Biocontrol Characteristics of Bacillus atrophaeus CNY01 and Its Salt-resistant and Growth-promoting Effect on Maize Seedling

doi:10.13560/j.cnki.biotech.bull.1985.2023-1174

Abstract

Abstract:

【Objective】A rhizobacterium with salt-resistant and growth-promoting effects was screened from the saline-alkali soil in Yellow River Delta to prepare an agent for alleviating high-salt stress of maize seedlings. 【Method】The salt-resistant and growth-promoting functions of the strain were identified on functional mediums, and the species of the strain was determined by 16S rDNA sequence and whole genome analysis. The strain was applied to maize rhizosphere to verify the salt-resistant and growth-promoting effects of the strain on maize seedlings.【Result】A strain Bacillus atrophaeus CNY01 was screened from the saline-alkali soil of Yellow River Delta. Strain CNY01 grew normally under the condition of 0-16% NaCl and pH 5-8. Strain CNY01 had the ability of degrading protein and dissolving potassium, and inhibited the growth of plant pathogens such as Ralstonia solanacearum and Fusarium moniliforme. The application of strain CNY01 promoted the growth of maize seedlings under high-salt condition and alleviated the stress of high salt on maize seedlings. When strain CNY01 was applied to maize seedlings in nutrient solution containing 100 mmol/L NaCl for 8 d, the physiological plant height, root length, and fresh weight of maize seedlings increased by 38.80%（P<0.01）, 23.73%, and 28.19%（P<0.01）, respectively. When strain CNY01 was applied to maize seedlings in pot experiments adding 100 mmol/L NaCl for 28 d, the physiological plant height, the aboveground fresh weight, and the underground fresh weight of maize seedlings increased by 10.84%, 41.87%（P<0.01）, and 23.29%, respectively. Genes that maintain cell osmotic pressure and synthesize stress proteins to alleviate high salt stress were also predicted in the genome sequence of this strain.【Conclusion】A strain of B. atrophaeus CNY01 is screened from saline-alkali soil of Yellow River Delta, which has the function of preventing disease and promoting growth, and has significant salt-resistant and promoting growth effect on maize plants. Meanwhile, strain CNY01 contains genes related to salt-resistant and growth-promoting. B. atrophaeus CNY01 is an important strain resource.

Key words: maize, plant growth-promoting rhizobacteria, Bacillus atrophaeus, salt-resistant and growth-promoting, whole-genome sequencing

SUN Ya-nan, WANG Chun-xue, WANG Xin, DU Bing-hai, LIU Kai, WANG Cheng-qiang. Biocontrol Characteristics of Bacillus atrophaeus CNY01 and Its Salt-resistant and Growth-promoting Effect on Maize Seedling[J]. Biotechnology Bulletin, 2024, 40(5): 248-260.

