scholarly journals First Report of Lasiodiplodia theobromae Associated with Decline of Grapevine Rootstock Mother Plants in Spain

Plant Disease ◽  
2008 ◽  
Vol 92 (5) ◽  
pp. 832-832 ◽  
Author(s):  
A. Aroca ◽  
R. Raposo ◽  
D. Gramaje ◽  
J. Armengol ◽  
S. Martos ◽  
...  
Keyword(s):  
Elongation Factor ◽  
Aerial Mycelium ◽  
Its Region ◽  
Pathogenic Fungi ◽  
Vascular Lesions ◽  
Malt Extract ◽  
Pathogenicity Test ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Mother Plants

A field of Richter 110 rootstock mother plants in Valencia Province (eastern Spain) was surveyed during November 2006 to study the mycoflora of declining plants. Two canes with stunted leaves were collected from a plant with a reduced number of shoots. No cankers or vascular lesions were observed in the collected canes. Six wood chips (1 to 2 mm thick) were taken from one basal fragment (3 to 4 cm long) of each cane, surface sterilized in 70% ethanol for 1 min, and plated on malt extract agar supplemented with 0.5 g L–1 of streptomycin sulfate. Petri dishes were incubated for 7 days at 25°C. A fungus was consistently isolated from all samples that showed the following characteristics: colonies grown on potato dextrose agar (PDA) at 25°C developed a white, aerial mycelium that turned gray after 4 to 6 days and produced pycnidia after 1 month on sterile grapevine slivers of twigs placed on the PDA surface; conidia from culture were ellipsoidal, thick walled, initially hyaline, nonseptate, and measuring 20 to 25 (22.5) × 12 to 14 (13) μm; aged conidia were brown, 1-septate with longitudinal striations in the wall; and pseudoparaphyses variable in form and length were interspersed within the fertile tissue. The fungus was identified as Lasiodiplodia theobromae (Pat.) Griffon & Maubl. from the above characteristics (2). Identity was confirmed by analysis of the nucleotide sequences of the internal transcribed spacer (ITS) region from the rRNA repeat and part of the translation elongation factor 1-alpha (EF1-α) and the β-tubulin (B-tub) genes, as done elsewhere (1,3). BLAST searches at GenBank showed a high identity with reference sequences (ITS: 100%, EF1-α: 97%; B-tub: 99%). Representative sequences of the studied DNA regions were deposited at GenBank (Accession Nos.: ITS: EU254718; EF1-α: EU254719; and B-tub: EU254720). A pathogenicity test was conducted on 1-year-old grapevine plants cv. Macabeo grafted onto Richter 110 rootstocks maintained in a greenhouse. A superficial wound was made on the bark of 10 plants with a sterilized scalpel, ≈10 cm above the graft union. A mycelial plug obtained from the margin of an actively growing fungal colony (isolate JL664) was placed in the wound and the wound was wrapped with Parafilm. Ten additional control plants were inoculated with sterile PDA plugs. All control plants grew normally, and the inoculation wound healed 3 months after inoculation. Plants inoculated with L. theobromae showed no foliar symptoms in the same period, but developed cankers variable in size surrounding the inoculation sites. Vascular necroses measuring 8.4 ± 1.5 cm (mean ± standard error) developed in the inoculated plants that were significantly longer than the controls (0.3 ± 0.2 cm). The pathogen was reisolated from all inoculated plants and no fungus was reisolated from the controls. These results confirmed the pathogenicity of L. theobromae to grapevine and points to a possible involvement of L. theobromae in the aetiology of grapevine decline as previously reported (3,4). To our knowledge, this is the first report of L. theobromae isolated from grapevine in Spain. References: (1) J. Luque et al. Mycologia 97:1111, 2005. (2) E. Punithalingam. No. 519 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1976. (3) J. R. Úrbez-Torres et al. Plant Dis. 90:1490, 2006. (4) J. M. van Niekerk et al. Phytopathol. Mediterr. 45(suppl.):S43, 2006.

scholarly journals First Report of Mulberry Root Rot Caused by Lasiodiplodia theobromae in China

Plant Disease ◽  
2014 ◽  
Vol 98 (11) ◽  
pp. 1581-1581 ◽  
Author(s):  
H.-H. Xie ◽  
J.-G. Wei ◽  
F. Liu ◽  
X.-H. Pan ◽  
X.-B. Yang
Keyword(s):  
Root Rot ◽  
Management Strategies ◽  
Its Region ◽  
Pathogenic Fungi ◽  
Fruit Trees ◽  
Morphological Characters ◽  
Pathogenicity Test ◽  
Total Production ◽  
Lasiodiplodia Theobromae ◽  
First Report

