Seedling response of two trembling aspen (Populus tremuloides) families to infection by Hypoxylon mammatum

Open-pollinated progenies from two trembling aspen (Populus tremuloides) families with a 30-year field history of 91% (putatively susceptible, PS) and 45% (putatively resistant, PR) mortality, respectively, caused by Hypoxylon mammatum, were grown in a greenhouse for 3 and 9 months. Seedlings were inoculated with two isolates of H. mammatum, representing high and low aggressiveness, through a stem wound. Beginning 4 days after inoculation, host responses were examined on 312 seedlings per family for a period of 5 weeks on 3- and 9-month-old seedlings. Significant differences in host responses were observed between the two isolates, with the more aggressive isolate allowing less callus formation, causing more tissue necrosis and producing longer cankers than the less aggressive isolate on both aspen families. Between-family difference was much greater than within-family difference for tissue necrosis, callus formation and canker length for both seedling ages. When the 3-month-old seedlings were challenged with H. mammatum, the PR family showed greater tissue necrosis, longer cankers, and less callus formation than the PS family. However, when the more lignifìed 9-month-old seedlings of the same families were inoculated, the PR seedlings had much less tissue necrosis, shorter cankers, more callus formation and less seedling mortality than the PS family. The results indicate that artificial inoculation of seedlings in a greenhouse can differentiate two aspen families at the seedling stage, but their correlation to field performance is highly dependent upon the seedling age. Host responses of older seedlings may be a better indicator of field performance.     

Responses of Sitka spruce of different genetic origins to inoculation with Heterobasidion annosum: lesion lengths,fungal growth and development of the lignosuberized boundary zone

Three genetically distinct groups of Sitka spruce, open‐pollinated Queen Charlotte Island provenance, A13 selected seed and M0044 half‐sib mixture, were wounded alone or wounded and inoculated with Heterobasidion annosum sensu stricto on the lower stems. Growth of the pathogen and lesion formation was compared in the three genetic groups after treatment. No differences in the rate of colonization of the three genetic groups were observed over a 40 day period; lesion lengths in the bark and sapwood correlated closely. Moreover, lesions were considerably longer in inoculated plants than in those which were wounded only. No correlations were found within or between host genetic groups in the numbers or total areas of resin canals present in the first 18 mm from the wound in bark tissues for the three host genetic groups. Formation of the ligno‐suberized boundary zone (LSZ), however, was inhibited in the bark of inoculated plants, being first detected later and at a greater distance from the wound/inoculation point in the presence of H. annosum than in plants that were wounded only. Thickness of the suberin cell layers within the LSZ of M0044 plants was greater in wounded and inoculated, than in wounded only plants. The significance of these findings in relation to detecting spruce genotypes potentially resistant to H. annosum is discussed.     

Effects of browsing on recruitment and mortality of European aspen (Populus tremula L.)

Abstract

High browsing pressure may reduce or halt regeneration of browsing-prone tree species. In this study, exclosures were used to quantify the effects of browsing on vital demographic parameters in European aspen (Populus tremula L.) stands. The experiment focused on stands located in managed forest landscape with densities of 0.85 moose (Alces alces L.)km^?2. Other herbivores such as mountain hare (Lepus timidus L.) were present at densities common for this region and roe deer (Capreolus capreolus L.) at low densities. Recruitment and mortality of aspen ramets were recorded before and after the growing season, and over four consecutive years. Seven to 19% of the ramets within browsing reach were browsed annually. No differences were found in recruitment or mortality attributable to browsing per se. Instead, there were large among-stand variability in vital population parameters unrelated to browsing incidence. In general, aspen stands with high ramet density showed higher recruitment rates than stands with low ramet density. There was substantial variability in self-replacement capability unrelated to ecological factors. The results suggest that more attention should be given to finding and managing stands with high capacity for suckering, as these stands may have a disproportionate impact on aspen recruitment at the landscape scale.     

