A delayed response in phytohormone signaling and production contributes to pine susceptibility to Fusarium circinatum.

BACKGROUND: Fusarium circinatum is the causal agent of pine pitch canker disease, which affects Pinus species worldwide, causing significant economic and ecological losses. In Spain, two Pinus species are most affected by the pathogen; Pinus radiata is highly susceptible, while Pinus pinaster has shown moderate resistance. In F. circinatum-Pinus interactions, phytohormones are known to play a crucial role in plant defense. By comparing species with different degrees of susceptibility, we aimed to elucidate the fundamental mechanisms underlying resistance to the pathogen. For this purpose, we used an integrative approach by combining gene expression and metabolomic phytohormone analyses at 5 and 10 days post inoculation. RESULTS: Gene expression and metabolite phytohormone contents suggested that the moderate resistance of P. pinaster to F. circinatum is determined by the induction of phytohormone signaling and hormone rearrangement beginning at 5 dpi, when symptoms are still not visible. Jasmonic acid was the hormone that showed the greatest increase by 5 dpi, together with the active gibberellic acid 4 and the cytokinin dehydrozeatin; there was also an increase in abscisic acid and salicylic acid by 10 dpi. In contrast, P. radiata hormonal changes were delayed until 10 dpi, when symptoms were already visible; however, this increase was not as high as that in P. pinaster. Indeed, in P. radiata, no differences in jasmonic acid or salicylic acid production were found. Gene expression analysis supported the hormonal data, since the activation of genes related to phytohormone synthesis was observed earlier in P. pinaster than in the susceptible P. radiata. CONCLUSIONS: We determine that the moderate resistance of P. pinaster to F. circinatum is in part a result of early and strong activation of plant phytohormone-based defense responses before symptoms become visible. We suggest that jasmonic acid signaling and production are strongly associated with F. circinatum resistance. In contrast, P. radiata susceptibility was attributed to a delayed response to the fungus at the moment when symptoms were visible. Our results contribute to a better understanding of the phytohormone-based defense mechanism involved in the Pinus-F. circinatum interactions and provide insight into the development of new strategies for disease mitigation.

Optimal Rotation Age in Fast Growing Plantations: A Dynamical Optimization Problem.

Forest plantations are economically and environmentally relevant, as they play a key role in timber production and carbon capture. It is expected that the future climate change scenario affects forest growth and modify the rotation age for timber production. However, mathematical models on the effect of climate change on the rotation age for timber production remain still limited. We aim to determine the optimal rotation age that maximizes the net economic benefit of timber volume in a negative scenario from the climatic point of view. For this purpose, a bioeconomic optimal control problem was formulated from a system of Ordinary Differential Equations (ODEs) governed by the state variables live biomass volume, intrinsic growth rate, and area affected by fire. Then, four control variables were associated to the system, representing forest management activities, which are felling, thinning, reforestation, and fire prevention. The existence of optimal control solutions was demonstrated, and the solutions of the optimal control problem were also characterized using Pontryagin's Maximum Principle. The solutions of the model were approximated numerically by the Forward-Backward Sweep method. To validate the model, two scenarios were considered: a realistic scenario that represents current forestry activities for the exotic species Pinus radiata D. Don, and a pessimistic scenario, which considers environmental conditions conducive to a higher occurrence of forest fires. The optimal solution that maximizes the net benefit of timber volume consists of a strategy that considers all four control variables simultaneously. For felling and thinning, regardless of the scenario considered, the optimal strategy is to spend on both activities depending on the amount of biomass in the field. Similarly, for reforestation, the optimal strategy is to spend as the forest is harvested. In the case of fire prevention, in the realistic scenario, the optimal strategy consists of reducing the expenses in fire prevention because the incidence of fires is lower, whereas in the pessimistic scenario, the opposite is true. It is concluded that the optimal rotation age that maximizes the net economic benefit of timber volume in P. radiata plantations is 24 and 19 years for the realistic and pessimistic scenarios, respectively. This corroborates that the presence of fires influences the determination of the optimal rotation age, and as a consequence, the net economic benefit.

Quantifying carbon storage and sequestration by native and non-native forests under contrasting climate types.

