scholarly journals First Report of Wood Canker Caused by Botryosphaeria dothidea, Diplodia seriata, Neofusicoccum parvum, and Lasiodiplodia theobromae on Grapevine in Turkey

Plant Disease ◽  
2014 ◽  
Vol 98 (4) ◽  
pp. 568-568 ◽  
Author(s):  
D. S. Akgul ◽  
N. G. Savas ◽  
A. Eskalen
Keyword(s):  
Morphological Characteristics ◽  
Botryosphaeria Dothidea ◽  
Outer Bark ◽  
Western Turkey ◽  
Diplodia Seriata ◽  
Neofusicoccum Parvum ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Multiple Species ◽  
Botryosphaeriaceae Species

The Aegean region (western Turkey) is the center of table, raisin, and wine grape cultivation. During the 2012 growing season, wood canker symptoms were observed in vineyards in Manisa city. Symptoms adjacent to pruning wounds, including shoot dieback and wedge-shaped wood discolorations observed in cross section, were among the most prevalent symptoms of the vines. To identify the causal agents, symptomatic woody tissues were surface disinfested with 95% ethanol and flame-sterilized and the discolored outer bark was cut away. The internal tissues (0.5 cm2) were excised from cankers of vines and plated onto potato dextrose agar amended with tetracycline (0.01%) (PDA-tet). The most frequently isolated fungi, based on general growth pattern, speed of growth, and colony color, resembled species in the Botryosphaeriaceae family. According to morphological characteristics, four different groups have been identified based on visual discrimination. After DNA extraction, ribosomal DNA fragments (ITS1-5.8S-ITS2) (2) amplified with ITS4 and ITS5 primers were sequenced and sequences were compared with those deposited in NCBI GenBank database. Four different Botryosphaeriaceae isolates were identified, including Botryosphaeria dothidea (MBAi25AG), Diplodia seriata (MBAi23AG), Lasiodiplodia theobromae (MBAi28AG), and Neofusicoccum parvum (MBAi27AG) (Accession Nos. KF182329, KF182328, KF182331, and KF182330, respectively) with species nomenclature based on Crous et al. (1). Pathogenicity tests were conducted under greenhouse conditions (24°C, 16/8-h day/night, 70% RH) on 1-year-old own rooted grapevine (Vitis vinifera) cv. Sultana Seedless seedlings using one isolate from each of the Botryosphaeriaceae species specified above. Stems of grapevine seedlings were wounded by removing bark with 4-mm cork borer and fresh mycelial plugs were inoculated into the holes and covered with Parafilm. Sterile PDA plugs were placed into the wounds of control seedlings. Five vines were inoculated per isolate. The experiment was repeated twice. After 4 months of incubation, grapevine seedlings were examined for the extent of vascular discoloration and recovery of fungal isolates. Mean lesion lengths on wood tissues were 85.3, 17.2, 13.9, and 13.1 mm for N. parvum, B. dothidea, L. theobromae, and D. seriata, and 6.3 mm for control. Each fungal isolate was successfully re-isolated from inoculated seedlings to fulfill Koch's postulates. To our knowledge, this is the first report of multiple species in the Botryosphaeriaceae causing wood canker and dieback on grapevine in Turkey. These results are significant because Botryosphaeriaceae species are known causal agents of grapevine trunk disease worldwide (3). References: (1) P. W. Crous et al. Stud. Mycol. 55:235, 2006. (2) B. Slippers et al. Mycologia 96:83, 2004. (3) J. R. Urbez-Torres. Phytopathol. Mediterr. 50:S5, 2011.

scholarly journals Effect of Temperature on Conidial Germination of Botryosphaeriaceae Species Infecting Grapevines

Plant Disease ◽  
2010 ◽  
Vol 94 (12) ◽  
pp. 1476-1484 ◽  
Author(s):  
José R. Úrbez-Torres ◽  
Emilie Bruez ◽  
José Hurtado ◽  
Walter D. Gubler
Keyword(s):  
Incubation Time ◽  
Conidial Germination ◽  
Effect Of Temperature ◽  
Botryosphaeria Dothidea ◽  
Diplodia Seriata ◽  
Neofusicoccum Parvum ◽  
Lasiodiplodia Theobromae ◽  
Different Temperatures ◽  
Northern And Southern Hemispheres ◽  
Botryosphaeriaceae Species

