Identification of Berkeleyomyces rouxiae causing black root rot disease on cotton seedlings in New South Wales,Australia

Black root rot disease of cotton seedlings caused by Thielavioposis basicola was first reported in New South Wales (NSW), Australia in 1990. In 2018, T. basicola was reclassified into a new genus Berkeleyomyces, accommodating two closely related species: B. basicola and B. rouxiae. However, species status of cotton-T. basicola in NSW remains unsolved. Ninety-five isolates recovered from black root rot diseased cotton seedlings sampled across NSW in 2017/18 season was subjected to morphological, multigene sequencing (ITS, MCM7, RPB2), and pathogenicity assessments for their species identification. Berkeleyomyces rouxiae was accurately identified as the causal agent of black root rot of cotton.

     

Morphological and molecular characterization of Phytophthora species associated with root and crown rot of pomegranate in Iran

The purpose of this research was to identify the pathogens causing root and crown rot in major pomegranate-growing areas of Iran. Infected tissue samples were collected from trees with symptoms from 49 pomegranate orchards in four provinces of Iran: Fars, Markazi, Isfahan, and Kohgiluyeh va Boyer-Ahmad. In total, 23 Phytophthora spp. isolates were obtained, which were identified as P. cryptogea species complex (12 isolates) and P. cinnamomi (11 isolates) based on morphological characters. Molecular confirmation of identification was performed by inference of phylogeny of ITS-rDNA regions, β-tubulin gene, and the mitochondrial gene cytochrome c oxidase subunit 1. The results of our phylogenetic analysis confirmed the morphological identification of P. cinnamomi isolates and placed them in Clade 7c of Phytophthora. In addition, the P. cryptogea species complex isolates, despite morphological similarities, were in fact four distinct species including P. cryptogea sensu stricto (two isolates), P. pseudocryptogea (one isolate), P. sp. kelmania (one isolate), and P. erythroseptica (six isolates). This is the first report of pomegranate root and crown rot caused by P. cinnamomi and P. cryptogea species complex.     

rDNA-based characterization of a new binucleate Rhizoctonia spp. causing root rot on kale in Brazil

In this paper we present the first report of the occurrence of a binucleate Rhizoctonia spp. causing hypocotyl and root rot in kale in Brazil. Rhizoctonia spp. were isolated from kale (Brassica oleracea var. acephala) with symptoms of hypocotyl and root rot. The isolates, characterized as binucleate Rhizoctonia spp., did not show an anastomosis reaction with any of the binucleate Rhizoctonia spp. testers used. The pathogenicity of the isolates was tested under greenhouse conditions; all isolates were pathogenic and showed different symptom severities on kale. The ITS-5.8S rDNA sequences of kale isolates and 50 testers (25 binucleate Rhizoctonia spp. and 25 Rhizoctonia solani) were compared in order to characterize the genetic identity of Rhizoctonia spp. infecting kale. The kale isolates showed genetic identities ranging from 99.3 to 99.8% and were phylogenetically closely related to CAG 7 (AF354084), with identities of 98.5 and 98.7%. It is suggested that the binucleate Rhizoctonia spp. causing hypocotyl and root rot on kale Brazil comprises a new AG not yet described.     

A novel lineage in the Fusarium incarnatum-equiseti species complex is one of the causal agents of fusarium rot on melon fruits in Northeast Brazil

Fusarium rot on melon fruits is a postharvest disease and its importance in Brazil is increasing since its first report in 1999. Initially this disease was attributed to the fungus Fusarium semitectum based on pathogen morphology. However, there is controversy regarding the aetiology of this disease because, in the current species concept based on phylogenetic analysis, F. semitectum is regarded as a synonym to different Fusarium species. With the objective of elucidating the disease aetiology in Northeast Brazil, Fusarium isolates were obtained from melon fruits showing rot symptoms in the main producing areas of the country. From the phylogenetic analyses of TEF1 and RPB2 gene sequences, the isolates were identified as belonging to two phylogenetic species of the Fusarium incarnatum-equiseti species complex (FIESC), one in the Incarnatum clade (Fusarium sulawense) and the other in the Equiseti clade, which corresponds to a new lineage. The newly identified lineage is close to Fusarium lacertarum. Isolates from the two species showed morphological characteristics typical of the Incarnatum and Equiseti clades, agreeing with the molecular identification, and were pathogenic when inoculated on melon fruits. This is the first report of F. sulawense on melon fruits. The data generated in this study are potentially useful for a better management of the fusarium rot on melon.     

