Inhibitory effects of essential oil components extracted from celery (Apium graveolens) on the growth of Septoria apiicola, causal agent of leaf spot disease

Leaf spot disease of celery ( Apium graveolens ), caused by the fungus Septoria apiicola , is a major disease for which there is only limited host resistance. Some factors affecting the response of the fungus to essential oil components of host tissue were investigated using a susceptible blanching celery (var. Celebrity) and a more resistant green celery (var. Cutting) as hosts. After infection the time taken for pycnidia to appear on the surface was 16 days in Cutting and 13 days in Celebrity, but only limited leaf necrosis occurred in Cutting whereas the whole leaf rapidly became necrotic in Celebrity. Although the composition of the essential oil (limonene, β-pinene, terpinene, caryophyllene, sesquiterpenes and phthallides) was comparable in both varieties, Cutting contained over four times the total concentration of oil found in Celebrity. Extracts of the essential oil from Cutting were found to be more inhibitory to the growth of Septoria in culture. Of the terpenes tested (limonene, β-pinene, and caryophyllene), β-pinene was most inhibitory to the growth of hyphae on solid and liquid media. The role of the essential oil as part of a constitutive defense mechanism in host resistance to Septoria is discussed.     

Cylindrocladium brown leaf spot on Howea belmoreana caused by Calonectria ilicicola (anamorph: Cylindrocladium parasiticum) in Japan

Brown leaf spot disease caused by Cylindrocladium was found on Howea belmoreana on Hachijojima Island, Tokyo, Japan, in December 2001. Typical symptoms were incited after artificial inoculation. A culture of white mycelia, isolated from leaf spot symptoms, produced reddish perithecia of a nectriaceous fungus. Based on morphological and molecular analyses, this fungus was identified as Calonectria ilicicola (anamorph: Cylindrocladium parasiticum). Pathogenicity of this fungus on five plants cultivated on Hachijojima Island was confirmed by artificial inoculation. This report is the first on Cylindrocladium brown leaf spot of H. belmoreana caused by C. ilicicola (anamorph: Cy. parasiticum).     

Genetic Structure of Mycosphaerella populorum (Anamorph Septoria musiva) Populations in North-Central and Northeastern North America

ABSTRACT In order to characterize the genetic variation of the poplar pathogen Mycosphaerella populorum (anamorph Septoria musiva), we have studied seven North American populations using the polymerase chain reaction random amplified polymorphic DNA (RAPD) technique. The fungal populations were sampled in 2001 and 2002 by obtaining 352 isolates from cankers and leaf spots in hybrid poplar plantations and adjacent eastern cottonwood (Populus deltoides). A total of 21 polymorphic RAPD markers were obtained with the six RAPD primers used. A fine-level scale analysis of the genetic structure within the populations revealed that subpopulations sampled on P. deltoides and on hybrid trees were not significantly differentiated. In contrast, analyses performed on the entire data set showed high levels of haplotypic diversity and moderate to high genetic differentiation, with 20% of the expected genetic diversity found at the interpopulation level. Moreover, a high and significant correlation between genetic and geographic distances among populations was found, suggesting isolation by distance of the sampled populations. Although the occurrence of the sexual stage of this fungus remained unclear in field populations, five of the six populations were at gametic equilibrium for RAPD loci, suggesting the occurrence of recombination episodes in Septoria musiva populations. Overall, S. musiva appears to consist of differentiated subpopulations, with both asexual and sexual recombination contributing to the local level of genetic structure.     

First record of onion leaf blotch (Mycosphaerella allii-cepae) in Argentina

In 1990, onion leaf blotch was observed for the first time in the Lower Rio Colorado Valley (Province of Buenos Aires), the most important onion-producing region of Argentina. Since then, the disease has been found in other production areas of this country. Infections on shallot have also been recorded. The causal agent, Mycosphaerella allii-cepae , was identified on the basis of symptoms, fungus morphology (as its anamorph Cladosporium allium-cepae ) and successful artificial inoculations on onion plants. Although the disease has been present in all seasons since 1990, epidemics of any importance have not been observed in the main crop of dry onions. This is the first record of onion leaf blotch for Argentina.     

