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Identification method for sulfur-resistant hybrid breeding progeny of saccharomyces bayanus

An identification method and hybrid breeding technology, which are applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of low accuracy, inability to distinguish hybrid offspring from hybrid parents, and inability to observe differences, so as to improve accuracy. rate effect

Inactive Publication Date: 2014-04-23
SOUTHWEST FORESTRY UNIVERSITY
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Problems solved by technology

The microscopic morphology of Saccharomyces species mainly exists in the form of single cells. Microscopic observation results show that the cell properties of various strains are very close, the accuracy rate is low, and almost no difference can be observed. Parents are separated

Method used

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  • Identification method for sulfur-resistant hybrid breeding progeny of saccharomyces bayanus
  • Identification method for sulfur-resistant hybrid breeding progeny of saccharomyces bayanus
  • Identification method for sulfur-resistant hybrid breeding progeny of saccharomyces bayanus

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Embodiment Construction

[0051] Materials: non-sulfur-resistant yeast strains are 7A2, 7A6, 7D4, and sulfur-resistant yeast strains are A9 strains (both quoted from the University of Auckland, New Zealand, introduced by: Zhang Hanyao, 2010), all kept in this laboratory, or can be sent to Upon request, Professor Richard Gardner, University of Auckland. Contact: Wine Science Programme, School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand.

[0052] Reagents: primers were synthesized by Shanghai Bioengineering Company; Taq DNA polymerase, dNTPs, and RNase were purchased from Shanghai Bioengineering Company; DNA Marker was 1Kb plus ladder, purchased from Invitrogen; Yeast Genomic DNA Extraction Kit was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd. Other conventional reagents were domestic analytically pure products .

[0053] (1) Breeding Sulfur-resistant Saccharomyces japonicus strains and DNA sequence identification

[0054] Bay Yeas...

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Abstract

The invention discloses an identification method for a sulfur-resistant hybrid breeding progeny of saccharomyces bayanus, belonging to an identification method for a novel saccharomyces bayanus breeding strain. The identification method comprises the following steps: saccharomyces bayanus hybridization; sulfur resistance measurement of a saccharomyces bayanus strain; saccharomyces bayanus DNA (Deoxyribonucleic Acid) extraction and purification, wherein a PCR (Polymerase Chain Reaction) system taking (i) FZF1 ( / i) as a primer and a procedure are adopted; PCR primer sequencing and identifying; and further ISSR (Inter-Simple Sequence Repeat) analysis for the sulfur-resistant hybrid breeding progeny. In the embodiment, 18 suspected hybrid progenies are obtained from 3 hybridized combinations. The PCR analysis results are as follows: the 18 suspected hybrid progenies contain 5 strains only containing sulfur-nonresistant parental characteristic bands, 2 strains containing sulfur-resistant parental characteristic bands and other 11 strains containing biparental characteristic bands. The ISSR analysis for 8 sulfur-resistant hybrid progenies proves that the same strain has two single colonies with the completely-consistent fingerprint spectrums. According to the invention, 6 sulfur-resistant hybrid breeding progeny strains of saccharomyces bayanus are obtained according to a DNA sequence and an ISSR atlas and through sulfur-resistant phenotype screening, so that the accuracy rate for identifying the hybrid breeding progeny strains is remarkably increased.

Description

technical field [0001] The invention belongs to yeast breeding and identification methods. Background technique [0002] Yeast selection is one of the core technologies of high-quality fruit wine brewing, which has a decisive impact on the aroma and quality of fruit wine. At present, the yeasts used in the brewing of fruit wine in our country are basically imported, and the homogeneity of the quality of fruit wine is becoming more and more serious, which restricts the development and competitiveness of new quality fruit wine in our country, and also threatens the wild yeast resources in our country. Saccharomyces cerevisiae( Saccharomycess cerevisiae ) is the yeast commonly used in various types of wine production, but the wine brewed with Saccharomyces cerevisiae as a fermentation strain has a flat flavor, while the generalized Saccharomyces cerevisiae group ( Saccharomycesses lato) has been extended to the breeding of other yeasts, such as S. Saccharomyces bayanus )...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156C12Q2600/172
Inventor 刘小珍张汉尧
Owner SOUTHWEST FORESTRY UNIVERSITY
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