Foliar nematode,Litylenchus crenatae ssp. mccannii,population dynamics in leaves and buds of beech leaf disease‐affected trees in Canada and the US

A foliar nematode, Litylenchus crenatae ssp. mccannii, is associated with beech leaf disease (BLD) symptoms. Information about the types of tissues parasitized and how nematode populations fluctuate in these tissues over time is needed to improve surveys as well as understand the nematodes role in BLD. During this study, the nematode was detected throughout the known range of BLD by researchers at both Canadian and US institutions using a modified pan method to extract nematodes. Monthly collections of symptomatic and asymptomatic leaves during the growing season (May–October), and leaves and buds between growing seasons (November–March), revealed that nematodes were present in all tissue types. Progressively larger numbers of nematodes were detected in symptomatic leaves from Ohio and Ontario, with the greatest detections at the end of the growing season. Smaller numbers of nematodes were detected in asymptomatic leaves from BLD‐infected trees, typically at the end of the growing season. The nematode was detected overwintering in buds and detached leaves. The discovery of small numbers of nematodes in detached leaves at one location before BLD was detected indicates that nematodes may have been present before disease symptoms were expressed. Other nematodes, Plectus and Aphelenchoides spp., were infrequently detected in small numbers. Our findings support the involvement of the nematode in BLD and indicate that symptoms develop only when certain requirements, such as infection of buds, are met. We also found that the nematode can be reliably detected in buds and leaves using the modified pan extraction method.     

<Emphasis Type="Italic">Samardala</Emphasis>: specificities and changes in the ethnobotanical knowledge about <Emphasis Type="Italic">Allium siculum</Emphasis> subsp<Emphasis Type="Italic">. dioscoridis</Emphasis> (Sm.) K. Richt. in Bulgaria

Allium siculum Ucria is a perennial bulbous plant that occurs in Bulgaria as A. siculum subsp. dioscoridis (Sm.) K. Richt. The plant is locally known as “samardala”. Its herbage is used traditionally by local communities in Bulgaria as edible green and for preparation of salty spice mixtures. The study was focused on the documentation of traditional knowledge about samardala, covering local collection and cultivation practices, processing methods and consumption preferences. The respondents (aged 30–86 years, 50% over 65) were local producers and small-scale salesmen of the herbage and samardala-containing spices. To assess the health features of samardala spices the total phenol and flavonoid content was measured in home-consumed or marketed flavoured salts. Overall appreciation of the plant and related food products was found to be very high; presenting a considerable engagement of the locals in plant cultivation and gathering, laborious processing and consumption. Traditional ecological knowledge about the biology, phenology and ecology of the samardala plant and methods for its processing is still preserved by elderly people who rely mostly on their own cultivated plants. Although the introduction of modern appliances has altered the method of production, its main stages and the recipes are preserved. Modernization of the processing method has not affected the quality of the products and faster processing could be a contributing factor for the higher content of biologically active substances. Phenolic content was about twice as low as the flavonoid content, following the same pattern for all tested flavoured salts. Measures for better management of natural populations and promotion of cultivation practices of A. siculum subsp. dioscoridis would ensure the sustainable quality of traditional products as well as conservation of the natural plant resources.     

Forest farming of wine-cap <Emphasis Type="Italic">Stropharia</Emphasis> mushrooms

Cultivation of the wine-cap Stropharia mushroom (Stropharia rugoso-annulata Farlow ex. Murrill) has potential as a forest farming practice in agroforestry. Two outdoor cultivation experiments were established in central Missouri during 2006. We examined Stropharia production in response to two substrate/casing systems: (1) an uncased mixture of lightly composted poplar wood chips and soil, without casing; and (2) leached and partially fermented wheat straw cased with the same chip/soil mixture. We also examined the effect of soil contact (presence/absence of water-permeable fabric), and inoculation season (mid versus late summer) over two field seasons. Mushroom production during both seasons (2006 and 2007) was greater from the straw/chips system than from the wood chip system. The use of ground-cover fabric did not enhance mushroom production. Further research is needed to determine the most productive dimensions of cultivation beds, to evaluate alternative casing methods, and to explore the possibility of cultivation bed renewal with fresh substrate.     

Dispersal of Contaminant Metals in the Mining-Affected Danube and Maritsa Drainage Basins, Bulgaria, Eastern Europe

Metal dispersal in the Danube and Maritsa drainage basins resulting from metal mining activities in Bulgaria has been assessed through the collection of 611 samples of river water, river channel and floodplain sediment, and mine waste from over 218 sites. Concentrations of Cd, Cu, Pb, and Zn in river water were found to be highest in close proximity to locations of Cu and Pb–Zn mining regions in the Maritsa catchment. Downstream dispersal of solute metals in these catchments, and into the River Danube, was found to be limited by physical dilution and a well-buffered pH environment. Dispersal of contaminant metals in channel and floodplain sediment was found to be extensive. Contamination was particularly severe in the Rivers Timok and Iskar (Danube catchment) and the Topolnitsa, Chepelarska, and Arda Rivers (Maritsa catchment) and creates the potential of transboundary dispersal of contaminant metals.     