Mulberry (Morus alba L.) is an important cash crop and medicinal plant that has been cultivated for more than 5,000 years in China. The area of mulberry production in Guangxi Province is 45% of total production in China, with 1.3 million ha planted. In recent years, a mulberry root rot occurred in Heng County covering all the mulberry planting farms. Observations of 200 diseased plants were made. The xylem of infected roots first turned brown, and then became black followed by cortex rot. The xylem and cortex of infected roots were easily separated. The xylem of the stem of symptomatic plants was also brown and the bark was slightly darker than normal. Leaves of diseased plants turned yellow and wilted, but the wilted leaves remained on the affected branches for about 3 weeks. All affected branches and stem dried after a month. The affected area was 12,000 ha with incidences varying from 13 to 52%. About 8% of young mulberry trees died in severely infested orchards. The disease caused more than $3 million in losses within a year in Heng County alone. The causal fungus was isolated from xylem tissues of symptomatic roots of 62 mulberry plants with an isolation rate of 90%. Pathogenicity test was made by inoculating 5-month-old healthy mulberry plants with PDA plugs (5 × 5 mm) grown 5 days with viable mycelia of the fungus. Nine healthy plants were wounded on the roots with a sterile knife, and mycelial plugs of three Lasiodiplodia theobromae (Pat.) Griffon & Maubl isolates were placed on the wounds, covered with sterile moist cotton, and wrapped with Parafilm. Nine control plants were treated with PDA plugs. The test was repeated three times. All treated plants were kept in a greenhouse at ~28°C and 40% RH. After 3 days, the root xylem of inoculated plants turned brown and gradually became dark, similar to symptoms observed in the field. After 8 days, inoculated seedlings gradually wilted, and all the treated plants died after 11 days with leaves undetached. The fungus was re-isolated from all nine diseased plants and no symptoms were observed on the roots of control plants. The causal agent, of which conidia were dark brown, one-septate, thick walled, and ellipsoid with 4 or 6 vertical lines of dashes, 12.50 to 13.75 × 13.75 to 25.63 μm (n = 100), was identified as L. theobromae based on morphological characters described by Punithalingam (3) and sequences of the ITS region of rDNA using primers ITS1 and ITS4 and EF1-α using primers EF728F and EF986R. The ITS sequence (HG917932) was similar to the ITS sequences of AY640255 (CBS164.96) and AY236952 (CMW9074) in GenBank with identities of 98.8 and 99.8%, respectively. The EF1-α sequence HG917934 was similar to that of AY640258 (CBS164.96) and AY236901 (CMW9074) with identities of 99.7 and 99.7%, respectively. L. theobromae is a cosmopolitan fungus causing both field and storage diseases on more than 280 plant species including crops, fruits, and cash fruit trees (1,2,5). Mulberry root rot caused by L. theobromae has been reported in India (4) and ours is the first report in China. This finding clarifies the pathogen of mulberry root rot previously thought as Fusarium sp. in China, which is critical to develop management strategies to control this disease. References: (1) N. M. Celiker and T. J. Michailides. New Dis. Rep. 25:12, 2012. (2) I. H. Fischer et al. Australia Plant Dis. Notes 3:116, 2008. (3) E. Punithalingam. Botryodiplodia theobromae. CMI Descriptions of Pathogenic Fungi and Bacteria No. 519. CAB International, Wallingford, UK, 1976. (4) N. V. Radhakrishnan et al. Indian Phytopathol. 48:490, 1995. (5) B. C. Sutton. The Coelomycetes. Commonwealth Mycology Institute, Kew, Surrey, England, 1980.

scholarly journals First Report of Euphorbia larica Dieback Caused by Fusarium brachygibbosum in Oman

Plant Disease ◽  
2013 ◽  
Vol 97 (5) ◽  
pp. 687-687 ◽  
Author(s):  
I. H. Al-Mahmooli ◽  
Y. S. Al-Bahri ◽  
A. M. Al-Sadi ◽  
M. L. Deadman
Keyword(s):  
Elongation Factor ◽  
Aerial Mycelium ◽  
Its Region ◽  
Common Component ◽  
First Report ◽  
Ethnobotanical Uses ◽  
Artificial Inoculations ◽  
Stem Lesions ◽  
Desert Flora