Non-native trees may have significant impacts on the carbon sink capacity of forested lands. However, large-scale patterns of the relative capacity of native and non-native forests to uptake and store carbon remain poorly described in the literature, and this information is urgently needed to support management decisions. In this study, we analyzed 17,065 plots from the Spanish Forest Inventory (covering c. 30 years) to quantify carbon storage and sequestration of natural forests and plantations of native and non-native trees under contrasting climate types, while controlling for the effects of environmental factors (forest structure, climate, soil, topography, and management). We found that forest origin (non-native vs. native) highly influenced carbon storage and sequestration, but such effect was dependent on climate. Carbon storage was greater in non-native than in native forests in both wet and dry climates. Non-native forests also had greater carbon sequestration than native ones in the wet climate, due to higher carbon gains by tree growth. However, in the dry climate, native forests had greater carbon gains by tree ingrowth and lower carbon loss by tree mortality than non-native ones. Furthermore, forest type (classified by the dominant species) and natural forests versus tree plantations were important determinants of carbon storage and sequestration. Native and non-native Pinus spp. forests had low carbon storage, whereas non-native Eucalyptus spp. forests and native Quercus spp., Fagus sylvatica, and Eurosiberian mixed forests (especially not planted ones) had high carbon storage. Carbon sequestration was greatest in Eucalyptus globulus, Quercus ilex, and Pinus pinaster forests. Overall, our findings suggest that the relative capacity of native and non-native forests to uptake and store carbon depends on climate, and that the superiority of non-native forests over native ones in terms of carbon sequestration declines as the abiotic filters become stronger (i.e., lower water availability and higher climate seasonality).

Analysis of the transcriptome of the needles and bark of Pinus radiata induced by bark stripping and methyl jasmonate.

BACKGROUND: Plants are attacked by diverse insect and mammalian herbivores and respond with different physical and chemical defences. Transcriptional changes underlie these phenotypic changes. Simulated herbivory has been used to study the transcriptional and other early regulation events of these plant responses. In this study, constitutive and induced transcriptional responses to artificial bark stripping are compared in the needles and the bark of Pinus radiata to the responses from application of the plant stressor, methyl jasmonate. The time progression of the responses was assessed over a 4-week period. RESULTS: Of the 6312 unique transcripts studied, 86.6% were differentially expressed between the needles and the bark prior to treatment. The most abundant constitutive transcripts were related to defence and photosynthesis and their expression did not differ between the needles and the bark. While no differential expression of transcripts were detected in the needles following bark stripping, in the bark this treatment caused an up-regulation and down-regulation of genes associated with primary and secondary metabolism. Methyl jasmonate treatment caused differential expression of transcripts in both the bark and the needles, with individual genes related to primary metabolism more responsive than those associated with secondary metabolism. The up-regulation of genes related to sugar break-down and the repression of genes related with photosynthesis, following both treatments was consistent with the strong down-regulation of sugars that has been observed in the same population. Relative to the control, the treatments caused a differential expression of genes involved in signalling, photosynthesis, carbohydrate and lipid metabolism as well as defence and water stress. However, non-overlapping transcripts were detected between the needles and the bark, between treatments and at different times of assessment. Methyl jasmonate induced more transcriptional responses in the bark than bark stripping, although the peak of expression following both treatments was detected 7 days post treatment application. The effects of bark stripping were localised, and no systemic changes were detected in the needles. CONCLUSION: There are constitutive and induced differences in the needle and bark transcriptome of Pinus radiata. Some expression responses to bark stripping may differ from other biotic and abiotic stresses, which contributes to the understanding of plant molecular responses to diverse stresses. Whether the gene expression changes are heritable and how they differ between resistant and susceptible families identified in earlier studies needs further investigation.

Genome-wide identification and characterization of Fusarium circinatum-responsive lncRNAs in Pinus radiata.

