Overexpression of a Gene Encoding a Catabolite Repression Element in Alternaria citri Causes Severe Symptoms of Black Rot in Citrus Fruit

ABSTRACT A gene (AcCreA) encoding a catabolite repression element (CreA) with (two zinc fingers of the Cys(2)His(2) type was isolated from the postharvest fungal pathogen Alternaria citri. The AcCreA overexpression mutant AcOEC2 of A. citri showed normal growth on pectin medium and on segments of peel or the juice sac area from citrus fruit. Production of endopolygalacturonase, an essential virulence factor of this pathogen, was similar in AcOEC2 and the wild type in pectin-containing media. However, addition of glucose to the medium showed that carbon catabolite repression of endopolygalacturonase gene (Acpg1) expression, as well as endopolygalacturonase production, was lost in AcOEC2. The wild-type strain of A. citri causes rot mainly in the central axis of citrus fruit without development of rotting in the juice sac area; however, AcOEC2 caused severe black rot symptoms in both the central axis and juice sac areas. These results indicate that AcCreA-mediated catabolite repression controls the virulence or infection of this pathogen, and that the wild-type A. citri does not cause symptoms in the juice sac area due to carbon catabolite repression by sugars in the juice of the juice sac area.     

Citrus Black Rot is Caused by Phylogenetically Distinct Lineages of Alternaria alternata

ABSTRACT Phylogenetic analysis revealed that isolates of Alternaria alternata causing black rot of citrus were associated with six well-supported evolutionary lineages. Isolates recovered from brown spot lesions on Minneola tangelo, leaf spot lesions on rough lemon, and healthy citrus tissue and noncitrus hosts were related closely to isolates from black-rotted fruit. Phylogenies estimated independently from DNA sequence data from an endopolygalacturonase gene (endoPG) and two anonymous regions of the genome (OPA1-3 and OPA2-1) had similar topologies, and phylogenetic analysis was performed on the combined data set. In the combined phylogeny, isolates from diverse ecological niches on citrus and noncitrus hosts were distributed in eight clades. Isolates from all lineages, regardless of ecological or host association, caused black rot in fruit inoculation assays, demonstrating that small-spored Alternaria isolates associated with different ecological niches on citrus and other plant hosts are potential black rot pathogens. These data also indicated that the fungi associated with black-rotted fruit do not form a natural evolutionary group distinct from other Alternaria pathogens and saprophytes associated with citrus. The use of the name A. citri to describe fungi associated with citrus black rot is not justified and it is proposed that citrus black rot fungi be referred to as A. alternata.     

Stone Fruit Tree Apoplexy Caused by Phytophthora Collar Rot

Stone fruit tree apoplexy in Greece is often caused by Phytophthora spp. Two types of Phytophthora apoplexy of stone fruit tree can be distinguished, one occurring during the hot summer period and usually caused by P. cactorum (Leb. et Cohn) Schröt. or P. citrophthora (R. et E. Smith) Leonian, and a second occurring in winter or early spring, and caused by P. syringae (Kleb.) Kleb. or sometimes P. megasperma Drechsler. Inoculation tests showed P. syringae to be highly pathogenic to apricot and almond, whereas certain clonal rootstock material showed resistance. Measures of control against Phytophthora attacks are given.     

Threshold Conditions That Lead Latent Infection to Prune Fruit Rot Caused by Monilinia fructicola

ABSTRACT Inoculations were performed six to eight times in each of 10 prune orchards located in nine counties of California. In each inoculation, branches that bore 40 to 60 blossoms or 30 to 40 fruit were inoculated with conidial suspensions of Monilinia fructicola. Three inoculum concentrations and 14 to 16 h of humidity were used for each inoculation. All inoculated fruit were maintained on trees and harvested separately 2 weeks before commercial harvest. The incidence of latent infection (ILI) and percentage of branches with fruit rot (PBFR) were determined for each inoculation in each orchard. As the ILI increased, the PBFR also increased linearly. Five conditions that lead latent infection to fruit rot include (i) latent infection level; (ii) fruit developmental stage; (iii) inoculum concentration; (iv) total hours of relative humidity greater than 90% (hRH); and (v) total hours of dew period (hDEW) from mid-July to mid-August. Three levels of PBFR, 1, 5, and 10% were assigned, and threshold conditions that lead to these levels were determined based on the experimental results. The relative probabilities that lead latent infection to fruit rot (r_PBFR) at different fruit developmental stages were calculated. A preliminary decision support model to guide fungicide application was developed based on the above results. One of the four recommendations, safe, wait, check historical weather as a reference, and apply a fungicide immediately, could be provided based on the level of latent infection and the decision process developed through this study.     

