JIA-2018-09

2130 WANG Ying et al. Journal of Integrative Agriculture 2018, 17(9): 2126–2136 and DQ104727.1). Based on this result, Hmp-1 was classi ed as a member of P . membranifaciens . 3.4. Population dynamics of FM-S-115 and Hmp-1 during blackberry juice fermentation To analyze the growth dynamics and competitive position of FM-S-115 and Hmp-1, their population densities were investigated when they were inoculated solely or jointly into blackberry juice (Fig. 4). The population density of FM-S- 115 increased about 100 times within 6 d after inoculation, and then decreased. The population density of Hmp-1 always decreased after alone inoculation, and then the living cells were not detected at 8–14 d. Compared to the growth dynamics of single inoculation, the growth dynamics of FM-S-115 was unchanged at 1–6 d after inoculation in the joint inoculation blackberry juice, and then the cells density was lower than that of single inoculation blackberry juice. Moreover, we also found that the cells density of Hmp-1 decreased at 1–6 d after inoculation, and then sharply increased in joint inoculation blackberry juice, which was a Pichia membranifaciens (JX188208) Pichia membranifaciens (DQ198957) Pichia membranifaciens (EF564396) Hmp-1 (KJ914549) (this study) Pichia membranifaciens (DQ104727) Candida ethanolica (FM180545) Candida ethanolica (HG425336) Pichia manshurica (JF912077) Pichia manshurica (FM180542) Pichia manshurica (FR870032) Pichia manshurica (JF912079) Pichia kudriavzevii genes (AB617989) Schizosaccharomyces pombe (HE964968) Zygosaccharomyces mrakii (U72159) Zygosaccharomyces rouxii (AB363045) Saccharomyces cerevisiae (AB733454) Saccharomyces cerevisiae FM-S-115 (KM408172) Kluyveromyces lactis (HE660074) 100 99 90 97 95 93 74 69 65 0.1 Fig. 3 The phylogenetic tree of Pichia membranifaciens Hmp-1 was constructed using neighbor-joining method based on the partial nucleotide sequence of 26S rDNA D1/D2 domain from yeasts related to Pichia species. Numbers at the branches denote the bootstrap percentages for 1000 replicates. KJ914549 in parentheses indicates the accession numbers deposited in the GenBank database in this study, and the accession numbers for reference sequences are shown in parenthesis. The sequence of 26S rDNA D1/D2 domain from Kluyveromyces lactis (HE660074.1) was used as an outgroup. The scale indicates the evolutionary distance of the nucleotide substitutions per site. 0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 0 3 6 9 12 15 Log 10 CFU mL –1 Fermentation time (d) Fig. 4 Population dynamics of Pichia membranifaciens Hmp-1 and Saccharomyces cerevisiae FM-S-115 strain in blackberry juice during fermentation. Blackberry juice was fermented at 28°C for 14 d. , population dynamics of Hmp-1 in the blackberry juice inoculated solely; ■, population dynamics of Hmp-1 in the blackberry juice jointly inoculated with S. cerevisiae FM-S-115; , population dynamics of S. cerevisiae FM-S-115 in the blackberry juice jointly inoculated with Hmp-1; , population dynamics of S. cerevisiae FM-S-115 in the blackberry juice inoculated solely. Vertical bars represent the standard errors of the means.