Quantitative Detection of Bacillus subtilis HMB19198 on Tomato Leaf by Real-time PCR

doi:10.16409/j.cnki.2095-039x.2022.02.010

Abstract

Abstract: The biocontrol agent Bacillus subtilis HMB19198 is proved to be effective in control of tomato gray mold in the field.To evaluate the colonization ability of strain HMB19198 on tomato leaf surface，a specific gene of strain HMB19198 was obtained by BLAST of all genes in HMB19198, followed by designing a pair of the strain-specific primers and DNA probe. Real-time PCR (qPCR) experiments showed that the primers and probe were efficient in specific detection of strain HMB19198 in laboratory and the detection sensitivity was 10²copies/μL when using HMB19198 DNA spiked with tomato leaf DNA. Combing the qPCR technique and the colony-counting method (CCM), the colonization dynamics of strain HMB19198 on tomato leaves were monitored. Results showed that 0 day after inoculation (DAI), the population of strain HMB19198 on the leaf was detected as 1.0×10⁷ copies/g leaf by qPCR and 8.9×10⁷ cfu/g leaf by CCM. The population of strain HMB19198 on the leaf was gradually decreased at 4, 6 and 8 DAI, and the population at 8 DAI was detected as 1.0×10⁷ copies/g leaf by qPCR and 1.2×10⁷ cfu/g by CCM. The control efficacy was above 80% againsr tomato gray mold after application of strain HMB19198 for 2 days, while the efficacy decreased to 37.9% after 8 days.

Key words: Bacillus subtilis, real-time PCR, tomato gray mold, colonization

CLC Number:

S476

LIU Xiaomeng, SU Zhenhe, XUAN Lifeng, WANG Peipei, DONG Lihong, GUO Qinggang, MA Ping. Quantitative Detection of Bacillus subtilis HMB19198 on Tomato Leaf by Real-time PCR[J]. Chinese Journal of Biological Control, 2022, 38(2): 487-494.

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