3种木兰科植物叶片SPAD值的分布特征及其与叶绿素含量的关系

为探讨3种木兰科植物灰木莲Manglietia glauca、醉香含笑Michelia macclurei和乐昌含笑Michelia chapensis叶片SPAD值的分布特征及其与叶绿素含量之间的关系,采用SPAD叶绿素计和分光光度法分别测定了三者叶片SPAD值和叶绿素含量.结果表明:3种植物叶片SPAD值的差异主要...     

灵石山米槠(Castanopsis carlessi)林优势种群不同年龄叶片SPAD值的差异及其与叶绿素、氮含量的关系

采用便携式叶绿素仪SPAD-502对灵石山米槠林不同海拔优势种群不同年龄叶片的SPAD值进行野外测定,分别对各海拔不同年龄不同优势种以及各年龄不同海拔不同优势种的叶片SPAD值进行差异性分析,结果表明,灵石山米槠林优势种群叶片SPAD值存在叶龄、种间以及海拔梯度问的差异。对不同海拔米槠林优势种群不同年龄叶片SPAD值与室内分光光度法测定的叶绿素含量以及叶片氮含量进行相关分析,结果表明,SPAD值与叶片单位面积叶绿素及氮含量有显著的相关性。SPAD值是叶绿素含量的真实反映,利用叶绿素仪SPAD-502可以快速有效的诊断种群氮素营养状况。     

Resistance against oxidative stress in leaves of young beech trees grown in model ecosystems with different soil qualities, elevated CO2, and lachnid infestation

Summary The aim of the present study was to investigate whether the resistance of beech foliage(Fagus sylvatica) against oxidative stress was affected by soil quality, nitrogen or CO₂ fertilisation, or lachnid infestation(Phyllaphis fagi). For this purpose young beech trees were grown for four years in reconstructed calcareous or acidic forest soils in open top chambers under ambient or elevated CO₂ concentrations with two levels of nitrogen fertilisation. At harvest lachnid colonisation was observed, preferentially on leaves from trees in calcareous soil and on leaves from trees fertilised with the high nitrogen level. General leaf characteristics such as pigment concentrations, dry mass, and leaf mass ber area were not affected by the soil type, nitrogen fertilisation or CO₂ regime. Leaves colonised with lachnids displayed slightly increased leaf mass per area. When the stress resistance was challenged by exposure to paraquat — a herbicide inducing oxidative stress — leaves from trees grown on calcareous soil maintained significantly longer membrane integrity and, thus, were better protected against stress than leaves from trees on acidic soil. Other experimental variables had negligible or no effects on the resistance against oxidative stress.      

利用 SPAD-502对高山杜鹃氮素营养的快速诊断研究

利用SPAD-502型叶绿素计测定了不同氮水平下高山杜鹃不同生长时期、不同叶位的SPAD 值,分析SPAD值与叶绿素含量和叶片总氮含量的关系。结果表明,不同生长期第1轮和第2轮叶片的SPAD值与叶绿素含量均呈显著线性正相关,其中新梢生长期第1轮叶片的SPAD值与叶绿素含量的相关性最强,而叶绿素各参数中对SPAD值影响最大的是叶绿素b的含量。在不同氮水平下, SPAD值及相对叶色差与单位干重的总氮含量具有显著线性正相关关系, SPAD值的变化能够反映60％的单位干重叶氮含量的变化。因此,可以利用SPAD-502型叶绿素计对高山杜鹃进行快速、无损的氮素营养诊断。     

Can soil plant analysis development values predict chlorophyll and total Fe in hybrid poplar?

Iron chlorosis caused by an elevated soil pH remains an important selection criteria in evaluating hybrid Populus trials in northwest New Mexico. Compared to expensive extraction methods or visual ranking scales, the soil plant analysis development (SPAD) chlorophyll meter is a handheld tool that objectively indicates leaf color. SPAD calibration curves pertaining to Fe status in Populus leaves were developed for two clones: NM-6 (Populus nigra × Populus maximowiczii) and OP-367 (Populus deltoides × Populus nigra). Hybrids were grown in the greenhouse in a sandy loam soil of pH > 8 and the same soil amended with composted biosolids enriched with 420 mg kg⁻¹ Fe. SPAD measurements related to foliar Fe as follows: r ² = 0.72 for NM-6 and r ² = 0.58 for OP-367. SPAD measurements correlated with total chlorophyll as follows: r ² = 0.66 for NM-6 and r ² = 0.85 for OP-367. Based on the total chlorophyll content (supported with foliar element data), Fe sufficiency threshold values from SPAD measurements were estimated in these two clones.     

