楸树组培苗内生菌分离鉴定与功能特性解析

doi:10.11924/j.issn.1000-6850.casb2024-0093

中国农学通报 ›› 2024, Vol. 40 ›› Issue (31): 7-13.doi: 10.11924/j.issn.1000-6850.casb2024-0093

楸树组培苗内生菌分离鉴定与功能特性解析

王璐瑶¹(), 郭泽亮¹, 陈卫卫¹, 穆艳娟², 解孝满², 仝伯强², 程仕伟¹(), 鲁仪增²()

¹ 鲁东大学生命科学学院，山东烟台 264025
² 山东省林草种质资源中心，济南 250102

收稿日期:2024-02-06 修回日期:2024-08-02 出版日期:2024-11-05 发布日期:2024-11-04
通讯作者:
程仕伟，男，1976年出生，山东潍坊人，副教授，博士，研究方向为微生物资源利用。通信地址：264025 山东烟台芝罘区红旗中路186号鲁东大学生命科学学院，Tel：0535-6685003，E-mail：swbiotech@yeah.net。
鲁仪增，男，1978年出生，山东菏泽人，正高级工程师，博士，研究方向为草种质资源保护与创新应用。通信地址：250102 山东济南历城区港九路2011号山东省林草种质资源中心，Tel：0531-88557843，E-mail：luyizeng@126.com。
作者简介:
王璐瑶，女，1999年出生，山东潍坊人，硕士研究生，研究方向为植物内生菌互作。通信地址：264025 山东烟台芝罘区红旗中路186号鲁东大学生命科学学院，Tel：0535-6685003，E-mail：wangly0102@163.com。
基金资助:
山东省重点研发计划（重大科技创新工程）项目“珍贵用材树种种质资源挖掘与精准鉴定”(2021LZGC02304)

Isolation and Identification of Endophytic Bacteria in Catalpa bungei Tissue Culture Seedling and Its Functional Characteristic Analysis

WANG Luyao¹(), GUO Zeliang¹, CHEN Weiwei¹, MU Yanjuan², XIE Xiaoman², TONG Boqiang², CHENG Shiwei¹(), LU Yizeng²()

¹ School of Life Sciences, Ludong University, Yantai, Shandong 264025
² Shandong Provincial Center of Forest and Grass Germplasm Resources, Jinan 250102

Received:2024-02-06 Revised:2024-08-02 Published:2024-11-05 Online:2024-11-04

摘要/Abstract

摘要：

研究旨在挖掘楸树组培苗内生菌资源，开发具有应用潜力的农业微生物菌群，从而促进组培苗快速生长和繁殖。利用选择性培养基对楸树组培苗内生菌进行分离鉴定。采用Ashby固体培养基初筛，测定液体培养基中有效氮浓度，评价内生菌固氮效果；通过Nbrip固体培养基初筛，测定液体培养基中可溶性磷浓度，评价内生菌解磷效果；测定内生菌对植物病原菌链格孢菌和葱柄锈菌的抑制效果。自楸树组培苗中分离获得24株内生菌，经形态和16S rRNA序列鉴定均为根瘤菌属的Rhizobium zeae。菌株NF6的固氮能力最强，其培养基中有效氮浓度达111.97 mg/L；菌株NF11的解磷效果最好，其培养基中可溶性磷浓度达1007.95 mg/L；多数内生菌对供试病原菌均有抑制效果，分别有6株和2株内生菌对链格孢菌和葱柄锈菌的抑制率超过20%。本研究获得了植物促生和生防效果的菌株资源，可为组培苗内生菌的潜在应用提供理论依据。

关键词: 楸树, 组培苗, 内生菌, 分离鉴定, 功能特性, 固氮作用, 解磷作用

Abstract:

This study aimed to explore the endophytic bacterial resources of Catalpa bungei tissue culture seedlings, to develop agricultural microorganisms with application potential, promoting the rapid growth and propagation of tissue culture seedlings. The endophytic bacteria were isolated and identified from C. bungei tissue culture seedlings by selective medium in this paper. The nitrogen content in the culture solution was determined to evaluate the nitrogen fixing effect after preliminary screening with Ashby solid medium, and the phosphorus solubilization effect was evaluated using phosphorus content in the culture solution after preliminary screening with Nbrip solid medium, then the inhibitory effect of endophytic bacteria was measured with Alternaria alternate and Fomitopsis semiolaccatus as test objects. 24 strains of endophytic bacteria were isolated from the seedling of C. bungei, and they were identified as Rhizobium zeae by morphological and 16S rRNA sequence analysis. The results indicated that the nitrogen fixing capacity of strain NF6 was best as its nitrogen concentration in the Ashby liquid medium reached 111.97 mg/L, and strain NF11 had the best phosphorus solubilization effect due to the phosphorus concentration in Nbrip culture medium reached 1007.95 mg/L. Furthermore, most of the endophytic R. zeae had inhibitory effects on the tested pathogens, and more than 20% of inhibitory rates against A. alternate and F. semiolaccatus respectively had obtained for the 6 and 2 strains of endophytic R. zeae. In this study, the resources of endophytic bacteria with the potentials of promoting growth and resistance to pathogens were obtained, which provided theoretical basis for the potential application of endophytic bacteria in tissue culture seedlings.

