Isolation and screening of cultivable endophytes in sifferent organs of Glycyrrhiza uralensis Fisch and preliminary screening of β-glucosidase producing strains

doi:10.6048/j.issn.1001-4330.2024.01.022

Xinjiang Agricultural Sciences ›› 2024, Vol. 61 ›› Issue (1): 199-208.DOI: 10.6048/j.issn.1001-4330.2024.01.022

• Agricultural Product Analysis and Detection·Forestry·Agricultural Information·Plant Protection • Previous Articles Next Articles

Isolation and screening of cultivable endophytes in sifferent organs of Glycyrrhiza uralensis Fisch and preliminary screening of β-glucosidase producing strains

KONG Xiaoshuang¹(), WEI Ran², DONG Yinghong³, HOU Min³(), Maierhaba Aihemaiti³, HOU Xinqiang³, YANG Wenqi³, CUI Weidong³()

1. College of Life Science and Technology,Xinjiang University,Urumqi 830052,China
2. College of Agriculture,Xinjiang Agricultural University,Urumqi 830052,China
3. Institute of Applied Microbiology,Xinjiang Academy of Agricultural Sciences,Xinjiang Special Environmental Microbiology Laboratory,Urumqi 830091,China

Received:2023-04-11 Online:2024-01-20 Published:2024-02-21
Correspondence author: HOU Min(1983-),Urumqi,female,master,researcher, research direction in development and application of feed microorganism,(E-mail) hmde_092@163.com; CUI Weidong(1969-),Urumqi,male,master,researcher,research direction in development and application of feed microorganism,(E-mail) cuwedo@163.com
Supported by:
Key R&D Task Projects in Xinjiang Uygur Autonomous Region(2022B02042);Key R&D Program Project of Xinjiang Uygur Autonomous Region(2022B02056-2)

乌拉尔甘草不同组织可培养内生菌分离筛选及产β-葡萄糖苷酶菌株初筛

孔晓双¹(), 魏然², 董应宏³, 侯敏³(), 买尔哈巴·艾合买提³, 侯新强³, 杨文琦³, 崔卫东³()

1.新疆大学生命科学与技术学院,乌鲁木齐 830052
2.新疆农业大学农学院,乌鲁木齐 830052
3.新疆农业科学院微生物应用研究所/新疆特殊环境微生物实验室,乌鲁木齐 830091

通讯作者: 侯敏(1983-),女,新疆乌鲁木齐人,研究员,硕士,研究方向为饲用微生物开发与应用,(E-mail)hmde_092@163.com; 崔卫东(1969-),男,新疆乌鲁木齐人,研究员,硕士,研究方向为饲用微生物开发与应用,(E-mail)cuwedo@163.com
作者简介:孔晓双(1998-),女,河南周口人,硕士研究生,研究方向为食品工程,(E-mail)1940693886@qq.com
基金资助:
新疆维吾尔自治区重点研发任务专项项目(2022B02042);新疆维吾尔自治区重点研发计划项目(2022B02056-2)

Abstract

Abstract:

【Objective】 To study the resource status of culturable endophytes from different parts of Glycyrrhiza uralensis Fisch Altay region of Xinjiang and to screen high-yield β-glucosidase strains. 【Methods】 Using Glycyrrhiza uralensis Fisch as the material,the endophytic bacteria in its taproot,fibrous root,stem and leaf were isolated,purified and identified by means of tissue homogenization combined with 16S rDNA and ITS-rDNA molecular biology methods. The β-glucosidase producing strains were screened by aescin medium and β-glucosidase activity determination. 【Results】 The results showed that 48 endophytes,33 endophytic bacteria and 15 endophytic fungi were isolated from Glycyrrhiza uralensis Fisch. The number of endophytic bacteria isolated from taproots and stems was the largest,and that of endophytic fungi colonized in taproots was the largest. The 33 endophytic bacteria from Glycyrrhiza uralensis Fisch belonged to 8 genera,and the most Bacillus were isolated,and the endophytic bacteria in the taproot of Glycyrrhiza uralensis Fisch were the most widely isolated species. The diversity in different parts was as follows:taproot > fibrous root = stem = leaf. 15 endophytic fungi from Glycyrrhiza uralensis Fisch belonged to 7 genera,Cladosporium was the dominant genus of endophytic fungi. The diversity of endophytic fungi in different parts was as follows:taproot > stem > leaf > fibrous root. A total of 12 strains had the ability to produce β-glucosidase,among which the most strains isolated from taproots,accounted for 41.7%. 【Conclusion】 The diversity of taproot organ of cultivable endophytes from Glycyrrhiza uralensis Fisch is rich and there are more β-glucosidase producing strains,which are the dominant part for breeding enzyme-producing strains.

