Fruiting and sporulation of Thekopsora and Chrysomyxa cone rusts in Picea cones and Prunus leaves

Natural fruiting and sporulation of cone rusts were investigated in cones of Picea spp. and leaves of Prunus spp. in a botanical garden in northern Finland in 2007–2012. Thekopsora areolata was the most frequent cone rust in Picea abies cones, where it colonizes the host tissues and hinders normal seed development. Aecia of T. areolata were also common in cones of Picea engelmannii and occasionally in cones of P. glauca. Aecia of T. areolata sporulated in cones that were at least one year old. Chrysomyxa pirolata, another pathogenic cone rust, fruited and sporulated annually but infrequently in current‐year cones of P. abies. The spruce needle rust, Chrysomyxa ledi, fruited and sporulated commonly in current‐year cone scales of P. abies, P. omorika and P. glauca, while P. rubens, P. mariana and P. pungens appeared to be resistant during the study period. Chrysomyxa ledi did not affect seed development in infected cones. Uredinia of T. areolata frequently occurred on leaves of 41 Finnish and Russian cultivars, varieties or subspecies of Prunus padus L. ssp. badus and ssp. borealis and Pr. virginiana both in the botanical garden and in the field, while 13 exotic Prunus spp. lacked rust fruitbodies. All the Pr. padus cultivars were highly susceptible to T. areolata, thus, spreading the rust efficiently to surroundings. This is the first report of aecia of T. areolata in cones of P. engelmannii and P. glauca, and those of C. ledi in cones of P. omorika and P. glauca. Molecular identification confirmed the presence of T. areolata and C. pirolata on all hosts, and all samples of C. ledi belonged to the C. ledi‐rhododendri complex.     

Germination capacity of Thekopsora areolata aeciospores and the effect of cone rusts on seeds of Picea abies

Mature Norway spruce (Picea abies) cones of different ages and colonized by Thekopsora areolata were collected in northern Finland 2009–2010. The germination capacity of aeciospores collected from intact aecia was measured on water agar and compared across cone age. Seed formation was investigated in healthy cones and those infected by T. areolata, Chrysomyxa pirolata, Chrysomyxa ledi, and seed germination was also measured for healthy cones and those infected by T. areolata. Unsporulated aecia of T. areolata occurred in 0- to 4-year-old cones. Aeciospores from current-year cones did not germinate and the germination capacity was significantly higher in one- to three-year-old cones than in older or current-year cones. However, a small number of aeciospores remained viable in four-year-old cones. Seed number and apparent health were significantly lower in cones infected by T. areolata and C. pirolata than in healthy cones or cones infected by C. ledi. Seed germination was significantly lower in cones infected with T. areolata compared to seeds from healthy cones. Our results suggest that infected cones more than one-year old continue to spread T. areolata for several years after the initial infection and fruiting, and thus promote the likelihood of epidemics and virulence of local outbreaks. Although cones infected by T. areolata and C. pirolata form significantly fewer seeds and of diminished viability, C. ledi does not appear to affect seed formation and health in Norway spruce. We recommend all rust-infected cones to be removed from orchards to improve stand health, reduce the risk of local outbreaks, and thereby increase the production and quality of Norway spruce seed.     

Development of microsatellite markers for Thekopsora areolata,the causal agent of cherry spruce rust

Cherry spruce rust is a fungal disease of Norway spruce cones caused by Thekopsora areolata and responsible for significant losses in seed production in Sweden and Finland. Here, we report the first set of nine microsatellites, which will allow an effective genetic fingerprinting of T. areolata. The markers were isolated using the FIASCO method and were characterized using DNA from 49 single aecia sampled from spruce cones in three different seed orchards in Sweden. Eight of the nine markers were shown to be polymorphic among the aecia. The markers were unlinked and are therefore suitable for future population genetic studies.     