Germination of conidia of eight botryosphaeriaceous fungi infecting grapevines was evaluated after 2, 4, 6, 12, and 24 h incubation under eight different temperatures (5, 10, 15, 20, 25, 30, 35, and 40°C). The effect of temperature on conidial germination was also evaluated in different stages (hyaline versus pigmented conidia) of the species Lasiodiplodia theobromae. Conidial germination of Botryosphaeriaceae species infecting grapevines was significantly affected by temperature. Overall, conidial germination increased significantly with longer incubation times, especially from 2 to 12 h. In most cases, germination of conidia was not significantly different between 12 and 24 h incubation. Conidia of botryosphaeriaceous species did not germinate (with the exception of Botryosphaeria dothidea and Neofusicoccum parvum) at 5°C, and only B. dothidea, Diplodia seriata, and L. theobromae showed high levels of conidial germination at 40°C. Optimum conidial germination temperatures (defined as the temperature in which germination reached at least 50% in the shortest incubation time) were 25°C for B. dothidea and Dothiorella iberica, 25 to 30°C for Spencermartinsia viticola, 30°C for Diplodia corticola, D. mutila, D. seriata, N. parvum, and hyaline L. theobromae, and 40°C for pigmented L. theobromae conidia. Successful conidial germination of species of Botryosphaeriaceae infecting grapevines was always observed between 10 and 35°C with the exception of Dothiorella iberica and pigmented L. theobromae conidia, neither of which germinated at 35 and 10°C, respectively. Results of this study show conidia of botryosphaeriaceous species infecting grapevines to be capable of germination under a broad range of temperatures including those considered to be extreme, which may explain the success of these species as grapevine pathogens throughout most of the grape-growing areas in both Northern and Southern Hemispheres.

scholarly journals First Report of Botryosphaeria dothidea, Diplodia seriata, and Neofusicoccum luteum Associated with Canker and Dieback of Grapevines in Tunisia

Plant Disease ◽  
2014 ◽  
Vol 98 (3) ◽  
pp. 420-420 ◽  
Author(s):  
S. Chebil ◽  
R. Fersi ◽  
A. Yakoub ◽  
S. Chenenaoui ◽  
M. Chattaoui ◽  
...  
Keyword(s):  
Morphological Characteristics ◽  
Aerial Mycelium ◽  
Its2 Region ◽  
Pathogenicity Test ◽  
Botryosphaeria Dothidea ◽  
Diplodia Seriata ◽  
First Report ◽  
Grapevine Dieback ◽  
Pinus Spp ◽  
Neofusicoccum Luteum

In 2011, common symptoms of grapevine dieback were frequently observed in 2- to 5-year-old table grape (Vitis vinifera L.) cvs. in four vineyards located in northern Tunisia. The symptoms included dead spur and cordons, shoot dieback, and sunken necrotic bark lesions, which progressed into the trunk resulting in the death of large sections of the vine. Longitudinal and transversal sections of cordons and spurs from symptomatic vines revealed brown wedge-shaped cankers of hard consistency. Twelve symptomatic samples from spur and cordons were collected, surface disinfected by dipping into 5% (v/v) sodium hypochlorite for 2 min, and small pieces from the edge of necrotic and healthy tissue were removed and plated onto potato dextrose agar (PDA) at 25°C in the dark. Based on colony and conidia morphological characteristics, isolates were divided in three species, named Diplodia seriata, Botryosphaeria dothidea, and Neofusicoccum luteum. D. seriata colonies were gray-brown with dense aerial mycelium producing brown cylindric to ellipsoid conidia rounded at both ends and averaged 22.4 × 11.7 μm (n = 50). B. dothidea colonies were initially white with abundant aerial mycelium, gradually becoming dark green olivaceous. Conidia were fusiform to fusiform elliptical with a subobtuse apex and averaged 24.8 × 4.7 μm (n = 50). N. luteum colonies were initially pale to colorless, gradually darkening with age and becoming gray to dark gray producing a yellow pigment that diffuses into the agar. Conidia were hyaline, thin-walled, aseptate, fusiform to fusiform elliptical, and averaged 19.8 × 5.5 μm (n = 50). Identity of the different taxa was confirmed by sequence analyses of the internal transcribed spacer (ITS1-5.8S-ITS2) region of the rDNA and part of the elongation factor 1-alpha (EF1-α) gene. BLAST analysis of sequences indicated that six isolates were identified as D. seriata (GenBank: AY259094, AY343353), one isolate as B. dothidea (AY236949, AY786319) and one isolate as N. luteum (AY259091, AY573217). Sequences were deposited in GenBank under accessions from KC178817 to KC178824 and from KF546829 to KF546836 for ITS region and EF1-α gene, respectively. A pathogenicity test was conducted on detached green shoots cv. Italia for the eight Botryosphaeriaceae isolates. Shoots were inoculated by placing a colonized agar plug (5 mm diameter) from the margin of a 7-day-old colony on fresh wound sites made with a sterilized scalpel. Each wound was covered with moisturized cotton and sealed with Parafilm. Control shoots were inoculated using non-colonized PDA plugs. After 6 weeks, discoloration of xylem and phloem and necrosis with average length of 38.8, 17.6, and 11.2 mm were observed from inoculated shoots with D. seriata, N. luteum, and B. dothidea, respectively, and all three fungi were re-isolated from necrotic tissue, satisfying Koch's postulates. Control shoots showed no symptoms of the disease and no fungus was re-isolated. In Tunisia, Botryosphaeria-related dieback was reported only on citrus tree caused by B. ribis (2), on Pinus spp. caused by D. pinea (4), on Quercus spp. caused by D. corticola (3), and on olive tree (Olea europea) caused by D. seriata (1). To our knowledge, this is the first report of D. seriata, B. dothidea, and N. luteum associated with grapevine dieback in Tunisia. References: (1) M. Chattaoui et al. Plant Dis. 96:905, 2012. (2) H. S. Fawcett. Calif. Citrogr. 16:208, 1931. (3) B. T. Linaldeddu et al. J. Plant Pathol. 91:234. 2009. (4) B. T. Linaldeddu et al. Phytopathol. Mediterr. 47:258, 2008.