Etiology and Population Genetics of Colletotrichum spp. Causing Crown and Fruit Rot of Strawberry

ABSTRACT Isolates of Colletotrichum spp. from diseased strawberry fruit and crowns were evaluated to determine their genetic diversity and the etiology of the diseases. Isolates were identified to species using polymerase chain reaction primers for a ribosomal internal transcribed spacer region and their pathogenicity was evaluated in bioassays. Isolates were scored for variation at 40 putative genetic loci with random amplified polymorphic DNA and microsatellite markers. Only C. acutatum was recovered from diseased fruit. Nearly all isolates from crowns were C. gloeosporioides. In crown bioassays, only isolates of C. gloeosporioides from strawberry caused collapse and death of plants. A dendrogram generated from the genetic analysis identified several primary lineages. One lineage included isolates of C. acutatum from fruit and was characterized by low diversity. Another lineage included isolates of C. gloeosporioides from crowns and was highly polymorphic. The isolates from strawberry formed distinctive clusters separate from citrus isolates. Evaluation of linkage disequilibrium among polymorphic loci in isolates of C. gloeosporioides from crowns revealed a low level of disequilibrium as would be expected in sexually recombining populations. These results suggest that epidemics of crown rot are caused by Glomerella cingulata (anamorph C. gloeosporioides) and that epidemics of fruit rot are caused by C. acutatum.     

中国发现甘薯长喙壳菌引起枣子果实腐烂

 2013年8月,在中国昆明发现一种枣子果实的腐烂病。通过致病性测试,证明从发病组织分离出的病菌能引起枣子果实腐烂。经过形态学和分子鉴定,确定该病原菌为甘薯长喙壳菌。这是在中国首次报道由甘薯长喙壳菌引起的枣子果实腐烂病。     

Sensitivity of Red Delicious Apple Fruit at Various Phenologic Stages to Infection by Alternaria alternata and Moldy-core Control

The fungus Alternaria alternata, is considered to be the predominant fungus involved in moldy-core of Red Delicious strains of apple. In this paper, we report on the sensitivity of various phenologic stages to infection by A. alternata, and on the efficacy of various fungicides in controlling moldy-core disease in apple orchards. Artificial inoculations conducted in the orchard during 1999 and 2000 seasons revealed that the beginning of bloom (10–30%) and full bloom were the most susceptible developmental stages for infection. Natural infection with A. alternata in fruits was relatively high, reaching 44% and 46% of the fruits on control non-treated trees in 1999 and 2000. Four foliar applications of polyoxin B, difenoconazole and azoxystrobin, starting from the beginning of bloom until fruit set, reduced the number of infected fruits by 54–70%, 61–70% and 50–65%, respectively, compared with non-treated trees. Four or eight foliar applications of each fungicide provided similar levels of control. There were no significant differences between two, four or six foliar applications of difenoconazole, neither between two or four applications of polyoxin B. Adding CaCl₂, as a tank mixture with difenoconazole at full rate, did not improve efficacy. Alternaria was recovered from the inner part of the core region of 71–88% of the fruits of the non-treated control, but was recovered less frequently from the outside part of the core region. Fruits of difenoconazole and polyoxin B treated trees were less colonized with A. alternata at both the inner and outside parts of the core region, as compared with controls. Results indicate that a control programme based on spray applications of difenoconazole or polyoxin B, during bloom period, can effectively reduce Alternaria on Red Delicious.     

