scholarly journals Development of PCR-Based Race-Specific Markers for Differentiation of Indian Fusarium oxysporum f. sp. cubense, the Causal Agent of Fusarium Wilt in Banana

2022 ◽  
Vol 8 (1) ◽  
pp. 53
Author(s):  
Raman Thangavelu ◽  
Esack Edwinraj ◽  
Muthukathan Gopi ◽  
Periyasamy Pushpakanth ◽  
Kotteswaran Sharmila ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Fusarium Oxysporum ◽  
Fusarium Wilt ◽  
Fusarium Species ◽  
In Silico Analysis ◽  
Specific Marker ◽  
In Planta ◽  
Variable Regions ◽  
Vegetative Compatibility Groups ◽  
Highly Virulent

Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc), is the most lethal soil-borne fungal pathogen infecting bananas. Foc race 1 (R1) and 4 (R4) are the two most predominant races affecting the economically important Cavendish group of bananas in India. A total of seven vegetative compatibility groups (VCGs) from three pathogenic races were isolated during our field survey and were found to be highly virulent towards cv. Grande Naine. According to comparative genome analyses, these Indian Foc VCGs were diverse in genomic organization and effector gene profiles. As a result, false-positive results were obtained with currently available molecular markers. In this context, the study has been initiated to develop PCR-based molecular markers for the unambiguous identification of Indian Foc R1 and R4 isolates. Whole-genome sequences of Foc R1 (GCA_011316005.3), Foc TR4 (GCA_014282265.3), and Foc STR4 (GCA_016802205.1), as well as the reference genomes of Foc (ASM799451v1) and F. oxysporum f. sp. lycopersici (Fol; ASM14995v2), were aligned to identify unique variable regions among the Foc races. Using putative chromosome and predicted gene comparison, race-specific unique Foc virulence genes were identified. The putative lineage-specific identified genes encoding products secreted in xylem (SIX) that may be necessary for disease development in the banana. An in silico analysis was performed and primers were designed from a region where sequences were dissimilar with other races to develop a specific marker for Foc R1, R4, TR4, and STR4. These race-specific markers allowed target amplification in the characterized highly virulent Foc isolates, and did not show any cross-amplification to any other Foc races, VCGs or banana pathogens, Fusarium species, and non-pathogenic Fusarium oxysporum isolates. The study demonstrated that the molecular markers developed for all the three Foc races of India could detect the pathogen in planta and up to 0.025 pg µL−1 DNA levels. Thus, the markers developed in this study are novel and could potentially be useful for the accurate diagnosis and detection of the Indian Foc races which are important for the effective management of the disease.

scholarly journals Current Classification and Diversity of Fusarium Species Complex, the Causal Pathogen of Fusarium Wilt Disease of Banana in Malaysia

Agronomy ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 1955
Author(s):  
Anysia Hedy Ujat ◽  
Ganesan Vadamalai ◽  
Yukako Hattori ◽  
Chiharu Nakashima ◽  
Clement Kiing Fook Wong ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Fusarium Wilt ◽  
Species Complex ◽  
Phylogenetic Trees ◽  
Fusarium Species ◽  
Morphological Characteristics ◽  
Pathogenicity Test ◽  
Fusarium Spp ◽  
Specific Primers ◽  
Fusarium Oxysporum Species Complex

The re-emergence of the Fusarium wilt caused by Fusarium odoratissimum (F. odoratissimum) causes global banana production loss. Thirty-eight isolates of Fusarium species (Fusarium spp.) were examined for morphological characteristics on different media, showing the typical Fusarium spp. The phylogenetic trees of Fusarium isolates were generated using the sequences of histone gene (H3) and translation elongation factor gene (TEF-1α). Specific primers were used to confirm the presence of F. odoratissimum. The phylogenetic trees showed the rich diversity of the genus Fusarium related to Fusarium wilt, which consists of F. odoratissimum, Fusarium grosmichelii, Fusarium sacchari, and an unknown species of the Fusarium oxysporum species complex. By using Foc-TR4 specific primers, 27 isolates were confirmed as F. odoratissimum. A pathogenicity test was conducted for 30 days on five different local cultivars including, Musa acuminata (AAA, AA) and Musa paradisiaca (AAB, ABB). Although foliar symptoms showed different severity of those disease progression, vascular symptoms of the inoculated plantlet showed that infection was uniformly severe. Therefore, it can be concluded that the Fusarium oxysporum species complex related to Fusarium wilt of banana in Malaysia is rich in diversity, and F. odoratissimum has pathogenicity to local banana cultivars in Malaysia regardless of the genotype of the banana plants.