Figures/Tables 10

References 33

[1]	王佳丽, 黄贤金, 钟太洋, 等. 盐碱地可持续利用研究综述[J]. 地理学报, 2011, 66(5): 673-684.
	Wang JL, Huang XJ, Zhong TY, et al. Review on sustainable utilization of salt-affected land[J]. Acta Geogr Sin, 2011, 66(5): 673-684. doi: 10.11821/xb201105010
[2]	魏博娴. 中国盐碱土的分布与成因分析[J]. 水土保持应用技术, 2012(6): 27-28.
	Wei BX. Distribution and cause analysis of saline-alkali soil in China[J]. Technol Soil Water Conserv, 2012(6): 27-28.
[3]	赵娇. 基于菌群改善促进盐碱地耐受性植物生长的研究[D]. 济南: 山东大学, 2020.
	Zhao J. Study on the improvement of flora to promote thegrowth of tolerant plants in saline-alkali land[D]. Jinan: Shandong University, 2020.
[4]	王文飞. 萎缩芽孢杆菌WU-9与辣椒互作缓解盐胁迫机制及菌株相关功能基因解析[D]. 石河子: 石河子大学, 2022.
	Wang WF. Mechanism of relieving salt stress by interaction between Bacillus Atrophyta WU-9 and pepper and analysis of related functional genes of the strain[D]. Shihezi: Shihezi University, 2022.
[5]	郭英. 生防芽孢杆菌对作物耐盐性的影响及其抗盐机理的研究[D]. 济南: 山东师范大学, 2009.
	Guo Y. Effects and mechanism of biocontrol strain of Bacillus subtilis on salt tolerance of plants[D]. Jinan: Shandong Normal University, 2009.
[6]	庞亚琴, 任彩婷, 徐秋曼. 解淀粉芽孢杆菌HM618对镉胁迫下小麦幼苗生长的影响[J]. 天津师范大学学报: 自然科学版, 2018, 38(4): 55-59.
	Pang YQ, Ren CT, Xu QM. Effects of Bacillus amyloliquefaciens HM618 on the growth of wheat seedlings under cadmium stress[J]. J Tianjin Norm Univ Nat Sci Ed, 2018, 38(4): 55-59.
[7]	Sella SRBR, Vandenberghe LPS, Soccol CR. Bacillus atrophaeus: main characteristics and biotechnological applications - a review[J]. Crit Rev Biotechnol, 2015, 35(4): 533-545. doi: 10.3109/07388551.2014.922915 pmid: 24963702
[8]	Ma JJ, Wang CQ, Wang HD, et al. Analysis of the complete genome sequence of Bacillus atrophaeus GQJK17 reveals its biocontrol characteristics as a plant growth-promoting rhizobacterium[J]. Biomed Res Int, 2018, 2018: 9473542.
[9]	Gordon RE, Smith NR. Aerobic sporeforming bacteria capable of growth at high temperatures[J]. J Bacteriol, 1949, 58(3): 327-341. doi: 10.1128/jb.58.3.327-341.1949 pmid: 16561790
[10]	Nakamura LK. Taxonomic relationship of black-pigmented Bacillus subtilis strains and a proposal for Bacillus atrophaeus sp. nov.[J]. International Journal of Systematic and Evolutionary Microbiology, 1989, 39(3):295-300.
[11]	Hou YL, Zeng WZ, Ao C, et al. Bacillus atrophaeus WZYH01 and Planococcus soli WZYH02 improve salt tolerance of maize(Zea mays L.) in saline soil[J]. Front Plant Sci, 2022, 13: 891372.
[12]	韦廷舟, 文怡, 王超, 等. 一株产IAA芽孢杆菌ST37对油菜的耐盐促生作用[J]. 江苏农业科学, 2023, 51(6): 210-215.
	Wei TZ, Wen Y, Wang C, et al. Effect of an IAA-producing Bacillus ST37 on salt tolerance and growth promotion of rape[J]. Jiangsu Agric Sci, 2023, 51(6): 210-215.
[13]	郇惠杰, 钟泓波, 雷芬芬, 等. 产蛋白酶海洋细菌的筛选、鉴定及发酵培养基的研究[J]. 食品工业科技, 2013, 34(24): 181-185.
	Huan HJ, Zhong HB, Lei FF, et al. Study on isolation and identification of protease-producing marine bacteria and optimization of fermentation medium[J]. Sci Technol Food Ind, 2013, 34(24): 181-185.
[14]	张祥胜. 发酵液有效磷含量测定方法研究[J]. 湖州职业技术学院学报, 2008, 6(3): 1-3.
	Zhang XS. A study of factors affecting the determined value by Mo-Sn-vc method of organic phosphobacteria[J]. J Huzhou Vocat Technol Coll, 2008, 6(3): 1-3.
[15]	李浩. 玉米、黄瓜根际促生菌组合优化及基因组测序[D]. 泰安: 山东农业大学, 2019.
	Li H. Optimization of rhizosphere growth-promoting bacteria combination and genome sequencing of maize and cucumber[D]. Tai'an: Shandong Agricultural University, 2019.
[16]	马锦锦. 枸杞根际促生细菌筛选、培养基优化及基因组测序[D]. 泰安: 山东农业大学, 2018.
	Ma JJ. Screening, Medium optimization and genome sequencing of PGPR from the rhizosphere of Lycium barbarum L.[D]. Tai'an: Shandong Agricultural University, 2018.
[17]	侯贞. 烟草根际促生细菌的筛选鉴定及发酵培养基优化[D]. 泰安: 山东农业大学, 2015.
