scholarly journals Pre-treatment and Suitable Reagent Enabled a Reliable and Consistent for Molecular Detection of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc4)

2019 ◽  
Vol 26 (4) ◽  
pp. 196
Author(s):  
Listya Utami Karmawan ◽  
Fenny Martha Dwivany ◽  
Rizkita Rachmi Esyanti ◽  
I Nyoman Pugeg Aryantha

Fusarium wilt which is caused by the soil-borne fungus, Fusarium oxysporum f.sp. cubense (Foc), is one of important diseases in banana plant. Foc tropical race 4 (Foc4) is the most pathogenic race which could infect various banana cultivars including Cavendish cultivar which was previously considered as resistant cultivar. Molecular detection of Foc using PCR analysis is indispensable to determine the race of Foc. We demonstrate a faster DNA isolation procedure described in previous method by substituting sodium acetate precipitation with ammonium acetate precipitation without affecting the result. Based on our experience, some fungal genomes were troublesome to be amplified. We suggested pre-treatment step prior to amplification procedure by incubating fungal DNA in 65°C for 10 minutes for any samples of fungal genome, including stubborn samples, before mixing into PCR mix reagent. PCR reagents should be tested for stubborn samples since some of the reagents were unable to amplify the desired DNA fragment. Pre-treatment and the choice of robust PCR reagent should be taken into consideration for a reliable and consistent Foc4 molecular detection result.

PLoS ONE ◽  
2019 ◽  
Vol 14 (9) ◽  
pp. e0222727 ◽  
Author(s):  
Freddy Magdama ◽  
Lorena Monserrate-Maggi ◽  
Lizette Serrano ◽  
Daynet Sosa ◽  
David M. Geiser ◽  
...  

1995 ◽  
Vol 46 (1) ◽  
pp. 167 ◽  
Author(s):  
KG Pegg ◽  
RG Shivas ◽  
NY Moore ◽  
S Bentley

A unique population of Fusarium oxysporum f. sp. cubense affecting Cavendish cv. Williams banana plants was characterized using vegetative compatibility, volatile production, RAPD-PCR analysis, pectic enzyme production and pathogenicity. The isolates were more like race 1 isolates than race 4 isolates, although they were capable of attacking Cavendish clones. The Carnarvon isolates did not belong to any of the vegetative compatibility groups (VCGs) known to occur in Australia or overseas; they belonged in the 'inodoraturn' volatile group; they had 29% genetic similarity to race 4 isolates and 76% similarity to race 1 isolates based on RAPD-PCR banding patterns; they belonged in the same pectic zymogram group as race 1 isolates and were virulent on 3-month-old Cavendish cv. Williams, Gros Michel and Pisang Gajih Merah plants in glasshouse tests.


Biomolecules ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1353
Author(s):  
Yanqiu He ◽  
Xiaofan Zhou ◽  
Jieling Li ◽  
Huaping Li ◽  
Yunfeng Li ◽  
...  

Banana Fusarium wilt, caused by the fungus pathogen Fusarium oxysporum f. sp. cubense (Foc), is a devastating disease that causes tremendous reductions in banana yield worldwide. Secreted proteins can act as pathogenicity factors and play important roles in the Foc–banana interactions. In this study, a shotgun-based proteomic approach was employed to characterize and compare the secretomes of Foc1 and Foc4 upon banana extract treatment, which detected 1183 Foc1 and 2450 Foc4 proteins. Comprehensive in silico analyses further identified 447 Foc1 and 433 Foc4 proteins in the classical and non-classical secretion pathways, while the remaining proteins might be secreted through currently unknown mechanisms. Further analyses showed that the secretomes of Foc1 and Foc4 are similar in their overall functional characteristics and share largely conserved repertoires of CAZymes and effectors. However, we also identified a number of potentially important pathogenicity factors that are differentially present in Foc1 and Foc4, which may contribute to their different pathogenicity against banana hosts. Furthermore, our quantitative PCR analysis revealed that genes encoding secreted pathogenicity factors differ significantly between Foc1 and Foc4 in their expression regulation in response to banana extract treatment. To our knowledge, this is the first experimental secretome analysis that focused on the pathogenicity mechanism in different Foc races. The results of this study provide useful resources for further exploration of the complicated pathogenicity mechanisms in Foc.


Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 670
Author(s):  
Xiaoxia Zhang ◽  
Huoqing Huang ◽  
Bangting Wu ◽  
Jianghui Xie ◽  
Altus Viljoen ◽  
...  

Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) causes Fusarium wilt of banana, the most devastating disease on a banana plant. The genome of Foc TR4 encodes many candidate effector proteins. However, little is known about the functions of these effector proteins on their contributions to disease development and Foc TR4 virulence. Here, we discovered a secreted metalloprotease, FocM35_1, which is an essential virulence effector of Foc TR4. FocM35_1 was highly upregulated during the early stages of Foc TR4 infection progress in bananas. The FocM35_1 knockout mutant compromised the virulence of Foc TR4. FocM35_1 could interact with the banana chitinase MaChiA, and it decreased banana chitinase activity. FocM35_1 induced cell death in Nicotiana benthamiana while suppressing the INF1-induced hypersensitive response (HR), and its predicted enzymatic site was required for lesion formation and the suppression to INF1-induced HR on N. benthamiana leaves. Importantly, treatment of banana leaves with recombinant FocM35_1 accelerates Foc TR4 infection. Collectively, our study provides evidence that metalloprotease effector FocM35 seems to contribute to pathogen virulence by inhibiting the host immunity.


2013 ◽  
Vol 38 (2) ◽  
pp. 173-176 ◽  
Author(s):  
Yong-hong HUANG ◽  
Shun LÜ ◽  
Chun-yu LI ◽  
Yue-rong WEI ◽  
Gan-jun YI

Plant Disease ◽  
2018 ◽  
Vol 102 (2) ◽  
pp. 448-448 ◽  
Author(s):  
T. N. Hung ◽  
N. Q. Hung ◽  
D. Mostert ◽  
A. Viljoen ◽  
C. P Chao ◽  
...  

2018 ◽  
Vol 48 (6) ◽  
Author(s):  
Marcelo Marques da Silveira ◽  
Stéfhano Luis Cândido ◽  
Karin Rinaldi dos Santos ◽  
Maerle Oliveira Maia ◽  
Roberto Lopes de Souza ◽  
...  

ABSTRACT: Sepsis is characterized by the presence of organ dysfunction secondary to the dysregulated systemic inflammatory response associated with an infection, and has high mortality rates. Traditional diagnostic techniques based on non-microbiological isolation are time-consuming and may delay treatment. Thus, this study aimed to compare bacterial and fungal broad-range polymerase chain reaction (PCR) and blood culture for diagnosis of sepsis in dogs. Blood samples from 88 dogs with suspected sepsis were analyzed by blood culture, and PCR to detect bacterial and fungal DNA. On blood culture, 20 (22.7%) samples tested positive for bacterial isolates; however, none tested positive for fungi. Through PCR analysis, bacterial DNA was detected in 46 (52.3%) animals, whereas fungal DNA was present in one (1.1%) sample. Our results showed that PCR-based testing has important diagnostic value for canine blood infections because it has a shorter turnaround time and higher sensitivity than traditional blood culture.


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