PCR‐based identification of Pythium spp. causing cavity spot in carrots and sensitive detection in soil samples

On the basis of ITS sequences PCR primers were designed for the identification of the five Pythium species found to be most important for the development of carrot cavity spot in Norway: P. intermedium, P. sulcatum, P. sylvaticum, P. violae and P. ‘vipa’. The P. ‘vipa’ isolates had a unique ITS sequence, differed morphologically from all other Pythium isolates, and thus probably represent a new species. The PCR primers were species‐specific with no cross‐reaction to other Pythium species or to fungal isolates from carrot tested. The detection limits varied for the different primer pairs. The two most sensitive assays allowed detection of as little as 5 fg DNA. All five Pythium species could be detected in lesions from diseased carrots. Weak positive signals were obtained from some carrot samples without symptoms. PCR assays allowed detection of pathogens in soil. In samples of soil known to produce cavity spots on cropped carrots, strong signals were obtained. In several soil samples more than one of the five Pythium species could be detected. The utilization of this diagnostic PCR assay in analysis of field soil and carrot tissue might in the future be exploited to reduce the incidence of this serious carrot disease.     

Diversity and pathogenicity of Fusarium graminearum species complex from maize stalk and ear rot strains in northeast China

The Fusarium graminearum species complex (FGSC) is an important group of pathogens distributed in maize‐producing areas worldwide. This study investigated the genetic diversity and pathogenicity of 40 FGSC isolates obtained from stalk rot and ear rot samples collected from 42 locations in northeastern China during 2013 and 2014. A phylogenetic tree of translation elongation factor (EF‐la) sequences designated the 40 isolates as F. graminearum sensu stricto (67.5%) and F. boothii (32.5%). By using inter‐simple sequence repeat analysis (ISSR), it was shown that the isolates were divided into two clades, which corresponded to the species identity of the isolates. However, the isolates from the two different diseases could not be distinguished in pathogenicity. The disease severity index of seedlings inoculated with stalk isolates was slightly higher than that of seedlings inoculated with isolates from infected ears, whereas the pathogenicity of the stalk and ear isolates were identical.     

Characterization of isolates of binucleate Rhizoctonia spp. associated with strawberry black root rot complex using fatty acid methyl ester (FAME) profiles

Black root rot is an important disease of strawberry caused by a complex of fungi that includes species of Rhizoctonia. In this study, a modified MIDI method (Microbial Identification System) was investigated for its utility to differentiate isolates of the three different anastomosis groups (AGs) of binucleate Rhizoctonia spp., associated with strawberry black root rot complex representing AG-A, AG-G, and AG-I. A total of 11 fatty acids were detected, and the FAME profiles for isolates of the three different AGs of Rhizoctonia spp. varied quantitatively and qualitatively. Moreover, the modified MIDI method will be a useful discriminatory tool for fungal identification and classification of the AGs of binucleate Rhizoctonia spp. associated with strawberry black root rot complex.     

Pathogenic fungi involved in root rot of peas in the Netherlands and their physiological specialization

Research on root rot pathogens of peas in the Netherlands has confirmed the prevalence ofFusarium solani, F. oxysporum, Pythium spp.,Mycosphaerella pinodes andPhoma medicaginis var.pinodella. Aphanomyces euteiches andThielaviopsis basicola were identified for the first time as pea pathogens in the Netherlands. Other pathogens such asRhizoctonia solani andCylindrocarpon destructans were also found on diseased parts of roots. F. solani existed in different degrees of pathogenicity, and was sometimes highly specific to pea, dwarf bean of field bean, depending on the cropping history of the field.A. euteiches was specific to peas, whereasT. basicola showed some degree of physiological specialization.     

