Sequencing Analysis of Fungal Diversity in Rhizosphere Soil of Cynanchum bungei Decne

doi:10.13304/j.nykjdb.2021.0963

Abstract

Abstract:

To understand the structure and composition of rhizosphere soil microbial community of Cynanchum bungei Decne， and make full use of rhizosphere microbial resources， the fungal community richness and diversity in rhizosphere soil （at 10 and 20 cm deep） of Cynanchum bungei Decne form Lianhuashan of Laiwu City （LW）， Lixia （LX） and Changqing （CQ） of Jinan City of Shandong Province were analyzed by Illumina MiSeq high throughput sequencing technology. The results showed that 1 589 OTUs were obtained from 6 samples， belonging to 11 phyla， 35 classes， 84 orders， 142 families and 237 genera. Ascomycota and Mortierellomycota were the common dominant bacteria of 6 samples at phyla level. Sample CQ20 had the highest community abundance and diversity， and sample LX20 had the most uniform distribution of fungal community. With the deepening of underground rhizome depth， the abundance and diversity of fungal microbial community increased， which provided theoretical basis for optimizing the rhizosphere growth-promoting bacteria and improving the planting technique of simulating wild.

Key words: Cynanchum bungei Decne, rhizosphere soil, fungi, Illumina next-generation sequencing

摘要：

为了解泰山白首乌根际土壤微生物群落结构和组成，充分利用根际微生物资源，采用Illumina MiSeq高通量测序技术，分析测定来自山东省莱芜市莲花山（LW）和济南市历下区（LX）、长清区（CQ）3个产地的泰山白首乌根茎10和20 cm深度根际土壤真菌群落的丰富度和多样性。结果表明，6个样品共得到真菌1 589个OTUs，分属于11门35纲84目142科237属。子囊菌门（Ascomycota）和被孢霉门（Mortierellomycota）是6个样品共同的优势菌门。样品CQ20的群落丰度和多样性最高，样品LX20的真菌群落分布最均匀；随着地下根茎深度加深，真菌微生物群落的丰度和多样性增大。不同产地和根茎深度样品间土壤真菌群落存在显著差异，上述结果为优选根际促生菌、提高仿野生种植技术提供了理论依据。

关键词: 泰山白首乌, 根际土壤, 真菌, 高通量测序

CLC Number:

Jie LI, Ying LIN, Meiyu XU, Fei WANG, Lingchuan XU. Sequencing Analysis of Fungal Diversity in Rhizosphere Soil of Cynanchum bungei Decne[J]. Journal of Agricultural Science and Technology, 2022, 24(6): 70-81.

李洁, 林莹, 徐美玉, 王飞, 徐凌川. 泰山白首乌根际土壤真菌多样性分析[J]. 中国农业科技导报, 2022, 24(6): 70-81.

Figures/Tables 14

Table 1 Sequence results of rhizosphere soil of Cynanchum bungei Decne

样品 Sample	序列数 Sequence number	碱基数 Base number	平均长度 Mean length/bp	最小长度 Min. length/bp	最大长度 Max. length/bp	操作分类单元数OTUs
LW10	68 579	18 104 375	263.99	47	380	244
LW20	68 939	17 199 374	249.49	43	490	175
LX10	58 106	16 270 921	280.02	44	391	216
LX20	64 833	20 109 613	310.18	42	491	337
CQ10	49 236	14 888 556	302.39	42	490	259
CQ20	68 679	19 492 433	283.82	42	491	358

Fig. 1 Veen diagram of rhizosphere soil samples of Cynanchum bungei DecneA： 6 sample groups； B： Sample group underground 10 cm； C： Sample group underground 20 cm

Fig. 2 Dilution curve of rhizosphere soil sample of Cynanchum bungei Decne

Table 2 Statistical table of rhizosphere soil fungal diversity index of Cynanchum bungei Decne

