Identification of Bacillus cereus YT2-1C with High Indoleacetic Acid Yield and Its Growth-promoting Effect

doi:10.13560/j.cnki.biotech.bull.1985.2024-1076

Abstract

Abstract:

Objective The growth promotion and bacteriostatic potential of strain YT2-1C with high yield of indoleacetic acid was analyzed to provide a basis for its subsequent development and utilization. Method The strain was identified by morphological characteristics, 16S rRNA and 31 house-keeping genes. The IAA yield of strain was quantitatively determined. The potential of the strain in iron production, phosphorus solubilization, potassium solubilization, protease production, nitrogen fixation and bacteriostasis were analyzed. The growth-promoting effect of the strain was further verified by seedling breeding and field experiments. The functional genes related to growth-promoting were preliminarily analyzed from the genome level. Result The strain was identified as Bacillus cereus. The IAA yield of this strain reached 193 μg/ml. It had the ability of producing iron carrier, dissolving phosphorus, producing protease, and inhibiting bacteria. The results from breeding experiment showed that the plant height, chlorophyll, stem diameter and fresh weight of cucumber, pepper and tomato significantly increased by inoculation of YT2-1C strain. There was still good growth-promoting effect after cucumber plants transplanted to the field. Whole genome sequencing analysis showed that strain YT2-1C had 54 genes related to growth promotion and 5 genes related to non-ribosomal peptide synthesis. Conclusion Bacillus cereus YT2-1C is a strain that has the function of promoting growth and preventing disease. It has significant growth-promoting effect on cucumber, pepper and tomato, and contains many genes related to growth-promoting and disease-preventing, which lays a foundation for the development of its microbial microbial agent.

Key words: growth-promoting bacteria, Bacillus cereus, strain YT2-1C, IAA, seedling substrate, whole genome sequencing

ZHANG Hui, LU Wen-cai, WANG Dong, LIU Qian, MA Lian-jie. Identification of Bacillus cereus YT2-1C with High Indoleacetic Acid Yield and Its Growth-promoting Effect[J]. Biotechnology Bulletin, 2025, 41(5): 300-309.

Figures/Tables 10

References 37

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作物 Crop	处理 Treatment	株高 Plant height （cm）	叶绿素Chlorophyll （SPAD）	茎粗 Stem size （mm）	鲜重 Fresh weight （g）
辣椒	CK	6.20a	33.28a	2.02a	0.86a
辣椒	YT2-1C	10.2b	37.28b	2.64b	1.33b
番茄	CK	8.32a	2.84a	2.02a	1.78a
番茄	YT2-1C	19.12b	5.21b	2.64b	7.49b

作物 Crop	处理 Treatment	株高 Plant height （cm）	叶绿素Chlorophyll （SPAD）	茎粗 Stem size （mm）	鲜重 Fresh weight （g）
辣椒	CK	6.20a	33.28a	2.02a	0.86a
辣椒	YT2-1C	10.2b	37.28b	2.64b	1.33b
番茄	CK	8.32a	2.84a	2.02a	1.78a
番茄	YT2-1C	19.12b	5.21b	2.64b	7.49b

