Biosafety assessment of transgenic poplars overexpressing xyloglucanase (AaXEG2) prior to field trials

We performed biosafety assessments of transgenic poplars prior to field trials. Constitutive expression of the Aspergillus aculeatus xyloglucanase in Populus alba increased the cellulose content and specific gravity of its stem, the leaves of which were visibly greener, thicker, and smaller than those of the wild-type plant. Although the young transgenic poplars grew faster than the wild type in a growth chamber, there was no distinguishable difference in growth between the poplars when they were placed in a special screened greenhouse. Allelopathic tests showed that the transgenic poplars do not produce harmful substances. Based on all the biosafety assessments and the scientific literature on poplar species, we came to the conclusion that transgenic poplars probably do not disturb the biological diversity of the surrounding environment, even when they are submitted to field trials.     

Photosynthesis and chlorophylla fluorescence of two poplars under water stress

In order to estimate drought tolerance in two species ofPopulus, Populus alba var. pyramidalis Bunge andPopulus nigra L. var. thevestina (Dode), widely planted at the southern margin of the Taklimakan Desert, responses of net photosynthesis and chlorophylla fluorescence to irradiance and water stress were examined under laboratory conditions. Results showed thatP. alba exhibited stronger drought tolerance thanP. nigra. A linear relationship between net photosynthetic rates (A _n) and electron transport rates (ETR) was found in both poplars under different irradiance and leaf water potentials. Net photosynthetic rates (A _n) in the two poplars significantly correlated linearly with the photochemical efficiency of the saturation light-adapted leaves throughout the range of leaf water potentials, suggesting that the leaf photochemical efficiency in saturation light-adapted leaves can be used to estimate leaf photosynthetic capacity and leaf water conditions in the two poplars within a magnitude of air temperature between 20 and 30°C. This research was supported by the Japan Society for the Promotion of Science (CAS-9601), the Chinese Academy of Sciences and partly supported by the National Natural Foundation of China (39870154).     

Effect of Melampsora medusae leaf rust infection on yield of hybrid poplars in the north-central United States

Melampsora medusae Thuem. is a hazard to cultivation and fiber production of hybrid poplars grown under short rotations, particularly when the alternate host is present. This rust can be controlled with biweekly fungicide applications but resistant clones are the best method of control.     

Inter‐ and intraspecific variation and genotype × site interaction in Salix Alba L., S. Dasyclados Wimm. and S. viminalis L.

Three test series were planted on sites selected for their contrasting properties to study the genotype × site interaction. Various clones of Salix alba (8 clones in the 1980 series), S. dasyclados (8 clones in 1982 series) and S. viminalis (6, 9, 13 clones in the various series) were included. Growth capacity, survival and infection by rust fungi were analysed. Interspecific variation was seen at some of the sites in the 1982 series but it was pronounced in the 1980 series where 5. viminalis outgrew 5. alba at all sites. Intraspecific variation could be seen for growth capacity, survival and rust resistance. A genotype × site interaction was detected for all variables studied. Some of the interaction, especially at the species level, can be explained by the variation in susceptibility to rust fungi and preference for soil types. In the breeding work with Salix the genotype × site interaction must be considered.     

Host range and pathogenicity studies of Marssonina castagnei to poplars

Laboratory studies investigating the host range and pathogenicity of Marssonina castagnei (Desm. + Mont.)Magn. to poplars established that this species was not specifically pathogenic to P. alba but also to cultivars of the Sections Aigeiros, Tacamahaca, and inter sectional hybrids Aigciros × Tacamahaca, Leuce × Aigeiros and Lcucc × Tacamahaca. The potential threat of this pathogen could be reduced by routine screening of new selections for resistance.     

Detection of phytoplasma infections in declining Populus nigra ‘Italica’ trees and molecular differentiation of the aster yellows phytoplasmas identified in various Populus species

A disease of Populus nigra‘Italica’ associated with foliar yellowing, sparse foliage, stunting, dieback, and decline was observed in south-western Germany; a witches’ broom disease of Populus alba that is known in other countries was also detected in Hungary and Germany. The aetiology of the diseases was studied by fluorescence microscopy and polymerase chain reaction (PCR) amplification. Using fluorescence microscopy, phytoplasmas could be detected only in P. alba. However, most diseased trees of P. nigra‘Italica’ tested phytoplasma-positive by PCR. In some of the trees the phytoplasma numbers were so low that nested PCR was required to detect the infection. Very low phytoplasma numbers were also observed in diseased Populus tremula. The identity of phytoplasmas from P. nigra‘Italica’ sampled in Germany and France, P. alba and also P. tremula was examined by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified ribosomal DNA. In all poplars, phytoplasmas of the aster yellows group were detected. However, three different RFLP groups were identified that consisted of (1) French strains from P. nigra‘Italica’, (2) German strains from P. nigra‘Italica’ and (3) strains from P. alba and P. tremula. The profile observed in the last group was probably the result of sequence heterogeneity in the two 16S RNA genes.     