scholarly journals Antifungal Agents Based on Chitosan Oligomers, ε-polylysine and Streptomyces spp. Secondary Metabolites against Three Botryosphaeriaceae Species

Antibiotics ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 99 ◽  
Author(s):  
Laura Buzón-Durán ◽  
Jesús Martín-Gil ◽  
Eduardo Pérez-Lebeña ◽  
David Ruano-Rosa ◽  
José L. Revuelta ◽  
...  
Keyword(s):  
Cost Effective ◽  
Botryosphaeria Dothidea ◽  
Diplodia Seriata ◽  
Neofusicoccum Parvum ◽  
Grapevine Trunk Diseases ◽  
Streptomyces Spp ◽  
Effective Option ◽  
Trunk Diseases ◽  
Botryosphaeriaceae Species

Grapevine trunk diseases (GTDs) are a major threat to the wine and grape industry. The aim of the study was to investigate the antifungal activity against Neofusicoccum parvum, Diplodia seriata, and Botryosphaeria dothidea of ε-polylysine, chitosan oligomers, their conjugates, Streptomyces rochei and S. lavendofoliae culture filtrates, and their binary mixtures with chitosan oligomers. In vitro mycelial growth inhibition tests suggest that the efficacy of these treatments, in particular those based on ε-polylysine and ε-polylysine:chitosan oligomers 1:1 w/w conjugate, against the three Botryosphaeriaceae species would be comparable to or higher than that of conventional synthetic fungicides. In the case of ε-polylysine, EC90 values as low as 227, 26.9, and 22.5 µg·mL−1 were obtained for N. parvum, D. seriata, and B. dothidea, respectively. Although the efficacy of the conjugate was slightly lower, with EC90 values of 507.5, 580.2, and 497.4 µg·mL−1, respectively, it may represent a more cost-effective option to the utilization of pure ε-polylysine. The proposed treatments may offer a viable and sustainable alternative for controlling GTDs.

scholarly journals First Report of Gummosis Disease of Japanese Apricot Caused by Botryosphaeria dothidea in Taiwan

Plant Disease ◽  
2011 ◽  
Vol 95 (1) ◽  
pp. 77-77
Author(s):  
Y. Ko ◽  
C. W. Liu ◽  
S. S. Chen ◽  
K. Y. Chiu ◽  
Y. W. Sun ◽  
...  
Keyword(s):  
Direct Sequencing ◽  
Morphological Characteristics ◽  
Pcr Amplification ◽  
Conidial Suspension ◽  
Botryosphaeria Dothidea ◽  
Cortical Cells ◽  
Japanese Apricot ◽  
First Report ◽  
Representative Sequence ◽  
Plastic Bags

