毛白杨形成层的活动周期及其POD同工酶的变化

北京地区生长的毛白杨 (PopulustomentosaCarr .)在春季芽膨大前 ,形成层带已经进入了恢复活动期。在芽展开后一周形成层进入活动期 ,细胞开始分裂 ,细胞层数也有所增加 ,同时分化出未成熟的韧皮部细胞。未成熟韧皮部细胞的数量在形成层活动初期快速增长 ,随后是一个持续而缓慢的连续过程。木质部的产生比韧皮部要晚 3周 ,未成熟的木质部细胞层数在开始变化不大 ,5月底～ 6月底 ,增长速度明显高于初期 ,随后便急剧下降。成熟木质部细胞的出现比成熟的韧皮部细胞要晚约一个月。与其他研究结果不同 ,毛白杨在4月底到 6月底这段时间 ,形成层细胞层数迅速下降 ,未成熟韧皮部细胞也略有减少 ,而未成熟和成熟的木质部细胞则大量增加。木质部在 9月底停止分化 ,较韧皮部要晚约 2 0d。 9月底形成层进入休眠期后 ,形成层细胞层数基本保持不变 ,直到翌年春天恢复活动。毛白杨形成层区域的过氧化物酶 (POD)同工酶在形成层活动期其活性低、酶带少 ,而恢复活动期和休眠期其活性高且酶带多 ,组织化学反应还表明 ,不同时期POD在不同的细胞和组织中的活性不同。本文对POD同工酶在形成层活动周期中的变化与其组织分化的关系进行了讨论。     

Effect of nitrogen on the seasonal course of growth and maintenance respiration in stems of Norway spruce trees

To determine effects of stem nitrogen concentration ([N]) on the seasonal course of respiration, rates of stem respiration of ten control and ten irrigated-fertilized (IL), 30-year-old Norway spruce trees (Picea abies (L.) Karst.), growing in northern Sweden, were measured on seven occasions from June 1993 to April 1994. To explore sources of seasonal variation and mechanisms of fertilization effects on respiration, we separated total respiration into growth and maintenance respiration for both xylem and phloem bark. Stem respiration increased in response to the IL treatment and was positively correlated with growth rate, volume of living cells and stem nitrogen content. However, no significant effect of IL treatment or [N] in the living cells was found for respiration per unit volume of live cells. Total stem respiration during the growing season (June to September) was estimated to be 16.7 and 29.7 mol CO(2) m(-2) for control and IL-treated trees, respectively. Respiration during the growing season accounted for approximately 64% of total annual respiration. Depending on the method, estimated growth respiration varied between 40 and 60% of total respiration during the growing season. Between 75 and 80% of the live cell volume in the stems was in the phloem, and phloem maintenance accounted for about 70% of maintenance respiration. Because most of the living cells were found in the phloem, and the living xylem cells were concentrated in the outer growth rings, we concluded that the best base for expressing rates of stem growth and maintenance respiration in young Norway spruce trees is stem surface area.     

Methyl jasmonate and oxalic acid treatment of Norway spruce: anatomically based defense responses and increased resistance against fungal infection

To study the effect of chemical pretreatment on conifer resistance, 13-year-old Norway spruce (Picea abies (L.) Karst.) trees were treated with methyl jasmonate (MJ) or oxalic acid (OxA) on the outer bark and inoculated with the pathogenic blue-stain fungus Ceratocystis polonica (Siem.) C. Moreau 4 weeks later. Both chemicals significantly reduced symptoms of fungal infection, but MJ was more effective than OxA (51 versus 18% reduction in length of necrotic lesions in the phloem relative to untreated control trees). Anatomical examination of treated stem tissues showed that MJ induced extensive formation of traumatic resin ducts in the xylem and extra polyphenolic parenchyma (PP) cells in the secondary phloem between the cambium and the regular annual PP cell layer. No traumatic resin ducts were formed after treatment with OxA, and the coverage of extra PP cells in OxA-treated tissues was not significantly higher than in the controls. The anatomically based defense reactions induced by MJ were similar to the reactions observed after pathogen infection, mechanical wounding and bark beetle attack. Neither MJ nor OxA had apparent phytotoxic effects on Norway spruce at the concentrations used, with needle and stem tissues of all trees appearing normal without visible symptoms of toxicity. However, trees treated with MJ had 30% less radial sapwood growth than control trees. In conclusion, MJ treatment of Norway spruce appears to have practical potential as a tool for increasing plant resistance to fungal infection, but with a modest reduction in sapwood growth.     