scholarly journals The development of a multiplex PCR assay for the detection of Fusarium oxysporum f. sp. cubense lineage VI strains in East and Central Africa

2020 ◽  
Vol 158 (2) ◽  
pp. 495-509 ◽  
Author(s):  
P. Ndayihanzamaso ◽  
P. Karangwa ◽  
D. Mostert ◽  
G. Mahuku ◽  
G. Blomme ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Multiplex Pcr ◽  
Fusarium Wilt ◽  
Central Africa ◽  
East African ◽  
Pcr Assay ◽  
Vegetative Compatibility Groups ◽  
Worldwide Population ◽  
Multiplex Pcr Assay ◽  
Disease Free

Abstract Banana is a staple food and cash crop grown in East and Central Africa (ECA). The main banana varieties grown in ECA are the East African highland bananas (EAHB), although dessert/beer bananas such as Sukari Ndizi, Kayinja (Pisang Awak) and Gros Michel are also produced due to their high value at local markets. The Fusarium wilt fungus Fusarium oxysporum f. sp. cubense (Foc) causes disease of susceptible dessert/beer bananas, which significantly reduces yields. Banana Fusarium wilt is managed by excluding the pathogen from disease-free areas and by planting disease-resistant varieties in infested fields. Six phylogenetically closely-related vegetative compatibility groups (VCGs) of Foc, VCGs 0124, 0125, 0128, 01212, 01220 and 01222 are present in ECA, which all group together in Foc Lineage VI. Rapid and accurate detection of Foc Lineage VI strains is thus important to prevent its spread to disease-free areas. In this study, molecular markers specific to Foc Lineage VI were therefore developed. Primer sets were then combined in a multiplex PCR assay, and validated on a worldwide population of 623 known Foc isolates, other formae speciales and non-pathogenic Fusarium oxysporum isolates. The Foc Lineage VI multiplex PCR was used to identify Foc isolates collected in banana fields at five locations in Uganda and Tanzania. Foc Lineage VI DNA was detected at a concentration as low as 0.1 ng/μl, both in the absence and presence of banana DNA, and can therefore be used as an accurate diagnostic tool for Foc Lineage VI strains.

scholarly journals In Planta and Soil Quantification of Fusarium oxysporum f. sp. ciceris and Evaluation of Fusarium Wilt Resistance in Chickpea with a Newly Developed Quantitative Polymerase Chain Reaction Assay

2011 ◽  
Vol 101 (2) ◽  
pp. 250-262 ◽  
Author(s):  
Daniel Jiménez-Fernández ◽  
Miguel Montes-Borrego ◽  
Rafael M. Jiménez-Díaz ◽  
Juan A. Navas-Cortés ◽  
Blanca B. Landa
Keyword(s):  
Polymerase Chain Reaction ◽  
Fusarium Oxysporum ◽  
Fusarium Wilt ◽  
Plant Root ◽  
Quantitative Polymerase Chain Reaction ◽  
Infested Soil ◽  
In Planta ◽  
Field Plot ◽  
Chain Reaction ◽  
Polymerase Chain

Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris can be managed by risk assessment and use of resistant cultivars. A reliable method for the detection and quantification of F. oxysporum f. sp. ciceris in soil and chickpea tissues would contribute much to implementation of those disease management strategies. In this study, we developed a real-time quantitative polymerase chain reaction (q-PCR) protocol that allows quantifying F. oxysporum f. sp. ciceris DNA down to 1 pg in soil, as well as in the plant root and stem. Use of the q-PCR protocol allowed quantifying as low as 45 colony forming units of F. oxysporum f. sp. ciceris per gram of dry soil from a field plot infested with several races of the pathogen. Moreover, the q-PCR protocol clearly differentiated susceptible from resistant chickpea reactions to the pathogen at 15 days after sowing in artificially infested soil, as well as the degree of virulence between two F. oxysporum f. sp. ciceris races. Also, the protocol detected early asymptomatic root infections and distinguished significant differences in the level of resistance of 12 chickpea cultivars that grew in that same field plot infested with several races of the pathogen. Use of this protocol for fast, reliable, and cost-effective quantification of F. oxysporum f. sp. ciceris in asymptomatic chickpea tissues at early stages of the infection process can be of great value for chickpea breeders and for epidemiological studies in growth chambers, greenhouses and field-scale plots.