Susceptibility of Ribes spp. to pine stem rusts in Finland

Susceptibility of Ribes spp. to three pine stem rusts, Cronartium ribicola, Cronartium flaccidum and Peridermium pini, was investigated by inoculations both in laboratory and in greenhouse conditions, and by observing sporulation on Ribes spp. leaves under natural and artificial inoculum in 16 experiments. Twenty‐seven Ribes spp. cultivars were inoculated in 2000–2004 using 41 sources of C. ribicola aeciospores from a wide geographic range, and six Pinus spp.; 51 sources of C. flaccidum and 11 sources of P. pini from Pinus sylvestris. The results were very similar both after artificial inoculations and observations under natural inoculum in repeated experiments over the years. Cronartium ribicola uredinia and telia developed frequently or moderately on nine Ribes nigrum cultivars but were not present on two cultivars. Sporulation developed on five Ribes rubrum cultivars but was absent on two cultivars. Three Ribes uva‐crispa cultivars were only weakly susceptible to C. ribicola, while all R. alpinum cultivars were resistant to the rust. The other Ribes spp. tested, Ribes niveum, Ribes aureum, Ribes odoratum, Ribes sp. × nigrolaria, Ribes glandulosum and Ribes × culverwellii Jostaberry, were all susceptible to C. ribicola. Cultivars of R. nigrum were more susceptible than those of any other species. No significant variation in virulence of the aeciospores was observed between and among Ribes hosts over the years. Cronartium flaccidum and Peridermium pini did not form any uredinia or telia on Ribes spp. in any of the experiments.     

Allozyme frequencies,outcrossing rate and pollen contamination in Picea abies seed orchards

Allozymes of Swedish seed orchard clones of Norway spruce (Picea abies Karst.) were studied. There were differences between gene frequencies of origins from different regions. The gene pool of the fertilizating pollen of two different orchards was different. Pollen contamination and outcrossing rates were estimated in two seed orchards. The fraction of paternal gametes which did not originate from any seed orchard clone was 10% and 17%, respectively. Based on simulation it was estimated that only a quarter of the gametes with origin outside the orchard would be detected. Estimation of multilocus outcrossing rates were 0.95 and 0.98.     

Cronartium flaccidum produces uredinia and telia on Melampyrum nemorosum and on Finnish Vincetoxicum hirundinaria

Uredinia and telia were produced on Melampyrum nemorosum through inoculations with Cronartium flaccidum aeciospores collected from northern Finland, proving that M. nemorosum serves as the telial host of C. flaccidum. Melampyrum nemorosum plants were either grown from seeds collected from several geographical provenances or transplanted from natural forest stands. They were inoculated with several geographical aeciospore sources of C. flaccidum in a greenhouse. Vincetoxicum hirundinaria, M. pratense and M. sylvaticum were used as positive controls. Detached leaves of Melampyrum spp. and V. hirundinaria were also inoculated but uredinia and telia formed only on V. hirundinaria. Most of the aeciospore sources from northern Finland produced abundant uredinia and telia both on M. nemorosum from two geographical locations and on V. hirundinaria in the greenhouse. Statistically significant differences were observed in the dimensions of artificially produced telia, teliospores and urediniospores among the tested alternate hosts and spore sources. The results suggest that M. nemorosum is susceptible to C. flaccidum and therefore may play a role in southern Fennoscandia as an alternate host but it has potentially a greater role in central and eastern Europe, where M. nemorosum occurs commonly.     

Pathogenicity of Ceratocystis resinifera to Norway spruce

The blue‐stain fungus Ceratocystis resinifera colonizes wounds on living Picea spp. and other conifers in Europe and North America. Little is known regarding the pathogenicity of this fungus and consequently, four Norwegian C. resinifera isolates were inoculated on to Norway spruce (Picea abies) using two different techniques. These included single‐point inoculations on young trees (two inoculations per tree on 14‐year‐old trees) and mass‐inoculations on older trees (～200 inoculations per tree on 34‐year‐old trees). In both experiments, C. resinifera induced minor symptoms that in most cases did not differ significantly from inoculation with sterile agar. The virulent blue‐stain fungus C. polonica, which was inoculated for comparative purposes, induced extensive symptoms, causing 83% dead cambium circumference and 82% blue‐stained sapwood, and long necrotic lesions in the phloem. The results suggest that C. resinifera is non‐pathogenic or only mildly pathogenic to Norway spruce and does not present a threat to these trees.     

