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1.
Root-associated fungal communities from two phenologically contrasting Silver Fir (Abies alba Mill.) groups of trees
Tina Unuk, Tijana Martinović, Domen Finžgar, Nataša Šibanc, Tine Grebenc, Hojka Kraigher, 2019

Abstract: Root-associated fungal communities are important components in ecosystem processes, impacting plant growth and vigor by influencing the quality, direction, and flow of nutrients and water between plants and fungi. Linkages of plant phenological characteristics with belowground root-associated fungal communities have rarely been investigated, and thus our aim was to search for an interplay between contrasting phenology of host ectomycorrhizal trees from the same location and root-associated fungal communities (ectomycorrhizal, endophytic, saprotrophic and pathogenic rootassociated fungi) in young and in adult silver fir trees. The study was performed in a managed silver fir forest site. Twenty-four soil samples collected under two phenologically contrasting silver fir groups were analyzed for differences in rootassociated fungal communities using Illumina sequencing of a total root-associated fungal community. Significant differences in beta diversity and in mean alpha diversity were confirmed for overall community of ectomycorrhizal root-associated fungi, whereas for ecologically different non-ectomycorrhizal root-associated fungal communities the differences were significant only for beta diversity and not for mean alpha diversity. At genus level root-associated fungal communities differed significantly between early and late flushing young and adult silver fir trees. We discuss the interactions through which the phenology of host plants either drives or is driven by the root-associated fungal communities in conditions of a sustainably co-naturally managed silver fir forest.
Keywords: host phenology, stand age, root-associated fungi, silver fir, fungal community
DiRROS - Published: 20.02.2020; Views: 1317; Downloads: 980
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2.
3.
Ectomycorrhizal community composition of organic and mineral soil horizons in silver fir (Abies alba Mill.) stands
Tanja Mrak, Emira Hukić, Ines Štraus, Tina Unuk, Hojka Kraigher, 2020

Abstract: Vertical ectomycorrhizal (ECM) community composition was assessed on silver fir (Abies alba Mill.) in beech-silver fir forests in Bosnia and Herzegovina. Organic and upper mineral horizons were described by pedological analyses. Silver fir root tips were divided into vital ECM, old and non-mycorrhizal for each horizon separately. Morpho-anatomical classification of vital ECM root tips with an assessment of abundance was followed by ITS-based molecular characterization and classification into exploration types. The percentage of vital ECM root tips was not affected by the soil horizon. Altogether, 40 ECM taxa were recorded. Several taxa have not previously been reported for silver fir: Hebeloma laterinum, Inocybe fuscidula, I. glabripes, Lactarius acris, L. albocarneus, L. blennius, L. fluens, Ramaria bataillei, Russula badia, R. lutea, R. mairei, Sistotrema sp., Tarzetta catinus, Tomentella atroarenicolor, T. badia, T. cinerascens, T. bryophylla, and T. ramosissima, indicating high potential for diversity of ECM fungi in silver fir stands. No significant differences in community composition, species richness and diversity were detected between mineral and organic horizons. Community composition was affected by CaCO3, organic carbon, organic carbon stock, total nitrogen stock, C:N ratio and soil density. No significant effects of soil parameters were detected for exploration types. The contact exploration type was dominant in both soil horizons. Significantly different relative abundances of dominant taxa Tomentella stuposa, Cenococcum geophilum and Piloderma sp. 1 were detected in the two horizons. Twelve taxa were limited to the organic horizon and eight to the mineral horizon.
Keywords: biodiversity, ectomycorrhizal fungi (ECM), beech-silver fir forest, ECM depth profile, Balkan 30 Peninsula, ectomycorrhizal exploration types
DiRROS - Published: 17.06.2020; Views: 1330; Downloads: 435
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4.
PCR primers comparisons for a successful Tuber spp. DNA region amplification in routine identifications
Tina Unuk, Hojka Kraigher, Tine Grebenc, 2020

