Chloroplast Isolation from Fromrice Leaves

The rice chloroplast is the site of photosynthesis and the synthesis of various hormones in rice. Many nuclear-encoded proteins and chloroplast-encoded proteins play important functions in the chloroplast. The study of rice leaf color mutants can explore more genes related to chloroplast development and lay a foundation for the analysis of the leaf color regulatory network. Researchers have successfully isolated a nuclear gene associated with variegation from a naturally occurring white-spiked rice mutant, an excellent model for exploring communication between the nucleus, mitochondria, and chloroplasts in plant cells. In addition, the study found that TSCD11 encodes a seryl-tRNA synthetase located in the chloroplast, which is essential for early chloroplast development in rice under high temperature, which is helpful to further study the molecular mechanism of chloroplast development under high temperature. Often, the isolation of intact rice leaves is the first step in conducting chloroplast genetic studies. The traditional methods for extracting chloroplasts from higher plants include sucrose density gradient centrifugation, chlorinated absolute density gradient centrifugation and anhydrous method, etc., but they all have shortcomings, or the yield is low, or the purity is not high, and they are time-consuming and difficult to operate. Therefore, there is a need to develop a simple, efficient, and reproducible method to isolate intact chloroplasts from rice leaf blades.

Fig. 1. Phenotypic and genetic analysis of wp3 mutant in rice leaves. (Li H, et al., 2018)

Services

Rice is one of the important food crops for humans, and some precisely controlled mechanisms have been developed to regulate chloroplast function in its ears. Currently, little information is available on chloroplast regulation during rice panicle development. For many years, Lifeasible has been working on the independent isolation of chloroplasts from different plant tissues. We offer the isolation of chloroplasts from fresh rice leaves in a variety of media (sucrose medium, glutaraldehyde medium, sorbitol medium). In addition, our scientists are committed to developing a fast and low-cost method to obtain intact chloroplasts for the isolation and extraction of chloroplasts from C3 and C4 plants using a high-salt and low-pH method. Chloroplasts isolated by this method have been successfully used to study chloroplast-encoded proteins and mine more chloroplast development-related genes. The basic process of fromrice chloroplast isolation is as follows:

(1) Preparing fresh young leaves of fresh rice and remove the midrib of rice leaves, grind the leaves to a fine powder.

(2) Intact chloroplasts were obtained by differential centrifugation.

(3) The shape and size of chloroplasts in the precipitate were observed with a microscope.

(4) Determination of chlorophyll content.

Competitive Advantages

• The chloroplast yield can reach 89% and the purity is high.
• Quick and easy, eliminating costly and time-consuming steps such as traditional sucrose density gradient centrifugation and chlorinated blunt density gradient centrifugation.
• The requirements for the instrument are not high and the cost is low.
• It is suitable for experimental teaching and scientific research activities.
• The isolated chloroplasts had a higher nitrogen-to-chlorophyll ratio and cytoplasmic adsorption.

Lifeasible can meet the needs of customers on time and on budget through a wide range of chloroplast isolation strategies. Our aim is to be customer-centric and to provide the highest quality service to customers around the world. Our skilled and dedicated scientific researchers ensure that the most appropriate methods and techniques are selected for each specialized chloroplast project. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.

References

1. Li H, Ji G, Wang Y, et al. (2018) WHITE PANICLE3, a novel nucleus-encoded mitochondrial protein, is essential for proper development and maintenance of chloroplasts and mitochondria in rice[J]. Frontiers in plant science. 9: 762.
2. Morita K, Kono M. (1972) Isolation of chloroplasts from rice leaves using glutaraldehyde[J]. Soil Science and Plant Nutrition. 18(6): 225-231.