Composition Features of Microbial Community in the Rhizospheric Soil of Bletilla striata with Root Rot

doi:10.13560/j.cnki.biotech.bull.1985.2021-0757

Abstract

Abstract:

Analyzing the composition features of microbial community in the rhizospheric soil of Bletilla striata with root rot would provide the scientific basis for understanding the microbial mechanisms leading to the root rot of B. striata and taking corresponding prevention and control measures. The Illumina HiSeq high-throughput sequencing was used to compare and analyze the microbial community structure in the rhizospheric soils of root-rot and healthy B. striata. Also,the soil physicochemical properties and enzyme activities were quantitatively analyzed. The dominant fungi were Ascomycota and Mortierellomycota,and the dominant bacteria were Proteobacteria and Bacteroidetes in the rhizospheric soil of B. striata. Compared with the healthy individuals,the relative abundance of Ascomycota significantly increased,while the relative abundance of Mortierellomycota decreased. The relative abundance of pathogenic Fusarium and Erysiphe significantly increased in the rhizospheric soil of root-rot B. striata（P<0.05）. Multivariate statistical analyses showed the significant differentiation of bacterial and fungal community in the rhizospheric soils between healthy and root-rot B. striata. In addition,the activities of urease,protease and sucrase,and available potassium content were higher in the rhizospheric soil of healthy individuals;while soil pH and the contents of organic matter,NH₄⁺-N,and NO₃^--N were lower. The root-rot disease of B. striata could be related to the increased relative abundance of pathogenic microorganisms,the changes of microbial community structure,the decreased soil enzyme activity,and the imbalance of plant nutrients.

Key words: Bletilla striata, root-rot, soil microbial community, high-throughput sequencing

ZHAO Lin-yan, GUAN Hui-lin, XIANG Ping, LI Ze-cheng, BAI Yu-long, SONG Hong-chuan, SUN Shi-zhong, XU Wu-mei. Composition Features of Microbial Community in the Rhizospheric Soil of Bletilla striata with Root Rot[J]. Biotechnology Bulletin, 2022, 38(2): 67-74.

