scholarly journals Morphological and molecular identification of Fusarium spp. isolated from maize kernels in Java and Lombok, Indonesia

2020 ◽  
Vol 21 (6) ◽  
Author(s):  
Ani Widiastuti ◽  
Monica Lucky Karlina ◽  
Kurnia Ritma Dhanti ◽  
Yufita Dwi Chinta ◽  
Tri Joko ◽  
...  
Keyword(s):  
Molecular Identification ◽  
Fusarium Verticillioides ◽  
Animal Health ◽  
Morphological Characteristics ◽  
Morphological Identification ◽  
Fusarium Spp ◽  
Single Spore ◽  
Central Java ◽  
Species Specific ◽  
Maize Kernels

Abstract. Widiastuti A, Karlina ML, Dhanti KR, Chinta YD, Joko T, Suryanti, Wibowo A. 2020. Morphological and molecular identification of Fusarium spp. isolated from maize kernels in Java and Lombok, Indonesia. Biodiversitas 21: 2741-2750. Fungal contamination of maize is a serious problem in Indonesia. Fusarium spp. infect maize in the field will be continuing to contaminate in the post-harvest period even though disease symptoms are not always emerged. Some Fusarium spp. produced mycotoxins which are harmful to human and animal health. Aims of this research were to reveal the presence of Fusarium spp. from both symptomatic and unsymptomatic maize, and to identify them based on morphological characteristics and molecular analysis. Samples of maize were collected from maize cultivation areas in East Java (EJ), Central Java (CJ), West Java (WJ), Yogyakarta Special Province (DIY), and Lombok, West Nusa Tenggara. Fusarium spp. were isolated in a single spore method and cultured in potato dextrose agar (PDA) medium for morphological identification of macro-and microconidia. Molecular identification was conducted by PCR assay using species-specific primers. Furthermore, unidentified species were analyzed by DNA sequence. This research found four species of mycotoxigenic Fusarium isolated from maize-based on molecular identification, which were Fusarium verticillioides (15 isolates), F. proliferatum (6 isolates), F. graminearum (1 isolate) and F. asiaticum (1 isolate). This research showed a novel report of F. asiaticum infection on maize kernel in Indonesia.

scholarly journals Survey for toxigenic Fusarium species on maize kernels in China

2020 ◽  
Vol 13 (2) ◽  
pp. 213-224 ◽  
Author(s):  
P.W. Qin ◽  
J. Xu ◽  
Y. Jiang ◽  
L. Hu ◽  
T. van der Lee ◽  
...  
Keyword(s):  
Seed Quality ◽  
Fusarium Species ◽  
Fusarium Verticillioides ◽  
Morphological Characteristics ◽  
Ear Rot ◽  
Maize Production ◽  
Fusarium Asiaticum ◽  
Maize Varieties ◽  
Species Specific ◽  
Maize Kernels

Maize is currently the most important crop in China. A major concern in maize production is maize ear rot caused by Fusarium spp., which results in yield losses, reduction of seed quality and the accumulation of mycotoxins in the harvested grains. To identify the importance of the different Fusarium species in maize infection, we performed a comprehensive survey on 9,000 asymptomatic and randomly collected maize kernels. Seeds were collected from 12 different provinces covering all major maize growing areas in China and included five maize varieties. In total 1,022 Fusarium isolates were retrieved that were identified based on morphological characteristics, by species specific diagnostic PCRs and by EF1-α gene sequencing. Eight different species were identified: Fusarium verticillioides (75.34%), Fusarium graminearum (8.32%), Fusarium proliferatum (7.14%), Fusarium subglutinans (4.11%), Fusarium meridionale (1.57%), Fusarium oxysporum (1.37%), Fusarium semitectum (1.17%), and Fusarium asiaticum (0.98%). The distribution of Fusarium species was found to be different in different regions with the largest diversity observed in Hubei province, where all eight Fusarium species were isolated. Genetic chemotyping within the F. graminearum species complex indicated that all of the 85 F. graminearum isolates showed the 15-acetyldeoxynivalenol chemotype, whereas all F. asiaticum (n=10) and F. meridionale (n=16) isolates had the nivalenol chemotype even when isolated from the same maize field. To our knowledge this is the largest collection of Fusarium isolates from maize and further exploitations of this collection are discussed.

