Will chloroplast isolation and extraction affect the structure of chloroplasts?

Generally, it will not be damaged, after stirring with a stirrer, the organelles should be separated by differential centrifugation——— which is the key centrifugation is the concentration difference, if the matrix is damaged and flows out, then there is a concentration difference! So the chloroplast is not damaged. In addition, you see that the chloroplast is large and small, that may be caused by the morphology of the chloroplast, we all know that the chloroplast is flattened, you may look at the flat side, and the vertical side is small, and the light and effect of plants depend on the receiving area of the chloroplast and sunlight to adjust the light and rate.

So what you see is a kind of accident,

Definitely, because only when the pigment is released can it be extracted and separated.

That day, I did a chloroplast separation and extraction experiment

It should be that you didn't control the concentration of the solution during extraction, and did you use salt water and sucrose for extraction? How can you do that experiment? It doesn't seem like that.

Expensive, but you can add quartz, i.e. silica, to reduce damage!

Chloroplasts can only be seen with a microscope, and what you get by slicing with your bare hands is not mesophyll cells.

Of course, it depends on your extraction technique.

The specific structure of <> chloroplasts is as follows:

1. Chloroplast is the organelle that carries out photosynthesis in the mesophyll cells of green plants, which is likened to a "nutrient manufacturing factory" and "energy conversion station";

2. Observing the chloroplasts of higher plants under an optical microscope, you can see that they are generally flattened ellipsoidal or spherical. Under the electron microscope, it can be seen that the outside of the chloroplast has a double membrane, which separates the inside of the chloroplast from the outside world, and the inside of the chloroplast contains several to dozens of basal grains;

3. The substrate is filled with substrate between the substrate and the substrate. Each substrate of a chloroplast is stacked by a cyst-like structure, and on the film of the cyst-like structure, there are pigments that carry out photosynthesis, and these pigments can absorb, transmit and convert light energy;

4. The chloroplast substrate and matrix contain many enzymes necessary for photosynthesis, and the substrate also contains a small amount of DNA.

1. Extract chlorophyll.

Chlorophyll is an ester of chlorophyllin, under the action of alkali, it can make its ester bond saponified to produce chlorophyllic acid salt, which can be soluble in water, but because it retains the structure of the mg nucleus, it still retains its original green color.

2. Isolate chlorophyll.

The water-soluble anthocyanins can be filtered out by the principle of similar miscibility, and then carotene, lutein and chlorophyll can be separated by thin layer chromatography, column chromatography and high performance liquid chromatography.

Structure of chloroplasts: chloroplasts are composed of three parts: chloroplast envelope, thylakoid, and stroma.

1. Outer coat: The chloroplast is surrounded by two layers of smooth unit membranes. The two membranes are separated by a brighter space with low electron density.

These two layers of unit membranes are called chloroplast membranes or outer envelopes. The chloroplast membrane is filled with a flowing matrix (stroma) with many lamella structures in the matrix.

2. Thlakoids: Each lamellae is composed of two layers of membranes that are closed around it, which are flat cyst-shaped, called thylakoids. Within the thylakoids is an aqueous solution.

Small thylakoids are stacked on top of each other to form a strigotryp, and such thylakoids are called sclerotoid thylakoids. The sheets that make up the basal grains are called basal lamellae. Large thylakoids traverse the stroma and run between two or more basal granules.

3. The matrix is the liquid in the space between the inner membrane and the thylakoid, and the main components include enzymes related to carbon assimilation, such as ribulose carboxylase 1,5-bisphosphate, which accounts for 60% of the total soluble protein in the matrix. In addition, there are chloroplast DNA, protein synthesis systems, and certain particle components, such as various RNA, ribosomes and other proteins.

Features:

Photosynthesis is the process by which chlorophyll absorbs light energy and converts it into chemical energy while using carbon dioxide and water to make organic matter and release oxygen. This process can be expressed by the following chemical equation: 6CO2 + 6H2O (light, enzyme, chloroplast) C6H12O6 (CH2O) + 6O2.

It includes many complex steps, and the general excavation family is divided into two stages: light reaction and dark reaction. Light reaction: This is the process by which pigment molecules such as chlorophyll absorb, transfer light energy, convert light energy into chemical energy, and form ATP and NADPH.

In this process, the water molecules are broken down and oxygen is released.

Dark reaction: The next step in photosynthesis takes place in the dark, which can also be in the light. It is a process that uses ATP formed by photoreaction to provide energy, Nadph2 reduces CO2, fixes the formed intermediate products, and makes carbohydrate roll-off products such as glucose.

1. The separation of chloroplasts should be done in isotonic solution (sodium chloride.

or sucrose. solution) to avoid osmotic pressure.

chloroplasts are damaged. The homogenate was centrifuged at 1000 r min for 2 min to remove tissue debris and some of the broken intact cells.

2. Then centrifuge at 3000 r min for 5min to obtain precipitated chloroplasts (mixed with part of the nucleus). The separation process is best carried out under the condition of 0 4; If at room temperature, quickly separate and observe.

Hello landlord. When the chromatography solution diffuses to the pigment line through capillary action, the pigment is dissolved in the chromatography solution, their solubility is not the same, when the chromatography solution continues to diffuse forward on the paper, it will continue to volatilize, so the amount of solvent will continue to decrease, so the pigment with low solubility will be precipitated first and presented on the paper, and the high solubility will be precipitated at the previous point as the chromatography solution continues to diffuse forward.

As for why there is no pigment between the two bands, it is because the chromatography fluid decreases slowly, and each pigment has only one concentration of precipitation, so only one band is formed.

You can understand it this way, chromatography liquid is like a bus, carrying 4 kinds of pigments at the starting station, each pigment can only get off at a certain station according to its own properties, so the traces will only precipitate and precipitate on the paper when it just reaches a certain pigment precipitation condition. In other places, the chromatography does carry pigment when it walks by, but the pigment is soluble at this time and does not leave traces on the paper.

Hope it helps, hope, thank you

Because the solubility of pigments in the chromatography solution is different, the diffusion rate on the filter paper strip is different, and the speed of the same type of pigment is the same, so the corresponding pigment band appears at a fixed position, rather than the pigment will appear in **. According to your understanding, the same pigment has multiple diffusion speeds, which is not scientifically reasonable.