Japanese apricot (Prunus mume Sieb. et Zucc.) is an economically important fruit crop grown on more than 10,000 ha in Taiwan. During May 2008, twigs of Japanese apricot trees in the commercial farms of Renai Region (Nantou County) showed symptoms of gummosis disease, with 12 to 18% of the trees affected. The disease was more severe on trees weakened by drought stress. Limb and twig infections began around lenticles as small, sunken, discolored lesions at the margins of wounds. Following infection, cortical cells collapsed, bark became depressed, and blisters developed, which were often cracked with whitish gummy exudation. Necrotic areas were seen on the cortical tissues. Leaves showed yellowing and drooping. In winter months, numerous black pycnidia or perithecia formed on infected twigs. Single conidial isolates of the pathogen were obtained from diseased twigs on acidified potato dextrose agar (PDA) incubated at 25 ± 1°C for 3 days. On the basis of morphological characteristics, the fungus was identified as Botryosphaeria dothidea (3). Conidia (17 to 22.6 × 4.3 to 6.0 μm) were hyaline, unicellular, and spindle shaped. Asci (78 to 125 × 15 to 17 μm) were hyaline, bitunicate, clavate, and eight spored. Ascospores (18 to 22 × 7.0 to 8.2 μm) were hyaline and spindle shaped or fusoid. The pathogen identity was further confirmed by PCR amplification and sequencing of ribosomal DNA internal transcribed spacer from the fungus with the primers ITS5: 5′-GGAAGTAAAAGTCGTAACAAGG-3′ and ITS4: 5′-TCCTCCGCTTATTGATATGC-3′ (4), and a representative sequence was deposited in NCBI GenBank (Accession No. GU594225). The sequence showed 99 to 100% homology with previously characterized strains of B. dothidea (GenBank Accession Nos. EU441944, DQ177876, and AY786320). Pathogenicity tests were conducted with inoculum prepared by culturing the fungus on PDA under a continuous photoperiod of 128 ± 25 μE·m–2·s–1 at 25°C for 3 days. Shallow cuts (3 × 3 × 3 mm) were made on 12- to 15-month-old healthy twigs with a scalpel and inoculated with either a 5-mm mycelial disc or 0.5 ml of conidial suspension (105 conidia/ml) of the fungus. Two twigs on each of six trees were inoculated. Inoculated areas were covered with moist, sterile cotton and the entire twigs were enclosed in plastic bags. Twigs were inoculated with 5-mm PDA discs or sterile water for controls. The symptoms described above were observed on all inoculated twigs 14 days after inoculation, whereas control twigs remained healthy. Reisolation from the inoculated twigs consistently yielded B. dothidea. In Taiwan, B. dothidea has been reported as the causal agent of gummosis of peach (1) and fruit ring rot of pear (2); however, to our knowledge, this is the first report of B. dothidea causing gummosis on Japanese apricot. References: (1) Y. Ko et al. Plant Pathol. Bull. 1:70, 1992. (2) Y. Ko et al. Plant Prot. Bull. (Taiwan) 35:211, 1993. (3) B. Slippers et al. Mycologia 96:83, 2004. (4) T. J. White et al. In: Amplification and Direct Sequencing of Fungal Ribosomal RNA Genes for Phylogenetics. Academic Press. San Diego, CA, 1990.

scholarly journals First Report of Lasiodiplodia theobromae Associated with Stem Canker of Cassia fistula in Guangxi, South China

Plant Disease ◽  
2015 ◽  
Vol 99 (2) ◽  
pp. 288-288 ◽  
Author(s):  
T. J. Deng ◽  
Q. L. Li ◽  
X. L. Chen ◽  
S. P. Huang ◽  
T. X. Guo ◽  
...  
Keyword(s):  
Ornamental Plants ◽  
Morphological Characteristics ◽  
Its Region ◽  
Stem Canker ◽  
Morphological Identification ◽  
Vegetable Crops ◽  
Cassia Fistula ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Surface Sterilization

