Evaluation of Insect Associated and Plant Growth Promoting Fungi in the Control of Cabbage Root Flies

Delia radicum L. or cabbage maggot is an important pest for Brassicaceous crops. There are currently no registered chemical control agents for its control in Slovenia. Fungal control agents for cabbage maggot were therefore sought among nine rhizosphere-compatible and plant growth-promoting, soil-adapted, and entomopathogenic species to cabbage maggots and were assayed in in vitro and soil laboratory bioassays. In the in vitro tests, the conidial suspensions were applied directly to cabbage maggot eggs. The soil tests mimicked pathways of natural exposure of various insect life stages to the fungal strains. Conidial concentrations used in soil tests were comparable to economic rates for in-furrow application. The following fungi were tested: Trichoderma atroviride P. Karst. (2 isolates), Trichoderma koningiopsis Samuels, C. Suárez & H.C. Evans (1), Trichoderma gamsii Samuels & Druzhin. (3), Beauveria brongniartii (Saccardo) Petch (1), Beauveria bassiana (Balsamo-Crivelli) Vuillemin (2), Metarhizium robertsii J.F. Bisch., Rehner & Humber (1), Metarhizium anisopliae (Metschn.) Sorokin (4), Purpureocillium lilacinum (Thom) Luangsa-ard, Houbraken, Hywel-Jones & Samson (2), and Clonostachys solani f. nigrovirens (J.F.H. Beyma) Schroers (2). Abbott's corrected mortality in the in vitro tests ranged from 0.0 ± 18.9 to 47.6 ± 9.0% and in the soil test from 2.4 ± 13.0 to 68.2 ± 21.5%. Seven isolates (B. bassiana [isolate 1174], C. solani [1828], M. anisopliae [1154 and 1868], T. atroviride [1872], T. koningiopsis [1874], and T. gamsii [1876]) caused significant cabbage maggot mortality in either in vitro or soil tests. The importance of fungal ecology as a criterion during the screening of potential biological control agents is discusse

ReSolVe project – Restoring optimal Soil functionality in degraded areas within organic Vineyards

In both conventional and organic European vineyards, it is quite common to have areas characterized by problems in vine health, grape production and quality. These problems are very often related to sub-optimal soil functionality, caused by an improper land preparation before vine plantation and/or management. Different causes for soil malfunctioning can include: poor organic matter content and plant nutrient availability (both major and trace elements); imbalance of some element ratios (Ca/Mg, K/Mg, P/Fe, and Fe/Mn); pH; water deficiency; soil compaction and/or scarce oxygenation. Fertility related problems can often be compensated in conventional settings with externally introduced fertilizers that are not permitted in organic vineyards. ReSolVe is a transnational and multidisciplinary research project aimed at testing the effects of selected agronomic strategies for restoring optimal soil functionality in degraded areas within organic vineyard. The term "degraded areas within vineyard" means areas showing reduced vine growth, disease resistance, grape yield and quality. These areas may have lost their soil functionality because of either an improper land preparation, or an excessive loss of soil organic matter and nutrients, erosion and/or compaction. The project, financed by Core-Organic plus program of the ERA-NET plus action (2015-2018), aims at identifying the main causes of the soil functionality loss and testing different organic recovering methods. The different restoring strategies will implement: i) compost, ii) green manure with winter legumes, iii) dry mulching with cover crops. The strategies will be tested according to their efficiency to improve: i) plant and roots growth and well-being; ii) grape yield and quality; iii) quality of soil ecosystem services and their stability over the years; iv) better express of the “terroir effect”, that is, the linkage of wine quality to the environmental characteristics of the cultivation site. The project involves 8 research groups in 6 different EU countries (Italy, France, Spain, Sweden, Slovenia, and Turkey), with experts from several disciplines, including soil science, ecology, microbiology, grapevine physiology, viticulture, and biometry. The experimental vineyards are situated in Italy (Chianti hills and Maremma plain, Tuscany), France (Bordeaux and Languedoc), Spain (La Rioja) and Slovenia (Primorska) for winegrape, and in Turkey (Adana and Mersin) for table grape. The restoration techniques and the monitoring methodologies developed and tested during the ReSolVe project will be described in specific final guidelines. The restoration techniques will be accessible for all the European farmers and will be low cost and environmental-friendly. A protocol of analyses and measurements between the all partners will allow an effective and comparable monitoring of vineyard ecosystemic functioning in European countries