Key words: Glycyrrhiza uralensis Fisch; cultivable endophytes; screening; β-glucosaccharase

摘要：

【目的】研究新疆阿勒泰地区乌拉尔甘草不同组织可培养内生菌的资源状况以及选育高产β-葡萄糖苷酶菌株。【方法】以乌拉尔甘草为材料,运用组织匀浆法与16S rDNA 和 ITS-rDNA分子生物学方法相结合方式,对其主根、须根、茎、叶中的内生菌进行分离、纯化和鉴定;采用七叶苷培养基和β-葡萄糖苷酶酶活力测定筛选产β-葡萄糖苷酶菌株。【结果】从甘草中分离获得内生菌共48株,其中内生细菌33株,内生真菌15株。在主根和茎部位分离内生细菌数量最多,内生真菌在主根中定殖的数量最多。33株甘草内生细菌隶属于8个属,芽孢杆菌属菌株分离获得最多,且甘草主根部位的内生细菌分离种类最多,在甘草不同组织中多样性表现为主根>须根=茎=叶。15株甘草内生真菌隶属于7个属,枝孢属是甘草内生真菌的优势属,内生真菌在不同组织中多样性表现为主根>茎>叶>须根。共有12株具有产β-葡萄糖苷酶的能力,其中分离自主根部位的菌株最多,占比41.7%。【结论】甘草主根组织的可培养内生菌的多样性相较其他组织丰富,且主根部位产β-葡萄糖苷酶菌株较多,是选育产酶菌株的优势部位。

关键词: 乌拉尔甘草, 可培养内生菌, 筛选, β-葡萄糖苷酶

CLC Number:

S188
S812

KONG Xiaoshuang, WEI Ran, DONG Yinghong, HOU Min, Maierhaba Aihemaiti, HOU Xinqiang, YANG Wenqi, CUI Weidong. Isolation and screening of cultivable endophytes in sifferent organs of Glycyrrhiza uralensis Fisch and preliminary screening of β-glucosidase producing strains[J]. Xinjiang Agricultural Sciences, 2024, 61(1): 199-208.

孔晓双, 魏然, 董应宏, 侯敏, 买尔哈巴·艾合买提, 侯新强, 杨文琦, 崔卫东. 乌拉尔甘草不同组织可培养内生菌分离筛选及产β-葡萄糖苷酶菌株初筛[J]. 新疆农业科学, 2024, 61(1): 199-208.

Figures/Tables 7

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部位 Part	内生细菌 Endophytic bacteria		内生真菌 Endophytic fungi
部位 Part	菌株数 Strain No.(个)	百分率 Percentage(%)	菌株数 Strain No.(个)	百分率 Percentage(%)
主根 Taproot	9	27.3	8	53.3
须根 Fibrous root	8	24.2	1	6.7
茎 Stem	9	27.3	4	26.7
叶 Leaf	7	21.2	2	13.3

部位 Part	内生细菌 Endophytic bacteria		内生真菌 Endophytic fungi
部位 Part	菌株数 Strain No.(个)	百分率 Percentage(%)	菌株数 Strain No.(个)	百分率 Percentage(%)
主根 Taproot	9	27.3	8	53.3
须根 Fibrous root	8	24.2	1	6.7
茎 Stem	9	27.3	4	26.7
叶 Leaf	7	21.2	2	13.3

菌株 Strains	部位 Part	同源性最高的菌株 Closest species	同源性 Samilarity(%)	GenBank登录号 GenBank accession No.
A12	主根	Bacillus velezensis CR-502	99.43	OP435752
A14	茎	Fictibacillus nanhaiensis JSM 082006	99.16	OP435753
A15	茎	Bacillus amyloliquefaciens DSM 7	99.79	OP435754
A17	叶	Bacillus rugosus SPB7	99.52	ON366397
A22	叶	Bacillus velezensis CR-502	99.71	OP435755
A23	须根	Bacillus rugosus SPB7	97.38	OP435756
A28	主根	Bacillus velezensis CR-502	99.50	OP435757
A29	主根	Bacillus velezensis CR-502	99.43	OP435758
A30	主根	Bacillus subtilis 3610	99.20	OP435759
A31	茎	Bacillus cabrialesii TE3	99.11	OP435760
A33	茎	Bacillus tequilensis KCTC 13622	97.06	OP435761
A34	茎	Bacillus velezensis CR-502	99.29	OP435762
A36	叶	Janthinobacterium svalbardensis JA-1	99.36	OP435763
A37	叶	Bacillus tequilensis KCTC 13622	99.59	OP435764
A41	须根	Pseudomonas piscicoa dhj-6	99.92	OP435765
A44	须根	Bacillus tequilensis KCTC 13622	99.45	OP435766
A45	须根	Agrobacterium radiobacter ATCC19358	99.04	OP435767
A62	主根	Fictibacillus nanhaiensis JSM 082006	95.37	OP435768
A63	主根	Acinetobacter oryzae B23	98.95	OP435769
A65	主根	Bacillus subtilis 3610	99.14	ON366398
A71	茎	Fictibacillus nanhaiensisJSM 082006	99.16	OP435770
A72	茎	Bacillus cabrialesii TE3	99.31	OP435771
A73	茎	Metabacillus litoralis SW-211	94.55	OP435772
A75	叶	Niallia circulans ATCC 4513	99.24	OP435773
A78	须根	Bacillus rugosus SPB7	99.38	ON366399
A79	主根	Janthinobacterium svalbardensis JA-1	94.96	OP435774
A80	主根	Bacillus tequilensis KCTC 13622	99.66	OP435775
A81	茎	Bacillus tequilensis KCTC 13622	96.96	OP435776
A82	叶	Bacillus cabrialesii TE3	99.31	OP435777
A83	叶	Bacillus rugosus SPB7	99.31	OP435778
A84	须根	Bacillus mobilis 0711P9-1	99.52	OP435779
A85	须根	Bacillus tequilensis KCTC 13622	99.18	OP435780
A86	须根	Bacillus rugosus SPB7	96.83	OP435781