Euphorbia larica Boiss. (Arabic = Isbaq) is a dominant and common component of the native desert flora of northern Oman. Traditional ethnobotanical uses have included use of the latex for treating camels with parasites. In February 2011, E. larica plants showing stem lesions up to several cm long and in many cases with stem dieback were collected from Al-Khoudh 50 km west of Muscat. The disease appeared widespread within the location where several dead specimens were also recorded, although the cause was unclear. Sections (5 mm) of five diseased branches taken from different plants and placed on potato dextrose agar (PDA) in all cases yielded Fusarium-like colonies. Colonies recovered were initially white becoming rose to medium red in color with abundant aerial mycelium. Macroconidia were scarce and scattered (mean of 20 spores: 26.83 × 4.73 μm) with three to four septa per spore; microconidia were slightly curved, ovoid, and fusiform (mean of 20 spores: 11.64 × 4.03 μm) with zero to two septa per spore. Spherical chlamydospores (mean of 20 spores: 11.05 μm) were terminal and intercalary, single, and in chains. In vitro characters and spores measurements conformed to previously described features of Fusarium brachygibbosum Padwick (1). Mycelial plugs (5 mm) were taken from 7-day-old cultures of the fungus grown on 2.5% PDA and applied to a small incision (3 mm) on the stems of healthy E. larica grown in situ and protected with wet cotton and Parafilm. The residual agar, mycelium, cotton, and Parafilm were removed after 7 days and symptoms were recorded. Control stems were inoculated using PDA (5 mm) plugs alone and inoculations were repeated twice. Artificial inoculations resulted in dieback of all stems within 11 days and fungal colonies identical to initial isolations were recovered from artificially infected surface-sterilized stem pieces. Identification of F. brachygibbosum was confirmed by comparing sequences generated from the internal transcribed spacer (ITS) region of the ribosomal DNA (ITS1 and ITS4 primers) and the intron region of translation elongation factor alpha (EF1-α) (EF-1-986 and EF-728 primers). The ITS and EF1-α sequences were found to share 100% and 99% nucleotide similarity to previously published sequences of the ITS (HQ443206) and EF1-α (JQ429370) regions of F. brachygibbosum in GenBank. The accession number of ITS sequence of one isolate assigned to EMBL-Bank was HF562936. The EF sequence was assigned to EMBL-Bank accession (submission number Hx2000027017; number will be sent later). This pathogen has previously been reported on date palm (2) in Oman but, to our knowledge, this is the first report of this pathogen on E. larica. References: (1) A. M. Al-Sadi et al. Crop Prot. 37:1, 2012. (2) G. W. Padwick. Mycol. Pap. 12:11, 1945.

scholarly journals First Report of Vascular Wilt Caused by Fusarium redolens on Lentil in Italy

Plant Disease ◽  
2008 ◽  
Vol 92 (7) ◽  
pp. 1132-1132 ◽  
Author(s):  
L. Riccioni ◽  
A. Haegi ◽  
M. Valvassori
Keyword(s):  
Vascular Tissue ◽  
Fusarium Species ◽  
Elongation Factor ◽  
Morphological Characteristics ◽  
Its Region ◽  
Its Sequences ◽  
Homology Search ◽  
Control Treatment ◽  
Pathogenicity Test ◽  
First Report

Lentil (Lens culinaris Medik.) is a traditional crop in Sicily, Italy. Near Villalba (Caltanissetta), a local lentil landrace, “Lenticchia di Villalba”, is commonly grown. From 2002 to 2004, wilt was observed in five lentil fields (≈1 ha each) at rates from 5 to 20%. Affected plants were yellow and stunted with discoloration in the vascular tissue of stems and crowns. Pieces of brown vascular tissue from stems were disinfested in 2% sodium hypochlorite for 2 min, rinsed with sterile distilled water, placed on potato dextrose agar, and incubated at 23°C. Isolates with morphological characteristics of Fusarium oxysporum Schlecht.:Fr. (2) were consistently recovered from affected plants. For molecular identification of five isolates, the rDNA internal transcribed spacer (ITS) region and a portion of the elongation factor EF-1α were sequenced using ITS5/4 and EF1/2 primers, respectively (1). Two sequences of the ITS region were obtained: a 468-bp sequence from isolates ER1259, ER1260, and ER1275 (submitted as GenBank Accession No. EU159118) and a 483-bp sequence from isolates ER1274 and ER1276 (submitted as GenBank Accession No. EU281661). The two sequences shared 93% similarity. A sequence homology search using the NCBI BLAST program revealed that the first sequence had 100% homology with the ITS sequences of more than 50 F. oxysporum isolates of various formae speciales in GenBank and the second shared 100% homology with the ITS sequences of five isolates of F. redolens Wollenw. (e.g., GenBank Accession No. X94169 of the strain CBS 360.87). Amplification of the EF-1α produced a sequence from isolates ER1274 and ER1276 (submitted as GenBank Accession No. EU281660) with 99 to 100% homology to sequences of F. redolens and a sequence from strains ER1259, ER1275, and ER1260 (submitted as GenBank Accession No. EU281659) with 100% homology to that of more than 50 F. oxysporum strains in GenBank. Although F. redolens and F. oxysporum are morphologically similar, recent molecular studies have shown that they are distinct and phylogenetically distant species (3). On the basis of genetic sequences, isolates ER1274 and ER1276 were identified as F. redolens. These isolates were evaluated for pathogenicity on lentil. For each isolate, 10 2-week-old seedlings of “Lenticchia di Villalba” were inoculated by submerging roots in a suspension of 2.5 × 106 conidia/ml for 10 min. Plants were put into separate tubes containing 70 ml of a nutritional liquid medium (7 ml of HydroPlus Olikani per liter; Yara, Nanterre, France) and incubated in a growth chamber at 20°C with 12 h of light per day. Seedlings dipped in sterile water served as the control treatment. The pathogenicity test was repeated twice. Inoculated seedlings started to wilt 1 week after inoculation and developed root rot and vascular discoloration. After 2 weeks, 70% of the inoculated plants were affected by both isolates and 40 and 10% died when inoculated with ER1274 and ER1276 isolates, respectively. F. redolens was consistently reisolated from the stems of wilted plants. Noninoculated plants remained healthy. Currently, only F. oxysporum f. sp. lentis Vasud. and Sriniv. has been reported as the cause of Fusarium wilt of lentil. To our knowledge, this is the first report of F. redolens as a pathogen on lentil. References: (1) R. P. Baayen et al. Phytopathology 91:1037, 2001. (2) P. E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. The Pennsylvania State University Press, University Park, 1983. (3) K. O'Donnell et al. Mycologia 90:465, 1998.

