植物学技术

There are numerous options for monitoring ATP synthesis in chloroplasts using isolated thylakoid membranes, intact chloroplasts, and even whole leaves. Currently, the most commonly used method employs isolated thylakoids coupling the synthesis of ATP to light emission from luci ...

Measurement of the different components of the proton motive force (pmf) gives information about the coupling of proton movement within thylakoids to chemiosmotic processes such as photophosphorylation and protein transport, as well as that relating to the overall quality of a thyla ...

Photosynthesis is one of the most readily measured metabolic processes in a plant, with fluxes being measurable non-invasively even under field conditions. In this chapter, two principal approaches are described to measure photosynthesis – O2 evolution as determined using an O2 elect ...

Chloroplasts of plants contain an intricate membrane system, the thylakoids, which harbor the complexes of the photosynthetic machinery. Chloroplasts are confined by two membranes, the inner and outer envelope. The major glycerolipids of chloroplasts are the glycolipids monog ...

This chapter outlines a quantitative proteomics workflow using a label-free spectral counting technique. The workflow has been tested on different aspects of chloroplast biology in maize and Arabidopsis, including chloroplast mutant analysis, cell-type specific chloropl ...

Proteomics of chloroplast ribosomes in spinach and Chlamydomonas revealed unique protein composition and structures of plastid ribosomes. These studies have suggested the presence of some ribosomal proteins unique to plastid ribosomes which may be involved in plastid-unique ...

Plastoglobules are particles specifically located inside different types of plastids. They mainly contain lipids and proteins and are physically attached to thylakoids. Proteomic studies have underlined the role of plastoglobules in diverse plastid metabolic pathways, su ...

The posttranslational modification of proteins is important for the regulation of enzymatic activity, protein half-life, and interaction with other molecules. One of the best understood posttranslational modifications is the reversible phosphorylation of proteins at ser ...

Chlorophyll afluorescence has long been used as a noninvasive means to assess photosynthetic performance in plants. Pulse-amplitude modulated (PAM) fluorometry is one of the most common techniques used to study the induction and quenching of chlorophyll fluorescence in physiol ...

As part of a project to analyze chloroplast functional networks systematically, we have subjected mutants in 3,200 nuclear genes predicted to encode chloroplast-targeted proteins in Arabidopsis thaliana (http://www.plastid.msu.edu) to parallel phenotypic assays. Detail ...

Chloroplasts are the chemical factories of plant cells because they are able to fix inorganic carbon and �convert it to a wide-range of photoassimilates that are exported to the cytosol and other sub-cellular compartments. If the regulation of these processes is to be understood, the in vivo con ...

The vast majority of the several thousands of chloroplast proteins are encoded by nuclear genes. Regulation of their expression involves control of their transcription, and thus requires the transmission of information from chloroplast to nucleus (retrograde signalling). The mo ...

Proteomics aimed at addressing subcellular fractions, such as chloroplasts, are a complex challenge. In the past few years, several studies in different laboratories have identified and, more recently, quantified, thousands of proteins within whole chloroplasts or chloroplast ...

The analysis of protein–protein interactions is essential for the understanding of the molecular events in enzymatic pathways, signaling cascades, or transport processes in the chloroplast. A large variety of methods are available, which range from qualitative assays allowing f ...

Protein–protein interactions play crucial roles in the execution of many cellular functions, including those in plastids. Identifying and characterising protein–protein interactions can yield valuable information regarding the function of a protein and can also contribu ...

Since its first description in 1998 (Rigaut et al., Nat Biotech 17:1030–1032, 1999), the TAP method, for Tandem Affinity Purification, has become one of the most popular methods for the purification of in vivo protein complexes and the identification of their composition by subsequent mass spe ...

Blue native polyacrylamide gel electrophoresis (BN-PAGE) is a powerful method for separating protein complexes from biological membranes under native conditions. BN-PAGE provides much higher resolution than gel filtration or sucrose density gradient centrifugation, and ...

Photosynthetic machinery in the thylakoid membrane is prone to modifications depending on �environmental, developmental, and morphological parameters. Such plasticity in the composition of the thylakoid membrane protein complexes guarantees efficient function of the ...

Plastids are semiautonomous organelles restricted to plants and protists. These plastids are surrounded by a double membrane system, or envelope. These envelope membranes contain machineries to import nuclear-encoded proteins, and transporters for ions or metabolites, but are ...

For many studies regarding important chloroplast processes such as oxygenic photosynthesis, fractionation of the total chloroplast proteome is a necessary first step. Here, we describe a method for isolating the stromal, the thylakoid membrane, and the thylakoid lumen subchloro ...