酿酒酵母糠醛耐受性状相关微卫星标记的筛选

doi:10.13560/j.cnki.biotech.bull.1985.2018-0107

生物技术通报 ›› 2018, Vol. 34 ›› Issue (8): 123-129.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0107

酿酒酵母糠醛耐受性状相关微卫星标记的筛选

李莎莎, 张梁, 石贵阳

1. 粮食发酵工艺与技术国家工程实验室,无锡 214122;
2. 江南大学工业生物技术教育部重点实验室,无锡 214122

收稿日期:2018-01-30 出版日期:2018-08-26 发布日期:2018-09-04
作者简介:李莎莎,女,硕士研究生,研究方向：酿酒酵母的糠醛耐受性;E-mail：442354165@qq.com
基金资助:
江苏省杰出青年基金项目（BK20140002）

Screening of Microsatellite Markers Associated with Furfural Tolerance of Saccharomyces cerevisiae

LI Sha-sha, ZHANG Liang, SHI Gui-yang

1. National Engineering Laboratory for Cereal Fermentation Technology,Jiangsu University,Wuxi 214122;
2. The Key Laboratory of Industrial Biotechnology of Ministry of Education,Jiangnan University,Wuxi 214122

Received:2018-01-30 Published:2018-08-26 Online:2018-09-04

摘要/Abstract

摘要： 筛选两株表型差异单倍体菌株作为亲本,二者杂交获得F₂群体共200株,利用具有多态性的微卫星位点对表型分布在两极端的菌株（39株）进行PCR扩增,运用t检验分析不同位点的基因型在两基因池中的差异显著性,以此获得与糠醛耐受性相关的微卫星位点。结果发现两个与糠醛耐受性有关的微卫星位点,其中2P3位点在两基因池中达到显著差异（P=0.039﹤0.05）,而位点2P5达到极显著差异（P=0.000﹤0.01）;对于微卫星位点2P5,耐受亲本P1在该位点的基因型（333 bp）在子代耐受群体中出现率为72.2%,而敏感亲本P2的基因型（318 bp）在子代敏感群体中出现率达100%（333 bp出现率为零）。上述结果表明微卫星位点2P5的基因型在群体分离中达到极显著差异可能与控制该性状的基因相连锁,在分子设计育种方面可用来选择有利等位基因,提高育种的准确性和预见性。

关键词: 酿酒酵母, 糠醛耐受性, 表型分析, 微卫星标记

Abstract: Two phenotypically different haploid strains were screened as parents and were crossed to obtain two hundred segregants. The polymorphic microsatellite markers were used to amplify offspring yeasts（39 strains）,and t-test was used to analyze the significance of difference among the genotypes of different sites in two gene pools,which is aimed to have microsatellite markers associated with furfural tolerance. The results revealed that two microsatellite markers associated with furfural tolerance were identified. Among them the marker 2P3 had significant differences（P = 0.039 < 0.05）and marker 2P5 had extremely significant differences（P = 0.000 ﹤ 0.01）in the 2 gene pools. Furthermore,72.2% of high tolerant individuals contained the marker 2P5（333 bp）derived from P1,the high tolerant parent;100% of low tolerant individuals contained the marker 2P5（318 bp）derived from P2,the low tolerant parent. These results suggested that the significant difference of 2P5 genotype in population segregation may be linked with the gene controlling this trait and can be used to select the favorable allele in molecular design and breeding so as to improve the accuracy and predictability of breeding.

Key words: Saccharomyces cerevisiae, furfural tolerance, phenotypic analysis, SSR markers

李莎莎, 张梁, 石贵阳. 酿酒酵母糠醛耐受性状相关微卫星标记的筛选[J]. 生物技术通报, 2018, 34(8): 123-129.

LI Sha-sha, ZHANG Liang, SHI Gui-yang. Screening of Microsatellite Markers Associated with Furfural Tolerance of Saccharomyces cerevisiae[J]. Biotechnology Bulletin, 2018, 34(8): 123-129.

