Rapid Detection and Quantification of Viable Cells of Pectobacterium brasiliense Using Propidium Monoazide Combined with Real-Time PCR
文献类型： 外文期刊
作者： Li, Junhui 1 ; Chen, Ruxing 1 ; Yang, Ruwei 2 ; Wei, Xinchen 1 ; Xie, Hua 3 ; Shi, Yanxia 1 ; Xie, Xuewen 1 ; Chai, Ali 1 ; Fan, Tengfei 1 ; Li, Baoju 1 ; Li, Lei 1 ;
作者机构： 1.Chinese Acad Agr Sci, Inst Vegetables & Flowers, State Key Lab Vegetable Biobreeding, Beijing 100081, Peoples R China
2.Xinjiang Acad Agr Sci, Comprehens Expt Farm, Urumqi 830091, Peoples R China
3.Beijing Acad Agr & Forestry Sci, Beijing Agrobiotechnol Res Ctr, Beijing 100097, Peoples R China
关键词： Pectobacterium brasiliense; bacterial soft rot; PMA-qPCR; detection; viable cell
期刊名称：MICROORGANISMS （ 影响因子：4.5； 五年影响因子：4.8 ）
ISSN：
年卷期： 2023 年 11 卷 11 期
页码：
收录情况： SCI
摘要：
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; R-2 = 0.99). The amplification efficiency reached 97%, and the lowest detection limit of viable cells was approximately 2 x 10(2) CFU
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