Phomopsis husk rot of macadamia in Australia and South Africa caused by novel Diaporthe species

Phomopsis husk rot (PHR) in macadamia is a disease of economic importance in major commercial production areas in Australia and South Africa. Effective control of PHR is hindered by limited knowledge about its aetiology and epidemiology. The diversity and pathogenicity of more than 50 isolates of Diaporthe associated with PHR in macadamia orchards in Australia and South Africa was assessed. Multilocus phylogenetic analyses of DNA sequences of the ITS, tef1α, and tub2 gene loci revealed four novel clades that are described as Diaporthe australiana sp. nov., D. drenthii sp. nov., D. macadamiae sp. nov., and D. searlei sp. nov. Pathogenicity tests with representative isolates found that all four species caused PHR of varying severity between and within species, as well as between the two macadamia cultivars HAES 344 and HAES 816. The Australian species, D. australiana, was the most aggressive species compared with the three South African species. This study improves our understanding of the aetiology of PHR in macadamia and paves the way for more effective disease management.     

A serious canker disease caused by Immersiporthe knoxdaviesiana gen. et sp. nov. (Cryphonectriaceae) on native Rapanea melanophloeos in South Africa

Recent disease surveys in the Western Cape province of South Africa have revealed a previously unknown and serious stem canker disease on native Rapanea melanophloeos (Myrsinaceae, Ericales) trees. Cankers commonly result in the death of branches or entire stems. Fruiting structures typical of fungi in the Cryphonectriaceae were observed on the surfaces of cankers. In this study, the fungus was identified and its pathogenicity to R. melanophloeos was tested. Multigene phylogenetic analyses based on DNA sequences of the partial LSU gene, ITS region of the nuclear ribosomal DNA gene and two regions of the β‐tubulin (BT) gene, showed that the fungus represents a formerly undescribed genus and species in the Cryphonectriaceae. The fungus was also morphologically distinct from other genera in this family. Inoculation trials showed that the fungus described here as Immersiporthe knoxdaviesiana gen. et sp. nov. is an aggressive pathogen of R. melanophloeos trees.     

Corticimorbus sinomyrti gen. et sp. nov. (Cryphonectriaceae) pathogenic to native Rhodomyrtus tomentosa (Myrtaceae) in South China

The Cryphonectriaceae includes many important tree pathogens, especially of the Myrtales. Disease surveys on Myrtales in South China revealed a stem canker disease on native Rhodomyrtus tomentosa (Myrtaceae, Myrtales) trees in the proximity of Eucalyptus plantations in GuangXi Province and in a natural forestry area in the Hong Kong Region. Fruiting structures with typical characteristics of the Cryphonectriaceae were observed on the canker surfaces. The fungus was identified based on DNA sequence comparisons and morphological features, and its pathogenicity was tested on R. tomentosa under field conditions. DNA sequence comparisons for the partial large subunit, partial internal transcribed spacer nuclear ribosomal DNA, and two regions of the β‐tubulin gene showed that the fungus represents a previously undescribed genus and species in the Cryphonectriaceae. Based on morphology, the fungus is most similar to species in the genus Chrysoporthe, but can be distinguished from this and the other genera of Cryphonectriaceae. The fungus provided with the name Corticimorbus sinomyrti gen. et sp. nov., is described. Field inoculations showed that it is highly pathogenic to R. tomentosa trees, with the ability to kill inoculated branches within 4 weeks. Pathogenicity tests also showed that C. sinomyrti is pathogenic to a tested Eucalyptus clone. Pathogenicity on both native R. tomentosa and non‐native Eucalyptus suggests that this fungus should be monitored carefully to limit its possible spread to commercially grown Eucalyptus in South China.     

Anthracnose of Sansevieria trifasciata caused by Colletotrichum sansevieriae sp. nov.