参考文献

[1] 张译之, 苟敏, 汤岳琴. 紫外诱变驯化提高酿酒酵母木糖发酵的抑制物耐受性[J]. 生物技术通报, 2017, 33(9):191-199.
[2] Sardi M, Rovinskiy N, Zhang Y, et al.Leveraging genetic-background effects in Saccharomyces cerevisiae to improve lignocellulosic hydrol-ysate tolerance[J]. Applied & Environmental Microbiology, 2016, 82(19):01603-16.
[3] Wang S, Cheng G, Joshua C, et al.Furfural tolerance and detoxification mechanism in Candida tropicalis[J]. Biotechnology for Biofuels, 2016, 9(1):250-261.
[4] Hasunuma T, Ismail KS, et al.Co-expression of TAL1 and ADH1 in recombinant xylose-fermenting Saccharomyces cerevisiae improves ethanol production from lignocellulosic hydrolysates in the presence of furfural[J]. J Biosci Bioeng, 2014, 117(2):165-169.
[5] 赵鲜仙. 酿酒酵母糠醛耐受关键转录调控基因的筛选及其被调控基因的转录分析[D]. 雅安:四川农业大学, 2015.
[6] Putman AI, Carbone I.Challenges in analysis and interpretation of microsatellite data for population genetic studies[J]. Ecology & Evolution, 2014, 4(22):4399-4429.
[7] Tautz D.Hypervariability of simple sequences as a general source for polymorphic DNA markers[J]. Nucleic Acids Research, 1989, 17(16):6463-6471.
[8] 黄龙花, 吴清平, 杨小兵, 等. 基于特定引物PCR的DNA分子标记技术研究进展[J]. 生物技术通报, 2011, 20(2):61-65.
[9] Zhou JC, Yang XL, Cui SL, et al.Correlation between SSR markers and agronomic traits in peanut(Arachis hypogaea L.)[J]. Acta Agronomica Sinica, 2014, 40(7):1197-1205.
[10] Hu XH, Wang MH, Tan T, et al.Genetic dissection of ethanol tolerance in the budding yeast Saccharomyces cerevisiae[J]. Genetics, 2007, 175(3):1479-1487.
[11] Pereira FB, Teixeira MC, Mira NP, et al.Genome-wide screening of Saccharomyces cerevisiae genes required to foster tolerance towards industrial wheat straw hydrolysates[J]. Journal of Industrial Microbiology & Biotechnology, 2014, 41(12):1753-1761.
[12] 李华, 刘丽丽, 李娟. 酿酒酵母产孢培养基的筛选及单倍体的分离[J]. 酿酒科技. 2008, 2(6):22-24.
[13] Huxley C, Green ED, Dunham I.Rapid assessment of S. cerevisiae mating type by PCR[J]. Trends Genet, 1990, 6(8):236-236.
[14] Guldener U, Heck S, Fielder T, et al.A new efficient gene disruption cassette for repeated use in budding yeast[J]. Nucleic Acids Research, 1996, 24(13):2519-2524.
[15] Gietz RD, Schiestl RH, Willems AR, et al.Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure[J]. Yeast, 1995, 11(4):355-360.
[16] Song HS, Jeon JM, Kim HJ, et al.Increase in furfural tolerance by combinatorial overexpression of NAD salvage pathway enzymes in engineered isobutanol-producing E. coli[J]. Bioresource Technology, 2017, 245:1430-1435.
[17] Wu G, Xu Z, Jönsson LJ.Profiling of Saccharomyces cerevisiae transcription factors for engineering the resistance of yeast to lignocellulose-derived inhibitors in biomass conversion.[J]. Microbial Cell Factories, 2017, 16(1):199-214.
[18] Steinmetz LM, Sinha H, Richards DR, et al.Dissecting the architecture of a quantitative trait locus in yeast[J]. Nature, 2002, 416(6878):326-330.
[19] Geng P, et al.Genetic dissection of acetic acid tolerance in Saccha-romyces cerevisiae[J]. World J Microbiol Biotechnol, 2016, 32
(9):1-8.
[20] Ben-Ari G, Zenvirth D, Sherman A, et al.Four linked genes participate in controlling sporulation efficiency in budding yeast[J]. PLoS Genetics, 2006, 2(11):195-205.
[21] Perlstein EO, Ruderfer DM, Roberts DC, et al.Genetic basis of individual differences in the response to small-molecule drugs in yeast[J]. Nature Genetics, 2007, 39(4):496-502.
[22] Swinnen S, Thevelein JM, Nevoigt E.Genetic mapping of quantitative phenotypic traits in Saccharomyces cerevisiae.[J]. Fems Yeast Research, 2012, 12(2):215-227.

酿酒酵母糠醛耐受性状相关微卫星标记的筛选

Screening of Microsatellite Markers Associated with Furfural Tolerance of Saccharomyces cerevisiae

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