地黄不同时期内生促生细菌的筛选及其促生特性分析

doi:10.13560/j.cnki.biotech.bull.1985.2021-0964

摘要/Abstract

摘要：

从地黄中开发优势菌种资源并分析促生特性,为开发地黄微生物专用促生菌肥提供基础。采用稀释平板法,以地黄不同时期根、茎、叶为材料分离内生细菌,根据形态学、生理生化、16S rRNA分析进行分类归属。选出54株内生细菌对产IAA、铁载体活性,溶磷以及固氮能力进行了评价,结果显示在不同时期共分离出512株内生细菌,将其归类为54组,每组选出1株进行检测,其中41株具有产IAA的能力;29株具有溶磷能力;28株具有产铁载体能力;25株具有固氮能力。该结果为研究地黄专用微生物促生菌肥提供参考,并为后续微生物促生菌肥应用到地黄大田实验中奠定基础。

关键词: 地黄, 分离鉴定, 内生细菌, 促生

Abstract:

The development of dominant strain resources from Rehmannia glutinosa and the analysis of growth promoting characteristics will provide a basis for the development of special growth-promoting bacterial fertilizer for R. glutinosa microorganisms. Dilution plate method was used to classify the endophytic bacteria isolated from the roots,stems and leaves of R. glutinosa at different stages to their morphologies,physiologies and biochemistries and 16S rRNA analysis. And the selected 54 strains of endophytic bacteria were evaluated based on their IAA-yielding capacities,iron-carrier activities,P-dissolving capacities,and N-fixing capacities. The results showed that 512 strains of endophytic bacteria were isolated in different periods and classified into 54 groups. One strain in each group was selected for detection,of which 41 strains had the ability to produce IAA;29 strains of them had the ability of dissolving phosphorus;28 strains had the ability to produce iron carrier;and 25 strains had nitrogen fixation ability. This result provides a reference for the study of special microbial growth-promoting bacteria fertilizer for R. glutinosa,and lays a foundation for the subsequent application of microbial growth-promoting bacteria fertilizer to the field experiment of R. glutinosa.

Key words: Rehmannia glutinosa., isolation and identification, endophytic bacteria, growth-promoting

王春艳, 腊贵晓, 苏秀红, 李萌, 董诚明. 地黄不同时期内生促生细菌的筛选及其促生特性分析[J]. 生物技术通报, 2022, 38(4): 242-252.

WANG Chun-yan, LA Gui-xiao, SU Xiu-hong, LI Meng, DONG Cheng-ming. Screening of Endophytic Bacteria from Rehmannia glutinosa at Different Stages and Analysis of Their Growth-promoting Characteristics[J]. Biotechnology Bulletin, 2022, 38(4): 242-252.

图/表 8

参考文献 44

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菌株 Strain	最相近菌种 The closest strain	登录号 Login number	相似性 Similarity/%	菌株 Strain	最相近菌种 The closest strain	登录号 Login number	相似性 Similarity/%
ER1	Bacillus proteolyticus	NR157735	99.9	ER28	Bacillus paramycoides	MT124553	99.72
ER2	Bacillus methylotrophicus	KC790266	99.9	ER29	Pseudomonas plecoglossicida	MT124554	100.0
ER3	Bacillus toyoneusis	NR121761	100.0	ER30	Pseudomonas monteilii	MT124555	98.0
ER4	Bacillus licheniformis	MH842141	99.9	ER31	Pseudomonas taiwanensis	JF911384	100.0
ER5	Bacillus cereus	FJ501984	100.0	ER32	Pseudomonas aeruginosa	NR117678	100.0
ER6	Bacillus wiedmannii	MK088275	100.0	ER33	Pseudomonas fluorescens	MN256396	99.0
ER7	Bacillus amyloliquefacieus	KU291367	100.0	ER34	Pseudomonas oryzihabitans	JQ660200	100.0
ER8	Bacillus velezensis	MG996871	100.0	ER35	Pseudomonas psychrotolerans	KY882047	99.9
ER9	Bacillus siamensis	MN166187	100.0	ER36	Pseudomonas stutzeri	FJ768951	99.9
ER10	Bacillus subtilis	MK860021	100.0	ER37	Proteus mirabilis	MK332376	100.0
ER11	Bacillus anthracis	CP031642	99.9	ER38	Micrococcus luteus	MH045862	100.0
ER12	Bacillus aryabhattai	MK859946	100.0	ER39	Stenotrophomonas maltophilia	MH000623	100.0
ER13	Bacillus vallismortis	NR024696	100.0	ER40	Lysinibacillus parvibpronicapiens	NR114213	99.7
ER14	Bacillus oryzaecorticis	KU877642	99.9	ER41	Brachybacterium paraconglomeratum	NR025502	100.0
ER15	Bacillus aquimaris	KF358255	99.8	ER42	Agrobacterium tumefaciens	MN094859	99.9
ER16	Bacillus marisflavi	KX495310	99.8	ER43	Lysinibacillus fusiformis	KP980567	99.9
ER17	Bacillus sonerensis	MK860002	100.0	ER44	Enterobacter cloacae	KM538690	99.9
ER18	Bacillus thuringiensis	KJ769222	99.5	ER45	Enterobacter bugandensis	CP039453	99.8
ER19	Bacillus safensis	MK859936	100.0	ER46	Enterobacter hormaechei	CP027142	100.0
ER20	Bacillus simplex	KF818647	99.9	ER47	Enterobacter ludwigii	NR042349	99.9
ER21	Bacillus firmus	KP165527	99.9	ER48	Enterobacter xiangfangensis	MH200641	99.5
ER22	Bacillus altitudinis	KU663665	100.0	ER49	Enterobacter soli	NR117547	99.3
ER23	Bacillus flexus	KR999917	100.0	ER50	Terribacillus goriensis	CP008876	100.0
ER24	Bacillus horikoshii	CP020880	99.9	ER51	Kocuria rosea	MK696247	100.0
ER25	Bacillus tequilensis	MH762888	100.0	ER52	Brevibacterium frigoritolerans	KY753242	99.9
ER26	Bacillus pseudomycoides	MH578628	99.6	ER53	Halomonas hydrothermalis	CP023656	99.8
ER27	Bacillus mycoides	JN215520	99.6	ER54	Microbacterium barkeri	KY882051	99.6

