Bacteria associated with the rhizosphere of manganese-deficient date palms affected by Brettle Leaf Disease

[EN] Brittle leaf disease or "Maladie des feuilles cassantes" (MFC) is a disorder affecting date palm (Phoenix dactylifera) orchards in oases of southern Tunisia. No causal pathogen has yet been identified, despite search for nematodes, fungi, bacteria, viruses and viroids. Although the soils of the oases in which affected palms grow have similar contents of total and exchangeable manganese (Mn) compared with oases with no disease, the leaflets of affected palms are deficient in this element. Here, we characterized the bacterial flora in the rhizosphere of three MFC-affected date palms and three non-affected controls. Our results show a high bacterial diversity in the date palm rhizosphere. From 201 clones corresponding to a 16S rDNA PCR fragment, we identified 177 different bacterial phylotypes. Some of them corresponded potentially to Mn-oxidizing bacteria that may be involved in soil Mn oxidation and, consequently, deplete this micronutrient in MFC-affected date palms. More detailed studies are necessary to establish the exact role of these bacteria in the disease syndrome.Work supported by grants AGL2007-65653-C02-01 (JAD) and AGL2007-65653-C02-02 (NDV) from Ministerio de Educacion y Ciencia (MEC) from Spain and ACOMP07/261 (JAD) from Generalitat Valenciana. JM was the recipient of a predoctoral fellowship from MEC. We thank Holly Kuzmiak-Ngiam (University of Rochester, USA) for critical reading of the manuscript.Marques, J.; Duran Vila, N.; Namsi, A.; Bove, J.; Daros Arnau, JA. (2012). Bacteria associated with the rhizosphere of manganese-deficient date palms affected by Brettle Leaf Disease. Journal of Plant Pathology. 94(1):157-169. https://doi.org/10.4454/jpp.fa.2011.004S15716994

Manganese Deficiency is Associated with Histological Changes in Date Palm Fronds Showing Brittle Leaf Disease Symptoms

Several diseases, pests and physiological disorders threat date palm life. Among them, Brittle leaf disease or, in French, Maladie des feuilles cassantes (MFC) is a lethal disease that has acquired alarming proportions since the 1960s, when it was first observed in southern Tunisia. Up to now no causal agent has been found. Previous studies have reported that brittle fronds are deficient in manganese but the mechanisms involved in symptom expression have not yet been identified. In this study, an anatomical investigation was carried out in order to identify the deleterious effect of MFC on the development and structure of palm fronds. Results showed that in old healthy fronds the major tissue is schlerenchyma, which is characterized by very thick lignified cell walls and is often associated with conducting tissue. Indeed, large vessels are surrounded by schlerenchyma fibers that are strongly lignified when compared with those of young fronds. Diseased young fronds have almost the same shape as the healthy young fronds. Nevertheless, in old diseased fronds MFC induces important structural changes. Xylem and phloem vessels are larger, taking more space through the fibers that are smaller in number and size, and tighter. The larger diameter of the lumen and the lower thickness of the fibers present in the fronds in advanced disease stages, decreases the lignin content of cells walls, leading to an increased frond friability, which gives "the brittle leaf disease" its name

Phylogeny and mycotoxin profile of pathogenic fusarium species isolated from sudden decline syndrome and leaf wilt symptoms on date palms (Phoenix dactylifera) in tunisia

In 2017–2018, extensive symptoms of sudden decline and fruit rot were observed on date palms in southern Tunisia. Samples of diseased plants were randomly collected in six localities. Based on morphological identification, Fusarium was the most frequent fungal genus detected. A sequencing of translation elongation factor, calmodulin, and second largest subunit of RNA polymerase II genes was used to identify 63 representative Fusarium strains at species level and investigate their phylogenetic relationships. The main species detected was Fusarium proliferatum, and at a much lesser extent, Fusarium brachygibbosum, Fusarium caatingaense, Fusarium clavum, Fusarium incarnatum, and Fusarium solani. Pathogenicity on the Deglet Nour variety plantlets and the capability to produce mycotoxins were also assessed. All Fusarium species were pathogenic complying Koch’s postulates. Fusarium proliferatum strains produced mainly fumonisins (FBs), beauvericin (BEA), and, to a lesser extent, enniatins (ENNs) and moniliformin (MON). All F. brachygibbosum strains produced low levels of BEA, diacetoxyscirpenol, and neosolaniol; two strains produced also T-2 toxin, and a single strain produced HT-2 toxin. Fusarium caatingaense, F. clavum, F. incarnatum produced only BEA. Fusarium solani strains produced MON, BEA, and ENNs. This work reports for the first time a comprehensive multidisciplinary study of Fusarium species on date palms, concerning both phytopathological and food safety issues

Phylogeny and mycotoxin profiles of pathogenic Alternaria and Curvularia species isolated from date palm in southern Tunisia

Date palm (Phoenix dactylifera L.), is a widely cultivated crop across North Africa, with about 300 thousand tons of fruits produced per year, in Tunisia. A wide range of fungal pathogens has been associated with leaf spots of date palm, Alternaria species being the most frequently reported. Symptomatic leaves of Deglet Nour variety were randomly collected in six localities in Tunisia. We used a polyphasic approach to identify 45 Alternaria and five Curvularia strains isolated from date palm, confirming their pathogenicity. Sequencing of allergen Alt-a1, glyceraldehyde-3-phosphate dehydrogenase (gpd) and calmodulin genes allowed us to group 35 strains in Alternaria Section, and 10 strains in Ulocladioides section. Based on sequencing analyses of Internal Transcribed Spacer, gpd and elongation factor genomic regions, all Curvularia strains were identified as Curvularia spicifera. All Alternaria and Curvularia species tested on date palm plantlets proved to be pathogenic, fulfilling Koch's postulates. Although no significant differences were observed among the species, the highest mean disease severity index was observed in A. arborescens, while the lowest corresponded to C. spicifera. The capability of these strains to produce mycotoxins in vitro was evaluated. None of the A. consortialis strains produced any known Alternaria mycotoxin, whereas more than 80% of the strains included in Alternaria section Alternaria produced variable amounts of multiple mycotoxins such as alternariol, alternariol monomethyl ether, altenuene, tenuazonic acid and tentoxin. Curvularia spicifera strains produced detectable traces of fumonisins B. This work reports a first comprehensive multidisciplinary study of mycotoxigenic Alternaria species and C. spicifera associated with leaf spot disease on date palm

Diagnosis of "maladie des feuilles cassantes" or brittle leaf disease of date palms by detection of associated chloroplast encoded double stranded RNAs

The “Maladie des feuilles cassantes” (MFC) or “Brittle leaf disease” of date palms is associated with the accumulation of two populations of small, chloroplast-encoded RNAs. A plasmid vector containing a cDNA with partial sequences of both of these RNA populations was used to synthesize a DIG-labeled bifunctional probe by PCR. The probe has been tested to detect, by molecular hybridization, MFC-associated RNAs from dsRNA-enriched palm leaflet preparations. Leaflet samples from MFC-affected date palm trees consistently gave a positive hybridization signal regardless of the date palm cultivar, severity of symptoms, or geographical location, whereas samples from date palm trees affected by other biotic and abiotic stresses tested negative. The assay is specific for MFC and can be used for early diagnostic purposes