Fungal composition and diversity analysis of healthy and rotten tobacco leaves after curing

doi:10.3969/j.issn.1004-1524.2020.06.10

›› 2020, Vol. 32 ›› Issue (6): 1019-1028.DOI: 10.3969/j.issn.1004-1524.2020.06.10

• Plant Protection • Previous Articles Next Articles

Fungal composition and diversity analysis of healthy and rotten tobacco leaves after curing

CHEN Qianli^1,2, WANG Hancheng², LIANG Yongjin^{3, *}, CAI Liuti², HUANG Yu^1,2, ZHOU Hao⁴, LI Zhong¹, HAN Jie⁵

1. College of Agriculture, Guizhou University, Guiyang 550025, China;
2. Guizhou Academy of Tobacco Science, Guiyang 550081, China;
3. Guangxi Zhongyan Industry Co. Ltd., Nanning 530001, China;
4. College of Life Sciences, Yangtze University, Jingzhou 434025, China;
5. College of Basic Medical Sciences, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China

Received:2019-10-10 Online:2020-06-25 Published:2020-06-24

Abstract

Abstract: To explore difference of the fungal community composition of healthy and rotten tobacco leaves after curing, the fungal community composition of both healthy and tobacco pole rot samples from flue-cured tobacco leaves were investigated by using PCR amplification of the ITS1-ITS2 region. The amplified fragments were sequenced using Illumina Miseq high-throughput sequencing technique. It was shown that the fungi from healthy tobacco (MJ), rotten tobacco (MB) were distributed in 6 phyla (Ascomycota, Basidiomycota, Zygomycota, etc), 25 classes, 58 orders, 187 families, 260 genera and 394 fungal taxa (OTUs). The dominant genus of MJ was Alternaria, followed by Aspergillus, Neocamarosporium, Gibberella and Acremonium. For MB, the dominant genus was Aspergillus, followed by Penicillium and Rhizopus. Alpha index showed that the fungal community richness and diversity of MJ were higher than those of MB. Additionally, functional assignments were made, the results showed that 71.06% of total OTUs belonged to eight trophic modes, of which saprotroph was the main group. The results of this study revealed the community structure of healthy and rotten tobacco leaves after curing and provided reference basis for formulating the corresponding control plan.

Key words: rotten tobacco leaf, fungal ITS area, community structure, Illumina high throughput sequencing

CLC Number:

S572

CHEN Qianli, WANG Hancheng, LIANG Yongjin, CAI Liuti, HUANG Yu, ZHOU Hao, LI Zhong, HAN Jie. Fungal composition and diversity analysis of healthy and rotten tobacco leaves after curing[J]. , 2020, 32(6): 1019-1028.