Cassia fistula, a member of the Fabaceae, known as the golden shower tree, is native to South Asia. It is now distributed worldwide and is popular as an ornamental plant as well as being used in herbal medicine. In October 2013, symptoms of stem canker were observed on C. fistula in a nursery (108°38′ E, 22°87′ N) in Nanning, Guangxi, China. The symptoms began as small brown lesions, which enlarged over several months to long, striped, slightly sunken lesions, 1 to 9 cm in width and 16 to 135 cm in length. The conspicuous cankers had vertical cracks outlining the canker and evenly spaced horizontal cracks, eventually resulting in whole plants dying back. The cankers were found on 90% of six-year-old plants in this nursery and were also observed in other plantings. On potato dextrose agar (PDA), isolates with similar morphological characteristics were consistently recovered from symptomatic plant tissues after surface sterilization in 75% ethanol for 30 sec and then in 0.1% mercuric chloride for 2 min. Over 100 conidia were examined from three isolates and were found to be elliptical and hyaline when immature, becoming dark brown, one-septate, and longitudinally striate when mature and ranging from 20 to 31 × 11 to 16 μm (average 25.5 × 13.6 μm). The rDNA internal transcribed spacer (ITS) region of isolate LC-1 was sequenced (GenBank Accession No. KM387285), and it showed 100% identity to Lasiodiplodia theobromae (Pat.) Griffon & Maubl. (GenBank KC964548), confirming the morphological identification (2) as L. theobromae (also known as Botryosphaeria rhodina (Cooke) Arx). A culture of this isolate has been preserved in the Guangxi Academy of Agricultural Sciences fungal collection. The pathogenicity of the isolate was tested on healthy twigs and branches of C. fistula trees in a field setting at Guangxi Agricultural Vocational-Technical College, Nanning, Guangxi, in June and August 2014. For each treatment, five green twigs and five 2-year-old branches were used. Five adjacent needle punctures were made on each branch with a sterilized needle. A mycelial plug was then placed on the wound of each branch and wrapped with Parafilm. Control twigs were treated with sterile PDA plugs. One week later, typical lesions were observed on the inoculated branches, with symptoms becoming more extensive after two weeks, but no symptoms were seen on the controls. Koch's postulates were fulfilled by re-isolation of L. theobromae from diseased branches. L. theobromae is recognized as an important wood pathogen and has been reported to cause cankers, dieback, and fruit and root rots in over 500 different hosts, including perennial fruit and nut trees, vegetable crops, and ornamental plants (2). The fungus has been reported on C. fistula in India since the 1970s (1); however, to our knowledge, this is the first report of L. theobromae infecting C. fistula in China. References: (1) R. S. Mathur. The Coelomycetes of India. Bishen Singh Mahendra Pal Singh, Delhi, India, 1979. (2) J. R. Úrbez-Torres et al. Plant Dis. 92:519, 2008.

scholarly journals First Report of a Canker Disease of Walnut Caused by Botryodiplodia theobromae in Egypt

Plant Disease ◽  
2007 ◽  
Vol 91 (2) ◽  
pp. 226-226 ◽  
Author(s):  
W. M. Haggag ◽  
M. S. M. Abou Rayya ◽  
N. E. Kasim
Keyword(s):  
Morphological Characteristics ◽  
Fungal Growth ◽  
Research Centre ◽  
Botryodiplodia Theobromae ◽  
Outer Bark ◽  
Canker Disease ◽  
First Report ◽  
Plant Foliage ◽  
Affected Tissue ◽  
And Control

Botryodiplodia spp. are known to produce cankers and dieback of several woody hosts. Botryodiplodia diseases were observed in 7-year-old orchards in Rhafah, north of Sinai, Egypt, in July 2005 and 2006. Symptoms appeared as dieback and cankers with dead leaves that were covered mostly with grayish white fungal growth; black pycnidia appeared on the surface of the infected branches. Plant foliage was discolored and partially or completely dry. When the outer bark was removed, the affected tissue appeared dark brown, in contrast to the yellowish green of healthy inner bark. On the basis of morphological characteristics (3), these fungi were identified as Botryodiplodia theobromae Pat. by the Plant Pathology Department, National Research Centre. Sporulating lesions were black and had a rough surface caused by the erumpent, confluent arrangement of pycnidia formed in infected tissue. The pycnidia were smallest in naturally infected twigs in nutritionally rich medium such as oatmeal agar (190 to 887 × 155 to 705 μm). Conidia were initially hyaline and unicellular, subovoid to ellipsoidal with a granular content. Mature conidia were two-celled, cinnamon to light brown, and often with longitudinal striations. Conidia measured 20 to 30 × 12 to 15 μm. Pathogenicity of isolates from symptomatic branches was determined by branch inoculations on rooted cuttings made from 7-year-old walnut trees growing in plastic pots. One isolate was inoculated on wounded and unwounded twigs using 100 μl of a suspension of 5 × 105 conidia per ml. Control branches were sprayed with water. All inoculated and control plants were kept in a greenhouse and watered as needed. There were three replicate plants for each isolate and inoculation technique that was used. After 3 weeks, cankers and grayish necrotic lesions developed on all inoculated plants. Samples of tissue from 10 infected walnut branches were plated on water agar. B. theobromae was recovered from all sampled plants. Control plants did not display any symptoms. B. theobromae has been reported on species of walnut from the Hermosillo Coast of Mexico (1) and India (2). To our knowledge, this is the first report of walnut dieback and canker caused by this pathogen in Egypt. References: (1) J. A. Arredondo. Rev. Mex. Fitopatol. 12:138, 1994. (2) B. Kusum. Indian J. Mycol. Plant Pathol. 21:295, 1991. (3). S. Masilamani and J. Muthumary. Mycol. Res. 100:1383, 1996.