Colletotrichum falcatum and Fusarium species induce symptoms of red rot in sugarcane in Brazil

Red rot is an important disease of sugarcane, reported from the main producing countries of the crop. The main causal agent is Colletotrichum falcatum, which induces reddish internal stalk rot, resulting in loss of quality and quantity of sugars. While the occurrence of this disease has been reported in plantations in Brazil, the aetiology of the disease is not yet fully understood. We isolated and identified the fungi associated with sugarcane plants showing symptoms of red rot. Thirty isolates were recovered and grouped by morphology within the genera Colletotrichum and Fusarium. Based on phylogenetic analyses of DNA sequences from ITS, ACT, SOD, GAPDH, and EF-1α, eight isolates were identified as C. falcatum (n = 6) and C. plurivorum (n = 2). Twenty-two isolates belonged to the Fusarium fujikuroi species complex and were identified as F. sacchari (n = 8), F. proliferatum (n = 3), and F. madaense (n = 11). Coinfection with C. falcatum and Fusarium species were frequently observed. C. falcatum and Fusarium strains induced reddening of internal tissues and stalk rot in sugarcane plants, symptoms of red rot, while Fusarium species also induced symptoms of pokkah boeng. C. plurivorum, whose sexual stage was observed on the surface of stems, did not induce stalk rot or leaf symptoms. The results obtained in this study clarify the aetiological agents of red rot of sugarcane in Brazil. The conditions that lead to the development of specific symptoms of red rot or pokkah boeng are under investigation.     

Identification of fungal pathogens and analysis of genetic diversity of Fusarium tricinctum causing root rots of alfalfa in north-east China

Alfalfa root rot is a devastating disease complex found worldwide. Population structure and genetic diversity of fungal pathogens causing alfalfa root rot in north-east China are not well understood. In this study, 480 fungal isolates were collected from six major alfalfa-growing regions in Heilongjiang province, China. They were identified as Fusarium tricinctum, F. oxysporum, F. acuminatum, F. solani, F. equiseti, Phoma medicaginis, Plectosphaerella cucumerina, Alternaria alternata, and Chaetomium globosum and caused root rot on alfalfa in greenhouse studies. F. tricinctum was the predominant species among the isolates, and P. medicaginis and C. globosum had not previously been reported causing alfalfa root rot in north-east China. Of the 73 F. tricinctum isolates identified, the majority were moderately or highly aggressive on alfalfa. No isolate of F. tricinctum was sensitive to carbendazim (1 and 10 μg/ml), indicating that, although commonly used, it is not suitable for management of the disease in this area. F. tricinctum isolates were analysed using AFLP markers and divided into eight genetic groups with 28 pairs of primers. Analysis of molecular variance indicated significant correlation between genetic groups of F. tricinctum isolates and their geographical locations or aggressiveness. Pairwise comparison and STRUCTURE analysis also indicated that geographical locations and aggressiveness of isolates had a significant effect on population differentiation. This study provides insight into the genetic diversity and reproductive biology of F. tricinctum, enhances understanding of the population diversity of alfalfa root rot pathogens in north-east China, and facilitates development of effective strategies for managing this destructive disease complex.     

Genetic diversity and population structure of Lasiodiplodia theobromae from different hosts in northeastern Brazil and Mexico

Lasiodiplodia theobromae is one of the most frequent fungal pathogens associated with dieback, gummosis, leaf spot, stem-end rot and fruit rot symptoms in cashew, mango, papaya and grapevine. In this study, the variation in the genetic diversity of 117 L. theobromae isolates from northeastern Brazil (n = 100) and Mexico (n = 17), which were collected from these four crops, was analysed using microsatellite markers. The results revealed low genetic diversity among L. theobromae populations and the existence of two genetic groups. All Mexican isolates were grouped with Brazilian isolates, suggesting a low level of differentiation between these populations. Furthermore, no evident host or climate-based population differentiation was observed for L. theobromae in Brazil. The populations studied were mostly clonal, but additional studies are needed to better understand the mode of reproduction of the pathogen. The low genetic diversity of L. theobromae populations in northeastern Brazil suggests that resistant cultivars could be used as a durable management strategy to reduce the impact of the diseases caused by this pathogen.     