Initial reactions in sapwood of Norway spruce and Scots pine after wounding and infection by Heterobasidion parviporum and H. annosum

The xylem surface of seedlings, stem material and roots of Norway spruce (Picea abies) and Scots pine (Pinus sylvestris) were inoculated with strains of Heterobasidion annosum s. str. and H. parviporum s. str. The depth of necrosis in wounded spruce increased at a linear rate for at least seven weeks of incubation, but the rate of necrotic spread was significantly faster in infected wounds. In wounded pine the necrosis was maintained at a more superficial level for several weeks. Both spruce and pine sapwood were initially infected by hyphae of both species. In spruce, the hyphae advanced at a constant rate behind the necrotic front. On the contrary, after 1–2 weeks living H. parviporum hyphae were rare in pine rays. Heterobasidion annosum hyphae survived in pine rays, phloem and tracheids, despite a heavy accumulation of phenolics and resins and were able to penetrate into the sapwood at a linear rate although slower than infections in spruce. Histochemistry and quantitative estimates demonstrated that peroxidase activity was initially higher in spruce sapwood than in pine. Within three days of incubation, the activity in spruce sapwood disappeared concurrently with deepening necrosis. However, in pine, in both control and infected samples, there was a significant increase in peroxidase activity in the area surrounding the superficial necrosis, up to the wound surface and in the cambium and phloem around the wound. After wounding and infection, the content of soluble protein increased significantly in wood of older trees but not in seedlings. Infection resulted in an increased formation of lipophilic extractives in both spruce and pine but to a significantly greater degree in the latter, whereas the amount of hydrophilic compounds decreased in both. High‐performance liquid chromatography (HPLC) analyses of lipophilic extracts showed that inoculation of pine with the two species of Heterobasidion increased the amounts of pinosylvin, its monomethylether and several other phenolics as also resinous compounds. The results obtained may be relevant in explaining the known higher resistance of Scots pine to H. parviporum.     

Thermochemical behavior of Norway spruce (Picea abies) at 180–225 °C

Norway spruce (Picea abies) was heated for 2–8 h in the temperature range 180–225 °C, under a steam atmosphere. The chemical analyses of the treated feedstock samples indicated that during heating (total mass loss 1.5–12.5% of the initial DS) carbohydrates (hemicelluloses and cellulose) were clearly more amenable to various degradation reactions than lignin. In addition, major water-soluble products released from the feedstock material during the treatments were classified into several compound groups and changes in the relative mass portion of these groups were monitored by GC during a separate experiment. Received 20 December 1998     

Effects of seed water content and storage temperature on the germination parameters of white spruce, black spruce and lodgepole pine seed

The effect of seed water content (WC) (2–3, 5–6 and 22–25%, on a fresh weight basis), storage temperature (+4, −20, −80 and −196°C) and storage duration (6, 12, 24, 48 and 60 months) on the germination of white spruce (Picea glauca (Moench) Voss), black spruce (Picea mariana (Mill.) B.S.P.) and lodgepole pine (Pinus contorta Dougl. ex Loud. var. latifolia Engelm.) seed was investigated. Germination of white spruce control (untreated) seeds and seeds adjusted to 2–3% and 5–6% WC declined after 48 months of storage at −80 and −196°C, with a further decline at 60 months at −20, −80, −196°C. Germination remained high when control white spruce seeds and seeds with 2–3, 5–6% WC were stored at +4°C, over all storage durations. Generally, black spruce and lodgepole pine exhibited high germination at all storage temperatures at 2–3% and 5–6% WC as well as the control (untreated) seed, for up to 60 months in storage. Germination declined for all three species when seed was conditioned to 22–25% WC. This loss in germination was partially recovered in white spruce seed stored at +4, −20 and −80°C after storage durations of 24, 12 and 48 months, respectively, and in black spruce seeds stored at −20 and −196°C after storage durations of 24 months. Mean germination time (MGT) was relatively constant for all species, under all conditions, except for seed conditioned to 22–25% WC, where MGT increased for white spruce seed stored 48 months at −80 and −196°C, and for black spruce seed stored 24 months at +4 and −80°C and 60 months at −196°C. These results show that the optimal storage temperatures are 4°C for white spruce, and 4, −20, −80, and −196°C for black spruce and lodgepole pine, and 2–6% water content is optimal for all 3 species at these temperatures.     