scholarly journals First Report of Dieback Disease on Cedars Caused by Diplodia seriata in China

Plant Disease ◽  
2014 ◽  
Vol 98 (9) ◽  
pp. 1279-1279 ◽  
Author(s):  
Z. J. Jiao ◽  
Y. C. Kan ◽  
S. L. Huang
Keyword(s):  
Morphological Characteristics ◽  
Its Sequences ◽  
Fluorescent Light ◽  
Foeniculum Vulgare ◽  
Diplodia Seriata ◽  
First Report ◽  
Rdna Its ◽  
Rdna Its Sequences ◽  
Dieback Disease ◽  
Pinus Spp

Cedars (Cedrus deodara (Roxb.) G. Don) are well known as evergreen ornamental trees widely used in horticulture in temperate climates. In March 2013, dieback symptoms were found on cedar trees in different locations (including the campus of Nanyang Normal University) in Nanyang (33°01′ N, 112°29′ E), a southwestern city of Henan Province, China. Characteristic symptoms included needle discoloration and defoliation, canker formation and gummosis on trunks and branches, browning and tissue necrosis under the bark as well as dieback of branches/trunks. Of 873 cedar trees investigated, 139 (16%) were symptomatic. A total of 21 Diplodia sp. isolates were obtained from 102 tissue pieces randomly sampled from the lesion margins of 31 affected trees with a conventional method for isolation of culturable fungal species from plant tissues. Monohyphal cultures were isolated from actively growing edges of colonies to purify the isolates. The purified isolates were grown on 2% water agar with sterilized stems of Foeniculum vulgare to induce their colonies to form pycnidia (4). Unmatured conidia were hyaline, aseptate, and turned light to dark brown with maturity. Mature conidia were aseptate (rarely uniseptate), ovoid with truncated or rounded base and obtuse apex, externally smooth, roughened on the inner surface, and 8 to 11 × 23 to 26 μm (n = 50). These morphological characteristics of the isolates agreed with those of Diplodia seriata (the anamorph of Botryosphaeria obtusa) (5). The rDNA-ITS sequences of two representative isolates (xs-01 and xs-06) were amplified with primers ITS1 and ITS4. PCR products were purified and ligated with PMD-19T vector for sequencing. The rDNA-ITS sequences were submitted to GenBank with accession nos. KJ463386 and KJ549774 for isolates xs-01 and xs-06, respectively, showing 100% identity with multiple isolates of D. seriata (HQ660463, KC461297, and KF535906). Koch's postulates were fulfilled in greenhouse tests on 2-year-old cedar plantlets inoculated by the two isolates. Ten plantlets were used for the inoculation tests for each of the isolates, and their trunks were wounded to a 2 mm depth with a sterilized cork borer (3 mm diameter). The wounds were inoculated by mycelial plugs cut from 7-day-old colonies grown on potato dextrose agar (PDA) plates and wrapped with Parafilm, and those inoculated with pure PDA plugs served as control. Inoculated plantlets were incubated in a greenhouse with alternating cycles of 14 h fluorescent light/10 h darkness under moist conditions for 30 to 60 days at 28°C. Nine of 20 inoculated plantlets developed needle discoloration and shoot blight symptoms similar to those observed on naturally infected cedar trees. The control plantlets remained symptomless during the incubation period. D. seriata cultures were constantly recovered from each diseased plantlet, indicating that the isolated D. seriata isolates were responsible for the disease. D. seriata has been reported as a phytopathogen causing dieback diseases worldwide on multiple woody plant species such as olive (4), mulberry (1), Pinus spp., and Picea glauca (2,3). To our knowledge, this is the first report of D. seriata causing dieback disease on cedars in China. References: (1) M. Arzanlou et al. Arch. Phytopathol. Plant Protect. 46:682, 2013. (2) T. Burgess et al. Appl. Environ. Microbiol. 67:354, 2001. (3) G. Hausner et al. Can. J. Plant Pathol. 21:256, 1999. (4) J. Kaliterna et al. Plant Dis. 96:290, 2012. (5) A. J. L. Phillips et al. Fungal Divers. 25:141, 2007.

