Isolation, identification and pathogenicity determination of seed-borne fungi in Glycyrrhiza uralensis Fisch.

doi:10.3969/j.issn.1004-1524.20250207

Acta Agriculturae Zhejiangensis ›› 2026, Vol. 38 ›› Issue (3): 506-519.DOI: 10.3969/j.issn.1004-1524.20250207

• Plant Protection • Previous Articles Next Articles

Isolation, identification and pathogenicity determination of seed-borne fungi in Glycyrrhiza uralensis Fisch.

CHEN Runli¹(), LIU Pengfei¹, TANG Siyin², LOU Binggan¹^,^*()

^1. Ministry of Agriculture and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insect Pests, Institute of Biotechnology, Zhejiang University, Hangzhou 310058, China
^2. Xinjiang Huafang Chinese Herbal Medicine Co., Ltd., Yuli 841500, Xinjiang, China

Received:2025-03-20 Online:2026-03-25 Published:2026-04-17
Contact: LOU Binggan

Abstract

Abstract:

Glycyrrhiza uralensis Fisch., a valuable traditional Chinese herb primarily cultivated in northwest China, faces challenges including low seed germination, poor seedling emergence, and susceptibility to disease during the seedling stage, which hinder its industrial development. To assess the influence of seed-associated fungi on the germination and seedling establishment of licorice, fungal communities from G. uralensis Fisch. seeds collected across 19 sites in Xinjiang, Inner Mongolia, Shaanxi, and Gansu were isolated, characterized using morphological and molecular methods, the dominant fungi identified were Rhizopus arrhizus, Aspergillus flavus, Aspergillus niger, Alternaria tenuissima, Neopestalotiopsis clavispora, and Fusarium equiseti. Petri dish assays indicated that R. arrhizus, A. niger, and A. flavus significantly reduced seedling establishment at an inoculum level of 5 spores per seed, whereas N. clavispora and A. tenuissima required 300 spores per seed to produce a similar effect. The seedling emergence rate of licorice remained unaffected even when the spore load of F. equiseti reached 600 per seed. Pot-based pathogenicity tests further demonstrated that only R. arrhizus and A. flavus significantly affected the emergence and establishment rates of licorice seedlings. At a spore load of 50 spores per seed of R. arrhizus, both the emergence rate and seedling establishment rate of licorice decreased. When the spore load of A. flavus reached 300 spores per seed, significant impacts were observed on the emergence rate, seedling establishment rate, plant height, root length and biomass of licorice. This study provided essential technical support for the prevention and control of seedling diseases in licorice cultivation.

Key words: Glycyrrhiza uralensis Fisch., Rhizopus arrhizus, Aspergillus flavus, seedling disease, seed-borne fungi

CLC Number:

S435.672

CHEN Runli, LIU Pengfei, TANG Siyin, LOU Binggan. Isolation, identification and pathogenicity determination of seed-borne fungi in Glycyrrhiza uralensis Fisch.[J]. Acta Agriculturae Zhejiangensis, 2026, 38(3): 506-519.

Figures/Tables 11

References 44

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甘草种类Species of licorice	种子编号Seed No.	种子来源Seed source
乌拉尔甘草Glycyrrhiza uralensis Fisch.	W-XJ-B	新疆库尔勒市Korla, Xinjiang
	W-BZ-1	新疆博湖县Bohu, Xinjiang
	W-BZ-2	新疆轮台县Luntai, Xinjiang
	W-YL-1	新疆伊宁市Yining, Xinjiang
	W-CF-1	内蒙古赤峰市Chifeng, Inner Mongolia
	W-H-1	内蒙古巴彦淖尔市Bayannur, Inner Mongolia
	W-H-2	内蒙古杭锦旗Hanggin Banner, Inner Mongolia
	W-H-3	内蒙古阿拉善盟Alagxa, Inner Mongolia
	W-GS-1	甘肃张掖市Zhangye, Gansu
	W-GS-2	甘肃武威市Wuwei, Gansu
	W-SB-1	陕西榆林市Yulin, Shaanxi
胀果甘草Glycyrrhiza inflata Bat.	Z-AKS-1	新疆阿克苏沙雅县Xayar, Aksu, Xinjiang
	Z-AKS-2	新疆阿拉尔幸福镇Xingfu Town, Aral, Xinjiang
	Z-AKS-3	新疆阿拉尔科克库勒镇Kekekule Town, Aral, Xinjiang
	Z-AKS-4	新疆阿拉尔新开岭镇Xinkailing Town, Aral, Xinjiang
光果甘草Glycyrrhiza glabra L.	G-AKS-1	新疆阿拉尔新开岭镇Xinkailing Town, Aral, Xinjiang
	G-YL-1	新疆伊宁胡地亚于孜镇Hudiya Yuzi Town, Yining, Xinjiang
	G-YL-2	新疆伊宁拜什墩农场Baishidun Farm, Yining, Xinjiang
腺毛甘草Glycyrrhiza glandulifera Ledeb.	X-YL-1	新疆伊宁拜什墩农场Baishidun Farm, Yining, Xinjiang