A Colletotrichum sp. was isolated from water-soaked lesions on sansevieria (Sansevieria trifasciata Prain cv. Laurentii) in Japan. Classifying the species only from the morphology of the fungus was difficult; therefore, host range was tested and the ribosomal DNA ITS2 region was phylogenetically analyzed. The fungus was pathogenic only on sansevieria among 20 test plants belonging to 11 families. In a phylogenetic analysis with the neighbor-joining method, the two isolates used formed a single-isolate clade. The fungus is thus proposed to be a new species, Colletotrichum sansevieriae. This report is the first of anthracnose on sansevieria.     

Pewenomyces kutranfy gen. nov. et sp. nov. causal agent of an important canker disease on Araucaria araucana in Chile

Araucaria araucana, (commonly referred to as araucaria, pewen, or monkey puzzle tree) is an ancient conifer endemic to the Chilean and Argentinian mountain ranges where it has a sacred relevance to indigenous communities. During 2015, a serious disease was noticed on trees of all ages in most of the natural distribution of this iconic tree. Four areas were surveyed, and the most important symptoms of the disease were cankers on branches and stems resulting in copious resin exudation. Trees were monitored for a period of two years and isolations were made from the cankers. Field observations showed that the disease typically begins on the leaves or at the leaf bases and progresses downwards to initiate cankers that can girdle branches or stems within a two-year period. Black ascomata, resembling those of Caliciopsis species previously described from A. araucana, were consistently found developing in the cankers from which isolations were made. Phylogenetic analyses of the ITS, nucSSU, and nucLSU gene regions showed that the fungus resides in the Coryneliaceae but is distinct from other genera in that family. The morphological characteristics and phylogenetic position of the fungus show that it represents a new genus and species, described here as Pewenomyces kutranfy gen. nov. et sp. nov. Pathogenicity trials on trees under field conditions confirmed that this newly described fungus is able to cause cankers on A. araucana similar to those found under natural conditions.     

A new anthracnose disease of pyrethrum caused by Colletotrichum tanaceti sp. nov

A new pathogen of pyrethrum (Tanacetum cinerariifolium) causing anthracnose was described as Colletotrichum tanaceti based on morphological characteristics and a four‐gene phylogeny consisting of rDNA‐ITS, β‐tubulin (TUB2), glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) and actin (ACT) gene sequences. The fungus produced perithecia in culture, requiring an opposite mating type isolate in a heterothallic manner. The initial infection strategy on pyrethrum leaves involved the formation of appressoria followed by production of multilobed infection vesicles in the epidermal cells. Infection and colonization then proceeded through thinner secondary hyphae, which resulted in the initial production of water‐soaked lesions followed by black necrotic lesions. The infection process was suggestive of a hemibiotrophic infection strategy. Moreover, phylogenetic analysis clearly showed that C. destructivum, C. higginsianum and C. panacicola were separate species that also had similar intracellular hemibiotrophic infection strategies as C. tanaceti, which all clustered in the C. destructivum complex. Colletotrichum spp. were detected at 1% incidence in seed of 1 of 19 seed lines, indicating the potential for seed as a source of inoculum into crops. Colletotrichum tanaceti was detected in leaf lesions from 11 of 24 pyrethrum fields surveyed between April and July 2012, at a frequency of 1·3–25·0% of lesions. Anthracnose probably contributes to the complex of foliar diseases reducing green leaf area in pyrethrum fields in Australia.     

Tan spot: a new disease of pyrethrum caused by Microsphaeropsis tanaceti sp. nov.

