石河子不同树龄蟠桃土壤中可培养酵母多样性分析及功能酵母的筛选
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摘要
为了探究新疆本土蟠桃园可培养酵母菌多样性,并挖掘功能酵母资源,本研究以新疆石河子蟠桃园3年、8年、15年树龄的根际和非根际土壤以及桃树叶片为材料,经过传统的分离培养方法获得可培养的酵母菌菌株,并进行形态学、生理生化以及26S r DNA的D1/D2区序列分析,共获得可培养酵母菌129株,从属于12个属17个种,其中子囊菌酵母为优势菌群,占分离属的88%,分布于威克汉姆酵母属(Wickerhamomyces),Vanrija属,Barnettozyma属和有孢圆酵母属(Torulaspora)等11个属的15个种。担子菌占分离属的12%,分布于隐球酵母属(Cryptococcus)的2个种。其中优势属威克汉姆酵母属,包括异常威克汉姆酵母(W. anomalus)和W. pijperi两个种,占总比例的33%,优势种异常威克汉姆酵母所占总株数比例为17%。从可培养酵母中共筛选出23株功能酵母,其中富硒酵母21株,优势种为白地霉(Galactomyces candidum),产蛋白酶酵母2株均属于隐球酵母属的Cryptococcus albidus。结果表明,新疆桃园中蕴含丰富的酵母菌资源,非根际土壤中的酵母多样性大于根际及叶片酵母多样性,且分离得到富硒酵母及产蛋白酶酵母。本研究挖掘了新疆本土可培养酵母菌资源,同时也为功能酵母的开发和利用提供理论指导。
In order to explore the culturable yeast diversity and search for functional yeast resources in Shihezi,Xinjiang,China,yeast strains were obtained by traditional isolation and culture methods from the rhizosphere and non-rhizosphere soils and the flat peach leaves of the 3,8 and 15 years. Using the morphological,physiological and biochemical methods and the sequence analysis of D1/D2 region of 26 S r DNA,we obtained a total of 129 strains of culturable yeast,belonging to 12 genera and 17 species,of which ascomycete yeast was the dominant community,accounting for 88% of the isolated genera and distributed in 15 species of 11 genera. Basidiomycetes accounted for 12% of the isolated genera and distributed in two species of Cryptococcus. Among them,Wickerhamomyces,including anomalus and pijperi,accounted for 33% of the total,and the dominant species W. anomalu accounted for 17% of the total number of yeasts. A total of 23 functional yeasts were screened,including 21 selenium-enriched yeasts,the dominant species was Galactomyces candidum,and the two protease-producing yeasts belonged to C. albidus. The results showed that abundant yeast resources were contained in peach tree soil in Xinjiang. The yeast diversity in non-rhizosphere soil was greater than that in rhizosphere soil and leaves,and selenium-enriched yeast and protease-producing yeast were also isolated. The study explored the culturable yeast resources in Xinjiang and provides theoretical guidance for the development and utilization of functional yeast.
In order to explore the culturable yeast diversity and search for functional yeast resources in Shihezi,Xinjiang,China,yeast strains were obtained by traditional isolation and culture methods from the rhizosphere and non-rhizosphere soils and the flat peach leaves of the 3,8 and 15 years. Using the morphological,physiological and biochemical methods and the sequence analysis of D1/D2 region of 26 S r DNA,we obtained a total of 129 strains of culturable yeast,belonging to 12 genera and 17 species,of which ascomycete yeast was the dominant community,accounting for 88% of the isolated genera and distributed in 15 species of 11 genera. Basidiomycetes accounted for 12% of the isolated genera and distributed in two species of Cryptococcus. Among them,Wickerhamomyces,including anomalus and pijperi,accounted for 33% of the total,and the dominant species W. anomalu accounted for 17% of the total number of yeasts. A total of 23 functional yeasts were screened,including 21 selenium-enriched yeasts,the dominant species was Galactomyces candidum,and the two protease-producing yeasts belonged to C. albidus. The results showed that abundant yeast resources were contained in peach tree soil in Xinjiang. The yeast diversity in non-rhizosphere soil was greater than that in rhizosphere soil and leaves,and selenium-enriched yeast and protease-producing yeast were also isolated. The study explored the culturable yeast resources in Xinjiang and provides theoretical guidance for the development and utilization of functional yeast.
引文
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[24]Singhal A,Jaiswal P K,Jha P K,et al.Assesment of Cryptococcus albidus for biopulping of eucalyptus[J].Prep Biochem Biotech,2013,43(8):735-749.
[2]Tan J L,Li Z Y,Zhou B,et al.Extracellular enzymatic activities of yeasts isolated from Fuxian Lake and Xingyun Lake in Yunnan Plateau[J].Microbiology China,2018,45(2):302-313,333.谭金连,李治滢,周斌,等.云南高原湖泊抚仙湖和星云湖的酵母菌胞外酶活性[J].微生物学通报,2018,45(2):302-313,333.
[3]Feng T,Wang X Z,Wang Y F,et al.Isolation and identification of yeast strains from vineyard soils and aroma compounds of wines fermented by them[J].Food Science,2018,39(14):213-220.冯涛,王旭增,王一非,等.葡萄园土壤中酿酒酵母的分离鉴定及其发酵葡萄酒香气成分分析[J].食品科学,2018,(14):213-220.
[4]Zhao X N,Wang H X,Luo F,et al.Isolation and identification of yeast strains on wine grapes in Hebei region[J].Sino-Overseas Grapevine&Wine,2018(1):14-19.赵晓宁,王焕香,罗飞,等.河北产区酿酒葡萄酵母菌的分析及鉴定[J].中外葡萄与葡萄酒,2018(1):14-19.