Vegetative compatibility groups within Fusarium oxysporum f.sp. lini from Linum usitatissimum (flax) wilt nurseries in western Canada

2001 ◽  
Vol 79 (7) ◽  
pp. 836-843 ◽  
Author(s):  
S I Mpofu ◽  
K Y Rashid
Keyword(s):  
Population Structure ◽  
Fusarium Oxysporum ◽  
Fusarium Wilt ◽  
Linum Usitatissimum ◽  
Vegetative Compatibility ◽  
Western Canada ◽  
Disease Development ◽  
Vegetative Compatibility Groups ◽  
Linum Usitatissimum L ◽  
Compatibility Analysis

Following the discovery of substantial differences in the development and severity of Fusarium wilt in Linum usitatissimum L. (flax) wilt nurseries in western Canada, a study of the population structure of Fusarium oxysporum f.sp. lini (Bolley) Snyd. & Hans using vegetative compatibility analysis was initiated. Vegetative compatibility was determined using nitrate non-utilizing mutants. From a total of 105 isolates, 74 were assigned to 12 vegetative compatibility groups (VCG 0440-04411), 22 were not compatible with any other isolates and 9 did not produce mutants. The populations of F. oxysporum f.sp. lini in Fusarium wilt nurseries in western Canada were significantly different. There was a predominant VCG in each of the nurseries, which was either nonexistent or not common in other nurseries, VCG 0440 in Indian Head, 0441 in Treherne, 0442 in Morden-80, 0443 in Saskatoon, and 0444 in Morden-60. There were only three overlapping VCGs; VCG 0440 was common to Morden-60 and Indian Head and VCGs 0442 and 0444 were common to Morden-60 and Morden-80. The differences in disease development and severity observed previously may be due to the differences in the population structure of the pathogen. These findings have significant implications for the approaches to breeding for Fusarium wilt resistance in flax.Key words: Fusarium oxysporum f.sp. lini, flax, Fusarium wilt, genetic diversity, vegetative compatibility.

Cell wall degrading enzymes in fusarium wilt of chickpea: correlation between pectinase and xylanase activities and disease development in plants infected with two pathogenic races of Fusarium oxysporum f. sp. ciceris

2006 ◽  
Vol 84 (9) ◽  
pp. 1395-1404 ◽  
Author(s):  
Inmaculada Jorge ◽  
Juan A. Navas-Cortés ◽  
Rafael M. Jiménez-Díaz ◽  
Manuel Tena
Keyword(s):  
Cell Wall ◽  
Fusarium Oxysporum ◽  
Fusarium Wilt ◽  
Specific Activity ◽  
Gel Filtration ◽  
Pectate Lyase ◽  
Disease Development ◽  
In Planta ◽  
Cell Wall Degrading Enzymes ◽  
Degrading Enzymes

Production of cell wall degrading enzymes (CWDEs) polygalacturonase (PG), pectate lyase (PL), and xylanase was studied in chickpeas ( Cicer arietinum L. ‘P-2245’) inoculated with Fusarium oxysporum f. sp. ciceris (Padwick) Matuo & K. Sato races 0 (mildly virulent, causing a yellowing syndrome) and 5 (highly virulent, causing a wilting syndrome) by the water-culture method. These CWDEs were similarly produced in both syndromes. PG and PL were the only enzymes occurring in roots and stems and attained the highest specific activity, this being generally higher for race 5 than for race 0. Gel filtration chromatography revealed a similar complement of in planta expressed pectinase isoforms, dominated by an endo-PG and two endo-PLs, the endo-PLs being differentially expressed by the two races. CWDE activities in roots and stems were positively correlated with development of yellowing and wilting. Exceptions to this were PG in stems, which was negatively correlated with the development of yellowing, and PG in roots, which showed a negative trend with development of either syndrome. The levels of CWDEs that significantly correlated with disease development were adequately described by exponential functions of disease progress. Results have implications for the role played by CWDEs in the early and later stages of pathogenesis in chickpea fusarium wilt.

scholarly journals Race 3, a New and Highly Virulent Race of Fusarium oxysporum f. sp. niveum Causing Fusarium Wilt in Watermelon

Plant Disease ◽  
2010 ◽  
Vol 94 (1) ◽  
pp. 92-98 ◽  
Author(s):  
X. G. Zhou ◽  
K. L. Everts ◽  
B. D. Bruton
Keyword(s):  
Fusarium Oxysporum ◽  
Host Range ◽  
Fusarium Wilt ◽  
Citrullus Lanatus ◽  
Genetic Difference ◽  
Infested Soil ◽  
Reference Isolate ◽  
New Race ◽  
Race 2 ◽  
Highly Virulent