样品Sample	Chao 指数 Chao index	Ace 指数 Ace index	Shannon指数Shannon index	Simpson指数Simpson index	Shannon均匀指数Shannon even index	覆盖度 Coverage
LW10	254.01	256.96	2.83	0.13	0.51	1.0
LW20	190.95	192.75	1.87	0.31	0.36	1.0
LX10	227.11	227.59	2.39	0.23	0.44	1.0
LX20	340.75	342.87	3.71	0.07	0.64	1.0
CQ10	264.53	264.62	3.50	0.10	0.63	1.0
CQ20	361.75	363.10	3.69	0.07	0.63	1.0

Fig. 3 Rank abundance curve of rhizosphere soil samples of Cynanchum bungei Decne

Fig. 4 Relationship of different rhizosphere soil samples

Fig. 5 Distance heat map of rhizosphere soil sample of Cynanchum bungei Decne

Fig. 6 PCA of different rhizosphere soil samples

Table 3 Number of Cynanchum bungei Decne samples at different classification levels

分类Taxonomy	LW10	LW20	LX10	LX20	CQ10	CQ20
门Plylum	9	9	8	10	8	9
纲Class	18	17	21	29	23	24
目Order	44	40	53	60	49	58
科Family	73	58	85	106	86	106
属Genus	104	75	114	142	130	169
种Species	167	116	171	229	196	268

Fig. 7 Relative abundance of fungi in different rhizosphere soil samples at phylum level

Fig. 8 Relative abundance of fungi in different rhizosphere soil samples at class level

Fig. 9 Relative abundance of fungi in different rhizosphere soil samples at order level

Fig. 10 Relative abundance of fungi in different rhizosphere soil samples at family level

Fig. 11 Relative abundance of fungi in different rhizosphere soil samples at genus level

