秸秆发酵乳酸菌复合系SFC-2的构建及其组成多样性研究

为了获得促进作物干秸秆乳酸发酵的微生物,以玉米秸秆和水稻秸秆的自然发酵物为菌种来源,用MRS蔗糖培养基,通过连续定向继代培养,筛选出pH下降迅速、乳酸含量高、组成稳定的乳酸菌复合系SFC-2.DGGE分析表明,SFC-2经过连续继代培养,从第25代开始其微生物组成基本稳定.SFC-2 培养12 h后pH下降至3.8,乳酸含量达10.64 mg/mL,其中64%为L-(+) 乳酸.通过平板分离获得4株细菌,全部为Lactobacillus,其近缘种分别为L. fermentum、L. plantarum、L. paracasei和L. paracasei sub sp.;通过16S rDNA克隆文库分析获得7个克隆,其近缘种主要为L. fermentum、L. plantarum及L.paracasei;在16S rDNA的克隆文库中,76.3%为L. fermentum的近缘种,20.3%为L. plantarum的近缘种,3.4%为L. paracasei的近缘种.

Construction and Composition Diversity of a Lactic Acid Bacterial Community SFC-2

To obtain the community that could accelerate the fermentation of the air-dried crop straws, a lactic acid bacteria community SFC-2 was constructed from the natural fermented products of the corn straw and rice straw by continuous restricted subcultivation in the MRS-S broth. The SFC-2 could lower the pH of the broth quickly, and produce high amount of lactic acid. The microbial composition of the SFC-2 became stable from the 25^th generation according to the results of denaturing gradient gel electrophoresis (DGGE). The pH value of the SFC-2 dropped to 3.8 and the amount of lactic acid reached 10.64 mg·mL^-1 after 12 h cultivation in the broth, which contained 64% L-(+) lactic acid. Using morphological methods, four strains were isolated from the SFC-2 and they all belonged to Lactobacillus. The closest species were L. fermentum, L. plantarum, L. paracasei and L. paracasei sub sp., respectively. At the same time, the composition of the SFC-2 was also analyzed by constructing 16S rDNA clone library. The results of 16S rDNA sequencing showed that the closest species of seven clones were mainly L. fermentum, L. plantarum and L. paracasei. In clone libraries, 76.3% belonged to L. fermentum, 20.3% belonged to L. plantarum and 3.4% belonged to L. paracasei.