Giovanni Cafà

The banana skipper, Erionota torus Evans (Lepidoptera, Hesperiidae, Hesperiinae, Erionotini) is a South-east Asian pest of banana that, in the last 60 years, has spread to the southern Philippines, Taiwan, Japan, India, Sri Lanka, Mauritius and La Réunion, and potentially threatens Africa and Tropical America. A partial library of DNA barcodes from the indigenous and introduced ranges was built. Based on our analysis, the indigenous populations can be divided into an ‘East’ group, in China and Vietnam, and a ‘West’ group in India, Nepal, Myanmar and west Malaysia. Further, within the ‘West’ group, there is a coherent ‘Malaysia’ subgroup from west Malaysia. Introduced populations in south India, La Réunion and Taiwan showed almost no variation in barcodes, suggesting they are each based on a single homogenous introduction. We conclude that the introduced populations in Taiwan and Japan match the ‘East’ group, the introduced populations in Mauritius and La Réunion match the ‘Malaysia’ subgroup and the introduced population in south India matches the ‘West’ group. These results are discussed in the context of existing ideas regarding the source of each introduction, and the implications in terms of pathways of entry.

Phalaena militta Stoll, [1781], currently in the combination Thyrgis militta, is transferred to the new combination Calodesma militta. Phalaena militta is the type species of Thyrgis Walker, 1854, and so Thyrgis is a junior synonym of Calodesma Hübner, [1820]. The reinstated genus Seileria Dognin, 1923 is the next available name for the genus previously known as Thyrgis, and the remaining eight species and their subspecies currently in Thyrgis are transferred to new combinations as species of Seileria: S. angustifascia (Hering, 1925), S. basipunctata (Hering, 1926), S. constrictifascia (Dognin, 1919), S. flavonigra (Dognin, 1910), S. investigatorum (Toulgoët, 1988), S. marginata (Butler, 1875), S. meres (Druce, 1911), S. phlegon (Druce, 1885), S. phlegon ruscia (Druce, 1895), S. tenuifascia (Hering, 1930) and S. tenuifascia daguana (Hering, 1930). Eucyanoides Toulgoët, 1988, currently a synonym of Thyrgis, is made a new subjective synonym of Seileria. Based on DNA barcodes, we recognise three very similar, sexually dimorphic and in two cases polymorphic South American species of Calodesma with some phenotypes in common but very similar male genitalia: C. militta (BOLD:AAK1660), C. sp. cf. collaris (BOLD:ABZ2392) and C. pseudocollaris Cock new species (BOLD:AEI2170). Calodesma militta is widespread in South America, with two male morphs (collaris and dioptis) and two female morphs with variable markings (white and orange morphs). Centronia plorator Kaye, [1923] and Thyrgis lacryma Dognin, 1919 are variants of the white female morph and are new synonyms of Calodesma militta. A third female morph with red markings was not sequenced and could not be allocated to a species. Calodesma sp. cf. collaris (BOLD:ABZ2392) occurs in southern South America with both male morphs but only a white female morph. Calodesma pseudocollaris new species (BOLD:AEI2170) is only known from Trinidad, with one male morph (collaris) and the white female morph. Although more than ten morphs relating to this complex have been described as species, they cannot be synonymised without more data on distribution of the different species or DNA barcodes from the type specimens. Collated life history information indicates species of this group are split between Malpighiaceae feeders and Bromeliaceae feeders, but more work is needed to define these differences. The morphism patterns observed are discussed in terms of Müllerian mimicry and mimicry rings, and we suggest that in Trinidad (and elsewhere) there is a loose mimicry ring of diurnal black species with white spots or transparent patches on the wings which are most conspicuous and frequently observed when feeding on white Asteraceae flowers.

Following reports of an invasive snail causing crop damage in the expansive Mwea irrigation scheme in Kenya, samples of snails and associated egg masses were collected and sent to CABI laboratories in the UK for molecular identification. DNA barcoding analyses using the cytochrome oxidase subunit I gene gave preliminary identification of the snails as Pomacea canaliculata, widely considered to have the potential to be one of the most invasive invertebrates of waterways and irrigation systems worldwide and which is already causing issues throughout much of south-east Asia. To the best of our knowledge, this is the first documented record of P. canaliculata in Kenya, and the first confirmed record of an established population in continental Africa. This timely identification shows the benefit of molecular identification and the need for robust species identifications: even a curated sequence database such as that provided by the Barcoding of Life Data system may require additional checks on the veracity of the underlying identifications. We found that the egg mass tested gave an identical barcode sequence to the adult snails, allowing identifications to be made more rapidly. Part of the nuclear elongation factor 1 alpha gene was sequenced to confirm that the snail was P. canaliculata and not a P. canaliculata/P. maculata interspecies hybrid. Given the impact of this species in Asia, there is need for an assessment of the risk to Africa, and the implementation of an appropriate response in Kenya and elsewhere to manage this new threat to agriculture and the environment.