An improved in vitro technique for isolated microspore culture of barley

A detailed procedure for isolated microspore culture of barley is presented along with examples of response across genotypes. Over 30 genotypes, including winter and spring growth habit and 2-row and 6-row genotypes, have shown an essentially genotype independent response, averaging about 10,000 embryos per 5 cm petri culture plate. The regeneration frequency, checked on samples of 500 embryos per plate ranged from 36 to 97% with most genotypes being in the range of 70 to 90%. About 70 to 80% of the plants regenerated have been completely fertile doubled haploids, thus eliminating the need to double the chromosome number of plants. Many little details are critical to success of the microspore procedure and while it saves much time compared to anther culture, greater attention to details and cleanliness is essential. This revised version was published online in August 2006 with corrections to the Cover Date.     

Truffle seedling production method has long-term consequences for tree growth and root colonization

Cultivation of Burgundy black truffles (Tuber aestivum syn. T. uncinatum) in Midwestern agroforestry has the potential to provide important income not only to landowners, but also to the hospitality industry. Economically viable harvest depends upon both successful seedling colonization by the truffle fungus and successful growth of the fungus along with the extending root system. We evaluated an established hybrid oak (Quercus bicolor × Q. robur) truffière 5 and 6 years after planting in May 2005, to evaluate the effects of three seedling production methods on tree growth and root colonization by T. aestivum. Oak seedlings produced using two variations on the RPM^© (Root Production Method) substrate grew significantly faster in height and diameter, but were significantly less well colonized by the truffle fungus compared with saplings from seedlings produced by the “Typical” method. We found that 0.7–19.8 % of root tips (mean 5.1 %) sampled from saplings grown from Typical seedlings were colonized by T. aestivum. By 2010, two distinct canopy forms had developed: ‘upright’ versus ‘spreading’. Although root colonization was not related to canopy form, future fruit body production may be influenced by the greater soil shading provided by the spreading canopy form. A comparison of autumn/winter air temperatures in Missouri, USA with European conditions suggests that fruit body production in Missouri will likely be greatest from mid-September through mid-December. Soil pH modification by application of crushed limestone prior to planting was effective in maintaining average pH at 7.16 through 2010, a level consistent with fruit body production.     

Colonization of Pedunculate oak by the Burgundy truffle fungus is greater with natural than with pelletized lime

European black truffles can be profitable agroforestry crops outside their native ranges. Truffle fungi grow symbiotically as ectomycorrhizae on the roots of host trees, notably hazels and oaks. Conditions in the central USA appear conducive to cultivation of the Burgundy truffle (Tuber aestivum Vitt. syn. T. uncinatum Chatin), but research is needed to determine effects of management practices on truffle establishment and fruiting. In a greenhouse study we tested the effect of lime type, inoculation technique, and two truffle sources on Pedunculate oak (Quercus robur L.) growth and mycorrhizal colonization. We found that the type of lime used to raise potting mix pH can differentially affect the growth rate of root systems inoculated with different selections of Burgundy truffle inoculum. Seedlings inoculated with one selection of the truffle and grown in potting mixes amended with natural crushed dolomitic limestone developed larger root systems with more truffle mycorrhizae compared with potting mix amended with high-calcium pelletized quick-release lime. Seedlings inoculated with a second truffle selection were not affected by lime source and developed root systems as large as those developed with the first truffle source grown with natural lime. Supplemental root dip inoculation did not improve levels of colonization beyond those accomplished by potting mix infestation with truffle ascospores. Use of a hygroscopic polymer to maintain ascospore suspension in the inoculum slurry used to infest the potting mix had no effect on root system development or mycorrhiza formation.     

Beech leaf disease symptoms caused by newly recognized nematode subspecies Litylenchus crenatae mccannii (Anguinata) described from Fagus grandifolia in North America

Symptoms of beech leaf disease (BLD), first reported in Ohio in 2012, include interveinal greening, thickening and often chlorosis in leaves, canopy thinning and mortality. Nematodes from diseased leaves of American beech (Fagus grandifolia) sent by the Ohio Department of Agriculture to the USDA, Beltsville, MD in autumn 2017 were identified as the first recorded North American population of Litylenchus crenatae (Nematology, 21, 2019, 5), originally described from Japan. This and other populations from Ohio, Pennsylvania and the neighbouring province of Ontario, Canada showed some differences in morphometric averages among females compared to the Japanese population. Ribosomal DNA marker sequences were nearly identical to the population from Japan. A sequence for the COI marker was also generated, although it was not available from the Japanese population. The nematode was not encountered in Fagus crenata (its host in Japan) living among nematode‐infested Fagus grandifolia in the Holden Arboretum, nor has L. crenatae been reported in American beech in Japan. The morphological and host range differences in North American populations are nomenclaturally distinguished as L. crenatae mccannii ssp. n. from the population in Japan. Low‐temperature scanning electron microscopy (LT‐SEM) demonstrated five lip annules and a highly flexible cuticle. Females, juveniles and eggs were imaged within buds with a Hirox Digital microscope and an LT‐SEM. Nematodes swarmed to the tips of freshly cut beech buds, but explants could not be maintained. Inoculation of fresh nematodes from infested leaves or buds to buds or leaves of F. grandifolia seedlings resulted in BLD leaf symptoms. Injuring dormant buds prior to nematode application, in fall or spring, promoted the most reliable symptom expression. The biogeography and physiology of anguinid nematode leaf galling, and potential co‐factors and transmission are discussed.