甘草种类Species of licorice	种子编号Seed No.	种子来源Seed source
乌拉尔甘草Glycyrrhiza uralensis Fisch.	W-XJ-B	新疆库尔勒市Korla, Xinjiang
	W-BZ-1	新疆博湖县Bohu, Xinjiang
	W-BZ-2	新疆轮台县Luntai, Xinjiang
	W-YL-1	新疆伊宁市Yining, Xinjiang
	W-CF-1	内蒙古赤峰市Chifeng, Inner Mongolia
	W-H-1	内蒙古巴彦淖尔市Bayannur, Inner Mongolia
	W-H-2	内蒙古杭锦旗Hanggin Banner, Inner Mongolia
	W-H-3	内蒙古阿拉善盟Alagxa, Inner Mongolia
	W-GS-1	甘肃张掖市Zhangye, Gansu
	W-GS-2	甘肃武威市Wuwei, Gansu
	W-SB-1	陕西榆林市Yulin, Shaanxi
胀果甘草Glycyrrhiza inflata Bat.	Z-AKS-1	新疆阿克苏沙雅县Xayar, Aksu, Xinjiang
	Z-AKS-2	新疆阿拉尔幸福镇Xingfu Town, Aral, Xinjiang
	Z-AKS-3	新疆阿拉尔科克库勒镇Kekekule Town, Aral, Xinjiang
	Z-AKS-4	新疆阿拉尔新开岭镇Xinkailing Town, Aral, Xinjiang
光果甘草Glycyrrhiza glabra L.	G-AKS-1	新疆阿拉尔新开岭镇Xinkailing Town, Aral, Xinjiang
	G-YL-1	新疆伊宁胡地亚于孜镇Hudiya Yuzi Town, Yining, Xinjiang
	G-YL-2	新疆伊宁拜什墩农场Baishidun Farm, Yining, Xinjiang
腺毛甘草Glycyrrhiza glandulifera Ledeb.	X-YL-1	新疆伊宁拜什墩农场Baishidun Farm, Yining, Xinjiang

基因 Gene	引物名称 Primer name	引物序列(5'→3') Primer sequence (5'→3')	参考文献 Reference
rDNA-ITS	ITS1	TCCGTAGGTGAACCTGCGG	[24]
	ITS4	TCCTCCGCTTATTGATATGC
LSU	LR0R	ACCCGCTGAACTTAAGC
	LR5	TCCTGAGGGAAACTTCG
tef1*	EF1-728F	CATCGAGAAGTTCGAGAAGG	[25]
	EF1-986R	TACTTGAAGGAACCCTTACC
	raTEF-F	TACAAGGGATGGAACAA	[26]
	raTEF-R	AGCAAACTTACAGGCAA
	anTEF-F	CCTTCAAGTACGCYTGGGTTC	[27]
	anTEF-R	TTCTTGGAGTCACCGGCAA
	afTEF-F	GTGAATTCGAGGCTGGT	[28]
	afTEF-R	GAGGGGAAGACGGAGAG

基因 Gene	引物名称 Primer name	引物序列(5'→3') Primer sequence (5'→3')	参考文献 Reference
rDNA-ITS	ITS1	TCCGTAGGTGAACCTGCGG	[24]
	ITS4	TCCTCCGCTTATTGATATGC
LSU	LR0R	ACCCGCTGAACTTAAGC
	LR5	TCCTGAGGGAAACTTCG
tef1*	EF1-728F	CATCGAGAAGTTCGAGAAGG	[25]
	EF1-986R	TACTTGAAGGAACCCTTACC
	raTEF-F	TACAAGGGATGGAACAA	[26]
	raTEF-R	AGCAAACTTACAGGCAA
	anTEF-F	CCTTCAAGTACGCYTGGGTTC	[27]
	anTEF-R	TTCTTGGAGTCACCGGCAA
	afTEF-F	GTGAATTCGAGGCTGGT	[28]
	afTEF-R	GAGGGGAAGACGGAGAG