The genus Stagonosporopsis is classified within the Didymellaceae family and has around 40 associated species. Among them, several species are important plant pathogens responsible for significant losses in economically important crops worldwide. Stagonosporopsis vannaccii is a newly described species pathogenic to soybean. Here, we present the draft whole-genome sequence, gene prediction, and annotation of S. vannaccii isolate LFN0148 (also known as IMI 507030). To our knowledge, this is the first genome sequenced of this species and represents a new useful source for future research on fungal comparative genomics studies.

Background: Himalayan balsam Impatiens glandulifera Royle (Balsaminaceae) is a highly invasive annual species native of the Himalayas. Biocontrol of the plant using the rust fungus Puccinia komarovii var. glanduliferae is currently being implemented, but issues have arisen with matching UK weed genotypes with compatible strains of the pathogen. To support successful biocontrol, a better understanding of the host weed population, including potential sources of introductions, of Himalayan balsam is required. Methods: In this molecular study, two new complete chloroplast (cp) genomes of I. glandulifera were obtained with low coverage whole genome sequencing (genome skimming). A 125-year-old herbarium specimen (HB92) collected from the native range was sequenced and assembled and compared with a 2-year-old specimen from UK field plants (HB10). Results: The complete cp genomes were double-stranded molecules of 152,260 bp (HB92) and 152,203 bp (HB10) in length and showed 97 variable sites: 27 intragenic and 70 intergenic. The two genomes were aligned and mapped with two closely related genomes used as references. Genome skimming generates complete organellar genomes with limited technical and financial efforts and produces large datasets compared to multi-locus sequence typing. This study demonstrates the suitability of genome skimming for generating complete cp genomes of historic herbarium material. It also shows that complete cp genomes are solid genetic markers for population studies that could be linked to plant evolution and aid with targeting native range and natural enemy surveys for biocontrol of invasive species.

The fall armyworm, Spodoptera frugiperda, a moth originating from the American continent, has recently invaded most African countries, where it is seriously threatening food security as a pest of cereals. The current management methods rely heavily on the use of synthetic insecticides but there is a need for more sustainable control methods, including biological control. Surveys were conducted in two West African countries, Ghana and Benin, to determine the native parasitoid complex and assess parasitism rates of S. frugiperda. Samples of S. frugiperda eggs and larvae were collected in maize fields located in 56 and 90 localities of Ghana and Benin, respectively, from July 2018 to July 2019. Ten species were found parasitizing the pest, including two egg parasitoids, one egg–larval, five larval and two larval–pupal parasitoids. The two most abundant parasitoids in both countries were two Braconidae: the egg-larval parasitoid Chelonus bifoveolatus and the larval parasitoid Coccygidum luteum. Parasitism rates were determined in three Ghanaian regions and averages varied from 0% to 75% between sites and from 5% to 38% between regions. These data provide an important baseline for the development of various biological control options. The two egg parasitoids, Telenomus remus and Trichogramma sp. can be used in augmentative biological control and investigations should be conducted to assess how cultural practices can enhance the action of the main parasitoids, C. luteum and Ch. bifoveolatus, in the field. Understanding the parasitoid complex of S. frugiperda in Africa is also necessary before any development of classical biological controls involving the introduction of parasitoids from the Americas.

