The destructive invasive pathogen Phytophthora lateralis found on Chamaecyparis lawsoniana across the UK

In 2010–2011, Phytophthora lateralis was isolated from diseased Chamaecyparis lawsoniana exhibiting dieback and mortality at eight geographically separate forest, parkland and shelterbelt locations in England, Scotland and Northern Ireland. In 2011, P. lateralis was also isolated from young symptomatic nursery plants of C. lawsoniana and Thuja occidentalis recently imported into Scotland from mainland Europe. These are the first findings of P. lateralis in the UK. At six of the field sites, only collar and root lesions were observed. However, at two sites, large stem and branch lesions unconnected to the collar region were also observed. Phytophthora lateralis was readily isolated from both aerial and basal lesions. In artificial inoculation experiments, two Scottish isolates of the pathogen caused lesions on C. lawsoniana shoots and were readily reisolated from the lesions, their pathogenicity being comparable to that of P. lateralis isolates originating from outside the UK. Isolates from six field sites and the two nursery interceptions exhibited ITS and cox II sequences identical to published sequences of French and North American isolates. However, the isolates from two field sites shared an ITS sequence with Taiwanese isolates and differed from North American, French and Taiwanese isolates by a single‐base substitution in cox II, suggesting a separate evolutionary history. It is clear that P. lateralis now presents a significant threat to C. lawsoniana in Britain. The main source of the outbreaks is likely to be imported infested nursery stock.     

Genetic diversity of European populations of the oak fine‐root pathogen Phytophthora quercina

The recently discovered oak‐specific fine root plant pathogen Phytophthora quercina is a significant factor in the current phase of European oak decline but its origins and ecology are poorly understood. A genome‐wide analysis of 260 amplified fragment length polymorphism (AFLP) markers was used to examine the genetic diversity of 72 isolates from five oak species at 28 sites in Germany (particularly Bavaria), Italy, France, Hungary and the UK. Within‐site diversity was examined at 16 sites. The limited genetic diversity (within and between sites) and lack of genetic substructuring according to geographic origin or host species suggest the rapid spread of a relatively recently introduced species. Two subgroups were distinguished and these may reflect an initial introduction of isolates of two different genetic backgrounds. The relatively low genetic diversity is probably because of the predominantly inbreeding (homothallic) nature of P. quercina. However, evidence of limited intra‐site diversity, temporal variation and the lack of clonality within the European population suggest that some diversity is being maintained by occasional outcrossing and turnover of a reservoir of long‐lived soil‐borne oospore (sexually derived) inoculum.     

Land‐use changes influence the sporulation and survival of Phytophthora agathidicida,a lethal pathogen of New Zealand kauri (Agathis australis)

Phytophthora agathidicida is the accepted causal agent of dieback in remnant stands of long‐lived indigenous New Zealand kauri (Agathis australis) and poses a significant threat to the long‐term survival of this species. Little is known about the effect of key soil physicochemical characteristics on the growth of P. agathidicida. In this study, we investigated the growth of P. agathidicida in soils collected from adjacent areas under original kauri forest, short rotation pine (Pinus radiata) plantation forest and grazed pastures. A growth response assay was used to quantify asexual (sporangia) and sexual (oospore) spore counts over 8 days in soils sampled from each land‐use. Significantly higher numbers of sporangia (p < 0.001) and oospores (p < 0.01) were found in pasture and pine forest soil within 2 days of the growth assay trials, suggesting these soils may favour asexual/sexual reproduction in the early stages of P. agathidicida establishment compared to kauri forest soils. Additionally, oospore production significantly increased over 8 days in pine forest soil, suggesting that with an increase in inoculum loads, these soils potentially act as pathogen reservoirs. The soil physicochemical properties (e.g., pH, C and N, phosphorus content and electrical conductivity) investigated in this study did not significantly correspond to spore count data between land‐uses, suggesting that differences in growth response are driven by other edaphic factors not explored in the present study.     

