Untersuchungen zur Biologie,Morphologie und Verbreitung von Herpotrichia parasitica (Hartig) E. Rostrup (vormals Trichosphaeria parasitica Hartig)1

Studies on thc biology, morphology ancl clistribution of Herpotrichin parasitica (Hartig) E. Rostrup (Trichosphaeria parasitica Hartig) 1. Distribution and morphology. H. paratsitica (Hartig) E. Rostrup (Impcrfcct stage Pyrcnochaeta parasitica Freyer ct van der Aa) occurs in Southern Bavaria on European silver fur. Thc Fungus produces thick cushions of mycelium on the under sidcs of shoots and ovcr thc stomatal rows on needles, penctrates bark tissue as well as necdlcs and causes dcath of needles. Spores of both stagcs gcrminate vcry well in water under various conditions; pycnosρores germinated on silvcr fir needles undcr laboratory Conditions.     

Über Pyrenochaeta parasitica spec. nov., die Nebenfruchtform von Herpotrichia parasitica (Hartig) E. Rostrup (= Trichosphaeria parasitica Hartig

Pyrenochaeta parasitica spec. nov., the conidial stage of Herpotrichia parasitica (Hartig) E. Rostrup (= Trichosphaeria parasitica Hartig). This study contributes to the systematic position of the fungus Herpotrichia parasitica (Hartig) E. Rostrup, found on the needles of silver fir (Abies alba Mill.) and describes its Pyrenochaeta conidial stage.     

Note sur l'aptitude lignivore du Lachnellula willkommii (Hartig) Dennis,agent du chancre du mélèze

Note on the wood destroying capacity of Lachnellula willkommii (Hartig) Dennis, the larch canker fungus. The wood destroying capacity of Lachnellula willkommii in larch was studied in vitro using a method based on loss of weight. Two species of larch and their hybrid were used. The behaviour of the fungus on sapwood and heartwood differed.     

Epidemiological role of strains of Cryphonectria parasitica isolated from hosts other than chestnut

Cryphonectria parasitica is not common in the forests of the Emilia Romagna region of Italy on hosts other than chestnut. Three isolates with morphological cultural characteristics of C. parasitica were collected: two from infected bark of Quercits pubescens (EQ1 and EQ2) and one from Carpinus betulus (EC1). Pathogenicity tests on chestnut and crosses in vitro with strains of C. parasitica of different mating types confirmed that EQ1 and EQ2 are C. parasitica isolates, whereas EC1 seems to belong to a different species. Progeny analysis of crosses with the isolate EQ1 and a strain from chestnut showed the formation of new vegetative compatibility groups. Therefore hosts other than chestnut can contribute to increasing pathogen variability through recombination of the v-c genes.     

Long‐term field study of transgenic hypovirulent strains of Cryphonectria parasitica in a forest setting

Transgenic hypovirulent strains of Cryphonectria parasitica, the chestnut blight fungus, engineered to contain a chromosomally integrated full‐length infectious cDNA copy of virulence‐attenuating hypoviruses, differ from natural hypovirulent strains in the ability to transmit hypoviruses to ascospore progeny and with 100% efficiency through asexual spores. We report the results of a long‐term field study that examined whether these properties result in enhanced hypovirulence establishment, dissemination and persistence under field conditions. Informed by previous field results using a severe hypovirus, this study that employed 144 American chestnut trees was designed to provide improved inoculum formulation and delivery and to include the use of a mild hypovirus isolate (less debilitating) CHV‐1/Euro7 in an attempt to increase dissemination. Isogenic transgenic hypovirulent (TG), non‐transgenic cytoplasmic hypovirulent (CH) or virus‐free virulent (V) treatment strains were applied to artificially initiated and natural C. parasitica cankers three times each year for 7 years. Reservoirs of treatment inoculum also were initiated and refreshed annually for the first 6 years of the study. Sampling of 111,000 individual ascospores from 4,500 perithecia confirmed hypovirus‐containing spermatia successfully transmitted TG hypoviruses to ascospore progeny under field conditions. Surprisingly, TG ascospore progeny were recovered 3 years after the last annual application of treatment inoculum. Repeated sampling of over 440 cankers revealed dissemination of both CH and TG hypovirulent strains. However, no significance differences in establishment or dissemination were observed for the two hypovirulent strains. The results are discussed in terms of the contribution of ascospore progeny to infection, competition by endemic virulent C. parasitica, size of inoculated trees and the biological control potential of TG hypovirulent strains.     