Molecular phylogeny and morphological characterization of the aetiological agent of sour rot on fruits and vegetables in Brazil

In Brazil, sour rot is an important postharvest disease on fruits and vegetables. Geotrichum candidum (synonym Galactomyces candidus) has been reported as the main species causing this disease. However, the identity of the causal agent is still uncertain. This research aimed to determine the identity of 165 fungal isolates associated with sour rot obtained from fruits and vegetables in Brazil, and to evaluate the effect of different temperatures on the incidence of sour rot on artificially inoculated tomato fruits. Based on the phylogenetic analyses of DNA sequences from the D1/D2 domain of the large subunit (LSU) rRNA gene and morphological analyses, 129 samples belonged to Galactomyces candidus, 15 to G. candidum var. citri-aurantii, 6 to G. phurueaense, 2 to Gal. pseudocandidus, 1 to Hyphopichia burtonii, 1 to H. khmerensis, 3 to Saccharomycopsis crataegensis, 1 to S. vini, 1 to Magnusiomyces tetrasperma, 1 to Trichosporon coremiiforme, and 1 to Zygoascus meyerae. Two new species were found, namely, Geotrichum solani (on potato) and Geotrichum spondiadis (on red mombin). All isolates were pathogenic when inoculated on healthy tomato fruits, including the new species of Geotrichum, which were also inoculated into their respective hosts, that is, potatoes and red mombins. To the best of our knowledge, this study is the first to report the presence of five other genera besides Geotrichum associated with sour rot on fruits and vegetables in Brazil, which demonstrates the diversity of fungi and yeasts associated with this disease.     

Thielaviopsis basicola (Berk. et Br.) Ferraris als veroorzaker van een voor Nederland nieuwe ziekte van de tabak

Summary Tobacco black root rot, caused byThielaviopsis basicola (Berk. etBr.) Ferraris has been reported for the first time in the Netherlands. It occurred in tobacco seed beds in the Maas and Waal district, especially in the variety Kentucky Italiana.A part of the literature on this subject is briefly discussed and methods for control are given.     

北京市生菜链格孢根腐病病原菌鉴定

为明确北京市生菜链格孢根腐病的病原菌种类,采用常规组织分离法分离获得病原菌,依据柯赫氏法则对病原菌进行致病力检测,并利用分子生物学技术结合形态学鉴定确定病原菌分类地位。结果显示,从生菜病样组织中分离到2种病原菌共18株,形态学鉴定结果为芸薹链格孢Alternaria brassicae和万寿菊链格孢A. tagetica,分离比例分别为55.6%和44.4%,且二者均能单独侵染生菜根部,前者致病力较后者强,亦能复合侵染。对致病菌株进行GAPDH基因的PCR扩增和测序,并建立了基于GAPDH基因序列的系统发育树,聚类分析结果与形态学鉴定结果一致,因此证实北京市生菜链格孢根腐病是由芸薹链格孢和万寿菊链格孢复合侵染所致。     

Black root rot of cucurbits caused by Phomopsis sclerotioides in Japan and phylogenetic grouping of the pathogen

Although the causal agent of black root rot of Cucurbitaceae in Japan has been proposed as Phomopsis sclerotioides, the species identification of the pathogen has remained inconclusive because of a lack of spore formation. We confirmed that a Japanese isolate of Phomopsis sp. obtained from a diseased pumpkin root produced pycnidia containing α spores in sterilized bean pods. In phylogenetic analyses of rDNA-ITS regions, nine Japanese Phomopsis sp. isolates from melon, watermelon grafted onto bottle gourd, and pumpkin diagnosed with black root rot, formed a single clade with P. sclerotioides standard isolates. We identified the causal agent of the black root rot of melon, pumpkin, bottle gourd, and watermelon in Japan as P. sclerotioides and propose the Japanese name “Phomopsis-negusare-byo” for the disease. Patterns of random amplified polymorphic DNA (RAPD) of these Japanese isolates were also similar to those of P. sclerotioides, thus supporting the species identification. However, mycelial incompatibilities were found for many combinations among these P. sclerotioides isolates, suggesting some genotypic variations of this fungus in Japan at a level that the RAPD analyses cannot discriminate. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB201430 to AB201444     