Growth and root sucker ability of field-grown transgenic poplars overexpressing xyloglucanase

Xyloglucan is thought to be a key hemicellulose cross-linking adjacent cellulose microfibrils in plant cell walls. The growth traits of transgenic poplars (Populus alba) with decreased xyloglucan from overexpression of Aspergillus aculeatus xyloglucanase were characterized during a 4-year field trial. The field-trial site consisted of two blocks, a fertile soil block and a non-fertile soil block, determined by soil analysis. In the fertile block, the growth of aboveground biomass of the transgenic poplars was reduced to 24?C44?% compared to that of wild-type poplars, in contrast to the growth seen in chamber and greenhouse conditions. In the non-fertile block, the aboveground biomass of transgenic poplars was also smaller than that of the wild-type poplars. Because poplars reproduce asexually by root suckers, we also compared the formation of root suckers from transgenic and wild-type poplars. Root suckers formed less frequently from transgenic poplars than from wild-type poplars. The growth rates of root suckers from transgenic poplars were also slower than those from wild-type poplars. The results showed that constitutive degradation of xyloglucan impairs poplar growth and vegetative reproduction ability.     

Resistance of improved Pinus monticola and some other white pines to the blister rust fungus,Cronartium ribicola,of Hokkaido,Japan

Resistant progenies of Pinus monticola, improved in the northwestern USA, and Asian and European white pines were inoculated with the blister rust fungus, Cronartium ribicola, of Hokkaido. No resistance was observed in P. monticola but the other pines remained healthy. This shows that the resistance of white pines to the blister rust fungus is limited to a particular pathogenic race and a different race of the rust fungus exists in Hokkaido.     

Poplar growth and wood production on a grassland irrigated for decades with potato starch wastewater

Irrigation of grasslands with potato starch wastewater causes changes in soil quality parameters, often resulting in decreased crop yields and sometimes causing animal diseases. Reduced agricultural income leads to designation of such grasslands for afforestation aimed at production of bioenergy crops and improvement of soil quality. In this study, Populus alba L. and seven different poplar clones were planted in 2009 on the grassland irrigated in 1973–2008 with potato starch wastewater and in non-irrigated, experimental forest. The survival, growth and wood production potential of planted poplars after 2–3 growing seasons were analyzed. Morphophysiological parameters of leaves, nutrients in leaves and fine roots, sugars in fine roots, leaf rust resistance, frost hardiness and their effects on growth were also determined. Stem diameter and stem volume index of poplars growing on the grassland were higher than in the experimental forest. Only Populus deltoides Bartr. × P. maximowiczii Henry clone ‘Eridano’ (in short ERI) had a survival rate of 100 % and the highest values of stem diameter (77 mm) and stem volume index (0.034 m³) in comparison with other poplars in both sites after 3 growing seasons. Clone ERI was also characterized by a high leaf rust resistance and frost hardiness. The results suggest that clone ERI is suitable as a bioenergy crop on grasslands irrigated with potato starch wastewater. Afforestation of the grassland improved the fertility of the soil by increasing concentrations of soil organic matter and availability of mineral nutrients (N, P, K, S and Fe).     

Field ready: Development of a rapid LAMP-based colorimetric assay for the causal agent of white pine blister rust,Cronartium ribicola

The invasive fungal pathogen Cronartium ribicola causes white pine blister rust which is considered one of the most destructive diseases of five-needle (white) pines in North America. The disease has a life cycle that requires two hosts: white pines and Ribes spp., although other non-Ribes species, including Castilleja and Pedicularis, have been demonstrated as alternate hosts as well. Detection of this disease can be difficult because of the ephemeral nature of sporulation on pine hosts with ambiguity in other symptoms, and the alternate hosts for C. ribicola can also be an alternate host for other pine rust species. We used the previously published C. ribicola genome and species-specific real-time PCR assay to develop a field-ready loop-mediated isothermal amplification (LAMP) specific colorimetric assay for this pathogen. Specificity results across regionally identified pine rust pathogens showed the assay is highly specific to C. ribicola and can detect as little as 40 pg of pathogen DNA. We also developed a simple DNA extraction method that works with several tissue types (bark/phloem, aeciospores, and urediniospores/telia) to prepare the DNA samples for the LAMP assay. The DNA extraction and LAMP assay take ~70 min to complete and require a relatively small investment in equipment. This tool enables quick and efficient detection of white pine blister rust.     