scholarly journals First report of Fusarium falciforme (FSSC 3+4) causing root rot on chickpea in Mexico

Plant Disease ◽  
2021 ◽  
Author(s):  
Sixto Velarde Felix ◽  
Victor Valenzuela ◽  
Pedro Ortega ◽  
Gustavo Fierros ◽  
Pedro Rojas ◽  
...  
Keyword(s):  
Fusarium Solani ◽  
Root Rot ◽  
Species Complex ◽  
Elongation Factor ◽  
Morphological Characteristics ◽  
Aerial Mycelium ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Single Spore ◽  
First Report

Chickpea (Cicer aretinium L.) is a legume crop of great importance worldwide. In January 2019, wilting symptoms on chickpea (stunted grow, withered leaves, root rot and wilted plants) were observed in three fields of Culiacan Sinaloa Mexico, with an incidence of 3 to 5%. To identify the cause, eighty symptomatic chickpea plants were sampled. Tissue from roots was plated on potato dextrose agar (PDA) medium. Typical Fusarium spp. colonies were obtained from all root samples. Ten pure cultures were obtained by single-spore culturing (Ff01 to Ff10). On PDA the colonies were abundant with white aerial mycelium, hyphae were branched and septae and light purple pigmentation was observed in the center of old cultures (Leslie and Summerell 2006). From 10-day-old cultures grown on carnation leaf agar medium, macroconidias were falciform, hyaline, with slightly curved apexes, three to five septate, with well-developed foot cells and blunt apical cells, and measured 26.6 to 45.8 × 2.2 to 7.0 μm (n = 40). The microconidia (n = 40) were hyaline, one to two celled, produced in false heads that measured 7.4 to 20.1 (average 13.7) μm × 2.4 to 8.9 (average 5.3) μm (n = 40) at the tips of long monophialides, and were oval or reniform, with apexes rounded, 8.3 to 12.1 × 1.6 to 4.7 μm; chlamydospores were not evident. These characteristics fit those of the Fusarium solani (Mart.) Sacc. species complex, FSSC (Summerell et al. 2003). The internal transcribed spacer and the translation elongation factor 1 alpha (EF1-α) genes (O’Donnell et al. 1998) were amplified by polymerase chain reaction and sequenced from the isolate Ff02 and Ff08 (GenBank accession nos. KJ501093 and MN082369). Maximum likelihood analysis was carried out using the EF1-α sequences (KJ501093 and MN082369) from the Ff02 and Ff08 isolates and other species from the Fusarium solani species complex (FSSC). Phylogenetic analysis revealed the isolate most closely related with F. falciforme (100% bootstrap). For pathogenicity testing, a conidial suspension (1x106 conidia/ml) was prepared by harvesting spores from 10-days-old cultures on PDA. Twenty 2-week-old chickpea seedlings from two cultivars (P-2245 and WR-315) were inoculated by dipping roots into the conidial suspension for 20 min. The inoculated plants were transplanted into a 50-hole plastic tray containing sterilized soil and maintained in a growth chamber at 25°C, with a relative humidity of >80% and a 12-h/12-h light/dark cycle. After 8 days, the first root rot symptoms were observed on inoculating seedlings and the infected plants eventually died within 3 to 4 weeks after inoculation. No symptoms were observed plants inoculated with sterilized distilled water. The fungus was reisolated from symptomatic tissues of inoculated plants and was identified by sequencing the partial EF1-α gene again and was identified as F. falciforme (FSSC 3 + 4) (O’Donnell et al. 2008) based on its morphological characteristics, genetic analysis, and pathogenicity test, fulfilling Koch’s postulates. The molecular identification was confirmed via BLAST on the FusariumID and Fusarium MLST databases. Although FSSC has been previously reported causing root rot in chickpea in USA, Chile, Spain, Cuba, Iran, Poland, Israel, Pakistan and Brazil, to our knowledge this is the first report of root rot in chickpea caused by F. falciforme in Mexico. This is important for chickpea producers and chickpea breeding programs.

scholarly journals First Report of Chrysoporthe deuterocubensis Causing Canker on Syzygium samarangense in Taiwan

