Identification and Characterization of Fungal Pathogens Associated with Boxwood Diseases in the Republic of Korea

Boxwood is a representative ornamental shrub that is widely used in landscaping horticulture. After pruning, damaged leaves or stems of boxwoods are unavoidably vulnerable to infection by various plant pathogens. Several boxwood diseases caused by fungi, such as Volutella blight and Macrophoma leaf spot, have been reported worldwide including Republic of Korea. In this study, we isolated and identified fungal pathogens of boxwood diseases that occurred in Korea and characterized their morphological and taxonomic characteristics. Boxwood samples showing blight symptoms were collected in Seoul, Republic of Korea, and the putative fungal pathogens Pseudonectria buxi, P. foliicola, and Neofusicoccum buxi were successfully identified. Investigation of the morphological features of the field isolates, including mycelial growth and conidial morphology, and phylogenetic analysis of multiple DNA barcode loci revealed that there were some morphological and genetic variations among isolates, but all of the analyzed isolates were closely related to the corresponding reference strains. We also found that P. foliicola strains were more virulent than P. buxi, and the N. buxi strains isolated in this study were weak pathogens or saprophytes. The results of our study will contribute to the development of control strategies for boxwood diseases caused by fungi and accelerate research on the complex ecology of boxwood diseases.     

Phylogenetics and evolution of phyllotaxy in the Solomon's seal genus Polygonatum (Asparagaceae: Polygonateae)

Polygonatum is the largest and most complex genus in tribe Polygonateae, comprising approximately 57 species widely distributed in the warm temperate, subtropical and boreal zones of the Northern Hemisphere. However, phylogenetic relationships in the genus remain poorly understood. The objectives of this study were to reconstruct the phylogenetic relationships of the genus using four plastid markers, and to examine the evolution of leaf arrangement in Polygonatum in the phylogenetic context of its closely related taxa. Thirty Polygonatum species were sampled to infer phylogenetic relationships using maximum‐likelihood and Bayesian analyses. The evolution of leaf arrangements was reconstructed using Bayesian, parsimony and likelihood methods. The phylogenetic analyses supported the current generic delimitation of Polygonatum, with Heteropolygonatum recognized as a distinct genus. Three major lineages in Polygonatum were well supported, largely correlated with geographical distribution and the most recent classification at the sectional level. However, our results did not support the currently recognized series, especially the two large series Verticillata and Alternifolia. Bayesian analyses support the alternate‐leaf arrangement as the ancestral state for Polygonatum, but parsimony and maximum‐likelihood analyses suggest an equivocal state for crown Polygonatum. Leaf arrangement was found to be evolutionarily labile. A new nomenclatural combination was made: P olygonatum section S ibirica (L.I.Abramova) Y.Meng, comb. nov. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176 , 435–451.     

Identification of the New Pathogen (Stemphylium lycopersici) Causing Leaf Spot on Pepino (Solanum muricatum)

Pepino (Solanum muricatum var. pepino) plants were found affected by an extensive leaf spot caused by plant pathogenic fungi during a survey in the Cameron highlands, Pahang state, Malaysia. Symptomatic leaf samples were collected from infected pepino plants and cultivated on PDA medium, and the pathogen was isolated and purified; then, consequently, all isolates were identified as Stemphylium lycopersici on the basis of their cultural and morphological characteristics and combined sequences of the internal transcribed spacer (ITS) and glyceraldehyde‐3‐phosphate dehydrogenase (gpd) regions. A pathogenicity assay on detached leaves further confirmed that S. lycopersici causes leaf spot disease. To the best of our knowledge, this is the first report of S. lycopersici causing leaf spot on pepino in Malaysia and worldwide.     

Identification and Characterization of Paraphoma chrysanthemicola Causing Leaf Spot Disease on Atractylodes japonica in China

Atractylodes japonica is a perennial herb in Compositae family, which is used for stomach disorders as a traditional Chinese medicine (Guo et al. 2006 ). In 2013, a leaf spot disease was first observed on plants of A. japonica in a production field of Fushun County, Liaoning Province, China. The disease had a speckled appearance initially. Lesions with grey–white centre and brown margin gradually developed and enlarged. Eventually, infections usually caused yellowing of the leaves and premature defoliation. The causal agent of infection on plants was identified as Paraphoma chrysanthemicola based on morphological and cultural characteristics, pathogenicity tests and phylogenetic analysis. To our knowledge, this is the first report of a leaf spot disease on A. japonica caused by P. chrysanthemicola in China.     