	Hou Z. Screening and identification of tobacco rhizosphere growth-promoting bacteria and optimization of fermentation medium[D]. Tai'an: Shandong Agricultural University, 2015.
[18]	杨亚男. 番茄根际促生菌的筛选及其培养基优化[D]. 泰安: 山东农业大学, 2017.
	Yang YN. Screening of tomato rhizosphere growth-promoting bacteria and optimization of its culture medium[D]. Tai'an: Shandong Agricultural University, 2017.
[19]	Schubert M, Lindgreen S, Orlando L. AdapterRemoval v2: rapid adapter trimming, identification, and read merging[J]. BMC Res Notes, 2016, 9: 88. doi: 10.1186/s13104-016-1900-2 pmid: 26868221
[20]	Luo RB, Liu BH, Xie YL, et al. SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler[J]. GigaScience, 2012, 1(1): 18.
[21]	Chin CS, Peluso P, Sedlazeck FJ, et al. Phased diploid genome assembly with single-molecule real-time sequencing[J]. Nat Methods, 2016, 13(12): 1050-1054. doi: 10.1038/NMETH.4035
[22]	Koren S, Walenz BP, Berlin K, et al. Canu: scalable and accurate long-read assembly via adaptive k-mer weighting and repeat separation[J]. Genome Res, 2017, 27(5): 722-736.
[23]	Walker BJ, Abeel T, Shea T, et al. Pilon: an integrated tool for comprehensive microbial variant detection and genome assembly improvement[J]. PLoS One, 2014, 9(11): e112963.
[24]	王欣. 植物根际促生细菌CNY01和BY2G20的耐盐功能及其对玉米的抗盐促生研究[D]. 泰安: 山东农业大学, 2021.
	Wang X, The Salt Tolerance Function of Plant Growth-promoting Rhizobacteria CNY01 and BY2G20 and Their Anti-salt Promotion for Corn Growth[D]. Tai'an: Shandong Agricultural University, 2021.
[25]	索雲凯, 刘丽红, 张雷, 等. 解钾菌解钾作用研究进展[J]. 当代化工, 2021, 50(4): 924-929.
	Suo YK, Liu LH, Zhang L, et al. Research progress of potassium solubilization by potassium solubilizing bacteria[J]. Contemp Chem Ind, 2021, 50(4): 924-929.
[26]	Ayaz M, Ali Q, Farzand A, et al. Nematicidal volatiles from Bacillus atrophaeus GBSC56 promote growth and stimulate induced systemic resistance in tomato against Meloidogyne incognita[J]. Int J Mol Sci, 2021, 22(9): 5049.
[27]	刘思靖. 萎缩芽孢杆菌次级代谢产物的研究进展[J]. 现代化工, 2019, 39(12): 48-51.
	Liu SJ. Advances in secondary metabolites from Bacillus atrophaeus[J]. Mod Chem Ind, 2019, 39(12): 48-51.
[28]	戴宝, 扈进冬, 尹姗姗, 等. 响应面法优化萎缩芽孢杆菌BsR05发酵培养基[J]. 山东科学, 2017, 30(4): 31-37. doi: 10.3976/j.issn.1002-4026.2017.04.006
	Dai B, Hu JD, Yin SS, et al. Optimization of fermentation medium composition for Bacillus atrophaeus BsR05 by response surface method[J]. Shandong Sci, 2017, 30(4): 31-37.
[29]	王敏. 萎缩芽孢杆菌XW2对苹果树腐烂病的防效评价[D]. 乌鲁木齐: 新疆农业大学, 2020.
	Wang M. Evaluation on control effect of Bacillus atrophyta XW2 on apple tree rot[D]. Urumqi: Xinjiang Agricultural University, 2020.
[30]	胡玉婕, 朱秀玲, 丁延芹, 等. 芽孢杆菌的耐盐促生机制研究进展[J]. 生物技术通报, 2020, 36(9): 64-74. doi: 10.13560/j.cnki.biotech.bull.1985.2020-0746
	Hu YJ, Zhu XL, Ding YQ, et al. Research progress on salt tolerance and growth-promoting mechanism of Bacillus[J]. Biotechnol Bull, 2020, 36(9): 64-74.
[31]	Karim R, Bouchra B, Fatima G, et al. Plant NHX antiporters: from function to biotechnological application, with case study[J]. Curr Protein Pept Sci, 2021, 22(1): 60-73.
[32]	赵东晓, 董亚茹, 孙景诗, 等. NaCl胁迫对胡麻种子萌发、幼苗生长及Na⁺/H⁺逆向转运蛋白基因表达的影响[J]. 山东农业科学, 2020, 52(7): 40-45.
	Zhao DX, Dong YR, Sun JS, et al. Effects of NaCl stress on seed germination, seedling growth and Na⁺/H⁺ antiporter protein gene expression of Linum usitatissimum L[J]. Shandong Agric Sci, 2020, 52(7): 40-45.
[33]	张傲洁, 李青云, 宋文红, 等. 基于苯酚降解的粪产碱杆菌Alcaligenes faecalis JF101的全基因组分析[J]. 生物技术通报, 2023, 39(10): 292-303. doi: 10.13560/j.cnki.biotech.bull.1985.2023-0281
	Zhang AJ, Li QY, Song WH, et al. Whole genome sequencing analysis of a phenol-degrading strain Alcaligenes faecalis JF101[J]. Biotechnol Bull, 2023, 39(10): 292-303.

