一株耐盐碱卡氏芽胞杆菌的分离及其促生功能鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2025-1367

摘要/Abstract

摘要：

目的分离、筛选耐盐碱根际促生菌，为盐碱地专用功能型微生物肥料开发和盐碱地改良提供微生物资源与理论支撑。方法采集新疆英吉沙县棉花的根际土壤，利用系列稀释法分离、筛选耐盐碱细菌；基于16S rRNA的基因序列对菌株进行分子鉴定，结合形态学和生理生化特性鉴定结果，确定菌株的分类地位；采用不同NaCl质量浓度（0-200 g/L）和不同pH值（4.0-13.0）的培养基，评估菌株的盐碱耐受性；利用多种功能鉴定培养基测定菌株的溶磷、产铁载体、固氮、产生物膜、产淀粉酶和纤维素酶等促生相关性能；最后通过培养皿和盆栽试验，分析菌株对番茄幼苗生长的影响。结果分离获得一株耐盐碱菌株，16S rRNA基因序列的比对结果显示，该菌株与芽胞杆菌属（Bacillus）模式菌株Bacillus cabrialesii strain TE3^T的同源性最高，为99.72%，将其命名为Bacillus cabrialesii zp-4245。菌株zp-4245在80 g/L NaCl、pH 9.0的条件下仍可生长；该菌株具有产铁载体、固氮、产淀粉酶、纤维素酶和生物膜等促生特性。在培养皿上，接种菌株zp-4245能够显著促进番茄幼苗在不同盐碱胁迫条件下的生长：和对照组相比，在pH 5.8的条件下，处理组番茄幼苗的侧根数、地上鲜重、地下鲜重和总鲜重分别增加了284.3%、87.3%、118.4%和94.2%；在pH 8.0的条件下，处理组番茄幼苗的侧根数、地上鲜重、地下鲜重和总鲜重分别增加了361.7%、92.3%、141.7%和101.9%；在pH 8.0+2.925 g/L NaCl的胁迫条件下，处理组番茄幼苗的侧根数增加了707.6%。盆栽番茄的促生试验表明：在非盐胁迫条件下，和对照组相比，接种菌株zp-4245处理组番茄的地上鲜重和株高分别增加了11.7%和13.9%；在6 g/kg NaCl的胁迫条件下，和对照组相比，处理组番茄的地下鲜重和株高分别增加了28.6%和10.7%。结论菌株zp-4245具有良好的耐盐碱与植物促生能力，可在不同盐碱条件下促进番茄幼苗的生长，为开发适用于盐碱地改良的微生物肥料提供了菌种资源和理论基础。

关键词: 植物根际促生细菌, 盐碱地, Bacillus cabrialesii, 番茄

Abstract:

Objective This study aimed to isolate and screen salt-alkali tolerant plant growth-promoting rhizobacteria, thereby providing microbial resources and theoretical foundations for the development of functional microbial fertilizers and the remediation of saline-alkali soils. Method Rhizosphere soil samples were collected from cotton plants in Yingjisha County, Xinjiang. Salt-alkali tolerant bacteria were isolated and screened using the serial dilution method. The strains were identified by 16S rRNA gene sequencing, and their taxonomic status was determined based on morphological, physiological, and biochemical characteristics. Salt-alkali tolerance of the strain was assessed using media with NaCl concentrations of 0–200 g/L and pH values of 4.0–13.0. Plant growth-promoting traits, such as phosphate solubilization, siderophore production, nitrogen fixation, biofilm formation, and amylase and cellulase activities, were evaluated using specific functional media. Finally, the effect of the strain on tomato seedling growth was examined through plate and pot experiments. Result We isolated a salt-alkali tolerant strain. 16S rRNA gene sequence analysis showed that the strain shared 99.72% similarity with the type strain Bacillus cabrialesii TE3^T, and we identified it as B. cabrialesii zp-4245. Strain zp-4245 could grow under conditions of 80 g/L NaCl and pH 9.0, and exhibited multiple plant growth-promoting traits, including siderophore production, nitrogen fixation, and the production of amylase, cellulase, and biofilm. In plate assays, we found that strain zp-4245 significantly promoted the growth of tomato seedlings under different salt-alkali stress conditions. Compared with the control at pH 5.8, inoculation with the strain increased lateral root number, aboveground fresh weight, underground fresh weight, and total fresh weight by 284.3%, 87.3%, 118.4%, and 94.2%, respectively. At pH 8.0, the increases were 361.7%, 92.3%, 141.7%, and 101.9%, respectively. Under combined stress of pH 8.0 and 2.925 g/L NaCl, the lateral root number increased by 707.6% compared to the control. Pot experiments demonstrated the growth-promoting effects of strain zp-4245 on tomato plants. Under non-salt-stress conditions, inoculation with strain zp-4245 increased aboveground fresh weight and plant height by 11.7% and 13.9%, respectively. Under salt stress (6 g/kg NaCl), inoculation increased underground fresh weight and plant height by 28.6% and 10.7%, respectively, compared with the control. Conclusion B. cabrialesii zp-4245 exhibits excellent salt-alkali tolerance and plant growth-promoting capabilities, significantly enhancing tomato seedling growth under varied saline-alkaline conditions. These findings provide valuable microbial resources and a theoretical foundation for developing microbial fertilizers for saline-alkali soils improvement.