中国玉米茎基腐病和穗腐病研究进展

 本文综述了我国在玉米茎基腐病和穗腐病病原学、致病性和寄主抗性机制与综合治理的研究进展。深入探讨了以往我国学者关于引起茎基腐病病原学的不同观点。根据2种病害的病原镰孢菌在可溶性蛋白质、血清学、同功酶和DNA等不同水平的多态性分析,镰孢菌在寄主根和茎组织内侵染过程示踪和孢子捕捉试验结果,重点讨论了2种镰孢菌病害在病原学侵染规律方面的相互关系,概述了2种病害寄主抗性生理生化机理、抗性遗传和以生物防治为核心的综合治理措施研究进展。     

Resistance of Gibberellin-Treated Persimmon Fruit to Alternaria alternata Arises from the Reduced Ability of the Fungus to Produce Endo-1,4-beta-Glucanase

ABSTRACT Black-spot symptoms, caused by Alternaria alternata, developed in persimmon fruits during prolonged storage at -1 degrees C. A preharvest treatment with gibberellic acid (GA(3)) extended the storage life of the fruit by delaying both black-spot development and fruit softening. Conversely, treatment of persimmon fruits with paclobutrazol (PBZ), an inhibitor of gibberellin (GA) synthesis, enhanced black-spot development and fruit softening during storage. Production of endo-1,4-beta-glucanase (EC 3.2.1.4, EG) by A. alternata in culture and in the presence of cell walls from PBZ-treated fruits as the carbon source, was enhanced by 150% over production in the presence of cell walls from control fruits, whereas endoglucanase (EG) production in the presence of cell walls from GA(3)-treated fruits was reduced by 49% relative to controls. To determine the importance of EG in symptom development, A. alternata EG was purified from a culture-inducing medium. It had a molecular mass of 41 kDa, its optimal pH and temperature for activity were 5.5 and 47 degrees C, respectively, and the pI was 4.3. Its K(m) and V(max) were 0.43 mg ml(-1) and 18 mumol reducing groups minute per milligrams of protein, respectively. The internal sequence of a 21-mer amino acid peptide from the purified EG showed 62% similarity and 38% identity to the EG-1 of Trichoderma reesei and of T. longibrachiatum. Purified EG induced black-spot symptoms on the fruit, similar to those caused by A. alternata, whereas boiled enzyme caused only pricking signs. Our results suggest that the black-spot symptoms caused by A. alternata, in persimmon, are related to the ability of the fungus to produce EG in developing lesions.     

Contrasting species boundaries between sections Alternaria and Porri of the genus Alternaria

The fungal genus Alternaria comprises a large number of asexual taxa with diverse ecological, morphological and biological modes ranging from saprophytes to plant pathogens. Understanding the speciation processes affecting asexual fungi is important for estimating biological diversity, which in turn affects plant disease management and quarantine enforcement. This study included 106 isolates of Alternaria representing five phylogenetically defined clades in two sister sub‐generic groups: section Porri (A. dauci, A. solani and A. limicola) and section Alternaria (A. alternata/tenuissima and A. arborescens). Species in section Porri are host‐specific while species in section Alternaria have wider host ranges. For each isolate, DNA sequences of three genes (Alt a1, ATPase, Calmodulin) were used to estimate phylogenies at the population and species levels. Three multilocus haplotypes were distinguished among A. dauci isolates and only one haplotype among A. solani and A. limicola isolates, revealing low or no differentiation within each taxon and strong clonal structure for taxa in this section. In contrast, 37 multilocus haplotypes were found among A. alternata/tenuissima isolates and 21 multilocus haplotypes among A. arborescens isolates, revealing much higher genotypic diversity and multiple clonal lineages within taxa, which is not typical of asexual reproducing lineages. A species tree was inferred using a Yule Speciation model and a strict molecular clock assumption. Species boundaries were well defined within section Porri. However, species boundaries within section Alternaria were only partially resolved with no well‐defined species boundaries, possibly due to incomplete lineage sorting. Significant association with host specificity seems a driving force for speciation.     