Chrysomyxa ledi,a new rust fungus sporulating in cone scales of Picea abies in Finland

Abstract

Seasonal fruiting and sporulation of cone rusts were investigated in Norway spruce cones during the growing season of 2007, a year after an excellent cone crop and a severe outbreak of rust. Current-year and 1-year-old cones were collected bimonthly to monthly from case-stands in southern and northern Finland and checked for rust fruitbodies. A previously unreported species of rust formed spermogonia and aecia, and sporulated on the cone scales of Norway spruce in two study areas. Morphologically, the rust was distinct from other spruce cone rusts and, based on ultrastructure of aecia and aeciospores, most closely resembled Chrysomyxa ledi. Molecular analysis of the aeciospores confirmed that the rust belonged to the C. ledi/C. rhododendri complex. The rust fruited frequently but only colonized a few scales per cone and presumably had a minor effect on cone development and seed crop.     

Life cycle and taxonomy of Chrysomyxa succinea in China and phylogenetic positions of Caeoma species on Rhododendron

Rust diseases of spruce and rhododendrons caused by Chrysomyxa succinea are commonly observed in forests of northern and eastern Asia. The heteroecious life cycle of this species alternating between spermogonial and aecial stages on Picea jezoensis and uredinial and telial stages on Rhododendron aureum is confirmed for the first time in China by inoculations and phylogenetic analyses. Morphological characters of all stages are described based on specimens obtained from the field and from inoculations. Based on morphological similarities and phylogenetic analyses, Ch. zhouniensis reported on Picea asperata is treated as a synonym of Ch. succinea. Four Caeoma species on Rhododendron are included in the same phylogenetic group as species of Chrysomyxa. Therefore, these species are treated as Chrysomyxa, and four new combinations in Chrysomyxa are proposed.     

Seedborne fungi in Norway spruce: testing methods and pathogen control by seed dressing

Italian seed lots of Norway spruce (Picea abies) were analysed by agar plate tests and seedling symptom test. Fusarium spp., Phoma spp. and Sirococcus conigenus were present on seeds, together with some saprophytes, but only S. conigenus and Fusarium spp. caused mortality in the seedling symptom test. Pathogenicity tests with 19 isolates belonging to nine Fusarium species showed that only some were pathogenic. Fusarium dlamini, reported for the first time in Italy, caused only some pre-emergence mortality. Pathogenicity tests with S. conigenus indicated that the isolate was pathogenic and that temperature can greatly affect the results. Seed dressing with prochloraz was suitable for controlling the seedborne pathogens. Agar plate tests are effective and practicable methods for the detection of spruce seed mycoflora.     

New Picea hosts for Chrysomyxa ledi and Thekopsora areolata

A severe epidemic of Chrysomyxa ledi needle rust occurred over large areas in northern Finland in 2015. In late summer, young current‐year cones and needles of 7–8 Picea species were sampled in a northern botanical garden, and the frequencies of cone rusts, Thekopsora areolata, Chrysomyxa pirolata and C. ledi, were investigated in cones, and C. ledi was investigated in needles. In addition, the frequency of C. ledi was estimated visually in current‐year needles of ca. 400 trees of 17 species and varieties of Picea in the garden. Chrysomyxa pirolata was absent in cones of all Picea. No cone rusts were observed in cones of P. rubens and Picea mariana. Aecia of Thekopsora areolata occurred moderately in cones of P. abies (26% of cones) and rarely in P. engelmannii (3%) and P. omorika (6%). Chrysomyxa ledi was the most frequent rust in cones; aecia occurred abundantly in cones of P. abies (100%), P. engelmannii (69%), P. glauca (45%) and P. omorika (41%), and occasionally on P. pungens (13%). Thekopsora areolata from cones of the three Picea species showed 100% similarity in ITS2 sequence to one another and to samples in the GenBank. Samples of Chrysomyxa from needles and cones of Picea and leaves of Ledum palustre had ITS2 sequences 99.9%–100% similar to one another and 99.8%–100% similar to the closest sequence of C. ledi and C. rhododendri in the GenBank. Thekopsora areolata was recorded for the first time in cones of P. omorika and C. ledi in cones of P. engelmannii and P. pungens. In current‐year needles of Picea, C. ledi was abundant on P. abies, P. obovata, P. engelmannii, P. jezoensis, P. glauca and P. sitchensis, the rust was scarce on P. asperata var. nobilis, P. mariana, P. pungens and P. omorika, and the rust was absent on P. glehnii and P. rubens.     

