CULTURE MEDIA AND PROCESS FOR IMPROVED ISOLATION AND MAINTENANCE OF TERFEZIA SPP. MYCELIUM CULTURES

The present invention relates to a culture media and to a process for improved isolation and maintenance of mycelium cultures of “desert truffles” included within the genus Terfezia, the most species rich of all of desert truffle genera. Cultivation of these ectomycorrhizal Ascomycota implies the co-culture of both fungal symbiont and plant host in sterile or semi-sterile conditions. However, isolation and maintenance of Terfezia spp. pure cultures can be challenging. Moreover, many strains are unable to be sub-cultured, and so far, the few successful attempts, mainly with Terfezia spp. from alkaline soils, were found to grow much too slowly to produce adequate amounts of mycelial inoculum in conventional culture media and conditions

Macrofungi of Mata da Margaraça (Portugal), a relic from the Tertiary Age

Mata Nacional da Margaraça represents a rare example where the Atlantic climate influences the perpetuity of a small enclave of a previously widespread laurel forest. The higher relative humidity values (> 80%), which are almost constant all year long and the mild temperatures, rarely exceeding 30°C, even in the dry season (June to September), create an insular-like effect. The biological communities of Margaraça exhibit a transitory character. The forest is dominated by Quercus robur and Castanea sativa, yet Quercus suber, although less frequent, can also be found. The laurel species, such as Viburnum tinus, Ilex aquifolium, Laurus nobilis and the Portuguese endemic Prunus lusitanica ssp. lusitanica, relics from the ancient Portuguese Tertiary, comprise the understorey. The present work represents, to the best of our knowledge, the first regional macrofungal species list of the Mata da Margaraça published to date. The recent fires that occurred in the area have provided the opportunity to study the post-fire communities. The surveys were carried out in 2004 and later in 2018-2019. A total of 272 species were registered as belonging to Basidiomycota (≈ 80%) and Ascomycota (≈ 20%). The most represented Basidiomycota families were Russulaceae, Mycenaceae and Agaricaceae and the most represented Ascomycota families were Pyronemataceae and Pezizaceae. The new records to Portugal add up to a total of 74 species and another 116 species are new records to the province of Beira Litoral. Post-fire fungi account for 17 of the total of 272 species registered in these studies and most of these species are new to Portugal

New Insights on Cistus salviifolius L. Micropropagation

One of the major concerns in the establishment of any mycorrhization program is ensuring the mass production of sterile, consistent and standardized plant material. In the present study, a successful protocol for micropropagation of Cistus salviifolius L. was developed. The process was initiated from nodal segments excised from mature C. salviifolius plant selected due to its mycorrhizal capacities. Murashige and Skoog basal medium supplement with gibberellic acid (0.5 mg/L) and of 6-Benzylaminopurine (0.5 mg/L) was the best medium for proliferation purposes and successful rooting was achieved with the same basal medium supplemented with Indole-3-butyric acid (0.5 mg/L). The proposed methodology represent a novelty because it allowed the rapid multiplication of C. salviifolius starting from mature explants, here reported for the first time, using lower plant growth regulators concentrations than the previously reported for this particular Cistus specie

Terfezia solaris-libera sp. nov., a new mycorrhizal species within The spiny-spored lineages

A new Terfezia species-Terfezia solaris-libera sp. nov., associated with Tuberaria guttata (Cistaceae) is described from Alentejo, Portugal. T. solaris-libera sp. nov. distinct morphology has been corroborated by its unique ITS-rDNA sequence. Macro and micro morphologic descriptions and phylogenetic analyses of ITS data for this species are provided and discussed in relation to similar spiny-spored species in this genus and its putative host plant Tuberaria guttata. T. solaris-libera sp. nov. differs from other spiny-spored Terfezia species by its poorly delimited and thicker peridium and distinct spore ornamentation, and from all Terfezia spp. in it’s ITS nrDNA sequence. In comparison, T. fanfani usually reach large ascocarp dimensions, often with prismatic peridium cells, with olive green tinges in mature gleba and different spore ornamentation. T. lusitanica has a lighter yellowish and thinner peridium and a blackish gleba upon maturity, T. extremadurensis has a thinner well delimited peridium and Tuber-like gleba and T. cistophila has a spermatic odour and is exclusively associated with Cistus spp

Morphological Characterization of the In Vitro Mycorrhizae Formed between Four Terfezia Species (Pezizaceae) with Cistus salviifolius and Cistus ladanifer—Towards Desert Truffles Production in Acid Soils

Terfezia species are obligate symbiotic partners of several xerophytic host plants, mainly belonging to the Cistaceae. Yet, their mycorrhizal associations with members of the genus Cistus remain poorly characterized and their potential application in desert truffle cultivation remains unexplored. This work provides the first anatomic descriptions of the mycorrhizae formed in vitro by four Terfezia species (i.e., T. arenaria; T. extremadurensis; T. fanfani, T. pini) with C. ladanifer and C. salviifolius, two of the most widespread and common Cistus species in acidic soils. All the tested associations resulted in the formation of ectomycorrhizae with well-developed Hartig net, but with a varying degree of mantle development. Our results also demonstrate that all the experimented Terfezia-Cistus combinations expressed high mycorrhization rates. Moreover, the present work shows that C. salviifolius and C. ladanifer are suitable plant hosts for Terfezia species, including some that are, to date, known to be only associated with annual herbs or tree species. This new evidence might aid in broadening the number of situations whereby Terfezia spp. can be cultivated in acid soils.This research was funded by Alentejo 2020 (Project ALT20-03-0145-FEDER-000006) and National Funds through FCT - Foundation for Science and Technology under the Project UIDB/05183/202