Control of longitudinal and cambial growth by gibberellins and indole-3-acetic acid in current-year shoots of Pinus sylvestris

We investigated the involvement of gibberellins (GAs) and indole-3-acetic acid (IAA) in the control of longitudinal and cambial growth in current-year shoots of Pinus sylvestris L. Elongating terminal shoots, located at the apex of previous-year (1-year-old) branches in the uppermost whorl on the main stem, were variously decapitated (apical 5 to 10 mm removed), defoliated (all developing needle fascicles removed) and treated with endogenous GA(4/7) or IAA, or both. Shoot length and the radial widths of xylem and phloem were measured, and the concentrations of GA(1), GA(3), GA(4), GA(9) and IAA in the stem were determined by combined gas chromatography-mass spectrometry with deuterated GAs and [(13)C(6)]-IAA as internal standards. Decapitation decreased the production of xylem and phloem and the IAA concentration, but did not alter either longitudinal growth or the concentrations of GAs. Defoliation markedly inhibited shoot elongation, as well as cambial growth, and reduced the concentrations of GA(1), GA(3), GA(4), GA(9) and IAA. Application of GA(4/7) to defoliated shoots promoted longitudinal growth and phloem production, without affecting xylem production or IAA concentration. Application of GA(4/7) and IAA together to decapitated + defoliated shoots increased shoot elongation, xylem and phloem production and IAA concentration, whereas applying either substance alone had a smaller effect or none at all. We conclude that, for elongating current-year shoots of Pinus sylvestris, (1) both the shoot apex and the developing needle fascicles are major sources of the IAA present in the stem, whereas stem GAs originate primarily in the needle fascicles, (2) GAs and IAA are required for both shoot elongation and cambial growth, and (3) GAs act directly in the control of shoot growth, rather than indirectly through affecting the IAA concentration.     

A study on Xingkai Lake pine gall rust

The Xingkai Lake pine (Pinus takahasii Nakai) gall rust caused byCronartium quercuum(Berk.) Miyabc: Shirai is a serious stem rust in the northeast region of China. The alternate host is oak (Quercus mongolica Fisch). Germination of both acciospores and urediospores was optimal at 12°C and occurred over a range of temperatures, from 4°C to 32°C. Teliospores germinated optimally at 16–18°C and over a range of 8–28°C. All spores germinated best under natural light and dark conditions. Direct light inhibited germination even when followed by darkness. The incubation periods of urediospores and teliospores in inoculation experiments were 6–16 days and 3–33 days respectively. Anatomical studies ofCronartium quercuum galls showed that rust hyphae freely ramify through the intercellular spaces within parenchymatous tissues of the cortex, phloem, cambium and xylem. Hyphae are perennial; large haustoria arc cylindrical with rounded or blunt ends. Among the 12 fungicides tested for control of the rust, pine-tar, pine-tar: diesel oil (1:1, 1:3, 1:5), 75% Bravo emulsion and 70% Mancozeb emulsion applied to the galls produced satisfactory results.     

Can the structure of dormant cambium and the widths of phloem and xylem increments be used as indicators for tree vitality?

We investigated the structure and width of the dormant cambium and of the increments of phloem and xylem of Quercus robur to estimate their potential as indicators for tree vitality. The samples were taken from three woodlands, two in Slovenia [Krakovo forest (KRA) and Murska Suma (MUS)] and one in Croatia [Kobiljak (KOB)], with reported tree decline. The number of dormant cells seems to reflect the initial capacity of the cambium to accomplish cell division. With the exception of two trees at KRA, cell production was always higher on the xylem side than on the phloem side. The annual phloem increments were narrower, less variable among trees and with clear lower and upper limits. With increased cambial cell productivity, the share of the xylem in the total annual radial increment increased following a curvilinear function. In trees with an annual radial increment >3.5 mm, the xylem size represented more than 90 % of the total radial growth. The anatomical variables analyzed show that the most limiting environmental conditions seem to prevail at KRA, whereas the conditions at MUS seem to be most favorable in terms of radial growth. Analysis of the width and structure of xylem and phloem increments, the number of dormant cambial cells and their inter-relationships can provide additional information on the vitality of oaks.     

