Correlation between spatial (3D) structure of pea and bean thylakoid membranes and arrangement of chlorophyll-protein complexes.
BACKGROUND
The thylakoid system in plant chloroplasts is organized into two distinct domains: grana arranged in stacks of appressed membranes and non-appressed membranes consisting of stroma thylakoids and margins of granal stacks. It is argued that the reason for the development of appressed membranes in plants is that their photosynthetic apparatus need to cope with and survive ever-changing environmental conditions. It is not known however, why different plant species have different arrangements of grana within their chloroplasts. It is important to elucidate whether a different arrangement and distribution of appressed and non-appressed thylakoids in chloroplasts are linked with different qualitative and/or quantitative organization of chlorophyll-protein (CP) complexes in the thylakoid membranes and whether this arrangement influences the photosynthetic efficiency.


RESULTS
Our results from TEM and in situ CLSM strongly indicate the existence of different arrangements of pea and bean thylakoid membranes. In pea, larger appressed thylakoids are regularly arranged within chloroplasts as uniformly distributed red fluorescent bodies, while irregular appressed thylakoid membranes within bean chloroplasts correspond to smaller and less distinguished fluorescent areas in CLSM images. 3D models of pea chloroplasts show a distinct spatial separation of stacked thylakoids from stromal spaces whereas spatial division of stroma and thylakoid areas in bean chloroplasts are more complex. Structural differences influenced the PSII photochemistry, however without significant changes in photosynthetic efficiency. Qualitative and quantitative analysis of chlorophyll-protein complexes as well as spectroscopic investigations indicated a similar proportion between PSI and PSII core complexes in pea and bean thylakoids, but higher abundance of LHCII antenna in pea ones. Furthermore, distinct differences in size and arrangements of LHCII-PSII and LHCI-PSI supercomplexes between species are suggested.


CONCLUSIONS
Based on proteomic and spectroscopic investigations we postulate that the differences in the chloroplast structure between the analyzed species are a consequence of quantitative proportions between the individual CP complexes and its arrangement inside membranes. Such a structure of membranes induced the formation of large stacked domains in pea, or smaller heterogeneous regions in bean thylakoids. Presented 3D models of chloroplasts showed that stacked areas are noticeably irregular with variable thickness, merging with each other and not always parallel to each other.
Department of Plant Anatomy and Cytology, Institute of Plant Experimental Biology and Biotechnology, Faculty of Biology, University of Warsaw, Miecznikowa 1, Warsaw, PL-02-096, Poland.
Izabela
I
Rumak
Radosław
R
Mazur
Katarzyna
K
Gieczewska
Joanna
J
Kozioł-Lipińska
Borys
B
Kierdaszuk
Wojtek P
WP
Michalski
Brian J
BJ
Shiell
Jan Henk
JH
Venema
Wim J
WJ
Vredenberg
Agnieszka
A
Mostowska
Maciej
M
Garstka
D002734:Chlorophyll / Q000378:metabolism; D000077194:Chlorophyll A; D061146:Chlorophyll Binding Proteins / Q000378:metabolism*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007700:Kinetics; D045342:Light-Harvesting Protein Complexes / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D058503:Mesophyll Cells / Q000166:cytology / Q000648:ultrastructure; D018613:Microscopy, Confocal; D018532:Pisum sativum / Q000378:metabolism* / Q000648:ultrastructure*; D027805:Phaseolus / Q000378:metabolism* / Q000648:ultrastructure*; D045331:Photosystem I Protein Complex / Q000378:metabolism; D045332:Photosystem II Protein Complex / Q000378:metabolism; D018515:Plant Leaves / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism; D011489:Protein Denaturation; D013050:Spectrometry, Fluorescence; D013696:Temperature; D020524:Thylakoids / Q000378:metabolism / Q000648:ultrastructure*
D061146:Chlorophyll Binding Proteins; D045342:Light-Harvesting Protein Complexes; D008565:Membrane Proteins; D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D010940:Plant Proteins; D002734:Chlorophyll; D000077194:Chlorophyll A