Chloroplast Isolation from Tomato Leaves

Tomato is the most planted horticultural crop in the world, with more than 4.7 million hectares planted annually. Tomato is a model system for studying fruit development, and recent insights into tomato plastid proteome dynamics during chloroplast-to-chromosome differentiation have provided important insights into these processes. Furthermore, tomato is a model system to study the induction of plant defenses associated with injury, herbivore and pathogen attack. As we all know, chloroplasts are important organelles that sense and transmit metabolic and stress signals to different cellular components, while maintaining the location of photosynthesis and metabolic factories. Scientists have now assessed the proteome of intact leaves, chloroplasts and plastid subcellular fractions in the model plant Arabidopsis thaliana. But much less is known about the tomato chloroplast proteome or its subgroups. High-yielding tomato chloroplast and matrix isolation methods are required before proteomics-based strategies can be developed.

Fig. 1. Isolation of tomato chloroplasts on Percoll gradients. (Bhattacharya O, et al., 2020)

Services

Tomato is a widely produced crop, and bioactive compounds found in the tomato plant have been used commercially in pharmaceuticals, biopesticides, spices, and food additives, including flavorings and colorants. For many years, Lifeasible has been working on the independent isolation of chloroplasts from different plant tissues. We successfully established a tomato model system for proteomic analysis of the chloroplast matrix proteome. Our scientists are committed to developing a high-yield method to isolate intact chloroplasts from tomato leaves. The isolation protocol we adopted was based on methods developed in Arabidopsis chloroplasts and combined many of the recommendations from basic spinach and pea studies to be successful. Chloroplasts isolated by this method have been successfully used for proteomic analysis and identification of chloroplast colocalized proteins. The basic process of tomato chloroplast isolation is as follows:

(1) Tomato culture: using only undamaged young leaves from 4 to 5 week old tomato plants.

(2) Veined tomato leaf homogenate was centrifuged and the pellet contained intact and broken chloroplasts.

(3) The pellet was carefully resuspended and overlaid on a 40%-80% Percoll-Ficoll-PEG step gradient.

(4) Microscopic observation of intact chloroplasts at the Percoll interface.

(5) Chloroplast quality assessment.

Competitive Advantages

• A simplified, efficient method for chloroplast isolation.
• Suitable for three simultaneous 60 g chloroplast preparations.
• For proteomics, protein import and biochemistry, and chloroplast genome isolation studies.
• Obtaining intact, high-quality chloroplasts.
• Chloroplast membrane and stromal fractions can be recovered.

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.

Reference

1. Bhattacharya O, Ortiz I, Walling L L. (2020) Methodology: an optimized, high-yield tomato leaf chloroplast isolation and stroma extraction protocol for proteomics analyses and identification of chloroplast co-localizing proteins[J]. Plant methods. 16(1): 1-21.