Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (9): 2006-2016.doi: 10.3864/j.issn.0578-1752.2021.09.016

• FOOD SCIENCE AND ENGINEERING • Previous Articles     Next Articles

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

SUN Yue1(),YANG HuiMin2,HE RongRong1,ZHANG JunXiang1()   

  1. 1School of Food and Wine, Ningxia University, Yinchuan 750021
    2College of Enology, Northwest A&F University, Yangling 712100, Shaanxi
  • Received:2020-08-24 Accepted:2020-11-30 Online:2021-05-01 Published:2021-05-10
  • Contact: JunXiang ZHANG E-mail:yuesun86@126.com;zhangjunxiang@126.com

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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

Table 1

Information of SSR markers used in this study"

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]

Fig. 1

Interdelta fingerprinting patterns of four ADY FR1, FR2 and XR1, XR2 represent different Interdelta patterns of the corresponding ADY"

Table 2

SSR results of four ADY"

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

Fig. 2

Interdelta fingerprinting patterns of S. cerevisiae isolated from industrial wine fermentations of BDX, XR, FR and FX10"

Table 3

Genetic diversity of S. cerevisiae strains isolated from Cabernet Sauvignon industrial wine fermentation"

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

Fig. 3

Dynamics changes of different S. cerevisiae strains during inoculated fermentation of BDX, XR, FR and FX10 A: Inoculated with BDX; B: Inoculated with XR; C: Inoculated with FR; D: Inoculated with FX10"

Fig. 4

UPGMA dendrograms showing the genetic relationships between ADY and Ningxia indigenous S. cerevisiae strain"

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