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Solubilization method for isolation of photosynthetic mega- And super-complexes from conifer thylakoids
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysiologisk botanik. Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Umeå Plant Science Centre (UPSC).ORCID-id: 0000-0003-3858-4606
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.ORCID-id: 0000-0001-9831-1533
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysiologisk botanik. Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Umeå Plant Science Centre (UPSC).ORCID-id: 0000-0002-7906-6891
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.ORCID-id: 0000-0002-3691-2699
2021 (Engelska)Ingår i: Bio-protocol, E-ISSN 2331-8325, Vol. 11, nr 17, artikel-id e4144Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Photosynthesis is the main process by which sunlight is harvested and converted into chemical energy and has been a focal point of fundamental research in plant biology for decades. In higher plants, the process takes place in the thylakoid membranes where the two photosystems (PSI and PSII) are located. In the past few decades, the evolution of biophysical and biochemical techniques allowed detailed studies of the thylakoid organization and the interaction between protein complexes and cofactors. These studies have mainly focused on model plants, such as Arabidopsis, pea, spinach, and tobacco, which are grown in climate chambers even though significant differences between indoor and outdoor growth conditions are present. In this manuscript, we present a new mild-solubilization procedure for use with “fragile” samples such as thylakoids from conifers growing outdoors. Here, the solubilization protocol is optimized with two detergents in two species, namely Norway spruce (Picea abies) and Scots pine (Pinus sylvestris). We have optimized the isolation and characterization of PSI and PSII multimeric mega- and super-complexes in a close-to-native condition by Blue-Native gel electrophoresis. Eventually, our protocol will not only help in the characterization of photosynthetic complexes from conifers but also in understanding winter adaptation.

Ort, förlag, år, upplaga, sidor
2021. Vol. 11, nr 17, artikel-id e4144
Nyckelord [en]
2nd Dimension SDS-PAGE, Blue-Native gel electrophoresis, Norway spruce, Photosystem I, Photosystem II, Picea abies, Pinus sylvestris, Scots pine, Thylakoids
Nationell ämneskategori
Botanik Biokemi Molekylärbiologi
Identifikatorer
URN: urn:nbn:se:umu:diva-191367DOI: 10.21769/BioProtoc.4144ISI: 000694707900012PubMedID: 34604449Scopus ID: 2-s2.0-85116025854OAI: oai:DiVA.org:umu-191367DiVA, id: diva2:1627667
Tillgänglig från: 2022-01-14 Skapad: 2022-01-14 Senast uppdaterad: 2025-02-20Bibliografiskt granskad
Ingår i avhandling
1. How could Christmas trees remain evergreen?: photosynthetic acclimation of Scots pine and Norway spruce needles during winter
Öppna denna publikation i ny flik eller fönster >>How could Christmas trees remain evergreen?: photosynthetic acclimation of Scots pine and Norway spruce needles during winter
2022 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Alternativ titel[sv]
Hur kan julgranen vara grön? : fotosyntesapparatens anpassning till vinterförhållanden hos tall och gran
Abstract [en]

Plants and other green organisms harvest sunlight by green chlorophyll pigments and covertit to chemical energy (sugars) and oxygen in a process called photosynthesis providing the foundation for life on Earth. Although it is unanimously believed that oceanic phytoplanktons are the main contributors to the global photosynthesis, the contribution of coniferous boreal forests distributed across vast regions of the northern hemisphere cannot be undermined. Hence boreal forests account signifificantly for social, economical and environmental sustainability. Not only do conifers thrive in the tundra regions with extreme climate, but they also maintain their needles green over the boreal winter. A question remains; what makes them so resilient? In this respect, we aimed to understand the remarkable winter adaptation strategies in two dominant boreal coniferous species,i.e., Pinus sylvestris and Picea abies. First, we mapped the transcriptional landscape in Norway spruce (Picea abies) needles over the annual cycle. Transcriptional changes in the nascent needles reflflected a sequence of developmental processes and active vegetative growth during early summer and summer. Later after maturation, transcriptome reflflected activated defense against biotic factors and acclimationin response to abiotic environmental cues such as freezing temperatures during winter. Secondly, by monitoring the photosynthetic performance of Scot pine needles, we found that the trees face extreme stress during the early spring (Feb-Mar) when sub-zero temperatures are accompanied by high solar radiation. At this time, drastic changes occur in the thylakoid membranes of the chloroplast that allows the mixing of photosystem I and photosystem II that typically remain laterally segregated. This triggers direct energy transfer from PSII to PSI and thus protects PSII from damage. Furthermore, we found that this loss of lateral segregation may be a consequence of triple phosphorylationof Lhcb1 (Light harvesting complex1 of photosystem II). The structural changes in thylakoid membranes also lead to changes inthe thylakoid macro domain organisationand pigment protein composition. Furthermore, we discovered that while PSII is protected by direct energy transfer, the protection of PSI is provided through photoreduction of oxygen by flavodiiron proteins, which in turn allows P700 to stay in an oxidised state necessary for direct energy transfer. These coordinated cascades of changes concomitantly protect both PSI and PSII to maintain the needles green over the winter.

Ort, förlag, år, upplaga, sidor
Umeå: Umeå University, 2022. s. 65
Nyckelord
Scots Pine, Norway spruce, Photosynthesis, Winter adaptation
Nationell ämneskategori
Biokemi Molekylärbiologi Bioinformatik och beräkningsbiologi Botanik Biofysik Evolutionsbiologi
Forskningsämne
biologisk kemi; biologi; molekylärbiologi; fysiologisk botanik
Identifikatorer
urn:nbn:se:umu:diva-194032 (URN)978-91-7855-788-2 (ISBN)978-91-7855-787-5 (ISBN)
Disputation
2022-05-20, Lilla hörsalen, KBC huset, Umeå, 09:30 (Engelska)
Opponent
Handledare
Anmärkning

On page 12, List of papers, manuscripts (in preparation): Athor names hidden due to copyright - see printed version. 

Tillgänglig från: 2022-04-29 Skapad: 2022-04-22 Senast uppdaterad: 2025-02-20Bibliografiskt granskad

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Bag, PushanSchröder, Wolfgang P.Jansson, StefanFarci, Domenica

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