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荧光定量PCR方法检测白酒发酵过程中Aspergillus tubingensis生物量 被引量:3

Detection and quantification the biomass of Aspergillus tubingensis in Chinese liquor fermentation process by real-time qPCR
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摘要 【目的】为了更好地分析霉菌在白酒发酵过程中的作用,需要快速准确地测定发酵过程中霉菌生物量的变化,本实验以白酒酿造中常用的塔宾曲霉(Aspergillus tubingensis)为例,建立一套快速准确定量塔宾曲霉生物量的方法。【方法】优化从酒醅中提取基因组的方法,设计和验证专一性引物,建立实时荧光定量PCR(Real-time quantitative PCR)方法,验证方法的有效性并应用于白酒发酵过程中塔宾曲霉生物量的检测。【结果】用原位机械破碎法提取酒醅中总基因组,其DNA的浓度能够达到1.060×105 ng/g酒醅;同时建立了一套快速准确测定固态基质中霉菌生物量的方法,并应用于白酒生产(制曲、堆积发酵和窖池发酵过程)中塔宾曲霉生物量的定量。【结论】实时荧光定量PCR方法能够快速准确地测定固态基质中霉菌的生物量,且检测限较低,对今后的相关研究具有借鉴意义。 [Objective] In order to better understand the role of filamentous fungi in Chinese liquor fermentation process, a rapid and accurate method to monitor the change of fungal biomass is necessary. This study established a real-time q PCR method to detect and quantify Aspergillus tubingensis, which is widely used in Chinese liquor fermentation. [Methods] The methods of extracting genome from fermented grains have been optimized, the specific primers for A. tubingensis has also been designed and validated. [Results] The total DNA extracting from fermented grains can reach 1.060×105 ng/g by in situ mechanical crushing method. The applicability of real-time q PCR method to detect A. tubingensis in solid-state matrix has been validated in liquor fermentation process. [Conclusion] Real-time q PCR method can rapidly and accurately detect the fungal biomass in solid-state matrix, which provides a powerful tool for related researches.
出处 《微生物学通报》 CAS CSCD 北大核心 2014年第12期2547-2554,共8页 Microbiology China
基金 国家863计划项目(No.2012AA021301,2013AA102108) 国家自然科学基金项目(No.31000806,31371822,31271921) 2011协同创新计划
关键词 荧光定量PCR 霉菌 固态发酵 塔宾曲霉 生物量 Real-time qPCR,Filamentous fungi,SSF,Aspergillus tubingensis,Biomass
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  • 1李海梅,马莺.曲霉菌酶学性质的研究[J].中国酿造,2004,23(7):6-9. 被引量:5
  • 2高修功,章克昌.纤维素酶固态发酵过程中菌体生长量的测定[J].工业微生物,1994,24(3):26-30. 被引量:15
  • 3Haruta S, Ueno S, Egawa I, et al. Succession of bacterial and fungal communities during a traditional pot fermentation of rice vinegar assessed by PCR-mediated denaturing gradient gel electrophoresis. International Journal of Food Microbiology, 2006, 109 : 79-87.
  • 4Aquilanti L, Santarelli S, Silvestri G, et al. The microbial ecology of a typical Italian salami during its natural fermentation. International Journal of Food Microbiology, 2007, 120 (1-2): 136-145.
  • 5Ercolini D, Hill PJ, Dodd CER. Bacterial community structure and location in Stilton cheese. Applied and Environmental Microbiology, 2003, 69 (6): 3540-3548.
  • 6Nanda K, Taniguchi M, Ujike S, et al. Characterization of acetic acid bacteria in traditional acetic acid fermentation of rice vinegar (komesu) and unpolished rice vinegar (kurosu) produced in Japan. Applied and Environmental Microbiology, 2001, 17: 986-990.
  • 7Di Maro E, Ercolini D, Coppola S. Yeast dynamics during spontaneous wine fermentation of the Catalanesca grape. International Journal of Food Microbiology, 2007, 117 (2) : 201-210.
  • 8Prieto C, Jara C, Mas A, et al. Application of molecular methods for analysing the distribution and diversity of acetic acid bacteria in Chilean vineyards. International Journal of Food Microbiology, 2007, 115 ( 3 ): 348-355.
  • 9de Lipthay JR, Enzinger C, Johnsen K, et al. Impact of DNA extraction method on bacterial community composition measured by denaturing gradient gel electrophoresis. Soil Biology and Biochemistry, 2004, 36 (10) : 1607-1614.
  • 10Demeke T, Adams RP. The effects of plant polysaccharides and buffer additives on PCR. BioTechniques, 1992, 12: 332-334.

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