Isolation and Identification of Pathogens Causing Marigold (Tagetes erecta L.) Black Spot in Beijing,China

Black spot leads to great marigold losses worldwide. The disease is characterized by black spots on leaves and stems in its early stages, and the whole plant has black rot at the advanced stage. In this report, 6 of 217 Alternaria strains isolated from lesions of marigold plants in Beijing were randomly selected. The morphological characteristics and a pathogenic tree based on two protein‐coding genes (gpd and alt a 1) indicated that Alternaria tagetica is the causal agent of marigold black spot in Beijing. All six Alternaria strains could successfully re‐infect marigold, but they could not infect carrot or zinnia by either spore spray in a greenhouse or planting experiments in the epidemic area. This is the first report of the A. tagetica pathogen being isolated from marigold in Beijing.     

Taxonomy of five species of cyrtophorids (Protozoa: Ciliophora) including consideration of the phylogeny of two new genera

Cyrtophorids are a specialized group of ciliated protozoa with multitudinous morphotypes. In the present work, the morphology and infraciliature of two new and three rarely known species, including two new genera of cyrtophorid ciliates, Heterohartmannula fangi gen. et sp. nov. , Aporthotrochilia pulex (Deroux, 1976) gen. et comb. nov. , Trochilia alveolata sp. nov. , Trochochilodon flavus Deroux, 1976, and Hypocoma acinetarum Collin, 1907, are described. Heterohartmannula gen. nov. is mainly characterized by a combination of features: two circumoral kineties obliquely arranged, podite not surrounded by somatic kineties, and no distinct gap between left and right ciliary field. Aporthotrochilia gen. nov. is diagnosed mainly by: podite present, oral ciliature reduced to two fragments, several kinety fragments positioned on the right posterior of frontoventral kineties and several terminal fragments. Phylogenetic analyses based on the small subunit rRNA (SSU rRNA) gene sequences support the establishment of two new genera and indicate that Heterohartmannula is most closely related to Hartmannula, and Aporthotrochilia is basal to the Cyrtophoria‐Chonotrichia clade. Trochilia alveolata sp. nov. differs from its congeners mainly by having a conspicuous alveolar layer. In addition, detailed live and infraciliature data of Hypocoma acinetarum and Trochochilodon flavus are supplied. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164 , 1–17.     

Description and identification of two new diseases of guariroba palm (Syagrus oleraceae) in Brazil

Guariroba is a palm species native to central Brazil. Seedlings of guariroba with leaf spots of unknown aetiology were found in Patos de Minas, Minas Gerais, Brazil. The leaf spots were manifest as two different symptom types: the first lesion type consisted of necrotic spots with a rounded to elongate shape, with a light colour and dark edges; the second lesion type had a rounded shape, was dark brown in colour with a light brown edge. The objective of this study was to elucidate the aetiology of both diseases. The likely causal agents were isolated and Koch's postulate fulfilled. Subsequently, the ITS region of rDNA from both micro‐organisms were amplified and sequenced. According to the morphological characteristics and molecular analyses, the fungal species were identified as Pestalotiopsis adusta (causing necrotic spots with a rounded to elongate shape, with a light colour and dark edges), and Alternaria tenuissima (causing lesions with a rounded shape, dark brown in colour with a light brown edge). Identification of the causal organisms of these two diseases will help guide management approaches that might be tested to reduce impact of the disease on Guariroba, including the use of fungicides and cultural approaches.     

