Characteristics of the Mycosphere Microbial Community in Diseased and Healthy Morchella spp.Soil

doi:10.13560/j.cnki.biotech.bull.1985.2024-0819

Abstract

Abstract:

Objective To explore the microbial characteristics in the Morchella spp. soil and provide scientific guidance for the rational cultivation of Morchella spp. Method High-throughput sequencing technology was applied to determine the communities and diversity indices of bacteria and fungi in the soils growing diseased and healthy Morchella spp., and further analyze their roles for transformation of soil substances, nutrient uptake and disease resistances of plant. Result The contents of available phosphorus, pH and the activity of catalase in the diseased Morchella spp. soil were significantly higher than those in healthy Morchella spp. soil, increasing with 13.22%, 6.18%, and 41.01%, respectively, while the activities of urease, acid phosphatase and sucrase significantly decreased with 58.58%, 31.85%, and 74.01% respectively. The bacterial indices of Chao1, ACE and Simpson in DM significantly decreased with 32.45%, 32.43%, and 20.42%, respectively compared to those in HM, and there was no significant change for fungal indices. The results of principal component analysis indicated that the community structure of bacteria (R²=0.195, P=0.028) and fungi (R²=0.17, P=0.001) in DM were significantly different from that in HM. The abundance of beneficial bacteria of Haliangium, Gemmatimonas and Flavobacterium in DMwere significantly lower than that in HM, which decreased by 92.85%, 90.48%, and 81.67%. The abundance of Enterobacter and Polaromonas in DMwere significantly higher than that in HM, which increased with 40.33 and 8.13 times, respectively. The abundance of beneficial fungus Schizothecium in DMwas significantly lower than that in HM, decreasing by 90.39%, and the abundance of Fusarium in the DM were significantly higher than that in HM, increasing with 2.51 times. The results of functional prediction showed that the metabolic pathways of bacteria in diseased Morchella spp. soil were different with that in healthy Morchella spp. soil, and the metabolic type of fungi was mainly saprotrophic. The results of Mantel analysis showed that the pH was the key factor affecting the fungal microbial community. The analysis of co-occurrence network showed that the average degree, total module number and average clustering coefficient of the bacterial and fungal network in the diseased Morchella spp. group were lower than those in the healthy group. Conclusion The richness and diversity of soil bacteria in the diseased Morchella spp. significantly decrease, the diseased Morchella spp. soil abundance of pathogen increase and that of the beneficial microbe decrease, and the complexity and stability of microbial network reduce, which are the important factors for the diseased Morchella spp.

Key words: Morchella spp., mycosphere soil, composition of microbial community, white mold disease, structure of microbial community, soil enzyme activity, soil properties

SONG Fen-fen, DUAN Yan-xue, SANG Yu, WANG Ji-peng, PENG Rui, SUN Nian-xi, LI Yong. Characteristics of the Mycosphere Microbial Community in Diseased and Healthy Morchella spp.Soil[J]. Biotechnology Bulletin, 2025, 41(4): 323-334.