scholarly journals First Report of Cercospora apii Leaf Spot on Capsicum chinense in Brazil

Plant Disease ◽  
2011 ◽  
Vol 95 (9) ◽  
pp. 1194-1194 ◽  
Author(s):  
A. Nicoli ◽  
L. Zambolim ◽  
E. G. C. Nasu ◽  
D. B. Pinho ◽  
O. L. Pereira ◽  
...  
Keyword(s):  
Leaf Spot ◽  
Morphological Characteristics ◽  
Minas Gerais ◽  
Hot Pepper ◽  
Capsicum Chinense ◽  
Single Spore ◽  
Cornell University ◽  
Hot Peppers ◽  
C 30 ◽  
Species Specific

In Brazil, Capsicum chinense Jacq. is the predominant species of commercial hot peppers because of its popular citrus-like aroma and adaptability to different soils and climates (4). In June 2010, 30 samples of C. chinense with severe leaf spot were collected from a field in the city of Viçosa, state of Minas Gerais, Brazil. Symptoms were observed on leaves, calyxes, fruits, and stems on most of the plants found in the area. On leaves, symptoms included amphigenous lesions that were initially circular to ellipsoid, 1 to 5 mm in diameter, whitish to tan in the center, and surrounded by a dark brown or reddish purple border. Lesions coalesce and turned necrotic with age. A fungus isolated from the lesions matched well with the description of Cercospora apii Fresen. It formed erumpent stromata that were dark brown and spherical to irregular; fascicule conidiophores were clear brown or pale, straight or curved, unbranched, geniculate, 22.5 to 80 × 5 to 7.5 μm, 0 to 3 septate, subtruncate apex; and conidia were solitary, hyaline to subhyaline, filiform, base truncate, tip acute, straight to curved, 12.5 to 140 × 3.5 to 5 μm, and 0 to 11 septate (1,2). A sample was deposited in the herbarium of the Universidade Federal de Viçosa, Minas Gerais, Brazil (VIC 31415). Identity was confirmed by amplifying part of the calmodulin gene with species-specific primers CercoCal-apii and CercoCal-R (3) of fungal DNA from a single-spore culture. In amplification reaction, initial denaturation step was done at 94°C for 5 min, followed by 40 cycles of denaturation at 94°C (30 s), annealing at 56°C (30 s), and elongation at 72°C (30 s). Primers CercoCal-apii and CercoCal-R amplified a single DNA product of 176 bp, and coupled with the morphological characteristics, confirmed the identity of the fungus as Cercospora apii. To check pathogenicity, a 6-mm-diameter plug of the isolate was removed from the expanding edge of a 21-day-old culture grown on potato dextrose agar (PDA) and placed in contact with the adaxial face of the leaves of 8-week-old C. chinense grown in 2-liter plastic pots with soil substrate. Six plants, one per pot, were inoculated with the isolate and six plants were inoculated with the fungus-free PDA plug. Inoculated plants were maintained in a moist chamber for 24 h and then subsequently kept in a greenhouse at 26°C. Leaf spot was observed in all inoculated plants 15 days after inoculation and symptoms were similar to those expressed in the field. The fungus was reisolated from the inoculated plants and matched well with the description of Cercospora apii. All fungus-free PDA inoculated plants remained healthy. Cercospora apii comprises a complex of 281 morphologically indistinguishable species that can infect an extremely wide host range (2). To our knowledge, this pathogen has the potential to cause significant damage to the hot pepper industry of Brazil. References: (1) C. Chupp. A Monograph of the Fungus Cercospora. Cornell University Press, Ithaca, NY, 1954. (2) P. W. Crous and U. Braun. CBS Biodivers. Ser. 1:1, 2003. (3) M. Groenewald et al. Phytopathology 95:951, 2005. (4) S. D. Lannes et al. Sci. Hortic. 112:266, 2007.

scholarly journals Molecular identification of Auxis spp. larvae (Pisces: Scombridae) from the Gulf of California: Solving morphological identification limits