代表性菌株 Representative strain	基因名称 Gene name	序列长度/bp Sequence length/bp	GenBank登录号 GenBank accession number	比对结果 Alignment result	相似度/% Similarity/%
B-Z-3	ITS	604	PQ343918	黑曲霉	99.83
	tef1	457	PQ736917	Aspergillus niger	99.56
	LSU	960	PV682703		99.58
B-Z-4	ITS	554	PQ345466	新棒状拟盘多毛孢	99.46
	tef1	298	PV643062	Neopestalotiopsis clavispora	98.99
	LSU	938	PV682704		99.56
B-Z-6	ITS	599	PQ345471	黄曲霉	99.83
	tef1	384	PV643063	Aspergillus flavus	100.00
	LSU	949	PV682705		99.15
CF-Z-8	ITS	576	PQ345473	细极链格孢	99.83
	tef1	281	PV643064	Alternaria tenuissima	100.00
	LSU	928	PV682706		99.89
CF-Z-11	ITS	549	PQ345475	木贼镰刀菌	99.82
	tef1	285	PV643065	Fusarium equiseti	98.59
	LSU	916	PV682707		99.78
CF-Z-12	ITS	631	PQ345479	少根根霉	99.84
	tef1	475	PV643066	Rhizopus arrhizus	99.37
	LSU	1 035	PV682708		98.07

Isolation, identification and pathogenicity determination of seed-borne fungi in Glycyrrhiza uralensis Fisch.

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甘草种类 Species of licorice	种子编号 Seed umber	种子来源 Seed source	每粒种子孢子负荷量 Spore load per seed	菌株数(占比/%) Strain count(Percentage/%)
甘草种类 Species of licorice	种子编号 Seed umber	种子来源 Seed source	每粒种子孢子负荷量 Spore load per seed	黄曲霉 Aspergillus flavus	黑曲霉 Aspergillus niger	少根根霉 Rhizopus arrhizus
乌拉尔甘草 Glycyrrhiza uralensis Fisch.	W-XJ-B	新疆库尔勒市Korla, Xinjiang	60.83	33(45.21)	40(54.79)	0(0)
	W-BZ-1	新疆博湖县Bohu, Xinjiang	2.08	2(40.00)	2(40.00)	1(20.00)
	W-BZ-2	新疆轮台县Luntai, Xinjiang	2.92	3(42.86)	2(28.57)	2(28.57)
	W-YL-1	新疆伊宁市Yining, Xinjiang	5.00	4(33.33)	3(25.00)	5(41.67)
	W-CF-1	内蒙古赤峰市Chifeng, Inner Mongolia	2.92	2(28.57)	3(42.86)	2(28.57)
	W-H-1	内蒙古巴彦淖尔市Bayannur, Inner Mongolia	3.33	2(25.00)	5(62.50)	1(12.50)
	W-H-2	内蒙古杭锦旗Hanggin Banner, Inner Mongolia	4.17	4(40.00)	6(60.00)	0(0)
	W-H-3	内蒙古阿拉善盟Alagxa, Inner Mongolia	8.75	6(28.57)	10(47.62)	5(23.81)
	W-GS-1	甘肃张掖市Zhangye, Gansu	4.17	4(40.00)	5(50.00)	1(10.00)
	W-GS-2	甘肃武威市Wuwei, Gansu	10.00	8(33.33)	7(29.17)	9(37.50)
	W-SB-1	陕西榆林市Yulin, Shaanxi	5.83	4(28.57)	5(35.71)	5(35.71)
胀果甘草 Glycyrrhiza inflata Bat.	Z-AKS-1	新疆阿克苏沙雅县Xayar, Aksu, Xinjiang	6.67	6(37.50)	4(25.00)	6(37.50)
	Z-AKS-2	新疆阿拉尔幸福镇Xingfu Town, Aral, Xinjiang	9.17	7(31.82)	11(50.00)	4(18.18)
	Z-AKS-3	新疆阿拉尔科克库勒镇Kekekule Town, Aral, Xinjiang	8.33	4(20.00)	9(45.00)	7(35.00)
	Z-AKS-4	新疆阿拉尔新开岭镇Xinkailing Town, Aral, Xinjiang	5.42	5(38.46)	6(46.15)	2(15.38)
光果甘草 Glycyrrhiza glabra L.	G-AKS-1	新疆阿拉尔新开岭镇Xinkailing Town, Aral, Xinjiang	6.25	3(20.00)	8(53.33)	4(26.67)
	G-YL-1	新疆伊宁胡地亚于孜镇Hudiya Yuzi Town, Yining, Xinjiang	8.75	5(23.81)	12(57.14)	4(19.05)
	G-YL-2	新疆伊宁拜什墩农场Baishidun Farm, Yining, Xinjiang	10.00	5(20.83)	14(58.33)	5(20.83)
腺毛甘草 Glycyrrhiza glandulifera Ledeb.	X-YL-1	新疆伊宁拜什墩农场Baishidun Farm, Yining, Xinjiang	2.92	2(28.57)	4(57.14)	1(14.29)