Figures/Tables 13

References 52

11	刘建明, 冯吉昌, 吴之涛, 等. 河西地区羊肚菌白霉病病原鉴定及生物学特性研究 [J]. 北方园艺, 2023(17): 116-123.
	Liu JM, Feng JC, Wu ZT, et al. Study on pathogen identification and biological morel of Morchella white mold disease in Hexi area [J]. North Hortic, 2023(17): 116-123.
12	Sun JZ, Yu S, Lu YZ, et al. Proposal of a new family Pseudodiploösporeaceae fam. nov. (Hypocreales) based on phylogeny of Diploöspora longispora and Paecilomyces penicillatus [J]. Mycology, 2022, 14(1): 60-73.
13	Masaphy S. First report on Purpureocillium lilacinum infection of indoor-cultivated morel primordia [J]. Agriculture, 2022, 12(5): 695.
14	Zhu XT, Ma KL, Sun MY, et al. Isolation and identification of pathogens of Morchella sextelata bacterial disease [J]. Front Microbiol, 2023, 14: 1231353.
15	Yu FM, Jayawardena RS, Luangharn T, et al. Species diversity of fungal pathogens on cultivated mushrooms: a case study on morels (Morchella, Pezizales) [J]. Fungal Divers, 2024, 125(1): 157-220.
16	He XL, Peng WH, Miao RY, et al. White mold on cultivated morels caused by Paecilomyces penicillatus [J]. FEMS Microbiol Lett, 2017, 364(5). DOI:10.1093/femsle/fnx037 .
17	余苗, 尹琪, 何培新. 羊肚菌白腐病病原菌的分离与鉴定 [J]. 北方园艺, 2020(7): 142-145.
	Yu M, Yin Q, He PX. Isolation and identification of pathogen of morel white rot [J]. North Hortic, 2020(7): 142-145.
18	陈诚, 李强, 黄文丽, 等. 羊肚菌白霉病发生对土壤真菌群落结构的影响 [J]. 微生物学通报, 2017, 44(11): 2652-2659.
	Chen C, Li Q, Huang WL, et al. Effects of Morchella white mold disease on soil fungal community structure [J]. Microbiol China, 2017, 44(11): 2652-2659.
19	孟庆国, 赵英同. 栽培羊肚菌常见病虫害及预防措施 [J]. 食用菌, 2021, 43(6): 66-67.
	Meng QG, Zhao YT. Common pests and diseases of cultivated Morchella and their preventive measures [J]. Edible Fungi, 2021, 43(6): 66-67.
20	米其利, 李雪梅, 管莹, 等. 高通量测序在食品微生物生态学研究中的应用 [J]. 食品科学, 2016, 37(23): 302-308.
1	杜习慧, 赵琪, 杨祝良. 羊肚菌的多样性、演化历史及栽培研究进展 [J]. 菌物学报, 2014, 33(2): 183-197.
	Du XH, Zhao Q, Yang ZL. Diversity, evolutionary history and cultivation of morels: a review [J]. Mycosystema, 2014, 33(2): 183-197.
2	熊川, 黄文丽, 金鑫, 等. 一种羊肚菌源多肽MIP-16的分离纯化及其神经保护活性 [J]. 天然产物研究与开发, 2021, 33(9): 1519-1526.
	Xiong C, Huang WL, Jin X, et al. Isolation and purification of a novel peptide from the fruiting bodies of Morchella importuna and its neuroprotective effect [J]. Nat Prod Res Dev, 2021, 33(9): 1519-1526.
3	Wang DD, Yin ZQ, Ma LK, et al. Polysaccharide MCP extracted from Morchella esculenta reduces atherosclerosis in LDLR-deficient mice [J]. Food Funct, 2021, 12(11): 4842-4854.
4	Ramya H, Ravikumar KS, Fathimathu Z, et al. Morel mushroom, Morchella from Kashmir Himalaya: a potential source of therapeutically useful bioactives that possess free radical scavenging, anti-inflammatory, and arthritic edema-inhibiting activities [J]. Drug Chem Toxicol, 2022, 45(5): 2014-2023.