2018 ◽  
Vol 53 (2) ◽  
pp. 157 ◽  
Author(s):  
María J. Ochoa-Muñoz ◽  
Noé Díaz-Viloria ◽  
Laura Sánchez-Velasco ◽  
Sylvia P. A. Jiménez-Rosenberg ◽  
Ricardo Pérez-Enríquez
Keyword(s):  
Molecular Identification ◽  
Gulf Of California ◽  
Fish Larvae ◽  
Morphological Characteristics ◽  
Species Level ◽  
Morphological Identification ◽  
Larval Stages ◽  
Marine Fish Larvae ◽  
Coi Sequences ◽  
First Time

The larvae of the Auxis genus are abundant in the Gulf of California during summer; however, their identification to the species level by morphological methods is a challenge. The goal of this study was to identify A. thazard and A. rochei larvae for first time, through molecular markers using COI sequences of mtDNA, and look for distinctive morphological characteristics between species, mainly in pigmentation patterns. Larvae were obtained by zooplankton tows in 3 oceanographic cruises in the southern Gulf of California and adjacent waters. The presence of A. thazard and A. rochei larvae was genetically confirmed. The sequences of 7 larvae showed genetic divergences lower than 1% when were compared to sequences of A. thazard adults, while 15 larvae showed genetic divergences lower than 2% when where compared to sequences of A. rochei adults. Genetic divergences between both Auxis species were higher than 2%. These results suggest the spawning of both species in the Gulf of California. On the other hand, pigmentation patterns and morphometric characteristics, in all larval stages, did not permit the secure differentiation between species. Thus, the use of molecular identification by COI is recommended to identify Auxis larvae to the species level, as well as in other marine fish larvae collected in other regions of the world, that have identification troubles.

PCR Detection Assay of Fumonisin-Producing Fusarium verticillioides Strains

2004 ◽  
Vol 67 (6) ◽  
pp. 1278-1283 ◽  
Author(s):  
BELÉN PATIÑO ◽  
SALVADOR MIRETE ◽  
M. TERESA GONZÁLEZ-JAÉN ◽  
GIUSEPPINA MULÉ ◽  
M. TERESA RODRÍGUEZ ◽  
...  
Keyword(s):  
Fusarium Species ◽  
Fusarium Verticillioides ◽  
Animal Health ◽  
Intergenic Spacer ◽  
Fungal Species ◽  
Intergenic Spacer Region ◽  
Detection And Identification ◽  
Geographical Regions ◽  
Species Specific ◽  
A Minor

Fusarium verticillioides is considered to be the main source of fumonisins, a group of toxins that contaminate commodities and result in chronic and acute diseases affecting humans and animals. The detection and control of this species is crucial to prevent fumonisins from entering the food chain. The objective of the present research was to develop a specific, sensitive, and robust PCR assay to detect F. verticillioides strains using two pairs of specific primers for F. verticillioides, which have been designed on the basis of the intergenic spacer region of the rDNA units. The first pair of primers was F. verticillioides species specific, whereas the second pair of primers detected fumonisin-producing F. verticillioides strains. This second pair of primers allowed for the discrimination between the major group of F. verticillioides strains, fumonisin-producing strains that are mainly associated with crops, and a minor group of strains, non–fumonisin-producing strains that are associated with bananas. Fifty-four strains of F. verticillioides from different geographical regions and hosts were tested using both sets of primers. Sixteen additional Fusarium species were examined. The specificity of the primer sequences provides the basis for a simple, rapid, accurate, and sensitive detection and identification method of this fungal species that represents a risk for human and animal health.

scholarly journals First Report of Cercospora Leaf Spot Caused by Cercospora apii (= C. molucellae) on Bells-of-Ireland (Molucella laevis) in Mexico

Plant Disease ◽  
2009 ◽  
Vol 93 (2) ◽  
pp. 197-197 ◽  
Author(s):  
V. Ayala-Escobar ◽  
U. Braun ◽  
C. Nava-Diaz
Keyword(s):  
Uv Light ◽  
Morphological Characteristics ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Cercospora Leaf Spot ◽  
Single Spore ◽  
First Report ◽  
Cornell University ◽  
Specific Fragment ◽  
Species Specific

