Simple and sensitive BIO-PCR detection of potato blackleg pathogens from stem, tuber, and soil samples

Aono, Yoshiyuki; Nakayama, T.; Ozawa, Takahiro; Ushio, Yutaka; Yasuoka, Shinji; Fujimoto, Taketo; Ohki, Takehiro; Oka, Norikuni; Maoka, Tetsuo

doi:10.1007/s10327-021-00997-9

Cited by 4 publications

(1 citation statement)

References 21 publications

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“…However, the sensitivity of this method exhibits significant inaccuracies, as microorganisms in diseased seeds or plant tissues compete with or inhibit the growth of the target pathogen. Although a few reverse-transcription polymerase chain reaction (RT-PCR) studies have investigated Pectobacterium species [21,22], Pbr has not been studied.…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection and Quantification of Viable Cells of Pectobacterium brasiliense Using Propidium Monoazide Combined with Real-Time PCR

Li,

Chen,

Yang

et al. 2023

Microorganisms

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Pectobacterium brasiliense (Pbr) has caused significant economic losses in major vegetable production areas in Northern China by causing bacterial soft rot in cash crops such as potatoes and cucumbers. This study aimed to establish a PMA-qPCR detection method for Pbr by screening specific and sensitive primers based on the glu gene and the conserved region of the 23S rRNA gene. Based on the optimized PMA pretreatment conditions, a standard curve was designed and constructed for PMA-qPCR detection (y = −3.391x + 36.28; R2 = 0.99). The amplification efficiency reached 97%, and the lowest detection limit of viable cells was approximately 2 × 102 CFU·mL−1. The feasibility of the PMA-qPCR method was confirmed through a manually simulated viable/dead cell assay under various concentrations. The analysis of potato tubers and cucumber seeds revealed that nine naturally collected seed samples contained a range from 102 to 104 CFU·g−1 viable Pbr bacteria. Furthermore, the system effectively identified changes in the number of pathogenic bacteria in cucumber and potato leaves affected by soft rot throughout the disease period. Overall, the detection and prevention of bacterial soft rot caused by Pbr is crucial.

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Section: Introductionmentioning

confidence: 99%