Difference Analysis of Fungal Community Among Bulk Soil,Rhizosphere and Rhizomes of Ligusticum chuanxiong Hort.

doi:10.13560/j.cnki.biotech.bull.1985.2020-0702

Abstract

Abstract:

It is aimed to obtain and compare the characteristics of fungal communities in bulk soil,rhizosphere and endophytic fungi of rhizome of Ligusticum chuanxiong Hort. High-throughput sequencing technology was used to analyze the structures and differences of fungal communities in bulk soil（S）,rhizosphere（R）and endophytic fungi（P）of the rhizome of L. chuanxiong Hort. FUNGuild was used to analyze the functions of fungal communities. There were high abundant fungal communities in the bulk soil,rhizosphere and endophytic fungi of the rhizome of L. chuanxiong Hort. The evenness and diversity of the fungal community showed as S>R>P,while the richness showed as R>S>P. Specific dominated fungal flora varied in the three groups at different taxonomic levels. Meanwhile,the fungal communities were mainly affected by the sample type,with the largest differences between bulk soil and rhizome,followed by that between rhizosphere and plants,and the least among the soils. Differences among origin areas were observed in the three groups separately,and ranked as S>R>P. Besides,the effect of environmental factors on the communities of the bulk soil and rhizosphere was S>R. The fungal community in the bulk soil was dominated by saprotroph,followed by pathotroph;while the rhizosphere soil was dominated by pathotroph,followed by saprotroph;and the endophytic fungi was dominated by pathotroph. The fungal structural characteristics and ecological functions presented continuous and gradual changes in the bulk soil to the rhizosphere soil and then to the rhizome of L. chuanxiong Hort.,which proved that the plants obtained special fungi from the soil during the cultivation process,continuous accumulation and reduction of these fungi occurred,and then special endophytic fungal community was formed. This way may be an important strategy for the construction of its microecological community structure and provide a scientific reference for understanding the formation of the authentic quality of Chinese medicine.

Key words: Ligusticum chuanxiong Hort., high-throughput sequencing, community diversity, rhizosphere, endophytic fungi

ZHANG Shu-hua, FANG Qian, JIA Hong-mei, HAN Gui-qi, YAN Zhu-yun, HE Dong-mei. Difference Analysis of Fungal Community Among Bulk Soil,Rhizosphere and Rhizomes of Ligusticum chuanxiong Hort.[J]. Biotechnology Bulletin, 2021, 37(4): 56-69.