Shoot biomass growth is related to the vertical leaf nitrogen gradient in Salix canopies

Plant canopy optimization models predict that leaf nitrogen (N) distribution in the canopy will parallel the vertical light gradient, and numerous studies with many species have confirmed this prediction. Further, it is predicted that for a given canopy leaf area, a low vertical light extinction coefficient will promote rapid growth. Therefore, the ideal canopy of fast-growing plants should combine high leaf area index with a low light extinction coefficient; the latter being reflected in a flat vertical leaf N gradient throughout the canopy. Based on data from an experimental Salix stand (six varieties) grown on agricultural land in central Sweden, we tested the hypothesis that shoot growth is correlated with vertical leaf N gradient in canopies of hybrid willows bred for biomass production, which could have implications for Salix breeding. Tree improvement research requires screening of growth-related traits in large numbers of plants, but assessment of canopy leaf N gradients by chemical analysis is expensive, time-consuming and destructive. An alternative to analytical methods is to estimate leaf N gradients nondestructively with an optical chlorophyll meter (SPAD method). Here we provide a specific calibration for interpreting SPAD data measured in hybrid willows grown in biomass plantations on fertile agricultural land. Based on SPAD measurements, a significant and inverse relationship (r(2) = 0.88) was found between shoot biomass growth and vertical leaf N gradient across canopies of six Salix varieties.     

Leaf area of beech (<Emphasis Type="Italic">Fagus sylvatica</Emphasis> L.) from different stands in eastern Austria studied by randomized branch sampling

Few tree size/leaf area correlations have been produced for hardwoods, where the extrapolation from individual branches to the whole tree is less straightforward than in conifers with more regular branching patterns. We used randomized branch sampling to estimate leaf area of European beech (Fagus sylvatica L.) trees of different stands, ages and areas in Austria. Cross-sectional areas (CSA) predicted 87–92% of leaf area variation, the best predictor being the sum of branch CSA. Leaf area was somewhat better correlated with CSA at breast height than at the base of the crown, and using sapwood instead of total CSA made little difference. While there was no effect of growth area, a stepwise regression model showed that dominant trees in pole-stage had, for unclear reasons, significantly higher leaf area/CSA relationships. A comparison with regressions produced from smaller beech trees in other parts of Europe suggests that the leaf area/basal area regression is generally valid for beech in central Europe.     

SPAD-502型叶绿素计在估测大面积区域叶绿素含量中的应用

在每株树木中摘取其上、中、下三部分的叶片,每部分选取5～10片叶子进行测定,对每片叶子的6处不同部位进行SPAD值测定,然后取其平均值作为此片叶子的最终SPAD值,再对所有叶片SPAD值取平均值作为树木的SPAD值。利用GPS技术使树木有了空间信息,并于遥感图像能够结合起来进行分析,为估测大面积区域叶绿素含量创造可能性。结果表明,SPAD-502型手持式测定仪携带方便,测定简单迅速,不损害植被,与GPS技术及遥感反演技术相结合,能简便估测大面积区域叶绿素含量。     

Nutritional effects triggered by the extreme summer 2003 in the free air ozone fumigation experiment at the Kranzberger Forst

Since the year 2000 mature beech and spruce trees were treated in a field experiment with double ambient ozone concentrations. Elevated ozone had no influence on average single leaf biomass and there were also no ozone effects on leaf nutrient concentrations in climatic normal years. However, the extraordinary dry summer 2003 triggered significant differences between the fumigated and control trees. For beech in the year after the drought event the control trees surprisingly had significantly lower foliar levels of K and P than in former years, whereas the ozone exposed trees showed no significant nutritional effects. There are indications, that the trees exposed to double ambient ozone were already adapted to higher ozone values, whereas the control trees experienced extraordinary high ambient ozone concentrations in the dry and sunny summer 2003. For spruce in autumn 2003 and 2004 ozone treated trees had significantly higher foliar levels of K in current year needles than control trees, an effect which cannot be thoroughly interpreted yet on the basis of the dataset available. This article belongs to the special issue „Growth and defence of Norway spruce and European beech in pure and mixed stands“.     