Disease Note: Identification of Curvularia oryzae as cause of leaf spot disease on oil palm seedlings in nurseries of Thailand

The causal agent of leaf spot disease of oil palm (Elaeis guineensis) seedling nurseries in Thailand was identified as Curvularia oryzae. The fungus was isolated from leaves with disease symptoms, characterized by morphological properties, and pathogenicity tested. The identity of the phytopathogenic fungus was confirmed through polymerase chain reaction analysis using internal transcribed spacer (ITS) primers, which amplified about a 1 kb product. Sequencing this DNA product confirmed this pathogen was C. oryzae. Furthermore, the pathogenicity test showed that C. oryzae could infect oil palm seedlings.     

Characterization and pathogenicity of Septoria gaurina associated with leaf blotch disease on Gaura parviflora in China

Gaura parviflora, a flowering plant of the family Onagraceae, is regarded as an invasive plant in China, but is subject to leaf blotch disease. Based on morphological features, we identified the causal agent to be Septoria gaurina. However, the phylogenetic position of S. gaurina remains unknown. Multilocus phylogenetic analysis using the internal transcribed spacer (ITS), β-tubulin (Tub2), actin (Act), calmodulin (Cal), translation elongation factor 1-α (EF), and 28S nrDNA (LSU) nucleotide sequences indicated that the isolates belonged to the Septoria genus. Pathogenicity tests demonstrated that the isolates produced symptoms on the leaves of G. parviflora. These symptoms included small, water-soaked, circular lesions formed on leaves inoculated with conidial droplets, and bigger, water-soaked lesions on leaves inoculated by conidial spraying. To our knowledge, this is the first report of S. gaurina as the causative agent of leaf blotch disease of G. parviflora in China.     

Molecular diagnostics for the sigatoka disease complex of banana

ABSTRACT The Sigatoka disease complex of banana involves three related ascomycetous fungi, Mycosphaerella fijiensis, M. musicola, and M. eumusae. The exact distribution of these three species and their disease epidemiology remain unclear, because their symptoms and life cycles are rather similar. Disease diagnosis in the Mycosphaerella complex of banana is based on the presence of host symptoms and fungal fruiting structures, which hamper preventive management strategies. In the present study, we have developed rapid and robust species-specific molecular-based diagnostic tools for detection and quantification of M. fijiensis, M. musicola, and M. eumusae. Conventional species-specific polymerase chain reaction (PCR) primers were developed based on the actin gene that detected DNA at as little as 100, 1, and 10 pg/mul from M. fijiensis, M. musicola, and M. eumusae, respectively. Furthermore, TaqMan real-time quantitative PCR assays were developed based on the beta-tubulin gene and detected quantities of DNA as low as 1 pg/mul for each Mycosphaerella sp. from pure cultures and DNA at 1.6 pg/mul per milligram of dry leaf tissue for M. fijiensis that was validated using naturally infected banana leaves.     

Research on Septoria Leaf Blotch: Recent Advances1

A review of the recent advances in research on Septoria leaf blotch is given with particular emphasis on epidemiology and control strategies. The effect of straw infected with pycnidia of Septoria tritici or perithecia of its perfect stage Mycosphaerella graminicola , infected seeds and alternative hosts on the initiation of epidemics is discussed. Reference is made to the effect of plant stature on the vertical progress of the disease. The role of fungicide protection in the control of the disease is discussed. Resistance and tolerance to Septoria leaf blotch is dealt with in more detail, with reference to the author's own research, and with implications on breeding strategies.     

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.     

Phenotypic and genetic variability of fungal isolates associated with the Septoria leaf spot disease of lettuce (Lactuca sativa) in Brazil

Journal of Plant Diseases and Protection - Septoria leaf spot is a severe disease of lettuce in tropical and subtropical regions, inducing yield/quality losses and increasing production costs....     

First report of leaf spot caused by <Emphasis Type="Italic">Alternaria alternata</Emphasis> on <Emphasis Type="Italic">Drimia maritima</Emphasis>

Drimia maritima (squill) is a historically important medicinal plant. During the spring of 2016, small, yellow leaf spots, which became brown and finally necrotic, were observed on squill plants in Kohgiluyeh and Boyer-Ahmad Provinces in Iran. A fungus was consistently isolated from infected leaves and identified as Alternaria alternata based on morphological and phylogenetic analyses. Pathogenicity tests confirmed A. alternata to be the causal agent of the newly observed leaf spot disease. This is the first report of leaf spot on D. maritima caused by A. alternata in the world.     