Response of African eggplants to Fusarium spp. and identification of sources of resistance

Eggplant (Solanum spp.) production in Arumeru district and other parts of Africa is severely affected by wilting diseases of unknown etiology. Fusarium spp. characterized through morphological and sequence analysis of the translation elongation factor associated with Fusarium wilt of eggplants was used to test the response of three different eggplant species. Three Solanum spp. accessions were tested in a screen house at the seedling stage for resistance to two isolates each of Fusarium equiseti (corda) Sacc, Fusarium solani (Mart.) Sacc and Fusarium oxysporum (Schlecht). The study indicated that accessions MM 1131 (Solanum macrocapon) and N 19 (Solanum anguivi) accessions are susceptible to F. equiseti. Accession N 19 (S. anguivi) was susceptible to F. solani while both N 19 (S. anguivi) and MM 1131 (S. macrocarpon) was also susceptible to F. oxysporum f. sp. melongenae. Ninety-three accessions of cultivated and wild eggplants were subsequently evaluated in two screen house trials for resistance to Fusarium wilt. A root dip technique was used to inoculate the accessions with isolate Fs 40 (F. oxysporum f.sp. melongenae). Seventeen of the 93 accessions were found to be resistant and they belonged to Solanum macrocarpon and Solanum aethiopicum species. Accessions in S melongena were found to be the most susceptible. Eggplant accessions that showed high levels of resistance could potentially serve as valuable sources of Fusarium wilt resistance in eggplant breeding programs in Tanzania and beyond.Keywords: African eggplants, Fusarium spp. susceptibility, resistanc

Out in the Cold: Identification of Genomic Regions Associated With Cold Tolerance in the Biocontrol Fungus Clonostachys rosea Through Genome-Wide Association Mapping

There is an increasing importance for using biocontrol agents in combating plant diseases sustainably and in the long term. As large scale genomic sequencing becomes economically viable, the impact of single nucleotide polymorphisms (SNPs) on biocontrol-associated phenotypes can be easily studied across entire genomes of fungal populations. Here, we improved a previously reported genome assembly of the biocontrol fungus Clonostachys rosea strain IK726 using the PacBio sequencing platform, which resulted in a total genome size of 70.7 Mbp and 21,246 predicted genes. We further performed whole-genome re-sequencing of 52 additional C. rosea strains isolated globally using Illumina sequencing technology, in order to perform genome-wide association studies in conditions relevant for biocontrol activity. One such condition is the ability to grow at lower temperatures commonly encountered in cryic or frigid soils in temperate regions, as these will be prevalent for protecting growing crops in temperate climates. Growth rates at 10°C on potato dextrose agar of the 53 sequenced strains of C. rosea were measured and ranged between 0.066 and 0.413 mm/day. Performing a genome wide association study, a total of 1,478 SNP markers were significantly associated with the trait and located in 227 scaffolds, within or close to (< 1000 bp distance) 265 different genes. The predicted gene products included several chaperone proteins, membrane transporters, lipases, and proteins involved in chitin metabolism with possible roles in cold tolerance. The data reported in this study provides a foundation for future investigations into the genetic basis for cold tolerance in fungi, with important implications for biocontrol

Neonectria liriodendri sp. nov., the main causal agent of black foot disease of grapevines

