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| Establishment of a Rapid qPCR Detection Method for Burkholderia stagnalis |
| DONG Jin-Bao1, DENG Ying-Long1, YANG Bi-Chen1, ZHANG Ying-Long1, HUANG Hong-Ping1, ZHU You-Yong1, HE Xia-Hong1,2,*, GUO Li-Wei1,* |
1 State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming 650201, China;
2 Provincial Key Laboratory for Conservation and Utilization of In-forest Resource, Southwest Forestry University, Kunming 650224, China |
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Abstract A strain of Burkholderia stagnalis B11 was previously isolated by our research group, which exhibits an effective biocontrol effect against root rot disease of Panax notoginseng. To establish a rapid detection method based on qPCR for quantifying the colonization capacity of strain B11 in soil, a comparative genomic analysis was conducted. A specific gene, gene 1214 (681 bp), located within the MIBiG cluster BGC0001120 and annotated as homolog of burkholderic acid synthesis genes, was selected as the target. Specific primers targeting strain B11 were designed using NCBI Primer-BLAST. qPCR results showed that the designed primers exhibited good specificity. The established standard curve equation was y=-3.3689x+11.343 (R2=0.9991), with an amplification efficiency of 98.08% and a sensitivity of 0.02 pg/µL. This method was used to detect the colonization of B. stagnalis B11 in continuous cropping soil of P. notoginseng. The result showed that the abundance of B11 in the soil decreased with the increase of the bacterial suspension dilution ratio. A significant decrease occurred at a 10-fold dilution, and no significant difference compared with the control (CK) was observed at a 100-fold dilution. The colonization dynamics exhibited an initial increase followed by a subsequent decrease, with stable colonization maintained in the soil from 1 to 5 d after irrigation. This study established a qPCR method that enabled rapid quantitative detection of B. stagnalis B11, which provides a reliable technical means for evaluating the colonization capacity of this strain in the soil environment and facilitating the development of an efficient detection system.
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Received: 09 July 2025
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Corresponding Authors:
*hexiahong@hotmail.com; g87l12w12@163.com
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