BACKGROUND: One of the most promising strategies of Pine Pitch Canker (PPC) management is the use of reproductive plant material resistant to the disease. Understanding the complexity of plant transcriptome that underlies the defence to the causal agent Fusarium circinatum, would greatly facilitate the development of an accurate breeding program. Long non-coding RNAs (lncRNAs) are emerging as important transcriptional regulators under biotic stresses in plants. However, to date, characterization of lncRNAs in conifer trees has not been reported. In this study, transcriptomic identification of lncRNAs was carried out using strand-specific paired-end RNA sequencing, from Pinus radiata samples inoculated with F. circinatum at an early stage of infection. RESULTS: Overall, 13,312 lncRNAs were predicted through a bioinformatics approach, including long intergenic non-coding RNAs (92.3%), antisense lncRNAs (3.3%) and intronic lncRNAs (2.9%). Compared with protein-coding RNAs, pine lncRNAs are shorter, have lower expression, lower GC content and harbour fewer and shorter exons. A total of 164 differentially expressed (DE) lncRNAs were identified in response to F. circinatum infection in the inoculated versus mock-inoculated P. radiata seedlings. The predicted cis-regulated target genes of these pathogen-responsive lncRNAs were related to defence mechanisms such as kinase activity, phytohormone regulation, and cell wall reinforcement. Co-expression network analysis of DE lncRNAs, DE protein-coding RNAs and lncRNA target genes also indicated a potential network regulating pectinesterase activity and cell wall remodelling. CONCLUSIONS: This study presents the first comprehensive genome-wide analysis of P. radiata lncRNAs and provides the basis for future functional characterizations of lncRNAs in relation to pine defence responses against F. circinatum.

Effects of climate change on forest plantation productivity in Chile.

Forest plantations in Chile occupy more than 2.2 million ha and are responsible for 2.1% of the GDP of the country's economy. The ability to accurately predictions of plantations productivity under current and future climate has an impact can enhance on forest management and industrial wood production. The use of process-based models to predict forest growth has been instrumental in improving the understanding and quantifying the effects of climate variability, climate change, and the impact of atmospheric CO2 concentration and management practices on forest growth. This study uses the 3-PG model to predict future forest productivity Eucalyptus globulus and Pinus radiata. The study integrates climate data from global circulation models used in CMIP5 for scenarios RCP26 and RCP85, digital soil maps for physical and chemical variables. Temporal and spatial tree growth inventories were used to compare with the 3-PG predictions. The results indicated that forest productivity is predicted to potentially increase stand volume (SV) over the next 50 years by 26% and 24% for the RCP26 scenario and between 73% and 62% for the RCP85 scenario for E. globulus and P. radiata, respectively. The predicted increases can be explained by a combination of higher level of atmospheric CO2 , air temperatures closer to optimum than current, and increases in tree water use efficiency. If the effect of CO2 is not considered, the predicted differences of SV for 2070 are 16% and 14% for the RCP26 scenario and 22% and 14% for RCP85 for the two species. While shifts in climate and increasing CO2 are likely to benefit promote higher productivity, other factors such as lack insufficient availability of soil nutrients, events such as increasing frequency and duration of droughts, longer periods of extreme temperatures, competing vegetation, and occurrence of new pests and diseases may compromise these potential gains.

Chemical Traits that Predict Susceptibility of Pinus radiata to Marsupial Bark Stripping.

Bark stripping by mammals is a major problem in managed conifer forests worldwide. In Australia, bark stripping in the exotic plantations of Pinus radiata is mainly caused by native marsupials and results in reduced survival, growth, and in extreme cases death of trees. Herbivory is influenced by a balance between primary metabolites that are sources of nutrition and secondary metabolites that act as defences. Identifying the compounds that influence herbivory may be a useful tool in the management of forest systems. This study aimed to detect and identify both constitutive and induced compounds that are associated with genetic differences in susceptibility of two-year-old P. radiata trees to bark stripping by marsupials. An untargeted profiling of 83 primary and secondary compounds of the needles and bark samples from 21 susceptible and 21 resistant families was undertaken. These were among the most and least damaged families, respectively, screened in a trial of 74 families that were exposed to natural field bark stripping by marsupials. Experimental plants were in the same field trial but protected from bark stripping and a subset were subjected to artificial bark stripping to examine induced and constitutive chemistry differences between resistant and susceptible families. Machine learning (random forest), partial least squares plus discriminant analysis (PLS-DA), and principal components analysis with discriminant analysis (PCA-DA), as well as univariate methods were used to identify the most important totals by compound group and individual compounds differentiating the resistant and susceptible families. In the bark, the constitutive amount of two sesquiterpenoids - bicyclogermacrene and an unknown sesquiterpenoid alcohol -were shown to be of higher levels in the resistant families, whereas the constitutive sugars, fructose, and glucose, as well individual phenolics, were higher in the more susceptible families. The chemistry of the needles was not useful in differentiating the resistant and susceptible families to marsupial bark stripping. After artificial bark stripping, the terpenes, sugars, and phenolics responded in both the resistant and susceptible families by increasing or reducing amounts, which leveled the differences in the amounts of the compounds between the different resistant and susceptible classes observed at the constitutive level. Overall, based on the families with extreme values for less and more susceptibility, differences in the amounts of secondary compounds were subtle and susceptibility due to sugars may outweigh defence as the cause of the genetic variation in bark stripping observed in this non-native tree herbivory system.

