First report of Neofusicoccum parvum causing stem and branch canker on Cannabis sativa in Italy

Journal of Plant Diseases and Protection - In 2016, symptoms of stem canker with branches dieback were detected in several Cannabis sativa L. crops, located in the northeast part of Italy (Rovigo...     

Cytospora oleina causing canker and dieback of olive in Greece

Cytospora oleina was isolated from dead twigs of olive associated with old cankers in the Mount Pelion region of central Greece. Inoculation of young trees with pure cultures of the fungus caused twig death in olive, especially when applied in autumn, and xylem discoloration in apple, plum, cherry, peach and apricot.     

核桃枝枯病小新壳梭孢巢式PCR检测方法的建立

近年来,小新壳梭孢Neofusicoccum parvum导致的核桃枝枯病屡有报道,该病防治困难,发病率高,是核桃的一种灾难性病害。为建立N.parvum的快速、灵敏的分子检测体系,根据N.parvum的几丁质合酶1基因(CHS1)及其前后100bp序列,分别设计了两对特异性引物CT-WK3-S/CT-WK3-A和CT-N-S/CT-N-A,建立了N.parvum的巢式PCR检测方法。结果表明,建立的体系可以从不同来源的5个N.parvum菌株中特异性地扩增出97bp的目的条带,灵敏度达30fg/μL,是常规PCR的10倍,而从其近缘种葡萄座腔菌属Botryosphaeriasp.的病原菌及其他供试菌株均未扩增出任何条带。以感染N.parvum的核桃组织总DNA为模板进行检测,可以在发病早期检测出N.parvum的存在。本研究建立的巢式PCR体系可以为核桃枝枯病的田间检测提供思路。     

Botryosphaeriaceae species causing dieback on Annonaceae in Brazil

In Brazil, the Annonaceae species Annona muricata, A. squamosa, A. cherimola and atemoya (a hybrid of A. cherimola and A. squamosa) are cultivated in several regions, and produce fruits that are highly appreciated by consumers and are of great economic importance. Among the several diseases that can affect these crops, dieback is one of the most important, causing damage and, in the most severe cases, death of the plants. Due to the lack of suitable diagnostic studies up to now, this work aimed to identify the Botryosphaeriaceae species that cause dieback on Annonaceae in Brazil. Based on combined phylogenetic analyses of ITS, TEF-1α, TUB2 and RPB2, eight species of Botryosphaeriaceae were identified, namely Lasiodiplodia brasiliense, L. crassispora, L. hormozganensis, L. iraniensis, L. pseudotheobromae, L. subglobosa, L. theobromae and Pseudofusicoccum stromaticum. All species found in this study were pathogenic and caused symptoms of necrosis in stems and dieback. Thus, this study confirms species of Botryosphaeriaceae as causal agents of dieback on Annonaceae in Brazil.     

苹果枝枯病病原菌鉴定及生物学特性

In July 2017, a new shoot dieback disease, caused brown necrotic spots with black granules on the shoots in the late stage, was found in several apple orchards in Yantai, Shandong Province. The fungal pathogen was isolated on PDA plate from the infected shoots using the tissue isolation, which formed white to grey colonies bearing abundant fruiting bodies on Czapek solid media with lactose as carbon source. The purified fungal isolates La1-La3 were obtained from single spore isolation after spore tendrils exuding from pycnidia generated on the plates. This study was to identify and characterize the causal agent . Based on morphological characteristics and analysis of rDNA internal transcribed spacer (ITS), β-tubulin (TUB) and translation elongation factor 1-α (TEF1) nucleotide sequences, the fungal isolates were identified as Lasiodiplodia pseudotheobromae, a new pathogen on apple tree. The test of Koch's rule confirmed that L. pseudotheobromae was the pathogen of this disease. Effects of different temperature, pH value, carbon and nitrogen sources were tested against the growth of isolated fungus in vitro. The results showed that the optimal growth temperature and pH value were at 30℃ and pH 6-9, respectively, while the optimum carbon source was sucrose, and favourable nitrogen sources were sodium nitrate, ammonium nitrate and peptone.     