Beech decline in Central Europe driven by the interaction between Phytophthora infections and climatic extremes

During the past decade, and in particular after the wet year 2002 and the dry year 2003, an increasing number of trees and stands of European beech (Fagus sylvatica L.) in Bavaria were showing symptoms typical for Phytophthora diseases: increased transparency and crown dieback, small‐sized and often yellowish foliage, root and collar rot and aerial bleeding cankers up to stem heights of >20 m. Between 2003 and 2007 134 mature beech stands on a broad range of geological substrates were surveyed, and collar rot and aerial bleeding cankers were found in 116 (86.6%) stands. In most stands the majority of beech trees were declining and scattered or clustered mortality occurred. Bark and soil samples were taken from 314 trees in 112 stands, and 11 Phytophthora species were recovered from 253 trees (80.6%) in 104 stands (92.9%). The most frequent species were P. citricola, P. cambivora and P. cactorum. Primary Phytophthora lesions were soon infected by a series of secondary bark pathogens, including Nectria coccinea, and wood decay fungi. In addition, infected trees were often attacked by several bark and wood boring insects leading to rapid mortality. Bark necroses were examined for their probable age in order to determine whether the onset of the current Phytophthora epidemic was correlated to rainfall rates recorded at 22 Bavarian forest ecosystem monitoring stations. A small‐scale survey in nine Bavarian nurseries demonstrated regular infestations of all beech fields with the same range of Phytophthora species. The results indicate that (1) Phytophthora species are regularly associated with beech decline and may also be involved in the complex of ‘Beech Bark Disease’, (2) excessive rainfalls and droughts are triggering the disease, and (3) widespread Phytophthora infestations of nursery stock might endanger current and future silvicultural projects aiming on the replacement of non‐natural conifer stands by beech dominated mixed stands.     

Induced response of the Siebold's beech (Fagus crenata Blume) to manual defoliation

Siebold's beech (Fagus crenata) was manually defoliated for two successive years. The beech caterpillar (Quadricalcarifera punctatella) was used in a bioassay to determine insect performance. Survival and body size were low on foliage from defoliated trees. Reduced foliar nitrogen and increased tannin content were probably the main causes of the low insect performance. Leaves were less tough on defoliated trees than in controls. Two sucessive years of manual defoliation caused stronger induced resistance than one year defoliation. The quality, as well as the quality of the foliage, decreased the year following manual defoliation; total weight of leaves on a tree was less than one half of that before treatment. Severe defoliation may cause a decrease of leaves the following year and starvation may limit populations. Delayed induced resistance of beech trees is proposed as a possible cause of the cyclical population dynamics ofQ. punctatella. The delayed induced response also affected folivorous insects other thanQ. punctatella.     

Influence of nurse birch and Scots pine seedlings on early aerial development of European beech seedlings in an open-field plantation of Central France

Beech woodland can be restored by direct planting of beech (Fagussylvatica L.) seedlings in abandoned areas, but this methodis generally avoided in forestry because of the growth difficultiesof beech in full-light conditions. This study tested a methodthat consists of planting beech seedlings in full-light conditionswith silver birch (Betula pendula Roth.) or Scots pine (Pinussylvestris L.) as nurse trees. A total of 65 two-year-old beechseedlings surrounded by either 3, 4, 5 or 6 pine or birch seedlingsor without competitors were planted in bare-soil open-fieldconditions in Central France. Tree growth and light availabilitywere monitored at the end of each of the following 3 years,and soil moisture was also measured the last year. At the endof the experiment, aerial biomass measurements were made onthe beeches. Results showed that relative beech growth in termsof diameter was significantly reduced by the local neighbouringtrees and that this reduction was particularly noticeable inthe pine treatments where there was extensive pine seedlingdevelopment. In contrast, relative beech height did not exhibitany significant variations among the treatments. Light availabilitywas decreased by the neighbouring trees, especially in the pinetreatments. Light reduction was more pronounced in the last2 years and in the middle or lower parts of the canopy of thesubject beech. Soil water content was lower under the pine canopiesthan under the birch canopies, and was positively correlatedto beech relative diameter growth. Specific leaf area, height-to-diameterratio and crown length-to-crown width ratio weakly but significantlyincreased with competition from the neighbours. Beech biomasspartitioning was only weakly affected by the treatments. Thisstudy showed that nurse trees tend to reduce beech growth butimprove form although effects remained weak due to the insufficientlength of the experiment. Further studies are also needed toquantify the effects of the neighbourhood on the growth of competitiveherbaceous vegetation and on changes in the microclimatic conditions.     

