Identification of a Uranium-resistant Strain and Its Growth-promoting Properties

doi:10.13560/j.cnki.biotech.bull.1985.2022-1031

Abstract

Abstract:

To provide excellent strain resources for the microbial-phytoremediation technologies of uranium-contaminated soil, and to solve the problem of uranium pollution treatment in the decommissioned mining area, the uranium-resistant and plant growth-promoting bacteria were isolated from the soil collected from the decommissioned uranium mine. Gradient dilution of contaminated soil in a decommissioned uranium mine was conducted, and a uranium-resistant strain B2 was isolated on uranium-containing nutrient agar. Then the preliminary identification of strain B2 was carried out by morphological observation, physiological, and biochemical experiments, and 16S rDNA sequence analysis. To analyze the uranium tolerance and uranium adsorption capacity of strain B2, the growth curves and uranium content of the mediums under uranium stress were measured by spectrophotometric method. The abilities of nitrogen fixation, phosphate solubilization, cellulase production, and siderophore were determined by plate methods. The production of 3-indole acetic acid（IAA）was determined by Salkowski colorimetric method, and the growth-promoting ability was verified by seed germination and potting experiment. The morphological observations, physiological and biochemical characteristics, and evolutionary analysis based on 16S rDNA sequences showed that strain B2 was a Microvirga makkahensis sp., and its growth curves conformed to the S-shaped growth curve model at a uranium concentration of 0-400 mg/L. Its growth was inhibited when the uranium concentration reached 600 mg/L. It showed no adsorption effect on uranium. Strain B2 had the probiotic properties of nitrogen fixation, phosphate solubilization, cellulase production, siderophore, and IAA production. IAA production reached 24.39 μg/mL after 48 h incubation. Strain B2 significantly improved the seed germination rate of Brassica chinensis L. and promoted the growth and development of seedlings, significantly increased the biomass of B. chinensis L. Strain B2 is a uranium-resistant M. makkahensis sp. with IAA production and other growth-promoting abilities, which can effectively promote the germination of B. chinensis L. seeds and seedling growth, and can be a candidate strain for the in plant-microbe remediation of uranium-contaminated soil.

Key words: uranium-resistant, plant growth-promoting, Microvirga makkahensis, 3-indole acetic acid（IAA）, seed germination, seedling growth, biomass, Brassica chinensis L.

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.

Figures/Tables 8

References 43

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生化特征Biochemical characteristics	B2	Microvirga makkahensis SV1470^T[26]	M. lupini Lut6^T[27]	M. aerophila DSM 21344^T[28]	M. flavescens c27j1^T[29]
菌株来源Source of strain	土壤	土壤	根瘤	空气	土壤
培养基Medium	在NA、R2A、LB生长良好	在改良的Bennett's、察氏、NA、ISP2、YMA、Rauff、R2A生长良好	在1/2LA、YMA、TY、NA生长良好	在R2A、NA生长良好，在TSA、2216E不生长	在R2A、PYE生长良好，在TSA、NA不生长
菌落颜色Color of colony	浅粉色	浅粉色	淡橙色	浅粉色	淡黄色
革兰氏染色Gram stain	G-	G-	G-	G-	G-
细胞形状Cell shape	棒状	棒状	棒状	棒状	棒状
好氧性试验 Aerobic test	严格好氧	严格好氧	严格好氧	严格好氧	严格好氧
产孢子Producing spores	-	-	-	-	-
运动性Motility	-	+	-	-	+
细胞长×细胞宽Cell length（μm）× Cell width（μm）	(1.1-1.9)×(0.5-0.7)	(1.0-1.6)×(0.8-1.3)	(1.0-2.2)×(0.4-0.5)	(1.6-4.2)×(0.8-1.1)	(1.8-4.2)×(0.4-0.7)
细胞色素氧化酶 Cytochrome oxidase	+	/	-	+	+
过氧化氢酶Oxidase	+	+	-	+	+
产吲哚Producing indoles	+	/	w	-	-
产H₂S Producing hydrogen sulphide	-	/	-	/	/
明胶液化Hydrolysis of gelatin	+	+	-	-	-
硝酸盐还原Nitrate reduction	+	-	-	-	+
柠檬酸盐利用Citrate utilization	+	/	-	/	/
脲酶Urease	-	-	+	-	-
淀粉酶Amylase	/	-	-	+	-
β-半乳糖苷酶 β-galactosidase	-	/	-	-	/
精氨酸双水介酶Arginine hydrophilic enzyme	-	+	-	-	-

