Between 2006 and 2007, palm trees growing in both gardens and public parks and natural palm groves in the Canary Islands ( Spain ), and showing symptoms of wilt and dieback, were surveyed. Isolates were recovered from affected tissues of the crowns, leaves and vascular fragments on potato dextrose agar (PDA). After incubation, the Fusarium spp. colonies recovered were single-spored. They were transferred to PDA and Spezieller Nahrstoffarmer Agar (SNA) for morphological identification. Identification of Fusarium oxysporum f. sp. Canariensis was confirmed by PCR with the specific primers HK66 and HK67, which amplified a fragment of 567 bp. Fusarium wilt caused by F. oxysporum f. sp. canariensis was found on 54 Phoenix canariensis trees growing on four islands: Gran Canaria, Fuerteventura, La Palma and Tenerife. F. proliferatum occurred on fifteen palms (10 P. canariensis , 1 P. dactylifera , 3 Roystonea regia and 1 Veitchia joannis ) located in Gran Canaria, Fuerteventura and Tenerife . Both these Fusarium species were found only in diseased palms from gardens and public parks, but not in natural palm groves. The results show that Fusarium wilt of P. canariensis is common in the Canary Islands and for the first time report F. proliferatum affecting different palm species in those islands.
Este trabajo se ha realizado con financiacion del Proyecto AGL2007-64690 (Ministerio de Educacion y Ciencia, Fondos FEDER).
Agradecemos el apoyo de E. Landeras de Consejeria de Agroganaderia y Recursos Autoctonos del Principado de Asturias, a L. Alvarez, A. Roman, O. Martinez, V. Garrigues de la Universidad Politecnica de Valencia y a todos los viveristas que han participado en estos estudios.
The susceptibility of the grapevine rootstocks most commonly used in Spain to black-foot (Ilyonectria liriodendri and Dactylonectria macrodidyma-complex) and petri disease (Cadophora luteo-olivacea, Phaeomoniella chlamydospora and five species of Phaeoacremonium) pathogens was evaluated. Rooted cuttings of rootstocks 110R, 1103P, 140Ru, 161-49C, 196-17C, Fercal and SO4 were inoculated with black-foot pathogens by dipping their roots in conidial suspensions (5×105 conidia mL-1), planted in pots containing sterilized peat moss and placed in a greenhouse. After four months of incubation, root disease severity index and dry weights of shoots and roots were recorded for each plant. Regarding petri disease pathogens, one-year-old grapevine cuttings of five rootstocks (41B, 140Ru, 161-49C, 1103P and 110R) were vacuum-inoculated with spore suspensions (106 conidia mL-1) of the fungal species and planted in two fields. After four months, the proportion of vines that sprouted in spring was visually determined. At the end of the growing season, dormant plants were uprooted, washed, and assessed for undried shoot weight. Then, the stem of each grapevine cutting was transversally split at 10 cm from the base of the plant to estimate the percentage of vascular tissue discoloured on a scale of 0 to 4. All rootstocks inoculated with Ilyonectria and Dactylonectria species were affected by the disease in some degree, the rootstock 110R being the most susceptible to black-foot. Petri disease pathogens caused a significant reduction of sprouting and shoot weight, as well as a significant increase of disease severity percentage in all grapevine rootstocks with the exception of 161-49C. Both 110R and 140Ru were the most susceptible. Grapevine rootstocks showed different levels of disease resistance, being the rootstock 110R the most susceptible to both diseases and the 161-49C rootstock the most tolerant to petri disease infection.