Changes in the relationship between tree size and aboveground respiration in field-grown hinoki cypress (Chamaecyparis obtusa) trees over three years

Respiration measurements of aerial parts of 18-year-old hinoki cypress (Chamaecyparis obtusa (Sieb. et Zucc.) Endl.) trees were made under field conditions over three years to study changing relationships with tree age between respiration and phytomass, phytomass increment, and leaf mass. The relationship between annual respiration (r(a)) and phytomass (w(T)) was approximated by a proportional function (r(a) = aw(T)), where the proportional constant (a) decreased year by year. The effect of time on the relationship between annual respiration and phytomass of each sample tree was fitted by a power function. Respiration of the tree suppressed by the canopy decreased year by year, but respiration of the other trees increased slightly with age. The relationship between annual respiration and leaf mass was also approximated by a generalized power function. Excluding the suppressed tree, the relationship between annual respiration (r(a)) and the annual increment of aboveground phytomass (Deltaw(T)) was described by a proportional function (r(a) = 2.27Deltaw(T)), where the proportional constant, 2.27, was independent of sample tree and year, indicating that about 2.3 times of the annual aboveground phytomass increment equivalent was respired annually. For any tree, the time constant relationships between annual respiration and leaf mass and phytomass increment for different-sized trees were similar to the corresponding time continuum relationships. In contrast, the time continuum relationship between annual respiration and phytomass differed from the time constant relationship, indicating that respiration of less active woody tissue contributed significantly to aboveground respiration. Based on the relationship between tree size and annual respiration, annual aboveground stand respiration was estimated to be 25.0, 26.9, and 25.8 Mg(dm) ha(-1) year(-1) for the three consecutive years, respectively, and the corresponding aboveground stand biomass was 60.0, 69.0, and 76.8 Mg(dm) ha(-1).     

Impact of late frost events on radial growth of common beech (Fagus sylvatica L.) in Southern Germany

Phenological, temperature, and tree-ring data were used in order to identify and quantify the impact of late frosts on common beech (Fagus sylvatica L.) at different altitudes in Southern Germany during the last century. For this intention, dendroecological investigations were made upon trees at the Meteorological Observatory Hohenpeißenberg as well as from seven stands in the Bavarian Forest and 17 stands at the northern fringe of the Alps. From these locations, a considerable number of severe growth minima in the tree-ring series could be related to late frost in the days of or immediately after leaf unfolding. The frequency of frost-related growth minima increases with altitude. In individual years, radial growth can be reduced by more than 90% (stand mean) in relation to the average growth of the ten previous years. Hence, late frosts are considered as important ecological events that strongly affect beech vitality and competitiveness especially at high altitudes. Evidence of significant impacts on radial growth by late frosts distinct before leaf unfolding or with temperatures above ?3°C was not found. Also, increasing frequency and intensity of late frosts during recent decades were not ascertained. Hence, the recently observed decreased vitality of common beech accompanied by growth depressions especially at high altitude sites in Central Europe cannot be explained as a consequence of late frost damage.     

Phenological phases of common beech (Fagus sylvatica L.) and their dependence on region and altitude in Southern Germany

Long-term observational data of the phenological network of the German Weather Service (DWD) were used to derive the altitudinal dependence of leaf unfolding (LU), leaf discolouration (LD) and the length of the vegetation period (LD-LU) of common beech (Fagus sylvatica L.) in different regions of Southern Germany. Depending on altitude, the vegetation period starts in Bavaria between the 2nd and 24th May with a delay of around 2 days per 100 m altitude increase (averaged over the last 30 years). In Southern Bavaria, LU starts a few days earlier as well as increases slightly faster into higher altitudes than in the northern regions. This can be explained by a more favourable supply of warmth and radiation in the south. The end of the vegetation period, defined here as the beginning of LD, showed no altitudinal dependence, which reflects an interaction of endogenous and exogenous factors as triggers for LD. In the Bavarian Forest region, altitudinal dependences are strongly influenced by sites which are affected by cold-air tailback (‘Kaltluftstau’) was obtained. Hence, phenological gradients should only be derived and applied above a phenological inversion zone. The calculated vertical gradients for LU can be used together with available observational data to estimate the beginning of the forest vegetation period of beech for different altitudes of Bavaria.     

