Establishment of Real-time Fluorescent Quantitative PCR for Trichoderma brevicompactum and Its Dynamic Change in Pumpkin Rhizosphere

doi:10.16409/j.cnki.2095-039x.2018.04.012

Abstract

Abstract: A real-time fluorescent quantitative PCR detection assay of Trichoderma brevicompactum 31636 was developed based on a specific primer pair TB51F/51R which were designed based on the ITS gene. The pot experiment was conducted to determine the control effect of T. brevicompactum 31636 on pumpkin Phytophthora blight, and the dynamic changes of population of T. brevicompactum 31636 and Phytophthora capsici Leonian LJ12010805 in pumpkin rhizosphere soil were monitored by RT-PCR. The result showed that the assay has a good liner relationship with a high sensitivity of 0.16 pg/μL DNA, which is 100 times more than that of normal PCR. Inoculation with 10 mL T. brevicompactum 31636 bacterial suspension at sowing stage had a best control effect on pumpkin Phytophthora blight, with the efficacies of 73.11% and 46.82% on the 15th and 30th day, respectively. RT-PCR results showed that dynamic change of T. brevicompactum 31636 in matrix after planting was appeared three distinct phases that were decrease, increase and decrease. The amount of bacteria was the largest on 15th day, with the copy number of 3.44×10⁶ copys/μL. On the 3rd after planting, the amount of P. capsici LJ12010805 reached the maximum, then decreased gradually and reached the lowest on 15th day, with the copy number of 2.42×10⁶ copys/μL and 5.76×10²copys/μL, respectively. However, P. capsici LJ12010805 of the control showed an increasing trend, with the highest copy number of 8.42×10⁷copys/μL on the 30th day.

Key words: Trichoderma brevicompactum, pumpkin Phytophthora blight, real-time fluorescent quantitative PCR, dynamic change, control effect

CLC Number:

S476

XIE Xuewen, CHENG Yingchao, WANG Kun, ZHANG Hongjie, YANG Jie, SHI Yanxia, CHAI Ali, LI Baoju. Establishment of Real-time Fluorescent Quantitative PCR for Trichoderma brevicompactum and Its Dynamic Change in Pumpkin Rhizosphere[J]. journal1, 2018, 34(4): 574-581.

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