生化特征Biochemical characteristics	B2	Microvirga makkahensis SV1470^T[26]	M. lupini Lut6^T[27]	M. aerophila DSM 21344^T[28]	M. flavescens c27j1^T[29]
菌株来源Source of strain	土壤	土壤	根瘤	空气	土壤
培养基Medium	在NA、R2A、LB生长良好	在改良的Bennett's、察氏、NA、ISP2、YMA、Rauff、R2A生长良好	在1/2LA、YMA、TY、NA生长良好	在R2A、NA生长良好，在TSA、2216E不生长	在R2A、PYE生长良好，在TSA、NA不生长
菌落颜色Color of colony	浅粉色	浅粉色	淡橙色	浅粉色	淡黄色
革兰氏染色Gram stain	G-	G-	G-	G-	G-
细胞形状Cell shape	棒状	棒状	棒状	棒状	棒状
好氧性试验 Aerobic test	严格好氧	严格好氧	严格好氧	严格好氧	严格好氧
产孢子Producing spores	-	-	-	-	-
运动性Motility	-	+	-	-	+
细胞长×细胞宽Cell length（μm）× Cell width（μm）	(1.1-1.9)×(0.5-0.7)	(1.0-1.6)×(0.8-1.3)	(1.0-2.2)×(0.4-0.5)	(1.6-4.2)×(0.8-1.1)	(1.8-4.2)×(0.4-0.7)
细胞色素氧化酶 Cytochrome oxidase	+	/	-	+	+
过氧化氢酶Oxidase	+	+	-	+	+
产吲哚Producing indoles	+	/	w	-	-
产H₂S Producing hydrogen sulphide	-	/	-	/	/
明胶液化Hydrolysis of gelatin	+	+	-	-	-
硝酸盐还原Nitrate reduction	+	-	-	-	+
柠檬酸盐利用Citrate utilization	+	/	-	/	/
脲酶Urease	-	-	+	-	-
淀粉酶Amylase	/	-	-	+	-
β-半乳糖苷酶 β-galactosidase	-	/	-	-	/
精氨酸双水介酶Arginine hydrophilic enzyme	-	+	-	-	-

处理Treatment	株高Plant height/cm	根长Root length/cm	茎粗Stem thickness/mm	叶片数Number of leaves	叶宽Leaf width/cm	叶长Leaf length/cm	鲜重Fresh weight		干重 Dry weight
处理Treatment	株高Plant height/cm	根长Root length/cm	茎粗Stem thickness/mm	叶片数Number of leaves	叶宽Leaf width/cm	叶长Leaf length/cm	地上Above ground/g	地下Underground/g	地上Above ground/g	地下Underground/g
CK	13.68±1.38	12.12±2.45	3.50±0.50	4.40±0.89	3.38±0.69	5.51±1.16	2.53±0.94	0.44±0.25	0.38±0.16	0.09±0.02
B2	17.35±0.97**	18.70±4.51*	5.25±0.50**	6.75±0.50**	5.23±0.59**	7.58±1.19**	9.23±2.54**	2.40±0.75**	0.86±0.12**	0.28±0.07**

处理Treatment	株高Plant height/cm	根长Root length/cm	茎粗Stem thickness/mm	叶片数Number of leaves	叶宽Leaf width/cm	叶长Leaf length/cm	鲜重Fresh weight		干重 Dry weight
处理Treatment	株高Plant height/cm	根长Root length/cm	茎粗Stem thickness/mm	叶片数Number of leaves	叶宽Leaf width/cm	叶长Leaf length/cm	地上Above ground/g	地下Underground/g	地上Above ground/g	地下Underground/g
CK	13.68±1.38	12.12±2.45	3.50±0.50	4.40±0.89	3.38±0.69	5.51±1.16	2.53±0.94	0.44±0.25	0.38±0.16	0.09±0.02
B2	17.35±0.97**	18.70±4.51*	5.25±0.50**	6.75±0.50**	5.23±0.59**	7.58±1.19**	9.23±2.54**	2.40±0.75**	0.86±0.12**	0.28±0.07**