Comparative studies on the fine root status of healthy and declining spruce and beech trees in the Bavarian Alps and occurrence of Phytophthora and Pythium species

Investigations on root and crown status of spruce and beech were carried out on selected trees in the ‘Werdenfelser Land’ area (Bavarian Alps, Southern Germany). In addition, the association of fine root pathogens of the genera Phytophthora and Pythium with the trees’ rhizosphere was studied. In a variety of stands representing various site conditions, soil and root samples were taken from 12 spruce (Picea abies) pairs and eight beech (Fagus sylvatica) pairs. Each pair consisted of a healthy and a declining tree as indicated by crown transparency. The root status was characterized using a set of parameters, and correlations between crown and fine root status were observed. In spruce, most parameters decreased significantly with increasing crown transparency, whereas in beech, correlations were less pronounced. The total number of lateral roots per cm small root (diameter 2–5 mm) was significantly lower in both species for declining trees compared to healthy trees. Pythium spp. were isolated from 15 of 24 soil samples taken from under spruce, and from eight of 16 samples from under beech. Phytophthora citricola was found in two beech stands only. Among the isolated species, Pythium anandrum, Pythium inflatum and Pythium acrogynum were identified according to morphological features. After polymerase chain reaction‐restriction fragment length polymorphism analysis, residual Pythium isolates were assigned to four different groups. No crown transparency‐dependent differences in isolation frequency were found. In soil infestation tests, all species tested caused root damage on both young spruce and beech plants, with P. citricola being the most aggressive pathogen. Additionally, Pythium‐infected beeches showed severe leaf chloroses and necroses. Due to their low isolation frequency, Phytophthora spp. are not considered to play a major role in the decline of spruce and beech in the investigated area. Pythium spp., however, were isolated frequently, showed pathogenicity towards the fine roots of spruce and beech, and are therefore considered to be at least contributing factors in the decline of Bavarian mountain forests.     

Radial distribution of carbohydrate reserves in the trunk of declining European beech trees (Fagus sylvatica L.)

Context

European beech (Fagus sylvatica L.) is considered threatened by anticipated climate change, but the physiological causes of potential beech decline or mortality remains poorly understood.

Aims

The purpose of the present study was to fuel debate about the assumption that carbohydrate depletion is involved in the decline of mature European beech.

Methods

The health status of beech trees from a severely declining stand was visually assessed by examining their crown condition. Content and radial distribution of non-structural carbohydrates (starch and soluble carbohydrate) were analyzed in the trunks and compared to those reported earlier in trunks of healthy beech trees.

Results and discussion

The distribution of carbohydrate in the beech trunks recorded here seemed affected by decline. We found a stronger radial decrease of starch content than those reported earlier for healthy beech trees. Carbohydrate reserves appear partially maintained in the outermost rings while starch depletion occurred in older wood rings in declining trees that may be able to mobilize carbohydrate reserves from older wood rings in response to successive climatic constraints.     

葡萄叶片SPAD值与测定位置及光合色素含量的关系

为了实时、准确、无损地检测葡萄叶片中叶绿素的含量水平,以评价葡萄生长发育与营养状况,确保栽培的科学和施肥的合理,以‘夏黑’‘巨玫瑰’‘户太8号’‘阳光玫瑰’‘金手指’‘摩尔多瓦’这6个品种葡萄叶片为试验材料,采用叶绿素测定仪测定了6个葡萄品种叶片不同位置的SPAD值,使用分光光度计测定了葡萄叶片光合色素含量,探寻了葡萄叶片SPAD值的最适测量位点,分析了SPAD值与光合色素含量的关系,并建立了SPAD值与光合色素含量的线性函数拟合方程。结果表明:6个品种葡萄叶片的SPAD值,叶片顶部的均最高,中部的均次之,基部的均最小;其中,葡萄叶片中部的SPAD值最具有代表性。6个品种葡萄叶片的SPAD值与叶绿素a、叶绿素b、总叶绿素和类胡萝卜素含量均显著相关,其与叶绿素a含量的相关性最高;在不同品种葡萄叶片的SPAD值与叶绿素a、叶绿素总含量、类胡萝卜素含量的线性函数拟合方程中,‘户太8号’的拟合方程决定系数值最高,其叶绿素a、叶绿素总含量、类胡萝卜素含量的决定系数值分别为0.903 5、0.867 2、0.749 9;在叶绿素b与葡萄叶片SPAD值的拟合方程中,‘金手指’的拟合方程为y=0.034 7x-0.084 0,其决定系数R2值(0.773 4)最高。单一品种葡萄叶片的SPAD值与光合色素含量的回归模型的拟合准确率比6个葡萄品种汇总拟合回归模型的要高。     