The isolation frequency of Microsphaeropsis sp. in spring in association with necrotic lesions on leaves in Tasmanian pyrethrum (Tanacetum cinerariifolium) fields has increased substantially since first identification in 2001. Examination of morphological features and sequencing of the internal transcribed spacer region (ITS) resulted in the identification of a new species, herein described as Microsphaeropsis tanaceti sp. nov. The pathogenicity of three M. tanaceti isolates to two pyrethrum cultivars was confirmed by inoculating glasshouse‐grown plants in three experiments. No significant differences in the susceptibility of the two cultivars to infection by M. tanaceti were found. Symptoms were tan‐coloured spots which coalesced around the margins of the leaves. Therefore, the name ‘tan spot’ is proposed for this new disease of pyrethrum. The sensitivity of seven M. tanaceti isolates to difenoconazole and azoxystrobin, commonly used fungicides for the management of foliar diseases in spring, was assessed under in vitro conditions. Sensitivity testing for difenoconazole was conducted using a mycelial growth assay on potato dextrose agar, whilst testing for sensitivity to azoxystrobin used a conidial germination assay on water agar. Microsphaeropsis tanaceti was found to be more sensitive to azoxystrobin than difenoconazole, with complete inhibition of conidial germination at concentrations above 0·625 µg a.i. mL^?1. By comparison, concentrations of 50 µg a.i. difenoconazole mL^?1 or greater were required for significant inhibition of mycelial growth. It therefore appears likely that there is currently some control of tan spot as a result of the use of azoxystrobin and to a lesser extent, difenoconazole, for the control of other diseases.     

The morphology,behaviour and molecular phylogeny of Phytophthora taxon Salixsoil and its redesignation as Phytophthora lacustris sp. nov.

Since its first isolation from Salix roots in 1972, isolates of a sexually sterile Phytophthora species have been obtained frequently from wet or riparian habitats worldwide and have also been isolated from roots of Alnus and Prunus spp. Although originally assigned to Phytophthora gonapodyides on morphological grounds, it was recognized that these isolates, informally named P. taxon Salixsoil, might represent a separate lineage within ITS Clade 6. Based on phylogenetic analyses and comparisons of morphology, growth‐temperature relationships and pathogenicity, this taxon is formally described here as Phytophthora lacustris sp. nov. Isolates of P. lacustris form a clearly resolved cluster in both ITS and mitochondrial cox1 phylogenies, basal to most other Clade 6 taxa. Phytophthora lacustris shares several unusual behavioural properties with other aquatic Clade 6 species, such as sexual sterility and tolerance of high temperatures, that have been suggested as adaptations to riparian conditions. It appears to be widespread in Europe and has also been detected in Australia, New Zealand and the USA. It was shown to be weakly or moderately aggressive on inoculation to Alnus, Prunus and Salix. The extent of P. lacustris’ activity as a saprotroph in plant debris in water and as an opportunistic pathogen in riparian habitats needs further investigation. Its pathogenic potential to cultivated fruit trees also deserves attention because P. lacustris has apparently been introduced into the nursery trade.     

Phytophthora andina sp. nov., a newly identified heterothallic pathogen of solanaceous hosts in the Andean highlands

A blight disease on fruits and foliage of wild and cultivated Solanum spp. was found to be associated with a new species of Phytophthora. The proposed novel species is named Phytophthora andina Adler & Flier, sp. nov. based on morphological characteristics, pathogenicity assays, mitochondrial DNA haplotyping, AFLP fingerprinting and nuclear and mitochondrial DNA sequence analyses. Isolates of P. andina (n = 48) from the Andean highland tropics of Ecuador were collected from 1995 to 2006. Phytophothora andina is closely related to P. infestans and has semipapillate, ellipsoidal sporangia borne on sympodially branched sporangiophores. It is heterothallic and produces amphigynous antheridia. The species consists of several clonal lineages, including the EC‐2 and EC‐3 RFLP lineages, which were described previously as P. infestans. Approximately 75% of isolates react as compatibility type A2 when paired with an A1 compatibility type isolate of P. infestans. However, when A2 isolates from the Anarrhichomenum section of Solanum were paired in all combinations, viable oospores were obtained in several crosses, suggesting that there is a unique compatibility interaction in P. andina that is complementary to that described in P. infestans. Nuclear and mitochondrial sequence analysis supported the species designation of P. andina. This newly identified heterothallic pathogen shares a common ancestor with P. infestans and may have arisen from hybridization events with sister taxa in the Andes.     