Diversity of Cryphonectria parasitica in callused chestnut blight cankers on European and American chestnut

Infection of American and European chestnuts with the chestnut blight fungus Cryphonectria parasitica results in the formation of cankers, lesions caused by the growth of mycelia within bark tissue of the host plant. Infection of the fungus with Cryphonectria hypovirus 1 (CHV‐1) results in conversion of the mycelial phenotype from virulent to hypovirulent, thus allowing production of callus around cankers as a reaction by infected trees, rendering active into inactive cankers. In this study, we sampled one USA and six European chestnut stands and assessed frequency of hypovirulent C. parasitica and diversity of vegetative compatibility (vc) types present in calluses and randomly sampled cankers. Callused cankers on C. dentata at West Salem in the USA yielded significantly more hypovirulent C. parasitica isolates compared with four sampled populations on C. sativa, while all six sampled European populations did not show any statistically significant differences among themselves. We observed no correlation between hypovirulence frequencies in randomly sampled cankers and calluses, as well as no correlation of C. parasitica vc type diversity in calluses and residential populations of the fungus. Furthermore, even though we have observed calluses with more than one vc type, they do not occur regularly. Even when present in C. parasitica populations with high vc type diversity, no more than three different vc types were observed in a single callus.     

Seasonal effect on infection and development of lesions caused by Cryphonectria parasitica in Castanea sativa

Seasonal variation in the development of chestnut blight, caused by Cryphonectria parasitica, was investigated by inoculating in situ chestnut trees and in vitro excised chestnut segments, at either monthly or 3‐monthly intervals throughout 30 months. Inoculations were made with conidia and mycelium of a virulent isolate and with mycelium of a hypovirulent isolate. Conidial inoculations of living sprouts or excised segments between May and July resulted in the greatest incidence of infection whereas inoculations in autumn and winter, in vitro as well as in situ, did not reveal any visible disease. However, from these symptomless inoculated stems, C. parasitica was isolated 3 months after inoculation. Inoculations with the mycelium of the virulent isolate always resulted in lesions, except in January 1999, and the greatest rate of lesion development occurred for inoculations made in the spring and summer. There was a significant seasonal effect on lesion development. Lesions caused by the hypovirulent isolate, smaller than those caused by the virulent isolate, followed a similar seasonal pattern. The same seasonal variations were observed for inoculations in vitro of excised segments. Relative water content (RWC) of chestnut bark significantly varied with bark sampling date. The rate of lesion development in sprouts significantly correlated with average minimum (ATn) and maximum (ATx) temperatures and the sum of rainfall during inoculation period, with the rate of lesion development measured in excised segments 10 days after inoculation (R10d) and with RWC measured on the day of inoculation. In multiple regression models, variables ATx and R10d best explained variation in lesion development.     

Cryphonectria parasitica in Portugal: diversity of vegetative compatibility types,mating types,and occurrence of hypovirulence

Cryphonectria parasitica, the causal agent of chestnut blight, has been responsible for the decline in chestnut in Portugal for the last two decades. In order to improve understanding of C. parasitica diversity, a total of 617 isolates from all affected chestnut‐growing areas in continental Portugal, Madeira and the Azores archipelagos were studied. Nine vegetative compatibility (vc) types were identified among the isolates. EU‐11 was the most widespread vc type comprising 80.2% of the isolates, followed by EU‐12 (7.1%) and EU‐66 (6.6%). Two of the Portuguese vc types could not be assigned to a known European vc type. The diversity of vc types was low in the Portuguese populations of C. parasitica, but comparable with other areas where C. parasitica was introduced recently. The frequent occurrence of perithecia and both mating types of C. parasitica indicates that sexual reproduction of the chestnut blight fungus is common in Portugal. One C. parasitica isolate from Trás‐os‐Montes showed a white culture morphology and contained dsRNA, indicating the presence of hypovirulence in this area.     