Laboratory Evaluation of Citrus Cultivars Susceptibility and Influence of Fruit Size on Fortune Mandarin to Infection by Alternaria alternata pv. citri

Young leaves of 62 citrus cultivars were inoculated with conidia of three Spanish isolates of Alternaria alternata pv. citri, the causal agent of brown spot of citrus. Hybrids with Dancy mandarin, King mandarin or their derivates as a parent, grapefruit cultivars and the mandarin cultivars Guillermina, Emperor, Clemenpons and Esbal were highly susceptible to the pathogen. Satsuma cultivar Clausellina and orange cultivars, with the exception of Sanguinelli, were slightly susceptible. Lemon and lime cultivars were not susceptible, with the exception of Mexican lime (Citrus aurantifolia), which was slightly susceptible. Although this study shows a range of potential hosts for this pathogen, to date the only affected cultivars in Spain are Fortune and Nova mandarins, and Minneola tangelo. The susceptibility of Fortune fruits decreased as diameter increased, being susceptible through the whole season. This was confirmed with field observations in autumn where fruit infections have been detected when the diameter reaches 6–7 cm.     

Fusarium Redolens f.sp asparagi, Causal Agent of Asparagus Root Rot, Crown Rot and Spear Rot

Two Fusarium species, F. oxysporum f.sp. asparagi and F. proliferatum, are known to be involved in the root and crown rot complex of asparagus. We have investigated reports on the involvement of F. redolens, a third species, which until recently was considered conspecific with F. oxysporum because of morphological similarities. RFLP analysis of the rDNA internal transcribed spacer region and AFLP fingerprinting identified eight strains from asparagus unambiguously as F. redolens. Four of these were tested and found to be pathogenic to asparagus either in this study (two strains) or in a previous one in which they were classified as F. oxysporum (three strains). Disease symptoms and disease development were the same as with F. oxysporum f.sp. asparagi and F. proliferatum. Present data and literature reports identify F. redolens as a host-specific pathogen involved in root, crown and spear rot of asparagus. The pathogen is formally classified as F. redolens Wollenw. f.sp. asparagi Baayen.     

向日葵链格孢的分类研究

引起油葵黑斑病的病原菌在我国一直采用向日葵链格孢Alternaria helianthi这个学名。然而国际上已更名为向日葵拟链格孢Alternariaster helianthi。2017年从陕西省华阴市采集到油葵黑斑病样品。通过单孢分离获得6株病原菌菌株,致病性检测后选取2个代表菌株进行形态学研究,并基于ITS和LSU基因进行系统发育分析。结果表明,2个病原菌菌株为小球腔菌科Leptosphaeriaceae拟链格孢属Alternariaster向日葵拟链格孢Alternariaster helianthi。     

Alternaria Leaf Spot on Three Species of Pelargonium Caused by Alternaria alternata in Japan

A new disease of pelargonium (Pelargonium domesticum Bailey), ivy geranium (P. peltatum (L.) L'Hér. ex Ait.) and scented geranium (P. graveolens L'Hér.), primarily causing brown spots on leaves, was found in Kawasaki-shi in Kanagawa Prefecture and Tachikawa-shi in Tokyo. An Alternaria sp. was consistently isolated from these diseased leaves, and the isolates were pathogenic to their host leaves. Based on morphological characteristics, the causal fungus in all three cases was identified as Alternaria alternata (Fr.) Keissler. Because Alternaria leaf spot of geranium by A. alternata has already been reported, the pathogenicity of isolates from four groups of genus Pelargonium was investigated. The isolates from scented geranium were pathogenic only to their original host, but the isolates from pelargonium, ivy geranium and geranium were pathogenic to all groups of pelargonium. This is the first report of this disease on pelargonium, ivy geranium and scented geranium caused by A. alternata in Japan. We propose the names for these diseases as Alternaria leaf spot of pelargonium (kappan-byo), Alternaria leaf spot of ivy geranium (kappan-byo) and Alternaria leaf spot of scented geranium (kappan-byo). Received 11 December 2000/ Accepted in revised form 19 July 2001     

Effect of nutrients and plant extracts on Alternaria blight of tomato caused by Alternaria Alternata

Journal of Plant Diseases and Protection - Studies with nutrients and phyto-extracts were performed in vitro and in vivo to assess their potential in controlling Alternaria blight of tomato...