SupplementalMaterialsandMethods及技术讲解.docxVIP

Supplemental Materials and MethodsMicrographs of arabidopsis (Arabidopsis thaliana) chloroplastsArabidopsis (Col-0)plants were grown in growth chambers (Percivial, USA) at 21°C in a 12:12 day/night photoperiod at 125 μmol m2 s-1. Mature leaves were harvested at ED, and cut into 2 mm sections.Leaf sections were fixed and imbedded essentially as described by Pfister et al., 2014. Briefly, leaf sections were incubated in 2% (v/v) glutaraldehyde / 0.05 M sodium cacodylate buffer pH 7.4 at 4°C overnight. Sections were washed three times in with 0.1M cacodylate buffer, and dehydrated in a series of aqueous ethanol solutions from 50 to 100% (v/v) ethanol. Sections were embedded in Spurr epoxy resin (Agar Scientific), and 0.5μm thin sections were cut with a diamond knife. Sections were stained with toluidine blue for 20 min at room temperature and imaged by light microscopy.Growth conditionsand plant materialFor all experiments, the genetically broad accession, TP1245, from Agroscope (Reckenholz, Switzerland) was used. Plant material was clonally propagated by splitting 60 to 90 day old plants. To maintain growth uniformity, clones were dissected into cuttings that only contained one apical meristem and one root meristem. Clones were grown for 30 days prior to harvesting. The plant growth medium was a mix of soil, peat and perlite at a ratio of 3:2:1. Partially sunny and cloudy simulated growthconditions were carried out in a glasshouse. Direct intense sunlight was shaded to produce uniformity across the experimental growth area. The intensity of the uniform indirect sunlight was between 50 to 150 μmol m-2 s-1, depending on time of day and position of the sun. For the partially sunny growth conditions, the indirect sunlight was supplemented with 200 μmol m-2 s-1 additional light intensity from Clean Arc? metal halide lamps (EYE Lighting International, USA). Therefore,the partially sunny simulated conditions had a total intensity of 250 to 350 μmol m-2 s-1, and cloudy simula