5	Du XH, Wu DM, He GQ, et al. Six new species and two new records of Morchella in China using phylogenetic and morphological analyses [J]. Mycologia, 2019, 111(5): 857-870.
6	武冬梅, 李先义, 高能, 等. 采用多基因联合方法鉴定新疆野生羊肚菌 [J]. 分子植物育种, 2022, 20(7): 2420-2427.
	Wu DM, Li XY, Gao N, et al. Taxonomic identification of wild Morchella in Xinjiang based on multigene [J]. Mol Plant Breed, 2022, 20(7): 2420-2427.
7	蔡英丽, 马晓龙, 路等学, 等. 野生羊肚菌Mel-21的系统发育分析和驯化 [J]. 食用菌学报, 2020, 27(3): 23-29.
	Cai YL, Ma XL, Lu DX, et al. Phylogenetic analysis and domestication of wild morel Mel -21 [J]. Acta Edulis Fungi, 2020, 27(3): 23-29.
8	李建英, 华蓉, 孙达锋, 等. 羊肚菌白霉病病原菌及病害样地土壤真菌群落结构研究 [J]. 中国食用菌, 2022, 41(10): 50-54.
20	Mi QL, Li XM, Guan Y, et al. Application of high-throughput sequencing in food microbial ecology: a review [J]. Food Sci, 2016, 37(23): 302-308.
21	卓娜, 伊丽, 浩斯娜, 等. 基于16S rRNA基因序列分析法比较苏尼特双峰驼和阿拉善双峰驼自然发酵酸驼乳的微生物多样性 [J]. 微生物学报, 2019, 59(10): 1948-1959.
	Zhuo N, Yi L, Hao SN, et al. Application of 16S rRNA high-throughput sequencing for comparative study of the microbial diversity of traditional fermented Bactrian camel milk from Alxa Bactrian camel and Sonid Bactrian camel [J]. Acta Microbiol Sin, 2019, 59(10): 1948-1959.
22	吴振强, 戴瑞卿, 赖宝春, 等. 健康与感染枯萎病辣椒植株根际土壤真菌群落结构与多样性 [J]. 现代农业科技, 2020(7): 184-187.
	Wu ZQ, Dai RQ, Lai BC, et al. Fungus community structure and diversity of rhizosphere soil of healthy and Fusarium wilt chilli plants [J]. Mod Agric Sci Technol, 2020(7): 184-187.
23	Santhanam R, Luu VT, Weinhold A, et al. Native root-associated bacteria rescue a plant from a sudden-wilt disease that emerged during continuous cropping [J]. Proc Natl Acad Sci U S A, 2015, 112(36): E5013-E5020.
24	Bakker PAHM, Doornbos RF, Zamioudis C, et al. Induced systemic resistance and the rhizosphere microbiome [J]. Plant Pathol J, 2013, 29(2): 136-143.
25	彭博, 边银丙, 龚钰华, 等. 大田栽培羊肚菌的病害及其综合防控技术 [J]. 食药用菌, 2024, 32(2): 129-135.
	Peng B, Bian YB, Gong YH, et al. Diseases of Morchella and their comprehensive prevention and control techniques [J]. Edible Med Mushrooms, 2024, 32(2): 129-135.
26	鲍士旦. 土壤农化分析 [M]. 3版. 北京: 中国农业出版社, 2000.
	Bao SD. Soil and agricultural chemistry analysis [M]. 3rd ed. Beijing: China Agriculture Press, 2000.
27	关松荫. 土壤酶及其研究法 [M]. 北京: 农业出版社, 1986.
8	Li JY, Hua R, Sun DF, et al. Research on white mold disease on Morchella spp. and fungal community structure of soil samples [J]. Edible Fungi China, 2022, 41(10): 50-54.
9	苏文英, 刘晓梅, 纪伟, 等. 羊肚菌白霉病病原鉴定及生物学特性研究 [J]. 浙江农业科学, 2023, 64(1): 204-208.
	Su WY, Liu XM, Ji W, et al. Pathogen identification and biological characteristics of Morchella esculenta [J]. J Zhejiang Agric Sci, 2023, 64(1): 204-208.
10	Dong W, Chen BS, Zhang R, et al. Identification and characterization of peptaibols as the causing agents of Pseudodiploöspora longispora infecting the edible mushroom Morchella [J]. J Agric Food Chem, 2023, 71(47): 18385-18394.