Figures/Tables 6

References 48

[1]	孙乐乐, 杨永红, 刘军凯, 等. 白及的本草考证[J]. 中药材, 2010, 33(12):1965-1968.
	Sun LL, Yang YH, Liu JK, et al. Research on Bletilla striata[J]. J Chin Med Mater, 2010, 33(12):1965-1968.
[2]	张曼, 韩亭亭, 胡春芳, 等. 白及产业现状及可持续发展策略[J]. 中草药, 2019, 50(20):5103-5108.
	Zhang M, Han TT, Hu CF, et al. Industrialization condition and sustainable development strategies of Bletillae Rhizoma[J]. Chin Tradit Herb Drugs, 2019, 50(20):5103-5108.
[3]	柯尚艳, 杨林毅, 等. 白及植株上一种真菌病害的分离与鉴定[J]. 云南农业大学学报:自然科学, 2018, 33(3):405-409.
	Ke SY, Yang LY, et al. Isolation and identification of a fungal disease from Bletilla striata[J]. J Yunnan Agric Univ:Nat Sci, 2018, 33(3):405-409.
[4]	龙小琴, 戴应和. 白及栽培技术要点及主要病虫害防治研究进展[J]. 农业与技术, 2020, 40(14):81-84.
	Long XQ, Dai YH. Main points of cultivation techniques and control of diseases of Bletilla striata[J]. Agric Technol, 2020, 40(14):81-84.
[5]	廖长宏, 陈军文, 吕婉婉, 等. 根和根茎类药用植物根腐病研究进展[J]. 中药材, 2017, 40(2):492-497.
	Liao CH, Chen JW, Lv WW, et al. Research on root rot disease of rhizome medicinal plant[J]. J Chin Med Mater, 2017, 40(2):492-497.
[6]	缪作清, 李世东, 刘杏忠, 等. 三七根腐病病原研究[J]. 中国农业科学, 2006, 39(7):1371-1378.
	Miao ZQ, Li SD, Liu XZ, et al. The causal microorganisms of Panax notoginseng root rot disease[J]. Sci Agric Sin, 2006, 39(7):1371-1378.
[7]	毕武, 陈娟, 焦晓林, 等. 北京地区西洋参根腐病病原鉴定及其致病性[J]. 植物保护, 2011, 37(5):135-138.
	Bi W, Chen J, Jiao XL, et al. Identification of the pathogens causing the root rot and their pathogenicity on American ginseng in Beijing[J]. Plant Prot, 2011, 37(5):135-138.
[8]	曾令祥, 杨琳, 陈娅娅, 等. 贵州中药材白及病虫害种类的调查与综合防治[J]. 贵州农业科学, 2012, 40(7):106-108.
	Zeng LX, Yang L, Chen YY, et al. Investigation and integrated management on diseases and pests for Bletilla striata in Guizhou[J]. Guizhou Agric Sci, 2012, 40(7):106-108.
[9]	曾茜, 陈旭, 刘璞玉, 等. 贵州小白及内生真菌对块茎腐烂病拮抗作用研究[J]. 南方农业, 2018, 12(1):5-7.
	Zeng QX, Chen X, Liu PY, et al. Antagonistic effects of endophytic fungi of Guizhou Bletilla striata on tuber rot[J]. South China Agric, 2018, 12(1):5-7.
[10]	Xiong W, Song Y, Yang K, et al. Rhizosphere protists are key determinants of plant health[J]. Microbiome, 2020, 8(1):27. doi: 10.1186/s40168-020-00799-9 pmid: 32127034
[11]	Philippot L, Raaijmakers JM, Lemanceau P, et al. Going back to the roots:the microbial ecology of the rhizosphere[J]. Nat Rev Microbiol, 2013, 11(11):789-799. doi: 10.1038/nrmicro3109 pmid: 24056930
[12]	She S, Niu J, Zhang C, et al. Significant relationship between soil bacterial community structure and incidence of bacterial wilt disease under continuous cropping system[J]. Arch Microbiol, 2017, 199(2):267-275. doi: 10.1007/s00203-016-1301-x URL
[13]	Garbeva P, van Veen JA, van Elsas JD. Microbial diversity in soil:selection microbial populations by plant and soil type and implications for disease suppressiveness[J]. Annu Rev Phytopathol, 2004, 42:243-270. pmid: 15283667
[14]	闫欢, 高芬, 王梦亮, 等. 黄芪根腐病病株和健株根围微生物菌群变化分析[J]. 植物保护, 2020, 46(4):48-54.
	Yan H, Gao F, Wang ML, et al. Changes of microbial community in root zone soil of Astragalus membranaceus suffering from root rot disease[J]. Plant Prot, 2020, 46(4):48-54.
[15]	谢玉清, 茆军, 王玮, 等. 大蒜根腐病根际土壤真菌群落结构及多样性分析[J]. 中国农学通报, 2020, 36(13):145-153.
	Xie YQ, Mao J, Wang W, et al. Structures and biodiversity of fungal communities in rhizosphere soil of root rot diseased garlic[J]. Chin Agric Sci Bull, 2020, 36(13):145-153.
[16]	Wu ZX, Hao ZP, Sun YQ, et al. Comparison on the structure and function of the rhizosphere microbial community between healthy and root-rot Panax notoginseng[J]. Appl Soil Ecol, 2016, 107:99-107. doi: 10.1016/j.apsoil.2016.05.017 URL