苹果枝枯病病原菌鉴定及生物学特性

<正>苹果(Malus pumila Mill.)是世界上栽培最为广泛的水果之一,由于其生态适应性强、营养价值高、耐贮性好及供应周期长,相当多的国家都将其列为主要消费果品[1],而我国是世界上苹果种植面积最大、产量最高的国家[2]。我国苹果枝干病害以苹果轮纹病和腐烂病为主,近年来随着气候环境的改变,一些新的病害也逐渐产生。2017年7月对烟台市几个苹果果园调查时,发现一种新病害。该病害主要危害苹果枝条,产生褐色坏死病斑,严重时导致枝条枯死,发     

Production of phytotoxic metabolites by five species of Botryosphaeriaceae causing decline on grapevines, with special interest in the species Neofusicoccum luteum and N. parvum

In recent years an increasing number of species of Botryosphaeriaceae have been associated with grapevine decline worldwide. Five species isolated from declining grapevines in Spain (Botryosphaeria dothidea, Diplodia seriata, Dothiorella viticola, Neofusicoccum luteum and N. parvum) were checked for toxin production in liquid cultures. Cultural conditions for all fungi were adjusted to obtain optimal production of phytotoxic culture filtrates, by growing the fungi in steady liquid cultures of Czapek–Dox broth for different time intervals. Phytotoxicity of D. seriata and N. parvum reached a maximum after 14 days while the remaining species showed the highest phytotoxicity levels after 21 days in culture. All fungi produced hydrophilic high-molecular weight compounds with phytotoxic properties. In addition, N. luteum and N. parvum produced lipophilic low-molecular weight phytotoxins, not detected consistently among the remaining species. This led to a more exhaustive study on the phytotoxicity of N. luteum and N. parvum. Culture filtrates and corresponding extracts of both species were consistently highly phytotoxic in different assays. The gas-chromatography analysis of the acetylated O-methyl glycosides of the phytotoxic exopolysaccharides produced by N. parvum showed these substances to be composed mainly of glucose, mannose and galactose. Results suggest that phytotoxic metabolites could be involved in the virulence of both species in planta.     

Eutypa canker and dieback of almonds

Eutypa canker and dieback of almonds was first recorded in commercial plantations in Greece in mid-1980 and confirmed as due to the ascomycetous fungus Eutypa lata. The disease manifests itself by the formation of cankers at the point of the junction of the branches and the trunk and is associated with pruning wounds made by the growers in order to confront the'non-infectious bud failure'disorder or to remove unwanted limbs close to the crotch. Pathogenicity tests made on young trees of cv. Texas with different isolates originated from grapevine, almond, apricot and pear showed significant differences in virulence. Pathogenicity tests carried out on 20 different almond cultivars gave a significant indication of the existence of tolerant and susceptible cultivars.     

Studies of canker and dieback of oak tree in China,with two Cytospora species described

Oak (Quercus sp.) is an excellent tree species as a windbreak, for water conservation, and for fireproofing in forests in China. However, several trees of this genus were found to be suffering from various fungal diseases. In this study, we evaluated 15 fungal pathogens that can cause dieback and canker disease in oak in China, and discovered two Cytospora species. They were identified as Cytospora quercinum sp. nov. and C. vinacea, based on detailed morphological comparisons and phylogenetic analysis of ITS, LSU, act, rpb2, tef1-α, and tub2 loci. This study is the first record of C. quercinum and C. vinacea as causal agents of dieback in oak based on pathogenicity tests conducted on 2-year-old plants in a greenhouse. In addition, this study also revealed the influence of different conditions on the growth rate of mycelia. Mycelial growth of C. quercinum and C. vinacea occurred at optimum temperatures of 20.1 and 20.8 °C, and optimum pH of 5.4 and 5.3, respectively. For these two species, utilization of glucose and fructose was highly efficient, and sucrose was the least efficient. The habitat of Quercus mongolica indicated that more attention and management are needed to prevent the occurrence of Cytospora disease in the summer in north-eastern China, and in spring and autumn in eastern and northern China. This study contributes to the understanding of the species causing canker or dieback diseases in important economic forest trees, and provides useful information for effective disease management of oak trees in China.     