Plant Disease ◽  
2013 ◽  
Vol 97 (11) ◽  
pp. 1508-1508
Author(s):  
M. C. Fan ◽  
C. C. Huang ◽  
J. S. Huang ◽  
S. F. Tsai ◽  
H. C. Yeh ◽  
...  
Keyword(s):  
Apple Tree ◽  
Average Length ◽  
Its Region ◽  
Fruit Production ◽  
Molecular Characteristics ◽  
Malt Extract ◽  
Pathogenicity Test ◽  
Total Production ◽  
First Report ◽  
Β Tubulin

Wax apple (Syzygium samarangense Merr. & Perry, syn. Eugenia javanica Lam.) belongs to the Myrtaceae family is an important economical tree fruit in Taiwan. The total production acreage of wax apple was 5,266 ha in which more than 77% were located in Pingtung County, southern Taiwan, in 2012. Since the winter of 2010, symptoms of withering leaves and cracking branches on wax apple trees were observed in some orchards in Nanjhou and Linbian Townships, Pingtung County. Diseased trees declined gradually and resulted in reduced fruit production. On the bark of diseased twigs and branches, black conidiamata with yellowish orange conidia were usually observed. For diagnosis, tissues from symptomatic branches were excised, surface sterilized with 0.5% sodium hypochlorite, and placed on 2% water agar in petri dishes. A total of four identical fungal isolates were obtained and maintained on potato dextrose agar (PDA). To fulfill Koch's postulates, three twigs of a wax apple tree were wounded with scalpel and inoculated with each of the four isolates, one tree per isolate. A 7-day-old hyphal mat (about 7 × 18 mm) of each fungal isolate was attached on the wound, wrapped with a wet absorbent cotton and Parafilm, and then covered with a layer of aluminum foil. For the control, the twigs of a wax apple tree were inoculated with PDA plugs. The pathogenicity test was repeated once. After 30 days, withering leaves and cracking twigs were observed on inoculated twigs and the same pathogen was reisolated. Conversely, all of the non-inoculated plants remained healthy. Identification of the pathogen was conducted using its morphological, physiological, and molecular characteristics. On malt extract agar, the colony was floccose and white with hazel hues. The optimal temperature for the mycelial growth was 30°C. Conidia were hyaline, and oblong, with the average size of 4.7 ± 0.6 × 2.7 ± 0.2 μm (100 conidia). Ascostromata were semi-immersed in the bark with fusoid asci, eight ascospores per ascus. Ascospores were hyaline, 2-celled, and tapered in both ends, with the average length of 6.8 ± 0.7 × 2.4 ± 0.3 μm (100 ascospores). For molecular identification, the internal transcribed spacer (ITS) of ribosomal DNA and β-tubulin genes was amplified using the ITS1/ITS4 (3), Bt1a/Bt1b, and Bt2a/Bt2b (1) primer pairs. The gene sequences were deposited in GenBank (Accessions KC792616, KC792617, KC792618, and KC792619 for the ITS region; KC792620, KC792621, KC792622, and KC792623 for Bt1 region, and KC812732, KC812733, KC812734, and KC812735 for Bt2 region) and showed 99 to 100% identity to the Chrysoporthe deuterocubensis isolate CMW12745 (DQ368764 for ITS region; GQ290183 for Bt1 region, and DQ368781 for Bt2 region). In addition, the Bt1 region of the β-tubulin gene consisted of two restriction sites for AvaI and one restriction site for HindIII. This is identical to the description of C. deuterocubensis, a cryptic species in C. cubensis, by Van Der Merwe et al. (2). According to these results, the pathogen was identified as C. deuterocubensis Gryzenh. & M. J. Wingf. To the best of our knowledge, this is the first report of canker disease caused by C. deuterocubensis on S. samarangense in Taiwan. References: (1) N. L. Glass and G. C. Donaldson. Appl. Environ. Microbiol. 61:1323, 1995. (2) N. A. Van Der Merwe et al. Fungal Biol. 114:966, 2010. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

scholarly journals First report of panicle rot caused by Fusarium asiaticum on foxtail millet in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Fanxin Kong ◽  
Haijin Zhang ◽  
Zhi Liu ◽  
Guoqiu Chen ◽  
Jing Xu
Keyword(s):  
Foxtail Millet ◽  
Morphological Characteristics ◽  
Aerial Mycelium ◽  
Its Region ◽  
Liaoning Province ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Dark Cycle ◽  
First Report ◽  
Fusarium Asiaticum