Ilyonectria palmarum sp. nov. causing dry basal stem rot of Arecaceae

During surveys conducted in 2010–2013, a complete breakage or bending of the trunk and a dry basal stem rot were observed on containerised Brahea armata, B. edulis, Howea forsteriana and Trachycarpus princeps plants in different nurseries located in eastern Sicily (southern Italy). A cylindrocarpon-like species was consistently obtained from diseased palm tissues, while known pathogens of these hosts such as Ganoderma, Phytophthora and Thielaviopsis were not found associated with symptomatic tissues or isolated on standard or selective media. A total of 40 cylindrocarpon-like isolates were collected and characterised based on morphology and DNA phylogeny. Multigene analyses based on the β-tubulin, histone H3, translation elongation factor 1-α, and the internal transcribed spacers (ITS1, 5.8S, ITS2) genes facilitated the identification of a new species, described here as Ilyonectria palmarum. The pathogenicity of one representative isolate collected from each palm species was tested on plants cultivated under nursery conditions and in a growth chamber. All isolates were pathogenic to B. armata, B. edulis, H. forsteriana, and T. princeps and symptoms identical to that observed in nurseries were reproduced. Dry basal stem rot and stem bending caused by Ilyonectria palmarum represents a potentially serious problem for nurseries cultivating containerised palms.     

玉米穗、茎腐病镰孢菌相互关系的血清学研究

运用血清学方法研究了沈阳、河北、北京、吉林4个地区玉米穗、茎腐病镰孢菌在病原学上的相互关系。结果发现:各地区玉米穗、茎腐病串珠镰孢菌(Fusarium moniliforme)具有高度的同源性,亲缘关系近。各地区玉米穗、茎腐病禾谷镰孢菌(F.graminearum)的亲缘关系因地域不同而有差异。沈阳穗、茎腐病禾谷镰孢菌之间差异明显,存在生理分化现象;吉林玉米穗、茎腐病病原物存在较高的相似性,但也有一定的差异;而河北玉米穗、茎腐病禾谷镰孢菌具有高度的同源性。     

Molecular phylogenetic,morphological, and pathogenic analyses reveal a single clonal population of Septoria lycopersici with a narrower host range in Brazil

Septoria leaf spot, caused by Septoria lycopersici, is considered one of the most important diseases of tomato in Brazil. Despite its importance, the disease agent is still poorly studied. Septoria isolates collected from different production regions of Brazil were characterized by molecular, morphological, and pathogenic methods. A set of 104 isolates was sequenced for the DNA Tub, Cal, and EF1-α loci. Ten isolates were selected, according to geographical region of origin and type of leaf lesion (typical or atypical), for morphological characterization and for evaluation of aggressiveness on tomato cultivar Santa Clara. To evaluate the pathogen host range, cultivated and wild Solanaceae plants were inoculated with four selected isolates. The results showed that all isolates grouped with the type isolate of S. lycopersici in maximum likelihood and Bayesian inference trees. The isolates were morphologically similar. All isolates selected for pathogenicity testing on tomato were able to induce typical symptoms of the disease, but differed in their aggressiveness. A total of eight species of Solanaceae were also identified as potential alternative hosts for S. lycopersici. This information will provide a more accurate assessment of the risks involved with the introduction of new crops, especially of the genus Solanum, in areas where the species is already present. In addition, it will provide the basis for the establishment of more efficient methods in the management of Septoria leaf spot of tomatoes in natural conditions and in the different production systems.