In late 2007, a new disease was found in commercial cutflower fields of bells-of-Ireland (Molucella laevis L.) in Texcoco, Mexico. Four plantings surveyed during this time had 100% incidence. A few spots on cutflowers make them unmarketable. Symptoms consisted of gray-green spots on leaves, calyxes, and stems, which turned brown with age. Spots were initially circular to oval, delimited by major leaf veins, and were visible on both adaxial and abaxial sides of the leaves. A Cercospora species was consistently associated with the spots. The fungus was isolated on V8 agar medium. Three single-spore cultures were obtained from isolation cultures. Cultures were incubated at 24°C under near-UV light for 7 days. Pathogenicity was confirmed by spraying a conidial suspension (1 × 104 condia/ml) on leaves of 16 potted M. laevis plants, incubating the plants in a dew chamber for 48 h, and maintaining them in a greenhouse (20 to 24°C). Identical symptoms to those observed in the field appeared on all inoculated plants after 2 weeks. No symptoms developed on control plants treated with autoclaved distilled water. The pathogenicity test was repeated twice with similar results. The fungus produced erumpent stromata, which were dark brown, spherical to irregular, 10 to 26 μm diameter, and giving rise to fascicles of five to nine divergent conidiophores, which were clear brown, paler near the subtruncate apex, straight to curved, not branched, rarely geniculate with two to four septa, and 57 × 3.4 μm. The conidia were formed singly, hyaline, acicular, base truncate, tip acute, straight to curved with 11 to 19 septa, and 172 × 3.5 μm. Fungal DNA from single-spore cultures was obtained with a commercial extraction kit (Qiagen, Hilden, Germany), amplified with ITS5 and ITS4 primers, and sequenced. The sequence, deposited at the National Center for Biotechnology Information Database (GenBank Accession No. EU564808), aligned almost perfectly (99% identity) to the bells-of-Ireland isolates from California (GenBank Accession Nos. AY156918 and AY156919) and New Zealand (Accession No. DQ233321). A 176-bp species-specific fragment was amplified with CercoCal-apii primers but not with CercoCal-beta or CercoCal-sp primers. These results, coupled with the morphological characteristics (1) and pathogenicity test, confirm the identity of the fungus as Cercospora apii sensu lato (including C. molucellae) (2,3,4). Although C. apii sensu lato has been reported on other hosts in Mexico (1,2), to our knowledge, this is the first report of this disease on M. laevis plants in this country. References: (1) C. Chupp. A Monograph of the Fungus Cercospora. Cornell University Press, Ithaca, NY, 1954. (2) P. W. Crous and U. Braun. CBS Biodiversity Series 1:1, 2003. (3) M. Groenewald et al. Phytopathology 95:951, 2005. (4) S. T Koike et al. Plant Dis. 87:203, 2003.

scholarly journals Amylopectin Induces Fumonisin B1 Production by Fusarium verticillioides During Colonization of Maize Kernels

2005 ◽  
Vol 18 (12) ◽  
pp. 1333-1339 ◽  
Author(s):  
B. H. Bluhm ◽  
C. P. Woloshuk
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Fusarium Verticillioides ◽  
Animal Health ◽  
Fumonisin B1 ◽  
Polymerase Chain Reaction Analysis ◽  
Defined Medium ◽  
Wild Type ◽  
Fungal Genes ◽  
High Amylose ◽  
Maize Kernels

Fusarium verticillioides, a fungal pathogen of maize, produces fumonisin mycotoxins that adversely affect human and animal health. Basic questions remain unanswered regarding the interactions between the host plant and the fungus that lead to the accumulation of fumonisins in maize kernels. In this study, we evaluated the role of kernel endosperm composition in regulating fumonisin B1 (FB1) biosynthesis. We found that kernels lacking starch due to physiological immaturity did not accumulate FB1. Quantitative polymerase chain reaction analysis indicated that kernel development also affected the expression of fungal genes involved in FB1 biosynthesis, starch metabolism, and nitrogen regulation. A mutant strain of F. verticillioides with a disrupted α-amylase gene was impaired in its ability to produce FB1 on starchy kernels, and both the wild-type and mutant strains produced significantly less FB1 on a high-amylose kernel mutant of maize. When grown on a defined medium with amylose as the sole carbon source, the wild-type strain produced only trace amounts of FB1, but it produced large amounts of FB1 when grown on amylopectin or dextrin, a product of amylopectin hydrolysis. We conclude that amylopectin induces FB1 production in F. verticillioides. This study provides new insight regarding the interaction between the fungus and maize kernel during pathogenesis and highlights important areas that need further study.