The emergence of beech leaf disease in Ohio: Probing the plant microbiome in search of the cause

American beech (Fagus grandifolia) is the target of a newly emerging disease in North America called beech leaf disease (BLD) that affects and disfigures leaves and which can lead to tree mortality. Beech leaf disease may be caused by a newly recognized subspecies of the anguinid nematode Litylenchus crenatae subsp. mccannii, but the associations of this nematode with bacterial and fungal taxa are unknown. We examined microbial communities associated with beech leaves affected by BLD in a 16‐year‐old American beech plantation using molecular methods. We detected L. crenatae subsp. mccannii in anywhere from 45% to 90% of leaves depending on the degree of visual BLD symptoms. Approximately 37% of asymptomatic leaves contained L. crenatae subsp. mccannii, whereas 90% of buds associated with symptomatic leaves contained L. crenatae subsp. mccannii. We found that fungal communities on leaves and buds were unaffected by BLD, but bud and leaves had significantly different fungal communities. Bacterial communities on buds also were unaffected by BLD, but bacterial communities were significantly different between symptomatic and asymptomatic leaves suggesting that the nematode could be altering the community of bacteria on the leaves. Clone libraries indicate that Wolbachia, an intracellular endosymbiont of arthropods, was found only on symptomatic leaves and buds associated with either symptomatic or asymptomatic leaves. In addition, only symptomatic leaves contained taxa in the genus Mucilaginibacter, which previous studies suggest could produce exopolysaccharides. These bacterial taxa could represent a marker for the vector of L. crenatae subsp. mccannii that enables spread between trees and a possible endosymbiont that could facilitate nematode feeding and establishment on nematode infested leaves. Our results are the first to examine changes to the leaf microbiome of this newly emerging pest and may aid identification of mechanisms associated with the spread and success of L. crenatae subsp. mccanni.     

The effect of bagging branches on levels of endophytic fungal infection in Japanese beech leaves

The species composition of the endophytic mycobiota in leaves of Japanese beech trees (Fagus crenata) and the sources for leaf infections were studied in a forest reserve situated in central eastern Honshu, Japan. To clarify the mechanism of infection of leaves, half of the branches were covered with polyethylene bags and species composition and levels of endophytic fungal infection were then compared with those of unbagged controls. Isolations were carried out from the leaves, petioles, and current‐year twigs of both, bagged and unbagged branches. Additionally, species composition was detected in overwintered terminal buds of beech trees and in the leaves of potted seedlings that had been placed in the field in different seasons. The species assemblage of the unbagged leaves, petioles, and current‐year twigs was dominated by Mycosphaerella buna, Ascochyta fagi, Periconiella sp., and Tritirachium sp. Other frequently recovered species were Xylaria sp., Phomopsis sp., and Tubakia dryina. Mycosphaerella buna and A. fagi were never isolated from leaves on bagged branches. A. fagi was, however, detected on both bagged and unbagged petioles and current‐year twigs at comparatively low isolation frequencies. The detection of Periconiella sp. on all occasions in both bagged and unbagged leaves was a characteristic feature that differs from those of the other three dominant endophytic fungi. The fungus was also detected without significant differences in bagged and unbagged petioles and current‐year twigs on most sampling dates. Furthermore, Periconiella sp. was isolated from immature twigs inside the bud scales. Tritirachium sp. was frequently detected in unbagged leaves and petioles and in both bagged and unbagged current‐year twigs, and rarely in bagged leaves and petioles, but was never recovered from terminal buds. The results of the potted seedling experiments revealed that all four dominant species had airborne inocula. The infection of leaves by M. buna occurs exclusively by airborne propagules, i.e. ascospores in spring and conidia in autumn. In Periconiella sp. hyphal growth of the fungus from immature twigs inside the buds into the leaf tissues was suggested in addition to infection by airborne inocula. Tritirachium sp. hyphae were suggested to grow from previous‐ to current‐year twigs. Ascochyta fagi was present in the outermost scales of overwintered terminal buds, but no systemic growth of the fungus into the petioles and current‐year twigs was observed. Our technique of covering the branches before new leaves unfolded was effective in preventing infection by airborne inocula of endophytic fungi.