Foxtail millet [ Setaria italica (L.) P. Beauv.] is one of the most important nutritious food crops. It is used for wine and health products in China. In August of 2019, panicle rot symptoms with up to 85% of panicles infected were observed on foxtail millet (cultivar Chaogu 8) in a commercial field located in Chaoyang city of Liaoning Province, China. Typical disease symptoms included brown spots on spikelets at early stages and brown-colored withering and rot of whole panicles at late stages, with the symptoms being more severe at the tip of the panicles. Under high humidity conditions, pink or salmon-colored molds developed on panicles. Symptomatic spikelet pieces were surface-disinfested with 70% ethanol for 1 min followed by 2% NaOCl for 3 min, rinsed with sterilized water for three times, and placed on potato dextrose agar (PDA) medium at 25°C. After 5 days, colonies turned pink to dark red with fluffy aerial mycelium and pigmentation with the age. Ten pure cultures were obtained from single conidia of mycelium grown on carnation leaf agar (CLA) medium at 25°C under a 12-h light-dark cycle using an inoculation needle under stereomicroscope. Macroconidia were hyaline, falcate with foot cells, 3–5 septate and size: 28.5- 44.0 μm × 3.8 - 4.9 μm. Chlamydospores were globose to subglobose (5.4 to 13.8 μm). No microconidia were produced on CLA. Black, ostiolate subglobose perithecia were formed on CLA after one month of incubation at 20°C under a 12-h light-dark cycle. Morphological characteristics of the fungus were in agreement with the description of Fusarium asiaticum (O’Donnell et al. 2004; Leslie and Summerell 2006). To validate this identification, partial translation elongation factor 1 alpha (TEF1-a) gene, and rDNA internal transcribed spacer (ITS) region of five isolates were amplified and sequenced (O’Donnell et al. 2015; White et al.1990). Identical sequences were obtained, and the sequence of one representative isolate (JGF-3) was submitted to GenBank. BLASTn analysis of both TEF sequence (MW685833) and ITS sequence (MW423687), revealed 100% sequence identity with F. asiaticum KT380120 and MT322117, respectively. Pathogenicity test were conducted on cultivar Chaogu 8 of foxtail millet. Inoculum was prepared from the culture of JGF-3 incubated in 2% mung beans juice on a shaker (140 rpm) at 25°C for 48 h. Conidial suspension (5 × 105 conidia per ml) was prepared and sprayed onto the panicles of 20 plants at the initial flowering stage and 20 additional plants that were sprayed with distilled water served as the non-inoculated controls. Treated plants were covered with plastic bags for 48 h and maintained at a greenhouse with day and night temperatures of 26 and 24°C, respectively. Two weeks after inoculation, all inoculated panicles exhibited symptoms similar to the syptoms observed in the field. No symptoms were observed in the non-inoculated control plants. The experiment was repeated twice with similar results. F. asiaticum was reisolated from the inoculated plants and its morphological characteristics matched those of the original isolates; the fungus was not reisolated from the non-inoculated plants. To our knowledge, this is the first report of F. asiaticum causing panicle rot of foxtail millet in China. To date, the disease has been observed to be present in Fuxin and Tieling city of Liaoning Province. Panicle rot can become an important disease in foxtail millet in China. References: O’Donnell, K., et al. 2004. Fungal Genetics and Biology 41: 600. Leslie, J. F., and Summerell, B. A. 2006. The Fusarium laboratory manual. Blackwell Publishing, Ames, pp 176-179. O’ Donnell, K., et al. 2015. Phytoparasitica 43: 583. White, T. J., et al. 1990. Academic Press, San Diego, CA, pp 315-322.

scholarly journals First Report of Choanephora Blight Caused by Choanephora infundibulifera on Hibiscus rosa-sinensis in Korea

Plant Disease ◽  
2014 ◽  
Vol 98 (9) ◽  
pp. 1275-1275 ◽  
Author(s):  
J. H. Park ◽  
S. E. Cho ◽  
K. S. Han ◽  
S. H. Lee ◽  
H. D. Shin
Keyword(s):  
Large Subunit ◽  
Aerial Mycelium ◽  
Its Region ◽  
The United States ◽  
Pathogenicity Test ◽  
Potential Threat ◽  
First Report ◽  
Hibiscus Rosa Sinensis ◽  
Initial Symptoms ◽  
Chinese Hibiscus