Genomic information	CNY01	GQJK17	BA59	PENSV20	1942	SRCM101359	NS2	NX-12	MBLB1156
Genome Size/Mb	4.14	4.33	4.26	4.15	4.17	4.18	4.26	4.16	4.15
G+C content/%	43.50	43.30	43.10	43.50	43.20	43.30	43.30	43.30	43.40
CDSs（with protein）	3950	4045	3889	3932	3995	3923	4018	3946	3870
Pseudo genes number	146	161	383	157	152	157	159	158	232
rRNA genes number	24	24	26	24	21	24	24	24	24
tRNA genes number	82	84	84	83	75	82	82	82	82
The source of isolation	Soil （China）	Rhizophere （China）	Cotton root （China）	Soil （Canada）	-	Food （South Korea）	-	Cotton rhizosphere soil	Fermented to fu

Genomic information	CNY01	GQJK17	BA59	PENSV20	1942	SRCM101359	NS2	NX-12	MBLB1156
Genome Size/Mb	4.14	4.33	4.26	4.15	4.17	4.18	4.26	4.16	4.15
G+C content/%	43.50	43.30	43.10	43.50	43.20	43.30	43.30	43.30	43.40
CDSs（with protein）	3950	4045	3889	3932	3995	3923	4018	3946	3870
Pseudo genes number	146	161	383	157	152	157	159	158	232
rRNA genes number	24	24	26	24	21	24	24	24	24
tRNA genes number	82	84	84	83	75	82	82	82	82
The source of isolation	Soil （China）	Rhizophere （China）	Cotton root （China）	Soil （Canada）	-	Food （South Korea）	-	Cotton rhizosphere soil	Fermented to fu