scholarly journals First Report of Neofusicoccum parvum Causing Panicle Blight and Leaf Spot on Vitis heyneana in China

Plant Disease ◽  
2015 ◽  
Vol 99 (3) ◽  
pp. 417-417 ◽  
Author(s):  
D. D. Wu ◽  
G. Fu ◽  
Y. F. Ye ◽  
F. Y. Hu ◽  
H. F. Mou ◽  
...  
Keyword(s):  
Leaf Spot ◽  
Severe Disease ◽  
Morphological Characteristics ◽  
Aerial Mycelium ◽  
Its Region ◽  
Average Diameter ◽  
Wine Production ◽  
Neofusicoccum Parvum ◽  
First Report ◽  
Panicle Blight

The climbing vine, Vitis heyneana Roem. & Schult, is a member of the grape family endemic to Asia. Its fruits are used in wine production, and its roots, stems, and leaves can be used in medicinal materials. This plant is grown in Southwest China, as well as in India, Bhutan, and Nepal. Mulao Autonomous County in Guangxi Province is the only artificial cultivation area in China. During the summer of 2013, a panicle blight and leaf spot were detected on V. heyneana on four farms in Mulao Autonomous County. The symptoms were observed from the onset of florescence through fruit harvest. Brown lesions initially appeared at the base of a panicle and then extended to the whole panicle, finally causing the panicle to die and fruit to drop. When the disease developed on leaves, the symptom initially appeared as small dark brown circular spots, later enlarging into irregular spots (average diameter 6 mm) with a light brown center and dark brown rim. With severe disease, some individual leaves were affected by numerous spots, leading to premature senescence. Small sections of diseased tissue excised from 10 panicle and 10 leaf samples were plated on potato dextrose agar (PDA) and incubated at 28°C. Fungal colonies developed, initially with abundant white aerial mycelium, which turned olivaceous gray after 5 days and formed black pycnidia after 25 days. The conidia were hyaline, ellipsoidal to fusiform, externally smooth, thin-walled, and nonseptate. Thirty conidia were measured; the dimensions were 12.0 to 17.5 × 4.0 to 6.0 μm. Morphological characteristics of the isolates were similar to the descriptions of Neofusicoccum parvum (3). The isolate MPT-1 was selected as a representative for molecular identification. Genomic DNA was extracted and used for PCR to amplify the internal transcribed spacer (ITS) region and partial translation elongation factor 1-alpha (EF1-α) gene, using primers ITS1/ITS4 and EF1-728F/EF1-986R, respectively. The obtained ITS sequence (GenBank Accession No. KJ599627) and EF1-α sequence (KM921768) showed >99% homology with several GenBank sequences of N. parvum. Morphological and molecular results confirmed the isolate as N. parvum. For pathogenicity tests, detached, young healthy panicles and leaves of V. heyneana were surface-sterilized, wounded by sterile needle, and inoculated with mycelial plugs (3 mm in diameter) of four N. parvum isolates. Ten panicles and 10 leaves were used for every isolate. Control panicles and leaves were treated with sterile PDA plugs. All the samples were placed in a humid chamber (RH 90%, 28°C, 12 h of light) for 3 days. Symptoms similar to those observed in the field developed on all panicles and leaves inoculated with N. parvum isolates. N. parvum was reisolated from all inoculated, symptomatic tissues. The controls remained symptomless. N. parvum has been reported to cause trunk canker on V. vinifera (2), dieback on Cupressus funebris (3), and a leaf spot on Myristica fragrans (1). To our knowledge, this is the first report of N. parvum causing panicle blight and leaf spot on V. heyneana in China. Panicle blight caused a large number of fruits to drop and reduced the yield seriously. Some effective measures should be taken to control this disease. References: (1) V. Jayakumar et al. New Dis. Rep. 23:19, 2011. (2) J. Kaliternam et al. Plant Dis. 97:1656, 2013. (3) S. B. Li et al. Plant Dis. 94:641, 2010.

