Martina Cerri

Demographic and macro-morphological evidence for common reed dieback in central Italy

Background: Phragmites australis dieback syndrome, recently reported for the Mediterranean basin, features several symptomatic traits among which the clumping habit seems to be one of the most diagnostic. Aims: We evaluated the effect of water depth on the intensity of the clumping habit and evaluated the diagnostic role of other traits. Methods: We investigated patterns of macro-morphological (culm height and diameter, flowering head and dead apical bud rates) and demographic (density) traits of P. australis in function of submersion (PF-permanent vs. NF-temporary) at five Italian wetland sites. We related the occurrence of clumping and its frequency with water depth. Results: There were clear trends, modulated by site-specific effects, for most of the considered traits in function of the duration of submersion. The clumping rate was close to zero in NF-stands, reached high values in PF-stands and was positively correlated with water depth. Conclusions: We have shown that permanent submersion with deep water levels plays a crucial role in the occurrence of reed dieback. As all other considered traits, with the exception of the occurrence of dead apical buds, well correlate with the clumping habit, we propose using clumping as a key indicator for detecting potential reed dieback.

AFLP Approach Reveals Variability in Phragmites australis: Implications for its Die-back and Evidence for Genotoxic Effects

Phragmites australis is a subcosmopolitan species typical of wetlands being studied in Europe for its disappearance from natural stands, a phenomenon called reed die-back syndrome (RDBS). Although it is conjectured that low genetic variability contributes to RDBS, this aspect remains neglected to this day. Using a molecular fingerprinting approach and a sequence analysis of the trnT-trnL/rbcL-psaI regions of cpDNA, this study aimed to compare the genetic structure of stable vs. RDBS-affected P. australis stands from five wetlands of central Italy. Beforehand, in order to characterize the health condition of reed populations, the occurrence of the main macromorphological descriptors for RDBS was considered on 40 reed stands. Soil samples were also collected to examine the total content of heavy metals. The current study analyzed cpDNA in 19 samples and AFLP profiles in 381 samples to investigate the genetic structure of Phragmites populations. Based on the multinomial-Dirichlet model, an analysis of candidate loci under selective pressure was also performed. The relationships among AFLP data, RDBS descriptors and chemicals were evaluated with the use of Linear Mixed Models. The analysis of the cpDNA shows the occurrence of the haplotypes M (the most widespread), and K here recorded for the first time in Italy. Three new haplotypes were also described. The DNA fingerprinting analysis has produced a total of 322 loci (98% polymorphic) and shows the medium-to-high amount of genetic diversity. The significant genetic differentiation among wetlands (Fst = 0.337) suggests either low gene flow or small effective population size. Moreover, the low amount of outlier loci (only 5; l.5% of the total), seems to indicate the scarce occurrence of selective pressure upon the reed’s genome. Genetic diversity increased in relationship to the decrease in diameter and of flowering buds of the reed, two of the trends associated with the die-back. The current study rejects the hypothesis that genetic diversity massively contributed to RDBS. Moreover, significant relationships between genetic diversity and the total concentration of some heavy metals (Cr, Cu, and Zn) were highlighted, indicating possible genotoxic effects on P. australis. The current study represents a fact-finding background useful for the conservation of common reed.

Claviceps arundinis identification and its role in the die-back syndrome of Phragmites australis populations in central Italy

Abstract Common reed Phragmites australis (Cav.) Trin. ex Steud. is one of the most widely distributed angiosperms with important ecological functions. In recent decades, it has been affected by a severe decline known as reed die-back syndrome (RDBS), the causal factors of which are still under investigation. Among the biotic factors that influence the dynamic of the reed population, the role of microorganisms is still poorly understood. During surveys carried out on P. australis populations in Central Italy, Claviceps-like sclerotia were detected: is Claviceps infection related to P. australis sexual reproduction and seed production? Could Claviceps infection be involved in the RDBS? These are the questions that we address. We characterized the sclerotia at the morphological, molecular, and chemical level and we demonstrated that they belong to Claviceps arundinis Pažoutová & M. Kolařík. To our knowledge, this is the first report of C. arundinis on P. australis in Italy. Furthermore, the association of C. arundinis with RDBS was evaluated considering a set of macromorphological traits generally related to RDBS, such as P. australis clumping habit, culm height and diameter. No correlation was seen between the occurrence of C. arundinis and the declining status of reed populations.

