Stone pine ( Pinus pinea L.) is an economically important forest tree in Mediterranean climates and has been the target for selection efforts through micropropagation. Previous attempts on microshoots, derived from mature seed cotyledons, reached incipient rooting after induction with a combination of auxin and hypertonic shock, but their development in vitro was not sustained. At this stage, co‐culturing plantlets with some fungi isolated from ectomycorrhizas succeeded in overcoming this barrier, enabling satisfactory development in vermiculite and later in soil. About half of the fungal isolates tested helped the plants resume root growth. Although control plants (in the absence of fungi) developed roots at a later stage, i.e. during the post‐transplanting acclimation in vermiculite, their growth was weaker. The root systems of some inoculated plants had ectomycorrhizas from the introduced fungi being carried over when the plants were transferred from the co‐cultures to vermiculite. In conclusion, co‐culturing rooted microshoots with ectomycorrhizal fungi can be an effective means to overcome the difficulties encountered in the use of micropropagation methods on this species.

ISHS III International Symposium on Acclimatization and Establishment of Micropropagated Plants EFFECTS OF CARBON SOURCE, CARBON CONCENTRATION AND CULTURE CONDITIONS ON IN VITRO ROOTING OF PINUS PINEA L. MICROSHOOTS

Le clade Amidella en Europe (Basidiomycota: Amanitaceae): clarification du controversé Amanita valens (E.-J.Gilbert) Bertault et importance de l'utilisation de primers PCR spécifiques aux taxons pour l'identification. Les espèces de Amanita ser. Amidella (E.-J.Gilbert) Neville & Poumarat forment un clade bien défini, mais certains taxons restent difficiles à discriminer. En particulier, le concept d'Amanita valens (E.-J.Gilbert) Bertault reste controversé. Pour comprendre le placement phylogénétique d'un ensemble de collections du sud du Portugal avec un nouveau barcode nrDNA, nous avons obtenu des séquences nrDNA pour des collections type non évaluées auparavant. Le taxon précédemment décrit comme A. curtipes f. pseudovalens Neville & Poumarat est interprété comme une espèce distincte, A. pseudovalens comb. nov., stat. nov., et est génétiquement indistinct de ces collections portugaises, clarifiant ainsi leur contexte taxonomique. Cependant, la morphologie et l'écologie de nos collections sont distinctes (respectivement, basidiospores ellipsoïdes à oblongues et occurrence sur des sols schisteux), et sont proposées comme une nouvelle variété, Amanita pseudovalens var. tartessiana var. nov. Ces développements permettent également de mieux diagnostiquer les taxons d'Amidella en Europe, progrès décisif pour les récoltes de la fin de l'hiver au printemps, et une clé d'identification est proposée. Malgré tout, la cooccurrence et la similitude morphologique de la nouvelle variété avec la très appréciée et comestible A. ponderosa Malençon & R.Heim pourrait laisser certaines collections non résolues. Ainsi, une approche de marqueurs moléculaires a été développée, afin de fournir une aide claire et économique à l'identification pour complémenter la clé. Les outils de diagnostic proposés peuvent être appliqués à une révision de la chorologie des taxons d'Amidella européens à partir des enregistrements existants, des matériaux conservés et des collections futures.

Species determination in the Amidella clade is notoriously difficult, because of the relative dearth of diagnostic characters and the rather common occurrence of homoplasies. This results in a substantial number of misnamed and unnamed collections, a misapprehension of the geographic range of known species and a gross underestimation of the number of species it contains. To assess the diversity that should be considered as part of Amidella , DNA sequences available for this group were retrieved in public nucleotide databases, using a combination of approaches to achieve a comprehensive representation. Phylogenetic analysis based on the aligned ITS sequences, consistently with the results from other molecular markers (ncLSU, RPB2 , TEF1 , BTUB ), suggests five major clades: one containing the type species Amanita volvata ; another for Amanita ponderosa and allies; a third one (roughly half of all species) with Amanita lepiotoides ; and two others without valid species yet. At species level, around 81 clades were delimited, of which only 16–17 can be assigned a valid name, with a few more corresponding to provisional taxa listed in the amanitaceae.org website. Up to three further species without assigned sequences might correspond to the proposed clades. The current evidence suggests a rather narrow geographic range for most of these clades. This study provides a phylogeny-arranged outlook of the worldwide distribution of Amidella species and an infrasectional framework for optimising taxonomic sampling and designing clade-specific molecular markers to assist in identification. Graphical Abstract