Hydroxypropylation of Polyphenol-Rich Alkaline Extracts from Pinus radiata Bark and Their Physicochemical Properties.

Pinus radiata bark is a rich source of polyphenols, which are mainly composed of proanthocyanidins. This study aimed to utilize P. radiata bark as a polyol source for bio-foam production in the future. Polyphenol-rich alkaline extracts (AEs) from P. radiata bark were prepared by mild alkaline treatment and then derivatized with propylene oxide (PO). Hydroxypropylated alkaline extracts (HAEs) with varying molar substitutions (MS 0.4-8.0) were characterized by FT-IR, NMR, GPC, TGA, and DSC. The hydroxyl value and solubility in commercial polyols were also determined. The molecular weights of the acetylated HAEs (Ac-HAEs) were found to be 4000 to 4900 Da. Analyses of FT-IR of HAEs and 1H NMR of Ac-HAEs indicated that the aromatic hydroxyl groups were hydroxypropylated and showed an increase in aliphatic hydroxyl group content. The glass transition temperature (Tg) of AE and HAEs were 58 to 60 °C, showing little difference. The hydroxyl value increased as the hydroxypropylation proceeded. Although salts were produced upon neutralization after hydroxypropylation, HAEs still showed suitable solubility in polyether and polyester polyols; HAEs dissolved well in polyether polyol, PEG#400, and solubility reached about 50% (w/w). This indicated that neutralized HAEs could be directly applied to bio-foam production even without removing salts.

Genome editing with CRISPR/Cas9 in Pinus radiata (D. Don).

BACKGROUND: To meet increasing demand for forest-based products and protect natural forests from further deforestation requires increased productivity from planted forests. Genetic improvement of conifers by traditional breeding is time consuming due to the long juvenile phase and genome complexity. Genetic modification (GM) offers the opportunity to make transformational changes in shorter time frames but is challenged by current genetically modified organism (GMO) regulations. Genome editing, which can be used to generate site-specific mutations, offers the opportunity to rapidly implement targeted improvements and is globally regulated in a less restrictive way than GM technologies. RESULTS: We have demonstrated CRISPR/Cas9 genome editing in P. radiata targeting a single-copy cell wall gene GUX1 in somatic embryogenic tissue and produced plantlets from the edited tissue. We generated biallelic INDELs with an efficiency of 15 % using a single gRNA. 12 % of the transgenic embryogenic tissue was edited when two gRNAs were used and deletions of up to 1.3 kb were identified. However, the regenerated plants did not contain large deletions but had single nucleotide insertions at one of the target sites. We assessed the use of CRISPR/Cas9 ribonucleoproteins (RNPs) for their ability to accomplish DNA-free genome editing in P. radiata. We chose a hybrid approach, with RNPs co-delivered with a plasmid-based selectable marker. A two-gRNA strategy was used which produced an editing efficiency of 33 %, and generated INDELs, including large deletions. Using the RNP approach, deletions found in embryogenic tissue were also present in the plantlets. But, all plants produced using the RNP strategy were monoallelic. CONCLUSIONS: We have demonstrated the generation of biallelic and monoallelic INDELs in the coniferous tree P. radiata with the CRISPR/Cas9 system using plasmid expressed Cas9 gRNA and RNPs respectively. This opens the opportunity to apply genome editing in conifers to rapidly modify key traits of interest.

