商业酵母在葡萄酒工业化生产中的定殖情况分析

doi:10.3864/j.issn.0578-1752.2021.09.016

Abstract

Abstract:

【Objective】The purpose of this study was explore the implantation and persistence of commercial active dry yeast (ADY) during industrial wine fermentations, and their competitive relationship between Chinese indigenous Saccharomyces cerevisiae during fermentation process, so as to provide the theoretical basis for the breeding of indigenous S. cerevisiae strains and provide the reference for the use of ADY in wine production. 【Method】Industrial wine fermentations were carried out at wineries in Eastern Foot of Helan Mountain in Ningxia. Four vats of Cabernet Sauvignon gape must were inoculated with BDX, XR, FR and FX10, respectively. Samples were collected and analyzed at 1 d, 3 d and 5 d after the inoculation. Interdelta and SSR analysis were used to investigate the genotypes of different S. cerevisiae strains. Therefore, the number and proportion of S. cerevisiae strains in different fermentation stages were analyzed, and the colonization ability of commercial ADY was tracked. Genetic diversity parameters were calculated by PopGen32 software. The genetic correlation between commercial yeast and Ningxia indigenous yeast was revealed by NTsys2.10e software. 【Result】Interdelta fingerprint showed 6 kinds of fingerprints, namely 6 genotypes. And XR and FR showed more than one genotype; BDX and FX10 showed one genotype, respectively. SSR analysis showed that there was one genotype in each ADY for the 9 locus. 225 S. cerevisiae isolates were isolated from the 4 inoculated fermentations. Interdelta fingerprint showed 42 genotypes, of which 36 genotypes were indigenous strains. The degree of variability (16%, 42/225) was intermediate. SSR analysis showed 20 genotypes, of which 16 genotypes were indigenous strains. The analyzed 9 microsatellite prime pairs generated a total of 75 polymorphic bands, 8.3333 alleles for per locus. The heterozygosity observed was 0.2000-0.5000. The polymorphism information contents (PIC) of all strains at 9 loci were 0.6339-0.8620, suggesting that the 9 SSR loci were hypervariable. The indigenous genotypes were the most abundant in the fermentation inoculated with BDX (11 Interdelta types and 8 SSR types), followed by FR (11 Interdelta types and 6 SSR types). ADY did not dominate all three stages. Moreover, the genotypes of the dominant strains were also different for different stages in the same fermentation. Interdelta and SSR analysis showed FR was not dominant in the corresponding fermentation. Although BDX existed in the whole fermentation process, it was only dominant at d 3 after the inoculation. In the fermentation inoculated with XR and FX10, Interdelta analysis showed that they were not the dominant strains, while SSR analysis showed that they were the dominant strains in the corresponding fermentations, respectively. Indigenous strains of genotype β (SSR genotype BDX-7), genotype γ (SSR genotype BDX-6), genotype A (SSR genotype XR), genotype a (SSR genotype FX10), genotype b (SSR genotype FX10), genotype bb (SSR genotype FR-2) and genotype ee (SSR genotype FR-4) showed strong competitiveness in the corresponding fermentations. Cluster analysis showed that the genetic diversity among the S. cerevisiae strains isolated from the same fermentation was large. 【Conclusion】The genotypes of indigenous S. cerevisiae strains in the industrial wine fermentations were rich. The inoculated fermentations were completed by both indigenous strains and commercial ADY, and they competed with each other in the same fermentations and showed dynamic succession of different strains.

Key words: ADY, inoculated fermentation, strain typing, wine, SSR

SUN Yue,YANG HuiMin,HE RongRong,ZHANG JunXiang. Implantation and Persistence of Inoculated Active Dry Yeast in Industrial Wine Fermentations[J].Scientia Agricultura Sinica, 2021, 54(9): 2006-2016.

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References 32

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位点 Locus	染色体 Chromosome	重复序列 Repetitive sequences	引物序列 Primer sequence (5′-3′)	参考文献 Reference
SCAAT1	XIII	TTA	F: AAAGCGTAAGCAATGGTGTAGATACTT R: CAAGCCTCTTCAAGCATGACCTTT	[15-18]
YPL009C	XVI	CTT	F: AACCCATTGACCTCGTTACTATCGT R: TTCGATGGCTCTGATAACTCCATTC	[15,17]
C4	XV	TAA	F: AGGAGAAAAATGCTGTTTATTCTGACC R: TTTTCCTCCGGGACGTGAAATA	[15,17-18]
C5	Ⅵ	GT	F: TGACACAATAGCAATGGCCTTCA R: GCAAGCGACTAGAACAACAATCACA	[15,17-18]
C8	VII	TAA	F: CAGGTCGTTCTAACGTTGGTAAAATG R: GCTGTTGCTGTTGGTAGCATTACTGT	[15,18]
C11	X	GT	F: TGCGCAGCTTAGTATGACCA R: GATGGGCTTTCACTCCACTT	[16-18]
C12	XII	CAA	F: GAGGAGCTTACTTAAGAGCATGCGTTC R: GTGTCTTAAACCTATATTCGGATTGTGCCTGCT	[15]
SCAAT3	IV	AAT	F: TGGGAGGAGGGAAATGGACAG R: TTCAGTTACCCGCACAATCTA	[15,18]
SCYOR267C	XV	TGT	F: TACTAACGTCAACACTGCTGCCAA R: GGATCTACTTGCAGTATACGGG	[15,17]

ADY	SCAAT1	YPL009C	C4	C5	C8	C11	C12	SCAAT3	SCYOR267C
BDX	208	278	245	159	133	282	120	249	286
BDX	208	305	257	169	142	282	123	264	289
XR	205	290	254	139	136	282	120	249	321
XR	215	302	263	155	146	286	123	249	321
FX10	208	299	254	113	133	290	120	267	321
FX10	208	299	254	113	133	290	120	267	321
FR	199	299	257	151	123	276	123	244	275
FR	214	299	279	161	133	288	123	244	275

位点 Locus	等位基因数 Alleles	最长片段大小 Max. allele (bp)	最短片段大小 Min. allele (bp)	观测杂合度 Heterozygosity observed (Ho)	期望杂合度 Heterozygosity expected (He)	多态信息含量 Polymorphism information contents (PIC)
SCAAT1	9	252	174	0.3125	0.7601	0.7155
YPL009C	7	305	275	0.4375	0.7944	0.7337
C4	10	303	245	0.4667	0.8598	0.8124
C5	11	172	113	0.2500	0.8669	0.8224
C8	5	146	124	0.2500	0.7802	0.7132
C11	10	290	146	0.4375	0.9032	0.8620
C12	8	159	119	0.5000	0.6956	0.6339
SCAAT3	7	268	243	0.2000	0.8460	0.7934
SCYOR267C	8	321	275	0.3125	0.8206	0.7662

Implantation and Persistence of Inoculated Active Dry Yeast in Industrial Wine Fermentations

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