Foliar susceptibility of eastern oak species to Phytophthora infection

Seven different Phytophthora species were used to test the foliar susceptibility of the common eastern US oak species and understory plants to Phytophthora infection. The Phytophthora species employed were Phytophthora cambivora, Phytophthora cinnamomi, Phytophthora citricola, Phytophthora europaea, Phytophthora quercetorum, Phytophthora quercina‐like and Phytophthora sp1. Inoculation of detached‐leaves with agar plugs containing mycelia of Phytophthora provided an estimate of their relative susceptibility. Lesions were always greater when foliage was wounded and young. On deciduous plants, lesion sizes were considerably reduced with the increasing foliar age, although with evergreen plants lesion sizes remained similar regardless of foliar age when more aggressive isolates were tested. Infections seldom resulted when foliage was not wounded. With young and mature foliage, P. citricola usually produced the largest lesions. Young foliage of Quercus rubra was the most susceptible to infection followed by Castanea dentata for both wounded and non‐wounded inoculations. Mature foliage of Hamamelis virginiana, Kalmia latifolia and Quercus alba were the most susceptible to wound and non‐wound inoculations.     

Wound response of Eucalyptus clones after inoculation with Cryphonectria cubensis

Twenty-five different Eucalyptus grandis clones were artificially wounded and inoculated with a virulent isolate of Cryphonectria cubensis. The capacity of wounds to close through callus production was correlated with the relative susceptibility of these clones to infection by C. cubensis. Clones with the greatest capacity to close wounds were those that were also most tolerant to C. cubensis infection. Those with a lower capacity to close wounds were most susceptible to Cryphonectria canker.     

Effects of Ophiostoma ulmi and Ophiostoma novo-ulmi culture filtrates on elm cultures from genotypes with different susceptibility to Dutch elm disease

Cell cultures of callus tissue cultures obtained from four elm genotypes (Ulmus minor; Ulmus minor×Ulmus pumila; [Ulmus carpinifolia×Ulmus glabra] × [Ulmus wallichiana×Ulmus glabra]; and Ulmus pumila), either susceptible or resistant to Dutch elm disease (DED) were exposed to culture filtrates of Ophiostoma ulmi and Ophiostoma novo-ulmi. Elm cells were largely affected by crude culture filtrate incorporated into the media. However, the correlation between ‘in vivo’ cell resistance and growth in the presence of culture filtrate was poor: the effects of fungal media components were greater than that exerted by fungal exotoxins. Therefore, it is concluded that these ‘in vitro’ assays cannot be used to evaluate resistance sources to DED in elms, or to assess specific pathogenicity of fungal isolates.     

Responses of Cupressus and Chamaecyparis callus tissues to inoculations with Seiridium cardinale

Inoculations of callus generated from micropropagated shoots of Cupressus sempervirens, Cupressus macrocarpa, Cupressus torulosa and Chamaecyparis lawsoniana showed that callus of species resistant or relatively resistant to Seiridium cardinale supported significantly less surface growth of the pathogen than callus of susceptible species. In dual cultures of Cupressus sempervirens or Cupressus torulosa callus with S. cardinale, inhibition of fungal growth towards the callus was correlated with known field resistance of these host species. Lignification, accumulation of polyphenolics and possible suberization of cell walls were detected in inoculated callus. This response was also correlated with known field resistance of the tested host species.     

Plant regeneration in <Emphasis Type="Italic">Actinidia polygama</Emphasis> Miq. by leaf,stem, and petiole culture with zeatin,and from stem-derived calli on low-sucrose medium

We examined the effect of zeatin on the formation of shoot buds from explants and callus tissues derived from stem segments of Actinidia polygama Miq. (matatabi or silver vine). Stem and petiole segments cultured on combined broad-leaved tree medium and woody plant medium (BW medium) supplemented with zeatin for 40 days formed many shoot buds. Callus tissues derived from the stem segments and subcultured on BW medium supplemented with 6-benzyladenine and 1-naphthaleneacetic acid formed shoot buds when the medium contained 13.7µM zeatin. BW medium containing low concentrations of sucrose strongly induced the formation of shoot buds from the callus.     