Fast and specific detection of the invasive forest pathogen Heterobasidion irregulare through a Loop‐mediated isothermal AMPlification (LAMP) assay

Heterobasidion irregulare is one of the most destructive fungal pathogens of pines in North America and was accidentally introduced into central Italy, where it has become invasive. The fungus is currently recommended for regulation by the European and Mediterranean Plant Protection Organisation (EPPO). In this work, an efficient diagnostic tool for the early detection of H. irregulare based on Loop‐mediated isothermal AMPlification (LAMP) coupled with two different DNA extraction methods was developed. The LAMP assay showed high specificity and good sensitivity, with a limit of detection of about 20 picograms of target DNA and time of detection of less than 40 min. The assay was successfully tested on a variety of different samples, including fungal fruiting bodies, infected plants and colonized wood. A survey on environmental samples collected in the field was also performed using the LAMP assay coupled with a rapid DNA extraction method. The possible applications of this molecular diagnostic tool encompass the monitoring of pine forests surrounding the current invasion area, laboratory or in‐field analyses of samples from suspected trees, and the surveillance in the ports of entry of wood imported from North America.     

Estimation of the fine root biomass in a Japanese cedar (Cryptomeria japonica) plantation using minirhizotrons

We estimated fine root biomass in a Japanese cedar (Cryptomeria japonica) plantation using a min-irhizotron technique. Since data obtained from minirhizo-trons are limited to the length and diameter of fine roots observed on minirhizotron tubes, data conversion is necessary to determine the fine root biomass per unit soil volume or unit stand area. We first examined the regression between diameter squared and weight per unit length of fine roots in soil core samples, and calculated the fine root biomass on minirhizotron tubes from their length and diameter. Then we determined conversion factors based on the ratio of the fine root biomass in soil core samples to that on minirhizotron tubes. We examined calculation methods, using a single conversion factor for total fine root biomass in the soil for depths of 0–40cm (Cal1), or using four conversion factors for fine roots in the soil at 10-cm intervals (Cal2). Cal1 overestimated fine root biomass in the lower soil or underestimated that in the upper soil, while fine root biomass calculated using Cal2 better matched that in soil core samples. These results suggest that minirhizotron data should be converted separately for different soil depths to better estimate fine root biomass.     

Development of a quantitative real‐time PCR assay for the detection of Phytophthora austrocedrae,an emerging pathogen in Britain

A TaqMan real‐time PCR assay was developed for Phytophthora austrocedrae, an emerging pathogen causing severe damage to juniper in Britain. The primers amplified DNA of the target pathogen down to 1 pg of extracted DNA, in both the presence and absence of host DNA, but did not amplify any of the non‐target Phytophthora and fungal species tested. The assay provides a useful tool for screening juniper populations for the disease.     

A TaqMan real‐time PCR assay for the detection of Phytophthora ‘taxon Agathis’ in soil,pathogen of Kauri in New Zealand

Kauri Agathis australis, an iconic tree of New Zealand, is under threat from an introduced disease‐causing pathogen provisionally named Phytophthora ‘taxon Agathis’ (referred to as PTA). This soilborne, Pythiaceous species belongs to the Chromista and causes a collar rot resulting in yellowing of the foliage and thinning of the canopy, which eventually causes death of the infected tree. The management and containment of this pathogen requires rapid and reliable detection in the soil. The current method for soil detection utilizes a soil bioassay involving lupin baits and soil flooding in a process that takes between ten and twenty days. We describe a real‐time PCR assay based on TaqMan chemistry for the specific detection of PTA, which targets the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA. This TaqMan real‐time PCR assay could be used with DNA extracted directly from bulk soil samples to enable rapid quantification of PTA within soil. The detection limit was 2 fg of PTA DNA from pure culture, or 20 fg in the presence of DNA extracted from soil. The assay was validated using soil samples taken from a PTA‐infested site and soil spiked with a known concentration of oospores. We conclude that the TaqMan real‐time PCR assay offers a more time‐efficient method for detection of PTA in soil than existing methods.     