Water content and bark thickness of Norway spruce (Picea abies) stems: phloem water capacitance and xylem sap flow

To determine the relationship between phloem transport and changes in phloem water content, we measured temporal and spatial variations in water content and sucrose, glucose and fructose concentrations in phloem samples and phloem exudates of 70- and 30-year-old Norway spruce trees (Picea abies (L.) Karst.). Large temporal and spatial variations in phloem water content (1.4-2.6 mg mg(dw)(-1)) and phloem total sugar concentration (31-70 mg g(dw)(-1)) paralleled each other (r(2) = 0.83, P < 0.0001 for the temporal profile and r(2) = 0.96, P < 0.008 for the spatial profile), indicating that phloem water content depends on the total amount of sugar to be transferred. Changes in phloem water content were unrelated to changes in bark thickness. Maximum changes in phloem water content calculated from dendrometer readings were only 8-11% of the maximum measured changes in phloem water content, indicating that reversible changes in bark thickness did not reflect changes in internal water relations. We also studied the relationship between xylem sap velocity and changes in bark thickness in 70-year-old trees during summer 1999 and winter 1999-2000. Sap flow occurred sporadically throughout the winter, but there was no relationship between bark shrinkage or swelling and sap velocity. In winter, mean daily xylem sap velocity was significantly correlated with mean daily vapor pressure deficit and air temperature (P < 0.0001, in both cases). Changes in bark thickness corresponded with both short- and long-term changes in relative humidity, in both winter and summer. Under controlled conditions at > 0 degrees C, changes in relative humidity alone caused changes in thickness of boiled bark samples. Because living bark of Norway spruce trees contains large areas with crushed and dead sieve cell zones-up to 24% of the bark is air-filled space-we suggest that this space can compensate for volume changes in living phloem cells independently of total tissue water content. We conclude that changes in bark thickness are not indicative of changes in either phloem water capacitance or xylem sap flow.     

Locating the initial in the vascular cambium of Pinus strobus L. by electron microscopy

Summary The results of this investigation by electron microscopy on the vascular cambium in Pinus strobus L. confirm the results of earlier investigations by light microscopy. First, they showed that differences in the thickness of tangential walls of cambial cells exist and that these differences permit grouping of cells according to the sequence of the previous divisions of the initial. This, in turn, permits the site of the initial cell in the cambial zone to be deducted. The thicker distal tangential wall of the initial in the successive series of tissue production allows for precluding the direction in which the initial function proceeds, toward the xylem or toward the phloem. Second, it was demonstrated that immature xylem cells exist in groups of four, that immature phloem cells exist in pairs, and that at all times more cells are produced toward the xylem than toward the phloem. Third, it was shown that the extra-thick tangential wall in immature xylem serves as a landmark to signal the changeover in the initial function from phloem to xylem production.     

Effect of grain angle on shear strength of glued end grain to flat grain joints of defect-free softwood timber

To evaluate the effect of grain orientation on the adhesive bond strength, three-layered Norway spruce wood specimens were tested in shear. The two axial-oriented outer layers were jointed with the middle layer using three typical glues for load bearing constructions, i.e. one-component polyurethane (PUR), melamine–urea–formaldehyde (MUF) and phenol–resorcinol–formaldehyde (PRF). The grain orientation of the middle layers was varied from 0° (parallel to grain of the surface layer) to 90° (perpendicular to grain of the surface layer) in incremental steps of 10°. Samples with middle layers oriented parallel to the outer layers showed shear values in the range of solid spruce wood. Decreased shear strength values were expected for increased grain angles of the middle layer. However, no explicit tendency was observed for the shear strength in dependence of the grain direction. In general, MUF-bonded samples showed slightly higher shear strength values compared with PUR- and PRF-bonded specimens.     

Pressurized hot water extraction of Norway spruce hemicelluloses using a flow-through system

Norway spruce saw meal was extracted with pressurized hot water at 120–240°C using a flow-through system. Only small amounts of hemicelluloses were extracted at 120–160°C, but dissolution was significantly enhanced when higher extraction temperatures were applied. All hemicelluloses but only 15% of lignin were removed from wood at 220°C, and even less lignin was extracted at lower temperatures. Partial degradation of cellulose seemed to take place only at 240°C. Of the total amount of extracted hemicelluloses, 4–22% was hydrolyzed to monosaccharides. Although the average molar masses of extracted hemicelluloses decreased with increasing extraction temperature, even at 240°C the extracted carbohydrates occurred, on average, as polysaccharides. Polysaccharides with an average molar mass of 31 kDa were obtained at 170°C. The molecular-mass characteristics and yield of carbohydrates depend on the extraction temperature, which should be chosen based on the end use of the isolated hemicelluloses.     