Interclonal variation, coordination, and trade-offs between hydraulic conductance and gas exchange in Pinus radiata: consequences on plant growth and wood density.

Stem growth reflects genetic and phenotypic differences within a tree species. The plant hydraulic system regulates the carbon economy, and therefore variations in growth and wood density. A whole-organism perspective, by partitioning the hydraulic system, is crucial for understanding the physical and physiological processes that coordinately mediate plant growth. The aim of this study was to determine whether the relationships and trade-offs between (i) hydraulic traits and their relative contribution to the whole-plant hydraulic system, (ii) plant water transport, (iii) CO2 assimilation, (iv) plant growth, and (v) wood density are revealed at the interclonal level within a variable population of 10 Pinus radiata (D. Don) clones for these characters. We demonstrated a strong coordination between several plant organs regarding their hydraulic efficiency. Hydraulic efficiency, gas exchange, and plant growth were intimately linked. Small reductions in stem wood density were related to a large increase in sapwood hydraulic efficiency, and thus to plant growth. However, stem growth rate was negatively related to wood density. We discuss insights explaining the relationships and trade-offs of the plant traits examined in this study. These insights provide a better understanding of the existing coordination, likely to be dependent on genetics, between the biophysical structure of wood, plant growth, hydraulic partitioning, and physiological plant functions in P. radiata.

Tolerance of 2-Benzoxazolinone and Interactions with Grass and Pine Hosts in a Population of Fusarium circinatum.

Fusarium circinatum, the causal agent of pitch canker in pines and a cryptic endophyte of grasses, was examined for heritable variation in tolerance of the grass defense compound 2-benzoxazolinone (BOA). A diverse population of F. circinatum progeny was assayed for growth rate on potato dextrose agar amended with BOA. Matings were conducted to allow for selection of progeny with lower and higher tolerance of BOA. The results confirmed heritable variation in BOA tolerance in F. circinatum. A subset of differentially tolerant progeny was used for inoculations of growth chamber-grown Zea mays and greenhouse-grown Pinus radiata. No differences were detected in the rate of infection or extent of colonization of Z. mays inoculated with F. circinatum progeny differing in tolerance of BOA. Pitch canker symptoms in inoculated P. radiata trees showed that high BOA-tolerating isolates induced significantly longer lesion lengths than those induced by low BOA-tolerating isolates. Results from this study were consistent with the proposition that F. circinatum evolved from grass-colonizing ancestors and that pathogenicity to pine is a relatively recent evolutionary innovation.

Dual RNA-Sequencing Analysis of Resistant (Pinus pinea) and Susceptible (Pinus radiata) Hosts during Fusarium circinatum Challenge.

Fusarium circinatum causes one of the most important diseases of conifers worldwide, the pine pitch canker (PPC). However, no effective field intervention measures aiming to control or eradicate PPC are available. Due to the variation in host genetic resistance, the development of resistant varieties is postulated as a viable and promising strategy. By using an integrated approach, this study aimed to identify differences in the molecular responses and physiological traits of the highly susceptible Pinus radiata and the highly resistant Pinus pinea to F. circinatum at an early stage of infection. Dual RNA-Seq analysis also allowed to evaluate pathogen behavior when infecting each pine species. No significant changes in the physiological analysis were found upon pathogen infection, although transcriptional reprogramming was observed mainly in the resistant species. The transcriptome profiling of P. pinea revealed an early perception of the pathogen infection together with a strong and coordinated defense activation through the reinforcement and lignification of the cell wall, the antioxidant activity, the induction of PR genes, and the biosynthesis of defense hormones. On the contrary, P. radiata had a weaker response, possibly due to impaired perception of the fungal infection that led to a reduced downstream defense signaling. Fusarium circinatum showed a different transcriptomic profile depending on the pine species being infected. While in P. pinea, the pathogen focused on the degradation of plant cell walls, active uptake of the plant nutrients was showed in P. radiata. These findings present useful knowledge for the development of breeding programs to manage PPC.

Trade-offs between environmental and economic factors in conversion from exotic pine production to natural regeneration on erosion prone land.

Background: Some of New Zealand's exotic pine (Pinus radiata D.Don) forests were planted for erosion mitigation but cultural, legislative, environmental, and profitability limitations in some parts of the landscape have led to reassessment of their suitability. There is limited information to support landowner decisions on the viability of natural regeneration of native forest post-pine-harvest. Methods: We evaluated scenarios of post-harvest natural regeneration, compared to remaining in pine production, using erosion susceptibility determined from historical occurrence of landslides, gullies and earthflows, biophysical growth modelling of manuka-kanuka (Leptospermum scoparium-Kunzea ericoides (A.Rich) Joy Thomps.) shrubland using the process-based CenW model, and cost-benefit analyses using NZFARM with two land use change scenarios, at two levels of erosion mitigation ± honey profits. Results: In our study area, the Gisborne Region (North Island of New Zealand), ~27% of the land has moderate-very high susceptibility to landslides, 14-22% a high probability of contributing material to waterways, and 19% moderate-very high gully erosion susceptibility. Pines grow 10 times faster than naturally regenerating manuka-kanuka shrubland, but manuka-kanuka is used for honey not wood production. Natural regeneration resulted in losses of $150-250 ha-1 yr-1 compared to the current profitability of pine production. Honey production offset some reduction in pine revenue, but not fully. Thus, the viability of shifting from pines to native forest is highly dependent on landowner impetus and value for non-market ecosystem services (such as cultural and biodiversity values) provided by native forest. Conclusions: A mosaic of land uses within a property may sufficiently offset income losses with other benefits, whereby highly erosion-prone land is shifted from rotational pine forest production to permanent native forest cover with honey production where possible. At the regional scale in Gisborne, the conversion of the most highly susceptible land under production forestry (315-556 ha) to natural regeneration has the potential for wider benefits for soil conservation reducing erosion by 1-2.5 t yr-1 of sediment facilitating achievement of cleaner water aspirations and habitat provision.

Effect of trait's expression level on single-step genomic evaluation of resistance to Dothistroma needle blight.

BACKGROUND: Many conifer breeding programs are paying increasing attention to breeding for resistance to needle disease due to the increasing importance of climate change. Phenotyping of traits related to resistance has many biological and temporal constraints that can often confound the ability to achieve reliable phenotypes and consequently, reliable genetic progress. The development of next generation sequencing platforms has also enabled implementation of genomic approaches in species lacking robust reference genomes. Genomic selection is, therefore, a promising strategy to overcome the constraints of needle disease phenotyping. RESULTS: We found high accuracy in the prediction of genomic breeding values in the disease-related traits that were well characterized, reaching 0.975 for genotyped individuals and 0.587 for non-genotyped individuals. This compared well with pedigree-based accuracies of up to 0.746. Surprisingly, poorly phenotyped disease traits also showed very high accuracy in terms of correlation of predicted genomic breeding values with pedigree-based counterparts. However, this was likely caused by the fact that both were clustered around the population mean, while deviations from the population mean caused by genetic effects did not appear to be well described. Caution should therefore be taken with the interpretation of results in poorly phenotyped traits. CONCLUSIONS: Implementation of genomic selection in this test population of Pinus radiata resulted in a relatively high prediction accuracy of needle loss due to Dothistroma septosporum compared with a pedigree-based approach. Using genomics to avoid biological/temporal constraints where phenotyping is reliable appears promising. Unsurprisingly, reliable phenotyping, resulting in good heritability estimates, is a fundamental requirement for the development of a reliable prediction model. Furthermore, our results are also specific to the single pathogen mating-type that is present in New Zealand, and may change with future incursion of other pathogen varieties. There is no doubt, however, that once a robust genomic prediction model is built, it will be invaluable to not only select for host tolerance, but for other economically important traits simultaneously. This tool will thus future-proof our forests by mitigating the risk of disease outbreaks induced by future changes in climate.

No carbon limitation after lower crown loss in Pinus radiata.

