Bionomics and host plants of the invasive Cydia interscindana (Möschler, 1866) (Lepidoptera, Tortricidae), an emerging pest in the Carpathian Lowlands

Cydia interscindana (Möschler, 1866) has spread through several European countries in the past few years, becoming an invasive pest of ornamental trees. It was collected in Hungary for the first time in a pheromone trap set for Cydia pomonella (Linnaeus, 1758) in 2014. Here we discuss its recent distribution in Hungary based on intensive sampling between 2018 and 2020, which showed the dispersal of the pest by humans. Two formerly unknown host plants are also recorded. The damage caused by the larvae, the external morphology of the adult male, larva, pupa (described for the first time) and pupal exuviae are presented. We also analyse DNA barcodes, identifying this pest for the first time via DNA sequencing of immature stages. Introduction Cydia interscindana is native in the Mediterranean region, where it was described by Möschler in 1866 from Andalusia. It is distributed in Mediterranean countries including Portugal (Corley 2004), Spain (Férriz et al. 2006), France (Lévêque et al. 2017) and Italy (Minelli 1995). Later the species was recorded in the British Isles (Knill-Jones 2020), Belgium (De Prins 2016), Switzerland (Swisslepteam 2010), Slovakia (Pastorális et al. 2018) and Russia (Caucasus; Schurov et al. 2017). In Hungary, Cydia interscindana adults were caught by a sticky delta pheromone trap (CSALOMON RAG type) for Cydia pomonella (Linnaeus 1758) in 2014 during a study on swarming dynamics of the latter pest in Budapest. This provided the first record of the species in the Carpathian basin (Szabóky 2014; Takács and Szabóky 2015). In the Mediterranean region larvae feed on Juniperus oxycedrus (L.) (Miller 1990). In Belgium the larva was recorded on Juniperus spp. (Meert et al. 2019). J. oxycedrus is not native in Hungary, but Cupressus × leylandii A.B. Jacks. & Dallim 1926, Platycladus orientalis (L.) Franco 1949 and Chamaecyparis lawsoniana (A. Murray bis) Parl. 1864 are popular evergreens used as ornamental trees both in parks and gardens. In Hungary several pests of these plants have been recorded, all probably introduced with imported plants; in the literature, 11 Lepidoptera, nine Coleoptera and six Hemiptera species have been mentioned already (Csóka and Kovács 1999; Maráczi 2013; Bozsik et al. 2016; Schurov et al. 2017). However, until the end of the 2000s, only Scolytidae (Coleoptera) species caused serious damage (Bozsik and Szőcs 2017). In 2012, an outbreak of the formerly detected (Muskovits 2001) Lamprodila festiva (Linnaeus 1767) (Buprestidae) took place in Budapest (Németh 2012) causing serious damage on Platycladus orientalis and several ornamental gymnosperm species. This outbreak was certainly caused by introduced specimens, that had arrived with trees from the Mediterranean region where this beetle is a well-known pest (Merkl 2016), whose abundance in Hungary increases due to climatic change (Csóka et al. 2018). Based on the available data, in Hungary this beetle pest has also been blamed for all the damage caused on Cupressus, Platycladus and Chamaecyparis trees and management has been carried out only against them. In 2018, a larva of L. festiva, an unidentified caterpillar and a freshly emerged specimen of Cydia interscindana were collected simultaneously from a Leyland cypress in Székesfehérvár (Central Hungary). In that year, similar Lepidoptera larvae were found in three neighbouring villages: Velence, Sukoró and Pákozd. To identify the sampled caterpillar, DNA analysis was undertaken. Additionally, in 2019–2020 a country-wide investigation was carried out to map the distribution and abundance of C. interscindana and gather data on bionomics of this pest in the Carpathian basin.

