云南不同仓储地复烤烟叶表面醇化期间可培养真菌的动态变化
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摘要
为了解云南复烤烟叶(K326)异地醇化过程可培养真菌的多样性,通过形态特征观察、18S rDNA序列分析研究了异地醇化过程中真菌菌群的结构。结果表明,从220份复烤烟叶样品中共分离、纯化到2 963株可培养真菌。4个醇化地烟叶表面真菌数量随醇化时间的延长呈"下降-上升-下降-趋于平缓"的趋势。结合形态特征和18S rDNA序列的分析结果,共鉴定出39个属(种)的真菌。不同醇化地、不同醇化时间复烤烟叶表面真菌的种群结构及其变化规律均有差异,但根霉属真菌始终为优势菌群。
To investigate the diversity of culturable fungi on the surface of redried tobacco leaves of K326 during aging in different areas of Yunnan, the morphological features and 18 S rDNA sequences of the fungi were analyzed to study the community structure. The results showed that a total of 2 963 culturable fungi were obtained from 220 samples of redried tobacco leaves via separation and purification. With prolonged aging time,the quantity of culturable fungi on the surface of tobacco leaves from four areas presented a down-up-downstable trend. By morphological features and 18 S rDNA sequences, 39 genera(species) were identified. The community structure and variation of fungi differed on the surface of redried tobacco leaves according to different aging times and areas. However, the dominant genus was Rhizopus spp.
To investigate the diversity of culturable fungi on the surface of redried tobacco leaves of K326 during aging in different areas of Yunnan, the morphological features and 18 S rDNA sequences of the fungi were analyzed to study the community structure. The results showed that a total of 2 963 culturable fungi were obtained from 220 samples of redried tobacco leaves via separation and purification. With prolonged aging time,the quantity of culturable fungi on the surface of tobacco leaves from four areas presented a down-up-downstable trend. By morphological features and 18 S rDNA sequences, 39 genera(species) were identified. The community structure and variation of fungi differed on the surface of redried tobacco leaves according to different aging times and areas. However, the dominant genus was Rhizopus spp.
引文
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[28]苏加坤,徐达,郭磊,等.基于宏基因组测序的烟叶表面微生物多样性分析[J].基因组学与应用生物学,2017,36(4):1538-1545.SU Jiakun,XU Da,GUO Lei,et al.Diversity analysis of microorganism on flue-cured tobacco leaf surface based on metagenome sequencing[J].Genomics and Applied Biology,2017,36(4):1538-1545.
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[31]徐洁.两个真菌菌株对烤烟诱香机理的初步研究[D].泰安:山东农业大学,2006.XU Jie.A preliminary study on mechanisms of aromaproducing in flue-cured tobacco induced by two fungal isolates[D].Tai’an:Shandong Agricultural University,2006.
[2]孙敬国,闫铁军,司辉,等.烤烟醇化过程中酶的作用与变化特征研究进展[J].湖北农业科学,2008,47(10):1207-1211.SUN Jingguo,YAN Tiejun,SI Hui,et al.Advances in enzyme characterizations of flue-cured tobacco during the aging process[J].Hubei Agricultural Sciences,2008,47(10):1207-1211.
[3]朱大恒,陈锐,陈再根,等.烤烟自然醇化与人工发酵过程中微生物变化及其与酶活性关系的研究[J].中国烟草学报,2001,7(2):26-30.ZHU Daheng,CHEN Rui,CHEN Zaigen,et al.The relationship between microorgnisms and enzyme activities in flue-cured tobacco during aging and fermentation[J].Acta Tabacaria Sinica,2001,7(2):26-30.
[4]王志愿,姜清治,霍沁建.生物技术在烟草品质改良中的应用研究[J].贵州农业科学,2011,39(1):16-20.WANG Zhiyuan,JIANG Qingzhi,HUO Qinjian.Research progress on application of biotechnology in quality improvement of tobacco[J].Guizhou Agricultural Sciences,2011,39(1):16-20.