Propagation of trembling aspen and hybrid poplar for agroforestry: potential benefits of elevated CO2 in the greenhouse

We examined the usefulness of elevated CO₂ in the greenhouse to aid in early selection of genotypes and in the propagation of Populus tremuloides Michaux (aspen) and hybrid poplars for agroforestry, afforestation, or reclamation. Growth in elevated (800 ppm) vs ambient (375 ppm) CO₂ for 95 days resulted in greater height (14%), stem caliper (16%), overall biomass, and proportional allocation of biomass to roots as well as elevated net assimilation and water-use-efficiency. Aspen clones selected for superior growth (based on phenotypic selection) broke bud significantly earlier than unselected clones under both CO₂ levels; superior clones were also taller with greater stem caliper under both CO₂ treatments (but not significant). Under ambient CO₂ male aspen were taller than females while under elevated CO₂ female aspen were taller than males and also had greater caliper (but not significant). Hybrid poplar grown under elevated (vs ambient) CO₂ broke bud significantly earlier and had significantly greater net assimilation and water-use efficiency; they were also taller with greater caliper (but not significant). Differences in performance among the eight hybrid clones were enhanced by growth under elevated CO₂. Under ambient CO₂, P38P38 was the clone that broke bud earliest, was tallest with greater caliper and had a lower shoot:root biomass ratio. Sargentii, on the other hand, was the shortest with the smallest caliper. The implications of these results need to be considered in the context of using aspen and hybrid poplar for large-scale agroforestry, afforestation and reclamation across Canada. This revised version was published online in August 2006 with corrections to the Cover Date.     

Relationship between canker size and wood starch in American chestnut

Chestnut trees (Castanca dentata) were wounded in May and inoculated with Cryphonectria parasitica. In August, canker area was measured and the surfaces of longitudinal dissections of stem wood were stained with I₂KI to reveal the presence of starch. Sections with the most starch had smaller cankers than sections with no starch. Results show that starch reserves were related to canker size and therefore may play a role in the tree's ability to respond to infection.     

Formation of the ligno-suberized barrier zone and wound periderm in four species of European broad-leaved trees

The development of wound periderm after mechanical wounding was examined in the barks of Acer pseudoplatanus, Betula pendula, Fagus sylvatica and Sorbus aucuparia. Following the initial necrosis of cells immediately surrounding the wounds, development of the ligno-suberized barrier zone was detected within 7 days. Suberin deposition was observed in this tissue in A. pseudoplatanus, S. aucuparia and B. pendula 7 days after wounding, but not in F. sylvatica until day 14. By 28 days after wounding, extensive thickening of the walls of these cells had occurred. Differentiation of the wound periderm was detected immediately abutting the ligno-suberized barrier-zone tissues in all species, with extensive development by 28 days. Fifty-six days after wounding, the wound periderm was fully formed and completely confluent with the original periderm. Development of the wound response was similar in all species, with differences only in the timing of first detection of suberin.     

Migration of Bursaphelenchus xylophilus in cortical and xylem axial resin canals of resistant pines

Migration of the pine wood nematode (PWN), Bursaphelenchus xylophilus, in susceptible and resistant pines was investigated at the tissue level. PWN was inoculated onto the top cross‐cut surface of 20‐cm stem cuttings of susceptible Pinus thunbergii and resistant pines (P. strobus, P. rigida and P. thunbergii of a resistant family Namikata‐(t)‐73 (half‐sib)). PWNs were mainly distributed in cortical resin canals of susceptible P. thunbergii down to 15 cm from the inoculated surface by 6 h after inoculation (HAI) and all tissues (including cortical and xylem resin canals) down to the bottom at 192 HAI. In P. strobus, P. rigida and P. thunbergii family Namikata‐(t)‐73 (half‐sib), PWN was distributed in cortical resin canals down to 5 cm by 6 HAI and down to the base at 192 HAI. However, the distribution of PWN in xylem resin canals of the resistant pines was restricted near inoculated surfaces down to 5 cm, even at 192 HAI. These results demonstrated that migration of PWN in resistant pines was slowed in cortical resin canals and restricted in xylem axial resin canals, features which may be associated with the resistance.     

