Isolation of Endophytic Bacteria from Blueberry and Its Alleviative Effects on Brassica chinensis L. under Aluminum Stress

doi:10.13560/j.cnki.biotech.bull.1985.2024-0978

Abstract

Abstract:

Objective Isolation of endophytic bacteria from blueberry and investigation of their mitigating effects on aluminum stress in Brassica chinensis L. growth are aimed to provide strain resources for improving the acid and aluminum tolerance capacity of plants. Method Tissue block culture method was used to isolate the endophytic bacteria from blueberry leaf buds, and determine their aluminum tolerance characteristics by shaker culture. The dominant strains were identified by combining morphological characteristics, physiological and biochemical characteristics, 16S rDNA and gryB genome sequencing. The toxicity of the strains was verified by hemolysis test. Result Three endophytic bacteria (Y01, Y02, and Y03) were isolated from the leaf buds of blueberry, strains Y01 and Y03 showed fine tolerance to of <1 000 μmol/L aluminum concentration. Under hydroponic conditions, Brassica chinensis L. grew best at 100 μmol/L aluminum concentration, but 300 μmol/L aluminum concentration caused severe impact on B. chinensis L., and the biomass significantly reduced. Three strains of endophytic bacteria alleviated the aluminum toxicity to varying degrees, and the best effect was achieved by strain Y01, which increased the height, root length, fresh weight and chlorophyll content of B. chinensis L. by 95.6%, 40.2%, 594.7%, 22.2%. The activities of SOD, POD and CAT, and the contents of soluble sugar and soluble protein in the leaves of B. chinensis L. significantly increased by 30.8%, 7.8%, 78.3%, 91.7% and 202.5%, respectively. The activity of SOD, POD and CAT and that content of soluble sugar and soluble protein in root system increased by 118.2%, 101.2%, 59.6%, 79.2% and 74.3%, respectively,and maintained at a high level. Strain Y01 was identified as Bacillus cereus, and the hemolytic test was positive for this strain. Conclusion Endophytic bacteria Y01 isolated from blueberry alleviates the toxic effects of aluminum stress on cell membranes and other organs of B. chinensis L. by increasing chlorophyll content, antioxidant enzyme activities and release of soluble substances. This strain may improve the acid and aluminum resistance of plants as a new strain resource.

Key words: aluminum stress, endophytic bacteria, Brassica chinensis L., aluminum-resistance mechanism, cell membrane permeability, antioxidant enzyme activity

TONG De-li, ZHANG Xin, CHEN Jia-qing, HE Hai-sheng. Isolation of Endophytic Bacteria from Blueberry and Its Alleviative Effects on Brassica chinensis L. under Aluminum Stress[J]. Biotechnology Bulletin, 2025, 41(4): 302-311.

Figures/Tables 9

Fig. 1 Bacterial growth at different concentrations of aluminum ions (OD600)Y01, Y02, Y03: Three dominant bacterial strains isolated from blueberry leaf buds. Different letters indicate significant difference (P<0.05). The same below

Fig. 2 Effects of different strains on the growths of Brassica chinensis L. at five aluminium concentrationsAl0‒Al300: Aluminum ion concentration in Brassica chinensis L. hydroponic solution 0‒300 μmol/L

Fig. 3 Effects of different strains on biomass and chlorophyll contents at five aluminum concentrations

Fig. 4 Effects of different strains on conductivity at five aluminium concentrations

Fig. 5 Effects of strain Y01 on the antioxidant enzyme activities of leaves and roots of B. chinensis L.A‒C: B. chinensis L. leaves. D‒F: B. chinensis L. roots. * indicates P<0.05; ** indicates P<0.01. The same below

Fig. 6 Effects of strain Y01 on the soluble sugars and soluble proteins of leaves and roots of B. chinensis L.A, C: B. chinensis L. leaves. B, D: B. chinensis L. roots

Fig. 7 Morphological observations of the strainA, B, and C are the morphologies of Y01, Y02, and Y03 colonies; D, E, and F are the Gram staining of Y01, Y02, and Y03 under microscope

Table 1 Selected physiological and biochemical characteristics of three strains of blueberry endophytic bacteria

生理生化反应 Physiological and biochemical reaction	Y01	Y02	Y03
淀粉水解 Starch hydrolysis	+	+	+
明胶水解 Gelatin hydrolysis	+	+	+
葡萄糖水解 Glucose hydrolysis	+	-	+
接触酶活性 Contactase activity	+	+	+
V-P 反应 The V-P reaction	+	-	-
溶血试验 Hemolysis test	+	/	/

Fig. 8 Multigene phylogenetic tree of strain Y01 based on 16S rDNA sequence and gyrB gene sequence

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