Growth response to the stress of low osmotic potential,salinity and high pH in cultured shoot of Chinese poplars

In vitro cultured shoots of three Chinese poplar species (Populus tomentosa, P. alba cv. Pyramidalis andP. euphratica) and an improved poplar (FS-51), as a control, were exposed to the stress of low osmotic potential, salinity and high pH, and the shoot growth was measured.P. tomentosa andP. euphratica could grow even under the high osmotic stress (0.5m mannitol; −1.7 MPa).P. euphratica shoots which were previously cultured on the medium containing 0.7m mannitol (−2.4 MPa) recovered its growth after being transplanted to the medium without mannitol. The shoot growth ofP. euphratica was not so reduced even on the medium containing 100 mM NaCl. However, growth of the other poplars markedly reduced with 10 mM NaCl. Medium pH (pH 5.8–10) also did not affect the shoot growth ofP. euphratica. These poplars have different tolerance to the stresses in cultured shootin vitro. P. euphratica was extremely tolerant to the stresses which are possible to damage plants in dry land. A part of this study was presented at the 104th Annual Meeting of the Japanese Forestry Society. (1993).     

Differences in the rust resistance of greenhouse and outdoor-grown white birch species,Betula spp.

The 2-year-old seedlings of five different white birch species (Betula platyphylla, Betula papyrifera, Betula pubescens, Betula pendula (two types) and Betula resinifera x Betula pendula) grown both in a greenhouse and outdoors, were inoculated in a leaf disc assay with two different birch rust (Mel-ampsoridium betulinum) isolates from B. pendula and B. pubescens. The resistance of these birch species varied significantly. Resistance to the B. pubescens rust isolate was not related to the resistance of the B. pendula rust isolate. The behaviour of a birch genotype grown in the greenhouse did not correspond to the behaviour of the same genotype grown outdoors. The outdoor growth environment greatly increased the contents of soluble proteins, rubisco, chloro-phyll and nitrogen in the leaves of diploid birch species (B. platyphylla, B. pendula and B. resinifera x B. pendula). For tetraploid and pentaploid species (B. pubescens and B. papyrifera, respectively) there was no such clear difference in the leaf physiological status between the seedlings grown outdoors and in the greenhouse. The C:N ratio was higher for the greenhouse-grown seedlings in all the birch species, but the difference was significant only with the diploid species. The incidence of rust in the birch species did not correlate with any of the leaf physiological parameters studied. The adaptability of birch genotypes to the environment in relation to their resistance to birch leaf rust is discussed.     

Spatial patterns and environmental factors affecting the presence of Melampsorella caryophyllacearum infections in an Abies alba forest in NE Spain

The presence of trunk swellings caused by the rust fungus Melampsorella caryophyllacearum was systematically surveyed in an Abies alba forest (Irati, NE Spain), using 1237 circular plots (diameter = 18 m). The relationship between fungal presence and several abiotic (aspect, elevation, distance to the nearest river and slope) and biotic factors (basal area of A. alba and/or Fagus sylvatica, shrub, fern and herb cover) was assessed through correlation and ordination analyses. Additionally, the spatial pattern of the presence of diseased trees was described using Ripley's K function. Southern‐aspect plots had a significantly lower presence of diseased trees than plots‐oriented north, east and west. Plots with diseased trees were located at a significantly lower elevation, and at a shorter distance to the river than plots without infections. Plots with diseased trees had almost twice the average A. alba basal area, and less average F. sylvatica basal area than plots without diseased trees. However, similar mean values of slope and shrub, fern and herb cover were found in both types of plots. The disease showed spatial aggregation in patches with a mean radius of ca. 900 m. The implications of the results for disease management are discussed.     

Analysis of isoperoxidases ofPopulus spp. in Heilongjiang Province

Isoperoxidases of fourteen species of poplars in Heilongjiang Province were analyzed by polyacrylamidegel disc-electrophoresis. The major zymogram of petioles of poplars was consisted of twelve kinds of isoenzymes, in which 35% zymogram was just the same and represented the specialized zymograms of poplars. The results of the experiments showed that the closer the affinity relationships of the species were, the greater the similar values between the zymograms were. There were very great similar values among natural hybrid species. But in sect.Tacamahaca Spach, the similar values betweenPopulus koreana Rehder andP. nakaii Skv. were obviously smaller. Isoenzyme analysis provided new information for plant classification on the molecular level. Responsible editor: Zhu Hong     

Terpene response of Picea abies and Abies alba to infection with Heterobasidion s.l.