Inhibiting factors of symptom development in several Japanese red pine (<Emphasis Type="Italic">Pinus densiflora</Emphasis>) families selected as resistant to pine wilt

Pinus densiflora and P. thunbergii, native to Japan, are highly susceptible to pine wilt disease caused by infection with a pine wood nematode (Bursaphelenchus xylophilus). Trees of these susceptible species have occasionally been found surviving in forests that are extensively damaged by this disease. Seedlings from a part of surviving trees that were selected as resistant families indicate lower mortality rates after the infection. The factors that prevent the symptoms from developing in resistant families of a susceptible species, P. densiflora, as based on the analysis of the pathogens behavior in the tree tissue and the anatomy of the resistant families, are presented in this paper. Nematode populations remained lower in the stems of seedlings from resistant families of P. densiflora than in the stems of non-resistant families. Areas dysfunctional in water transport developed in the stems of resistant families, but did not reach a size large enough to seriously block the ascent of sap. These results suggest that there are systems within the seedlings that prevent nematode migration and reproduction. The 2-year-old seedlings from resistant families of P. densiflora, however, did not suppress the pathogen activity. Numerous branches are a visible characteristic in the seedlings of some resistant families. The arrangement of the resin canals, the only channels in the tree to the pathogen migration, was disoriented at the joints between the branches and the main stem. Such a structure may be effective as a barrier to nematode dispersal.     

Rooting of Hybrid Clones of Populus tremula L. × P. tremuloides Michx. by Stem Cuttings Derived from Micropropagated Plants

Propagation costs could be cut by replacing part of the micropropagation process with steps involving more traditional techniques. This study explored possibilities for improving existing vegetative propagation techniques for aspen using stem cuttings obtained from micropropagated plants. Vegetative propagation through stem cuttings was studied in 10 micropropagated hybrid aspen clones (Populus tremula L.     

Comparison of the modified Weibull and Richards growth function for developing site index equations

A site index model based on the Weibull cumulative distribution function was fitted to three stem analysis data sets of black spruce (Picea mariana (Mill) BSP), white spruce (Picea glauca (Moench) Voss) and aspen (Populus tremuloides Michx.). The resulting equations were compared with those based on the modified Richards' (1959) biological growth function (Ek 1971; Payandeh 1977, 1978). The Weibull-type model performed as well as or better than the biological growth function for white spruce and aspen, respectively, but it performed less accurately in the case of black spruce. The Weibull-type model appears to behave better, i.e., it produces more realistic height growth patterns, than the Richards function upon extrapolation particularly at young ages.     

Fourier transform-infrared spectroscopy as a new method for evaluating host resistance in the Dutch elm disease complex

Resistance of elms (Ulmus spp.) to the pathogenic fungus Ophiostoma novo-ulmi Brasier depends on chemical and anatomical factors that confine the spread of the pathogen in the vascular system of the host. This study focused on detecting chemical differences in 4-year-old Ulmus minor Mill. seedlings before and after inoculation with a virulent O. novo-ulmi isolate. According to symptom development over 60 days, the trees were divided into resistant (0-33% wilting) and susceptible (67-100% wilting) groups. Histochemical tests and Fourier transform-infrared (FT-IR) spectroscopy analysis were performed on transverse sections of 2-year-old twigs, 2 days before and 40 days after inoculation. Although histochemical tests did not clearly discriminate susceptible from resistant elms, chemical differences between resistant, susceptible and control trees were detected by FT-IR. The average spectrum for resistant tree samples had higher absorbance peaks than the spectra from the susceptible and control samples, indicating increased formation of lignin and suberin. The roles of lignin and suberin in the resistance of the elms against O. novo-ulmi and the usefulness and sensitivity of the FT-IR technique for analyzing metabolic changes caused by pathogens in plants are discussed.     