Synergistic effect of drought and chestnut blight (Cryphonectria parasitica) on growth decline of European chestnut (Castanea sativa)

The chestnut blight fungus [Cryphonectria parasitica (Murill) Barr] has threatened European chestnut stands (Castanea sativa Mill.) in the 20th century, but infected trees recovered because of the appearance of hypovirulent strains. However, within a dry inner Alpine valley (Italy), blight‐infected C. sativa showing various degrees of crown dieback and dead trees were found. We conducted a dendroecological analysis to retrospectively evaluate a possible synergistic effect of blight infection in the early 1970s and climate stress on the growth decline of C. sativa. In the Eisack Valley (Italy), where annual precipitation is <700 mm, increment cores were taken from blight‐infected C. sativa (n = 103) showing different levels of decline symptoms, i.e. extent of crown dieback (healthy, moderate, severe), and from dead trees. Ring width and basal area increment (BAI) chronologies were developed based on dendroecological methods. Growth–climate relationships were explored using response function analysis and Pearson correlation coefficients. Major findings of our study were: (i) C. sativa growth is limited by low precipitation from December to February and high temperatures in June, (ii) BAI of all vitality classes except healthy trees shows a decreasing trend since 1980; and (iii) a severe drought in 1996 accelerated growth decline and caused death of infected C. sativa individuals. Because of the strong influence of climate on radial tree growth within the study area and observed divergent growth trends in selected vitality classes after infestation by hypovirulent C. parasitica strains in the early 1970s, we conclude that although blight infection is a prerequisite for observed growth declines, soil water availability in the years of drought strongly affects susceptibility to tree death.     

Distribution and severity of damage by Cryphonectria parasitica in the chestnut stands in Guilan province,Iran

Chestnut blight caused by Cryphonectria parasitica has recently been reported from Guilan province, the only region with natural chestnut (Castanea sativa) stands in Iran. During the past few years, chestnut stands in Iran have been seriously threatened by the fungus, incidence of the disease is increasing and vast canopies are reduced to sprouts. As yet, there is no report on the disease distribution and severity in this region. Six sites from three main growing regions of chestnut in Guilan province were selected for investigation. We report occurrence and evaluation of the damage of the disease caused by C. parasitica. To evaluate the scale of damage, the investigated trees were classified into six categories based on the degree of crown damage, the number of canker wounds and the presence of the fungus. Index of health condition was calculated for all sites. During this study, a total of 250 isolates of Cryphonectria species were obtained, of which 232 isolates were C. parasitica and 18 were Cryphonectria radicalis. Castanea parasitica was observed in all regions. Index of health condition was scored from 0 to 6 with 0 being disease free and six being the most severe infection. Index results in the investigated sites varied between 0.69–5.45 and 0.93–5.55 for years 2006 and 2007, respectively. The highest damage was found in Doran (I_H = 5.55), which is located some 100 km away from Shahbalutmahalleh, the site with lowest damage (I_H = 0.93). This is the first extended report on aspects of chestnut blight in Iran.     

Mechanisms and metabolomics of the host–pathogen interactions between Chestnut (Castanea species) and Chestnut blight (Cryphonectria parasitica)

Chestnut blight is a stem‐girdling disease of Castanea caused by the fungal pathogen Cryphonectria parasitica. Chestnut blight affects all Castanea species to some degree. In Asian species, chestnut blight is a commercially relevant disease which primarily affects nut production. In American and European species, chestnut blight has caused significant declines in wild populations and continues to negatively affect nut production in the European chestnut (C. sativa). Despite the profound effect of this disease in the Castanea genus, very little is known concerning the factors involved in the host–pathogen interaction between C. parasitica and its Castanea hosts. This review summarizes information on known mechanisms and metabolites involved in the host–pathogen interaction and contributes original information on the pathogen in relation to susceptible and putatively resistant genotypes with a view to furthering research that will promote a better understanding of this devastating disease and enable its control.     