Relationships between phoma leaf spot and development of stem canker (Leptosphaeria maculans) on winter oilseed rape (Brassica napus) in southern England

Models were constructed to describe the relationships between incidence of phoma leaf spot at different growth stages in autumn/winter or early spring and incidence of stem canker (basal canker or stem lesions) in summer on winter oilseed rape in southern England. Model 1, describing the phoma leaf spot/basal canker relationship, was y₁=β₀+β₁x₁+β₂(x₂ ‐ x₁) if x₂ > x₁, and y₁=β₀+β₁x₁ if x₂≤x₁, in which y₁ was the incidence (% plants affected) of basal canker at harvest, x₁ was the maximum incidence of phoma leaf spot during the period from sowing to growth stage (G.S.) 1,6‐1,7 (about 100 days after sowing) and x₂ was the maximum incidence of phoma leaf spot between G.S. 1,7 and G.S. 2,0 (start of stem extension). Model 2, describing the phoma leaf spot/stem lesion relationship, was y₂=α₀+α₁x₃+α₂x₄, in which y₂ was the incidence of stem lesions at harvest, x₃ was the incidence of phoma leaf spot at G.S. 3,3–3,5 (flower buds visible) and x₄ was the incidence of phoma leaf spot at G.S. 4,5–5,5 (flower buds opening). Data from field experiments with four winter oilseed rape cultivars at Boxworth or Rothamsted in the 1992/93, 1993/94, 1996/97, 1997/98 or 1998/99 seasons were used to test the models. The values of R² for the regression equations testing model 1 for the phoma leaf spot/basal canker relationship were 0.75, 0.93, 0.91 and 0.89 for cvs Apex, Bristol, Capitol and Envol, respectively. The values of R² for the regression equations testing model 2 for the phoma leaf spot/stem lesion relationship were 0.58, 0.57, 0.54 and 0.71 for cvs Apex, Bristol, Capitol and Envol, respectively. The phoma leaf spot/basal canker relationship (model 1) could also be fitted to the combined data set for all four cultivars (R²= 0.65), whereas the phoma leaf spot/stem lesion relationship (model 2) could not to be fitted to the combined data set for the four cultivars. The relationships between incidence and severity of stem canker were examined and the values of R² for the regressions of severity on incidence were 0.91 for basal canker and 0.89 for stem lesions.     

Characterization and Pathogenicity of Colletotrichum truncatum Causing Stem Anthracnose of Red‐Fleshed Dragon Fruit (Hylocereus polyrhizus) in Malaysia

During August 2010 and January 2011, 10 isolates of Colletotrichum were recovered from stem anthracnose lesions of Hylocereus polyrhizus in the states of Kedah and Penang, Malaysia. Based on the morphological characteristics of colony colour and appearance, and shapes of conidia as well as sequences of internal transcribed spacer regions (ITS), β‐tubulin, actin (ACT) and glyceraldehyde 3‐phosphate dehydrogenase (GAPDH), the fungus was identified as Colletotrichum truncatum. Pathogenicity test showed that C. truncatum isolates were pathogenic to the artificially inoculated H. polyrhizus stem. This is the first report of C. truncatum causing anthracnose on H. polyrhizus stems in Malaysia.     

Identification and Molecular Characterization of a New Geminivirus Betasatellite Infecting Malvastrum coromandelianum in China

The complete nucleotide sequence of a satellite molecule associated with Malvastrum leaf curl Guangdong virus (MLCuGdV) infecting M. coromandelianum plants exhibiting leaf curl symptoms in a suburb of Guangzhou, Guangdong Province of China, is described and analysed. The molecule has typical features of betasatellites, containing a single ORF in the complementary‐sense strand, an A‐rich region, the satellite‐conserved region and a stem–loop structure. Compared with the geminivirus betasatellites in GenBank database, this molecule shows the highest nucleotide sequence identity of 71.9% with Tomato leaf curl Philippine betasatellite isolate Laguna1 (ToLCPB, AB307732). Phylogenetic analysis indicates that it is more related to isolate Laguna 1 and Laguna 2 of ToLCPB. According to the proposed species demarcation threshold of betasatellites (78% nucleotide identity), it is a novel betasatellite species, for which we propose the name Malvastrum leaf curl Guangdong betasatellite (MLCuGdB).     