Fruit spot of sweet lime (<Emphasis Type="Italic">Citrus limetta</Emphasis>) caused by <Emphasis Type="Italic">Septoria</Emphasis> sp. in Peru

In 2002, a severe fruit spot of sweet lime (Citrus limetta) was observed in Piura and Lambayeque provinces in northern Peru. Affected fruits showed large oval and sunken lesions, often surrounded by chlorotic haloes. Septoria sp. was isolated from affected fruits. Sweet lime isolates showed larger pycnidia and pycnidiospores than those of Septoria spp. previously described on citrus. In addition, phylogenetic analysis of the ITS sequences clearly separated the sweet lime isolates from S. citri and S. citricola. Isolates were pathogenic to detached sweet lime fruits and the fungus was isolated from lesions on inoculated fruits.     

葡萄座腔菌属（Botryosphaeria）引起的果树病害及研究进展

葡萄座腔菌属（Botryosphaeria）真菌在国内外分布广泛,是许多果树常见的病原菌、内生菌和腐生菌,能引起果实腐烂、叶斑、坏死、溃疡和流胶等症状。本文概述了该属真菌引起的果树病害的发生、危害症状、主要病原菌种类、流行学和病害控制、栽培管理策略;展望今后的研究方向。     

Extracellular ATP functions in alleviating the decrease of PSII photochemistry caused by the infection of Xanthomonas campestris pv. phaseoli

Extracellular ATP (eATP) can function as a signalling molecule to regulate a wide range of cellular processes. This present work investigated the role of eATP in mediating the change of PSII (photosystem II) photochemistry of the tissues of Phaseolus vulgaris leaves infected with Xanthomonas campestris pv. phaseoli (Xcp). Infection of the leaves with Xcp caused a significant decrease in the the PSII maximal photochemical efficiency (F_v/F_m), the maximum quantum efficiency of PSII photochemistry at illumination (F_v′/F_m′), the PSII operating efficiency (Φ_PSII), the rate of non‐cyclic electron transport through PSII (ETR), photochemical quenching (qP), and eATP level in the tissues of the infected leaves. At the same time, the levels of non‐photochemical quenching (qN) and the quantum yield of regulated energy dissipation of PSII (Y(NPQ)) were significantly increased. Application of exogenous ATP at 0·2 mm to uninfected leaves had no significant effect on any of the chlorophyll fluorescence parameters being measured. However, in the tissues of infected leaves, the application of exogenous ATP alleviated the decreases of the F_v/F_m, F_v′/F_m′, Φ_PSII, ETR, qP and eATP level, and also abolished the increases of qN and Y(NPQ). These results suggest that the change of PSII photochemistry by pathogen infection could be mediated by eATP.     

Phytophthora acerina sp. nov., a new species causing bleeding cankers and dieback of Acer pseudoplatanus trees in planted forests in northern Italy

A severe dieback of Acer pseudoplatanus trees was noticed in planted forest stands in northern Italy in 2010. Affected trees showed collar rot and aerial bleeding cankers along the stems, leading to crown dieback and eventually death. An unknown Phytophthora species was consistently isolated from necrotic bark and xylem tissue and from rhizosphere soil. Based on its unique combination of morphological and physiological characters and phylogenetic analysis, this new taxon is here described as Phytophthora acerina sp. nov. Phylogenetic analysis of ITS, cox1 and β‐tubulin gene regions demonstrated that P. acerina is unique and forms a separate cluster within the ‘P. citricola complex’, closely related to P. plurivora. Phytophthora acerina is homothallic with smooth‐walled oogonia, thick‐walled, mostly aplerotic oospores with a high abortion rate, paragynous antheridia, and persistent, morphologically variable semipapillate sporangia. Four to 5‐week‐old cultures produced globose to subglobose, appressoria‐like and coralloid hyphal swellings and characteristic stromata‐like hyphal aggregations. Optimum and maximum temperatures for growth are 25°C and 32°C, respectively. Genetic uniformity of all 15 studied isolates and the apparent absence of this species in the extensive surveys of nurseries, forests and seminatural ecosystems conducted in the previous two decades across Europe indicate a recent clonal introduction to northern Italy. Under‐bark inoculation tests demonstrated high aggressiveness of P. acerina to A. pseudoplatanus indicating that this pathogen might be a serious risk to maple plantations and forests in Europe.     