A presumably exotic Phytophthora species causing dieback of Buxus sempervirens in Germany and Romania

Journal of Plant Diseases and Protection - Roots, collars and rhizosphere soils from diseased boxwood saplings (Buxus sempervirens) from German and Romanian nurseries were shown to be colonized by...     

厚皮香枯梢病病原菌鉴定

厚皮香枯梢病在上海地区发生严重。本研究对上海地区的厚皮香枯梢病进行了病原菌的分离与致病性鉴定,得到其病原菌菌株2017SHTg。根据病原菌的形态特征将其鉴定为小新壳梭孢Neofusicoccum parvum。扩增病原菌2017SHTg的rDNA-ITS序列和翻译延伸因子EF-1α基因序列,进行BLAST序列比对与小新壳梭孢相似性分别为99%和100%;基于rDNA-ITS序列构建系统发育进化树,其在系统发育树上与小新壳梭孢的类群处于同一分支,此结果与形态学鉴定结果一致。     

Potential susceptibility of Australian native plant species to branch dieback and bole canker diseases caused by Phytophthora ramorum

Susceptibility to branch dieback caused by Phytophthora ramorum was tested using a detached branch assay for 66 Australian native plant species sourced from established gardens and arboreta in California. Six of these species were further tested for their susceptibility to bole cankers caused by P. ramorum using a sealed log assay. Isopogon formosus and Eucalyptus denticulata were identified as potentially highly susceptible Australian branch dieback hosts. Thirteen potentially tolerant Australian host species included Banksia attenuata, B. marginata, E. haemastoma, E. regnans, Pittosporum undulatum and Billardiera heterophylla. Eucalyptus regnans was identified as a potentially highly susceptible bole canker host, while E. diversicolor and E. viminalis were considered potentially tolerant species to bole cankers caused by P. ramorum. Phytophthora ramorum was able to infect all 66 species, as confirmed by reisolation. These results extend the known potential host range for P. ramorum, confirm it as a possible threat to Australian plant industries and ecosystems and highlight additional associated hosts that are important in the global horticultural trade, native forests and plantation forestry.     

Susceptibility of grapevine pruning wounds to infection by Lasiodiplodia theobromae and Neofusicoccum parvum

The susceptibility of 1‐ and 2‐year‐old grapevine wood to botryosphaeria canker caused by Lasiodiplodia theobromae and Neofusicoccum parvum was evaluated in California in two seasons. In the 2007 dormant season, pruning‐wound susceptibility was highest when wounds were inoculated immediately after pruning in December (80% of pruning wounds were infected in Chardonnay for both fungal species and 75% and 98% in Cabernet Sauvignon for N. parvum and L. theobromae, respectively). In the 2008 dormant season, pruning‐wound susceptibility was highest in November in Chardonnay (86% and 93% for N. parvum and L. theobromae, respectively) and in December in Cabernet Sauvignon (71% and 75% for N. parvum and L. theobromae, respectively). The lowest infection rates (13–35%) were observed when vines were pruned and inoculated in March in both dormant seasons and for both cultivars. Susceptibility of pruning wounds did not differ significantly (P = 0·7612) between 1‐ and 2‐year‐old wood and consequently, pruning‐wound protection treatments should be applied to all wounds. In conclusion, grapevine pruning wounds were susceptible to infection by L. theobromae and N. parvum to varying extents throughout the dormant season in California (November–March), but, overall, susceptibility of pruning wounds was highest when inoculations were done immediately after pruning and decreased significantly as the interval between pruning and inoculation increased. Results of this study suggest that pruning grapevines in late winter (March) in California would significantly reduce the risk of infection by L. theobromae and N. parvum.     