[5]Gao B Y,Chen X K,Xu B,et al.Screening of yeast strains producing secretory proteinase[J].Journal of Fudan University(Natural Science),1999,38(5):537-539.高卜渝,陈孝康,徐波,等.产分泌蛋白酶酵母菌株的筛选[J].复旦学报:自然科学版,1999,38(5):537-539.
[6]Lou X D,Zhang G L,Ye B C.Screening of high selenium-enriched yeast and enhancement of its selenium-enriched capacity[J].Biotechnology Bulletin,2017,33(12):132-137.娄兴丹,张根林,叶邦策.高富硒酵母的筛选及富硒强化研究[J].生物技术通报,2017,33(12):132-137.
[7]Kurtzman C P,Fell J W,Boekhout T.2011.The yeasts:a taxonomic study[M].5th ed.Amsterdam:Elsevier Science.2011
[8]Kurtzman C P,Fell J W,Boekhout T,et al.Methods for isolation,phenotypic characterization and maintenance of yeasts//The yeasts:a taxonomic study[M].5th ed.Amsterdam:Elsevier Science.2011.87-110
[9]Lu W J.Diversity analysis of yeast from raw milk and cheese in Tacheng District of Xinjiang[D].Shihezi:Shihezi University,2016.芦文娟.新疆塔城地区原料乳及奶酪中酵母菌多样性的研究[D].石河子:石河子大学,2016.
[10]Caba?es F J,Vega S,CastelláG.Malassezia cuniculi sp.nov.,a novel yeast species isolated from rabbit skin[J].Med Mycol,2011,49(1):40-48.
[11]Lou Q Z,Lu M,Sun J H.Yeast diversity associated with invasive Dendroctonus valens killing Pinus tabuliformis in China using culturing and molecular methods[J].Microb Ecol,2014,68(2):397-415.
[12]Zhao Y T,Sun H L,Liu M R,et al.Screening and phylogenetic analysis of yeast producing protease from permafrost of the Glacier No.1 in the Tianshan Mountains[J].Journal of Anhui Agriculture Science,2015,43(1):248-250.赵有婷,孙海龙,刘敏瑞,等.天山一号冰川产蛋白酶酵母菌的筛选及其系统发育分析[J].安徽农业学,2015,43(1):248-250.
[13]Solis M J L,Yurkov A,Dela Cruz T E,et al.Leaf-inhabiting endophytic yeasts are abundant but unevenly distributed in three Ficus species from botanical garden greenhouses in Germany[J].Mycol Prog,2015,14(1):1019.
[14]Shao B J,Tao Y Q,Wang S Y,et al.Isolation,identification,and screening of dominant yeast strains in fried cake made from natural fermented glutinous rice[J].Modern Food Science and Technology,2016,32(7):91-96.邵冰洁,陶永清,王素英,等.传统法糯米发酵炸糕中酵母菌的分离鉴定及优势菌株筛选[J].现代食品科技,2016,32(7):91-96.
[15]Izawa N,Kudo M,Nakamura Y,et al.Production of aroma compounds from whey using Wickerhamomyces pijperi[J].Amb Express,2015,5(1):1-9.
[16]Wang H.The diversity and edible safety of yeasts on the surface of apple and pear[D].Jinan:Qilu University of Technology,2013.王红.苹果和梨果表面酵母多样性及其食用安全性研究[D].济南:齐鲁工业大学,2013.
[17]Kaur P,Kunze G,Satyanarayana T.Yeast phytases:present scenario and future perspectives[J].Crit Rev Biotechnol,2007,27(2):93-109.
[18]Alves-Araújo C,Pacheco A,Almeida M J,et al.Sugar utilization patterns and respiro-fermentative metabolism in the baker.s yeast Torulaspora delbrueckii[J].Microbiology,2007,153(3):898-904.
[19]Morais C G,Cadete R M,Uetanabaro A P,et al.D-xylose-fermenting and xylanase-producing yeast species from rotting wood of two Atlantic Rainforest habitats in Brazil[J].Fungal Genet Biol,2013,60(6):19-28.
[20]Kennedy T A,Naeem S,Howe K M,et al.Biodiversity as a barrier to ecological invasion[J].Nature,2002,417(6889):636-638.
[21]Wang H D,Zhang L L,Zhu Z H.Effects of clipping and fertilizing on the relationships between species diversity and ecosystem functioning and mechanisms of community stability in alpine meadow[J].Chinese Journal of Plant Ecology,2013,37(4):279-295.王海东,张璐璐,朱志红.刈割、施肥对高寒草甸物种多样性与生态系统功能关系的影响及群落稳定性机制[J].植物生态学报,2013,37(4):279-295.
[22]Zhu J,Shi J L,Liu L P.GC/MS analysis of the aroma compounds produced in pear pomace by Geotrichum fungi[J].Journal of Northwest A&F University(Nat.Sci.Ed.),2010,38(2):171-178.朱静,师俊玲,刘拉平.地霉属菌株发酵梨渣产香气成分的GC/MS分析[J].西北农林科技大学学报(自然科学版),2010,38(2):171-178.
[23]Sun B S,Zheng L L,Liu T M,et al.Applied research progress in Geotrichum candidum[J].Food Research and Development,2008,29(9):163-166.孙丙升,郑莉莉,刘天明,等.白地霉的应用研究进展[J].食品研究与开发,2008,29(9):163-166.
[24]Singhal A,Jaiswal P K,Jha P K,et al.Assesment of Cryptococcus albidus for biopulping of eucalyptus[J].Prep Biochem Biotech,2013,43(8):735-749.