scholarly journals First Report of Stem Rot of Papaya Caused by Fusarium solani Species Complex in Brazil

Plant Disease ◽  
2013 ◽  
Vol 97 (1) ◽  
pp. 140-140 ◽  
Author(s):  
K. C. Correia ◽  
B. O. Souza ◽  
M. P. S. Câmara ◽  
S. J. Michereff
Keyword(s):  
Dna Sequences ◽  
Fusarium Solani ◽  
Species Complex ◽  
Morphological Characteristics ◽  
Molecular Techniques ◽  
Northeastern Brazil ◽  
Stem Rot ◽  
Morphological Identification ◽  
Single Spore ◽  
First Report

In October 2010, 2-year-old papaya (cv. Hawaii) trees with high incidence of stem rot were observed during a survey conducted in Rio Grande do Norte state, northeastern Brazil. Stems showing reddish brown-to-dark brown symptoms were collected and small pieces (4 to 5 mm) of necrotic tissues were surface sterilized for 1 min in 1.5% NaOCl, washed twice with sterile distilled water, and plated onto potato dextrose agar (PDA) amended with 0.5 g liter–1 streptomycin sulfate. Plates were incubated at 25°C with a 12-h photopheriod for 4 days. Pure cultures with white, fluffy aerial mycelia were obtained by subculturing hyphal tips onto PDA. Identification was made using morphological characteristics and DNA based molecular techniques. Colonies grown on PDA and Spezieller Nährstoffarmer agar (SNA) for 10 days at 25°C with a 12-h photoperiod were used for morphological identification (3). The fungus produced cream sporodochia and two types of spores: microconidia were thin-walled, hyaline, ovoid, one-celled, and 6.8 to 14.6 × 2.3 to 4.2 μm; macroconidia were thick walled, hyaline, slightly curved, 3- to 5-celled, and 25.8 to 53.1 × 3.9 to 5.7 μm. Fifty spores of each type were measured. Rounded, thick-walled chlamydospores were produced, with two to four arranged together. On the basis of morphological characteristics (1), three fungal isolates (CMM-3825, CMM-3826, and CMM-3827) were identified as Fusarium solani (Mart.) Sacc. and were deposited in the Culture Collection of Phytopathogenic Fungi of the Universidade Federal Rural de Pernambuco (Recife, Brazil). Single-spore isolates were obtained and genomic DNA of the isolates was extracted and a portion of the translation elongation factor 1-alpha (EF1-α) gene of the isolates was amplified and sequenced (2). When compared with sequences available in the GenBank and Fusarium-ID databases, DNA sequences of the three isolates shared 99 to 100% sequence identity with F. solani species complex (GenBank Accession Nos. JF740784.1, DQ247523.1, and DQ247017.1). Representative sequences of the isolates were deposited in GenBank (Accession Nos. JQ808499, JQ808500, and JQ808501). Pathogenicity tests were conducted with four isolates on 3-month-old papaya (cv. Hawaii) seedlings. Mycelial plugs taken from the margin of actively growing colonies (PDA) of each isolate were applied in shallow wounds (0.4 cm in diameter) on the stem (center) of each plant. Inoculation wounds were wrapped with Parafilm. Control seedlings received sterile PDA plugs. Inoculated and control seedlings (10 each) were kept in a greenhouse at 25 to 30°C. After 2 weeks, all inoculated seedlings showed reddish brown necrotic lesions in the stems. No symptoms were observed in the control plants. The pathogen was successfully reisolated from symptomatic plants to fulfill Koch's postulates. To our knowledge, this is the first report of F. solani species complex causing papaya stem rot in Brazil. Papaya is an important fruit crop in the northeastern Brazil and the occurrence of this disease needs to be taken into account in papaya production. References: (1) C. Booth. Fusarium Laboratory Guide to the Identification of the Major Species. CMI, Kew, England, 1977. (2) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006.