Xylogenesis in black spruce: does soil temperature matter?

In boreal ecosystems, an increase in soil temperature can stimulate plant growth. However, cambium phenology in trees was better explained by air than soil temperature, which suggested that soil temperature is not the main limiting factor affecting xylogenesis. Since soil temperature and snowmelt are correlated to air temperature, the question whether soil temperature directly limits xylogenesis in the stem will remain unresolved without experiments disentangling air and soil temperatures. This study investigated the effects of an increase of 4 °C in soil temperature and a consequent 1-week earlier snowmelt on growth of black spruce [Picea mariana (Mill.) BSP] in the boreal forest of Quebec, Canada. The soil of two natural stands at different altitudes was warmed up with heating cables during 2008-2010 and cambial phenology and xylem production were monitored weekly from April to October. The results showed no significant effect of the treatment on the phenological phases of cell enlargement and wall thickening and lignification. The number of cells produced in the xylem also did not differ between control and heated trees. These findings allowed the hypothesis of a direct influence of soil temperature on stem growth to be rejected and supported the evidence that, in the short term, air temperature is the main limiting factor for xylogenesis in trees of these environments.     

Daily stem water deficit of Norway spruce and European beech in intra- and interspecific neighborhood under heavy drought

High-resolution measurements of stem radius variations provide information about the tree water status with changing climate conditions by swelling and shrinking due to the reduction of xylem water potential and to the exceedance of leaf transpiration over root water uptake. The aim of this study was to analyze daily stem radius variations of Norway spruce and European beech in intra- and interspecific neighborhood. The experimental plots are part of a rainfall exclusion experiment. These variations are species-specific, i.e. spruces have a higher phloem thickness and higher amplitudes during a day than beeches. The amplitudes were significantly higher at the rainfall exclusion plots, but the amplitudes of spruces decreased above 27°C with increasing drought due to reduced transpiration rates and exhausted soil water reserves. The shrinking amplitude was observed for spruces in intraspecific neighborhood from a soil volumetric water content of 0.21?m³?m^?3. In interspecific neighborhood, a shrinking amplitude for spruces could not be observed and revealed a lesser tree water deficit than in intraspecific neighborhood. Beeches showed minor differences with a higher tree water deficit in interspecific neighborhood. Consequently, stem radius variations give insights into a tree's water supply, which could help to understand changes in tree growth.     

Xylogenesis of Pinus pinaster under a Mediterranean climate

Context

The knowledge on cambial activity in water-limited environments, such as the Mediterranean, is still fragmentary. Dendrochronological studies have determined that spring precipitation plays an important part in determining tree-ring width and the properties of tracheids. However, the complex relation between cambial phenology and climate is still far from understood.

Aims

We studied the influence of climate, especially water stress, on maritime pine wood formation with the aim of determining the influence of drought on cambial activity.

Methods

A plantation of maritime pine (Pinus pinaster) was selected in the west coast of Portugal, to monitor cambial activity and wood formation using anatomical observations and band dendrometers. The trees were monitored weekly over 2 years (2010 and 2011).

Results

Xylem differentiation started earlier when warmer late winter temperatures were observed. Water stress triggered an earlier stop of wood formation and also the formation of tracheids with smaller lumen area. In both years a bimodal pattern of stem radial increment was registered by band dendrometers with two periods of increment: one in spring and another in autumn. The xylem anatomy study suggests that the autumnal increment period corresponded mostly to stem rehydration, since the differentiation of new xylem cells by the cambium was not observed.

Conclusion

Maritime pine cambial activity appears to be under a double climatic control: temperature influences cambial onset and water availability growth cessation.     

