Pseudomonas fluorescens Strain CLW17: Degradation of Glyphosate and Its Mechanism

doi:10.11924/j.issn.1000-6850.casb2022-1170

Abstract

Abstract:

The subject is to study the degradation of glyphosate by the rhizobacteria-promoting strain Pseudomonas fluorescens CLW17 and the molecular mechanism of the strain. The glyphosate degradation ability of strain CLW17 was evaluated by plate culture method and molybdenum antimony resistance colorimetric method. An optimized condition of degrading glyphosate by strain CLW17 was achieved by employing Plackett-Burman (PB) design, and central composite design (CCD). Bioinformatics analysis was carried out for understanding the key gene thiO in glyphosate degradation. Deletion mutant strain (CLW17ΔthiO) and complement strain (ΔthiO/thiO) were constructed, and their effects on glyphosate degradation were studied. The maximum concentration of glyphosate tolerance of strain CLW17 was 0.7%. The degradation rate of CLW17, CLW17ΔthiO and ΔthiO/thiO was 72.81%, 32.78% and 54.33%, respectively, under the optimal condition for glyphosate degradation. Bioinformatics analysis revealed that thiO gene encoded 366 amino acids and included signal peptide and transmembrane domain. CLW17 was capable of degrading glyphosate effectively. Moreover, thiO gene was closely related to glyphosate degradation ability. These findings can provide valuable bacterial resources for glyphosate pollution control and soil remediation.

Key words: Pseudomonas fluorescens, glyphosate, biodegradation, optimization, thiO gene

CLC Number:

CAO Yongqing, LIU Yan, ZHANG Lihui, JIN Tingting, REN Jiahong. Pseudomonas fluorescens Strain CLW17: Degradation of Glyphosate and Its Mechanism[J]. Chinese Agricultural Science Bulletin, 2022, 38(30): 108-117.

Figures/Tables 13

References 28

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Primers	Sequences (5’→3’)
CLW17_04770FW (Hind Ⅲ)	CCCAAGCTTCTGAGTCCAGTGGCAACATCAC
CLW17_Del04770FW	AGGAAGAAAAACTGACTAGATGGTCATGTTTGATCCAACCGT
CLW17_Del04770RV	ACGGTTGGATCAAACATGACCATCTAGTCAGTTTTTCTTCCT
CLW17_04770RV (EcoR I)	CCGGAATTCACGTGATCGAACTGCGTGTG
CLW17_04770SQFW	ACTACGCGGTGCTCGACACTAACGCTGAT
CLW17_04770SQRV	CCAACAGGAAACCGTGGTCGATGTG
CLW17_Comp04770FW -EcoR Ⅰ	CCGGAATTCACGGTTGGATCAAACATG
CLW17_Comp04770RV -Spe I	CGGACTAGTCTAGATCCGTCCGGCCGGTG
pBAD-Forward	ATGCCATAGCATTTTTATCC
T7	TAATACGACTCACTATAGGG

Primers	Sequences (5’→3’)
CLW17_04770FW (Hind Ⅲ)	CCCAAGCTTCTGAGTCCAGTGGCAACATCAC
CLW17_Del04770FW	AGGAAGAAAAACTGACTAGATGGTCATGTTTGATCCAACCGT
CLW17_Del04770RV	ACGGTTGGATCAAACATGACCATCTAGTCAGTTTTTCTTCCT
CLW17_04770RV (EcoR I)	CCGGAATTCACGTGATCGAACTGCGTGTG
CLW17_04770SQFW	ACTACGCGGTGCTCGACACTAACGCTGAT
CLW17_04770SQRV	CCAACAGGAAACCGTGGTCGATGTG
CLW17_Comp04770FW -EcoR Ⅰ	CCGGAATTCACGGTTGGATCAAACATG
CLW17_Comp04770RV -Spe I	CGGACTAGTCTAGATCCGTCCGGCCGGTG
pBAD-Forward	ATGCCATAGCATTTTTATCC
T7	TAATACGACTCACTATAGGG

因素	回归系数	效果	平方和	贡献值
intercept	34.04
X₁	9.25	18.49	1025.78	23.09
X₂	-2.29	-4.58	62.92	1.42
X₃	6.19	12.38	459.52	10.34
X₄	0.74	1.48	6.54	0.15
X₅	-11.01	-22.01	1453.54	32.72
X₆	-8.69	-17.38	906.4	20.40
X₇	2.32	4.65	64.75	1.46
X₈	4.37	8.74	229.32	5.16
X₉	-1.28	-2.57	19.76	0.44
X₁₀	2.87	5.75	99.15	2.23
X₁₁	3.10	6.19	115.05	2.59

因素	回归系数	效果	平方和	贡献值
intercept	34.04
X₁	9.25	18.49	1025.78	23.09
X₂	-2.29	-4.58	62.92	1.42
X₃	6.19	12.38	459.52	10.34
X₄	0.74	1.48	6.54	0.15
X₅	-11.01	-22.01	1453.54	32.72
X₆	-8.69	-17.38	906.4	20.40
X₇	2.32	4.65	64.75	1.46
X₈	4.37	8.74	229.32	5.16
X₉	-1.28	-2.57	19.76	0.44
X₁₀	2.87	5.75	99.15	2.23
X₁₁	3.10	6.19	115.05	2.59

实验组数	编码/预测值			实际值	预测值
实验组数	X₁/℃	X₅/(g/L)	X₆/(g/L)	实际值	预测值
1	-1/26	-1/8	-1/0.3	61.2962	59.81
2	1/30	-1/8	-1/0.3	64.2350	64.62
3	-1/26	1/12	-1/0.3	64.0738	64.43
4	1/30	1/12	-1/0.3	67.1785	65.49
5	-1/26	-1/8	1/0.7	67.0071	67.93
6	1/30	-1/8	1/0.7	66.0919	64.97
7	-1/26	1/12	1/0.7	68.8709	67.73
8	1/30	1/12	1/0.7	60.3094	61.03
9	-2/24.64	0/10	0/0.5	66.3589	66.80
10	2/31.36	0/10	0/0.5	64.5662	65.21
11	0/28	-2/6.64	0/0.5	63.2265	63.63
12	0/28	2/13.36	0/0.5	63.5127	64.18
13	0/28	0/10	-2/0.16	62.4065	63.49
14	0/28	0/10	2/0.84	66.5652	66.56
15	0/28	0/10	0/0.5	73.4263	72.78
16	0/28	0/10	0/0.5	74.3185	72.78
17	0/28	0/10	0/0.5	70.0747	72.78
18	0/28	0/10	0/0.5	73.5353	72.78
19	0/28	0/10	0/0.5	72.5685	72.78
20	0/28	0/10	0/0.5	72.9370	72.78