References

[1] BURTON H R, ANDERSEN R A, FLEMING P D, et al.Changes in chemical composition of burley tobacco during sensecence and curing. 2. Acylated pyridine alkaloids[J]. Journal of Agricultural and Food Chemistry, 1988, 36(3): 579-584.
[2] MORIN A, PORTER A, JOLY J, et al.Evolution of tobacco-specific nitrosamines and microbial populations during flue-curing of tobacco under direct and indirect heating[J]. Beiträge Zur Tabakforschung International, 2004, 21(1): 40-46.
[3] KORTEKAMP A, SCHMIDTKE M, SERR A.Infection and decay of tobacco caused by Rhizopus oryzae[J]. Journal of Plant Diseases and Protection, 2003, 110:535-543.
[4] SCHLOTZHAUER W S, HIGMAN E B, SCHMELTZ I.Pyrolysis of tobacco extracts[M]//The chemistry of tobacco and tobacco smoke. Boston, MA: Springer US, 1972: 65-76.
[5] 陈二龙, 苏家恩, 范志勇, 等. 烘烤过程中霉变烟叶主脉超微结构的变化[J]. 湖南农业大学学报(自然科学版), 2017, 43(6): 626-629.
CHEN E L, SU J E, FAN Z Y, et al.Changes of ultrastructure of midrib in mildew tobacco leaves during flue curing[J]. Journal of Hunan Agricultural University(Natural Sciences), 2017, 43(6): 626-629.(in Chinese with English abstract)
[6] 彭清云, 易图永. 烟叶霉变的原因及其防治研究进展[J]. 中国农学通报, 2007, 23(11): 146-150.
PENG Q Y, YI T Y.Analysis of tobacco mildew and its control measures[J]. Chinese Agricultural Science Bulletin, 2007, 23(11): 146-150.(in Chinese with English abstract)
[7] 顾钢, 肖顺, 周挺, 等. 烘烤期烟叶霉烂病的研究进展与展望[J]. 中国烟草学报, 2018, 24(5): 112-116.
GU G, XIAO S, ZHOU T, et al.Development and prospect of research in tobacco leaf mildew rot during flue-curing[J]. Acta Tabacaria Sinica, 2018, 24(5): 112-116.(in Chinese with English abstract)
[8] 苏家恩, 典瑞丽, 陈二龙, 等. 烘烤过程中烟叶霉变研究进展[J]. 安徽农业科学, 2018, 46(19): 16-18.
SU J E, DIAN R L, CHEN E L, et al.Research advances in tobacco leaf mold during baking[J]. Journal of Anhui Agricultural Sciences, 2018, 46(19): 16-18.(in Chinese with English abstract)
[9] SCHWARTZ K, SHERLOCK G. High-throughput yeast strain sequencing[J]. Cold Spring Harbor Protocols, 2016, 2016(10): pdb.top077651.
[10] 秦楠, 栗东芳, 杨瑞馥. 高通量测序技术及其在微生物学研究中的应用[J]. 微生物学报, 2011, 51(4): 445-457.
QIN N, LI D F, YANG R F.Next-generation sequencing technologies and the application in microbiology: a review[J]. Acta Microbiologica Sinica, 2011, 51(4): 445-457.(in Chinese with English abstract)
[11] DONG L L, XU J, ZHANG L J, et al.High-throughput sequencing technology reveals that continuous cropping of American ginseng results in changes in the microbial community in arable soil[J]. Chinese Medicine, 2017, 12: 18.
[12] LI Z Y, LIU X, LIU R R, et al.Insight into bacterial community diversity and monthly fluctuations of Medicago sativa rhizosphere soil in response to hydrogen gas using illumina high-throughput sequencing[J]. Current Microbiology, 2018, 75(12): 1626-1633.
[13] 刘强, 赵本宣, 李汝伟, 等. 舟山群岛不同功能区划海域细菌群落结构分析[J]. 微生物学报, 2018, 58(2): 247-263.
LIU Q, ZHAO B X, LI R W, et al.Bacterial community structure in different functional sea areas of Zhoushan archipelago[J]. Acta Microbiologica Sinica, 2018, 58(2): 247-263.(in Chinese with English abstract)
[14] 白洁, 刘小沙, 侯瑞, 等. 南海南部海域浮游细菌群落特征及影响因素研究[J]. 中国环境科学, 2014, 34(11): 2950-2957.
BAI J, LIU X S, HOU R, et al.Community structure and influencing factors of bacterioplankton in the southern South China sea[J]. China Environmental Science, 2014, 34(11): 2950-2957.(in Chinese with English abstract)
[15] 范忠原, 刘晗璐, 王峰, 等. 基于高通量测序技术研究水貂远端肠道细菌群落组成[J]. 微生物学通报, 2016, 43(1): 123-130.
FAN Z Y, LIU H L, WANG F, et al.Determination of bacterial community composition in the distal gut of mink (Mustela vison) by high-throughput sequencing[J]. Microbiology China, 2016, 43(1): 123-130. (in Chinese with English abstract)
[16] ISHII S, HOSHINO B, NURTAZIN S T.Properties and seasonal variation of milk produced by one-humped camels (Camelus dromedarius) and two-humped camels (C. bactrianus) in the Republic of Kazakhstan[J]. Journal of Arid Land Studies, 2017, 26(6): 219-222.
[17] JU F, ZHANG T.Novel microbial populations in ambient and mesophilic biogas-producing and phenol-degrading consortia unraveled by high-throughput sequencing[J]. Microbial Ecology, 2014, 68(2): 235-246.