Growth and physiological responses of chestnut calli to crude extracts of virulent and hypovirulent strains of Cryphonectria parasitica

The effect of crude extracts (CEs) of virulent (V‐E4) and hypovirulent (H‐E13) Cryphonectria parasitica strains on growth and physiological activities of chestnut calli was investigated on cell cultures obtained from either a susceptible Castanea sativa or a resistant Castanea mollissima chestnut species. The V‐E4 CEs significantly reduced calli fresh weight in both species when used at 50 and 500 μg g^–1 and, in general, showed an inhibitory effect on either O₂ uptake and H⁺‐ATPase activity. Only the highest (500 μg g^–1) concentration of H‐E13 CEs had a detrimental effect on callus growth, whereas the lowest one (5 μg g^–1) induced a slight, but significant early increase in callus fresh weight of both genotypes. The O₂ uptake and H⁺‐ATPase activity were enhanced by the presence of H‐E13 CEs. The decrease of callus growth and physiological activities, caused by V‐E4 CEs, was attributed to toxic compounds produced by the C. parasitica virulent strain, whereas the stimulatory effects of H‐E13 CEs on the overall callus metabolism seemed to be due either to the lack or very reduced amount of phytotoxic compounds and to growth‐regulating substances produced only by the hypovirulent strain of the fungus.     

Growth and ultrastructural modifications to chestnut calli induced by culture filtrates of virulent and hypovirulent Cryphonectria parasitica strains

Callus cultures of two susceptible Castanea sativa cultivars, ‘Garrone rosso’ and ‘Clone 71’ were grown on culture medium supplemented to 50% with culture filtrates (CFs) from E4 virulent (E4-V) or E13 hypovirulent (E13-H) Cryphonectria parasitica strain, respectively. E13-H CFs caused a reduction in fresh weight and an increase in dry weight on calli of both genotypes. E4-V CFs did not induce any modification of these parameters compared with the control, but fresh weight of ‘Clone 71’ was reduced. Light and electron microscopy observation showed the presence of large osmiophilic aggregates in the vacuoles and, sometimes, the ruptured tonoplast in the E4-V treated calli. Calli grown on El 3-H CFs had a meristematic-like appearance, with small, scarcely vacuolated cells containing altered mitochondria and plasmalemma proliferations.     

Rindenkrebs der Eßkastanie (Cryphonectria parasitica) in Südwestdeutschland

Chestnut blight (Cryphonectria parasitica) in South-west Germany . In 1992 and 1993, chestnut blight (Cryphonectria parasitica) was found in three stands in the south-west of Germany. Each of the three stands has only one compatibility group (vc-group). Two of them (97 and 75 Isolates) belong to the vc-group I. They are compatible with the Swiss vc-group I. The 43 Isolates from the third stand were not compatible with the most common Swiss vc-groups (vc-group I-V). In the German vc-group I, one hypovirulent strain was found.     

Chestnut blight (Cryphonectria parasitica) north of the Swiss alps

Chestnut blight (Cryphonectria parasitica) was found in several stands in the Swiss plateau. At each site only one or two compatibility (vc) groups were present. The vc-group I was the most common. At a heavily infested site also young oaks (Quercus petraea) were infected.     

Low genetic diversity but frequent sexual reproduction of the chestnut blight fungus Cryphonectria parasitica in Azerbaijan