[17]	Jiang JL, Yu M, Hou RP, et al. Changes in the soil microbial community are associated with the occurrence of Panax quinquefolius L. root rot diseases[J]. Plant Soil, 2019, 438(1/2):143-156. doi: 10.1007/s11104-018-03928-4 URL
[18]	蒋景龙, 余妙, 等. 西洋参根腐病发生与根际土壤细菌群落结构变化关系研究[J]. 中草药, 2018, 49(18):4399-4407.
	Jiang JL, Yu M, et al. Relationship between occurrence of root-rot and changes of bacterial community structure in rhizosphere soil of Panax quinquefolius[J]. Chin Tradit Herb Drugs, 2018, 49(18):4399-4407.
[19]	鲁如坤. 土壤农业化学分析方法[M]. 北京: 中国农业科技出版社, 1999:146-195.
	Lu RK. Methods in analyzing agricultural soil chemistry[M]. Beijing: China Agricultural Science and Technology Press, 1999:146-195.
[20]	姚槐应, 黄昌勇. 土壤微生物生态学及其实验技术[M]. 北京: 科学出版社, 2006.
	Yao HY, Huang CY. Soil microbial ecology and experimental technology[M]. Beijing: Science Press, 2006.
[21]	赵兰坡, 姜岩. 土壤磷酸酶活性测定方法的探讨[J]. 土壤通报, 1986, 17(3):138-141.
	Zhao LP, Jiang Y. The determination method of soil phosphatase activity[J]. Chin J Soil Sci, 1986, 17(3):138-141.
[22]	关松荫. 土壤酶及其研究法[M]. 北京: 农业出版社, 1986:302-305.
	Guan SY. Soil enzymes and research methods[M]. Beijing: Agriculture Press, 1986:302-305.
[23]	Magoč T, Salzberg SL. FLASH:fast length adjustment of short reads to improve genome assemblies[J]. Bioinformatics, 2011, 27(21):2957-2963. doi: 10.1093/bioinformatics/btr507 URL
[24]	Bolger AM, Lohse M, Usadel B. Trimmomatic:a flexible trimmer for Illumina sequence data[J]. Bioinformatics, 2014, 30(15):2114-2120. doi: 10.1093/bioinformatics/btu170 URL
[25]	Edgar RC, Haas BJ, Clemente JC, et al. UCHIME improves sensitivity and speed of chimera detection[J]. Bioinformatics, 2011, 27(16):2194-2200. doi: 10.1093/bioinformatics/btr381 URL
[26]	Edgar RC. UPARSE:highly accurate OTU sequences from microbial amplicon reads[J]. Nat Methods, 2013, 10(10):996-998. doi: 10.1038/nmeth.2604 URL
[27]	Schloss PD, Westcott SL, Ryabin T, et al. Introducing mothur:open-source, platform-independent, community-supported software for describing and comparing microbial communities[J]. Appl Environ Microbiol, 2009, 75(23):7537-7541. doi: 10.1128/AEM.01541-09 URL
[28]	Oksanen J, Blanchet FG, Friendly M, et al. vegan：Community Ecology Package[J]. R package version 2, 2018. https://CRAN.R-project.org/package=vegan
[29]	R Core Team. R：A language and environment for statistical computing[J]. R foundation for statistical computing, Vienna, Austria2017. URL https://www.R-project.org/ .
[30]	Fierer N. Embracing the unknown:disentangling the complexities of the soil microbiome[J]. Nat Rev Microbiol, 2017, 15(10):579-590. doi: 10.1038/nrmicro.2017.87 pmid: 28824177
[31]	Jacoby R, Peukert M, Succurro A, et al. The role of soil microorganisms in plant mineral nutrition-current knowledge and future directions[J]. Front Plant Sci, 2017, 8:1617. doi: 10.3389/fpls.2017.01617 URL
[32]	Wang HH, Li X, Li X, et al. Long-term no-tillage and different residue amounts alter soil microbial community composition and increase the risk of maize root rot in northeast China[J]. Soil Tillage Res, 2020, 196:104452. doi: 10.1016/j.still.2019.104452 URL
[33]	Feng YX, Hu YY, Wu JS, et al. Change in microbial communities, soil enzyme and metabolic activity in a Torreya grandis plantation in response to root rot disease[J]. For Ecol Manag, 2019, 432:932-941. doi: 10.1016/j.foreco.2018.10.028 URL
[34]	Dong L, Xu J, Feng G, et al. Soil bacterial and fungal community dynamics in relation to Panax notoginseng death rate in a continuous cropping system[J]. Sci Rep, 2016, 6:31802. doi: 10.1038/srep31802 URL
[35]	Zhao YP, Wu LK, Chu LX, et al. Interaction of Pseudostellaria heterophylla with Fusarium oxysporum f. sp. heterophylla mediated by its root exudates in a consecutive monoculture system[J]. Sci Rep, 2015, 5(1):1-7.
[36]	Arzanlou M, Torbati M, Golmohammadi H. Powdery mildew on hazelnut(Corylus avellana)caused by Erysiphe corylacearum in Iran[J]. For Path, 2018, 48(5):e12450. doi: 10.1111/efp.2018.48.issue-5 URL
[37]	Cho SE, Lee SH, et al. Erysiphe alphitoides causes powdery mildew on Eucalyptus gunnii[J]. For Path, 2018, 48(1):e12377. doi: 10.1111/efp.2018.48.issue-1 URL
[38]	Singh VK, Singh M, Singh SK, et al. Sustainable agricultural practices using beneficial fungi under changing climate scenario[M]// Climate Change and Agricultural Ecosystems. Amsterdam:Elsevier, 2019:25-42.
[39]	Al-Badi RS, Karunasinghe TG, Al-Sadi AM, et al. In vitro antagonistic activity of endophytic fungi isolated from shirazi thyme(Zataria multiflora boiss. )against Monosporascus cannonballus[J]. Pol J Microbiol, 2020, 69:1-5. doi: 10.33073/pjm-2020-003 pmid: 32067440
[40]	Anandham R, Premalatha N, Jee HJ, et al. Cultivable bacterial diversity and early plant growth promotion by the traditional organic formulations prepared using organic waste materials[J]. Int J Recycl Org Waste Agric, 2015, 4(4):279-289. doi: 10.1007/s40093-015-0107-1 URL
[41]	Shen Z, Penton CR, Lv N, et al. Banana Fusarium wilt disease incidence is influenced by shifts of soil microbial communities under different monoculture spans[J]. Microb Ecol, 2018, 75(3):739-750. doi: 10.1007/s00248-017-1052-5 URL
[42]	Liu N, Shao C, Sun H, et al. Arbuscular mycorrhizal fungi biofertilizer improves American ginseng(Panax quinquefolius L. )growth under the continuous cropping regime[J]. Geoderma, 2020, 363:114155. doi: 10.1016/j.geoderma.2019.114155 URL
[43]	Zhang J, Wei L, Yang J, et al. Probiotic consortia:reshaping the rhizospheric microbiome and its role in suppressing root-rot disease of Panax notoginseng[J]. Front Microbiol, 2020, 11:701. doi: 10.3389/fmicb.2020.00701 URL
[44]	张家春, 孙超, 李朝桢, 等. 不同种植年限白及土壤有机质、酶活性与白及有效成分研究[J]. 中药材, 2020, 43(1):1-4.
	Zhang JC, Sun C, Li CZ, et al. Study on soil organic matter, enzyme activity and the effective components of Bletilla striata under different planting years[J]. J Chin Med Mater, 2020, 43(1):1-4.
[45]	宋旭红, 王钰, 李隆云, 等. 石柱黄连根腐病根际土壤细菌微生态研究[J]. 中国中药杂志, 2017, 42(7):1304-1311.
	Song XH, Wang Y, Li LY, et al. Research on bacteria microecology in root rot rhizosphere soil of Coptis chinensis produced in Shizhu city[J]. China J Chin Mater Med, 2017, 42(7):1304-1311.
[46]	Liu LL, Huang XQ, Zhang JB, et al. Deciphering the relative importance of soil and plant traits on the development of rhizosphere microbial communities[J]. Soil Biol Biochem, 2020, 148:107909. doi: 10.1016/j.soilbio.2020.107909 URL
[47]	刘东海, 张学江, 王鹏, 等. 不同施肥处理对小麦根际土壤真菌多样性及根腐病的影响[J]. 湖北农业科学, 2020, 59(21):30-34, 50.
	Liu DH, Zhang XJ, Wang P, et al. Effects of different fertilization treatments on wheat root rot and the diversity of fungi in rhizosphere soil[J]. Hubei Agric Sci, 2020, 59(21):30-34, 50.
[48]	Zhao LY, Guan HL, Wang R, et al. Effects of tobacco stem-derived biochar on soil properties and bacterial community structure under continuous cropping of Bletilla striata[J]. J Soil Sci Plant Nutr, 2021, 21(2):1318-1328. doi: 10.1007/s42729-021-00442-y URL