References 34

1	印鑫,丁永芳,邵久针,等.白首乌的研究进展[J].中草药,2019,50(4):992-1000.
	YIN X, DING Y F, SHAO J Z, et al.. Research progress on Cynanchi Bungei Radix [J]. Chin. Trad. Herbal Drugs, 2019, 50(4):992-1000.
2	刘佳,牛景梅,赵进红,等.泰山白首乌研究进展[J].广东化工,2019,46(6):119-120.
	LIU J, NIU J M, ZHAO J H, et al.. Research progress of Cynanchum bungei Decne [J]. Guangdong Chem. Ind., 2019, 46(6):119-120.
3	陈东伟,胡明哲,李克明,等.泰山白首乌的药理作用和临床应用[J].临床医药文献电子杂志,2017,4(54):10666-10667.
4	庄子锐,王明亮,彭蕴茹,等.白首乌C21甾苷通过TLR4通路防治大鼠肝肾纤维化的作用研究[J].中国中药杂志,2021,46(11):2857-2864.
	ZHUANG Z R, WANG M L, PENG Y R, et al.. Effect of C21 steroidal glycoside of Cynanchi Bungei Radix on liver and kidney fibrosis in rats through TLR4 pathway [J]. Chin. J. Trad. Chin. Med., 2021, 46(11):2857-2864.
5	牛景梅,吕宝兴,赵进红,等.泰山白首乌中白首乌二苯酮在大鼠体内药代动力学研究[J].泰山医学院学报,2020,41(5):349-352.
	NIU J M, LV B X, ZHAO J H, et al.. Pharmacokinetics of diphenyl ketone from Cynanchum bungei Decne in rats [J]. J. Taishan Med. Coll., 2020, 41(5):349-352.
6	张明,顾小兵,李春阳,等.白首乌水旱轮作种植技术研究[J].农业开发与装备,2020(11):197-198.
	ZHANG M, GU X B, LI C Y, et al.. Study on water and drought rotation planting technology of Cynanchum bungei Decne [J]. Agric. Develop. Equip., 2020(11):197-198.
7	毛爱华,周丽娟,毛广才.“苏乌1号”白首乌高产高效配套栽培技术[J].南方农业,2019,13(35):31-32, 34.
	MAO A H, ZHOU L J, MAO G C. High yield and high efficiency cultivation techniques of “Suwu No. 1” Cynanchum bungei Decne [J]. Southern Agric., 2019, 13(35):31-32, 34.
8	吴承东,张明.几种土壤封闭除草剂对白首乌田的除草效果试验[J].农业开发与装备,2017(11):94.
	WU C D, ZHANG M. Effects of several soil blocking herbicides on Baishouwu field [J]. Agric. Develop. Equip., 2017(11):94.
9	王朋强,吴义兰,李永富,等.植物根系分泌物及微域环境的研究与展望[J].乡村科技,2021,12(27):96-98.
10	唐杰,陈知青,郭安南,等.不同作物根际土壤微生物的群落结构特征分析[J].核农学报,2021,35(12):2830-2840.
	TANG J, CHEN Z Q, GUO A N, et al.. Community structure characteristics of soil microorganisms in rhizosphere of different crops [J]. J. Nuclear Agric., 2021, 35(12):2830-2840.
11	杨露,辛建攀,田如男.根际微生物对植物重金属胁迫的缓解作用及其机理研究进展[J].生物技术通报,2022,38(3):213-225.
	YANG L, XIN J P, TIAN R N. Research progress on mitigation effect and mechanism of rhizosphere microorganisms on plant heavy metal stress [J]. Biotechnol. Bull., 2022,38(3):213-225.
12	连文慧,董雷,李文均.土壤环境下的根际微生物和植物互作关系研究进展[J].微生物学杂志,2021,41(4):74-83.
	LIAN W H, DONG L, LI W J. Research progress on interaction between rhizosphere microorganisms and plants in soil environment [J]. J. Microbiol., 2021, 41(4):74-83.
13	任春光,苏文文,潘丽珊,等.基于高通量测序研究猕猴桃苗不同生育期根际真菌群落结构及多样性[J].土壤,2021,53(3):545-554.
	REN C G, SU W W, PAN L S, et al.. Study on rhizosphere fungal community structure and diversity of kiwifruit seedlings at different growth stages based on high throughput sequencing [J]. Soils, 2021, 53(3):545-554.