[5]夏露,张娟,王远亮,等.生物技术在烟叶发酵中的应用研究进展[J].安徽农业科学2010,38(22):12013-12015.XIA Lu,ZHANG Juan,WANG Yuanliang,et al.Research progress on the application of biotechnology in tobacco fermentation[J].Journal of Anhui Agricultural Sciences,2010,38(22):12013-12015.
[6]谢和,韩忠礼,赵维娜.微生物发酵对烤烟内在品质的影响[J].山地农业生物学报,1999,18(4):227-230.XIE He,HAN Zhongli,ZHAO Weina.Effect of microbial fermentation on internal quality of flue-cured tobacco[J].Journal of Mountain Agriculture and Biology,1999,18(4):227-230.
[7]唐丽,周冰,华一崑,等.醇化技术在改善烟叶品质中的研究进展[J].江西农业学报,2013,25(7):53-56.TANG Li,ZHOU Bing,HUA Yikun,et al.Research advance in alcoholization technology for improving quality of tobacco leaf[J].Acta Agriculturae Jiangxi,2013,25(7):53-56.
[8]李若铭.烟草微生物应用于烟叶发酵的研究进展[J].科技创新导报,2010(35):4,6.LI Ruoming.The microbial fermentation applied research progress of tobacco[J].Science and Technology Innovation Herald,2010(35):4,6.
[9]张成省,林建胜,孔凡玉,等.山东仓储片烟表面微生物区系研究[J].中国烟草学报,2010,16(4):58-62,66.ZHANG Chengsheng,LIN Jiansheng,KONG Fanyu,et al.Research on phyllosphere microflora in stored fluecured tobacco strips in Shandong Province[J].Acta Tabacaria Sinica,2010,16(4):58-62,66.
[10]Wang J J,Xu Z C,Fan J L,et al.Effects of X-ray irradiation on the microbial growth and quality of fluecured tobacco during aging[J].Radiation Physics and Chemistry,2015,111:9-13.
[11]贺升华,任炜.烤烟气象[M].昆明:云南科技出版社,2001.HE Shenghua,REN Wei.Flue-cured tobacco meteorology[M].Kunming:Yunnan Science&Technology Press,2001.
[12]中国农业科学院烟草研究所.中国烟草栽培学[M].上海:上海科学技术出版社,2005.Institute of Tobacco Research of CAAS.Tobacco cultivation in China[M].Shanghai:Shanghai Scientific and Technical Publishers,2005.
[13]张谦,胡剑波.新形势下云南烟草经济发展的SWOT分析与对策[J].安徽农业科学,2011,39(6):3722-3725.ZHANG Qian,HU Jianbo.SWOT analysis and countermeasures of economic development of tobacco in Yunnan province under the new situation[J].Journal of Anhui Agricultural Sciences,2011,39(6):3722-3725.
[14]胡剑波,杨虹.WHO FCTC对云南烟草经济的负面影响与对策探析[J].云南财经大学学报,2010,26(2):134-139.HU Jianbo,YANG Hong.An analysis of the negative influence of WHO FCTC on the tobacco economy of Yunnan and the countermeasures[J].Journal of Yunnan University of Finance&Economics,2010,26(2):134-139.
[15]张亚恒,王芳,张庆明,等.微生物在烟叶醇化和再造烟叶生产中的应用进展[J].湖北农业科学,2015,54(18):4390-4393.ZHANG Yaheng,WANG Fang,ZHANG Qingming,et al.Application of microbiology in flue-cured tobacco leaves during aging and reconstituted tobacco production[J].Hubei Agricultural Sciences,2015,54(18):4390-4393.
[16]Ellis M B.Dematiaceous hyphomycetes[M].Egham:International Mycological Institute,1988.
[17]Barnett H L,Hunter B B.Illustrated genera of imperfect fungi[M].New York:Macmillan Publishing Company,1987.
[18]Sutton B C.The Coelomycetes.Fungi imperfecti with pycnidia,acervuli and stromata[M].Kew:Commonwealth Mycological Institute,1980.
