Separating and screening method for siderophilic phosphorite-dissolving bacterium

A technology for separation and screening of phosphorus-solubilizing bacteria, applied in the direction of bacteria, etc., can solve the problems of difficult plant absorption and utilization, low screening efficiency, and low utilization rate of phosphate fertilizers, and achieve the goal of simplifying the separation and screening steps, shortening the screening time, and improving the separation and screening efficiency Effect

Inactive Publication Date: 2014-12-03
SHANDONG INST OF POMOLOGY
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Problems solved by technology

[0003] Phosphorus is one of the indispensable nutrient elements for plant growth. Although a large amount of soluble phosphorus fertilizer is applied to the soil every year, the phosphate in the soil is often mainly tricalcium phosphate. This kind of phosphate has poor solubility and is difficult to be absorbed by plants. , the utilization rate of phosphorus fertilizer by crops in the current season is generally only 5% to 10%, and the aftereffect of crops is added, and the utilization rate does not exceed 25%.
Separation and screening of microbial strains with high-efficiency phosphorus-dissolving effect and making micro-fertilizers is an effective method to solve the low utilization rate of phosphorus fertilizers and control the environmental pollution caused by the abuse of fertilizers
[0004] For the screening of plant growth-promoting rhizobacteria (Plant Growth-Promoting Rhizobacteria, referred to as PGPR) with iron-loving or / and phosphate-solubilizing ability, and used in agricultural production to solve the problems of plant iron deficiency yellowing and low utilization rate of phosphate fertilizer, there have been some related reports
However, in the existing technology, siderophilic bacteria and phosphate-dissolving bacteria are screened separately. Even if some strains have multiple disease prevention and growth promotion mechanisms such as siderophilia and phosphate-dissolving ability, their siderophilic ability or phosphate-dissolving ability are detected separately, and the steps are complicated. , the screening efficiency is low, time-consuming and labor-intensive

Method used

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  • Separating and screening method for siderophilic phosphorite-dissolving bacterium
  • Separating and screening method for siderophilic phosphorite-dissolving bacterium
  • Separating and screening method for siderophilic phosphorite-dissolving bacterium

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Embodiment 1

[0028] A method for separating and screening iron-loving phospholytic bacteria, comprising the following steps:

[0029] (1) Collect crop rhizosphere soil, remove impurities, inoculate in sterilized water with an inoculum size of 1.0g / 100mL, cultivate at 30°C for 2.5 hours, let stand, separate supernatant, and set aside;

[0030] (2) To prepare the CAS-P detection plate, first prepare 1mmol / L CaCl 2 solution, 1mmol / L MgSO 4 ·7H 2 O solution, acid hydrolyzed casein solution with a mass concentration of 10% (121°C, sterilized separately for 15 minutes); then take 0.2mL of CaCl 2 solution, 0.2 mL of MgSO 4 ·7H 2 O solution, 6 mL of 10% acid hydrolyzed casein solution, to which Ca 3 (PO 4 ) 2 0.5g, use biological buffer 1,4-piperazine diethanesulfonic acid [piperazine-N,N-bis(2-ethanesulfonic acid), Pipes], adjust pH 6.8~7.0. Dilute to 100mL with deionized water. Add 2g of agar powder and sterilize at 121°C for 15 minutes. After sterilization, when the temperature drops ...

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Abstract

The invention discloses a separating and screening method for a siderophilic phosphorite-dissolving bacterium. The method comprises the following steps: (1) collecting rhizosphere soil of crops, removing impurities, inoculating sterile water with the rhizosphere soil, culturing for 2-3 hours at 25-35 DEG C and separating supernate; (2) preparing a CAS-P detection flat panel, uniformly mixing a CaCl2 solution, a MgSO4.7H2O solution, and an acid hydrolyzed casein solution with the mass concentration of 8-12%, adding Ca3(PO4)2 into the mixture, adjusting the pH to 6.8-7.0 and fixing the volume by means of water; adding agar powder, sterilizing, and further adding CAS blue detection liquid when the temperature is reduced to 50-60 DEG C after sterilization, uniformly mixing and pouring on the flat panel; and (3) inoculating the CAS-P detection flat panel with the supernate obtained in the step (1), wherein the inoculum size is 100-200 microliter/flat panel, culturing for 3-5 days at 25-35 DEG C, wherein the strain growing on the detection flat panel is the siderophilic phosphorite-dissolving bacterium. By adopting the screening method disclosed by the invention, the separating and screening steps are simplified, the screening time is greatly shortened, and the separating and screening efficiency of the siderophilic phosphorite-dissolving bacterium is improved.

Description

technical field [0001] The invention relates to a method for separating and screening iron-loving phospholytic bacteria. Background technique [0002] Iron is one of the trace elements necessary for most organisms. Although the content of iron elements in the earth's crust occupies the fourth place, most of them cannot be directly used by plants, and a set of efficient iron transport mechanisms are needed to obtain the iron ions they need. In a low-iron environment, most bacteria can synthesize and secrete siderophiles for use by themselves and plants to promote plant growth; and by competing with pathogenic microorganisms for limited iron nutrition, they can inhibit the growth and reproduction of pathogenic microorganisms and play a role in biological control role. [0003] Phosphorus is one of the indispensable nutrient elements for plant growth. Although a large amount of soluble phosphorus fertilizer is applied to the soil every year, the phosphate in the soil is often ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20
Inventor 余贤美艾呈祥付丽安淼王海荣刘嘉芬
Owner SHANDONG INST OF POMOLOGY
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