Modeling of Relationships Between Weather and Septoria tritici Epidemics on Winter Wheat: A Critical Approach

ABSTRACT Two models for predicting Septoria tritici on winter wheat (cv. Riband) were developed using a program based on an iterative search of correlations between disease severity and weather. Data from four consecutive cropping seasons (1993/94 until 1996/97) at nine sites throughout England were used. A qualitative model predicted the presence or absence of Septoria tritici (at a 5% severity threshold within the top three leaf layers) using winter temperature (January/February) and wind speed to about the first node detectable growth stage. For sites above the disease threshold, a quantitative model predicted severity of Septoria tritici using rainfall during stem elongation. A test statistic was derived to test the validity of the iterative search used to obtain both models. This statistic was used in combination with bootstrap analyses in which the search program was rerun using weather data from previous years, therefore uncorrelated with the disease data, to investigate how likely correlations such as the ones found in our models would have been in the absence of genuine relationships.     

First report of celery stunt anthracnose caused by <Emphasis Type="Italic">Colletotrichum simmondsii</Emphasis> in Japan

A new disease was found in Japan, on celery (Apium graveolens var. dulce) having severe chlorotic leaf spot, stunt, and dwarf with leaf curl. A spore suspension from the fungus isolated from affected plants induced identical symptoms 14 days after plants were sprayed. Identification and molecular characterization showed that the causal agent is Colletotrichum simmondsii. This report is the first of stunt anthracnose on celery caused by C. simmondsii. We propose the name “stunt anthracnose” for the new disease. Colletotrichum acutatum sensu lato, as reference pathogen of celery anthracnose, should be changed to C. fioriniae based on morphological and molecular characteristics.     

Effects of Water Stress on Colonization of Poplar Stems and Excised Leaf Disks by Septoria musiva

ABSTRACT Septoria musiva causes leaf spot and canker diseases of trees in the genus Populus, and is one of the most damaging fungal pathogens of hybrid poplar in eastern North America. The effect of host water stress on Septoria canker development was studied in two separate greenhouse experiments. Hybrid poplar clones NM6, NC11396, and NE308 were stressed by withholding water until predawn water potential fell below -1.0 MPa. Stems were treated by removing a leaf and applying agar plugs that were either colonized by S. musiva (inoculated) or sterile (control) to the wound. Cankers on inoculated water-stressed trees were significantly larger than those on nonstressed trees. A leaf disk assay also was conducted three times with the NM6 and NE308 trees. We cut two disks from each of 120 stressed and 120 well-watered trees, placing them on water agar in 24-well tissue culture plates. A conidial suspension was applied to one disk in each pair and sterile water to the other. Inoculated disks from water-stressed trees developed less necrosis than those from well-watered trees. These results demonstrate that environmental influences on host condition must be considered in evaluating resistance of clones proposed for widespread culture of hybrid poplar.     

A method of inoculating barley seedlings with Ramularia collo‐cygni

A method of inoculating seedlings with the fungus Ramularia collo-cygni, the causal agent of ramularia leaf spot (RLS), an increasingly important problem in barley in Europe and elsewhere, is described. Symptoms of RLS similar to those found in the field were reproduced on seedlings and the fungus was re-isolated from them, fulfilling the third and fourth of Koch's postulates. The method was similar to one used for the related fungus, Mycosphaerella graminicola (anamorph Septoria tritici), a pathogen of wheat. Briefly, plants were sprayed with a suspension of R. collo-cygni mycelium fragments, incubated at 15°C, first in darkness for 48 h then in a 16-h-light/8-h-dark cycle. Disease levels reached saturation when plants were sprayed to runoff with a suspension of 480 cm² of mycelium, scraped from the entire surface of 7·5 Petri dishes (9 cm diameter) and sieved, in 50 mL water. Growth of seedlings in high light intensity (900 µmol m⁻² s⁻¹, 16-h daylength) before inoculation increased disease symptoms, but reduced disease when applied after inoculation. In contrast to M. graminicola, near-ultraviolet light after inoculation reduced symptom development. It is proposed that for the full development of RLS, plants should be grown in a stressful environment before inoculation. Nine barley lines were assessed for their resistance to RLS as seedlings and a subset were tested in field trials with natural infection by R. collo-cygni. There was cultivar-by-isolate interaction in the amount of RLS symptoms on seedlings. RLS levels on adult plants in field plots were correlated with RLS scores on seedlings formed by one isolate but not the other.     