27	Guan SY. Soil enzyme and its research method [M]. Beijing: Agriculture Press, 1986.
28	Li JR, Chen XZ, Li SM, et al. Variations of rhizospheric soil microbial communities in response to continuous Andrographis paniculata cropping practices [J]. Bot Stud, 2020, 61(1): 18.
29	Huang WJ, Sun DL, Fu JT, et al. Effects of continuous sugar beet cropping on rhizospheric microbial communities [J]. Genes, 2019, 11(1): 13.
30	Edgar RC. UPARSE: highly accurate OTU sequences from microbial amplicon reads [J]. Nat Methods, 2013, 10(10): 996-998.
31	Bokulich NA, Subramanian S, Faith JJ, et al. Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing [J]. Nat Methods, 2013, 10(1): 57-59.
32	Wang Q, Garrity GM, Tiedje JM, et al. Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy [J]. Appl Environ Microbiol, 2007, 73(16): 5261-5267.
33	Quast C, Pruesse E, Yilmaz P, et al. The SILVA ribosomal RNA gene database project: improved data processing and web-based tools [J]. Nucleic Acids Res, 2013, 41(Database issue): D590-D596.
34	Kõljalg U, Henrik Nilsson R, Abarenkov K, et al. Towards a unified paradigm for sequence-based identification of fungi [J]. Mol Ecol, 2013, 22(21): 5271-5277.
35	姚丽娟, 田春丽, 王立河, 等. 设施西瓜连作土壤生化性质及微生物群落变化 [J]. 中国土壤与肥料, 2024(3): 70-78.
	Yao LJ, Tian CL, Wang LH, et al. Changes of soil biochemical properties and microbial community in continuous cropping of watermelon under greenhouse cultivation [J]. Soils Fertil Sci China, 2024(3): 70-78.
36	Houfani AA, Větrovský T, Navarrete OU, et al. Cellulase-hemicellulase activities and bacterial community composition of different soils from Algerian ecosystems [J]. Microb Ecol, 2019, 77(3): 713-725.
37	黄海莉, 宗宁, 何念鹏, 等. 青藏高原高寒草甸不同海拔土壤酶化学计量特征 [J]. 应用生态学报, 2019, 30(11): 3689-3696.
	Huang HL, Zong N, He NP, et al. Characteristics of soil enzyme stoichiometry along an altitude gradient on Qinghai-Tibet Pla-teau alpine meadow, China [J]. Chin J Appl Ecol, 2019, 30(11): 3689-3696.
38	赵娟, 贾卫国, 刘伟成, 等. 草莓灰霉病菌拮抗放线菌的筛选及活性测定 [J]. 北方园艺, 2018(4): 59-65.
	Zhao J, Jia WG, Liu WC, et al. Screening of antagonistic actinomycete against strawberry grey mould and activity determination of its fermentation broth [J]. North Hortic, 2018(4): 59-65.
39	边雪廉, 岳中辉, 焦浩, 等. 土壤酶对土壤环境质量指示作用的研究进展 [J]. 土壤, 2015, 47(4): 634-640.
	Bian XL, Yue ZH, Jiao H, et al. Soil enzyme indication on soil environmental quality: a review [J]. Soils, 2015, 47(4): 634-640.
40	林睿. 土壤链霉菌拮抗致病疫霉及其对植物促生特性的研究 [D]. 呼和浩特: 内蒙古农业大学, 2019.
	Lin R. Antagonism of Streptomyces terrestris against Phytophthora infestans and its characteristics of promoting plant growth [D]. Hohhot: Inner Mongolia Agricultural University, 2019.
41	何国兴, 宋建超, 温雅洁, 等. 不同根瘤菌肥对紫花苜蓿生产力及土壤肥力的综合影响 [J]. 草业学报, 2020, 29(5): 109-120.
