Isolation and screening of phosphate-solubilizing bacteria from the rhizosphere of Trifolium pratense and culture medium optimization

doi:10.11686/cyxb2018064

Abstract

Abstract: In order to optimize isolation and screening of phosphate solubilizing bacteria from the rhizosphere soil of Trifolium pratense (Cv. Minshan), a preliminary extraction using Pikovaskaia’s, PKOC1 and PKOC2 phosphate solubilizing media was carried out, and the 16S rRNA gene sequence was determined for the bacterial strains with excellent phosphate solubilizing ability. The separation and screening effect of PKOC2 was better than that of PKOC1 and Pikovaskaia’s. The composition of the culture medium was optimized using a response surface experiment design. The optimal composition so identified for the PKOC2 medium was: glucose 18.19 g·L^-1, Ca₃(PO₄)₂ 5 g·L^-1, MgCl₂·6H₂O 3.21 g·L^-1, MgSO₄·7H₂O 0.25 g·L^-1, KCl 0.2 g·L^-1, (NH₄)₂SO₄ 0.08 g·L^-1. It was found that 26 strains of phosphate solubilizing bacteria were isolated from T. pratense rhizosphere using these techniques. A preliminary identification of 7 isolated bacterial strains with excellent phosphate solubilizing ability using the strains 16S rRNA gene sequence indicated that a strain designated MHS4 was Bacillus cereus, strain MHS7 and MHS19 were Bacillus subtilis, strain MHS27 was Bacillus thuringiensis, strain MHS30 was Pseudomonas fluorescens, strain MHS31 was Pseudomonas gessardii, and strain MHS49 was Klebsiella oxytoca. To summarise, phosphate-solubilizing bacteria were abundant in the rhizosphere of T. pretense were abundant, and the optimized phosphate-solubilizing medium provided support for the screening of bacterial strains with excellent phosphate-solubilizing ability.

Key words: Trifolium pratense, phosphate-solubilizing bacteria, response surface design, medium optimizied

LI Hai-yun, YAO Tuo, ZHANG Rong, ZHANG Jie, LI Zhi-yan, RONG Liang-yan, LU Xiao-wen, YANG Xiao-lei, XIA Dong-hui, LUO Hui-qin. Isolation and screening of phosphate-solubilizing bacteria from the rhizosphere of Trifolium pratense and culture medium optimization[J]. Acta Prataculturae Sinica, 2019, 28(1): 170-179.

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