BACKGROUND AND AIMS: Biotic and abiotic stressors can cause different defoliation patterns within trees. Foliar pathogens of conifers commonly prefer older needles and infection with defoliation that progresses from the bottom crown to the top. The functional role of the lower crown of trees is a key question to address the impact of defoliation caused by foliar pathogens. METHODS: A 2 year artificial defoliation experiment was performed using two genotypes of grafted Pinus radiata to investigate the effects of lower-crown defoliation on carbon (C) assimilation and allocation. Grafts received one of the following treatments in consecutive years: control-control, control-defoliated, defoliated-control and defoliated-defoliated. RESULTS: No upregulation of photosynthesis either biochemically or through stomatal control was observed in response to defoliation. The root:shoot ratio and leaf mass were not affected by any treatment, suggesting prioritization of crown regrowth following defoliation. In genotype B, defoliation appeared to impose C shortage and caused reduced above-ground growth and sugar storage in roots, while in genotype A, neither growth nor storage was altered. Root C storage in genotype B decreased only transiently and recovered over the second growing season. CONCLUSIONS: In genotype A, the contribution of the lower crown to the whole-tree C uptake appears to be negligible, presumably conferring resilience to foliar pathogens affecting the lower crown. Our results suggest that there is no C limitation after lower-crown defoliation in P. radiata grafts. Further, our findings imply genotype-specific defoliation tolerance in P. radiata.

Infection of the Native California Grass, Bromus carinatus, by Fusarium circinatum, the Cause of Pitch Canker in Pines.

At Point Reyes National Seashore in California, Fusarium circinatum, the causal agent of pitch canker in pines, was isolated from Pinus muricata, the California native grass, Bromus carinatus, and the introduced grass, Holcus lanatus. All grass plants from which F. circinatum was isolated were symptomless. Pathogenicity of grass isolates was confirmed by inoculation of P. radiata trees, which developed symptoms similar to trees inoculated with a pine isolate of F. circinatum. Isolates from grasses were somatically compatible with isolates recovered from symptomatic pines. B. carinatus grown in a growth chamber was inoculated with a green fluorescent protein-expressing strain of F. circinatum. Segments of inoculated leaves were incubated in moist chambers; after 1 to 2 days, sporulating hyphae were observed growing from leaf tissue. Spores of F. circinatum removed from B. carinatus leaves were confirmed to be fluorescent when illuminated with ultraviolet light. These results raise the possibility that B. carinatus cryptically infected by F. circinatum may be a source of propagules capable of infecting pines.

Wood Modification by Furfuryl Alcohol Resulted in a Delayed Decomposition Response in Rhodonia (Postia) placenta.

The aim of this study was to investigate differential expression profiles of the brown rot fungus Rhodonia placenta (previously Postia placenta) harvested at several time points when grown on radiata pine (Pinus radiata) and radiata pine with three different levels of modification by furfuryl alcohol, an environmentally benign commercial wood protection system. The entire gene expression pattern of a decay fungus was followed in untreated and modified wood from initial to advanced stages of decay. The results support the current model of a two-step decay mechanism, with the expression of genes related to initial oxidative depolymerization, followed by an accumulation of transcripts of genes related to the hydrolysis of cell wall polysaccharides. When the wood decay process is finished, the fungus goes into starvation mode after five weeks when grown on unmodified radiata pine wood. The pattern of repression of oxidative processes and oxalic acid synthesis found in radiata pine at later stages of decay is not mirrored for the high-furfurylation treatment. The high treatment level provided a more unpredictable expression pattern throughout the incubation period. Furfurylation does not seem to directly influence the expression of core plant cell wall-hydrolyzing enzymes, as a delayed and prolonged, but similar, pattern was observed in the radiata pine and the modified experiments. This indicates that the fungus starts a common decay process in the modified wood but proceeds at a slower pace as access to the plant cell wall polysaccharides is restricted. This is further supported by the downregulation of hydrolytic enzymes for the high treatment level at the last harvest point (mass loss, 14%). Moreover, the mass loss does not increase during the last weeks. Collectively, this indicates a potential threshold for lower mass loss for the high-furfurylation treatment.IMPORTANCE Fungi are important decomposers of woody biomass in natural habitats. Investigation of the mechanisms employed by decay fungi in their attempt to degrade wood is important for both the basic scientific understanding of ecology and carbon cycling in nature and for applied uses of woody materials. For wooden building materials, long service life and carbon storage are essential, but decay fungi are responsible for massive losses of wood in service. Thus, the optimization of durable wood products for the future is of major importance. In this study, we have investigated the fungal genetic response to furfurylated wood, a commercial environmentally benign wood modification approach that improves the service life of wood in outdoor applications. Our results show that there is a delayed wood decay by the fungus as a response to furfurylated wood, and new knowledge about the mechanisms behind the delay is provided.