Molecular characteristics and functional analysis of the β‐1,4‐endoglucanase Bm‐eng‐1 gene of Bursaphelenchus mucronatus (Nematoda: Aphelenchoididae)

The pine nematode Bursaphelenchus mucronatus has been associated with pine wilt in China. This work was intended to investigate the role of the pathogenicity‐related β‐1,4‐endoglucanase gene of B. mucronatus in the infection of pines. In this study, the full‐length cDNA of the Bm‐eng‐1 gene was cloned and characterized from a B. mucronatus isolate. Phylogenetic analysis suggested that the Bm‐eng‐1 gene of B. mucronatus may be acquired from fungi through horizontal gene transfer. The function of the gene was demonstrated using RNA interference. RNA interference indicated that Bm‐eng‐1 was involved in the dispersal, reproductive ability and pathogenicity of B. mucronatus. In situ hybridization showed that Bm‐eng‐1 was specifically expressed in the oesophageal gland of B. mucronatus. Furthermore, to investigate the relation between β‐1,4‐endoglucanase activities and virulence of different isolates, the β‐1,4‐endoglucanase activities and the expression levels of Bm‐eng‐1 were detected in six B. mucronatus isolates with different virulence. β‐1,4‐Endoglucanase activity was generally higher in pathogenic isolates than in non‐pathogenic isolates, and the Bm‐eng‐1 expression levels in different isolates were positively correlated with the β‐1,4‐endoglucanase activity. These findings suggested that β‐1,4‐endoglucanase plays important roles in the pathogenic process of B. mucronatus, and the differential expression of the gene may underlie the different activity levels of β‐1,4‐endoglucanase, subsequently influencing variations in virulence of B. mucronatus isolates.     

A thirteen‐year study on the impact of a severe canker disease of Corymbia calophylla,a keystone tree in Mediterranean‐type forests

Worldwide, forests and woodlands have shown progressive declines in health as a result of global environmental changes in combination with local anthropogenic drivers. This study examined the incidence and progression of a canker disease of marri (Corymbia calophylla) caused by the endemic fungal pathogen Quambalaria coyrecup at three paired forest and anthropogenically disturbed sites in the southwest of Western Australia over 13 years. At the time of plot establishment in 2001, cankers were present on trees at all six sites with 22.7% of the assessed trees cankered. By 2014, cankers had led to the death of 6.7% of the trees, and an additional 10.0% of the trees developed cankers during this time. A further 2.3% of trees died due to causes other than canker, resulting in a final figure of 65.0% of trees remaining alive and free of cankers for the duration of the survey period. Canker incidence was significantly greater on trees present at anthropogenically disturbed sites (along roadsides and in paddocks) than forest trees (35.3% increasing to 50.7%, 10.0% increasing to 14.7%, respectively). Trunk diameter at breast height, tree height and crown ratings were not correlated with canker presence. This long‐term study provides evidence of the increasing severity of this canker pathogen and the impact it is having on the survival of marri.     

Vegetation status on Nishi-jima Island (Ogasawara) before eradication of alien herbivore mammals: rapid expansion of an invasive alien tree, <Emphasis Type="Italic">Casuarina equisetifolia</Emphasis> (Casuarinaceae)

Insular ecosystems can be dramatically affected by alien species, and records of pre-eradication status are essential to evaluate the effects of eradicating alien species. Nishi-jima Island is a small island of the Ogasawara group on which the first program of complete eradication of alien herbivorous mammals (black rats and feral goats) will be conducted. After eradication, the government plans to conduct ecosystem restoration on the island. This paper reports the angiosperm flora and vegetation of Nishi-jima Island before eradication of the rats and goats, with the objective of aiding ecosystem management after the eradication. Our surveys indicate that vegetation cover by the alien tree, Casuarina equisetifolia has expanded compared with its distribution in a 1974 aerial photograph of the island. The predominant vegetation in 2006 was C. equisetifolia forests and Zoysia tenuifolia grasslands, with fragmented native tree vegetation. The flora of the island comprised 69 angiosperm species (50 indigenous species) of which 30% were endemic, far less than for the Ogasawara Islands as a whole (45%). However, several populations of endangered plants remain. To restore the native ecosystem on Nishi-jima, eradication of C. equisetifolia is important in addition to eradication of alien herbivorous mammals.     