Oomycete Communities Associated with Reed Die-Back Syndrome

Phragmites australis (Cav.) Trin. ex Steud. die-back is a widely-studied phenomenon that was first discovered in northern Europe and that, until recently, was almost unknown in the Mediterranean basin. It has been described as a complex syndrome affecting reed populations leading to their retreat and decline, with significant impacts on valuable ecosystem services. Among the factors that cause the decline, soil-living microorganisms can be crucial. The aims of this study were to analyze the diversity of oomycetes communities associated with reed stands, and to understand whether they could play a key role in the decline. Variations in the structure of oomycetes communities were studied by metabarcoding of the internal transcribed spacer (ITS) 1 region of ribosomal DNA, from the sediments of five Italian freshwater ecosystems. They were chosen to cover a large variability in terms of surface area, water depth, microclimate, and presence of documented reed retreat. From 96 samples collected from reed roots, rhizosphere, and bulk soil, we assembled 207661 ITS1 reads into 523 OTUs. We demonstrated that oomycete communities were structured by several factors, among which the most important was die-back occurrence. Our study also indicates that Pythiogeton spp. could be potentially involved in the development of die-back. The role of heavy metals in the soil was also explored, and cadmium concentration was shown to affect oomycetes distribution. This study represents a significant step forward for the characterization of microbial communities associated with reed die-back syndrome and helps to gain knowledge of the complexity of these important wet ecosystems.

Pectobacterium aroidearum and Pectobacterium carotovorum subsp. carotovorum as causal agents of potato soft rot in Lebanon

To identify bacteria causing severe soft rot on potato in Lebanon, pectinolytic bacteria were isolated from diseased samples collected in the Bekaa valley and the Akkar region, during 2009–2010. Six isolates that induced a hypersensitive reaction in tobacco plants and caused a marked soft rot when inoculated in potato tubers or stem, DAPP-PG 752 to DAPP-PG 757, were further characterized. Analysis of 16S–23S intergenic transcribed spacer-regions, digested with RsaI and CfoI, demonstrated that isolates DAPP-PG 753 to DAPP-PG 757 showed electrophoretic profiles similar to that of LMG 2408, a strain misclassified as Pectobacterium carotovorum subsp. carotovorum, and different from that of DAPP-PG 752. Phylogenetic analysis using partial malate dehydrogenase gene sequences, clustered DAPP-PG 752 in a clade that contained the type strain of P. carotovorum subsp. carotovorum, while DAPP-PG 753 to DAPP-PG 757, together with LMG 2408, fell in a clade including strains of the recently described species Pectobacterium aroidearum. Multilocus sequence analysis of concatenated partial sequences of the atpD, gyrB, infB and rpoB genes confirmed the results obtained with the mdh gene sequences and identified DAPP-PG 752 as P. carotovorum, and LMG 2408 and the five related Lebanese isolates as P. aroidearum. The Lebanese P. aroidearum isolates were phenotypically similar to known P. aroidearum strains, based on API 50 CHE data. They showed identical BOX- and ERIC-PCR profiles that resembled profiles of known P. aroidearum strains. This study represents the first official report of P. aroidearum as causal agent of soft rot on potato plants and the first occurrence of P. carotovorum subsp. carotovorum on potato plants in Lebanon.

Influence of die-back syndrome on reproductive strategies within Phragmites australis populations

Abstract Sexual plant reproduction is a strategy that allows plant populations to increase genetic variability, and consequently to be more efficient in adapting to new environments and to overcome stress conditions. Here, we focus on the reproductive mode of Phragmites australis, an important sub-cosmopolite species that can spread both by clonal propagation and sexual reproduction. In Europe, P. australis is affected by severe decline (known as Reed Die-Back Syndrome or RDBS); this study aims to understand if in RDBS condition the reproductive strategy of P. australis was altered, based on the hypothesis that if stress occurs, plants are likely to use their energy to increase genetic variability to overcome the stress. Inflorescences in five Italian wetlands were sampled from healthy and RDBS-affected stands of P. australis. Seed production, seed viability and seed germination were evaluated, with germination monitored under two different water conditions: moist and submerged. Stress induced by RDBS appears to influence the reproductive strategy of P. australis. In RDBS-affected stands, seed production was significantly higher than healthy stands for four of five sites. Seed viability and germination were also higher in RDBS-affected stands, but these differences were lower and site dependent.

Spatial landscape patterns and trends of declining reed-beds in peninsular Italy

Abstract Patterns and trends of declining reed-beds in four freshwater ecosystems in central Italy are investigated through an aerial orthoimagery-based diachronic analysis over a period of 24 years. Extent variations and retreat from the waterfront are detected in all sites, compensated only in few cases by backwards enlargements. These shape and size modifications suggest that reed die-back is associated with retreat and with a notable fragmentation process. Long-term analysis performed in one of the sites, compared to water levels, shows a drastic extent decrease co-occurring with an artificial rise of the water level, confirming the regulation of freshwater bodies as a driver of biodiversity loss. Results also suggest that reed-dominated ecosystems may reach a threshold of tolerance towards stressing conditions beyond which they cannot regenerate. The carried out study testifies for a valuable role of landscape metrics in analysing the spatial processes related to declining reed-beds. Spatial metrics prove to be a suitable tool in monitoring processes, working as early warning signals of ongoing decline and helping to model future changes.