Black foot disease is a serious disease of grapevine crops in most areas where vines are grown. Mainly two species of Cylindrocarpon, C. destructans and C. macrodidymum, are associated with this disease. Recent studies have revealed a tremendous molecular variation within the former but only slight molecular variation within the latter, indicating that C. destructans presents a complex of several species The present study elucidates the taxonomic status of C. destructans-like isolates associated with black foot disease of grapevines. Grapevine isolates were studied morphologically, subjected to DNA analyses of their ITS and partial β-tubulin genes, and were mated in all combinations in vitro. Cylindrocarpon destructans strains isolated from grapevines in Europe and South Africa appeared morphologically and genetically identical, and had identical ITS and partial β-tubulin gene sequences. Phylogenetic analyses placed these strains in a clade closely related but clearly distinct from other clades with C. destructans-like anamorphs obtained from various herbaceous or woody hosts. Only the ex-type strain of Cylindrocarpon liriodendri had identical sequences to strains isolated from grapevines, and could also not be distinguished by morphological characters. The grapevine isolates are therefore reidentified here as Cylindrocarpon liriodendri. Cylindrocarpn liriodendri formed perithecia in heterothallic conditions and the holomorph of this species is described as Neonectria liriodendri sp. nov. Neonectria liriodendri is genetically distinct from the ex-type strain of Neonectria radicicola, which originated from Cyclamen in Sweden. Both ex-type strains also differ from at least two other clades comprising additional C. destructans-like strains. Many of these strains originated from Panax sp., which is the host of the type of C. destructans. Our phylogenetic analyses indicate that C. destructans is not the anamorph of N. radicicola and that N. liriodendri, N. radicicola and several C. destructans-like taxa may have evolved independently within the same phylogenetic species complex

The Trichoderma koningii aggregate species

The morphological concept of Trichoderma koningii is found to include several species that differ from each other in details of phenotype (including conidium morphology, growth rate) and biogeography. Phylogenetic analysis utilizing partial sequences of the translation-elongation factor 1 alpha (tef1), as well as fragments of actin and calmodulin genes, indicate that phenotypic characters typical of T. koningii evolved independently in three well-separated main lineages. Combined molecular and phenotype data lead to the development of a taxonomy with the recognition of twelve taxonomic species and one variety within the three lineages. These lineages include: (1) T. koningii and T. ovalisporum and the new species T. caribbaeum var. caribbaeum, T. caribbaeum var. aequatoriale, T. dorotheae, T. dingleyae, T. intricatum, T. koningiopsis, T. petersenii and T. taiwanense; (2) the new species T. rogersonii and T. austrokoningii, and (3) the new anamorph T. stilbohypoxyli

Production of Obionin A and Derivatives by the Sooty Blotch Fungus Microcyclospora malicola.

A multitude of sooty blotch and flyspeck fungi, mainly belonging to the Ascomycetes order Capnodiales, causes dark blemishes and flyspeck-like spots on apples worldwide. Different sooty blotch and flyspeck fungi can coexist in the same orchard and even on a single fruit. Our preceding experiments revealed an activity of Microcyclospora malicola strain 1930 against the anthracnose fungus Colletotrichum fioriniae in dual culture assays. Extracts of M. malicola strain 1930 showed a broad bioactivity against filamentous fungus Mucor hiemalis and gram-positive bacterium Bacillus subtilis. A bioactivity-guided isolation led to the identification of obionin A (1) as the main active principle. In addition to 1, which was previously isolated from the marine fungus Leptosphaeria obiones, we isolated three derivatives. Metabolite 2 bears a keto function at C-6, besides the replacement of oxygen by nitrogen at position 10. Two more derivatives are adducts (3, 4) of acetone as work-up artifacts. Because obionin A (1) and its derivative 2 showed cytotoxic effects and antifungal activities, we propose a role of these secondary metabolites in the antagonism between M. malicola and other apple colonizing sooty blotch and flyspeck fungi, other epiphytes, or apple pathogens competing for the same ecological niche