Isolation of Chloroplasts from Plant Tissue

Not all higher plants are suitable for the isolation and preparation of chloroplasts, the most commonly used plant materials are spinach, peas, corn, sugar beets, barley, etc. In addition, lower plants such as chlamydomonas and chlorella are also commonly used materials. Lifeasible's expertise in chloroplast engineering allows us to isolate chloroplasts from different specimens, different tissues, and even different plants (both higher and lower), and prepare the type of chloroplast preparation according to your needs. If you have any special requirements about our isolation of chloroplasts from plant tissue services, please feel free to contact us. We are looking forward to working together with your attractive projects.

Isolated Chloroplasts from Various Plants

For green plants, chloroplasts are the place for photosynthesis, so what cells do chloroplasts mainly exist in? Studies have found that chloroplasts mainly exist in the mesophyll cells of plants. The distribution of chloroplasts is relatively extensive, in addition to the mesophyll of higher plants, it also exists in the cells of some young stems, and algae cells also contain chloroplasts. If the plants are not the same, then the shape and size of the corresponding chloroplasts are also different. Isolated chloroplasts are suitable tools for studying chloroplast function. However, it is quite difficult to isolate active, relatively pure intact chloroplasts. The cells of some plant leaf cells are thicker, and the chloroplasts are not easy to separate from the cells. Some plant leaf cells contain some endogenous inhibitory substances, which are easily released during the preparation process and destroy the structure and activity of chloroplasts. Therefore, not all higher plants are suitable for the isolation and preparation of chloroplasts, and it is necessary to select suitable plant materials for chloroplast isolation. In addition, the chloroplast viability will continue to decline with the extension of the in vitro time. Therefore, the separation work should be completed in the shortest time possible.

Fig. 1. Isolating intact chloroplasts in 30% Percoll. (Islam M, et al., 2019)

Services

Lifeasible has developed detailed protocols for the isolation of many types of plant chloroplasts. After the leaves are ground and homogenized, they are separated by filtration and differential centrifugation according to their particle size. The entire separation process is carried out at 0-4°C, and all extracts, solutions and materials are also stored at this temperature for future use. At the same time, chloroplasts are prepared in isotonic solution, thereby reducing osmotic pressure damage to chloroplasts. Our chloroplast isolation services are well-received by customers around the world for high yield, purity and integrity, and small quantities of starting material. In addition, we can provide you with different types of chloroplasts according to your needs, including complete chloroplasts, unbroken chloroplasts, broken chloroplasts, free-lamellar chloroplasts, chloroplast fragmentation and sub-chloroplast particles.

With our services, you rely on the analysis of chloroplasts purified in intact form from different species of plants and tissues to elucidate many chloroplast physiological parameters, such as the properties of enzymes or biosynthetic pathways. Our scientists have successfully built various types of plant chloroplast isolation platforms to independently isolate chloroplasts from different plant tissues, including but not limited to:

• Chloroplast Isolation from Arabidopsis Thaliana Plants
• Chloroplast Isolation from Spinach Leaves
• Chloroplast Isolation from Chlamydomonas Reinhardtii
• Chloroplast Isolation from Tomato Leaves
• Chloroplast Isolation from Fromrice Leaves
• Chloroplast Isolation from Marine Microalgae

Optimized Chloroplast Isolation Methods

• Based on high salt buffer, saline Percoll gradient method.
• Based on high salt buffer, sucrose density gradient method.
• Based on high salt buffer, nitrogen-sucrose gradient method.
• Based on high sorbitol concentration buffer, Percoll gradient method.

Reference

1. Islam M, Buttelmann G L, Chekhovskiy K, et al. (2019) Isolation of intact chloroplast for sequencing plastid genomes of five Festuca species[J]. Plants. 8(12): 606.