[18] 张昊. 典型城市空气颗粒物中微生物的群落结构及环境因素影响研究[D]. 杭州: 浙江大学, 2018.
ZHANG H.Study of airborne microbial community structure and environmental factors in typical urban airborne particles[D]. Hangzhou: Zhejiang University, 2018.(in Chinese with English abstract)
[19] 常安然, 李佳, 张耸, 等. 基于宏基因组学16S rDNA测序对烟草根际土壤细菌群落组成分析[J]. 中国农业科技导报, 2017, 19(2): 43-50.
CHANG A R, LI J, ZHANG S, et al.Analysis of bacterial community structure in rhizosphere soil of tobacco based on the metagenomics 16S rDNA sequencing technology[J]. Journal of Agricultural Science and Technology, 2017, 19(2): 43-50.(in Chinese with English abstract)
[20] 李珊辉. 卷烟加工及储存过程中微生物群落结构与其品质关系的研究[D]. 昆明: 云南大学, 2015.
LI S H.Study on the relation between microbial community structure and the quality in cigarette processing and storage process[D]. Kunming: Yunnan University, 2015.(in Chinese with English abstract)
[21] 龚俊, 刘玉配, 李媛媛. 烟叶表面微生物类群两种检测方法的比较研究[J]. 华东师范大学学报(自然科学版), 2016(3): 92-101.
GONG J, LIU Y P, LI Y Y.Comparative analysis of microbial communities on tobacco leaves between clone library and high-throughput sequencing[J]. Journal of East China Normal University(Natural Science), 2016(3): 92-101.(in Chinese with English abstract)
[22] 李秀妮, 李猛, 万德建, 等. 烟叶微生物及其在烟叶发酵和醇化中的作用研究进展[J]. 微生物学通报, 2019, 46(6): 1520-1529.
LI X N, LI M, WAN D J, et al.Role of microorganisms in tobacco fermentation and alcoholization: a review[J]. Microbiology China, 2019, 46(6): 1520-1529.(in Chinese with English abstract)
[23] 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 J K, XU D, GUO L, 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.(in Chinese with English abstract)
[24] 刘开朗, 王加启, 卜登攀, 等. 环境微生物群落结构与功能多样性研究方法[J]. 生态学报, 2010, 30(4): 1074-1080.
LIU K L, WANG J Q, BU D P, et al.Current progress in approaches to the study of structure and function diversities of environmental microbial communities[J]. Acta Ecologica Sinica, 2010, 30(4): 1074-1080.(in Chinese with English abstract)
[25] ANGELINI P, RUBINI A, GIGANTE D, et al.The endophytic fungal communities associated with the leaves and roots of the common reed (Phragmites australis) in Lake Trasimeno (Perugia, Italy) in declining and healthy stands[J]. Fungal Ecology, 2012, 5(6): 683-693.
[26] 陈善义, 李菁菁, 林俭, 等. 基于高通量测序的不同产地和部位自然醇化片烟真菌群落结构分析[J]. 烟草科技, 2018, 51(4): 12-19.
CHEN S Y, LI J J, LIN J, et al.High-throughput sequencing fungal community structures in aging tobacco strips from different growing areas and stalk positions[J]. Tobacco Science & Technology, 2018, 51(4): 12-19.(in Chinese with English abstract)
[27] 陈乾丽, 李忠, 汪汉成, 等. 烤后不同霉变程度烟叶际真菌群落组成与多样性分析[J]. 微生物学报, 2019, 59(12): 2401-2409.
CHEN Q L, LI Z, WANG H C, et al.Fungal composition and diversity of tobacco phyllosphere from cured tobacco leaves[J]. Acta Microbiologica Sinica, 2019, 59(12): 2401-2409.(in Chinese with English abstract)
[28] 吴阔, 罗华元, 林昆, 等. 基于rDNA-ITS法鉴定的霉变烟叶真菌种类[J]. 贵州农业科学, 2014, 42(6): 98-100.
WU K, LUO H Y, LIN K, et al.Fungi species in mildew tobacco leaves identified based on rDNA-ITS[J]. Guizhou Agricultural Sciences, 2014, 42(6): 98-100.(in Chinese with English abstract)
[29] 赵文姬. 云南仓储烟叶霉变及其生物防治研究[D]. 北京: 中国农业科学院, 2004.
ZHAO W J.Tobacco mold and its biological control in the storage in Yunnan Province[D]. Beijing: Chinese Academy of Agricultural Sciences, 2004.(in Chinese with English abstract)
[30] 朱桂宁. 广西烟仓主要霉变生物的鉴定、生物学特性及防治研究[D]. 南宁: 广西大学, 2005.
ZHU G N.Studies on the fungi causing mildew of stored tobacco in Guangxi[D]. Nanning: Guangxi University, 2005.(in Chinese with English abstract)
[31] 曾婷英, 顾钢, 张绍升. 烘烤期烟叶霉烂病的病原鉴定[J]. 中国烟草学报, 2014, 20(4): 65-68.
ZENG T Y, GU G, ZHANG S S.Identification of tobacco mildew pathogen which causes leaf rot during flue-curing[J]. Acta Tabacaria Sinica, 2014, 20(4): 65-68.(in Chinese with English abstract)
[32] WANG H C, HUANG Y F, TANG X G, et al.Leaf and stem rot of tobacco (Nicotiana tabacum) caused by Rhizopus oryzae in closed curing barns in Guizhou Province of China[J]. Plant Disease, 2016, 100(2): 536.
[33] 祝明亮. 烟草调制期间微生物研究进展[J]. 微生物学通报, 2008, 35(8): 1278-1281.
ZHU M L.Advances in microbes during tobacco curing[J]. Microbiology, 2008, 35(8): 1278-1281.(in Chinese with English abstract)