14	付丽娜,汪娅婷,王星,等.三七连作根际微生物多样性研究[J].云南农业大学学报(自然科学),2018,33(2):198-207.
	FU L N, WANG Y T, WANG X, et al.. Study on microbial diversity in rhizosphere of Panax notoginseng continuous cropping [J]. J. Yunnan Agric. Univ. (Nat. Sci.), 2018, 33(2):198-207.
15	张艳琪,周甜甜,彭川岳,等.泰山白首乌根际土壤真菌分离纯化及抗菌活性研究[J].中国抗生素杂志,2021,46(6):552-556.
	ZHANG Y Q, ZHOU T T, PENG C Y, et al.. Isolation, purification and antibacterial activity of rhizosphere soil fungi from Cynanchum bungei Decne [J]. Chin. J. Antibiotics, 2021, 46(6):552-556.
16	CHEN W H, ZHANG Z Z, BAN Y F, et al.. Cynanchum bungei Decne and its two related species for “Baishouwu”: a review on traditional uses, phytochemistry, and pharmacological activities [J/OL]. J. Ethnopharmacol., 2019, 243:112110 [2021-10-05]. .
17	顾晓洁,解卓学,吕嘉东,等.白首乌内生细菌分离鉴定及系统发育树分析[J].辽宁中医药大学学报,2018,20(6):52-56.
	GU X J, XIE Z X, LV J D, et al.. Isolation, identification and phylogenetic tree analysis of endophytic bacteria from Cynanchum bungei Decne [J]. J. Liaoning Univ. TCM, 2018, 20(6):52-56.
18	杨光柱,黄文静,郑丽萍,等.基于高通量测序的苹果根腐病病株和健株根际土壤细菌组成与多样性分析[J].西南农业学报,2021,34(9):1865-1869.
	YANG G Z, HUANG W J, ZHENG L P, et al.. Analysis of bacterial composition and diversity in rhizosphere soil of diseased and healthy apple root rot plants based on high throughput sequencing [J]. J. Southwest Agric., 2021, 34(9):1865-1869.
19	刘元,李守婷,申进文,等.基于GSS高通量测序技术的怀山药根、茎中内生真菌群落多样性分析[J].浙江农业科学,2018,59(1):64-68.
	LIU Y, LI S T, SHEN J W, et al.. Diversity analysis of endophytic fungi community in roots and stems of Huaishan herb based on GSS high throughput sequencing technology [J]. Zhejiang Agric. Sci., 2018, 59(1):64-68.
20	吴秋芳,侯立江,何玲敏,等.北艾根际与非根际土壤微生物多样性的高通量测序分析[J].河南农业大学学报,2021,55(5):928-935.
	WU Q F, HOU L J, HE L M, et al.. High throughput sequencing analysis of microbial diversity in rhizosphere and non rhizosphere soil of Artemisia argyi [J]. J. Henan Agric.Univ., 2021, 55(5):928-935.
21	程立君,王磊,吴银梅,等.高通量测序技术分析光叶珙桐根际土壤微生物多样性[J].广东农业科学,2019,46(1):43-49.
	CHENG L J, WANG L, WU Y M, et al.. Analysis of rhizosphere soil microbial diversity of Davidia involucrata by high throughput sequencing [J]. Guangdong Agric. Sci., 2019, 46(1):43-49.
22	翁志强,许钰滢,吕正阳,等.巨菌草根际土壤微生物群落代谢功能多样性分析[J].广东农业科学,2021,48(4):62-68.
	WENG Z Q, XU Y Y, LYU Z Y, et al.. Analysis on metabolic function diversity of soil microbial community in rhizosphere of giant fungus grass [J]. Guangdong Agric. Sci., 2021, 48(4):62-68.
23	王海斌,陈晓婷,丁力,等.连作茶树根际土壤自毒潜力、酶活性及微生物群落功能多样性分析[J].热带作物学报,2018,39(5):852-857.