功能 Function	基因 Gene	产物 Product
IAA合成 IAA synthesis	trpA	色氨酸合成酶ɑ亚基
	trpB	色氨酸合成酶β亚基
	trpC	吲哚-3-甘油磷酸合成酶
	trpD	邻氨基苯甲酸磷酸核糖基转移酶
	trpE	邻氨基苯甲酸合酶
	trpG	邻氨基苯甲酸合酶
	ipdC	吲哚3-丙酮酸脱羧酶
	aroA	3-磷酸莽草酸1-羧乙烯基转移酶
	sucA	2-氧戊二酸脱氢酶E1组分
	kynB	芳基甲酰胺酶
	kynU	尿氨酸酶
	atoB	乙酰辅酶a-c-乙酰转移酶
	amiE	酰胺酶家族
	aofH	黄素单胺氧化酶
铁载体 Siderophore	entA	2，3-二氢-2，3-二羟基苯甲酸脱氢酶
	entB	异分支酸酶家族蛋白
	entC	异氯酸合成酶DhbC
	entE	（2，3-二羟基苯甲酰）腺苷酸合酶
	menF	同氯酸酯合成酶
	mbtH	MbtH家族蛋白
	nuoA	NADH-醌氧化还原酶亚基NuoA
	nuoB	NADH-醌氧化还原酶亚基NuoB
	nuoC	NADH-醌氧化还原酶亚基C
	nuoD	NADH-醌氧化还原酶亚基NuoD
	nuoI	NADH-醌氧化还原酶亚基nui
	nuoJ	NADH-醌氧化还原酶亚基J
	nuoH	NADH-醌氧化还原酶亚基NuoH
	nuoK	NADH-醌氧化还原酶亚基NuoK
	nuoL	NADH-醌氧化还原酶亚基L
	nuoN	NADH-醌氧化还原酶亚基NuoN
	nuoM	NADH-醌氧化还原酶亚基M
	bfr	类铁蛋白结构域含蛋白
	ftnA	铁蛋白
溶磷 Phosphorus solubilization	patA	转氨酶A
	pstA	磷酸ABC转运蛋白渗透酶
	pstC	磷酸ABC转运蛋白渗透酶
	pyk	丙酮酸激酶
	pyc	丙酮酸羧化酶
	ackA	醋酸激酶
	gltA	柠檬酸合酶
	buk	丁酸激酶
	aroK	莽草激酶
产蛋白酶 Proteinase production	aprE	S8家族肽酶
	nprE	M4家族金属肽酶
	nprB	中性蛋白酶B前体
	vpr	S8家族丝氨酸肽酶
	mprF	双功能赖氨酸磷脂酰甘油翻转酶/合成酶
固氮 Nitrogen fixation	glnA	谷氨酸-氨连接酶
	gltB	谷氨酸合酶
	narG	硝酸盐还原酶
	narH	硝酸盐还原酶
	narI	亚硝酸盐氧化还原酶
	nirA	铁氧还蛋白-亚硝酸盐还原酶
	cynT	碳酸酐酶
非核糖体肽 Non-ribosomal peptides	tycC	非核糖体肽合成酶
	dhbF	非核糖体肽合成酶
	mcyA	非核糖体肽合成酶
	ituB	非核糖体肽合成酶
	bacA	非核糖体肽合成酶

功能 Function	基因 Gene	产物 Product
IAA合成 IAA synthesis	trpA	色氨酸合成酶ɑ亚基
	trpB	色氨酸合成酶β亚基
	trpC	吲哚-3-甘油磷酸合成酶
	trpD	邻氨基苯甲酸磷酸核糖基转移酶
	trpE	邻氨基苯甲酸合酶
	trpG	邻氨基苯甲酸合酶
	ipdC	吲哚3-丙酮酸脱羧酶
	aroA	3-磷酸莽草酸1-羧乙烯基转移酶
	sucA	2-氧戊二酸脱氢酶E1组分
	kynB	芳基甲酰胺酶
	kynU	尿氨酸酶
	atoB	乙酰辅酶a-c-乙酰转移酶
	amiE	酰胺酶家族
	aofH	黄素单胺氧化酶
铁载体 Siderophore	entA	2，3-二氢-2，3-二羟基苯甲酸脱氢酶
	entB	异分支酸酶家族蛋白
	entC	异氯酸合成酶DhbC
	entE	（2，3-二羟基苯甲酰）腺苷酸合酶
	menF	同氯酸酯合成酶
	mbtH	MbtH家族蛋白
	nuoA	NADH-醌氧化还原酶亚基NuoA
	nuoB	NADH-醌氧化还原酶亚基NuoB
	nuoC	NADH-醌氧化还原酶亚基C
	nuoD	NADH-醌氧化还原酶亚基NuoD
	nuoI	NADH-醌氧化还原酶亚基nui
	nuoJ	NADH-醌氧化还原酶亚基J
	nuoH	NADH-醌氧化还原酶亚基NuoH
	nuoK	NADH-醌氧化还原酶亚基NuoK
	nuoL	NADH-醌氧化还原酶亚基L
	nuoN	NADH-醌氧化还原酶亚基NuoN
	nuoM	NADH-醌氧化还原酶亚基M
	bfr	类铁蛋白结构域含蛋白
	ftnA	铁蛋白
溶磷 Phosphorus solubilization	patA	转氨酶A
	pstA	磷酸ABC转运蛋白渗透酶
	pstC	磷酸ABC转运蛋白渗透酶
	pyk	丙酮酸激酶
	pyc	丙酮酸羧化酶
	ackA	醋酸激酶
	gltA	柠檬酸合酶
	buk	丁酸激酶
	aroK	莽草激酶
产蛋白酶 Proteinase production	aprE	S8家族肽酶
	nprE	M4家族金属肽酶
	nprB	中性蛋白酶B前体
	vpr	S8家族丝氨酸肽酶
	mprF	双功能赖氨酸磷脂酰甘油翻转酶/合成酶
固氮 Nitrogen fixation	glnA	谷氨酸-氨连接酶
	gltB	谷氨酸合酶
	narG	硝酸盐还原酶
	narH	硝酸盐还原酶
	narI	亚硝酸盐氧化还原酶
	nirA	铁氧还蛋白-亚硝酸盐还原酶
	cynT	碳酸酐酶
非核糖体肽 Non-ribosomal peptides	tycC	非核糖体肽合成酶
	dhbF	非核糖体肽合成酶
	mcyA	非核糖体肽合成酶
	ituB	非核糖体肽合成酶
	bacA	非核糖体肽合成酶

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[7]	CHANG Lu-yin, WANG Zhong-hua, LI Feng-min, GAO Zi-yuan, ZHANG Hui-hong, WANG Yi, LI Fang, HAN Yan-lai, JIANG Ying. Screening Multi-functional Rhizobacteria from Maize Rhizosphere and Their Ehancing Effects on Winter Wheat-Summer Maize Rotation System [J]. Biotechnology Bulletin, 2024, 40(1): 231-242.
[8]	WANG Teng-hui, GE Wen-dong, LUO Ya-fang, FAN Zhen-yu, WANG Yu-shu. Gene Mapping of Kale White Leaves Based on Whole Genome Re-sequencing of Extreme Mixed Pool（BSA） [J]. Biotechnology Bulletin, 2023, 39(9): 176-182.
[9]	FANG Lan, LI Yan-yan, JIANG Jian-wei, CHENG Sheng, SUN Zheng-xiang, ZHOU Yi. Isolation, Identification and Growth-promoting Characteristics of Endohyphal Bacterium 7-2H from Endophytic Fungi of Spiranthes sinensis [J]. Biotechnology Bulletin, 2023, 39(8): 272-282.
[10]	GUO Shao-hua, MAO Hui-li, LIU Zheng-quan, FU Mei-yuan, ZHAO Ping-yuan, MA Wen-bo, LI Xu-dong, GUAN Jian-yi. Whole Genome Sequencing and Comparative Genome Analysis of a Fish-derived Pathogenic Aeromonas Hydrophila Strain XDMG [J]. Biotechnology Bulletin, 2023, 39(8): 291-306.
[11]	LUO Yi, ZHANG Li-juan, HUANG Wei, WANG Ning, Wuerlika MAITIHASEM, SHI Chong, WANG Wei. Identification of a Uranium-resistant Strain and Its Growth-promoting Properties [J]. Biotechnology Bulletin, 2023, 39(5): 286-296.
[12]	ZHANG Zhi-xia, LI Tian-pei, ZENG Hong, ZHU Xi-xian, YANG Tian-xiong, MA Si-nan, HUANG Lei. Genome Sequencing and Bioinformatics Analysis of Gelidibacter sp. PG-2 [J]. Biotechnology Bulletin, 2023, 39(3): 290-300.
[13]	HE Meng-ying, LIU Wen-bin, LIN Zhen-ming, LI Er-tong, WANG Jie, JIN Xiao-bao. Whole Genome Sequencing and Analysis of an Anti Gram-positive Bacterium Gordonia WA4-43 [J]. Biotechnology Bulletin, 2023, 39(2): 232-242.
[14]	CHE Yong-mei, LIU Guang-chao, GUO Yan-ping, YE Qing, ZHAO Fang-gui, LIU Xin. Preparation of Compound Halotolerant Bioinoculant and Study on Its Growth-promoting Effect [J]. Biotechnology Bulletin, 2023, 39(11): 217-225.
[15]	ZHANG Ao-jie, LI Qing-yun, SONG Wen-hong, YAN Shao-hui, TANG Ai-xing, LIU You-yan. Whole Genome Sequencing Analysis of a Phenol-degrading Strain Alcaligenes faecalis JF101 [J]. Biotechnology Bulletin, 2023, 39(10): 292-303.