Does mechanical site preparation affect trembling aspen density and growth 9–12 years after treatment?

Trembling aspen (Populus tremuloides Michx.) density and growth were assessed 9–12 years after stand establishment to determine whether mechanical site preparation (MSP) affects crop tree quality. Study sites were either treated with disc trenching or ripper ploughing and planted with white spruce (Picea glauca (Moench) Voss) seedlings immediately after harvest (treated) or were undisturbed since harvest (control). Stands were surveyed during the summer of 2002 with standard regeneration survey plots. Results show that aspen stem density and height were lower in MSP-treated areas relative to untreated areas. Diameter growth rates were unaffected by treatment, however the percentage of stem discolouration was higher in untreated control stands compared to site prepared areas. The results of this study indicate that there are no long-term benefits to carry out MSP for aspen promotion. However, as MSP does not appear to seriously harm the aspen crop, we suggest that this treatment can still be used on sites where aspen densities may be low without treatment (e.g., sites with extremely low soil temperatures, poor soil aeration, or vigorous competitive vegetation) or where a mixture of aspen and planted spruce are desired.     

Physiological and biochemical differences among Ulmus minor genotypes showing a gradient of resistance to Dutch elm disease

Dutch elm disease (DED) spread across Europe and North America in the 20th century killing most natural elm populations. Today, breeding programmes aim at identifying, propagating and studying elm clones resistant to DED. Here, we have compared the physiology and biochemistry of six genotypes of Ulmus minor of variable DED resistance. Leaf gas exchange, water potential, stem hydraulic conductivity and biochemical status were studied in 5‐year‐old trees of AB‐AM2.4, M‐DV2.3, M‐DV2 × M‐CC1.5 and M‐DV1 and 6‐year‐old trees of VA‐AP38 and BU‐FL7 before and after inoculation with Ophiostoma novo‐ulmi. Leaf water potential and net photosynthesis rates declined, while the percentage loss of hydraulic conductivity (PLC) increased after the inoculation in susceptible trees. By the 21st day, leaf predawn and midday water potential, stomatal conductance to water vapour and net photosynthesis rates were lower, and PLC was higher in trees of susceptible (S) genotypes inoculated with the pathogen than in control trees inoculated with water, whereas no significant treatment effect was observed on these variables in the resistant (R) genotypes. Fourier transform infrared spectroscopy analyses revealed a different biochemical profile for branches of R and S clones. R clones showed higher absorption peaks that could be assigned to phenolic compounds, saturated hydrocarbons, cellulose and hemicellulose than S clones. The differences were more marked at the end of the experiment than at the beginning, suggesting that R and S clones responded differently to the inevitable wounding from inoculation and repeated sampling over the experimental course. We hypothesize that a weak activation of the defence system in response to experimental wounding can contribute to the susceptibility of some genotypes to O. novo‐ulmi. In turn, the decline in shoot hydraulic conductivity and leaf carbon uptake caused by the infection further exacerbates tree susceptibility to the fungus.     

Preliminary studies on establishment of genetic transformation system in loblolly pine

A transformation system was established for loblolly pine (Pinus taeda L.) mature zygotic embryos usingAgrobacterium tumefaciens. The gene coding for the β-glucuronidase (GUS) gene was introduced into loblolly pine tissues and its transient expression was detected with histochemical staining. The influences of different genotypes.Agrobacterum concentrations. and cocultivation time on GUS expression and kanamycin resistant callus and shoot regeneration were investigated. The results showed that the highest GUS expression frequency (16.3%) and shoot regeneration frequency (78%) were obtained from genotype 9–1003 withAgrobacterium concentration decreased 9 times and cocultivation time of 56 hours respectively. GUS expression was obtained in all genotypes tested. The successful expression of the GUS gene in different genotypes suggested that it will be a useful transformation system for loblolly pine. (Responsible Editor: Chai Ruihai)