Name of strain	Bacillus haynesii	Bacillus sonorensis	Bacillus siamensis	Bacillus licheniformis	Bacillus atrophaeus	Bacillus paralicheniformis	CNY01
Bacillus haynesii	*	81.18	71.86	95.14	72.29	95.14	72.23
Bacillus sonorensis	80.85	*	71.84	80.97	72.47	80.72	72.38
Bacillus siamensis	72.04	71.92	*	71.93	76.64	71.84	76.73
Bacillus licheniformis	95.40	81.42	71.94	*	72.45	94.38	72.43
Bacillus atrophaeus	72.28	72.57	76.66	72.30	*	72.32	98.06
Bacillus paralicheniformis	95.03	81.00	71.98	94.02	72.25	*	72.13
CNY01	72.32	72.63	76.89	72.33	98.36	72.34	*

Name of strain	Bacillus haynesii	Bacillus sonorensis	Bacillus siamensis	Bacillus licheniformis	Bacillus atrophaeus	Bacillus paralicheniformis	CNY01
Bacillus haynesii	*	81.18	71.86	95.14	72.29	95.14	72.23
Bacillus sonorensis	80.85	*	71.84	80.97	72.47	80.72	72.38
Bacillus siamensis	72.04	71.92	*	71.93	76.64	71.84	76.73
Bacillus licheniformis	95.40	81.42	71.94	*	72.45	94.38	72.43
Bacillus atrophaeus	72.28	72.57	76.66	72.30	*	72.32	98.06
Bacillus paralicheniformis	95.03	81.00	71.98	94.02	72.25	*	72.13
CNY01	72.32	72.63	76.89	72.33	98.36	72.34	*

位置Location	长度Length/bp	基因Gene	产物Product
KCX77_05795	453	nhaC	Na⁺/H⁺ antiporter NhaC
KCX77_09890	429	/	Na⁺/H⁺ antiporter NhaC family protein
KCX77_12210	468	nhaC	Na⁺/H⁺ antiporter NhaC
KCX77_15895	806	/	Na⁺/H⁺ antiporter subunit A
KCX77_15910	493	/	Na⁺/H⁺ antiporter subunit D
KCX77_15915	158	/	Na⁺/H⁺ antiporter subunit E
KCX77_15925	124	/	Na⁺/H⁺ antiporter subunit G
KCX77_16910	671	/	Na⁺/H⁺ antiporter
KCX77_01700	418	opuAA	Glycine/proline betaine ABC transporter
KCX77_01705	282	opuAB	Glycine/proline betaine ABC transporter permease subunit OpuAB
KCX77_01805	303	/	Proline dehydrogenase
KCX77_01815	504	putP	Sodium/proline symporter PutP
KCX77_04225	492	putP	Sodium/proline symporter PutP
KCX77_09365	564	proS	Proline--tRNA ligase
KCX77_16595	302	/	Proline dehydrogenase
KCX77_17080	381	/	Betaine/proline/choline family ABC transporter ATP-binding protein
KCX77_00320	205	/	General stress protein Ctc
KCX77_04635	115	/	General stress protein
KCX77_15790	130	yugI	General stress protein 13
KCX77_19470	547	katX	Catalase KatX
KCX77_02490	273	/	Manganese catalase family protein
KCX77_05335	483	katA	Catalase KatA
KCX77_12565	285	/	Manganese catalase family protein
KCX77_08110	302	/	Manganese catalase family protein
KCX77_19655	685	/	Catalase
KCX77_07800	449	/	TrkH family potassium uptake protein
KCX77_08320	221	ktrC	Ktr system potassium transporter KtrC
KCX77_15595	222	/	TrkA family potassium uptake protein
KCX77_15600	445	/	TrkH family potassium uptake protein
KCX77_15755	328	/	Potassium channel family protein
KCX77_19400	221	/	TrkA family potassium uptake protein
KCX77_17200	394	/	Phosphoglycerate kinase
KCX77_14540	585	pyk	Pyruvate kinase