Logging history (1820–2000) of a heavily exploited southern boreal forest landscape: Insights from sunken logs and forestry maps

Over the last two centuries, logging has caused major, but unquantified, compositional and structural changes in the southern portion of the North American boreal forest. In this study, we used a series of old forest inventory maps coupled with a new dendrochronological approach for analyzing timber floating histories in order to document the long-term transformation (1820–2000) of a southern boreal landscape (117 000 ha) in eastern Quebec, Canada, in response to logging practices. Landscape exploitation became increasingly severe throughout this time period. During the ninetieth century (1820–1900) of limited industrial capacity, selective logging targeted pine and spruce trees and excluded balsam fir, a much abundant species of the forest landscape. Logging intensity increased during the first half of the twentieth century, and targeted all conifer species including balsam fir. After 1975, dramatic changes occurred over the landscape in relation to clear-cutting practices, plantations, and salvage logging, which promoted the proliferation of regenerating areas and extensive plantations of the previously uncommon black spruce. Overall, logging disturbance resulted in an inversion in the forest matrix, from conifer to mixed and deciduous, and from old to regenerating stands, thus creating significant consequences on forest sustainability. If biodiversity conservation and sustainable forestry are to be management goals in such a heavily exploited forested landscape, then restoration strategies should be implemented in order to stop the divergence of the forests from their preindustrial conditions.     

Effects of simulated drought stress on the fine roots of Japanese cedar (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) in a plantation forest on the Kanto Plain,eastern Japan

Drought stress was simulated in a 28-year-old Japanese cedar plantation (Kanto Plain, Japan) between April and October 2004 by removing throughfall using rain shelters. Changes in fine-root parameters caused by this drought treatment were examined by sequential soil coring. Drought effects on fine roots were analyzed separately for particular soil depths (0–5, 5–15, and 15–25 cm) and root diameters (<1 and 1–2 mm). Generally, fine-root biomass and root tip numbers decreased by the drought treatment. Drought stress was most intense for fine roots in the topsoil and weakest for fine roots in the deepest soil layer. Fine roots less than 1 mm in diameter were affected more severely than 1- to 2-mm roots. The effect of drought treatment was most remarkable for the number of white root tips, which decreased to 17% of the control at the soil depth of 0–5 cm. These results suggest that white root tip is the most suitable indicator of drought stress. Simulated drought reduced production of fine roots less than 1 mm and 1–2 mm in diameter. Fine-root mortality was stimulated for roots less than 1 mm, but not for 1- to 2-mm roots. These results suggest that fine roots with larger diameters can survive drought stress at a level simulated in this study, but processes of fine-root production were inhibited regardless of the diameter classes. The duration of drought stress and phenology of fine roots should also be considered in diagnosing the effects of drought on fine-root parameters.     

Annual variation of pathogen occurrence and pathogen prevalence in<Emphasis Type="Italic"> Ips typographus</Emphasis> (Coleoptera,Scolytidae) from the BOKU University Forest Demonstration Centre

Four different pathogen species (Gregarina typographi, Chytridiopsis typographi, Unikaryon montanum and Menzbieria chalcographi) were found in adult Ips typographus from the Forest Demonstration Centre, BOKU University in the course of a 7-year period of investigation (1995–2001). Annual variations were found in both the spectrum and prevalence of pathogen species. Variations were found in the number of infected male and female beetles as well as the infections of beetles with different dates of emergence.     

Early growth of hinoki (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>) trees under different topography and edge aspects at a strip-clearcut site in Kyushu,Southern Japan