菌株 Strains	部位 Part	同源性最高的菌株 Closest species	同源性 Samilarity(%)	GenBank登录号 GenBank accession No.
A12	主根	Bacillus velezensis CR-502	99.43	OP435752
A14	茎	Fictibacillus nanhaiensis JSM 082006	99.16	OP435753
A15	茎	Bacillus amyloliquefaciens DSM 7	99.79	OP435754
A17	叶	Bacillus rugosus SPB7	99.52	ON366397
A22	叶	Bacillus velezensis CR-502	99.71	OP435755
A23	须根	Bacillus rugosus SPB7	97.38	OP435756
A28	主根	Bacillus velezensis CR-502	99.50	OP435757
A29	主根	Bacillus velezensis CR-502	99.43	OP435758
A30	主根	Bacillus subtilis 3610	99.20	OP435759
A31	茎	Bacillus cabrialesii TE3	99.11	OP435760
A33	茎	Bacillus tequilensis KCTC 13622	97.06	OP435761
A34	茎	Bacillus velezensis CR-502	99.29	OP435762
A36	叶	Janthinobacterium svalbardensis JA-1	99.36	OP435763
A37	叶	Bacillus tequilensis KCTC 13622	99.59	OP435764
A41	须根	Pseudomonas piscicoa dhj-6	99.92	OP435765
A44	须根	Bacillus tequilensis KCTC 13622	99.45	OP435766
A45	须根	Agrobacterium radiobacter ATCC19358	99.04	OP435767
A62	主根	Fictibacillus nanhaiensis JSM 082006	95.37	OP435768
A63	主根	Acinetobacter oryzae B23	98.95	OP435769
A65	主根	Bacillus subtilis 3610	99.14	ON366398
A71	茎	Fictibacillus nanhaiensisJSM 082006	99.16	OP435770
A72	茎	Bacillus cabrialesii TE3	99.31	OP435771
A73	茎	Metabacillus litoralis SW-211	94.55	OP435772
A75	叶	Niallia circulans ATCC 4513	99.24	OP435773
A78	须根	Bacillus rugosus SPB7	99.38	ON366399
A79	主根	Janthinobacterium svalbardensis JA-1	94.96	OP435774
A80	主根	Bacillus tequilensis KCTC 13622	99.66	OP435775
A81	茎	Bacillus tequilensis KCTC 13622	96.96	OP435776
A82	叶	Bacillus cabrialesii TE3	99.31	OP435777
A83	叶	Bacillus rugosus SPB7	99.31	OP435778
A84	须根	Bacillus mobilis 0711P9-1	99.52	OP435779
A85	须根	Bacillus tequilensis KCTC 13622	99.18	OP435780
A86	须根	Bacillus rugosus SPB7	96.83	OP435781

菌株 Strains	部位 Part	同源性最高的菌株 Closest species	同源性 Samilarity(%)	GenBank登录号 GenBank accession No.
p₂	主根	Penicillium sp. ST-PSB-J1	99.83	OP681416
p₃	主根	Aspergillus fumigatus C1946	99.83	OP681417
p₄	主根	Cladosporium cladosporioides 08SK030	92.03	OP681418
p₆	主根	Alternaria alternata JN10	99.81	OP681419
p₉	茎	Candolleomyces candolleanus NW550	99.42	ON529501
p₁₀	茎	Penicillium cyclopium CICC 4026	99.15	OP681420
p₁₁	茎	Cladosporium fusiforme a2	100.00	OP681421
p₁₂	茎	Aspergillus fumigatus MUST	99.15	OP681422
p₁₃	主根	Cladosporium cladosporioides 08SK030	99.51	OP681423
p₁₅	须根	Fusarium oxysporum NDJ2	99.65	OP681424
p₁₆	主根	Aspergillus fumigatus MUST	100.00	OP681425
p₁₇	叶	Cladosporium perangustum IA39	97.42	OP681426
p₃₆	主根	Trichothecium sp. 10731	99.42	OP681427
p₃₈	主根	Alternaria alstroemeriae	96.85	OP681428
p₄₄	叶	Penicillium glabrum PM2	99.64	OP681429

Isolation and screening of cultivable endophytes in sifferent organs of Glycyrrhiza uralensis Fisch and preliminary screening of β-glucosidase producing strains

乌拉尔甘草不同组织可培养内生菌分离筛选及产β-葡萄糖苷酶菌株初筛

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