真菌Fungi			细菌Bacteria
Phylum	D	H	Phylum	D	H
Ascomycota	73.43%±6.59%^a	40.92%±10.96%^b	Proteobacteria	24.71%±1.62%^a	28.36%±1.76%^a
Mortierellomycota	11.78%±4.19%^b	42.50%±9.86%^a	Bacteroidetes	28.24%±2.35%^a	22.57%±2.90%^a
Basidiomycota	6.90%±0.90^a	12.08±7.47%^a	Acidobacteria	12.78%±1.458%^a	15.85%±2.525%^a
Chytridiomycota	0.39%±0.20%^a	0.12%±0.078%^a	Firmicutes	13.68%±1.62%^a	9.05%±1.82%^a
Rozellomycota	0.10%±0.042%^a	0.028%±0.023%^a	Actinobacteria	7.71%±0.86%^a	8.29%±0.98%^a
Olpidiomycota	0.083%±0.054%^a	0.012%±0.009%^a	Gemmatimonadetes	5.25%±0.49%^a	6.42%±0.61%^a
Others	7.32%±1.95%^a	4.34%±0.69%^a	Others	7.64%±0.29%^a	9.47%±0.79%^a

真菌Fungi			细菌Bacteria
Phylum	D	H	Phylum	D	H
Ascomycota	73.43%±6.59%^a	40.92%±10.96%^b	Proteobacteria	24.71%±1.62%^a	28.36%±1.76%^a
Mortierellomycota	11.78%±4.19%^b	42.50%±9.86%^a	Bacteroidetes	28.24%±2.35%^a	22.57%±2.90%^a
Basidiomycota	6.90%±0.90^a	12.08±7.47%^a	Acidobacteria	12.78%±1.458%^a	15.85%±2.525%^a
Chytridiomycota	0.39%±0.20%^a	0.12%±0.078%^a	Firmicutes	13.68%±1.62%^a	9.05%±1.82%^a
Rozellomycota	0.10%±0.042%^a	0.028%±0.023%^a	Actinobacteria	7.71%±0.86%^a	8.29%±0.98%^a
Olpidiomycota	0.083%±0.054%^a	0.012%±0.009%^a	Gemmatimonadetes	5.25%±0.49%^a	6.42%±0.61%^a
Others	7.32%±1.95%^a	4.34%±0.69%^a	Others	7.64%±0.29%^a	9.47%±0.79%^a

分组 Group	真菌Fungi			细菌Bacteria
分组 Group	OTU richness	Chao1	Shannon	OTU richness	Chao1	Shannon
D	639.8±40.30^a	675.09±45.02^a	3.82±0.58^a	1 607.20±11.03^a	1 647.26±11.19^a	6.45±0.02^a
H	526.0±6.31^b	639.43±38.54^a	2.92±0.33^a	1 619.00±13.17^a	1 668.55±8.14^a	6.37±0.04^a

分组 Group	真菌Fungi			细菌Bacteria
分组 Group	OTU richness	Chao1	Shannon	OTU richness	Chao1	Shannon
D	639.8±40.30^a	675.09±45.02^a	3.82±0.58^a	1 607.20±11.03^a	1 647.26±11.19^a	6.45±0.02^a
H	526.0±6.31^b	639.43±38.54^a	2.92±0.33^a	1 619.00±13.17^a	1 668.55±8.14^a	6.37±0.04^a

土壤因子Soil factor	H	D	P
pH	6.57±0.19	6.87±0.18	<0.05
EC（μS·cm^-1）	82.06±5.25	65.80±4.14	<0.05
铵态氮（mg·kg^-1）	4.35±0.42	5.12±0.36	<0.05
硝态氮（mg·kg^-1）	3.02±0.35	6.49±0.60	<0.01
有机质（g·kg^-1）	4.84±0.44	5.68±0.52	<0.05
有效磷（mg·kg^-1）	81.36±17.01	77.05±28.64	0.78
有效钾（mg·kg^-1）	414.12±14.71	358.00±15.81	<0.05
脲酶（mg·NH₄⁺-N·g^-1 24 h^-1）	0.51±0.03	0.46±0.03	<0.05
磷酸酶（mg phenol·g^-1·12 h^-1）	0.14±0.01	0.13±0.01	0.13
蛋白酶（mg·NH₂-N·g^-1·24 h^-1）	0.044±0.01	0.036±0.01	<0.01
蔗糖酶（mg glucose·g^-1·24 h^-1）	22.49±0.89	15.90±0.80	<0.01