应用RAPD标记技术鉴定香蕉褐缘灰斑病菌

采用Mycosphaerella fijiiensis和M.musicola的RAPD标记技术鉴定海南香蕉褐缘灰斑病菌,结果表明,分离自海南儋州、乐东、文昌、东方、澄迈、临高、琼海、昌江、琼山、三亚香蕉上的10个菌株均为M.fijiensis,引起香蕉黑叶条斑病。     

Black leaf spot of Japanese persimmon (<Emphasis Type="Italic">Diospyros kaki</Emphasis>), a new disease caused by <Emphasis Type="Italic">Adisciso kaki</Emphasis> sp. nov.

A new defoliating disease with severe leaf spot of Japanese persimmon (Diospyros kaki) was found in Shimane Prefecture. A fungus, isolated from the ascospores that had formed on the leaf spot, was demonstrated to reproduce the symptoms on leaves and described as a new species, Adisciso kaki, based on morphological and molecular phylogenetic analyses. This fungus is characterized by small ascomata without clypeus, obclavate to broadly cylindrical asci with an amyloid apical apparatus, and hyaline ascospores with a submedian septum. We coined the name black leaf spot (Kurohoshi-rakuyo-byo, in Japanese) for the new disease.     

新疆加工型辣椒细菌性斑点病的发生和病原鉴定

 新疆加工型辣椒主要产区巴音郭楞蒙古自治州发生了一种严重危害辣椒的细菌性病害。从发病辣椒叶片中分离细菌,通过烟草过敏性反应、马铃薯软腐试验和接种辣椒等致病性测定,确定了13个致病菌株,各菌株之间致病力无明显差异。通过菌体形态、培养性状观察、生理生化反应、寄主范围测定,结合16S rDNA和rpoD基因扩增、序列测定和系统发育分析,将病原菌鉴定为丁香假单胞菌丁香致病变种(Pseudomonas syringae pv. syringae)。病原菌人工接种还能侵染番茄、茄子、马铃薯及黄瓜、四季豆、白菜、萝卜、芹菜等植物。P. syringae pv. syringae引起加工型辣椒细菌性斑点病在国内属首次报道。     

Factors affecting development of Septoria tritici in winter wheat and its effect on yield

Severe infection with Septoria tritici occurred in four of five experiments designed to create a series of different disease epidemics. These experiments successfully identified periods suitable for infection. They also indicated the effect of sprays, timed before and after these periods, on disease development and yield.
Analysis of disease progress and weather records suggested that critical conditions for initial development of S. tritici occurred during early May at four sites. Heavy rain giving at least 10 mm on 1 day or a total of 10 mm or more on 2 or 3 successive days occurred at all four sites prior to the appearance of symptoms on a particular leaf layer, though less heavy rain may suffice to splash inoculum onto upper leaves in shorter, immature canopies where stem elongation is incomplete. At the fifth site, infection occurred later in May and disease failed to develop to a significant degree. At all sites, the length of the incubation period on any of the top three leaves was found to be between 396 and 496 degree days.
Control of winter epidemics of S. tritici had little effect on yield, whereas spray sequences commencing later than growth stage (GS) 31 but immediately prior to the critical periods provided the best disease control and yield benefit. Regression models incorporating, as independent variables, area under the S. tritici disease progress curve for any of the top three leaves from their emergence (GS 32-37) satisfactorily explained yield loss at the four sites where disease was severe. Consideration of leaf 2 or leaf 3 alone accounted for more than 82% of the variance at each site and a yield loss from infection of leaf 2 related to thermal time is suggested as 0.00265% per C per day from the appearance of symptoms.     

Klebsiella pneumoniae subsp. pneumoniae 引起云南省香蕉细菌性叶斑病

 为明确一种新的香蕉（Musa sapientum）细菌性叶斑病病原菌,以云南省新平县香蕉园区发现的一种新病害为供试材料,通过分离培养、形态观察、致病性测定、生理生化试验和gyrB,16S rDNA 和 rpoB基因片段分析,对病原菌进行了鉴定。该病由克雷伯氏肺炎球菌肺炎亚种（Klebsiella pneumoniae subsp. pneumoniae）引起,病菌可侵染香蕉叶片、假茎和果实。茎干被病原菌侵入3 d后即可出现黑色小斑点,在接种部位附近呈上下方向蔓延趋势,7 d后茎干上出现大面积棕色坏死,内部组织褐变。本文在世界上首次报道克雷伯氏肺炎球菌肺炎亚种可侵染香蕉植株,引起香蕉细菌性叶斑病。