Key words: Catalpa bungei, tissue culture seedling, endophyte, isolation and identification, functional characteristics, nitrogen fixation, phosphorus solubilization

王璐瑶, 郭泽亮, 陈卫卫, 穆艳娟, 解孝满, 仝伯强, 程仕伟, 鲁仪增. 楸树组培苗内生菌分离鉴定与功能特性解析[J]. 中国农学通报, 2024, 40(31): 7-13.

WANG Luyao, GUO Zeliang, CHEN Weiwei, MU Yanjuan, XIE Xiaoman, TONG Boqiang, CHENG Shiwei, LU Yizeng. Isolation and Identification of Endophytic Bacteria in Catalpa bungei Tissue Culture Seedling and Its Functional Characteristic Analysis[J]. Chinese Agricultural Science Bulletin, 2024, 40(31): 7-13.

图/表 7

参考文献 30

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菌株序号	菌落直径/mm	菌株序号	菌落直径/mm
NF1	5.45±0.02	NF13	4.05±0.25
NF2	6.00±0.08	NF14	5.07±0.07
NF3	5.38±0.08	NF15	3.88±0.24
NF4	5.63±0.01	NF16	4.05±0.03
NF5	5.37±0.01	NF17	5.25±0.03
NF6	7.00±0.07	NF18	5.39±0.07
NF7	5.75±0.15	NF19	4.88±0.05
NF8	5.13±0.10	NF20	6.32±0.19
NF9	3.38±0.05	NF21	7.63±0.15
NF10	4.87±0.05	NF22	6.87±0.28
NF11	3.75±0.12	NF23	6.13±0.35
NF12	3.38±0.11	NF24	7.50±0.09

菌株序号	菌落直径/mm	菌株序号	菌落直径/mm
NF1	5.45±0.02	NF13	4.05±0.25
NF2	6.00±0.08	NF14	5.07±0.07
NF3	5.38±0.08	NF15	3.88±0.24
NF4	5.63±0.01	NF16	4.05±0.03
NF5	5.37±0.01	NF17	5.25±0.03
NF6	7.00±0.07	NF18	5.39±0.07
NF7	5.75±0.15	NF19	4.88±0.05
NF8	5.13±0.10	NF20	6.32±0.19
NF9	3.38±0.05	NF21	7.63±0.15
NF10	4.87±0.05	NF22	6.87±0.28
NF11	3.75±0.12	NF23	6.13±0.35
NF12	3.38±0.11	NF24	7.50±0.09

菌株序号	解磷圈与菌落直径比	菌株序号	解磷圈与菌落直径比
NF1	—	NF13	2.25±0.12
NF2	2.42±0.04	NF14	2.81±0.53
NF3	2.21±0.58	NF15	0.82±0.16
NF4	2.25±0.23	NF16	1.74±0.10
NF5	1.77±0.06	NF17	1.17±0.02
NF6	2.02±0.33	NF18	2.28±0.48
NF7	1.80±0.52	NF19	1.94±0.08
NF8	1.28±0.12	NF20	2.31±0.81
NF9	1.72±0.43	NF21	2.52±0.88
NF10	1.72±0.12	NF22	2.70±0.17
NF11	1.89±0.41	NF23	2.20±0.37
NF12	2.23±0.65	NF24	1.82±0.01

菌株序号	解磷圈与菌落直径比	菌株序号	解磷圈与菌落直径比
NF1	—	NF13	2.25±0.12
NF2	2.42±0.04	NF14	2.81±0.53
NF3	2.21±0.58	NF15	0.82±0.16
NF4	2.25±0.23	NF16	1.74±0.10
NF5	1.77±0.06	NF17	1.17±0.02
NF6	2.02±0.33	NF18	2.28±0.48
NF7	1.80±0.52	NF19	1.94±0.08
NF8	1.28±0.12	NF20	2.31±0.81
NF9	1.72±0.43	NF21	2.52±0.88
NF10	1.72±0.12	NF22	2.70±0.17
NF11	1.89±0.41	NF23	2.20±0.37
NF12	2.23±0.65	NF24	1.82±0.01

菌株序号	链格孢菌	葱柄锈菌
NF1	11.40±0.72	8.89±0.39
NF2	19.30±1.21	11.11±0.24
NF3	13.16±0.53	7.78±0.31
NF4	21.05±1.48	11.11±0.67
NF5	22.81±0.82	2.22±0.12
NF6	19.30±1.66	24.44±2.15
NF7	9.65±0.19	6.67±0.41
NF8	12.28±0.28	11.11±0.81
NF9	23.68±1.49	12.22±0.56
NF10	18.42±1.94	20.00±2.38
NF11	11.40±049	8.89±0.60
NF12	21.05±2.28	6.67±0.29
NF13	21.05±1.55	18.89±1.09
NF14	18.42±6.17	15.56±1.44
NF15	2.63±0.69	4.44±0.67
NF16	17.54±2.57	10.00±2.55
NF17	14.04±1.41	17.78±3.09
NF18	10.53±1.77	8.89±1.23
NF19	12.28±0.89	—
NF20	8.77±1.46	—
NF21	8.77±2.05	3.33±0.74
NF22	21.05±3.19	15.56±2.69
NF23	14.04±1.82	11.11±2.71
NF24	—	—

楸树组培苗内生菌分离鉴定与功能特性解析

Isolation and Identification of Endophytic Bacteria in Catalpa bungei Tissue Culture Seedling and Its Functional Characteristic Analysis

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