Molecular Characterization and Distribution of Apple Chlorotic Leaf Spot Virus on Apple in China

Apple chlorotic leaf spot virus (ACLSV) is one of the most economically important latent viruses infecting apple in China. This is the first report of the almost complete nucleotide sequence and the characterization of the genome of a Chinese isolate (ACLSV‐MS, GenBank Accession Number KC847061 ) from apple. Based on the genome nucleotide sequence, ACLSV‐MS showed the highest identity (99.4%) to isolate ACLSV‐B6 (GenBank Accession Number AB326224 ) from apple in Japan and the least identity (69.5%) with isolate TaTao5 (GenBank Accession Number: EU223295 ) from peach in the USA. The occurrence and distribution of ACLSV in China were also recorded. Three hundred and twenty‐seven apple samples (40 different cultivars) collected from 56 sites in 13 provinces of China were tested by RT‐PCR. The virus was detected in all regions surveyed (the provinces of Heilongjiang, Liaoning, Hebei, Beijing, Henan, Shanxi, Shaanxi, Shandong, Gansu, Ningxia, Xinjiang, Sichuan and Yunnan), with an average incidence of 69.7%. The positive samples in Heilongjiang province were highest with an incidence of 100% followed by Henan province with an incidence of 86.7%. The positive samples in Liaoning and Shanxi were the lowest with an incidence of 50%. The occurrence of virus in five common cultivars was determined. The percentage of ACLSV was highest in cv. Gala with an incidence of 33.3%, while lowest in cv. Starking with an incidence of 18.2%. It was also found in younger (≤20 years) apple orchards the occurrence of ACLSV decreased with the increase of tree age, but when trees were more than 20 years old, the occurrence of ACLSV increased. This is the first extensive survey in the last decade in China for monitoring ACLSV, which provides important information for ACLSV control in China.     

A new genus and two new species of Canthocamptidae (Copepoda,Harpacticoida) from caves in northern Italy

Lessinocamptus n. gen. is established toaccomodate three species from northern Italian caves.It can be ascribed to the family Canthocamptidae Sars1906 and is distinguished from the other genera by thecombined characters of: antenna exopod 1-segmented andbearing 3 setae (or 1 seta and 1 spine); mandibularpalp 2-segmented with distal article bearing 5 setae;swimming legs exopods 3-segmented; setal formula ofexopods: P1 0.0.022; P2 0.1.112; P3 0.1.212; P4 0.1.212; P1endopod 2-segmented; P2 endopod 1- or2-segmented; female P3 endopod 1- or 2-segmented, maleP3 endopod 3-segmented and modified as usual inCanthocamptinae; male P5 baseoendopod with 2 spinesand exopod with 3 spiniform setae. Three species aredescribed in detail: L. caoduroi n.sp. from thevadose zone of Lessinian caves, L. insoletus(Chappuis 1928) and L. pivai n.sp. from caveBuso della Rana near Vicenza.     

Identification of Alternaria tenuissima causing brown leaf spot on Paris polyphylla var. chinensis in China

Severe brown leaf spot disease was observed on Paris polyphylla var. chinensis in Sichuan Province, China, in 2017 and 2018. The initial symptoms were many light‐brown small spots with necrotic centres, round or irregular in shape, becoming dark brown, gradually enlarging and eventually coalescing, causing extensive leaf senescence. A fungus was isolated from diseased leaves showing typical symptoms of brown leaf spot. The isolates were cultured on potato sucrose agar, and their morphological characteristics of the causal pathogen were observed under a light microscope. Pathogenicity tests revealed that this fungus was the causal pathogen of the disease. Molecular analyses of the sequences of the ribosomal DNA internal transcribed spacer (ITS) region, translation elongation factor 1‐alpha (TEF) and the RNA polymerase II second largest subunit (RBP2) gene were conducted to confirm the identity of the pathogen. The multi‐gene phylogeny indicated that the causal agent was Alternaria tenuissima. To our knowledge, this is the first report of A. tenuissima causing brown leaf sports on P. polyphylla var. chinensis in China.     

Identification of Colletotrichum Species Causing Bitter Rot of Apple and Pear in Croatia

Symptoms of bitter rot were observed on apple and pear fruits in the field and in storage in Croatia between 2009 and 2011. Fifteen Colletotrichum isolates from apple and two from pear were collected and identified by sequencing of the ITS1 and ITS2 regions of the ribosomal DNA. Phylogenetic analysis revealed that ten isolates from apple and two isolates from pear could be identified as Colletotrichum fioriniae, five isolates from apple were clustered in Colletotrichum clavatum, while one isolate was in the Colletotrichum acutatum A7 group. All isolates caused typical bitter rot symptoms when inoculated on apple and pear fruits.     