The positive and negative effects of the residual stand edge at a strip-clearcut site were examined on the initial growth of hinoki (Chamaecyparis obtusa) planted in areas with diverse topography. On the south edge, the gap light index was lower than on the north edge and strip center; however, the vapor pressure deficit was also lower than on the north edge and strip center, which resulted in lower physiological stress of hinoki on the south edge. Tree size on the south edge did not exceed that on the north edge and strip center. These results indicated that low light conditions due to residual trees negatively affected growth on the south edge even under the positive effect of microclimate alleviation. In valley, tree size in the second year after planting was smaller than on ridge and slope; however, tree growth during the following 2 years was higher in valley. Surface soil in valley was thinner with rockier substrates than on ridge and slope; that is, soil sedimentation type and substrates influenced the initial growth of hinoki with undeveloped roots, and after that, topography started to influence growth, probably because hinoki roots penetrated into deeper soil, which is influenced by the water gathering capacity of the valley. The initial growth of hinoki at the strip-clearcut site was predominantly affected by light rather than microclimate alleviation. This alleviation is expected to disappear when hinoki trees planted in the strip center grow enough to give additional shade to slow-growing edge hinoki.     

Strain analysis near the cutting edge in orthogonal cutting of hinoki (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>) using a digital image correlation method

A digital image correlation (DIC) method was utilized to measure strain distributed within approximately 0.5 mm of the cutting edge during slow-speed orthogonal cutting of air-dried hinoki (Chamaecyparis obtusa), to clarify the relationships of the strain distribution and cutting conditions, including cutting angle (\(\theta\)) and depth of cut (\(d\)). The strain was measured in 0.04 mm steps, and the measurable minimum strain was approximately 0.08%. Tensile strain of 3% or larger normal to the cutting direction, \({\varepsilon _y}\), tended to extend 0.2 mm or further ahead of the tool when \(\theta \leq 60^\circ\) and \(d \geq 0.1{\text{ mm}}\). This tensile \({\varepsilon _y}\) corresponded to the occurrence of the fore-split in Chip Type I. The tensile \({\varepsilon _y}\) detected along the path of the cutting edge decreased as \(\theta\) and/or \(d\) decreased. Positive shear strain, \({\gamma _{xy}}\), tended to be detected ahead of the tool in Type I. Negative \({\gamma _{xy}}\) tended to be detected ahead of the tool in Type II and III \(\left( {\theta \geq 70^\circ ,\,\,d \geq 0.05{\text{ mm}}} \right)\). These \({\gamma _{xy}}\) values were considered to be related to the elongation and shrinkage of the chip. The study confirmed the usability of the DIC method for the evaluation of cutting conditions and also to classify chip formation into chip types.     

Statistical analysis of the spatio‐temporal dynamics of rubber tree (Hevea brasiliensis) trunk phloem necrosis: no evidence of pathogen transmission

Trunk phloem necrosis (TPN) of Hevea brasiliensis is an irreversible syndrome of the phloem that spreads from the collar towards the tapping cut. It is responsible for the cessation of latex production, the main constraint in rubber plantations worldwide. Numerous investigations have been undertaken to understand the mechanisms of this disease. The apparent linear spread of TPN supported the initial hypothesis of a biotic causal agent for TPN. However, previous and recent aetiological analyses remained inconclusive and the pathogen hypothesis is tending to be abandoned. In this paper we present a complementary statistical analysis of spatio‐temporal epidemiological data collected in a rubber plantation in Côte d'Ivoire. There, four study plots were surveyed each year from 2000 to 2003, with tree‐by‐tree disease assessment. In two plots, the tapping knife was systematically disinfected with sodium hypochlorite to stop any virus or viroid transmission. Based on permutation tests, our analysis confirmed the aetiological results: there was no evidence for spread by pathogen transmission. However, the spatial structure of the disease was clear. These results strengthen the current alternative scenario of a multi‐factor physiological disease caused by an accumulation of exogenous and endogenous stresses. Spatial heterogeneity of the risk factors probably results in the presence of areas of stress that can explain the spatial patterns observed among the TPN cases. The final contribution of this study was confirmation of the curative effect of sodium hypochlorite in the earliest stages of the disease, thus opening the way for control of this disease.     