甘草种类 Species of licorice	种子编号 Seed number	种子来源 Seed source	带菌率/% Infection rate/%	菌株数(占比/%) Strain count(Percentage/%)
甘草种类 Species of licorice	种子编号 Seed number	种子来源 Seed source	带菌率/% Infection rate/%	黄曲霉 Aspergillus flavus	黑曲霉 Aspergillus niger	少根根霉 Rhizopus arrhizus	细极链格孢 Alternaria tenuissima	木贼镰刀菌 Fusarium equiseti	新棒状拟盘多毛孢 Neopestalotiopsis clavispora
乌拉尔甘草 Glycyrrhiza uralensis Fisch.	W-XJ-B	新疆库尔勒市 Korla, Xinjiang	12.96	0(0)	4(28.57)	0(0)	8(57.14)	0(0)	2(14.29)
	W-BZ-1	新疆博湖县 Bohu, Xinjiang	18.52	2(10.00)	3(15.00)	4(20.00)	11(55.00)	0(0)	0(0)
	W-BZ-2	新疆轮台县 Luntai, Xinjiang	21.30	4(17.39)	2(8.70)	3(13.04)	13(56.52)	0(0)	1(4.35)
	W-YL-1	新疆伊宁市 Yining, Xinjiang	31.48	3(8.82)	5(14.71)	4(11.76)	20(58.82)	0(0)	2(5.88)
	W-CF-1	内蒙古赤峰市 Chifeng, Inner Mongolia	37.96	2(4.88)	13(31.71)	0(0)	24(58.54)	2(4.88)	0(0)
	W-H-1	内蒙古巴彦淖尔市 Bayannur, Inner Mongolia	29.63	4(12.50)	6(18.75)	5(15.63)	14(43.75)	3(9.38)	0(0)
	W-H-2	内蒙古杭锦旗 Hanggin Banner, Inner Mongolia	26.85	3(10.34)	3(10.34)	4(13.79)	18(62.07)	1(3.45)	0(0)
	W-H-3	内蒙古阿拉善盟 Alagxa, Inner Mongolia	25.93	4(14.29)	5(17.86)	3(10.71)	16(57.14)	0(0)	0(0)
	W-GS-1	甘肃张掖市 Zhangye, Gansu	37.96	6(14.63)	8(19.51)	4(9.76)	21(51.22)	0(0)	2(4.88)
	W-GS-2	甘肃武威市 Wuwei, Gansu	21.30	3(13.04)	4(17.39)	2(8.70)	13(56.52)	0(0)	1(4.35)
	W-SB-1	陕西榆林市 Yulin, Shaanxi	15.74	2(11.76)	3(17.65)	2(11.67)	6(35.29)	2(11.67)	2(11.67)
胀果甘草 Glycyrrhiza inflata Bat.	Z-AKS-1	新疆阿克苏沙雅县 Xayar, Aksu, Xinjiang	12.04	3(23.08)	2(15.38)	1(7.69)	7(53.85)	0(0)	0(0)
	Z-AKS-2	新疆阿拉尔幸福镇 Xingfu Town, Aral, Xinjiang	20.37	2(9.09)	5(22.73)	4(18.18)	11(50.00)	0(0)	0(0)
	Z-AKS-3	新疆阿拉尔科克库勒镇 Kekekule Town, Aral, Xinjiang	19.44	3(14.29)	4(19.05)	4(19.05)	9(42.86)	1(4.76)	0(0)
	Z-AKS-4	新疆阿拉尔新开岭镇 Xinkailing Town, Aral, Xinjiang	23.15	6(24.00)	2(8.00)	3(12.00)	12(48.00)	2(8.00)	0(0)
光果甘草 Glycyrrhiza glabra L.	G-AKS-1	新疆阿拉尔新开岭镇 Xinkailing Town, Aral, Xinjiang	13.89	3(20.00)	6(40.00)	3(20.00)	3(20.00)	0(0)	0(0)
	G-YL-1	新疆伊宁胡地亚于孜镇 Hudiya Yuzi Town, Yining, Xinjiang	16.67	5(27.78)	4(22.22)	3(16.67)	6(33.33)	0(0)	0(0)
	G-YL-2	新疆伊宁拜什墩农场 Baishidun Farm,Yining, Xinjiang	25.00	8(29.63)	3(11.11)	5(18.52)	11(40.74)	0(0)	0(0)
腺毛甘草 Glycyrrhiza glandulifera Ledeb.	X-YL-1	新疆伊宁拜什墩农场 Baishidun Farm, Yining, Xinjiang	10.19	3(27.27)	1(9.09)	2(18.18)	5(45.45)	0(0)	0(0)