	He GX, Song JC, Wen YJ, et al. Effects of different Rhizobium fertilizers on alfalfa productivity and soil fertility [J]. Acta Prataculturae Sin, 2020, 29(5): 109-120.
42	Tian XL, Wang DD, Mao ZC, et al. Infection of Plasmodiophora brassicae changes the fungal endophyte community of tumourous stem mustard roots as revealed by high-throughput sequencing and culture-dependent methods [J]. PLoS One, 2019, 14(6): e0214975.
43	刘海洋, 姚举, 张仁福, 等. 黄萎病不同发生程度棉田中土壤微生物多样性 [J]. 生态学报, 2018, 38(5): 1619-1629.
	Liu HY, Yao J, Zhang RF, et al. Analysis of soil microbial diversity in cotton fields differing in occurrence of cotton Verticillium wilt in Xinjiang [J]. Acta Ecol Sin, 2018, 38(5): 1619-1629.
44	张丽娟, 曲继松, 郭文忠, 等. 微生物菌肥对黄河上游地区设施土壤微生物及酶活性的影响 [J]. 中国土壤与肥料, 2014(5): 32-36, 99.
	Zhang LJ, Qu JS, Guo WZ, et al. Effects of the microbial fertilizers on microorganism and enzymic activity in greenhouse soil on upper reaches of the Yellow River [J]. Soil Fertil Sci China, 2014(5): 32-36, 99.
45	张雪, 张学学, 胡鑫, 等. 皂角发酵物对红托竹荪病害防治及土壤主要微生物群落影响 [J]. 菌物学报, 2022, 41(4): 618-629.
	Zhang X, Zhang XX, Hu X, et al. Effects of Gleditsia sinensis fermentation product on disease control and main soil microbial community of Dictyophora rubrovolvata [J]. Mycosystema, 2022, 41(4): 618-629.
46	姜艳鹏, 胡振华, 张翠静, 等. 枯草芽孢杆菌J22拮抗姜瘟阴沟肠杆菌的效果与机理研究 [J]. 江苏农业科学, 2023, 51(2): 125-134.
	Jiang YP, Hu ZH, Zhang CJ, et al. Antagonistic effect and mechanism of Bacillus subtilis J22 against Enterobacter cloacae in ginger blast [J]. Jiangsu Agric Sci, 2023, 51(2): 125-134.
47	Ujvári G, Turrini A, Avio L, et al. Possible role of arbuscular mycorrhizal fungi and associated bacteria in the recruitment of endophytic bacterial communities by plant roots [J]. Mycorrhiza, 2021, 31(5): 527-544.
48	Rhee SY, Mutwil M. Towards revealing the functions of all genes in plants [J]. Trends Plant Sci, 2014, 19(4): 212-221.
49	Mondo SJ, Jiménez DJ, Hector RE, et al. Genome expansion by allopolyploidization in the fungal strain Coniochaeta 2T2.1 and its exceptional lignocellulolytic machinery [J]. Biotechnol Biofuels, 2019, 12: 229.
50	黄慧, 张晓勇, 郑欢, 等. 羊肚菌菌盖干腐病病原菌鉴定及培养特性研究 [J]. 植物保护, 2022, 48(1): 66-72.
	Huang H, Zhang XY, Zheng H, et al. Identification and cultural characterization of Diploöspora longispora associated with Pileus rot disease on cultivated morel [J]. Plant Prot, 2022, 48(1): 66-72.
51	韩梦颖, 王雨桐, 高丽, 等. 降解秸秆微生物及秸秆腐熟剂的研究进展 [J]. 南方农业学报, 2017, 48(6): 1024-1030.
	Han MY, Wang YT, Gao L, et al. Straw degradation microorganism and straw-decomposing inoculant: a review [J]. J South Agric, 2017, 48(6): 1024-1030.
52	Mahdi LK, Miyauchi S, Uhlmann C, et al. The fungal root endophyte Serendipita vermifera displays inter-Kingdom synergistic beneficial effects with the microbiota in Arabidopsis thaliana and barley [J]. ISME J, 2022, 16(3): 876-889.