Figures/Tables 13

References 33

[1]	张少鹏, 胥婷, 杨丽强, 等. 不同草原类型针茅根部内生真菌群落结构[J]. 应用生态学报, 2014(12):98-105.
	Zhang SP, Xu T, Yang LQ, et al. Endophytic fungal communities of Stipa sp. roots in different types of steppes in northern China[J]. Chinese Journal of Applied Ecology, 2014(12):98-105.
[2]	付静, 刘堰凤, 梅眉, 等. 韭菜根际真菌和内生真菌种群多样性分析[J]. 华北农学报, 2018,33(S1):250-256.
	Fu J, Liu YF, Mei M, et al. Comparing analysis of diversity of Chinese Leek rhizospheritic and endophytic fungal communities[J]. Acta Agriculturae Boreali-Sinica, 2018,33(S1):250-256.
[3]	Cordovez V, Diniandreote F, Carrion VJ, et al. Ecology and evolution of plant microbiomes[J]. Annual Review of Microbiology, 2019,73(1):69-88. doi: 10.1146/annurev-micro-090817-062524 URL
[4]	李毳, 刘怡. 晋东南3种道地药材植物根际真菌群落特性[J]. 生态环境学报, 2019,28(7):1388-1393.
	Li C, Liu Y, Characteristics of rhizosphere fungal community of three genuine medicinal plants in southeast Shanxi province[J]. Ecology and Environmental Sciences, 2019,28(7):1388-1393.
[5]	杨娟. 杜仲相关真菌群落组成及其对活性成分的影响[D]. 贵阳:贵州大学, 2019.
	Yang J. Community composition of the related fungi from Eucommia ulmoides Oliv. and its active constituents[D]. Guiyang:Guizhou University, 2019.
[6]	吴林坤, 黄伟民, 王娟英, 等. 不同连作年限野生地黄根际土壤微生物群落多样性分[J]. 作物学报, 2015,41(2):308-317.
	Wu LK, Huang WM, Wang JY, et al. Diversity analysis of rhizosphere microflora of wild R. glutinosa grown in monocropping for different years[J]. Acta Agronomica Sinica, 2015,41(2):308-317.
[7]	李吉. 怀牛膝连作对根际土壤微生物群落结构和功能多样性的影响[D]. 福州:福建农林大学, 2013.
	Li J. The changes in rhizospheric microbial community structure and functional diversity of Achyranthes Bidentata mediated by consecutive monoculture[D]. Fuzhou:Fujian Agriculture and Forestry University, 2013.
[8]	张培, 赵林华, 张海宇, 等. 川芎临床应用及其用量[J]. 吉林中医药, 2019,39(3):309-312.
	Zhang P, Zhao LH, Zhang HY, et al. Exploration about the clinical application and dosage of the rhizome of Chuanxiong[J]. Jilin Journal of Traditional Chinese Medicine, 2019,39(3):309-312.
[9]	刘圆, 贾敏如. 川芎品种、产地的历史考证[J]. 中药材, 2001,24(5):364-367.
	Liu Y, Jia MR. Historical investigation on the medicinal species and habitats of Ligusticum chuanxiong Hort.[J]. Traditional Chinese Medicine, 2001,24(5):364-367.
[10]	银玲, 彭月, 陈鸿平, 等. 新老产地川芎中酚酸类成分的含量比较及川芎等级划分内涵研究[J]. 中国实验方剂学杂志, 2013,19(11):91-94.
	Yin L, Peng Y, Chen HP, et al. Comparison of the content of Phenolic acids in Ligusticum chuanxiong from new and traditional producing areas and the connotation of division grades[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2013,19(11):91-94.
[11]	陈林, 刘友平, 陈鸿平, 等. 电子鼻在川芎不同产地不同等级评价中的应用[J]. 中药与临床, 2013(4):7-10.
	Chen L, Liu YP, Chen HP, et al. Electronic Nose used in evaluating Ligusticum chuanxiong Hort. produced in different regions and of different levels[J]. Pharmacy and Clinics of Chinese Materia Medica, 2013(4):7-10.
[12]	王光志, 吴清华, 李萍. 川芎根际土壤微生物多样性初步研究[C]. 中医药现代化国际科技大会, 2013.
	Wang GZ, Wu QH, Li P. Microbial communities diversity in rhizosphere soils of Ligusticum chuanxiong Hort. by PCR-DGGE[C]. International Conference on Modernization of Traditional Chinese Medicine, 2013.
[13]	汪杨丽, 严铸云, 郭晓恒, 等. 川芎内生真菌的分离与鉴定[J]. 中国中药杂志, 2008,33(9):999-1001.
	Wang LY, Yan ZY, Guo YH, et al. Isolation and identification of endophytic fungi from Ligusticum[J]. China Journal of Chinese Materia Medica, 2008,33(9):999-1001.
[14]	王海, 严铸云, 何冬梅, 等. 多产区川芎内生真菌菌群组成的PCR-DGGE分析[J]. 中国中药杂志, 2013,38(12):1893-1897.
	Wang H, Yan ZY, He DM, et al. Analysis of endophytic fungi community of Ligusticum chuanxiong using PCR-DGGE[J]. China Journal of Chinese Materia Medica, 2013,38(12):1893-1897.
[15]	严铸云, 汪杨丽, 郭晓恒, 等. 川芎内生真菌种群与川芎油的相关性分析[C]. 植物内生菌开发与应用学术研讨会, 2009.
	Yan ZY, Wang YL, Guo XH, et al. Correlation analysis of endophytic fungi community and volatile oil of Ligusticum chuanxiong[C]. Symposium on Development and Application of Plant Endophytes, 2009.
[16]	李佳穗, 严铸云, 兰英, 等. 四川主产区川芎根腐病病原菌鉴定[J]. 中药材, 2015,38(3):443-446.
	Li JS, Yan ZY, Lan Y, et al. Identification of pathogens causing root rot disease on Ligusticum chuanxiong in Sichuan[J]. Journal of Chinese Medicinal Materials, 2015,38(3):443-446.