Transcriptome-wide Identification and Characterization of microRNAs and Their Targets in a Highly Adaptable Conifer Platycladus orientalis

MicroRNAs (miRNAs) are short noncoding RNAs (20–25 nucleotides) that regulate gene expression posttranscriptionally. However, identification and characterization of miRNAs remain limited for conifer species. In this study, we applied transcriptome-wide miRNAs sequencing to a conifer species Platycladus orientalis, which is highly adaptable to a wide range of environmental adversities, including drought, barren soil, and mild salinity. A total of 17,181,542 raw reads were obtained from the Illumina sequencing platform; 31 conserved and 91 novel miRNAs were identified, and their unique characteristics were further analyzed. Ten randomly selected miRNAs were validated by quantificational real-time polymerase chain reaction. Through miRNA target predictions based on psRNATarget, 2331 unique mRNAs were predicted to be targets of P. orientalis miRNAs that involved in 187 metabolic pathways in KEGG database. These targets included not only important transcription factors (e.g., class III homeodomain leucine zipper targeted by por-miR166d) but also indispensable nontranscriptional factor proteins (i.e., por-miR482a-3p regulated nucleotide-binding site leucine-rich repeat protein). Interestingly, six miRNAs (por-miR16, -miR44, -miR60-5p, -miR69–3p, -miR166b-5p, and -miR395c) were found in adaptation-related pathways (e.g., drought), indicating their possible involved in this species’ stress-tolerance characteristics. The present study provided essential information for understanding the regulatory role of miRNAs in P. orientalis and sheds light on their possible use in tree improvement for stress tolerance.

Plant extract as gas adsorbent to reduce smoke toxicants of cigarette

Cigarette smoke contains thousands of chemicals including many known toxicants and annually leads to millions of deaths worldwide. To reduce the harms of cigarette, plant extracts were applied to adsorb smoke toxicants of cigarette. Results showed that platycladus orientalis leaf extract and mulberry fruit extract particles filled into cigarette cellulose acetate filter can significantly reduce 15 major cigarette smoke toxicants emission including hydrogen cyanide (HCN), benzo[α] pyrene (B[a]P), formaldehyde, crotonaldehyde, 2-butanone, P-hydroquinone, M-dihydroxybenzene, catechol, phenol, M-P-cresol, O-cresol, N'-nitrosonornicotine (NNN), 4-methylnitrosamino-l-3-pyridyl- butanone (NNK), (R,S)-N-nitrosoanatabine (NAT) and (R,S)-N-nitrosoanabasine (NAB) by 11.90% to 60.42% (P<0.01). Platycladus orientalis leaf extract particles added in the outer cigarette filter also can adsorb other 125 kinds of chemicals most of which are harmful. Our results also indicated that plant extract has extensive gas adsorption characteristics and different plant extracts displayed different adsorption capacity to different toxicants. The adsorption capacities of five randomly selected plant extracts are all significantly higher than that of activated carbon（P<0.01). These findings suggest that plant extracts are excellent cigarette smoke adsorbents.

Transcriptional profiles of a foliar fungal endophyte (Pestalotiopsis, Ascomycota) and its endohyphal bacterium (Luteibacter, Gammaproteobacteria) in co-culture support sulfur exchange and growth regulation

Symbiosis with bacteria is widespread among eukaryotes, including fungi. Bacteria that live within fungal mycelia (endohyphal bacteria) occur in many plant-associated fungi, including diverse Mucoromycota and Dikarya. Pestalotiopsis sp. 9143 is a filamentous ascomycete isolated originally as a foliar endophyte of Platycladus orientalis (Cupressaceae). It is infected naturally with the endohyphal bacterium Luteibacter sp. 9143, which influences auxin and enzyme production by its fungal host. Previous studies have used transcriptomics to examine similar symbioses between endohyphal bacteria and root-associated fungi such as arbuscular mycorrhizal fungi and plant pathogens. However, currently there are no gene expression studies of endohyphal bacteria of Ascomycota, the most species-rich fungal phylum. We developed methods for assessing gene expression by Pestalotiopsis sp. and Luteibacter sp. when grown in co-culture and when each was grown axenically. Our assays showed that the density of Luteibacter sp. in co-culture was greater than in axenic culture, but the opposite was true for the Pestalotiopsis sp. Dual RNA-seq data demonstrate that growing in co-culture modulates developmental and metabolic processes in both the fungus and bacterium, potentially through changes in the balance of organic sulfur via methionine acquisition. Our analyses also suggest an unexpected, potential role of the bacterial type VI secretion system in symbiosis establishment, expanding current understanding of the scope and dynamics of fungal-bacterial symbioses.