Fungal composition and diversity analysis of healthy and rotten tobacco leaves after curing

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 9

Recommended Articles

Metrics

Comments

[1]	LUO Manli, LAN Qin, WANG Ge, WEI Hong, XIAO Jiujin, ZHANG Jian. Effect of fertilization on soil fauna community structure in farmland [J]. , 2019, 31(6): 946-954.
[2]	GOU Liqiong, YAO Heng, WANG Ge, HUAGN Rucheng, DUAN Junhua, XIAO Jiujin, ZHANG Jian. Effects of different straw returning methods on cropland soil fauna community [J]. , 2019, 31(3): 450-457.
[3]	GUI Guohong, YANG Hua, ZHU Jiangqun, ZHU Jianfeng, XIAO Yingping, XU E. Study on microbial community structure in chilled chicken during cold storage [J]. , 2019, 31(1): 47-55.
[4]	CHEN Yun, LIU Qi, DENG Junliang, YANG Yanyi, GAO Shuang, CHEN Chong, YAO Shuhua. Effects of composite antimicrobial peptide on rumen bacteria community structure of goat [J]. , 2017, 29(11): 1800-1808.
[5]	LI Yuan-yuan, SHI Kai, DELIGEER. Community structure and fauna of Lygus complex in Inner Mongolia [J]. , 2016, 28(9): 1558-1563.
[6]	ZHANG Aiju1，2，3， LIU Jindian1，2，3,，YANG Yuanjie1，2，3，GUO Aihuan1，2，3， GU Zhimin1，2，3，. Analysis of community characteristics of macrozoobenthos in enhancement and releasing zone in Tonglu section of Qiantang River [J]. , 2016, 28(8): 1323-.
[7]	LIU Dan, WU Feng-zhi. Effect of phenylalanine ammonialyase transgenic Arabidopsis thaliana on bacterial community in rhizosphere soil [J]. , 2016, 28(12): 2068-2075.
[8]	HAN Li-na, DING Zhe-li, ZENG Hui-cai, ZHENG Wei, HE Ying-dui, GE Yu. Effect of functional organic fertilizer on growth of Chinese cabbage [J]. , 2016, 28(10): 1718-1723.
[9]	YU You-ping;CAI Xin-zhong;ZHU Xiao-xiang;*. Comparative analysis of microbial community structures in soils from rice-upland crop rotation fields by PLFA profile technique [J]. , 2013, 25(5): 0-1061.