Rhizomorph production and stump colonization by co‐occurring Armillaria cepistipes and Armillaria ostoyae: an experimental study

In managed spruce forests, Armillaria cepistipes and A. ostoyae are efficient stump colonizers and may compete for these resources when they co‐occur at the same site. The aim of this experiment was to quantify the mutual competitive ability of the two Armillaria species in producing rhizomorphs and in colonizing Norway spruce (Picea abies) stumps. Five isolates of A. cepistipes and two isolates of A. ostoyae were simultaneously inoculated pair‐wise into pots containing a 4‐year‐old spruce seedling. For comparison, each isolate was also inoculated alone. One year after inoculation, stumps were created by cutting down the seedlings. Six months after creation of the stumps, rhizomorph production and stump colonization were assessed. Armillaria spp. were identified from 347 rhizomorphs and 48 colonized stumps. Armillaria cepistipes dominated both as rhizomorphs in the soil and on the stumps. Nevertheless, A. ostoyae was relatively more frequent on the stumps than in the soil and A. cepistipes was relatively more frequent in the soil than on the stumps. In both species, the ability to colonize the stumps in simultaneous inoculations was significantly reduced compared with single inoculations. In respect to rhizomorph production, simultaneous co‐inoculations had a slightly stimulatory effect on A. cepistipes and no significant effect on A. ostoyae. Our study suggests a rather neutralistic co‐existence of A. cepistipes and A. ostoyae as rhizomorphs in the soil. Concerning the ability to colonize stumps, the two species experience a mutual negative effect from the interaction, probably because of interspecific competition.     

Pathogenicity of Phytophthora species and Pythium undulatum isolated from Abies procera Christmas trees in Ireland

Phytophthora cryptogea, Phytophthora cinnamomi, Phytophthora cambivora, Phytophthora megasperma and Pythium undulatum were isolated from diseased Noble fir (Abies procera) seedlings and soil associated with dead Noble fir in Ireland. Seedlings of four Christmas tree species (A. procera, Picea sitchensis, Picea abies and Pinus contorta) were inoculated with these oomycetes to test their pathogenicity and the susceptibility of the various tree species. Phytophthora spp. and Pythium undulatum caused root rot on all tree species. Disease symptoms included reddish brown cambial discoloration, crown symptoms, brown foliage, dark brown roots, root rot and seedling mortality. These symptoms were similar to those observed on Noble fir in naturally infested plantations. Pythium undulatum appeared as the most virulent pathogen followed by P. cinnamomi, P. cambivora, P. megasperma and P. cryptogea. Noble fir showed to be most susceptible and lodgepole pine most tolerant while Sitka spruce and Norway spruce were intermediate.     

Improved varieties perform well in realized genetic gain trials with Norway spruce seed sources in southern Sweden

ABSTRACT

Genetically improved Norway spruce (Picea abies L. Karst) has been extensively used in operational plantations in Sweden. However, there have been few studies that aimed at verifying the realized genetic gain over a longer time span in plots established with one genetical entry. Two experimental series at six sites in southern Sweden, established purely for this purpose, were assessed. Early height growth of 10 Norway spruce seedlots of different genetic levels was compared. There were significant differences in height at the age of 5 and 8 years between improved and unimproved seed sources and the differences between most advanced material from the breeding front, represented by full-sib crossings of selected plus-trees, and unimproved Swedish spruce were 36% and 30% at the age of 5 and 8 years, respectively. At the same ages, the trees originating from the second round of seed-orchards were 25% and 21% higher than unimproved material. There were also about 20% differences between the best and worst tested full-sib families, indicating large possible gain in the new generation. The study confirmed earlier estimates of genetic gain levels of different seed sources in the breeding program for Norway spruce.     

