Saline-Alkali Tolerant Endophytic Bacteria in Tomato: Screening, Identification and Functional Study

doi:10.11924/j.issn.1000-6850.casb2022-0940

Abstract

Abstract:

This study aimed to explore the resources of saline-alkali tolerant endophytic bacteria and to provide new strain resources for the development of endophytic fertilizer and biocontrol products. Using ‘Jinpeng No. 8’ tomato as plant material, five different types of saline-alkali environment were simulated, endophytic bacteria were isolated and identified by 16S rRNA molecular method, and then the function of bacteria were screened by plating method. A total of 14 strains of saline-alkali tolerant endophytic bacteria were obtained in this study, among which 5 strains had the ability to solve inorganic phosphorus, 2 strains had the ability to dissolve potassium, 2 strains were capable of nitrogen fixation, and 4 strains had the ability to produce auxin; 6 strains had the ability to produce iron carriers, the highest diameter ratio (D/d) was 2.95; 10 strains had the ability to produce protease, and the highest diameter ratio (D/d) was 2.97. The antagonistic results showed that 8 strains of saline-alkali tolerant endophytic bacteria could inhibit the pathogen of apple tree rot, and the inhibition rate was 13.70%-68.56%. Bacillus sp. BFG 3-3, BFG 5-1 and Streptomyces violascens BFY 2-2 showed antagonistic against 6-7 strains of common fruit and vegetable pathogens, exhibiting broad spectrum antibacterial activity. In this study, saline-alkali tolerant endophytic bacteria with the potentials of promoting plant growth and antagonism against pathogens were obtained, which provided theoretical basis for the application of endophytic bacteria in the amendment of saline-alkali soil.

Key words: saline-alkali tolerant, endophytic bacteria in tomato, isolation and cultivation, growth promoting, antagonism

LU Yuxin, ZHAO Wenjuan, BAI Yani, FENG Zhizhen, YAN Hong, QIN Tao. Saline-Alkali Tolerant Endophytic Bacteria in Tomato: Screening, Identification and Functional Study[J]. Chinese Agricultural Science Bulletin, 2023, 39(24): 50-58.

Figures/Tables 11

References 29

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序号	pH	NaCl	Na₂SO₄	Na₂CO₃
1	9	5.0%	1.0%	0
2	9	1.0%	5.0%	0
3	10	5.0%	1.0%	0
4	10	1.0%	5.0%	0
5	10	1.0%	0.5%	0.5%

序号	pH	NaCl	Na₂SO₄	Na₂CO₃
1	9	5.0%	1.0%	0
2	9	1.0%	5.0%	0
3	10	5.0%	1.0%	0
4	10	1.0%	5.0%	0
5	10	1.0%	0.5%	0.5%

培养基名称	来源或配方
蒙金娜有机磷培养基、PKO无机磷培养基、硅酸盐细菌培养基、CAS检测培养基	购买于青岛高科技工业园海博生物技术有限公司
Ashby无氮琼脂培养基(1000 mL)	葡萄糖10.0 g、K₂HPO₄ 0.2 g、K₂SO₄ 0.2 g、NaCl 0.2 g、CaCO₃ 5 g、MgSO₄·7H₂O 0.2 g、琼脂粉18.0 g
蛋白酶检测培养基	A：脱脂奶粉8 g溶于300 mL水中，115℃灭菌10 min；B：琼脂8 g加水定容至300 mL， 121℃灭菌20 min；A与B分别灭菌后混合
纤维素酶活性测定平板(1000 mL)	蛋白胨10 g、酵母粉10 g、羧甲基纤维素钠10 g、NaCl 5 g、KH₂PO₄ 1 g、琼粉脂18 g、pH 7.0
胶体状几丁质制备方法(1000 mL)	NH₄H₂PO₄ 1.0 g、KCl 0.2 g、MgSO₄·7H₂O 0.2 g、1%胶体状几丁质100 mL、琼脂粉20 g、pH 7.0

培养基名称	来源或配方
蒙金娜有机磷培养基、PKO无机磷培养基、硅酸盐细菌培养基、CAS检测培养基	购买于青岛高科技工业园海博生物技术有限公司
Ashby无氮琼脂培养基(1000 mL)	葡萄糖10.0 g、K₂HPO₄ 0.2 g、K₂SO₄ 0.2 g、NaCl 0.2 g、CaCO₃ 5 g、MgSO₄·7H₂O 0.2 g、琼脂粉18.0 g
蛋白酶检测培养基	A：脱脂奶粉8 g溶于300 mL水中，115℃灭菌10 min；B：琼脂8 g加水定容至300 mL， 121℃灭菌20 min；A与B分别灭菌后混合
纤维素酶活性测定平板(1000 mL)	蛋白胨10 g、酵母粉10 g、羧甲基纤维素钠10 g、NaCl 5 g、KH₂PO₄ 1 g、琼粉脂18 g、pH 7.0
胶体状几丁质制备方法(1000 mL)	NH₄H₂PO₄ 1.0 g、KCl 0.2 g、MgSO₄·7H₂O 0.2 g、1%胶体状几丁质100 mL、琼脂粉20 g、pH 7.0

序号	致病类型	菌名
1	苹果轮纹病	Botryosphaeria sp.（梨生囊孢壳菌）
2	黄瓜灰霉病	Botrytis cinerea（灰葡萄孢霉）
3	茄子根腐病	Alternaria sp.（链格孢霉）
4	果树根腐病	Fusarium oxysporum（尖孢镰刀菌）
5	苹果树腐烂病	Valsa sp.（黑腐皮壳菌）
6	西瓜枯萎病	Fusarium oxysporum f.sp. niveum（尖孢镰刀菌西瓜专化型）
7	猕猴桃溃疡病	Pseudomonas syringae pv. actinidiae（丁香假单孢菌猕猴桃变种）
8	番茄青枯病	Ralstonia solanacearum（青枯雷尔氏菌）