中国蓼科萹蓄属植物花粉形态及其分类意义

利用扫描电镜研究了中国萹蓄属(Polygonum) 13种、1变种、1变型植物的花粉形态.结果表明,中国萹蓄属植物的花粉粒为长球形或超长球形,具三孔沟,外壁纹饰可分为4种类型(1)花粉粒表面具细网状纹饰;(2)花粉粒表面具颗粒纹饰;(3)花粉粒表面仅两端具颗粒纹饰,其余部分光滑;(4)花粉粒表面仅沟间区两端肩部光滑,其余部分具颗粒纹饰.根据花粉形态类型,结合该属植物的习性、叶柄基部及花柄上关节的有无、花柱和柱头的数目以及瘦果形态等特征,参照前人的研究处理意见,将中国萹蓄属植物划分为4个组,即小灌木组section Fruticulosae (Boiss.) F.Z.Li et Y.T.Hou,stat.& comb.nov.、萹蓄组section Polygonum、丝茎萹蓄组section Pseudomollia Boiss.和铁马鞭组section Plebeia Tzvel..     

The larval morphology of Arrenurus nobilis Neuman, 1880 and A. robustus Koenike, 1894 (Acari: Parasitengona: Arrenuridae)

The morphology of the larval stages of Arrenurus nobilis and A. robustus is described. Particular attention is given to characters that help to distinguish the two species, and to characters defining the subgenus Arrenurus s. str. The larva of A. robustus resembles the larvae of A. papillator and A. pustulator regarding the shape of dorsal plate, excretory pore plate, coxal plate median margins ratios, and the presence of the IIITa8 seta. The shape of the excretory pore plate in A. nobilis resembles the one in Arrenurus (Megaluracarus) mediorotundatus. The shape of dorsal plate and chelicerae is similar to the ones in the Megaluracarus-species A. globator and A. tubulator. The lack of the IIITa8 seta differs from other species of the subgenus Arrenurus s. str. In the pectinate Lp2 and Mh1 setae A. nobilis resembles representatives of the subgenus Arrenurus s. str. This demonstrates the necessity of verifying the taxonomic status of A. nobilis.     

Identification and Characterization of Pothos latent virus Causing Necrotic Ringspots and Line Patterns on Lisianthus (Eustoma grandiflorum) in Taiwan

Lisianthus (Eustoma grandiflorum) grown in screenhouses in Taiwan showed ringspots and concentric line patterns on leaves. A virus having isometric particles approximately 30–32 nm in diameter was isolated from affected lisianthus. Combined results of biological, cytological, serological, molecular and phylogenetic analyses show that the virus can be identified as Pothos latent virus (PoLV), genus Aureusvirus, family Tombusviridae. Inoculating the virus on non‐infected lisianthus plants reproduced the symptoms previously observed in the field. So, this is the first report of PoLV causing disease in lisianthus and the first report of the virus in Taiwan.     

苹果炭疽叶枯病菌致病力分析及苹果种质抗病性鉴定

苹果炭疽叶枯病是由胶孢炭疽菌(Colletotrichum gloeosporioides)引起的一种真菌病害,现已上升为世界苹果生产中的主要病害之一。了解不同来源的苹果炭疽叶枯病菌致病力差异及明确苹果种质资源对苹果炭疽叶枯病的抗性,对品种选育、品种合理布局以及控制病害的流行具有重要的参考价值。本研究对不同来源的79株病原菌进行了室内致病力测定,获知该菌致病力差异明显,其中强致病力菌株所占比例大。同时,本研究也对327份苹果种质资源进行了室内抗病性鉴定,其中高抗资源160份,中抗资源6份,中感资源22份,高感资源139份。表明我国现保存的苹果种质资源中存在丰富抗病种质。进一步按苹果分类系统分析发现,抗病资源在当前栽培的主要品种群中均有分布,特别是红玉品种群、富士品种群抗病资源最为丰富。