Quantification of Cylindrocarpon destructans f. sp. panacis in soils by real-time PCR

Ginseng ( Panax quinquefolius ) is an important cash crop in various regions of North America, but yields are often reduced by various root pathogens. A quantitative real-time PCR (qPCR) assay for Cylindrocarpon destructans f. sp. panacis (CDP), the cause of a root rot and replant disease which discourages successive cropping of ginseng on the same site, was developed to quantify the levels of this pathogen in soils previously cropped with ginseng. DNA was extracted from 5-g samples of soil. In pasteurized soils which were re-infested with varying levels of the pathogen, qPCR estimates of pathogen DNA were significantly correlated with disease severity ( r  = 0·494) and with counts of colony-forming units ( r  = 0·620) obtained with an agar medium. In several naturally infested field soils, qPCR estimates of CDP-DNA concentration were significantly correlated with disease severity ( r  = 0·765) and these concentrations were estimated to range from 0 to 1·48 ng g⁻¹ dried soil. A principal components analysis did not show any strong relationships between soil chemistry factors and the concentration of pathogen DNA. The approach outlined here allows the quantification of current populations of CDP in soil many years after ginseng cultivation and the prediction of disease severity in future crops. The method should be generally applicable to root diseases of many crops.     

Molecular characterisation of vegetative compatibility groups in Fusarium oxysporum f. sp. radicis-lycopersici and f. sp. lycopersici by random amplification of polymorphic DNA and microsatellite-primed PCR

Random amplification of polymorphic DNA (RAPD-PCR) analysis was conducted on 48 isolates of Fusarium oxysporum f. sp. radicis-lycopersici (F.o.r.l.) from different geographic regions, representing all known vegetative compatibility groups (VCGs) except VCG 0097 and VCG 0099 and on eight isolates of F.oxysporum f. sp. lycopersici (F.o.l.), representing VCGs 0030, 0031, 0032 and 0033. Upon UPGMA (unweighted pair-group method with arithmetic averages) analysis of 86 RAPD-PCR markers generated by 16 informative primers and 44 markers obtained with eight microsatellite primers, a close relatedness was evident for F.o.r.l. isolates in VCGs 0090, 0092, 0096, and, to a lesser extent, for those in VCG 0093. Representatives of VCG 0091 formed a distinct group, while F.o.r.l. isolates in VCGs 0094 and 0098 were not distinguishable by the tested markers, most of which were also shared by F.o.l. isolates belonging to VCGs 0031 and 0033. F.o.l. isolates in VCGs 0030 and 0032 shared most of the molecular markers. The correlation between RAPD-PCR and microsatellite genetic distance was highly significant (R² = 0.77; P by Mantel test < 0.001). The molecular variability observed in both formae speciales is discussed in relation to the development of F.o.r.l.- and F.o.l.-specific diagnostic tools.     

Variation in susceptibility of lettuce cultivars to fusarium wilt caused by Fusarium oxysporum f.sp. lactucae

Lettuce cultivars adapted to Californian growing conditions were screened for resistance to fusarium wilt caused by Fusarium oxysporum f.sp. lactucae in order to determine if differences in susceptibility among currently grown cultivars might contribute to management of this disease. Based on a preliminary evaluation of 46 cultivars, eight that were among the most resistant of their horticultural type (iceberg, romaine or leaf) were selected for further testing. The relative susceptibility of these cultivars was assessed by: (i) root-dip inoculation, (ii) sowing seeds into infested potting mix and (iii) transplanting seedlings into an infested field. Evaluations of disease severity showed that both methods (i) and (ii) produced cultivar rankings that were significantly correlated with rankings from field trials [method (iii)]. Two romaine and two leaf cultivars were highly resistant to fusarium wilt (mean disease severity rating of ≤1·3 on a 1–4 scale) under all test conditions. Other romaine and leaf cultivars, however, were highly susceptible in root–dip tests, so there was no consistent association between cultivar type and susceptibility to fusarium wilt. Likewise, there was considerable variation in susceptibility to wilt among iceberg cultivars, but all were significantly more susceptible than the most resistant romaine and leaf cultivars.