杉木林土壤和苗木内生溶磷细菌的筛选及其溶磷特性

doi:10.11707/j.1001-7488.20201201

摘要/Abstract

摘要：

目的: 筛选和鉴定杉木高效溶磷菌，研究其溶磷特性，探讨其生态适应性和溶磷特点，为研制杉木适用的微生物肥料提供菌种资源和培养条件。方法: 利用PVK无机磷培养基从杉木林地土壤和杉木植株分离溶磷菌，筛选出高效溶磷菌并结合生理生化试验和16S rDNA基因序列对其进行鉴定，采用单因素试验分析高效溶磷菌在不同pH、温度、碳源、氮源、磷源等培养条件下的溶磷特性。结果: 1）共分离筛选出20株溶磷菌，其中溶磷能力最强的3株菌株为P5、RP2和RP22（分别为195.61 mg·L^-1、109.20 mg·L^-1、78.86 mg·L^-1）。2）通过生理生化试验，结合16S rDNA序列分析及系统发育树的构建，菌株P5鉴定为Burkholderia ubonensis，RP2为Pseudomonas frederiksbergensis，RP22为Pseudomonas grimontii。3）单因素试验溶磷培养条件研究结果显示，P5菌株在pH为5，温度为30℃，碳源为蔗糖，氮源为氯化铵，磷源为磷酸钙时溶磷能力最强；RP2菌株在pH为7，温度为30℃，碳源为葡萄糖，氮源为硝酸钾，磷源为磷酸钙时溶磷能力最强；RP22菌株在pH为7，温度为25℃，碳源为葡萄糖，氮源为硝酸钾，磷源为磷酸钙时溶磷能力最强。结论: 所筛选溶磷菌具有高效溶磷能力，其中菌株P5在酸性和高温条件下仍有较高的溶磷能力，在微生物肥料研制中具有巨大潜力。

关键词: 杉木, 溶磷菌, 筛选, 鉴定, 培养条件

Abstract:

Objective: This study aimed to screen and identify efficient phosphate solubilizing bacteria from Chinese fir, We investigated its phosphate solubilizing characteristics, and explored ecological adaptability, so as to provide bacterial resources and culture conditions for the development of microbial fertilizer for Chinese fir. Method: Phosphate solubilizing bacteria were isolated from soil and Chinese fir plant by PVK inorganic phosphorus medium, and identified by physiological and biochemical tests as well as 16S rDNA gene sequence. A single factor test was used to analyze the phosphate solubilizing characteristics of phosphate solubilizing bacteria under different pH, temperature, carbon source, nitrogen source and phosphorus source. Result: 1) A total of 20 strains of phosphate solubilizing bacteria were isolated and screened, and the three strains with the strongest phosphate solubilizing capacity were P5, RP2 and RP22 (195.61, 109.20 and 78.86 mg·L^-1, respectively). 2) Strain P5 was identified as Burkholderia ubonensis, RP2 as Pseudomonas frederiksbergensis and RP22 as Pseudomonas grimontii by physiological and biochemical tests, combined with 16S rDNA sequence analysis and phylogenetic tree construction. 3) The results of single factor experiment showed that the P5 strain had the strongest phosphate solubilizing ability when the pH was 5, the temperature was 30℃, the carbon source was sucrose, the nitrogen source was ammonium chloride, and the phosphorus source was calcium phosphate. RP2 showed the strongest phosphate solubilizing ability when pH7, 30℃, the carbon source was glucose, the nitrogen source was potassium nitrate, and the phosphorus source was calcium phosphate. RP22 showed the strongest phosphate solubilizing ability when the pH7, 25℃, the carbon source was glucose, the nitrogen source was potassium nitrate, and the phosphorus source was calcium phosphate. Conclusion: The screened bacteria have high phosphate solubilizing ability among which strain P5 has an efficient phosphate solubilizing ability under acidic and high temperature conditions, which has great potential for the development of microbial fertilizers.

Key words: Chinese fir, phosphate solubilizing bacteria, screening, identification, culture conditions

中图分类号:

S714.3

韦宜慧,陈嘉琪,赵光宇,董玉红,厚凌宇,焦如珍. 杉木林土壤和苗木内生溶磷细菌的筛选及其溶磷特性[J]. 林业科学, 2020, 56(12): 1-9.

Yihui Wei,Jiaqi Chen,Guangyu Zhao,Yuhong Dong,Lingyu Hou,Ruzhen Jiao. Screening of Phosphate Solubilizing Bacteria from Soil and Endogenous Environment of Chinese Fir Seedlings and Their Characterization of Phosphate Solubilization[J]. Scientia Silvae Sinicae, 2020, 56(12): 1-9.