	WANG H B, CHEN X T, DING L, et al.. Analysis of autotoxic potential, enzyme activity and functional diversity of microbial community in rhizosphere soil of continuous cropping tea [J]. J. Tropical Crops, 2018, 39(5):825-857.
24	陈冉,刘志强,王丹丹.基于高通量测序比较不同产地药用银杏根际土壤微生物多样性[J].药物生物技术,2021,28(2):117-122.
	CHEN R, LIU Z Q, WANG D D. Comparison of microbial diversity in rhizosphere soil of medicinal Ginkgo biloba from different habitats based on high throughput sequencing [J]. Pharm Biotechnol., 2021, 28(2):117-122.
25	官鑫,向阳,祝友朋,等.核桃根际土壤微生物群落功能多样性分析[C]// 中国植物病理学会.中国植物病理学会2019年学术年会论文集.北京:中国农业科学技术出版社,2019:1.
26	郑雪芳,苏远科,刘波,等.不同海拔茶树根系土壤微生物群落多样性分析[J].中国生态农业学报,2010,18(4):866-871.
	ZHENG X F, SU Y K, LIU B, et al.. Diversity analysis of soil microbial communities in tea roots at different altitudes [J]. Chin. J. Ecol. Agric., 2010, 18(4):866-871.
27	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 communities of three genuine medicinal plants in Southeast Shanxi [J]. J. Ecol. Environ., 2019, 28(7):1388-1393.
28	SHEN C C, SHI Y, NI Y Y, et al.. Dramatic increases of soil microbial functional gene diversity at the treeline ecotone of Changbai Mountain [J/OL]. Front Microbiol., 2016,7:1184 [2021-07-15]. .
29	褚海燕,冯毛毛,柳旭,等.土壤微生物生物地理学:国内进展与国际前沿[J].土壤学报,2020,57(3):515-529.
	ZHE H Y, FENG M M, LIU X, et al.. Soil microbial biogeography: domestic progress and international frontier [J]. J. Soil Sci., 2020, 57(3):515-529.
30	ZHANG Y G, LIU X, CONG J, et al.. The microbially mediated soil organic carbon loss under degenerative succession in an alpine meadow [J]. Mol. Ecol., 2017, 26(14):3676-3686.
31	付亚娟,张江丽,侯晓强.大花杓兰根际与非根际土壤真菌多样性的高通量测序分析[J].西北农业学报,2019, 28(2):253-259.
	FU Y J, ZHANG J L, HOU X Q. High-throughput sequencing analysis of fungal diversity in rhizosphere and non-rhizosphere soils of Cyclobalanopsis grandiflora [J]. J. Northwest Agric., 2019, 28(2):253-259.
32	卢宝慧,高成林,赵玥,等.运用高通量测序技术分析人参不同栽培模式根际土壤微生物多样性[J].东北林业大学学报,2021,49(3):113-119.
	LU B H, GAO C L, ZHAO Y, et al.. Analysis of microbial diversity in rhizosphere soil under different cultivation patterns of ginseng by high-throughput sequencing [J]. J. Northeast For.Univ., 2021, 49(3):113-119.
33	祝蕾,严辉,刘培,等.药用植物根际微生物对其品质形成的影响及其作用机制的研究进展[J].中草药,2021,52(13):4064-4073.
	ZHU L, YAN H, LIU P, et al.. Research progress on the effect of rhizosphere microorganisms on the quality formation of medicinal plants and its mechanism of action [J]. Chin. Herbal Medic., 2021, 52(13):4064-4073.
34	孟祥才,沈莹,杜虹韦.道地药材概念及其使用规范的探讨[J].中草药,2019,50(24):6135-6141.
	MENG X C, SHEN Y, DU H W. Discussion on the concept of genuine medicinal materials and their use specifications [J]. Chin. Tradit. Herbal Drugs, 2019, 50(24):6135-6141.