[17]	何冬梅, 王海, 严铸云. 从川芎苓子中筛选根腐病拮抗性内生真菌的研究[EB]. 北京:中国科技论文在线 [2017-05-17].
	He DM, Wang H, Yan ZY. Screening of antagonistic endophytic fungi to root rot of Ligusticum chuanxiong Hort. [EB]. Beijing:Sciencepaper Online [2017-05-17].
[18]	胡容平, 龚国淑, 叶慧丽, 等. 川芎根腐病防治初步研究[J]. 西南农业学报, 2013,26(5):1873-1877.
	Hu RP, Gong GS, Ye HL, et al. Preliminary study on control effect to root rot disease of Ligusticum chuanxiong[J]. Southwest China Journal of Agricultural Sciences, 2013,26(5):1873-1877.
[19]	何冬梅. 川芎山地育苓的微生态机制研究[D]. 成都:成都中医药大学, 2016.
	He DM. Mechanism study on microbial ecology of mountain-breeding nodes of Ligusticum chuanxiong Hort.[D]. Chengdu:Chengdu University of Traditional Chinese Medicine, 2016.
[20]	鲍士旦. 土壤农化分析[M]. 北京: 中国农业出版社, 2000.
	Bao SD. Soil agrochemical analysis[M]. Beijing: China Agricultural Press, 2000.
[21]	杨剑虹, 王成林, 代亨林. 土壤农化分析与环境监测[M]. 北京: 中国大地出版社, 2008.
	Yang JH, Wang CL, Dai HL. Soil agrochemical analysis and environmental monitoring[M]. Beijing: China Land Press, 2008.
[22]	中华人民共和国农业部. NY/T1121. 7-2014 中华人民共和国农业行业标准[S]. 北京: 中国农业出版社, 2005.
	Ministry of agriculture of the people’s Republic of China. NY/T1121. 7-2014 Agricultural industry standard of the People’s Republic of China[S]. Beijing: China Agricultural Press, 2005.
[23]	中华人民共和国农业部. NY/T 889-2004 中华人民共和国农业行业标准[S]. 北京: 中国农业出版社, 2005.
	Ministry of agriculture of the People’s Republic of China. NY/T 889-2004 Agricultural industry standard of the People’s Republic of China[S]. Beijing: China Agricultural Press, 2005.
[24]	国家林业局. LY/T 1228-2015. 中华人民共和国林业行业标准[S]. 北京: 国家标准出版社, 2015.
	The state forestry administration of the People’s Republic of China LY/T 1228-2015 LY/T 1228-2015. Forestry industry standards of the People’s Republic of China[S]. Beijing: National Standards Press, 2015.
[25]	李莹, 秦明一, 张超, 等. 黄瓜内生真菌对根结线虫生防活性测定[J]. 湖北农业科学, 2016,55(3):634-637.
	Li Y, Qin MY, Zhang C, et al. Biocontrol against Meloidogyne incognita by an endophytic fungus Plectosphaerella sp. from cucumber root galls[J]. Hubei Agricultural Sciences, 2016,55(3):634-637.
[26]	Atkins SD, Clark IM, Sosnowska D, et al. Detection and quantification of Plectosphaerella cucumerina, a potential biological control agent of potato cyst nematodes, by using conventional PCR, Real-Time PCR, selective media, and baiting[J]. Applied and Environmental Microbiology, 2003,69(8):4788-4793. doi: 10.1128/AEM.69.8.4788-4793.2003 URL
[27]	Glynou K, Nam B, Thines M, et al. Facultative root-colonizing fungi dominate endophytic assemblages in roots of nonmycorrhizal Microthlaspi species[J]. New Phytologist, 2018,217(3):1190-1202. doi: 10.1111/nph.14873 URL
[28]	Nguyen NH, Song Z, Bates ST, et al. FUNGuild:An open annotation tool for parsing fungal community datasets by ecological guild[J]. Fungal Ecology, 2016,20:241-248. doi: 10.1016/j.funeco.2015.06.006 URL
[29]	Noah, Fierer. Embracing the unknown:disentangling the complexities of the soil microbiome[J]. Nature reviews Microbiology, 2017,15(10):579-590. doi: 10.1038/nrmicro.2017.87 URL
[30]	Zhu L, Wei Z, Yang T, et al. Core microorganisms promote the transformation of DOM fractions with different molecular weights to improve the stability during composting[J]. Bioresource Technology, 2020,299:122575. doi: 10.1016/j.biortech.2019.122575 URL
[31]	汪杨丽, 严铸云. 川芎内生真菌种群稳定性研究[J]. 时珍国医国药, 2009,20(12):2948-2949.
	Wang YL, Yan ZY. Population stability of endophytic fungi in Ligusticum chuanxiong Hort[J]. Lishizhen Medicine and Materia Medica Research, 2009,20(12):2948-2949.
[32]	Tedersoo L, Bahram M, Plme S, et al. Fungal biogeography. Global diversity and geography of soil fungi[J]. Science, 2014,346(6213):1256688. doi: 10.1126/science.1256688 URL
[33]	Edwards J, Johnson C, Santos-Medellín C, et al. Structure, variation, and assembly of the root-associated microbiomes of rice[J]. Proceedings of the National Academy of Sciences of the United States of America, 2015,112(8):911-920. doi: 10.1073/pnas.1414592112 pmid: 25605935