Key words: plant growth-promoting rhizobacteria, saline-alkali soil, Bacillus cabrialesii, tomato

何浩迪, 梁振普, 张国只, 房世杰, 张小霞. 一株耐盐碱卡氏芽胞杆菌的分离及其促生功能鉴定[J]. 生物技术通报, 2026, 42(8): 312-322.

HE Hao-di, LIANG Zhen-pu, ZHANG Guo-zhi, FANG Shi-jie, ZHANG Xiao-xia. Isolation and Plant Growth-promoting Characterization of a Salt-alkali Tolerant Bacillus cabrialesii Strain[J]. Biotechnology Bulletin, 2026, 42(8): 312-322.

图/表 9

图1 菌株zp-4245的形态鉴定A：菌落形态；B：革兰氏染色

Fig. 1 Morphological identification of strain zp-4245A: Colonial morphology; B: gram stain

图2 基于16S rRNA基因序列构建的菌株zp-4245 的系统发育树括号内编号为菌株的 GenBank 登录号。分支上的数字为bootstrap值，代表分类单位被聚在一起的概率

Fig. 2 Phylogenetic tree of strain zp-4245 based on 16S rRNA gene sequencesGenBank accession numbers are shown in parentheses. Bootstrap values (shown at the branches) represent the probability of the taxa being clustered together

表1 菌株zp-4245生理生化测定结果

Table 1 Physiological and biochemical analysis of strain zp-4245

测定指标 Test indicators	结果 Results
葡萄糖Glucose	+
木糖Xylose	-
果糖Fructose	+
阿拉伯糖Arabinose	-
蔗糖Sucrose	+
半乳糖Galactose	-
过氧化氢酶Catalase	+
氧化酶Oxidase	-
明胶液化Gelatin liquefaction	+

图3 菌株zp-4245的盐碱耐受能力测定

Fig. 3 Salt-alkali tolerance of strain zp-4245

图4 菌株zp-4245的生长曲线

Fig. 4 Growth curve of strain zp-4245

图5 菌株zp-4245 的促生特性A：产淀粉酶能力；B：产铁载体能力；C：产纤维素酶能力；D：固氮能力。标尺=1 cm

Fig. 5 Growth-promoting characteristics of strain zp-4245A: Amylase-producing ability. B: Siderophore-producing ability. C: Cellulase-producing ability. D: Nitrogen-fixing ability. Bar=1 cm

图6 菌株zp-4245的产生物膜能力检测结果CK：未接种菌株的LB液体培养基培养3 d后的结果；zp-4245：接种菌株zp-4245的LB液体培养基培养3 d后的结果

Fig. 6 Test results of biofilm formation ability of the strain zp-4245CK: Liquid LB medium without bacterial inoculation after 3 days of culture. zp-4245: Liquid LB medium inoculated with strain zp-4245 after 3 days of culture

图7 培养皿上菌株zp-4245对番茄的促生作用A：菌株zp-4245对番茄的促生效果；B：番茄侧根数和鲜重的统计结果

Fig. 7 Growth-promoting effect of strain zp-4245 on tomato in plate assaysA: Growth-promoting effect of strain zp-4245 on tomato. B: Lateral root number and fresh weight of tomato. T1: pH 5.8, T2: pH 8.0, T3: pH 8.0+0.168 g/L NaHCO3, T4: pH 8.0+2.925 g/L NaCl, T5: pH 8.0+0.168 g/L NaHCO3+2.925 g/L NaCl. * P<0.05, **** P＜0.000 1

图8 菌株zp-4245在不同盐胁迫条件下对番茄幼苗的促生作用A：菌株zp-4245对番茄的促生效果；B：番茄的鲜重、株高和根长的统计结果。图中百分数表示各处理组与CK组相比的增长率

Fig. 8 Growth-promoting effect of strain zp-4245 on tomato seedlings under different salt stress conditionsA: Growth-promoting effect of strain zp-4245 on tomato. B: Fresh weight, plant height, and root length of tomato. The percentages in the figure indicate the growth rate of each treatment group compared to the CK group

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