Method for extracting chloroplast of kok-saghyz

Abstract

The invention discloses a method for extracting chloroplast from kok-saghyz leaves. The method comprises the following steps: collecting complete leaves of kok-saghyz plants; crushing the leaf tissues, and performing primary extraction to obtain a kok-saghyz chloroplast crude extract; adding the kok-saghyz chloroplast crude extract into the upper layer of the cell separation liquid, and then carrying out ultra-high-speed centrifugation for the first time at 4 DEG C for 10-15 minutes; after centrifugation is finished, sucking the precipitates of the cell separation liquid, washing the precipitates with chloroplast extracting liquid and then performing second-time ultra-high-speed centrifugation for 12 min, and obtaining the complete kok-saghyz chloroplast through the precipitates. The method can be applied to kok-saghyz chloroplast separation, can also be used for extracting chloroplast of compositae or congeneric plants, is not only suitable for being collected from common plant materials, but also can be used for chloroplast separation of recalcitrant plant leaves.

Description

technical field [0001] The invention belongs to the technical field of plant tissue identification, in particular to a method for extracting rubber grass chloroplasts. Background technique [0002] Rubber grass, where the mevalonate (MVA) pathway is used to regulate natural rubber biosynthesis, rubber grass is diploid (2n=16) with a relatively simple genome of 1.29 Gb in length containing 46,731 predicted proteins Encoding genes, rubber grass roots contain high-molecular-weight rubber molecules, and its properties are comparable to rubber latex of the Brasilian tree, which can be used as an alternative rubber crop. Rubber grass roots have a high rubber content, ranging from 3% to 28% of their total dry weight, and they are also A new source of inulin (linear β-(2-1)-linked fructans). In addition, rubber grass can grow widely in cold and temperate regions with a relatively short life cycle, and is easy to harvest and perform genetic transformation, so it can be used as an id...

AI Technical Summary

Problems solved by technology

The acquisition of intact chloroplasts is the premise of carrying out chloroplast energy conversion, electron transfer, carbon fixation and downstream omics research, but the extraction of rubber grass chloroplasts is still blank, which seriously restricts the study of the physiological and molecular mechanisms of cortical photosynthesis. Therefore, Using rubber grass as a material, the extraction and purification system of rubber grass chloroplasts will be established, which will promote the research on the photosynthesis mechanism of rubber grass at the organelle, subcellular organelle and molecular levels

Images