菌株 Strain	最相近菌种 The closest strain	登录号 Login number	相似性 Similarity/%	菌株 Strain	最相近菌种 The closest strain	登录号 Login number	相似性 Similarity/%
ER1	Bacillus proteolyticus	NR157735	99.9	ER28	Bacillus paramycoides	MT124553	99.72
ER2	Bacillus methylotrophicus	KC790266	99.9	ER29	Pseudomonas plecoglossicida	MT124554	100.0
ER3	Bacillus toyoneusis	NR121761	100.0	ER30	Pseudomonas monteilii	MT124555	98.0
ER4	Bacillus licheniformis	MH842141	99.9	ER31	Pseudomonas taiwanensis	JF911384	100.0
ER5	Bacillus cereus	FJ501984	100.0	ER32	Pseudomonas aeruginosa	NR117678	100.0
ER6	Bacillus wiedmannii	MK088275	100.0	ER33	Pseudomonas fluorescens	MN256396	99.0
ER7	Bacillus amyloliquefacieus	KU291367	100.0	ER34	Pseudomonas oryzihabitans	JQ660200	100.0
ER8	Bacillus velezensis	MG996871	100.0	ER35	Pseudomonas psychrotolerans	KY882047	99.9
ER9	Bacillus siamensis	MN166187	100.0	ER36	Pseudomonas stutzeri	FJ768951	99.9
ER10	Bacillus subtilis	MK860021	100.0	ER37	Proteus mirabilis	MK332376	100.0
ER11	Bacillus anthracis	CP031642	99.9	ER38	Micrococcus luteus	MH045862	100.0
ER12	Bacillus aryabhattai	MK859946	100.0	ER39	Stenotrophomonas maltophilia	MH000623	100.0
ER13	Bacillus vallismortis	NR024696	100.0	ER40	Lysinibacillus parvibpronicapiens	NR114213	99.7
ER14	Bacillus oryzaecorticis	KU877642	99.9	ER41	Brachybacterium paraconglomeratum	NR025502	100.0
ER15	Bacillus aquimaris	KF358255	99.8	ER42	Agrobacterium tumefaciens	MN094859	99.9
ER16	Bacillus marisflavi	KX495310	99.8	ER43	Lysinibacillus fusiformis	KP980567	99.9
ER17	Bacillus sonerensis	MK860002	100.0	ER44	Enterobacter cloacae	KM538690	99.9
ER18	Bacillus thuringiensis	KJ769222	99.5	ER45	Enterobacter bugandensis	CP039453	99.8
ER19	Bacillus safensis	MK859936	100.0	ER46	Enterobacter hormaechei	CP027142	100.0
ER20	Bacillus simplex	KF818647	99.9	ER47	Enterobacter ludwigii	NR042349	99.9
ER21	Bacillus firmus	KP165527	99.9	ER48	Enterobacter xiangfangensis	MH200641	99.5
ER22	Bacillus altitudinis	KU663665	100.0	ER49	Enterobacter soli	NR117547	99.3
ER23	Bacillus flexus	KR999917	100.0	ER50	Terribacillus goriensis	CP008876	100.0
ER24	Bacillus horikoshii	CP020880	99.9	ER51	Kocuria rosea	MK696247	100.0
ER25	Bacillus tequilensis	MH762888	100.0	ER52	Brevibacterium frigoritolerans	KY753242	99.9
ER26	Bacillus pseudomycoides	MH578628	99.6	ER53	Halomonas hydrothermalis	CP023656	99.8
ER27	Bacillus mycoides	JN215520	99.6	ER54	Microbacterium barkeri	KY882051	99.6

菌株 Strain	产IAA量 IAA yield/（mg·L^-1）	菌株 Strain	产IAA量 IAA yield/（mg·L^-1）	菌株 Strain	产IAA量 IAA yield/（mg·L^-1）	菌株 Strain	产IAA量 IAA yield/（mg·L^-1）
ER1	3.94±0.45r	ER31	11.24±0.18gh	ER18	6.52±0.26nop	ER44	27.29±1.86a
ER2	9.22±0.29ij	ER32	5.21±0.22pqr	ER19	5.98±0.35opq	ER45	25.93±0.54a
ER4	4.76±0.20qr	ER35	8.26±0.13jkl	ER20	7.60±0.08klmn	ER46	11.43±0.35fg
ER5	5.91±0.19opq	ER36	10.74±0.11gh	ER21	8.64±0.18ijk	ER48	22.02±0.89b
ER6	7.49±0.42klmn	ER37	13.89±0.24de	ER24	6.72±0.21mno	ER49	27.35±0.20a
ER7	7.63±0.33klmn	ER38	4.12±0.33r	ER25	5.13±0.09pqr	ER50	6.96±0.50lmno
ER8	5.19±0.34pqr	ER40	7.30±0.22klmno	ER27	8.08±0.12jklm	ER51	16.20±0.21c
ER9	2.15±0.14s	ER41	16.05±0.14c	ER28	12.73±0.14ef	ER52	15.29±0.24cd
ER10	15.64±0.25c	ER42	3.89±0.27r	ER29	7.50±0.69klmn	ER53	11.61±0.34fg
ER12	13.8±0.94e	ER43	7.59±0.22klmn	ER30	7.01±0.15lmno	ER54	9.93±0.23hi
ER17	6.37±0.25nop

