吡咯伯克霍尔德菌WY6-5的溶磷、抑菌与 促玉米生长作用研究

doi:10.3864/j.issn.0578-1752.2019.09.009

Abstract

Abstract:

【Objective】 The study was carried out to screen microbe with phosphate solubilizing (P-solubilizing) and antagonistic activity, to evaluate their efficacies for P-solubilizing, fungal inhibition and plant growth promoting, and to identify antifungal compounds, so as to provide new resources for the development of microbial fertilizers. 【Method】 Rhizosphere soil samples of tea trees were collected from Cheyun mountain factory in Xinyang, Henan, China. Each sample was diluted, and spread onto the surface of insoluble organic and inorganic media. The diameter of P-solubilizing zone was measured after 5 days of incubation. The strain WY6-5 was chosen for further studies because it showed the highest P-solubilizing activity on insoluble phosphate. Additionally, the strain WY6-5 was inoculated in liquid medium and soil for 20 days to test P-solubilizing activity, and inoculated in maize grown soil to test plant growth promoting effects. Moreover, strain WY6-5 was co-cultured with eight different fungi to determine antagonistic activity by using a two dish face-to-face cultural method. The volatiles were characterized and identified with gas chromatography-tandem mass spectrometry (GC-MS/MS). 【Result】 Three strains with P-solubilizing activity were isolated from rhizosphere soils of tea trees, and which were capable of dissolving insoluble organic and inorganic phosphorus medium. The strain WY6-5 demonstrated the highest P-solubilizing activity with the solubilizing halo up to 2.3 cm on insoluble organic medium, and 3.6 cm on insoluble and inorganic phosphorus medium. The ratios of the P-solubilizing halo diameter to the colony diameter on both media were 4.6 and 7.2, respectively. Based on morphological characteristics, physiological, biochemical and phylogenetic analyses, the strain WY6-5 was identified to be Burkholderia pyrrocinia. P-solubilizing activity of WY6-5 was also observed in the liquid medium or in soil after 20 days of incubation. The concentration of soluble phosphate in liquid medium was up to 520.4 mg·L ^-1, which was 176 times higher than that in control treatment. During 3-20 d, the phosphate concentrations in soil under WY6-5 treatments were constantly higher than that under control treatment. In addition, the strain WY6-5 significantly promoted the growth of maize seedling in terms of the number, length, width and area of leaf as well as plant height and fresh weight. Moreover, the volatile compounds produced from the WY6-5 inhibited the growth of all eight different fungi, with the mycelium inhibition rate up to 100%. The antifungal volatile was subsequently identified as dimethyl disulfide through GC-MS/MS. 【Conclusion】 The Burkholderia pyrrocinia strain WY6-5 isolated from rhizosphere soils of a tea tree was found to be able to dissolve insoluble phosphate in both liquid medium and soil, to promote the growth of maize seedlings, and to produce volatile dimethyl disulfide with broad antifungal activity, implying an important biological functions.

Key words: phosphate solubilizing, volatile antifungal activity, Burkholderia pyrrocinia, dimethyl disulfide, microbial fertilizer, maize

GONG AnDong, ZHU ZiYu, LU YaNan, WAN HaiYan, WU NanNan, Cheelo Dimuna, GONG ShuangJun, WEN ShuTing, HOU Xiao. Functional Analysis of Burkholderia pyrrocinia WY6-5 on Phosphate Solubilizing, Antifungal and Growth-Promoting Activity of Maize[J].Scientia Agricultura Sinica, 2019, 52(9): 1574-1586.

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试验内容 Experiments	WY6-5	B. cepacia / pyrrocinia*	试验内容 Experiments	WY6-5	B. cepacia/ pyrrocinia*
碳/氮源利用 Carbon /Nitrogen utilization			盐耐受性 Salt tolerance
α-D-葡糖Alpha-D-glucose	++	+	1% NaCl	++	++
D-山梨醇 D-sorbitol	+	+	4% NaCl	-	-
D-半乳糖醛酸 D-Galacturonic Acid	++	++	8% NaCl	-	-
吐温40 Tween 40	+	+	pH耐受性 pH tolerance
糊精 Dextrin	+	-	pH 5	+	++
D-果糖 D-Fructose	++	++	pH 6	++	++
L-丙氨酸L-Alanine	+	++	抗生素敏感性 Antibiotics sensitivity
乙酸 Acetic Acid	+	++	万古霉素 Vancomycin	++	++
奎宁酸Quinic Acid	++	++	林肯霉素 Lincomycin	++	++
D-天冬氨酸 D-Aspartic Acid	++	++	利福霉素SV Rifamycin SV	++	++
L-丝氨酸 L-Serine	+	+	二甲胺四环素 Minocycline	+	++
D-甘露醇 D-Mannitol	++	+	化学敏感性测试Chemical sensitivity
L-苹果酸 L-Malic Acid	++	++	盐酸胍 Guanidine HCl	+	+
L-焦谷氨酸 L-Pyroglutamic Acid	+	++	四唑蓝 Tetrazolium blue	++	++
D-葡糖酸 D-Gluconic Acid	++	++	四唑紫 Tetrazolium purple	++	++
D-阿拉伯醇 D-Arabitol	+	+	硫酸四癸钠 Niaproof 4	+	++
L-丝氨酸 L-Serine	+	+	1%乳酸钠 1% Sodium Lactate	++	++

生长指标Growth index	对照组Control group	处理组WY6-5
叶长Leaf length (cm)	26.56±0.70	32.31*±0.95
叶宽Leaf width (cm)	2.41±0.08	2.92*±0.11
单叶叶面积Area of single leaf (cm²)	48.29±2.61	71.37*±4.45
单株叶片数Number of leaves per plant	8.33±0.14	9.00*±0.17
茎粗Stem diameter (cm)	0.58±0.02	0.72*±0.03
株高Plant height (cm)	30.20±0.45	34.12*±1.21
单株鲜重Fresh weigh of single plant (g)	5.73±0.02	7.07*±0.09
地上部干重 Dry weight of overground tissues (g/plant)	1.22±0.03	1.83*±0.08
地下部干重Dry weight of underground root (g/plant)	0.31±0.02	0.36±0.03

Functional Analysis of Burkholderia pyrrocinia WY6-5 on Phosphate Solubilizing, Antifungal and Growth-Promoting Activity of Maize

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