Novel species of fungi described in this study include those from various countries as follows: Antarctica , Apenidiella antarctica from permafrost, Cladosporium fildesense fromanunidentifiedmarinesponge. Argentina , Geastrum wrightii onhumusinmixedforest. Australia , Golovinomyces glandulariae on Glandularia aristigera, Neoanungitea eucalyptorum on leaves of Eucalyptus grandis, Teratosphaeria corymbiicola on leaves of Corymbia ficifolia, Xylaria eucalypti on leaves of Eucalyptus radiata. Brazil, Bovista psammophila on soil, Fusarium awaxy on rotten stalks of Zea mays, Geastrum lanuginosum on leaf litter covered soil, Hermetothecium mikaniae-micranthae (incl. Hermetothecium gen. nov.)on Mikania micrantha, Penicillium reconvexovelosoi in soil, Stagonosporopsis vannaccii from pod of Glycine max. British Virgin Isles , Lactifluus guanensis onsoil. Canada , Sorocybe oblongispora on resin of Picea rubens. Chile, Colletotrichum roseum on leaves of Lapageria rosea. China, Setophoma caverna fromcarbonatiteinKarstcave. Colombia , Lareunionomyces eucalypticola on leaves of Eucalyptus grandis. Costa Rica, Psathyrella pivae onwood. Cyprus , Clavulina iris oncalcareoussubstrate. France , Chromosera ambigua and Clavulina iris var. occidentalis onsoil. French West Indies , Helminthosphaeria hispidissima ondeadwood. Guatemala , Talaromyces guatemalensis insoil. Malaysia , Neotracylla pini (incl. Tracyllales ord. nov. and Neotra- cylla gen. nov.)and Vermiculariopsiella pini on needles of Pinus tecunumanii. New Zealand, Neoconiothyrium viticola on stems of Vitis vinifera, Parafenestella pittospori on Pittosporum tenuifolium, Pilidium novae-zelandiae on Phoenix sp. Pakistan , Russula quercus-floribundae onforestfloor. Portugal , Trichoderma aestuarinum from salinewater. Russia , Pluteus liliputianus on fallen branch of deciduous tree, Pluteus spurius on decaying deciduouswoodorsoil. South Africa , Alloconiothyrium encephalarti, Phyllosticta encephalarticola and Neothyrostroma encephalarti (incl. Neothyrostroma gen. nov.)onleavesof Encephalartos sp., Chalara eucalypticola on leaf spots of Eucalyptus grandis_× urophylla, Clypeosphaeria oleae on leaves of Olea capensis, Cylindrocladiella postalofficium on leaf litter of Sideroxylon inerme , Cylindromonium eugeniicola (incl. Cylindromonium gen. nov.)onleaflitterof Eugenia capensis , Cyphellophora goniomatis on leaves of Gonioma kamassi , Nothodactylaria nephrolepidis (incl. Nothodactylaria gen. nov. and Nothodactylariaceae fam. nov.)onleavesof Nephrolepis exaltata , Falcocladium eucalypti and Gyrothrix eucalypti on leaves of Eucalyptus sp., Gyrothrix oleae on leaves of Olea capensis subsp. macrocarpa , Harzia metro sideri on leaf litter of Metrosideros sp., Hippopotamyces phragmitis (incl. Hippopota- myces gen. nov.)onleavesof Phragmites australis , Lectera philenopterae on Philenoptera violacea , Leptosillia mayteni on leaves of Maytenus heterophylla , Lithohypha aloicola and Neoplatysporoides aloes on leaves of Aloe sp., Millesimomyces rhoicissi (incl. Millesimomyces gen. nov.) on leaves of Rhoicissus digitata , Neodevriesia strelitziicola on leaf litter of Strelitzia nicolai , Neokirramyces syzygii (incl. Neokirramyces gen. nov.)onleafspotsof

Key chili and maize growing areas of Pakistan were selected for a focused baseline study of the levels of Aspergillus spp. Investigations were undertaken using a combination of molecular and culture-based techniques. Samples investigated included soil samples, one-year-old corn cobs, and fresh chili from selected locations. Aspergillus strains obtained from corn cobs were screened using coconut milk agar, resulting in one strain that was positive for aflatoxin production. Whole genome sequencing (WGS) with low coverage techniques were employed to screen the isolates for differences in the ribosomal RNA gene cluster and mitochondrial genome, with the aflatoxigenic strain proving to have a distinctive profile. Finally, strains were subjected to matrix-assisted laser-desorption and ionization time-of-flight mass spectrometry (MALDI-ToF-MS) in order to obtain a proteomic ‘fingerprint’ which was used to distinguish the aflatoxigenic strain from the other isolates. The next generation sequencing (NGS) study was broadened to incorporate metabarcoding with ITS rRNA for determining the microbial biodiversity of the soil samples and presumptive screening for the presence of aflatoxigenic strains. Using information gleaned from the WGS results, a putative aflatoxigenic operational taxonomic unit (OTU) was observed in four of the 15 soil samples screened by metabarcoding. This method may have beneficial applications in early detection and surveillance programs in agricultural soils and commodities.

Lupins anthracnose is a destructive seed and airborne disease caused by Colletotrichum lupini, affecting stems and pods. Primary seed infections as low as 0.01–0.1% can cause very severe yield losses. One of the most effective management strategies is the development of a robust and sensitive seed detection assay to screen seed lots before planting. PCR-based detection systems exhibit higher levels of sensitivity than conventional techniques, but when applied to seed tests they require the extraction of PCR-quality DNA from target organisms in backgrounds of saprophytic organisms and inhibitory seed-derived compounds. To overcome these limitations, a new detection protocol for C. lupini based on a biological enrichment step followed by a PCR assay was developed. Several enrichment protocols were compared with Yeast Malt Broth amended with ampicillin, streptomycin, and lactic acid were the most efficient. A species-specific C. lupini primer pair was developed based on rDNA IGS sequences. The specificity was evaluated against 17 strains of C. lupini, 23 different Colletotrichum species, and 21 different organisms isolated from seeds of Lupinus albus cv. Multitalia, L. luteus cv. Mister, and L. angustifolius cv. Tango. The protocol described here enabled the detection of C. lupini in samples artificially infected with less than 1/10,000 infected seed.