Factores ambientais que afectam a riqueza específica de macrofungos em montados de azinho - implicações para a Gestão e Conservação

A riqueza especifica de macrofungos e os mecanismos naturais que a influenciam são ainda pouco conhecidos. Foram utilizados modelos de regressão linear para inferir a relação existente entre a riqueza de macrofungos e diversas variáveis ambientais. De Novembro de 2005 a Abril de 2007 foram amostradas mensalmente as espécies de macrofungos presentes em montados de azinho (Quercus rotundifolia Lam.) no Parque de Natureza de Noudar (Alentejo, Portugal). Verificou-se que a riqueza de espécies micorrízicas aumenta com a classe etária das árvores e com a percentagem de coberto arbustivo, enquanto a riqueza de espécies sapróbias aumenta com as percentagens de cobertos arbustivo e herbáceo, sendo o coberto arbustivo relevante para ambos os grupos tróficos. Pelo exposto anteriormente, propomos que sejam mantidas faixas e pequenas manchas de vegetação arbustiva natural e que o controlo de matos seja efectuado de forma a manter intocada a rizosfera, maximizando desta forma a riqueza de macrofungos. ABSTRACT: Macrofungal richness is still poorly known and the natural mechanisms enhancing macrofungal diversity still remain unclear. We used linear regression models to infer the relationship between mushroom richness and several environmental variables. Therefore, a macrofungal inventory based on fruit bodies was conducted monthly, from November 2005 to April 2007, in holm oak stands (Quercus rotundifolia Lam.) on the Parque de Natureza de Noudar (Alentejo, Portugal). According to our results, mycorrhizal richness increases with tree age class and shrub cover, while saprotrophic richness increases with shrub cover and herbaceous cover. Although, mycorrhizal and saprotrophic richness models differed from each other, results seem to emphasize that shrubs are of the upmost importance for both mycorrhizal and saprotrophic macrofungi in holm oak stands. Thus we propose that strips and patches of natural shrubby vegetation should be maintained and shrub control methods should keep the rhizosphere intact in order to enhance macrofungal richness

Epigeous Macrofungi of the Parque de Natureza de Noudar in Alentejo (Portugal)

This inventory represents the first list of the epigeous macrofungi collected in Parque de Natureza de Noudar (Barrancos, Alentejo, Portugal). Throughout 3 years 162 taxa were registered, from which 8 are new species for Portugal and 77 for Alentejo. Some of them are considered rare species whereas others have been suggested to acquire a conservation status

Serra de São Mamede Natural Park, a macrofungal diversity hotspot in the Mediterranean region

The Serra de São Mamede Natural Park is a biodiversity hotspot in the broad region of Alentejo, Portugal, where the influence of the Atlantic Ocean allows for the continuity of a Eurosiberian-like forest in the heart of a Mediterranean region. The park is therefore unique among the few protected areas in Portugal. We attempt to describe the macrofungal diversity of the park for the first time. Our results reveal a rich and diverse assembly of macrofungal species (239 taxa), and in particular, mycorrhizal species (145 taxa). Furthermore, 26 taxa are newly recorded from Portugal, while 81 taxa are newly recorded from Alto Alentejo Province. Also, the majority of these newly reported taxa were recorded in mixed stands of Quercus pyrenaica Willd.—a European protected habitat of limited extent—whose importance is enhanced now the rich and diverse associated macrofungal community, as shown in our study

Lack of knowledge on ecological determinants and cryptic lifestyles hinder our understanding of Terfezia diversity

Developing below the soil surface desert, truffles are hard to find. Within Terfezia genus, at least 18 species are described and many are endemic to the Mediterranean basin. Ecological and geographic information are key factors for species diagnosis, and so far Terfezia species are believed to be linked to either acidic or basic soils or to specific plant hosts. Thus, we have looked at Terfezia diversity within a relatively homogeneous geographical area in Portugal that is suitable for these species and that covered different soils and different dominant host species. We analyzed the observed intraspecific variability within the context of species ecological preferences (e. g. edaphic and putative host). One of our major findings was the discovery of T.grisea in acid soils in association with Tuberariaguttata, a puzzling information since, until now, this species was only found in alkaline soils. We also report on the linkage of different Terfezia lineages within species and ecologic parameters such as soil texture, soil pH and plant host. Additionally, by placing the collected specimens on the most recent genus phylogeny based on the ITS region, we also updated the number of known Terfezia species occurring in Portugal from three to ten. Terfeziadunensis is here reported for the first time for Portugal. Overall, our results show that the exploration of undersampled sites reveals itself as a good strategy to disclose unknown aspects of desert truffle diversity and ecology. These aspects are of prime importance when considering the economic value of the desert truffles for rural populations in the Mediterranean basin

Lactarius deliciosus isolate UEZB1 internal transcribed spacer 1, partial sequence; 5.8S ribosomal RNA gene and internal transcribed spacer 2, complete sequence; and 28S ribosomal RNA gene, partial sequence

LOCUS JQ066791 439 bp DNA linear PLN 20-MAR-2012 DEFINITION Lactarius deliciosus isolate UEZB1 internal transcribed spacer 1, partial sequence; 5.8S ribosomal RNA gene and internal transcribed spacer 2, complete sequence; and 28S ribosomal RNA gene, partial sequence. ACCESSION JQ066791 VERSION JQ066791.1 GI:380467948 KEYWORDS . SOURCE Lactarius deliciosus ORGANISM Lactarius deliciosus Eukaryota; Fungi; Dikarya; Basidiomycota; Agaricomycotina; Agaricomycetes; Russulales; Russulaceae; Lactarius