微生物 Microorganism	组别 Group	Chao1指数 Chao1 index	ACE指数 ACE index	Shannon指数 Shannon index	覆盖度 Coverage/%
细菌 Bacteria	HM	517.38±17.41a	517.40±17.48a	8.62±0.10a	100.00
细菌 Bacteria	DM	349.50±48.77b	349.59±48.79b	6.86±0.36b	99.98
真菌 Fungi	HM	154.29±15.78a	158.76±17.02a	3.03±1.32a	99.98
真菌 Fungi	DM	133.04±29.90a	133.00±28.74a	2.43±0.74a	99.99

微生物 Microorganism	组别 Group	Chao1指数 Chao1 index	ACE指数 ACE index	Shannon指数 Shannon index	覆盖度 Coverage/%
细菌 Bacteria	HM	517.38±17.41a	517.40±17.48a	8.62±0.10a	100.00
细菌 Bacteria	DM	349.50±48.77b	349.59±48.79b	6.86±0.36b	99.98
真菌 Fungi	HM	154.29±15.78a	158.76±17.02a	3.03±1.32a	99.98
真菌 Fungi	DM	133.04±29.90a	133.00±28.74a	2.43±0.74a	99.99

微生物类别 Microbial type	分类 Taxonomy	健康羊肚菌 HM/%	患病羊肚菌DM/%
细菌 Bacteria	Acidibacter	0.87±0.56a	0.11±0.05b
	Biyi10	1.13±0.85a	0.09±0.24b
	Bosea	0.09±0.08b	0.44±0.12a
	Candidatus_Koribacter	0.46±0.34a	0.07±0.16b
	Dongia	1.88±0.97a	0.47±0.74b
	Ellin6067	0.86±0.64a	0.08±0.24b
	Enterobacter	0.02±0.04b	0.87±0.15a
	Flavisolibacter	0.60±0.36a	0.11±0.07b
	Flavitalea	0.30±0.07a	0.11±0.11b
	Gemmatimonas	1.65±0.01a	0.157±1.23b
	Haliangium	0.91±0.29a	0.07±0.30b
	IS_44	0.57±0.42a	0.01±0.05b
	Lautropia	0.26±0.25a	0.03±0.02b
	Luteitalea	0.87±0.54a	0.19±0.26b
	Methylobacillus	0.33±0.26a	0.04±0.04b
	Nannocystis	0.15±0.10a	0.02±0.07b
	OLB13	0.49±0.29a	0.02±0.37b
	Opitutus	0.13±0.12a	0.01±0.02b
	Pajaroellobacter	0.09±0.03a	0.00±0.03b
	Polaromonas	0.15±0.14b	1.22±0.74a
	Stella	0.17±0.07a	0.00±0.04b
	Stenotrophomonas	0.15±0.12b	3.54±0.39a
	Thalassobaculum	0.13±0.08a	0.03±0.04b
	UTBCD1	0.43±0.43a	0.00±0.00b
	Variovorax	0.28±0.27b	1.37±0.26a
真菌 Fungi	Aspergillus	0.69±0.48a	0.08±0.04b
	Fusarium	1.00±0.69b	3.50±1.92a
	Schizothecium	4.01±3.03a	0.39±0.50b
	Serendipita	0.00±0.00a	0.00±0.00b

微生物类别 Microbial type	分类 Taxonomy	健康羊肚菌 HM/%	患病羊肚菌DM/%
细菌 Bacteria	Acidibacter	0.87±0.56a	0.11±0.05b
	Biyi10	1.13±0.85a	0.09±0.24b
	Bosea	0.09±0.08b	0.44±0.12a
	Candidatus_Koribacter	0.46±0.34a	0.07±0.16b
	Dongia	1.88±0.97a	0.47±0.74b
	Ellin6067	0.86±0.64a	0.08±0.24b
	Enterobacter	0.02±0.04b	0.87±0.15a
	Flavisolibacter	0.60±0.36a	0.11±0.07b
	Flavitalea	0.30±0.07a	0.11±0.11b
	Gemmatimonas	1.65±0.01a	0.157±1.23b
	Haliangium	0.91±0.29a	0.07±0.30b
	IS_44	0.57±0.42a	0.01±0.05b
	Lautropia	0.26±0.25a	0.03±0.02b
	Luteitalea	0.87±0.54a	0.19±0.26b
	Methylobacillus	0.33±0.26a	0.04±0.04b
	Nannocystis	0.15±0.10a	0.02±0.07b
	OLB13	0.49±0.29a	0.02±0.37b
	Opitutus	0.13±0.12a	0.01±0.02b
	Pajaroellobacter	0.09±0.03a	0.00±0.03b
	Polaromonas	0.15±0.14b	1.22±0.74a
	Stella	0.17±0.07a	0.00±0.04b
	Stenotrophomonas	0.15±0.12b	3.54±0.39a
	Thalassobaculum	0.13±0.08a	0.03±0.04b
	UTBCD1	0.43±0.43a	0.00±0.00b
	Variovorax	0.28±0.27b	1.37±0.26a
真菌 Fungi	Aspergillus	0.69±0.48a	0.08±0.04b
	Fusarium	1.00±0.69b	3.50±1.92a
	Schizothecium	4.01±3.03a	0.39±0.50b
	Serendipita	0.00±0.00a	0.00±0.00b

网络性质 Network properties	细菌 Bacteria		真菌 Fungi
网络性质 Network properties	健康 HM	患病 DM	健康 HM	患病 DM
平均度Average degree	37.28	35.83	37.28	7.046
模块系数Modularity index	1	0.76	0.65	0.56
总节点Total nodes	562	386	209	65
总边Total edges	10 515	6 530	3 896	164
正连接Positive edges	10 076	6 402	3 881	162
负连接Negative edges	439	128	15	2