Abstract: Since late 20th century DNA sequencing became the method of choice method in precision species identification. The ITS region is one of the official fungal barcoding DNA markers, although in some cases sequencing of the ITS re-gion may, due to misidentification, mislabeling or nomen-clature errors in public databases, lead to incorrect or insuf-ficient identification, as is currently a case in the genus Tu b e r. The aim of this study was to test, which ITS primer pairs are most appropriate and optimal for Tu b e r species DNA region amplification. Thereby we (1) compared ampli-fication success for different Tu b e r species using fungal spe-cific primer pair ITS1f and ITS4 and (2) compared amplifi-cation success using different ITS primer pair combinations in amplifying DNA region an example species Tuber aesti-vum. Based on results, Tuber aestivum was one of the most reluctant Tu b e r species in this study and in most cases failed to amplify with the above primer pair. After comparing dif-ferent ITS primer pairs, we conclude that the primer pair ITS5 and ITS7 is the most appropriate primer pair for ampli-fication DNA region of T. ae stiv um as it resulted in high am-plification success from ectomycorrhizal root tips. Based on sequences, gained from public databases, we found that ITS1f and ITS6 primers have a mismatch in one base pair compared to the target sequence of Tuber aestivum, thus re-sulting in poor or no amplification success. Although prim-er pair ITS5 and ITS7 in our study was proven to be the most appropriate primer pair in amplifying DNA region Tu b e r aestivum species, further analysis about appropriateness of it for a general barcoding and identification of ectomycorrhiza in complex community samples is needed.
Keywords: Tuber spp., ITS region, PCR amplification, ITS primers
DiRROS - Published: 30.07.2020; Views: 984; Downloads: 727
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5.
Effect of earthworms on mycorrhization, root morphology and biomass of silver fir seedlings inoculated with black summer truffle (Tuber aestivum Vittad.)
Tina Unuk, Niccolo G. M. Benucci, Tine Grebenc, Hojka Kraigher, 2021

Abstract: Species of the genus Tuber have gained a lot of attention in recent decades due to their aromatic hypogenous fruitbodies, which can bring high prices on the market. The tendency in truffle production is to infect oak, hazel, beech, etc. in greenhouse conditions. We aimed to show whether silver fir (Abies alba Mill.) can be an appropriate host partner for commercial mycorrhization with truffles, and how earthworms in the inoculation substrate would affect the mycorrhization dynamics. Silver fir seedlings inoculated with Tuber. aestivum were analyzed for root system parameters and mycorrhization, how earthworms affect the bare root system, and if mycorrhization parameters change when earthworms are added to the inoculation substrate. Seedlings were analyzed 6 and 12 months after spore inoculation. Mycorrhization with or without earthworms revealed contrasting effects on fine root biomass and morphology of silver fir seedlings. Only a few of the assessed fine root parameters showed statistically significant response, namely higher fine root biomass and fine root tip density in inoculated seedlings without earthworms 6 months after inoculation, lower fine root tip density when earthworms were added, the specific root tip density increased in inoculated seedlings without earthworms 12 months after inoculation, and general negative effect of earthworm on branching density. Silver fir was confirmed as a suitable host partner for commercial mycorrhization with truffles, with 6% and 35% mycorrhization 6 months after inoculation and between 36% and 55% mycorrhization 12 months after inoculation. The effect of earthworms on mycorrhization of silver fir with Tuber aestivum was positive only after 6 months of mycorrhization, while this effect disappeared and turned insignificantly negative after 12 months due to the secondary effect of grazing on ectomycorrhizal root tips.
Keywords: mycorrhiza, truffles, silver fir, inoculation, earthworms, spore inoculation
DiRROS - Published: 22.03.2021; Views: 504; Downloads: 325
.pdf Fulltext (1,32 MB)

6.
Global homogenization of the structure and function in the soil microbiome of urban greenspaces
Manuel Delgado-Baquerizo, David J. Eldridge, Yu-Rong Liu, Blessing Sokoya, Jun-Tao Wang, Hang-Wei Hu, Ji-Zheng He, Felipe Bastida, José L. Moreno, Adebola R. Bamigboye, Tine Grebenc, Tina Unuk, 2021