Biological characterization and genetic diversity analysis of two species of Pestalotiopsis causing twig dieback of Myrica rubra

Chinese bayberry (Myrica rubra Siebold & Zucc) is an evergreen fruit tree with high ecological and economic values in China. In recent years, a new twig dieback disease caused by Pestalotiopsis mangiferae and P. vismiae was observed in major M. rubra-producing areas of Zhejiang and Fujian Provinces, causing serious economic losses. In this study, 16 isolates of P. mangiferae and 27 isolates of P. vismiae were obtained from diseased leaves, roots and branches of M. rubra from different regions. The optimum growth temperature of the two species of Pestalotiopsis was determined to be 20–25 °C, while the optimum temperature for the germination of conidia was 25–35 °C. The two species of Pestalotiopsis showed rich genetic diversity. Inoculating the conidial suspension of one or both of the two species of the Pestalotiopsis on detached leaves or branches of M. rubra could cause lesions surrounding the inoculation sites with the frequency of 100 %. Moreover, necrotic lesions could be observed on inoculated potted plants with the frequencies of 33.3 % for P. mangiferae, 25 % for P. vismiae and 50 % for a mixed inoculum.     

Detection of Botryosphaeriaceae species within grapevine woody tissues by nested PCR, with particular emphasis on the Neofusicoccum parvum/N. ribis complex

Several species of Botryosphaeriaceae and Phaeomoniella chlamydospora are common agents of grapevine decline worldwide. Currently, the use of culture independent PCR based techniques for detection of Botryosphaeriaceae within grapevine tissues has been limited to Botryosphaeria dothidea. In the present study, two Botryosphaeriaceae specific nested PCR assays were developed. One with a narrow target range, to detect Neofusicoccum parvum and the closely related species complex (Neofusicoccum parvum/N. ribis sensu Pavlic et al. Molecular Phylogenetics and Evolution 51:259–268, 2009) and another, with a wider range, to detect all 17 species of Botryosphaeriaceae which have been reported as potential wood pathogens of grapevine. The effectiveness of these assays was validated in vivo on naturally infected wood samples collected from standing vines and dormant grafted rooted cuttings commercialized in Italy by different nurseries in different years. All samples were also screened by means of a previously published nested PCR assay specific for Phaeomoniella chlamydospora. It was found that: 1) propagation material may play an important role as source of primary inoculum, not only of Phaeomoniella chlamydospora, as previously reported, but also for members of the Botryosphaeriaceae, among which Neofusicoccum parvum, Botryosphaeria dothidea and Diplodia seriata are the most common, and 2) multiple infections by different species belonging to Botryosphaeriaceae and/or Phaeomoniella chlamydospora occur frequently both in standing vines and propagation material. This last finding supports the hypothesis that at least some of the non-specific symptoms of grapevine decline may be due to the presence of different pathogens within host tissues.     

桉树溃疡病病原菌的鉴定

 随着桉树人工纯林的面积扩大,桉树病害也日趋加重,严重影响和制约桉树产业的发展。2008年至今,在云南省石林县和红河州桉树种植区发生一种病害,主要为害嫩枝、枝条和主干,发病初期,感病枝条和主干上产生红褐色圆或椭圆形坏死斑,以后逐渐扩大成椭圆或不规则溃疡斑,病变扩展到木质部后,常造成枝条生长畸形,易引起刮风断裂。主干树皮下明显呈黑褐色坏死,并造成感染面周围树皮开裂,溃疡处一般可看到流胶,严重时造成整株干枯死亡。调查发现,该病发病率通常在 10％～25％,重者可达 40％ 以上,引起桉树溃疡和枯萎死亡。本文通过病原菌分离、致病性测定及传统、分子鉴定明确了该病的病原,以期为病害防治提供理论依据。