First record of North American fungus Rhizopogon pseudoroseolus in Australia and prediction of its occurrence based on climatic niche and symbiotic partner preferences.

In 2017 a North American fungus, Rhizopogon pseudoroseolus (Boletales, Basidiomycota), formerly known in Oceania as only occurring in New Zealand, was found for the first time in South Australia. The morphological identification of collected specimens was confirmed using an internal transcribed spacer barcoding approach. In this study, the biogeography of R. pseudoroseolus is also presented, based on sporocarp and ectomycorrhiza records. Species distribution modeling implemented in MaxEnt was used to estimate the distribution of the potential range of R. pseudoroseolus in Australia and New Zealand. The obtained model illustrates, in the background of climatic variables and distribution of a symbiotic partner, its wide range of suitable habitats in New Zealand, South-East Australia, and Tasmania. Precipitation of the coldest quarters and annual mean temperature are important factors influencing the potential distribution of the fungus. The occurrence of Pinus radiata, the ectomycorrhizal partner of R. pseudoroseolus, is also an important factor limiting expansion of the fungus' invasion range.

Proanthocyanidin-rich Pinus radiata bark extract inhibits mast cell-mediated anaphylaxis-like reactions.

Mast cells play a critical role in the effector phase of immediate hypersensitivity and allergic reactions. Pinus radiata bark extract exerts multiple biological effects and exhibits immunomodulatory and antioxidant properties. However, its role in mast cell-mediated anaphylactic reactions has not been thoroughly investigated. In this study, we examined the effects of proanthocyanidin-rich water extract (PAWE) isolated from P. radiata bark on compound 48/80-induced or antidinitrophenyl (DNP) immunoglobulin E (IgE)-mediated anaphylaxis-like reactions in vivo. In addition, we evaluated the mechanism underlying the inhibitory effect of PAWE on mast cell activation, with a specific focus on histamine release, using rat peritoneal mast cells. PAWE attenuated compound 48/80-induced or anti-DNP IgE-mediated passive cutaneous anaphylaxis-like reactions in mice, and it inhibited histamine release triggered by compound 48/80, ionophore A23187, or anti-DNP IgE in rat peritoneal mast cells in vitro. Moreover, PAWE suppressed compound 48/80-elicited calcium uptake in a concentration-dependent manner and promoted a transient increase in intracellular cyclic adenosine-3',5'-monophosphate levels. Together, these results suggest that proanthocyanidin-rich P. radiata bark extract effectively inhibits anaphylaxis-like reactions.

Sex-biased secondary contact obscures ancient speciation onto relictual host trees in central California moths (Syndemis: Tortricidae).

The tortricid moth genus Syndemis has ten described species, with two polyphagous species in Europe and North America respectively. We sequenced five nuclear and four mitochondrial genes for Syndemis samples across both continents and discovered unexpected, extensive diversification restricted to California. DNA evidence supports five new, undescribed, species endemic to California, while the rest of North America and Europe have only one species each. Further, the nuclear genes are less variable and yield contrasting phylogenetic signal compared to mitochondrial DNA for basal relationships between taxa across the genus. Such conflict strongly suggests that male and female moths exhibit radically different levels of philopatry. Our results highlight the importance of sex-specific behavior, and the need for inclusion of multiple genes to fully understand species boundaries, their causes, and the process of speciation. While mtDNA introgression often is invoked to explain incongruous haplotype distributions, our study shows that nuclear DNA selective sweeps, or swamping, can occur while mtDNA and ecology preserve an ancient divergence that is not discernable in nuclear DNA. This study further demonstrates that diversification of herbivores may occur on relictual, declining hostplants, which contrasts with the dominant co-speciation model.