Abstract: The structure and function of the soil microbiome of urban greenspaces remain largely undetermined. We conducted a global field survey in urban greenspaces and neighboring natural ecosystems across 56 cities from six continents, and found that urban soils are important hotspots for soil bacterial, protist and functional gene diversity, but support highly homogenized microbial communities worldwide. Urban greenspaces had a greater proportion of fast-growing bacteria, algae, amoebae, and fungal pathogens, but a lower proportion of ectomycorrhizal fungi than natural ecosystems. These urban ecosystems also showed higher proportions of genes associated with human pathogens, greenhouse gas emissions, faster nutrient cycling, and more intense abiotic stress than natural environments. City affluence, management practices, and climate were fundamental drivers of urban soil communities. Our work paves the way toward a more comprehensive global-scale perspective on urban greenspaces, which is integral to managing the health of these ecosystems and the well-being of human populations.
Keywords: soil biodiversity, structural diversity, functional diversity, urban soils
DiRROS - Published: 15.07.2021; Views: 423; Downloads: 282
.pdf Fulltext (4,34 MB)

7.
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Potential link between ectomycorrhizal community composition and host tree phenology
Tina Unuk, Rok Damjanić, Hojka Kraigher, Tine Grebenc, 2021

Abstract: The timing of leaf phenology tends to be crucial in controlling ecosystem processes such as the acquisition of carbon and water loss as well as in controlling tree nutrient cycling. To date, tree phenology has mostly been associated with environmental control (e.g., temperature and photoperiod) in a relationship with inheritance, while it has rarely been linked with ectomycorrhizal community compositional changes through the host tree’s phenological stages. Seasonal variations of fungal communities have been widely studied, but little is known about mycorrhiza community composition changes along phenological stages. Therefore, we analyzed ectomycorrhizal communities associated with silver fir and their compositional changes during the transition between phenological stages. The phenological stages of each individual tree and time of bud break were monitored weekly for two years and, at the same time, ectomycorrhiza was harvested from selected silver fir trees. In total, 60 soil cores were analyzed for differences in the ectomycorrhizal community between phenological stages using Sanger sequencing of individual ectomycorrhizal morphotypes. A significant difference in beta diversity for an overall ectomycorrhizal community was confirmed between analyzed time periods for both sampled years. Species-specific reactions to transitions between phenological stages were confirmed for 18 different ectomycorrhizal taxa, where a positive correlation of Russula ochroleuca, Russula illota, Tomentella sublilacina, and Tylospora fibrillosa was observed with the phenological stage of bud burst.
Keywords: Abies alba (Mill.), ectomycorrhizal community, phenological stages
DiRROS - Published: 20.12.2021; Views: 222; Downloads: 118
.pdf Fulltext (1,73 MB)

9.
Buckwheat milling waste effects on root morphology and mycorrhization of Silver fir seedlings inoculated with Black Summer Truffle (Tuber aestivum Vittad.)
Tina Unuk, Tine Grebenc, Daniel Žlindra, Tanja Mrak, Matevž Likar, Hojka Kraigher, Zlata Luthar, 2022

Abstract: Large amounts of buckwheat waste are generated annually by the industry and are used in several different ways. To date, there has been little research regarding its suitability as a medium for growing seedlings in nurseries. The aim of this study was therefore to analyze the suitability of common and Tartary buckwheat wastes (brans and husks) as media used for raising seedlings. A pot experiment with five different treatments was carried out, in which silver fir root parameters were analyzed and compared 6 and 12 months after summer truffle-spore inoculation. A significantly higher concentration of the antioxidant rutin was confirmed in Tartary buckwheat bran compared to other buckwheat waste used. We also confirmed a significantly positive effect of added Tartary buckwheat husks on specific root length, root tip density, and specific root tip density compared to added common buckwheat husks or Tartary buckwheat bran, for which a significantly negative effect on branching density was confirmed. A significantly negative effect of added buckwheat husks and Tartary buckwheat bran was confirmed for summer truffle mycorrhization level.
Keywords: buckwheat waste, root growth, summer truffle, forest nursery, silver fir, inoculation with ectomycorrhizal fungi
DiRROS - Published: 09.02.2022; Views: 165; Downloads: 132
.pdf Fulltext (1,37 MB)
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