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菌株 Strain No.	分离部位 Isolated part	形状 Shape	颜色 Color	表面状态 Surface state	菌株 Strain No.	分离部位 Isolated part	形状 Shape	颜色 Color	表面状态 Surface state
P1	S	C	T	M	P11	S	C	W	M
P2	S	C	W	M	P12	S	C	W	M
P3	S	C	T	M	P13	S	C	T	M
P4	S	C	T	M	SP13	St	C	LY	M
P5	S	C	W	M	LP2	L	C	W	D
P6	S	C	T	M	RP2	R	C	W	D
P7	S	C	W	D	RP13	R	C	W	M
P8	S	C	W	M	RP22	R	C	LY	M
P9	S	C	W	M	RP23	R	C	PY	SM
P10	S	C	PY	M	RP24	R	C	W	M

菌株 Strain No.	可溶性磷含量 Soluble phosphorus content	菌株 Strain No.	可溶性磷含量 Soluble phosphorus content
P1	21.61±1.82h	P11	50.77±2.45e
P2	52.68±4.86e	P12	47.08±1.29f
P3	45.67±3.94f	P13	51.73±1.97e
P4	39.62±3.59f	SP13	65.17±5.52d
P5	195.61±14.9a	LP2	44.29±2.09f
P6	56.63±4.29e	RP2	109.20±3.06b
P7	49.54±1.10e	RP13	68.22±5.26d
P8	46.54±3.00f	RP22	78.86±5.02c
P9	36.63±2.85g	RP23	74.54±6.68c
P10	24.13±1.81h	RP24	67.83±4.97d

菌株 Strain No.	革兰氏染色 Gram staining	葡萄糖水解 Glucose hydrolysis	乳糖水解 Lactose hydrolysis	甲基红 Methyl red	V-P试验 V-P test	硫化氢试验 Hydrogen sulfide production	明胶液化 Gelatin liquefaction	柠檬酸盐 Citrate	丙二酸盐 Malonate	反硝化 Denitrification
P1	-	+	+	+	-	-	+	+	-	-
P2	-	+	+	-	-	-	-	-	-	+
P3	-	+	+	+	-	-	+	+	-	+
P4	-	+	+	+	-	-	+	+	-	+
P5	-	+	+	-	-	-	-	+	-	+
P6	-	+	+	+	-	-	+	+	-	+
P7	+	+	+	+	-	-	+	+	-	-
P8	-	+	+	+	-	-	+	+	-	+
P9	-	+	+	+	-	-	+	+	-	+
P10	-	+	+	+	-	-	+	+	-	-
P11	-	+	+	+	-	-	+	+	-	-
P12	-	+	+	+	-	-	+	+	-	+
P13	-	+	+	+	-	-	+	+	-	+
SP13	-	+	+	+	-	-	+	+	-	-
LP2	-	+	+	-	+	-	+	+	-	-
RP2	-	+	+	-	+	-	+	+	-	+
RP13	-	+	+	+	-	-	+	+	-	+
RP22	-	+	+	-	+	-	+	+	+	+
RP23	-	+	-	-	-	+	-	-	-	-
RP24	-	+	-	+	-	-	-	-	-	-

菌株 Strain No.	模式菌株 Type strain	相似性 Identity(%)
P1	副伯克氏菌 Paraburkholderia hiiakae (JF763857^T)	98.78
P2	伯克氏菌 Burkholderia humptydooensis (MSMB43^T)	96.52
P3	副伯克氏菌 Paraburkholderia terrae (NBRC00964^T)	98.64
P4	伯克氏菌 Burkholderia arboris (AM47630^T)	99.92
P5	伯克氏菌 Burkholderia ubonensis (CIP107078^T)	98.44
P6	欧文氏菌 Erwinia rhapontici (ATCC29283^T)	99.11
P7	芽孢杆菌 Bacillus aryabhattai (B8W22^T)	100
P8	副伯克氏菌 Paraburkholderia metrosideri (DFBP6-1^T)	97.7
P9	伯克氏菌 Burkholderia oklahomensis (C6786^T)	98.67
P10	泛菌属 Pantoea conspicua (LMG24534^T)	99.63
P11	欧文氏菌 Erwinia billingiae (CIP106121^T)	98.81
P12	副伯克氏菌 Paraburkholderia insulsa (KF733462^T)	99.25
P13	副伯克氏菌 Paraburkholderia metrosideri (DFBP6-1^T)	98.62
SP13	泛菌属 Pantoea roadsii (LMG26273^T)	98.84
LP2	假单胞菌 Pseudomonas donghuensis (HYS^T)	98.75
RP2	假单胞菌 Pseudomonas frederiksbergensis (JAJ28^T)	99.61
RP13	副伯克氏菌 Paraburkholderia caffeinilytica (CF1^T)	99.31
RP22	假单胞菌 Pseudomonas grimontii (CFML97-514^T)	100
RP23	新鞘氨醇菌 Novosphingobium barchaimii (LL02^T)	99.69
RP24	苍白杆菌 Ochrobactrum pseudogrignonense (CCUG30717^T)	100

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