产区 Origin area	样点名称 Sample name	地址Address	土壤类型Soil type	栽培方式Planting model	样品编号Sample No.			纬度Latitude（N°）	经度Longitude （E°）	海拔Altitude /m
产区 Origin area	样点名称 Sample name	地址Address	土壤类型Soil type	栽培方式Planting model	非根际土Bulk soil/S	根际土Rhizosphere soil/R	根茎Rhizome/P	纬度Latitude（N°）	经度Longitude （E°）	海拔Altitude /m
石羊（SY）	SY1	都江堰市石羊镇	灰壤土	农户散种	SSY1	RSY1	PSY1	30.84	103.67	619
石羊（SY）	SY2	都江堰市石羊镇	灰壤土	农户散种	SSY2	RSY2	PSY2	30.83	103.68	603
石羊（SY）	SY3	都江堰市石羊镇	灰壤土	农户散种	SSY3	RSY3	PSY3	30.84	103.67	577
石羊（SY）	SY4	都江堰市石羊镇	灰壤土	农户散种	SSY4	RSY4	PSY4	30.82	103.67	591
石羊（SY）	SY5	都江堰市石羊镇	灰壤土	农户散种	SSY5	RSY5	PSY5	30.84	103.65	574
石羊（SY）	SY6	都江堰市石羊镇	灰壤土	承包种植	SSY6	RSY6	PSY6	30.83	103.66	597
眉山（MS）	XW	眉山市修文镇	沙壤土	承包种植	SXW	RXW	PXW	30.01	103.70	398
眉山（MS）	YS	眉山市永寿镇	红壤土	农户散种	SYS	RYS	PYS	29.99	103.85	364
眉山（MS）	XJ	眉山市谢家镇	红黏土	农户散种	SXJ	RXJ	PXJ	30.22	103.79	394
彭州（PZ）	PZ	彭州市敖平镇	灰壤土	承包种植	SPZ	RPZ	PAP	30.10	103.99	560
崇州（CZ）	CZ	崇州市梓潼镇	灰壤土	农户散种	SCZ	RCZ	PCZ	30.75	103.67	563
邛崃（QL）	QL	邛崃市回龙镇	棕黏土	承包种植	SQL	RQL	PQL	30.30	103.68	435

