TH5778Y - A primer set for identifying the race of the fungus Fusarium oxysporum f. sp. lycopersici, the causative agent of tomato yellow wilt disease. - Google Patents

Abstract

The DC60 primer set was used to detect the race of the fungus Fusarium oxysporum f. sp. lycopersici, the causative agent of tomato wilt disease. The set consisted of four primer pairs: uni (forward) + uni (reverse), tom3 (forward) + tom3 (reverse), B20 (forward) + B20 (reverse), and B35 (forward) + B35 (reverse) in a polymerase chain reaction (PCR) with the following steps: Step 1: 95°C for 7 minutes, 1 cycle; Step 2: 94°C for 1 minute; 55°C for 1.30 minutes; 72°C for 2 minutes, 30 cycles; Step 3: 72°C for 5 minutes, 1 cycle. The PCR products were then size-separated using 2% agarose gel electrophoresis. DNA fragments of Fol race 1, 2, and 3 were obtained as follows: For Fol race 1, the DNA fragment was obtained from the 670-base pair uni primer, the 458-base pair tom3 primer, the 283-base pair B20 primer, and the 247-base pair B35 primer. For Fol race 2, the DNA fragment was obtained from the 670-base pair uni primer and the 247-base pair B35 primer. For Fol race 3, the DNA fragment was obtained from the 670-base pair uni primer and the 458-base pair tom3 primer. This primer set was used to identify the race of the fungus Fusarium oxysporum f. sp. lycopersici, the causative agent of tomato wilt disease. This study used a primer set consisting of four pairs: uni (forward) + uni (reverse), tom3 (forward) + tom3 (reverse), B20 (forward) + B20 (reverse), and B35 (forward) - B35 (reverse) in an in vitro polymerase chain reaction (PCR) to amplify DNA fragments. The program was set up as follows: Step 1: 95°C for 7 minutes, 1 cycle; Step 2: 94°C for 1 minute; 55°C for 1.30 minutes; 72°C for 2 minutes, 30 cycles; Step 3: 72°C for 5 minutes, 1 cycle. The PCR product was then size-separated using 2% agarose gel electrophoresis. The DNA fragments of Fol race 1, 2, and 3 were obtained as follows: For Fol race 1, the DNA fragment was obtained from the 670-base pair uni primer, the 458-base pair tom3 primer, the 283-base pair B20 primer, and the 247-base pair B35 primer. For Fol race 2, the DNA fragment was obtained from the 670-base pair uni primer and the 247-base pair B35 primer. For Fol race 3, the DNA fragment was obtained from the 670-base pair uni primer and the 458-base pair tom3 primer.

Claims (4)

1. The primer set for detecting the race of the fungus Fusarium oxysporum f. sp. lycopersici, the causative agent of tomato wilt disease, consists of 4 pairs of primers with the following base sequences: -Primer pair 1: uni (forward) 5'- ATCATCTTGTGCCAACTTCAG-3' uni (reverse) 5'- GTTTGTGATCTTTGAGTTGCCA -3' -Primer pair 2: tom3 (forward) 5'- TTTCCGAGCAGTTCAACAGTCTTTC-3' tom3 (reverse) 5'- AGTTCTTGGCGCTTTTTCGGCAGTA-3' -Primer pair 3: B20 (forward) 5'- AATTCACTGCCGACGCCAG-3' B20 (reverse) 5'- TAACATATTTCGCCCTCATC-3' -Primer pair 4: B35 (forward) 5'- TAACTAACAAACAGTGAGAATG-3' B35 (reverse) 5'- AATTCACTACGCCCAGTAAAAG-3' 2. The primer set used to detect the race of the fungus Fusarium oxysporum f. sp. lycopersici, the causative agent of tomato wilt disease according to claim 1, specifically for Folrace 1, consists of a 670-base pair DNA fragment from the uni primer, a 458-base pair DNA fragment from the tom3 primer, a 283-base pair DNA fragment from the B20 primer, and a 247-base pair DNA fragment from the B35 primer. 3. The primer set used to detect the race of the fungus Fusarium oxysporum f. sp. lycopersici, the causative agent of tomato wilt disease according to claim 1, which is used to detect Folrace 2, will contain a DNA fragment from the uni primer of 670 base pairs and a DNA fragment from the B35 primer of 247 base pairs. 4. The primer set for detecting rase of the fungus Fusarium oxysporum f. sp. lycopersici, the causative agent of tomato wilt disease according to claim 1, which is used to detect the fungus Folrace 3, will contain a DNA fragment from the uni primer of 670 base pairs and a DNA fragment from the tom3 primer of 458 base pairs.