Anatomical and cellular responses of Pinus monticola stem tissues to invasion by Cronartium ribicola

White pine blister rust (Cronartium ribicola) causes extensive damage to white pines and their associated ecosystems across North America. The anatomical and cellular characteristics of C. ribicola colonization in Pinus monticola branch and stem tissues were studied as a basis for understanding host tree reactions that may be related to resistance. Samples examined showed typical fusiform swelling and some had produced aecia. The reaction of phloem and xylem tissues was compared with non-infected tissue using light and electron microscopy. Cortical parenchyma and phloem polyphenolic parenchyma cells underwent mitotic division, cell swelling, and ca sixfold greater accumulation of phenolic compounds in colonized vs. control stems. In the cortex and secondary phloem, haustoria were common in parenchymatous cells, and hyphae were abundant in the intercellular spaces, but cell death was rare, unless aecia had ruptured the stem cortex. Hyphae were also common in xylem rays, tracheids and between tracheids. Disease-induced changes in the cambial zone included development of cambium-derived xylem traumatic resin ducts. Results demonstrate that diverse host defence responses were activated in the bark of apparently susceptible trees, but lack of mechanical damage by C. ribicola to the phenol-containing host cells and the resin duct system allowed extensive colonization and development of aecia despite elicitation of these stem defences. Interactions between P. monticola and C. ribicola are discussed and compared with other conifer–fungus pathosystems.     

Freezing temperatures in the root zone—effects on growth of containerized Pinus sylvestris and Picea abies seedlings

Roots of 1‐year‐old containerized seedlings of Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.) were experimentally frozen in December. The seedlings were then grown for 3 weeks in a growth chamber and evaluated with regard to root growth capacity (RGC) and shoot elongation. The subsequent RGC of Scots pine declined as root zone temperatures were lowered from ‐6°C to ‐11°C and from ‐11°C to ‐16°C. Almost no root growth was observed after exposure to ‐20°C. Shoot growth was also negatively affected by low root temperatures but less than root growth. Low root temperatures did not affect Norway spruce as much as Scots pine, although root and shoot growth of Norway spruce were reduced after exposure to the lowest test temperatures (‐16°C and ‐20°C). The length of exposure, ranging between 1 and 8 hours had no effect on subsequent growth.     

Cambial Anatomy and Annual Rhythm of Secondary Xylem Development in the Twigs of Azadirachta indica A. Juss. (Meliaceae) Growing in Different Forests of Gujarat State

Abstract

Seasonal behavior of vascular cambium and development of secondary xylem were studied in the 2-3-years-old branches of Azadirachta indica A. Juss. (Meliaceae) growing in moist deciduous (MDF), dry deciduous (DDF) and scrub land (SF) forests of Gujarat State. Development of xylem began with the sprouting of new leaves in January in both MDF and DDF. Xylem production culminated in July in MDF whereas it reached peak in April and July in DDF. Cambial cells ceased to divide with the maturation of leaves in November and October in MDF and DDF, respectively. Mature xylem and phloem derivatives surrounded the cambium in December in MDF and November in DDF. Radial growth in the branches of trees growing in SF was found continuous throughout the year with peak cambial activity in April, July and November. In all the three forests, maximal radial growth was encountered in July when the rains were heavy. Cambial activity and xylem development showed significant correlation with the phenology of the trees in MDF and DDF whereas cambium was found active throughout the year and no correlation was found between the cambial activity and phenology in SF.     

Xylem dysfunction in Yezo spruce (Picea jezoensis) after inoculation with the blue‐stain fungus Ceratocystis polonica

The blue‐stain fungus Ceratocystis polonica is pathogenic to Norway spruce (Picea abies) in Europe, as well as to Yezo spruce (Picea jezoensis) and Sachalin spruce (Picea glehnii) in Japan. The wilting mechanism in P. jezoensis saplings after inoculation with C. polonica was examined based on anatomical studies of the phloem and xylem of periodically harvested trees. In addition, the course of sap ascent in the trunks was traced by injection of acid fuchsin solution at harvest. As an initial external symptom, needle discolouration was observed. In dye conduction tests, xylem dysfunction in the xylem of inoculated trees became obvious. The dehydrated xylem area (dry zone) had extended more than 20 cm above the inoculation wounds, within 1 month after inoculation. When the sap flow to the branches had nearly stopped, the leaves began to discolour. Hyphae of C. polonica colonized the ray tissue around the inoculation wounds, but were absent at the front of the dry zones. Defence reactions occurred in ray parenchyma cells adjacent to the penetrating hyphae. It is suggested that secondary metabolites, which are formed by the ray cells and epithelial cells of resin canals, are involved in the obstruction of sap flow. Limited necrotic lesions of the phloem and cambium were not associated with foliar symptoms. It is proposed that the dry zone formation caused by C. polonica is the main mechanism leading to tree death.