Influence of nitrogen rates and Fusarium verticillioides infection on Fusarium spp. and fumonisin contamination of maize kernels

2021 ◽  
Vol 144 ◽  
pp. 105601
Author(s):  
Vesna Krnjaja ◽  
Violeta Mandić ◽  
Zorica Bijelić ◽  
Slavica Stanković ◽  
Ana Obradović ◽  
...  
Keyword(s):  
Fusarium Verticillioides ◽  
Fusarium Spp ◽  
Nitrogen Rates ◽  
Maize Kernels

scholarly journals Antifungal sensitivity profile of Fusarium spp. resulting keratitits

2017 ◽  
Vol 1 (1) ◽  
Keyword(s):  
Amphotericin B ◽  
Antifungal Agents ◽  
Antifungal Susceptibility ◽  
Morphological Characteristics ◽  
Fungal Keratitis ◽  
Morphological Identification ◽  
Fusarium Spp ◽  
Sensitivity Testing ◽  
Resistant Strains ◽  
Emergence Of Resistance

Fungal keratitis is an important cause of visual impairment and blindness. Genus Fusarium is a leading cause for fungal keratitis and it has higher degree of resistance to antifungal agents. Our objectives were to identify Fusarium spp. isolated from corneal specimens (received at Dept. of Mycology - MRI from 2013-2016) up to species level and to determine antifungal susceptibility pattern of them. All Fusarium isolates (51) obtained from specimens of patients with keratitis were included in the study. Speciation was done using morphological characteristics of fungi. Antifungal sensitivity testing was done according to CLSI M 51- A guideline, against amphotericin B (10 µg), itraconazole (10 µg). and voriconazole (1 µg). Majority of the isolates were F. solani complex (n=24). Three isolates were difficult to speciate morphologically. Significant number of Fusarium isolates had inhibitory zone diameters (IZD) less than tentative zone diameter epidemiological cut off values (TZD ECVs) for both itraconazole and amphotericin B, indicating emergence of resistant strains against these drugs. Forty five isolates (97.82%) had IZD more than corresponding TZD ECV for voriconazole. All F. solani complexes had IZD less than TZD ECVs for itraconazole. Morphological identification cannot be used as the only method for speciation of Fusarium isolates. Antifungal sensitivity testing should be done for Fusarium isolates from keratitis patients as emergence of resistance strains is not uncommon against commonly used antifungal agents.

scholarly journals Validation of an Effective Protocol for Culicoides Latreille (Diptera: Ceratopogonidae) Detection Using eDNA Metabarcoding

Insects ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 401
Author(s):  
Yoamel Milián-García ◽  
Lauren A. A. Janke ◽  
Robert G. Young ◽  
Aruna Ambagala ◽  
Robert H. Hanner
Keyword(s):  
Molecular Identification ◽  
Molecular Detection ◽  
Morphological Characteristics ◽  
Species Level ◽  
Morphological Identification ◽  
Culicoides Species ◽  
Nacl Solution ◽  
Identification Method

eDNA metabarcoding is an effective molecular-based identification method for the biosurveillance of flighted insects. An eDNA surveillance approach maintains specimens for secondary morphological identification useful for regulatory applications. This study identified Culicoides species using eDNA metabarcoding and compared these results to morphological identifications of trapped specimens. Insects were collected using ultraviolet (UV) lighted fan traps containing a saturated salt (NaCl) solution from two locations in Guelph, Ontario, Canada. There were forty-two Culicoides specimens collected in total. Molecular identification detected four species, C. biguttatus, C. stellifer, C. obsoletus, and C. mulrennani. Using morphological identification, two out of these four taxonomic ranks were confirmed at the species level (C. biguttatus and C. stellifer) and one was confirmed at the subgenus level (Avaritia [C. obsoletus]). No molecular detection of Culicoides species occurred in traps with an abundance of less than three individuals per taxon. The inconsistency in identifying Culicoides specimens to the species level punctuates the need for curated DNA reference libraries for Culicoides. In conclusion, the saturated salt (NaCl) solution preserved the Culicoides’ morphological characteristics and the eDNA.

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