The monoterpene composition of Picea abies and Abies alba resin was analysed in relation to growth by Heterobasidion spp. Fifteen‐year‐old P. abies and A. alba trees were inoculated on branches with three species of Heterobasidion annosum s.l. After 4 months of incubation, each host was colonized to a significantly greater degree by the pathogen specific to that host (H. parviporum on P. abies, H. abietinum on A. alba) than by the other fungi. Analysis of the enantiomeric monoterpene profiles in the spruce and fir showed that the response in terms of the relative proportions of the monoterpene compounds in the resin differed between tree species. Following challenge with Heterobasidion spp., A. alba trees did not show changes in monoterpene composition in addition to those in the wounding response (increase in (−)‐α‐pinene and (−)‐camphene, and decrease in β‐phellandrene). In P.abies, (−)‐α‐pinene, (+)‐α‐pinene and δ‐3‐carene increased following Heterobasidion attack but not after wounding alone.     

Research on poplar canker (Xanthomonas populi subsp. populi) in the Netherlands. Part III

A new inoculation method for testing the resistance of poplars to bacterial canker caused by Xanthomonas populi subsp. populi (Ridé) Ridé & Ridé is discussed. The procedure is based on a more natural infection method and involves spraying plants in spring with bacterial suspensions.     

Canker disease of willow and poplar caused by Cryptosphaeria pullmanensis recorded in China

A new canker disease of Salix alba and Populus alba has been observed in Xinjiang, China. Black circular spots on dead branches and stems are the symptoms of the disease. Sixty‐seven isolates recovered from Salix matsudana, S. alba and Populus alba were identified as Cryptosphaeria pullmanensis based on morphological features and multigene phylogeny. Pathogenicity tests were performed on S. alba and P. alba stems using the C. pullmanensis isolates. Cankers on and Cryptosphaeria pullmanensis of C. pullmanensis from the stems fulfilled Koch's postulates and confirmed C. pullmanensis as the causal agent of the canker disease. C. pullmanensis is characterized by its yellow stromatic tissue surrounded by a black conceptacle with regularly arranged multiple locules sharing common walls and hyaline, allantoid, aseptate conidia (mean size 7.42 × 1.72 μm). This is the first report of C. pullmanensis causing Cryptosphaeria canker in China, and S. alba and P. alba are new host records for C. pullmanensis.     

Variation in Melampsoridium resistance among European white-birch clones grown in different fertilization treatments

Resistance of 2-year-old plantlets of seven European white birch, Betula pendula, clones to birch rust, Melampsoridium betulinum, was studied in field experiments and in a leaf-disc bioassay. In addition, rust resistance of plantlets growing in a nursery under three fertilization treatments was tested. The birch clones clearly varied in their levels of resistance. One of the clones was consistently the most resistant, and two were very susceptible. Plantlets growing in the lowest fertility treatment were the most resistant. Clone × fertilization interaction was small. The plantlets grown in the lowest fertilization treatment, in particular, deviated from the other treatments. Generally good rust-resistance correlations were obtained between different experiments. The leaf-disc bioassay was an effective way of determining the field rust resistance of birch clones. The possibility of trade-offs between rust resistance and tree growth is discussed.     

Dothistroma septosporum on firs (Abies spp.) in the northern Baltics

In the spring of 2008, Dothistroma septosporum, a damaging quarantine pathogen of pine foliage, was documented in Järvselja (southeastern Estonia), apparently for the first time, on white fir (Abies concolor). Thus, a new host species and genus for the fungus were recorded. Infection of white fir by D. septosporum, and also of some other fir species, was monitored annually on a transect proceeding from southeastern Estonia to northern Latvia. As a result, D. septosporum was detected also on silver fir (Abies alba). It is not clear why this fungus with a worldwide distribution appeared on this new host genus definitely in northern Europe.     

Pedicularis and Castilleja are natural hosts of Cronartium ribicola in North America: a first report

White pine blister rust disease, caused by the introduced pathogen Cronartium ribicola, has severely disrupted five‐needled pine ecosystems in North America. A 100‐year effort to manage this disease was predicated in part on the premise that the pathogen utilizes only species of Ribes (Grossulariaceae) as alternate hosts on this continent. The current study presents the first conclusive demonstration that some species in the family Orobanchaceae (Pedicularis racemosa and Castilleja miniata) are functioning as alternate hosts in a natural ecosystem of North America. This finding has implications for improving our understanding of epidemiology, pathogen adaptation and host–pathogen interactions within white pine blister rust.