Pathogenicity of ophiostomatoid fungi on Picea abies in Slovenia

Pathogenic fungi can survive and develop in living plants, often causing diseases in the host. Some theories speculate that pathogenic ophiostomatoid fungi provide benefits to its vectors – bark beetles – by overcoming the tree's defence mechanisms. This study reports the results of an experiment in south‐eastern Europe in which mature and seedling Norway spruce trees were artificially inoculated with various ophiostomatoid fungi. The aim of the experiment was to determine the relative virulence of ophiostomatoid fungi by assessing the ability of the fungi to stimulate host tree defence mechanisms through inoculation experiments. Experiments were performed by inoculation of Picea abies in seedling and mature trees. The following fungi were used in low‐density and seedling inoculations: Ophiostoma ainoae, O. brunneo‐ciliatum, Grosmannia cucullata and an unidentified Leptographium sp., O. bicolor, O. fuscum, O. piceae, G. penicillata and G. piceiperda. Endoconidiophora polonica was used in mass and seedling inoculations. Various characteristics such as host vitality, blue stain, lesion and resin outflow were measured before and after the trees were felled. E. polonica caused blue stain, induced large lesions and killed some of the mature trees and seedlings, confirming earlier reports that it is a strong wound pathogen. Only E. polonica, Leptographium sp. and O. ainoae caused blue stains in the sapwood of inoculated seedlings. In low‐density inoculations, G. piceiperda induced intense necrosis and had higher values for all the characteristics monitored. Some of the other ophiostomatoid fungi showed a moderate level of pathogenicity. Fungi with the capacity to stimulate a host defence mechanism could play a role in the establishment of bark beetle populations.     

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.     

Potential Effects of Temperature on Early Reproductive Development and Progeny Performance in Picea abies (L.) Karst

In order to enhance cone production in Norway spruce [Picea abies (L.) Karst] seed orchards have been established in more southerly, warmer sites in Norway. This has led to concern and some evidence that seedlings obtained from parent trees grown at warmer sites may not be well adapted if planted in northern regions. Selective events during sexual reproduction in the warmer climate may select for traits not well adapted for cold climates. This study describes ovule and male gametophyte development, fertilization and proembryo and early embryo development under six different temperature regimens of warm or cold, warm-cold or cold-warm, and compares development in relation to temperature sums in degree days (dd) and calendar date. The outside treatment in a cold climate in Norway served as the control, and normal development and few ovule abortions and abnormalities were observed. In no treatments were abnormalities in male gametophyte development observed. In treatments in which trees were kept in the greenhouse throughout the study or moved during the study from the outside to inside the greenhouse, a higher incidence of megagametophyte, egg or embryo abortion was generally observed.     

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

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

Susceptibility of Pedicularis spp. to Cronartium ribicola and C. flaccidum in Finland

Aeciospores of Cronartium ribicola and C. flaccidum were collected from several locations in Finland and used to inoculate Pedicularis spp. and some known and suspected alternate hosts in 2008–2009. In all trials, C. ribicola formed uredinia and telia on leaves of Ribes nigrum. No uredinia or telia of C. ribicola formed on older leaves of Pedicularis spp. but both uredinia and telia were found on young leaves of P. palustris ssp. palustris. Cronartium flaccidum produced uredinia and/or telia on leaves of P. palustris ssp. palustris, P. lapponica, Vincetoxicum hirundinaria and Melampyrum sylvaticum. Neither rust infected P. sceptrum‐carolinum, Vaccinium myrtillus, Calluna vulgaris or Ledum palustre. Similar to rusts in Asia and North America, the results showed that European C. ribicola exhibit more variable host reactions and wider alternate host ranges than earlier described. Pedicularis palustris may play a role in the spread of Cronartium in natural forests.     

Comparison of colonization capacity by asexual spores of Heterobasidion species in Norway spruce wood

Two species of the tree pathogenic fungus Heterobasidion spp. exist in Sweden, Heterobasidion annosum s.s. and Heterobasidion parviporum. Both species are known to infect Norway spruce (Picea abies). The aim of the study was to examine the interspecific competition between H. annosum s.s. and H. parviporum as well as their colonization rate in fresh Norway spruce wood. Equal amount of conidiospores from each species was sprayed together on 30 fresh, previously uninfected, Norway spruce billets. After incubation in a greenhouse, the proportion of Heterobasidion spp. colonies belonging to each species was recorded. Of the 196 colonies isolated from the upper part of billets, 195 were H. parviporum. All isolated colonies further down in the billets were H. parviporum. To study the colonization rate, H. annosum s.s. and H. parviporum were sprayed alone on 30 spruce billets each, incubated and growth recorded both vertically and horizontally. H. parviporum grew further down in the billets (p = 0.008) and covered a larger area (p < 0.001) than H. annosum s.s. While H. annosum s.s. and H. parviporum both infected fresh Norway spruce wood H. parviporum outgrew and outcompeted H. annosum s.s during the early colonization stage.