Method for separating inner-layer and outer-layer substances of wetland plant root surface iron film

Abstract

The invention discloses a method for separating inner-layer and outer-layer substances of a wetland plant root surface iron film. The method comprises the following steps: S1, taking roots, shaking off soil, and washing the roots with sterile water; S2, cutting the roots into root segments and placing in a centrifuge tube, adding a PBS-S phosphate buffer solution, carrying out vortexes, then taking the roots out of the centrifuge tube, centrifuging the solution and then carrying out microbial DNA extraction of an outer layer of the root surface iron film, wherein solid substances in a suspension in the centrifuge tube are substances of the outer layer of the root surface iron film; and S3, putting the roots after the vortexes into another new centrifuge tube, adding 0.5 Mhydroxylamine-0. 5M hydrogen chloride solution and sterile glass beads into the centrifuge tube, and manually shaking the centrifuge tube up and down for 5 minutes till that the surfaces of the roots become white so asto obtain a solid substance in a suspension in the centrifuge tube, wherein the solid substance is a root surface iron film inner-layer substance. In the invention, a circulation process of inner andouter layers of substances and a change condition of microorganisms can be analyzed so that the influence of an oxygen concentration gradient on rhizosphere iron-sulfur circulation can be better revealed.

Description

technical field [0001] The invention relates to the technical field of material separation, in particular to a method for separating the inner and outer layers of iron film on the root surface of wetland plants. Background technique [0002] Post-mining mining wastelands usually contain large amounts of iron sulfides (such as pyrite FeS 2 ), its oxidative acidification will lead to acid wastewater generation, soil acidification, heavy metal release and other problems. At the same time, due to the insufficient supply of nutrients such as nitrogen, phosphorus, and potassium, it is difficult for ordinary plants to colonize. Wetland plants can transport oxygen in the atmosphere to the root surface through the root aerenchyma, so that the rhizosphere is in a relatively oxidized state, and a micro-oxidative environment is formed in the plant rhizosphere, so that the reducing substance Fe existing in the soil under flooding conditions 2+ Oxidation occurs to form a root surface iro...

AI Technical Summary

Problems solved by technology

[0004] Disadvantages of the existing technology: the existing technical scheme can only extract all the iron film on the root surface at one time, and cannot distinguish the inner and outer layer of the iron film on the root surface, and it takes a long time, the extraction is not thorough, there are many iron film residues, and the extraction efficiency is low. low

Images