柴达木地区患根腐病枸杞根际土壤微生物多样性分析

doi:10.13304/j.nykjdb.2022.0530

摘要/Abstract

摘要：

为了解枸杞受根腐病菌侵染后植株根组织与根际土壤中真菌与细菌群落结构以及多样性，采用高通量测序技术对柴达木地区患根腐病枸杞的根组织和根际土壤样本进行分析。结果表明，根际土壤中的细菌群落丰度略高于根组织，而真菌多样性和丰度在根际土壤和根组织样本中无显著差异。在门水平，细菌群落中变形菌门（Proteobacteria）和拟杆菌门（Bacteroidetes）的相对丰度较高，真菌群落中子囊菌门（Ascomycota）和担子菌门（Basidiomycota）的相对丰度较高；在属水平，细菌中以假单胞菌（Pseudomonas）为主，真菌中以枝孢属（Cladosporium）和被孢霉属（Mortierella）为主。相关性分析显示，细菌群落在根组织和根际土壤样本中差异较大，而真菌群落在根组织和根际土壤样本间相关性较高；枝孢属（Cladosporium）和被孢霉属（Mortierella）的相对丰度在根组织和根际土壤中均显著上调，初步推测这2属真菌可能与枸杞根腐病发病相关。

关键词: 枸杞, 根腐病, 高通量测序, 微生物多样性

Abstract:

To understand the structure and diversity of fungal and bacterial communities in root tissues and rhizosphere soil of Lycium barbarum plants infested with root rot， root tissues and rhizosphere soil samples of Lycium barbarum infested with root rot in Qaidam region were analyzed using high-throughput sequencing technology. The results showed that bacterial species were slightly more abundant in soil than that in root tissue， and fungal diversity and abundance were not significantly different between soil and root tissue samples. At phylum level， Proteobacteria and Bacteroidetes had higher abundance in bacterial communities， and Ascomycota and Basidiomycota had higher abundance in fungal communities. At genus level， Pseudomonas was dominated in bacteria， and Cladosporium and Mortierella were dominated in fungi. The correlation analysis showed that bacteria were more different between root tissue and soil samples， and fungi were better correlated in root tissue and soil samples. Both Cladosporium and Mortierella were significantly upregulated in the root system and soil， which assumed to be associated with the pathogenesis of root rot of Lycium barbarum.

Key words: Lycium barbarum, root rot, high-throughput sequencing, microbial diversity

中图分类号:

S154.3

方泰军, 侯璐, 白露超. 柴达木地区患根腐病枸杞根际土壤微生物多样性分析[J]. 中国农业科技导报, 2024, 26(1): 133-139.

Taijun FANG, Lu HOU, Luchao BAI. Soil Microbial Diversity in the Rhizosphere of Lycium barbarum Infected with Root Rot Disease in the Qaidam Region[J]. Journal of Agricultural Science and Technology, 2024, 26(1): 133-139.

图/表 8

参考文献 20

1	YAO R, HEINRICH M, ZOU Y, et al.. Quality variation of gouji (fruits of Lycium spp.) in China: a comparative morphological and metabolomic analysis [J/OL]. Front Pharmacol., 2018, 9:151 [2022-05-10]. .
2	CHEN C, SHAO Y, LI Y, et al.. Trace elements in Lycium barbarum L. leaves by inductively coupled plasma mass spectrometry after microwave assisted digestion and multivariate analysis [J]. Spectrosc. Lett., 2015, 48:775-780.
3	王凤宝,付金锋,董立峰,等.多倍体菜用枸杞新品种天精 3 号选育[J].核农学报,2013,27(6):723-730.
	WANG F B, FU J F, DONG L F, et al.. Selection and breeding of the new mixoploid lycium chinense miller variety Tianjing 3 [J]. J. Nucl. Agric. Sci., 2013, 27(6):723-730.
4	CORRADO C, MINCUZZI A, SCHENA L, et al.. First report of collar and root rot caused by Phytophthora nicotianae on Lycium barbarum [J/OL]. J. Plant Pathol., 2018, 100:361 [2022-05-10]. .
5	任如红,胡毅敏,李红叶,等.杨梅根腐病的早期诊断[J].浙江林学院学报,2000(1):117-121.
	REN R H, HU Y M, LI H Y, et al.. Early diagnosis of root rot in Chinese strawberry [J]. J. Zhejiang For. Coll., 2000(1):117-121.
6	WANG L, LI X. Steering soil microbiome to enhance soil system resilience [J]. Crit. Rev. Microbiol., 2019, 45:5-6,743-753.
7	MUNIR N, CHENG C Z, XIA C S, et al.. RNA-Seq analysis reveals an essential role of tyrosine metabolism pathway in response to root-rot infection in Gerbera hybrida [J/OL]. PloS One, 2019, 14(10):223519 [2022-05-10]. .
8	MOREIRA E C, PINHEIRO D G, GORDO S, et al.. Transcriptional profiling by RNA sequencing of black pepper (Piper nigrum L.) roots infected by Fusarium solani f. sp. piperis [J/OL]. Acta Physiol. Plant, 2017, 39(10):239 [2022-05-10]. .
9	MANTIRI F R, SAMUELS G J, RAHE J E, et al.. Phylogenetic relationships in Neonectria species having Cylindrocarpon anamorphs inferred from mitochondrial ribosomal DNA sequences [J]. Can. J. Bot., 2001, 79:334-340.
10	MAVRODI O V, WALTER N, ELATEEK S, et al.. Suppression of rhizoctonia and pythium root rot of wheat by new strains of Pseudomonas [J]. Biol. Control, 2012, 62(2):13-25.
11	TEWOLDEMEDHIN Y T, MAZZOLA M, BOTHA W J, et al.. Characterization of fungi (Fusarium and Rhizoctonia) and oomycetes (Phytophthora and Pythium) associated with apple orchards in South Africa [J]. Eur. J. Plant Pathol., 2011, 130(2):215-229.
12	WU Z, HAO Z, SUN Y, 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.
13	NATACHA B, HORTON M W, BERGELSON J, et al.. Bacterial communities associated with the leaves and the roots of Arabidopsis thaliana [J/OL]. PloS One, 2013, 8:e56329 [2022-05-10]. .
14	CAO Y T, LIU J L. Review on the diseases and insects of Lycium barbarum L. [J]. Plant Dis. Pests, 2014, 5(6):8-12.
15	ROCHON D. Fungal transmission of plant viruses [J]. Annu. Rev. Phytopathol., 2009, 34(1):87-108.
16	LIU Y, SUN Q, LI J, et al.. Bacterial diversity among the fruit bodies of ectomycorrhizal and saprophytic fungi and their corresponding hyphosphere soils [J]. Sci. Rep., 2018, 8(1):1-10.
17	DINLER H, BENLIOGLU S, BENLIOGLU K. Rhizoctonia fragariae causes black root rot on strawberry seedlings in Turkey [J]. Australas. Plant Dis. Notes, 2018, 13(1):1-3.
18	吕靖雯,刘蕊,李国华,等.柚果面枝孢菌斑点病病原鉴定[J].浙江大学学报(农业与生命科学版),2018,44(6):687-694.
	LYU J W, LIU R, LI G H, et al.. Identification of pathogens causing fruit spot disease on Citrus grandis [J]. J. Zhejiang Univ. (Agric. Life Sci.), 2018, 44(6):687-694.
19	雷桂林,刘云龙,刘雪峰,等.雪松针叶枯斑病病原鉴定及其生物学特性研究[J].林业科学研究, 2003(4):453-458.
	LEI G L, LIU Y L, LIU X F, et al.. Determination of necrotic spot pathogen on needle of Cedrus deodara and the biological characteristics of the pathogen [J]. For. Res., 2003(4):453-458.
20	任海英,王程安,俞浙萍,等.杨梅衰弱病根际土壤真菌的定量检测与分析[J].植物病理学报,2022,52(4):529-536.
	REN H Y, WANG C A, YU Z P, et al.. Quantitative detection and analysis of rhizospheric soil fungi in decline disease of Myrica rubra [J]. Acta Phytopathol. Sin., 2022, 52(4):529-536.

样品编号 Sample ID	双端reads PE reads	原始序列 Raw tag	优化序列 Clean tag	有效序列 Effective tag	平均长度 Avglen/bp	GC/%	Q30/%	有效性 Effective/%
S1	138 258	125 656	119 310	119 145	261	45.60	97.24	86.18
S2	230 942	184 074	174 566	174 494	249	48.44	97.26	75.56
S3	170 588	151 295	142 645	142 567	287	59.33	96.13	83.57
S4	161 081	142 320	135 436	135 320	249	50.22	97.25	84.01
S6	169 066	149 480	142 190	142 016	247	53.14	97.28	84.00
R1	168 773	144 072	136 387	136 218	268	53.10	96.67	80.71
R2	130 623	117 505	104 076	104 001	252	51.96	97.20	79.62
R3	91 298	80 917	76 857	76 738	248	47.16	97.38	84.05
R4	116 581	104 969	100 082	99 942	235	49.92	97.77	85.73
R5	117 686	107 380	102 557	102 520	232	48.68	98.12	87.11
R6	90 044	80 035	76 309	76 237	242	45.97	97.41	84.67