Hibiscus rosa-sinensis L., commonly known as Chinese hibiscus, is an evergreen flowering shrub belonging to the Malvaceae and is widely cultivated throughout Asia including Korea. In August 2013, blight was observed on Chinese hibiscus in a commercial flower nursery in Seoul, Korea. Initial symptoms began as reddish purple spots at the tip of flowers and expanded to encompass entire flowers. Infected lesions appeared water-soaked, reddish brown, and were followed by rapid rotting of infected tissues. Approximately 50% of the plants surveyed were affected. Monosporous sporangiola formed on infected tissue were transferred to potato dextrose agar (PDA). Fungal colonies were obtained that were at first white with abundant aerial mycelium, and then became yellowish with the appearance of sporangiola. Sporangiophores bearing sporangiola were erect to slightly curved, unbranched, and hyaline. Funnel-shaped secondary vesicles formed on the primary vesicles. Sporangiola were indehiscent, ovoid to subglobose, smooth, non-striated, brown to dark brown, 10 to 27.5 × 8.5 to 17 μm, and sometimes germinated in culture. The fungus was identified as Choanephora infundibulifera (Curr.) D.D. Cunn. based on the morphological and cultural characteristics (2). Voucher specimens were housed in the Korea University Herbarium (KUS). An isolate obtained from KUS-F27535 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC47643) and used for a pathogenicity test and molecular analyses. To confirm identity of the fungus, genomic DNA was extracted with DNeasy Plant Mini Kits (Qiagen Inc., Valencia, CA). The internal transcribed spacer (ITS) region of rDNA and the D1/D2 region of the large subunit (LSU) were amplified with the primers ITS1/ITS4 and NL1/LR3, respectively (3), and sequenced. The resulting 635-bp ITS and 680-bp D1/D2 sequences were deposited in GenBank (Accession Nos. KF486539 and KF486538). A GenBank BLAST search revealed that the ITS sequences showed 100% similarity with that of C. infundibulifera (JN943009) and D1/D2 sequences also showed 100% identity with that of C. infundibulifera (JN939193). A sporangiola suspension (2 × 104 cells/ml) was sprayed over three pots of the shrub, kept in a humid chamber for 2 days, and placed in greenhouse (28°C and 80 to 100% RH). Another three potted plants of the same age were sprayed with sterile water and served as controls. After 4 days, typical blossom blight symptoms, identical to the ones observed in the nursery, developed on the inoculated flowers. No symptoms were observed on controls. C. infundibulifera was re-isolated from inoculated plants. Pathogenicity test was conducted twice with the same results, fulfilling Koch's postulates. Choanephora blight caused by C. infundibulifera on H. rosa-sinenesis has been reported in Japan, Myanmar, Nepal, Guinea, and the United States (1). In Korea, there was one record of this fungus on H. syriacus (1). To our knowledge, this is the first report of C. infundibulifera on H. rosa-sinensis in Korea. This pathogen could be a potential threat to the production of this ornamental shrub over a prolonged period of hot and humid weather. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, Retrieved February 28, 2014. (2) P. M. Kirk. Mycol. Pap. 152:1, 1984. (3) G. Walther et al. Persoonia 30:11, 2013.

scholarly journals First Report of Boeremia Blight Caused by Boeremia exigua var. exigua on Pyrethrum in Australia

Plant Disease ◽  
2011 ◽  
Vol 95 (11) ◽  
pp. 1478-1478 ◽  
Author(s):  
S. J. Jones ◽  
F. S. Hay ◽  
T. C. Harrington ◽  
S. J. Pethybridge
Keyword(s):  
Disease Incidence ◽  
Elongation Factor ◽  
Its Region ◽  
Tween 20 ◽  
Width Ratio ◽  
Malt Extract ◽  
Base Pairs ◽  
Internal Transcribed Spacer Region ◽  
First Report ◽  
Surface Sterilization

Pyrethrum (Tanacetum cinerariifolium) is produced for extraction of insecticidal compounds from the flower achenes. In 2004 and 2006, isolations from necrotic lesions on stems and leaves in three fields in northern Tasmania, Australia yielded four unidentified fungal isolates. Leaf lesions were medium brown and circular (2 to 4 mm in diameter) or irregular in shape (2 to 5 mm long). Stem lesions were irregular, necrotic spots, 5 to 15 mm below the flower peduncle, medium brown, 2 to 4 mm long, and 1 to 2 mm wide. Isolations were conducted on water agar following surface sterilization. Isolates were identified by colony characteristics and the presence of metabolite ‘E’ (1). On oatmeal agar (OA), colonies had irregular margins, were greenish olivaceous-to-olivaceous gray with sparse, white, floccose, aerial mycelia. On malt extract agar (MEA), cultures were variable in color with olivaceous black centers with soft, dense, aerial mycelia. Conidia were hyaline, ellipsoidal to oblong, mainly aseptate, but occasionally 1-septate with dimensions ranging from 2.5 to 7.5 × 1.8 to 3.8 μm (length/width ratio = 1.7 to 2.1). All isolates had moderate reactions to the NaOH test for metabolite ‘E’. DNA was extracted from all four isolates with a DNeasy Plant Mini Kit (QIAGEN Inc., Valencia, CA). For identification, the internal transcribed spacer region (ITS1, 5.8s, and ITS2) and part of the translation elongation factor (TEF) region were amplified and sequenced. Primers ITS1 and ITS4 (2) were used for the ITS region and primers EFCF1 (5′-AGTGCGGTGGTATCGACAAG) and EFCF6 (3′-CATGTCACGGACGGCGAAAC) were used for the TEF. Amplicons were sequenced in both directions and consensus sequences assembled. The ITS sequence was 100% identical to Boeremia exigua var. exigua (GenBank Accession No. GU237715). Base pairs 413 to 1,214 of the TEF sequence from the pyrethrum isolates matched base pairs 1 to 802 (799 of 802 identities) of B. exigua var. exigua (GenBank Accession No. GU349080). All isolates were confirmed as B. exigua var. exigua using morphology and sequencing. Pathogenicity tests were conducted three times in separate glasshouse trials for two of the four isolates. For each isolate, conidial suspensions in water (3 ml/plant) from MEA, adjusted to 5 × 105/ml were applied with Tween 20 (1 drop per 100 ml of water) to 8-week-old pyrethrum plants (five pots per isolate with four plants per pot) using a hand-held spray bottle. Twenty plants were sprayed with water and Tween 20 as nontreated controls. Plants were covered with plastic bags for 48 h after inoculation and examined for symptoms after 15 days at 20°C. Disease incidence (number of symptomatic leaves affected per total number of leaves) of the inoculated plants varied from 7.5 to 9.4%. Noninoculated plants did not develop symptoms. Isolations resulted in cultures morphologically identical on MEA and OA to those inoculated. To our knowledge, this is the first report of B. exigua var. exigua causing disease in pyrethrum. Cultures were deposited in the New South Wales Department of Agriculture collection (DAR79101 to 79104) and TEF and ITS sequences for DAR79101 in GenBank (Accession Nos. JF925328 and JF925329, respectively). Boeremia blight is likely to contribute to the fungal disease complex causing reductions in green leaf area in Australian pyrethrum production. References: (1) M. M. Aveskamp et al. Stud. Mycol. 65:1, 2010. (2) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