Modeling of a cylindrical laminated veneer lumber I: mechanical properties of hinoki (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>) and the reliability of a nonlinear finite elements model of a four-point bending test

The use of low-grade logs to build spirally wound laminated veneer lumber (LVL) has been studied and improved from the point of view of the gluing process, fiber orientation angle, and end joint of the LVL. The butt joint appears to be the fracture point when the column is submitted to a compressive or bending load. Owing to the complexity of cylindrical LVL, we used a finite element method to simulate the mechanical behavior of part of its wall. This part was small enough to be considered flat but was representative of the structure, especially in the area of the butt joint. This allowed us to test the validity of different settings of the parameters involved in the manufacturing process. To feed data for this model, we used the results established for the linear and nonlinear behavior of raw hinoki in Part I of this series of articles. We then used this numerical model to improve the quality of the butt joint by testing different settings of the joint. We show that reducing the butt joint gap under 0.5 mm, which requires only a few changes in the production line, provides an important increase in the modulus of upture and nonnegligible improvement of the modulus of elasticity compared to that for a ≥ 1 mm butt joint gap.     

Effects of discharge level on the load of dissolved and particulate components of stream nitrogen and phosphorus from a small afforested watershed of Japanese cypress (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>)

The behavior of dissolved and particulate components of stream nitrogen (N) and phosphorus (P) were measured for 2 years in a small mountainous watershed covered primarily with a plantation forest of Japanese cypress (Chamaecyparis obtusa). The load of dissolved N and P bore a consistent relationship to discharge while the load of particulate N and P varied by up to two orders of magnitude at a given discharge level. Most N was exported in a dissolved form (DN) while most P was exported in a particulate form (PP), which bears similarity to loads from agricultural watersheds. Owing to the different behaviors of DN and PP, changes in the total nitrogen (TN) load were primarily attributed to variations in discharge unlike changes in the total phosphorus (TP) load. High flow conditions, resulting from heavy rainfall, displayed PP release significantly larger than expected. The TP load in high flow conditions was severely underestimated using a regression equation expressed as a function of discharge, which was based on the weekly sampling data biased toward low flow conditions. In addition, the TN load during peak discharges in heavy rain events was underestimated by the regression equation because of unpredictable increases in the particulate component. Our study shows that the particulate component ratio determines whether discharge can explain changes in load regardless of chemical species. The results suggest that plantation forests in rainy regions can be a diffuse source of particulate nutrients depending on soil surface conditions.     

Results of a thinning study in a black stain (Leptographium wageneri)‐infested Jeffrey/ponderosa pine forest in northern California

Black stain root disease (BSRD) caused by Leptographium wageneri var. ponderosum (Verticicladiella wageneri) is a destructive disease in many Jeffrey pine (Pinus jeffreyi) and ponderosa pine (P. ponderosa) forests of the western United States. Overcrowding is a pervasive problem in these forests, unless alleviated by thinning. In an overcrowded BSRD‐infested second‐growth Jeffrey/ponderosa pine forest in northern California, a large study (16.2 hr; 40 acres) was designed to determine the impact of thinning to four levels—6.9 m²/ha (30 ft²/acre), 13.8 m²/ha (60 ft²/acre), and 20.7 m²/ha (90 ft²/acre) of basal area and a control with no thinning—on the incidence of BSRD‐caused mortality. Ten years after this experiment was initiated, results indicate that thinning to any of these levels lowered the incidence of BSRD‐caused mortality as compared with BSRD‐caused mortality in unthinned control plots.     

Efficiency of extracting microarthropods from the canopy litter in a Japanese cedar (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis> D. Don) plantation: a comparison between the washing and Tullgren methods

We compared the efficiency of washing versus the Tullgren method for extracting microarthropods from dead leaves and branches in the canopy of Cryptomeria japonica trees. Oribatida and Collembola were consistently the numerically dominant taxa of microarthropods, but the relative abundance differed between the two extraction methods. Oribatida accounted for more than 70% of all the microarthropods collected by the washing method but less than 30% by the Tullgren method. Collembola accounted for less than 10% collected by the washing method but about 60% by the Tullgren method. The density of Oribatida was higher when collected by the washing method than by the Tullgren method, and vice versa for the density of Collembola. Our results suggest that the washing method is appropriate for collecting Oribatida and other microarthropods, whereas the Tullgren method is better for collecting Collembola from the canopy litter of C. japonica trees.