scholarly journals First Report of Lasiodiplodia theobromae Causing Postharvest Fruit Rot on Guava (Psidium guajava) in Malaysia

Plant Disease ◽  
2021 ◽  
Author(s):  
Kar Yan Zee ◽  
Norhayu Asib ◽  
Siti Izera Ismail
Keyword(s):  
Morphological Characteristics ◽  
Its Region ◽  
Psidium Guajava ◽  
Fruit Rot ◽  
Pathogenicity Test ◽  
Tropical Fruit ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Guava Fruit ◽  
Initial Symptoms

Guava (Psidium guajava L.) is an economically important tropical fruit crop and is cultivated extensively in Malaysia. In September and October 2019, postharvest fruit rot symptoms were observed on 30% to 40% of guava fruit cv. Kampuchea in fruit markets of Puchong and Ipoh cities in the states of Selangor and Perak, Malaysia. Initial symptoms appeared as brown, irregular, water-soaked lesions on the upper portion of the fruit where it was attached to the peduncle. Subsequently, lesions then progressed to cover the whole fruit (Fig.1A). Lesions were covered with an abundance of black pycnidia and grayish mycelium. Ten symptomatic guava fruit were randomly collected from two local markets for our investigation. For fungal isolation, small fragments (5×5 mm) were excised from the lesion margin, surface sterilized with 0.5% NaOCl for 2 min, rinsed three times with sterile distilled water, placed on potato dextrose agar (PDA) and incubated at 25 °C with 12-h photoperiod for 2-3 days. Eight single-spore isolates with similar morphological characteristics were obtained and two representative isolates (P8 and S9) were characterized in depth. Colonies on PDA were initially composed of grayish-white aerial mycelium, but turned dark-gray after 7 days (Fig. 1B). Abundant black pycnidia were observed after incubation for 4 weeks. Immature conidia were hyaline, aseptate, ellipsoid, thick-walled, and mature conidia becoming dark brown and 1-septate with longitudinal striations, 25.0 − 27.0 ± 2.5 × 13.0 − 14.0 ± 1.0 μm (n = 30) (Fig.1C, D). On the basis of morphology, both representative isolates were identified as Lasiodiplodia theobromae (Pat.) Griffon & Maubl. (Alves et al. 2008). For molecular identification, genomic DNA of the two isolates was extracted using the DNeasy plant mini kit (Qiagen, USA). The internal transcribed spacer (ITS) region of rDNA and translation elongation factor 1-alpha (EF1-α) genes were amplified using ITS5/ITS4 and EF1-728F/EF1-986R primer set, respectively (White et al. 1990, Carbone and Kohn 1999). BLASTn analysis of the resulting ITS and EF1-α sequences indicated 100% identity to L. theobromae ex-type strain CBS 164.96 (GenBank accession nos: AY640255 and AY640258, respectively) (Phillips et al. 2013). The ITS (MW380428, MW380429) and EF1-α (MW387153, MW387154) sequences were deposited in GenBank. Phylogenetic analysis using the maximum likelihood based on the combined ITS-TEF sequences indicated that the isolates formed a strongly supported clade (100% bootstrap value) to the related L. theobromae (Kumar et al. 2016) (Fig.2). A pathogenicity test of two isolates was conducted on six healthy detached guava fruits per isolate. The fruit were surface sterilized using 70% ethanol and rinsed twice with sterile water prior inoculation. The fruit were wound-inoculated using a sterile needle according to the method of de Oliveira et al. (2014) and five-mm-diameter mycelial agar plugs from 7-days-old PDA culture of the isolates were placed onto the wounds. Six additional fruit were wound inoculated using sterile 5-mm-diameter PDA agar plugs to serve as controls. Inoculated fruit were placed in sterilized plastic container and incubated in a growth chamber at 25 ± 1 °C, 90% relative humidity with a photoperiod of 12-h. The experiment was conducted twice. Five days after inoculation, symptoms as described above developed on the inoculated sites and caused a fruit rot, while control treatment remained asymptomatic. L. theobromae was reisolated from all symptomatic tissues and confirmed by morphological characteristics and confirmed by PCR using ITS region. L. theobromae has recently been reported to cause fruit rot on rockmelon in Thailand (Suwannarach et al. 2020). To our knowledge, this is the first report of L. theobromae causing postharvest fruit rot on guava in Malaysia. The occurrence of this disease needs to be monitored as this disease can reduce the marketable yield of guava. Preventive strategies need to be developed in the field to reduce postharvest losses.

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