菌株 Strain	产IAA量 IAA yield/（mg·L^-1）	菌株 Strain	产IAA量 IAA yield/（mg·L^-1）	菌株 Strain	产IAA量 IAA yield/（mg·L^-1）	菌株 Strain	产IAA量 IAA yield/（mg·L^-1）
ER1	3.94±0.45r	ER31	11.24±0.18gh	ER18	6.52±0.26nop	ER44	27.29±1.86a
ER2	9.22±0.29ij	ER32	5.21±0.22pqr	ER19	5.98±0.35opq	ER45	25.93±0.54a
ER4	4.76±0.20qr	ER35	8.26±0.13jkl	ER20	7.60±0.08klmn	ER46	11.43±0.35fg
ER5	5.91±0.19opq	ER36	10.74±0.11gh	ER21	8.64±0.18ijk	ER48	22.02±0.89b
ER6	7.49±0.42klmn	ER37	13.89±0.24de	ER24	6.72±0.21mno	ER49	27.35±0.20a
ER7	7.63±0.33klmn	ER38	4.12±0.33r	ER25	5.13±0.09pqr	ER50	6.96±0.50lmno
ER8	5.19±0.34pqr	ER40	7.30±0.22klmno	ER27	8.08±0.12jklm	ER51	16.20±0.21c
ER9	2.15±0.14s	ER41	16.05±0.14c	ER28	12.73±0.14ef	ER52	15.29±0.24cd
ER10	15.64±0.25c	ER42	3.89±0.27r	ER29	7.50±0.69klmn	ER53	11.61±0.34fg
ER12	13.8±0.94e	ER43	7.59±0.22klmn	ER30	7.01±0.15lmno	ER54	9.93±0.23hi
ER17	6.37±0.25nop

菌株 Strain	D/d	溶磷量P-solubilizing capability /（mg·L^-1）	pH	菌株 Strain	D/d	溶磷量P-solubilizing capability /（mg·L^-1）	pH	菌株 Strain	D/d	溶磷量P-solubilizing capability /（mg·L^-1）	pH
ER4	+	36.56±0.58k	5.07±0.07ef	ER35	+++	23.25±1.53n	6.35±0.23a	ER24	+	19.03±0.91op	4.54±0.29gh
ER5	++	60.50±1.16i	5.49±0.07bcd	ER36	+	82.95±0.98ef	5.69±0.10bc	ER26	++	59.38±1.03i	5.48±0.17bcde
ER6	+++	34.75±0.91kl	5.88±0.14b	ER37	+	73.67±0.77g	5.49±0.15bcd	ER29	++	79.52±1.74f	79.52±1.74f
ER7	+	7.25±0.82a	6.47±0.06a	ER39	++	54.65±1.04j	4.78±0.21fg	ER32	+++	133.65±0.64a	133.65±0.64a
ER9	+++	120.52±1.49b	4.39±0.10gh	ER41	++	84.20±0.58e	5.75±0.11bc	ER33	++	67.27±1.41h	5.65±0.21bc
ER10	++	59.85±1.40i	5.41±0.09cde	ER42	++	59.29±2.32i	4.37±0.1gh	ER48	+	66.25±0.77h	5.48±0.12bcde
ER12	++	15.92±0.64p	6.34±0.07a	ER43	++	29.14±1.21m	5.17±0.08def	ER50	+	6.32±0.28q	5.64±0.13bc
ER15	+	103.27±0.66d	4.42±0.06gh	ER45	+	17.36±0.49p	6.34±0.06a	ER51	+++	112.91±0.61c	4.58±0.18gh
ER17	+++	7.94±1.74a	5.46±0.09bcde	ER46	+	22.69±0.35no	5.34±0.12cde	ER53	++	32.20±0.77lm	6.46±0.09a
ER20	+	60.73±2.02i	4.64±0.07gh	ER47	+	53.86±1.53j	4.34±0.04h