Three races (0, 1, and 2) of Fusarium oxysporum f. sp. niveum have been previously described in watermelon (Citrullus lanatus) based on their ability to cause disease on differential watermelon genotypes. Four isolates of F. oxysporum f. sp. niveum collected from wilted watermelon plants or infested soil in Maryland, along with reference isolates of races 0, 1, and 2, were compared for virulence, host range, and vegetative compatibility. Race identification was made on the watermelon differentials Sugar Baby, Charleston Gray, Dixielee, Calhoun Gray, and PI-296341-FR using a root-dip, tray-dip, or pipette inoculation method. All four Maryland isolates were highly virulent, causing 78 to 100% wilt on all differentials, one of which was PI-296341-FR, considered highly resistant to race 2. The isolates also produced significantly greater colonization in the lower stems of PI-296341-FR than a standard race 2 reference isolate. In field microplots, two of the isolates caused over 90% wilt on PI-296341-FR, whereas no disease was caused by a race 2 isolate. All four isolates were nonpathogenic on muskmelon, cucumber, pumpkin, and squash, confirming their host specific pathogenicity to watermelon. The Maryland isolates were vegetatively compatible to each other but not compatible with the race 2 isolates evaluated, indicating their genetic difference from race 2. This study proposes that the Maryland isolates belong to a new race, race 3, the most virulent race of F. oxysporum f. sp. niveum described to date.

scholarly journals First Report of Fusarium Wilt of Cilantro Caused by Fusarium oxysporum in California

Plant Disease ◽  
2005 ◽  
Vol 89 (10) ◽  
pp. 1130-1130 ◽  
Author(s):  
S. T. Koike ◽  
T. R. Gordon
Keyword(s):  
Fusarium Oxysporum ◽  
Fusarium Wilt ◽  
Vascular Tissue ◽  
Fusarium Species ◽  
Wilt Disease ◽  
Apium Graveolens ◽  
Peat Moss ◽  
State University ◽  
First Report ◽  
Vascular Discoloration

Cilantro, or coriander (Coriandrum sativum), is a leafy vegetable in the Apiaceae and is grown commercially in California primarily for use as a fresh herb. During 2002 and 2003 in coastal California (Santa Barbara County), commercial cilantro fields showed symptoms of a wilt disease. Affected plants grew poorly and were stunted. Lower foliage turned yellow with reddish tinges, and plants wilted during warmer times of the day. The main stem, crown, and taproot exhibited vascular discoloration that was reddish to light brown. As disease progressed, plants eventually died. For both years, the disease distribution was limited to isolated small patches (each patch measuring less than 1 m2 in area). A fungus was consistently isolated from symptomatic vascular tissue in crowns and taproots. On the basis of colony and conidial morphology, the isolates were identified as Fusarium oxysporum (2). No other fungi or bacteria were recovered from these plants. To test pathogenicity, suspensions containing 1 × 106 conidia/ml were prepared for five isolates. The roots of 30-day-old cilantro plants of four cultivars (30 plants each of Festival, Leisure, Santo, and LSO 14) were clipped and then soaked in the suspensions for 20 min. The roots of 30 plants of each cultivar were soaked in water as a control. Plants were repotted into new redwood bark + peat moss rooting medium and maintained in a greenhouse setting at 24 to 26°C. After 1 month, 95% or more of the inoculated plants showed yellowing and vascular discoloration symptoms similar to those seen in the field. F. oxysporum was reisolated from all inoculated plants. The four cilantro cultivars did not show differences in disease severity. Control plants showed no symptoms, and the fungus was not recovered from these plants. The experiment was repeated and the results were the same. Experiments also were conducted to determine if cilantro isolates could cause disease in celery (Apium graveolens var. dulce). Celery transplants and cilantro seedlings were prepared and inoculated as described above. However, after 2 months, celery plants did not show any disease symptoms, while the cilantro developed wilt symptoms and eventually died. A Fusarium wilt disease has been reported on coriander in Argentina and India where the pathogen was named F. oxysporum f. sp. coriandrii (1,3). To our knowledge, this is the first report of Fusarium wilt of cilantro in California. References: (1) M. Madia et al. Fitopatologia 34:155, 1999. (2) P. E. Nelson et al. Fusarium species: An Illustrated Manual for Identification. Pennsylvania State University Press, University Park, 1983. (3) U. S. Srivastava. Indian Phytopathol. 22:406, 1969.

scholarly journals Antagonisme Bacillus terhadap Infeksi Layu Fusarium pada Bibit Pisang Hasil Kultur Jaringan

2013 ◽  
Vol 15 (1) ◽  
pp. 21
Author(s):  
Hadiwiyono Hadiwiyono ◽  
Arief Widyantoro ◽  
Salim Widono
Keyword(s):  
Biological Control ◽  
Fusarium Oxysporum ◽  
Fusarium Wilt ◽  
In Planta ◽  
Physiological Character ◽  
Important Disease