Genetic Evaluation of Ancient Platycladus orientalis L. (Cupressaceae) in the Middle Reaches of the Yellow River Using Nuclear Microsatellite Markers

As a precious and rare genetic resource, ancient Platycladus orientalis L. (Cupressaceae) has important scientific, cultural and historical value. The ancient temples and royal cemeteries in the middle reaches of the Yellow River contain the most concentrated and abundant distributions of ancient P. orientalis. Due to unfavorable conditions, the genetic resources of ancient trees are facing great threats and challenges; thus, it is urgent to strengthen the evaluation of the genetic resources of ancient P. orientalis. In this study, we used nine polymorphic nuclear simple sequence repeats (nSSRs) to evaluate the genetic resources of 221 individuals in 19 ancient P. orientalis populations in the middle reaches of the Yellow River. These selected polymorphic nSSR loci can be used reliably and rapidly in P. orientalis genetic studies. Our study showed that the 19 ancient P. orientalis populations have high genetic diversity (mean H = 0.562, He = 0.377). High historical gene flow (mean Nm = 1.179) and high genetic differentiation (mean Fst = 0.184) were observed in the ancient P. orientalis population. The analysis of molecular variance (AMOVA) showed that higher genetic variation existed within populations (93%) rather than among populations (7%). The genetic structures showed that the 19 populations were divided into two groups. The Mantel test and neighbor-joining (NJ) tree analysis showed no geographical distribution characteristics among populations, which may indicate a history of transplanting by ancient humans. Our research provides a theoretical basis for the protection and utilization of ancient P. orientalis germplasm resources and exploration of the historical origin and genetic relationships among the populations of P. orientalis on a large scale in the future.

Partitioning tree water usage into storage and transpiration in a mixed forest

Background Water migration and use are important processes in trees. However, it is possible to overestimate transpiration by equating the water absorbed through the plant roots to that diffused back to the atmosphere through stomatal transpiration. Therefore, it is necessary to quantify the water transpired and stored in plants. Method The δ2H/δ18O technique and heat ratio method were used to explore the water usage of coniferous and broad-leaved tree species, including the proportions of water used for transpiration and water storage. Results Platycladus orientalis and Quercus variabilis had strong plasticity in their water usage from different sources. Platycladus orientalis primarily used groundwater (30.5%) and the 60–100-cm soil layer (21.6%) throughout the experimental period and was sensitive to precipitation, absorbing water from the 0–20-cm layer (26.6%) during the rainy season. Quercus variabilis absorbed water from all sources (15.7%–36.5%) except from the 40–60-cm soil layer during the dry season. In addition, it did not change its water source but increased its groundwater uptake during the rainy season. The annual mean water fluxes of P. orientalis and Q. variabilis were 374.69 and 469.50 mm·year− 1, with 93.49% and 93.91% of the water used for transpiration, respectively. However, nocturnal sap flow in P. orientalis and Q. variabilis was mainly used for water storage in the trunk rather than transpiration, which effectively alleviated drought stress and facilitated the transport of nutrients. Conclusions The water stored in both species comprised 6%–7% of the total water fluxes and, therefore, should be considered in water balance models.

Quantitative accuracy assessment of the revised sparse Gash model using distinct time-step climatic parameters

Rainfall interception (I) can considerably influence the transport process of water. The revised sparse Gash model (RSGM) is a tool for determining the I, which assumes that the two climate parameters in the model are equal for all storms. However, few studies have provided additional cases to reexamine the correctness of this assumption and investigated the response of I of single storms to the time-step variability in climatic parameters. Hence, running the RSGM separately on an event basis during the growing season in 2017 and using three time-step climatic parameters (storm-based, monthly, and fixed) to estimate I for the forest stands of Pinus tabuliformis, Platycladus orientalis, and Acer truncatum in Northern China. In summary, the modeling accuracy of both cumulative I and individual I was enhanced by increasing the time step of the climatic parameters in this study. These positively support the assumption in the RSGM. These results suggest that it is more appropriate to run the RSGM using fixed climate parameters to estimate I for these tree species during the growing season in northern China. Additionally, the assumption in the RSGM should be appealed to be further confirmed across the widest possible range of species, regions, and time scales.