Chestnut blight was first recorded in Azerbaijan on the native European chestnut (Castanea sativa) in 2004, and since then, the disease is expanding in the country. In this study, chestnut blight was detected in seven of eight forest districts that were inspected. To characterize the local population structure of the chestnut blight fungus, 199 Cryphonectria parasitica isolates were collected and assessed for vc type, mating type and microsatellite genotype. Among these isolates, one dominant vc type was detected comprising 93% of the isolates. Six additional vc types were identified among the other isolates. The microsatellite analysis revealed a very similar pattern with 96.5% of the isolates belonging to the same multilocus genotype. Both mating types of C. parasitica were present in all seven districts with a mating type ratio not different from 1:1 in five districts. In accordance with the occurrence of both mating types, sexual fruiting bodies (perithecia) of C. parasitica were found in all districts with an overall prevalence on 20.6% of the cankers. The dominant vc type and microsatellite genotype in Azerbaijan do not occur in Europe, but have previously been found to be widespread in neighbouring Georgia. Our study reveals that sexual reproduction in the invasive C. parasitica population in Azerbaijan is frequent, although the population shows a low genetic diversity. This could favour the biological control of chestnut blight using hypovirulence, which so far does not seem to be present in Azerbaijan.     

Distribution of chestnut blight and diversity of Cryphonectria parasitica in chestnut forests in Bulgaria

We surveyed chestnut stands at 18 sites in 11 locations in Bulgaria in 2005 and 2007 for the presence of chestnut blight. We found chestnut blight in seven locations (Belogradchik, Berkovitsa, Brezhani, Barziya, Govezhda, Petrich and Petrovo) but not in four others (Tsaparevo, Kresna, Dupnica and Botevgrad). We successfully isolated Cryphonectria parasitica from cankers on 606 trees with symptoms of chestnut blight and assayed them for vegetative compatibility (vc) types and mating type. Three vc types were identified among the 606 isolates; all three were among the European vc types with known vegetative incompatibility (vic) genotypes. Vc type EU‐12 was the most common, representing 80% of the isolates, and was found at all locations with blight, with the exception of Belogradchik in north‐west Bulgaria, where all isolates were vc type EU‐2. Only one population (Barziya) had more than one vc type, with a combination of EU‐12 and EU‐10 in almost equal frequencies. Similarly, the diversity of mating types was very low. All but three of 536 isolates assayed were in mating‐type MAT‐1; MAT‐2 was only found in one population in the north‐west (Berkovitsa). We inspected 671 bark samples from chestnut blight cankers with stromata of C. parasitica and found perithecia in only 33, of which 28 were from Berkovitsa where MAT‐2 was present. We did not detect hypoviruses in any of the 270 isolates screened using the standard double‐stranded RNA extraction protocol. Similar to results from previous studies in south‐eastern Europe, the diversity of vc types and mating type of C. parasitica in Bulgaria is low, and reproduction of the fungus is mainly asexual. Unfortunately, naturally occurring hypovirulence was not detected. Nevertheless, we observed a small number of superficial cankers typical of those caused by C. parasitica isolates infected with a hypovirus.     

落叶松锉叶蜂为害的松林光谱特征分析

通过对不同程度危害的落叶松反射光谱进行测定,研究了不同程度危害的落叶松在绿光区、红光区和近红外区反射光谱的变化特征,并对光谱反射曲线进行微分分析。结果表明,绿光区、红光区和近红外区的落叶松光谱反射率随危害程度的加重分别呈现下降、上升和下降的趋势;对反射率曲线进行微分分析,健康落叶松、轻度、中度和重度为害后的一阶导数光谱反射率最大值随着危害程度增加而下降,并且向短波方向移动(蓝移)。实测光谱数据提取的归一化植被指数(NDVI)与落叶松锉叶蜂危害程度呈显著负相关,对应用遥感技术早期监测落叶松锉叶蜂灾害具有重要指示意义。     

Eutypella parasitica naturalised in Bohemian and Polish Silesia

In 2015 and 2016, the North American maple tree pathogen Eutypella parasitica was identified in Bohemian and Polish Silesia. To date, the pathogen has been recorded in 35 locations over an area of approximately 400 km². It has been found primarily in natural stands in Silesia in several types of ravine and alluvial forests, in other natural or commercial forests, in riparian stands and in open landscapes. The proportion of diseased maples ranged between approximately 1% and 50% in certain locations. Acer pseudoplatanus was confirmed as the main host but infections were also detected on A. platanoides and A. campestre. E. parasitica damages the most valuable sycamore timber and represents a clear risk for maple cultivation in the region.