<p>Fusarium wilt (Fusarium oxysporum f.sp. cubense) is an important disease in banana. Fusarium wilt was hard to control because the pathogen can survive in many kind of soils type although there is no host. Therefore, overcoming the disease is urgently needed such as biological control. The endophytic Bacillus of banana was begun to use as antagonist agent to the pathogen. This research aimed to study the mechanism of antagonism and physiological character of Bacillus. There were 27 Bacillus isolates examined in-vitro to test the production of IAA, HCN, chitinase, pectinase, and antagonism. The top ten isolates based on the test in vitro were used to test in planta. The research showed that Bacillus were able to produce IAA, HCN, chitinase, pectinase, and able to retard the growth of Foc colony. The application of isolat B25 on banana seedling could decrease the disease intensity but still unable to prevent the fusarium wilt infection.</p>

scholarly journals A DNA-Based Procedure for In Planta Detection of Fusarium oxysporum f. sp. phaseoli

2002 ◽  
Vol 92 (3) ◽  
pp. 237-244 ◽  
Author(s):  
Fernando M. Alves-Santos ◽  
Brisa Ramos ◽  
M. Asunción García-Sánchez ◽  
Arturo P. Eslava ◽  
José María Díaz-Mínguez
Keyword(s):  
Common Bean ◽  
Fusarium Oxysporum ◽  
Scar Marker ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
In Planta ◽  
Sequence Characterized Amplified Region ◽  
Polymerase Chain ◽  
Highly Virulent

We have characterized strains of Fusarium oxysporum from common bean fields in Spain that were nonpathogenic on common bean, as well as F. oxysporum strains (F. oxysporum f. sp. phaseoli) pathogenic to common bean by random amplified polymorphic DNA (RAPD) analysis. We identified a RAPD marker (RAPD 4.12) specific for the highly virulent pathogenic strains of the seven races of F. oxysporum f. sp. phaseoli. Sequence analysis of RAPD 4.12 allowed the design of oligonucleotides that amplify a 609-bp sequence characterized amplified region (SCAR) marker (SCAR-B310A280). Under controlled environmental and greenhouse conditions, detection of the pathogen by polymerase chain reaction was 100% successful in root samples of infected but still symptomless plants and in stem samples of plants with disease severity of ≥4 in the Centro Internacional de Agricultura Tropical (CIAT; Cali, Colombia) scale. The diagnostic procedure can be completed in 5 h and allows the detection of all known races of the pathogen in plant samples at early stages of the disease with no visible symptoms.

scholarly journals New Virulence Groups in Fusarium oxysporum f. sp. phaseoli: The Expression of the Gene Coding for the Transcription Factor ftf1 Correlates with Virulence

2011 ◽  
Vol 101 (4) ◽  
pp. 470-479 ◽  
Author(s):  
José J. de Vega-Bartol ◽  
Raúl Martín-Dominguez ◽  
Brisa Ramos ◽  
María-Asunción García-Sánchez ◽  
José María Díaz-Mínguez
Keyword(s):  
Transcription Factor ◽  
Common Bean ◽  
Fusarium Oxysporum ◽  
Quantitative Polymerase Chain Reaction ◽  
In Planta ◽  
Gene Encoding ◽  
Runner Bean ◽  
Bean Plants ◽  
Highly Virulent ◽  
Common Bean Plants

Fusarium oxysporum f. sp. phaseoli strains isolated from runner bean plants showing Fusarium wilt symptoms were characterized. The analysis of the genetic diversity of these strains and the comparison with strains formerly isolated from diseased common bean plants grown in the same region of Spain indicated a close genetic similarity among them. Pathogenicity assays carried out on runner bean plants showed virulence differences that allowed the classification of these strains into three groups: super virulent, highly virulent, and weakly virulent. However, all the analyzed strains behaved as highly virulent when inoculated on common bean plants, indicating that virulence is specific of the host–pathogen interaction. We also analyzed the number of copies and expression of the gene encoding the transcription factor ftf1, which has been shown to be specific of virulent F. oxysporum strains and highly up-regulated during plant infection. In planta real-time quantitative polymerase chain reaction expression analysis showed that expression of ftf1 was correlated with the degree of virulence. The comparative analysis of the polymorphic copies of ftf1 detected in the strains here characterized and those detected in the genome sequence of F. oxysporum f. sp. lycopersici strain 4287 indicates that some of the copies are likely nonfunctional.

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