产区 Origin area	样点名称 Sample name	地址Address	土壤类型Soil type	栽培方式Planting model	样品编号Sample No.			纬度Latitude（N°）	经度Longitude （E°）	海拔Altitude /m
产区 Origin area	样点名称 Sample name	地址Address	土壤类型Soil type	栽培方式Planting model	非根际土Bulk soil/S	根际土Rhizosphere soil/R	根茎Rhizome/P	纬度Latitude（N°）	经度Longitude （E°）	海拔Altitude /m
石羊（SY）	SY1	都江堰市石羊镇	灰壤土	农户散种	SSY1	RSY1	PSY1	30.84	103.67	619
石羊（SY）	SY2	都江堰市石羊镇	灰壤土	农户散种	SSY2	RSY2	PSY2	30.83	103.68	603
石羊（SY）	SY3	都江堰市石羊镇	灰壤土	农户散种	SSY3	RSY3	PSY3	30.84	103.67	577
石羊（SY）	SY4	都江堰市石羊镇	灰壤土	农户散种	SSY4	RSY4	PSY4	30.82	103.67	591
石羊（SY）	SY5	都江堰市石羊镇	灰壤土	农户散种	SSY5	RSY5	PSY5	30.84	103.65	574
石羊（SY）	SY6	都江堰市石羊镇	灰壤土	承包种植	SSY6	RSY6	PSY6	30.83	103.66	597
眉山（MS）	XW	眉山市修文镇	沙壤土	承包种植	SXW	RXW	PXW	30.01	103.70	398
眉山（MS）	YS	眉山市永寿镇	红壤土	农户散种	SYS	RYS	PYS	29.99	103.85	364
眉山（MS）	XJ	眉山市谢家镇	红黏土	农户散种	SXJ	RXJ	PXJ	30.22	103.79	394
彭州（PZ）	PZ	彭州市敖平镇	灰壤土	承包种植	SPZ	RPZ	PAP	30.10	103.99	560
崇州（CZ）	CZ	崇州市梓潼镇	灰壤土	农户散种	SCZ	RCZ	PCZ	30.75	103.67	563
邛崃（QL）	QL	邛崃市回龙镇	棕黏土	承包种植	SQL	RQL	PQL	30.30	103.68	435

样本类型Sample type	香农指数 Shannon index	辛普森指数Simpson index	Ace指数 Ace index	Chao指数 Chao index	Coverage指数Coverage index	Shannon even指数Shannon even index
非根际土Bulk soil（S）	4.6244a	0.0376c	1090.68b	1089.08b	0.9942b	0.6816a
根际土Rhizosphere soil（R）	3.9585b	0.0661b	1292.43a	1190.61a	0.9918c	0.5879b
川芎根茎Rhizome（P）	1.8919c	0.3401a	400.24c	355.53c	0.9976a	0.3431c

样本类型Sample type	香农指数 Shannon index	辛普森指数Simpson index	Ace指数 Ace index	Chao指数 Chao index	Coverage指数Coverage index	Shannon even指数Shannon even index
非根际土Bulk soil（S）	4.6244a	0.0376c	1090.68b	1089.08b	0.9942b	0.6816a
根际土Rhizosphere soil（R）	3.9585b	0.0661b	1292.43a	1190.61a	0.9918c	0.5879b
川芎根茎Rhizome（P）	1.8919c	0.3401a	400.24c	355.53c	0.9976a	0.3431c

门Phylum	非根际土Bulk soil（S）	根际土Rhizosphere soil（R）	川芎根茎Rhizome （P）
子囊菌门Ascomycota***	59.60%	71.43%	90.05%
担子菌门Basidiomycota***	14.52%	16.20%	7.843%
被孢菌门Mortierellomycota***	14.8%	8.397%	0.9983%
罗兹菌门Rozellomycota***	1.787%	0.7173%	0.0467%