Phytophthora ramorum has been detected in official plant health surveys on Rhododendron , Viburnum and Camellia in ornamental nurseries in northern Spain since 2003. A collection of 94 isolates of P. ramorum was obtained from 2003 to 2008 from plants with symptoms at different geographical locations. Isolates were identified based on morphology and sequence of the rDNA ITS region. Mating type, genetic variation, sensitivity to phenylamide fungicides and aggressiveness of these isolates were determined. All isolates belonged to the A1 mating type, ruling out the possibility of genetic recombination. Seven microsatellite markers were used to study genetic diversity; three out of the seven microsatellite markers were polymorphic within the Spanish population of P. ramorum . This study confirms that all Spanish isolates of P. ramorum belonged to the EU1 lineage. Twelve intralineage genotypes were detected, five that are unique to Spain (EU1MG38, EU1MG41, EU1MG37, EU1MG39 and EU1MG40) and seven that are also present in at least one other European country (EU1MG1, EU1MG29, EU1MG22, EU1MG13, EU1MG2, EU1MG18 and EU1MG26). Genotypes EU1MG37, EU1MG39 and EU1MG40 were isolated from Rhododendron from one region; EU1MG38 and EU1MG41 were isolated from Camellia from two different regions. Isolates of genotype EU1MG38 were resistant to metalaxyl and mefenoxam. The level of genetic diversity within the Spanish population of P. ramorum is limited and indicates a relatively recent clonal expansion.
Pythium indigoferae and Pythium irregulare, identified based on morphological and physiological characteristics, were isolated from necrotic roots, crown tissues and the rhizosphere of apple trees in Tunisia from 23 apple orchards in spring and autumn 2007–2009. The virulence assays on excised twigs, using different Pythium species isolated demonstrated that these oomycetes were pathogenic on the Anna, Lorka and Meski varieties and the MM106 rootstock. However, the biggest lesion area was noted on MM106 rootstock. Thus, it appeared that this rootstock is more susceptible to Pythium infections than Anna, Meski and Lorka apple varieties. Furthermore, it is important to note that in vitro tests showed that P. indigoferae seems to be more virulent than P. irregulare.
The diversity of Phytophthora species in declining Fagaceae forests in Europe is increasing in the last years. The genus Quercus is one of the most extended Fagaceae genera in Europe, and Q. ilex is the dominant tree in Spain. The introduction of soil-borne pathogens, such as Phytophthora in Fagaceae forests modifies the microbial community present in the rhizosphere, and has relevant environmental and economic consequences. A better understanding of the diversity of Phytophthora spp. associated with Q. ilex is proposed in this study by using Next Generation Sequencing (NGS) in six Q. ilex stands located in three regions in Spain. Thirty-seven Phytophthora phylotypes belonging to clades 1 to 12, except for clades 4, 5 and 11, are detected in this study, which represents a high diversity of Phytophthora species in holm oak Spanish forests. Phytophthora chlamydospora, P. citrophthora, P. gonapodyides, P. lacustris, P. meadii, P. plurivora, P. pseudocryptogea, P. psychrophila and P. quercina were present in the three regions. Seven phylotypes could not be associated with known Phytophthora species, so they were putatively named as Phytophthora sp. Most of the detected phylotypes corresponded to terrestrial Phytophthora species but aquatic species from clades 6 and 9 were also present in all regions.
Fungal pathogens are able to produce inoculum (spores) on plant debris present on the soil surface of vineyards.
These spores can then reach plant surfaces and cause severe grapevine infections when environmental conditions are favourable. The capacity of plant diversity to increase the resistance of crops towards pests and invasive species is very well-known. For instance, Brassica spp. have been already investigated for their capacity to effectively suppress soil-borne inoculum of some causal agents of Black-foot disease in grapevines in vineyard soils. It may also have positive effect on the some dagger nematodes. Cover crops also stimulate the development of microbial communities such as arbuscular mycorrhizal fungi. Many management strategies have been developed against these important grapevine pathogens, but the effects of soil cover vegetation or organic mulching against spore dispersal, acting as a barrier, have been scarcely explored.
Thus, in the BIOVINE project (www.biovine.eu) specific experiments were planned in order to verify the possibility
of using cover crops: i) to control some relevant pathogens producing inoculum (spores) on plant debris present on the soil surface of vineyards; ii) to determine the presence of causal agents of Petri disease of grapevines on the roots of cover crops; iii) to promote mykorrhizal communities associated with grapevine roots; iv) to control arthropod pests (repellent of arthropods or attracting beneficials); v) to investigate Brassica plants effect on the soil-borne pest nematode Xiphinema index.