerging group of plant pathogenic bacteria. Because this bacterium is ubiquitous as an epiphyte and on various substrates in non-agricultural settings, there are many questions about how to assess the risk for plant disease posed by strains in the environment. Although P. syringae is considered to have discrete host ranges in defined pathovars, there have been few reports of comprehensive comparisons of host range potential. Here we present results of host range tests for 134 strains, representing eight phylogroups, from epidemics and environmental reservoirs on 15 to 22 plant species per test conducted in four separate tests to determine the patterns and extent of host range. We sought to identify trends that are indicative of distinct pathotypes and to assess if strains in the P. syringae complex are indeed restricted in their host range. We show that for each test, strains display a diversity of host ranges from very restricted to very broad regardless of the gamut of phylogroups used in the test. Overall, strains form an overlapping continuum of host range potential with equal representation of narrow, moderate and broad host ranges. Groups of distinct pathotypes, including strains with currently the same pathovar name, could not be identified. The absence of groupings was validated with statistical tests for pattern recognition. The extent of host range was positively correlated with the capacity of strains to swarm on semi-solid agar medium and with the abundance of genes in biosynthetic clusters and was inversely correlated with the abundance of genes for proteins with transmembrane domains in their genomes. Our results are consistent with the current paradigm that disease symptoms are the result of multiple molecular interactions between P. syringae and its plant host that are modulated by abiotic and biotic conditions. This leads us to propose that pathovar denominations do not correspond to the underlying biology of P. syringae. A new concept of pathogenicity that accounts for the continuum of pathogenic potential in P. syringae would open new perspectives to understand the evolution of pathogenicity in this bacterium and new insights to anticipate disease and to manage plant health.

Outdoor field measurements of bioaerosols are performed within a wide range of basic and applied scientific disciplines, each with its own goals, assumptions, and terminology. This article contains brief reviews of outdoor field bioaerosol research from these diverse interests, with emphasis on perspectives from the atmospheric sciences. The focus is on a high-level discussion of pressing scientific questions, grand challenges, and needs for cross-disciplinary collaboration. The research topics, in which bioaerosol field measurement is important, include (i) atmospheric physics, clouds, climate, and hydrological cycle; (ii) atmospheric chemistry; (iii) airborne allergencontaining particles; (iv) airborne human pathogens and national security; (v) airborne livestock and crop pathogens; and (vi) biogeography and biodiversity. We concisely review bioaerosol impacts and discuss properties that distinguish bioaerosols from abiological aerosols. We give extra focus to regions of specific interest, i.e., forests, polar regions, marine and coastal environments, deserts, urban and rural areas, and summarize key considerations related to bioaerosol measurements, such as of fluxes, of long-range transport, and of sampling from both stationary and vessel-driven platforms. Keeping in mind a series of key scientific questions posed within the diverse communities, we suggest that pressing scientific questions include the following: (i) emission sources and flux estimates; (ii) spatial distribution; (iii) changes in distribution; (iv) atmospheric aging; (v) metabolic activity; (vi) urbanization of allergies; (vii) transport of human pathogens; and (viii) climate-relevant properties.

Abstract. Some biological particles and macromolecules are particularly efficient ice nuclei (IN), triggering ice formation at temperatures close to 0 ∘C. The impact of biological particles on cloud glaciation and the formation of precipitation is still poorly understood and constitutes a large gap in the scientific understanding of the interactions and coevolution of life and climate. Ice nucleation activity in fungi was first discovered in the cosmopolitan genus Fusarium, which is widespread in soil and plants, has been found in atmospheric aerosol and cloud water samples, and can be regarded as the best studied ice-nucleation-active (IN-active) fungus. The frequency and distribution of ice nucleation activity within Fusarium, however, remains elusive. Here, we tested more than 100 strains from 65 different Fusarium species for ice nucleation activity. In total, ∼11 % of all tested species included IN-active strains, and ∼16 % of all tested strains showed ice nucleation activity above −12 ∘C. Besides Fusarium species with known ice nucleation activity, F. armeniacum, F. begoniae, F. concentricum, and F. langsethiae were newly identified as IN-active. The cumulative number of IN per gram of mycelium for all tested Fusarium species was comparable to other biological IN like Sarocladium implicatum, Mortierella alpina, and Snomax®. Filtration experiments indicate that cell-free ice-nucleating macromolecules (INMs) from Fusarium are smaller than 100 kDa and that molecular aggregates can be formed in solution. Long-term storage and freeze–thaw cycle experiments revealed that the fungal IN in aqueous solution remain active over several months and in the course of repeated freezing and thawing. Exposure to ozone and nitrogen dioxide at atmospherically relevant concentration levels also did not affect the ice nucleation activity. Heat treatments at 40 to 98 ∘C, however, strongly reduced the observed IN concentrations, confirming earlier hypotheses that the INM in Fusarium largely consists of a proteinaceous compound. The frequency and the wide distribution of ice nucleation activity within the genus Fusarium, combined with the stability of the IN under atmospherically relevant conditions, suggest a larger implication of fungal IN on Earth’s water cycle and climate than previously assumed.