[1]	Hongyan ZHAO, Junwei TAN, Jie ZHANG, Haonan CHEN, Chunxu WANG, Di ZHAO, Haipeng LI, Lixia ZHU, Yiqiang HAN. Community Structure of Stem-based Fungi Infected with Adzuki Bean and Mung Bean [J]. Journal of Agricultural Science and Technology, 2022, 24(5): 129-136.
[2]	ZHANG Qing1,2, ZHAO Xiaomei2, WANG Ping2, LIU Bing1, CHEN Ying2. Advance on Detection Methods of Nucleic Acid for Toxin-Producing Fungi in Food [J]. Journal of Agricultural Science and Technology, 2021, 23(3): 91-98.
[3]	YIN Quanyu1, LIU Jianhao1, LIU Guoshun1, YANG Xinling2, LI Xiaofu2, ZHANG Yulan1, LI Yang1,YE Hongchao3*. Effects of Biochar Application for Four Consecutive Years on Microbial Community Structure of Tobacco Cinnamon Soil [J]. Journal of Agricultural Science and Technology, 2021, 23(1): 176-185.
[4]	ZHAO Hu1,2, WANG Haibin2,3, CHEN Xiaoting2,3, WANG Yuhua2, ZHANG Huabin2, DING Li2, KONG Xianghai2, SHI Guiying1. Analysis of Nitrogen Composition and its Absorption and Utilization Efficiency in Rhizosphere Soil of Tea Tree [J]. Journal of Agricultural Science and Technology, 2020, 22(7): 148-153.
[5]	YANG Jingjing, ZHANG Qingqing*, Tuerxunnayi·Reyimu, Amanula·Yimingniyazi, Xueretijiang·Maitinuri. Effects of Nomadic Grazing and Settled Grazing on the Diversity of Fungi Community in Seriphidium transiliense Desert Grassland [J]. Journal of Agricultural Science and Technology, 2020, 22(7): 166-173.
[6]	WANG Junfeng1§, WEN Xiaolei1§, SUN Weiming1, LIU Yijian1, SHI Luting2, ZHANG Nana1, YANG Wenjie1, CHEN Weidong3, QI Huixia1*. Study on Cultural Conditions of Penicillium spinulosum Causing Chestnut Rot and Fungicides Screening [J]. Journal of Agricultural Science and Technology, 2020, 22(3): 79-84.
[7]	JIANG Caige1, SONG Shuang1, LI Qian2, WANG Guozhen1, ZHANG Yi1*, GUAN Xiaoyan3. Evaluation of Resistance Levels of Botrytis cinerea to Different Fungicides in Eastern Helan Mountain [J]. Journal of Agricultural Science and Technology, 2020, 22(10): 119-128.
[8]	GAO Lin1, WANG Xinwei1, SHEN Guoming1, TIAN Feng2, CHEN Qianfeng2, ZHANG Mingfa2, ZHANG Chengsheng1*. Differences of Bacteria and Fungi Community Structure in Tobacco-planting Soil of Different Continuous Cropping Years [J]. Journal of Agricultural Science and Technology, 2019, 21(8): 147-152.
[9]	SHI Fangfang, LI Xiangquan*. Diversity Analysis of Fungus Community in Rhizosphere Soil of Grape [J]. Journal of Agricultural Science and Technology, 2019, 21(7): 47-58.
[10]	GU Meiying1§, YANG Rong1§, XU Wanli2*, TANG Guangmu2, ZHANG Zhidong1, ZHANG Yunshu2, FENG Lei2, WANG Ning1, ZHAN Faqiang1. Effects of Cotton Stalk Biochar Combined with Bio-organic Fertilizer on Rhizosphere Soil Micro-ecology and Cotton Growth of Continuous Cropping Cotton [J]. Journal of Agricultural Science and Technology, 2019, 21(10): 47-57.
[11]	HUANG Xiaoyun1, SHEN Huawei2, HAN Haidong1, ZHONG Zhenmei1, LUO Tao1, HUANG Xiusheng1*. Research Progress and Countermeasures on Recycling Utilization Model of Edible Fungi Industry By-products [J]. Journal of Agricultural Science and Technology, 2019, 21(10): 125-132.
[12]	ZHANG Yijie, SHAO Huifang*, ZHANG Ke, JIA Hongfang, HUANG Wuxing, HAN Dan. Influences of Fertilization on Soil Environment and Microorganism in Continuous Cropping Based on High-Throughput Sequencing [J]. Journal of Agricultural Science and Technology, 2018, 20(5): 16-25.
[13]	GONG Zhixiang1, MA Xiaohan1, REN Zhiguang1, ZHU Jinfeng2,HUANG Yuanjiong3, WANG Mengmeng1, CHEN Zheng1, XU Zicheng1*. Analysis of Bacterial Communities in Rhizosphere Soil of Continuous Cropping Flue-cured Tobacco Using 16S rDNA-PCR-DGGE [J]. Journal of Agricultural Science and Technology, 2018, 20(2): 39-47.
[14]	XU Fan1§, CAO Rui1§, CHEN Zhixing2, WANG Haining3, LI Fan1, WANG Yang1, XIE Yong1. Research on Nematicidal Effect and Integrative Utility of Marigold Stalk [J]. Journal of Agricultural Science and Technology, 2017, 19(7): 124-130.
[15]	GAO Yuan, WANG Fuling, JIA Qi, LIU Wei, ZHOU Yanyan, LI Jiali. Isolation and Identification of Endophytic Fungi from Taraxacum mongolicum Hand-Mazz and Antibacterial Activity Study [J]. Journal of Agricultural Science and Technology, 2017, 19(6): 55-60.