Development and Application of Quantitative PCR for Detection of Fusarium

doi:10.7668/hbnxb.20190656

Abstract

Abstract: To establish a method for rapid detection of Fusarium. Real-time PCR assay for quantitative detection of Fusarium in cucumber was developed and a pair of specific primers were designed based on the genome sequence of cucumber Fusarium and a target fragment of 187 bp was specifically amplified from the genomic DNA of Fusarium. The detection sensitivity of RT-PCR established in this study was 10⁴ times higher than that of conventional PCR. RT-PCR method was used to investigate the variation of crop residues DNA and residues DNA of soil in different humidity. The results showed that the initial copy numbers of DNA of crop residues and crop residues of soil was 6.90×10¹¹,1.06×10¹² copies/g,respectively. After treatment at temperature 27 ℃ and 80% humidity for 30 days, the DNA content of the crop residues decreased to 5.55×10⁸ and 0 copies/g,respectively. The content was 27 ℃ and 20% humidity was 8.04×10⁹ and 1.30×10⁹ copies/g,respectively. Therefore, these studies showed that RT-PCR assay is a highly rapid and reliable method to quantify Fusarium in cucumber crop residues. Application of the assay may potentially improve pathogen identification and disease management.

Key words: Fusarium, Real-time PCR, Humidity, Crop residues

CLC Number:

S432.4

CHEN Lida, YUAN Junhai, LI Lei, SHI Yanxia, CHAI Ali, XIE Xuewen, LI Baoju. Development and Application of Quantitative PCR for Detection of Fusarium[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(S1): 296-301. doi: 10.7668/hbnxb.20190656.

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