scholarly journals First Report of Neofusicoccum australe and N. luteum Associated with Canker and Dieback of Quercus robur in Portugal

Plant Disease ◽  
2013 ◽  
Vol 97 (4) ◽  
pp. 560-560 ◽  
Author(s):  
C. Barradas ◽  
A. Correia ◽  
A. Alves
Keyword(s):  
Quercus Robur ◽  
Elongation Factor ◽  
Aerial Mycelium ◽  
Its Region ◽  
Deciduous Tree ◽  
Pedunculate Oak ◽  
Single Spore ◽  
Canker Disease ◽  
First Report ◽  
Wide Range

Quercus robur L., commonly known as “pedunculate oak,” is a long-lived deciduous tree native to most of Europe. It is of great ecological and forestry importance. It is also commonly cultivated as an ornamental tree in parks and gardens. Since 2009 and most likely related to increased drought periods, diseased ornamental trees have been observed in the campus of the University of Aveiro, Portugal. More than 50% of the trees are already damaged by the disease. The symptoms included twig and branch dieback and sunken necrotic bark lesions that could progress to the trunk, resulting in the death of large sections of the tree. Ascomata and conidiomata typical of Botryosphaeriaceae were observed on branches of symptomatic trees. Ascospores were hyaline, aseptate, ovoid to fusoid, and conidia were hyaline, aseptate, smooth, thin-walled, and fusiform with base truncate. Single spore isolates were obtained from samples. In culture, single ascospore isolates produced conidia similar to the ones found on the host. Diseased branch tissues were surface sterilized with 5% NaOCl, plated on potato dextrose agar (PDA), and incubated at 25°C. Fungal isolates recovered produced white aerial mycelium that darkened with age becoming grey to dark grey and conidia that were similar in all aspects to the ones produced by single spore isolates. All isolates produced on PDA a yellow pigment that diffused into the agar and disappeared after 6 to 7 days. Morphological and cultural aspects of the isolates were similar to the species Neofusicoccum luteum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips and N. australe (Slippers, Crous & M.J. Wingf.) Crous, Slippers & A.J.L. Phillips (2). Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR fingerprinting divided the isolates into two groups (1). Two isolates (CAA352 and CAA392), one from each group, were selected for further identification by sequencing of the internal transcribed spacer (ITS) region of the rDNA and part of the elongation factor 1-alpha gene (EF1-α) (Accession No. ITS: JX975212 JX975213; EF1-α: JX975210 JX975211). BLAST analysis showed that ITS and EF1-α sequences from group 1 and 2 had 99 to 100% similarity to reference cultures (including ex-type) of N. luteum and N. australe, respectively. To confirm pathogenicity and fulfill Koch's postulates, six 2-year-old seedlings of Q. robur were artificially infected with isolates CAA352 and CAA392 and kept at approximately 20 to 25°C. A shallow wound was done with a scalpel on the basal part of the stem of each seedling, a bark portion was removed aseptically and a PDA disc (0.5 cm) of an actively growing culture was placed on the wound. Control seedlings received sterile PDA discs. The inoculation site was wrapped in Parafilm to prevent desiccation. Within 8 weeks, infected seedlings developed canker lesions associated with vascular necrosis around the inoculation point. A third of the seedlings died and developed abundant pycnidia on the stem. Control seedlings remained symptomless. Both pathogens were successfully reisolated from the infected tissue. N. luteum and N. australe are increasingly reported as causing diseases to a wide range of woody hosts of economic and forestry importance (3). To our knowledge, this is the first report of both species causing dieback and canker disease on Q. robur. References: (1) A. Alves et al. Res. Microbiol. 158:112, 2007. (2) P. W. Crous et al. Stud. Mycol. 55:235, 2006. (3) B. Slippers et al. Fungal Biol. Rev. 21:90, 2007.

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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