样品编号 Sample ID	双端reads PE reads	原始序列 Raw tag	优化序列 Clean tag	有效序列 Effective tag	平均长度 Avglen/bp	GC/%	Q30/%	有效性 Effective/%
S1	138 258	125 656	119 310	119 145	261	45.60	97.24	86.18
S2	230 942	184 074	174 566	174 494	249	48.44	97.26	75.56
S3	170 588	151 295	142 645	142 567	287	59.33	96.13	83.57
S4	161 081	142 320	135 436	135 320	249	50.22	97.25	84.01
S6	169 066	149 480	142 190	142 016	247	53.14	97.28	84.00
R1	168 773	144 072	136 387	136 218	268	53.10	96.67	80.71
R2	130 623	117 505	104 076	104 001	252	51.96	97.20	79.62
R3	91 298	80 917	76 857	76 738	248	47.16	97.38	84.05
R4	116 581	104 969	100 082	99 942	235	49.92	97.77	85.73
R5	117 686	107 380	102 557	102 520	232	48.68	98.12	87.11
R6	90 044	80 035	76 309	76 237	242	45.97	97.41	84.67

样品名称 Sample ID	双端reads PE reads	原始序列 Raw tag	优化序列 Clean tag	有效序列 Effective tag	平均长度 Avglen/bp	GC/%	Q30/%	有效性 Effective/%
S1	80 094	75 613	67 304	65 710	420	56.36	93.52	82.04
S2	79 987	75 747	67 761	66 482	421	55.71	93.68	83.12
S3	80 299	75 525	67 674	66 047	410	55.60	93.60	82.25
S4	60 696	56 920	50 780	50 403	421	55.45	93.59	83.04
S6	80 110	74 968	67 346	64 208	414	54.79	93.59	80.15
R1	80 088	75 167	67 274	62 304	417	53.72	93.30	77.79
R2	80 074	75 096	67 657	64 078	414	54.75	93.50	80.02
R3	75 853	70 702	62 696	61 758	419	56.64	93.47	81.42
R4	80 141	75 989	68 176	61 433	423	53.18	93.72	76.66
R5	79 748	76 114	68 622	62 850	425	51.42	93.57	78.81
R6	79 574	75 541	67 609	65 096	420	55.69	93.78	81.81

样品名称 Sample ID	双端reads PE reads	原始序列 Raw tag	优化序列 Clean tag	有效序列 Effective tag	平均长度 Avglen/bp	GC/%	Q30/%	有效性 Effective/%
S1	80 094	75 613	67 304	65 710	420	56.36	93.52	82.04
S2	79 987	75 747	67 761	66 482	421	55.71	93.68	83.12
S3	80 299	75 525	67 674	66 047	410	55.60	93.60	82.25
S4	60 696	56 920	50 780	50 403	421	55.45	93.59	83.04
S6	80 110	74 968	67 346	64 208	414	54.79	93.59	80.15
R1	80 088	75 167	67 274	62 304	417	53.72	93.30	77.79
R2	80 074	75 096	67 657	64 078	414	54.75	93.50	80.02
R3	75 853	70 702	62 696	61 758	419	56.64	93.47	81.42
R4	80 141	75 989	68 176	61 433	423	53.18	93.72	76.66
R5	79 748	76 114	68 622	62 850	425	51.42	93.57	78.81
R6	79 574	75 541	67 609	65 096	420	55.69	93.78	81.81

样品编号 Sample ID	OTUs 数量 Number of OTUs	Ace 指数 Ace index	Chao 1 指数 Chao 1 index	Simpson 指数 Simpson index	Shannon 指数 Shannon index	覆盖度 Coverage
S1	1 199	1 245.397 0	1 264.578 0	0.006 0	6.121 7	0.997 6
S2	1 258	1 309.514 0	1 332.938 0	0.009 2	5.945 3	0.997 5
S3	987	1 208.833 0	1 211.467 0	0.530 6	2.094 9	0.994 8
S4	1 223	1 296.1970	1 321.057 0	0.009 1	5.844 7	0.995 7
S6	772	909.499 3	906.166 8	0.136 0	3.936 7	0.996 5
R1	889	1 109.902 0	1 113.591 0	0.148 8	3.587 3	0.994 5
R2	933	1 154.560 0	1 139.816 0	0.273 4	3.153 9	0.994 8
R3	1 146	1 229.390 0	1247.261 0	0.006 8	5.911 5	0.996 7
R4	784	942.978 4	959.035 3	0.130 4	3.940 7	0.995 8
R5	564	901.055 0	807.142 9	0.144 7	3.286 0	0.996 0
R6	1 041	1 127.204 0	1 160.436 0	0.009 9	5.721 2	0.996 5