[19]冯智红,严培兰,王建华,等.香菇中污染真菌的分离与鉴定[J].上海农业学报,2015,31(4):50-53.FENG Zhihong,YAN Peilan,WANG Jianhua,et al.Isolation and identification of fungus contaminated in Lentinula edodes[J].Acta Agriculturae Shanghai,2015,31(4):50-53.
[20]燕勇,李卫平,高雯洁,等.rDNA-ITS序列分析在真菌鉴定中的应用[J].中国卫生检验杂志,2008,18(10):1958-1961.YAN Yong,LI Weiping,GAO Wenjie,et al.Application of rDNA ITS sequence analysis in fungus identification[J].Chinese Journal of Health Laboratory Technology,2008,18(10):1958-1961.
[21]Hibbett D,Abarenkov K,K?ljalg U,et al.Sequencebased classification and identification of Fungi[J].Mycologia,2016,108(6):1049-1068.
[22]K?ljalg U,Nilsson R H,Abarenkov K,et al.Towards a unified paradigm for sequence-based identification of fungi[J].Molecular Ecology,2013,22(21):5271-5277.
[23]浦绍占.不同仓储条件对醇化烟叶品质的影响[D].昆明:昆明理工大学,2017.PU Shaozhan.The effect of different warehouse conditions on quality of aged tobacco[D].Kunming:Kunming University of Science and Technology,2017.
[24]赵铭钦,邱立友,张维群,等.陈化期间烤烟叶片中生物活性变化的研究[J].华中农业大学学报,2000,19(6):537-542.ZHAO Mingqin,QIU Liyou,ZHANG Weiqun,et al.Study on the changes of biological activity of fluecured tobacco leaves during aging[J].Journal of Huazhong Agricultural University,2000,19(6):537-542.
[25]单宏英.陈化烟叶表面有益微生物的分离筛选、鉴定及应用研究[D].杨凌:西北农林科技大学,2012.SHAN Hongying.Studies on screening identification and application of beneficial microorganisms isolated from aging tobacco surfaces[D].Yangling:Northwest A&F University,2012.
[26]汪长国,李宁,寇明钰,等.复烤烟叶异地醇化过程中生物活性的变化[J].中国农业大学学报,2013,18(2):105-109.WANG Changguo,LI Ning,KOU Mingyu,et al.Changes of biological activity of flue-cured tobacco leaves from different places aging[J].Journal of China Agricultural University,2013,18(2):105-109.
[27]龚俊.烤后片烟储存过程中微生物多样性及变化动态[D].上海:华东师范大学,2015.GONG Jun.The diversity and dynamic of microorganism on flue-cured tobacco leaves during different aged phases[D].Shanghai:East China Normal University,2015.
[28]苏加坤,徐达,郭磊,等.基于宏基因组测序的烟叶表面微生物多样性分析[J].基因组学与应用生物学,2017,36(4):1538-1545.SU Jiakun,XU Da,GUO Lei,et al.Diversity analysis of microorganism on flue-cured tobacco leaf surface based on metagenome sequencing[J].Genomics and Applied Biology,2017,36(4):1538-1545.
[29]邱立友,赵铭钦,岳雪梅,等.自然发酵烤烟叶面微生物区系的分离鉴定[J].烟草科技,2000(3):14-17.QIU Liyou,ZHAO Mingqin,YUE Xuemei,et al.Isolation and identification of the microflora on tobacco leaves during the natural fermentation of flue-cured tobacco[J].Tobacco Science&Technology,2000(3):14-17.
[30]Welty R E,Lucas G B.Fungi isolated from damaged flue-cured tobacco[J].Applied Microbiology,1968,16(6):851-854.
[31]徐洁.两个真菌菌株对烤烟诱香机理的初步研究[D].泰安:山东农业大学,2006.XU Jie.A preliminary study on mechanisms of aromaproducing in flue-cured tobacco induced by two fungal isolates[D].Tai’an:Shandong Agricultural University,2006.