Unlock instant, AI-driven research and patent intelligence for your innovation.

Salix matsudana branch cortex chloroplast preparation method

A chloroplast and switchgear technology, applied in the field of agricultural biology, can solve the problems such as no preparation method, public report and the like of chloroplast in the bark of Salix chinensis.

Inactive Publication Date: 2017-11-24
INST OF FORESTRY CHINESE ACAD OF FORESTRY
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is no public report on the preparation method of the chloroplasts of the willow bark

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Salix matsudana branch cortex chloroplast preparation method
  • Salix matsudana branch cortex chloroplast preparation method
  • Salix matsudana branch cortex chloroplast preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1 prepares willow branch cortex chloroplast with the method of the present invention

[0022] 1) Sampling and homogenization: in the early morning of the growing season, take the bark of the current year's branches of willow, wash it with deionized water, dry it with filter paper, and cut the bark into 5mm 2 Take 20g of bark pieces and transfer them to a homogenizer (WARING, USA), add 200mL of separation liquid (Table 1), and homogenize twice at slow speed and fast speed, each time for 5 seconds, with an interval of 1 minute.

[0023] 2) Filtration: filter the homogenate through 4 layers of gauze, and collect the filtrate in an ice-bathed beaker.

[0024] 3) Preparation of coarse chloroplast precipitate: Divide the filtrate evenly into 50 mL centrifuge tubes, centrifuge at 200 g for 3 min in an angle rotor, transfer the supernatant to a new centrifuge tube, and centrifuge at 1500 g for 10 min to obtain coarse chloroplast precipitate.

[0025] 4) Suspension o...

Embodiment 2

[0031] Example 2 Chloroplast Fluorescence Microscopic Examination

[0032] The chloroplast suspension obtained in the present invention was dropped on the center of the glass slide, covered with a cover glass to prepare a temporary mount, and examined with a fluorescence microscope (Imager.A1, Zeiss). Observe the images of chloroplasts in bright field to judge whether there is contamination of other organelles in the field of view; observe the images of chlorophyll fluorescence of chloroplasts under excitation light to judge the activity of chloroplasts. The results showed that the chloroplasts were scattered and distributed in the bright field, and no contamination of other cell components was found; under the excitation light, the chloroplasts all had chlorophyll fluorescence, and the chloroplasts had photosynthetic activity.

Embodiment 3

[0033] Embodiment 3 Chloroplast capsule integrity inspection

[0034] The potassium ferricyanide method is used to test the capsule integrity of the chloroplast obtained in the present invention. Take two equal-volume chloroplast suspensions, one of which is burst in deionized water, and the other is not subjected to bursting treatment, and the oxygen release rate of the two chloroplasts is detected with an oxygen electrode (Chlorolab2, Hansatech) respectively, and calculated according to formula 1 The integrity rate of the chloroplast capsule, it can be determined that the integrity rate of the chloroplast prepared by the method of the present invention is 87%.

[0035] Formula 1:

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of agricultural biology, and discloses a Salix matsudana branch cortex chloroplast preparation method, which comprises: 1) bark collection; 2) mechanical homogenization; 3) filtration with a gauze; 4) chloroplast crude precipitate preparation; 5) chloroplast crude precipitate suspension; 6) continuous density gradient liquid preparation; 7) iso-density centrifugation; and 8) complete chloroplast recovery. According to the present invention, the purity and the integrity of the cortex chloroplast obtained by using the method of the present invention can meet the downstream experiment requirements; and with the preparation method, the foundation is established for the research of the physiological and molecular mechanisms of Salix matsudana branch cortex photosynthesis at the organelle and molecule level, and the reference is provided for the extraction of the chloroplasts of other non-leaf photosynthetic organs.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and relates to a method for extracting and purifying the complete chloroplast of the cortex green tissue of willow willow branches. Background technique [0002] Chloroplasts in woody plant branches release CO from mitochondrial respiration and transpiration 2 Recovery and fixation, because the green photosynthetic cells are mainly located in the cortical tissue, the chloroplasts in the branches are called cortical chloroplasts, and the photosynthesis of the branches is called cortical photosynthesis. Cortical photosynthesis can release CO from branches to the atmosphere 2 Converted to carbohydrates, making plants more economical in carbon utilization. Cortical photosynthesis can reduce the damage of acidic substances in branches to basic cell structures, reduce the production of toxic substances in anaerobic respiration, and improve the survival competitiveness of trees under adversit...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N1/28
CPCG01N1/28
Inventor 刘俊祥郎蓬蓬孙振元古琳樊磊罗栋翟飞飞
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY