MOESM1 of Inoculation insensitive promoters for cell type enriched gene expression in legume roots and nodules
Creators: name: Srdjan Gavrilovic
name: Yan, Zhe
name: Jurkiewicz, Anna
name: Stougaard, Jens
name: Markmann, Katharina
Publisher: name: Figshare
Year: 2016
Subjects: subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Biotechnology
subject: Ecology
subject: Immunology
subject: FOS: Clinical medicine
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: 110309 Infectious Diseases
subject: FOS: Health sciences
subject: FOS: Health sciences
subject: Plant Biology

description: Additional file 1: Figure S1. Activity patterns of promoter candidates showing epidermis enriched (Figure S1.1), general cortex- or endodermis enriched (Figure S1.2), pericycle and phloem enriched (Figure S1.3) or xylem specific (Figure S1.4) expression in L. japonicus roots. Figure S1.4 g-l show roots expressing a control vector devoid of a promoter:GUS expression cassette. Agrobacterium rhizogenes induced transgenic roots were harvested three days after mock treatment (mock) with medium or inoculation with M. loti bacteria. Representative GUS stained root tips (a,d,g,j,m,p entire root mounts) and responsive zone fragments (b,e,h,k,n,q entire root mounts; c,f,i,l,o,r cross sections) are shown. Cross sections are 7-8 µm microtome sections of resin (Kulzer Technovit 7100) embedded roots stained with 0.1 % Ruthenium Red. Scale bars 50 µm. Figure S2. Root tip associated GUS signal varied with root hair emergence patterns in pLeExt1:GUS expressing roots. Where root hairs developed near the root tip, epidermal cells showed GUS activity in a distinct ring of blue encircling the root tip. GUS stained and fixed tips of Agrobacterium rhizogenes induced transgenic roots harvested a three days after mock treatment (mock) with medium or b inoculation with M. loti bacteria are shown. Scale bars 50 µm. Figure S3. Vibratome sections of pAtCortex:GUS expressing roots show strong enrichment of GUS signal in cortical cells. a–b 60-80 µm sections of the responsive zone of pAtCortex:GUS expressing L. japonicus roots. Roots were stained, fixed and embedded in 2.4 % agarose prior to sectioning. Scale bars 50 µm. Figure S4. pAtSUC2 activity pattern in transgenic L. japonicus roots. a–b roots shielded from light access show no GUS staining. c–d chloroplast-containing roots exposed to light show GUS signal in phloem cells. Arrows indicate GUS signal in chloroplast containing roots. Representative longitudinal views of GUS stained whole root mounts are shown. e cross-section of resin (Kulzer Technovit 7100) embedded root responsive zone stained with 0.1 % Ruthenium Red. Black arrows point to phloem poles, white arrows to pericycle cells showing GUS signal. The respective root had been exposed to light during development. Scale bars a–d 100 µm, e 50 µm. Figure S5. Promoter activity distribution across developmental zones of representative roots stably expressing promoter:GUS constructs. Transgenes are a pLeExt1:GUS (line LIG29-2); b pAtCortex:GUS (line CoG11-6); c pAtE29:GUS (line 29G4b-5); d pAtS13:GUS (line 13G26b-28) and e pAtS18:GUS (line XG5A-5). Transgenic plants were uninoculated and harvested as well as GUS stained at three weeks post germination. Scale bars 1 mm. Figure S6. Promoter activity distribution across developmental zones of representative nodulated roots stably expressing promoter:GUS constructs. Transgenes are a pLeExt1:GUS (line LIG29-2); b pAtCortex:GUS (line CoG11-6); c pAtE29:GUS (line 29G4b-5); d pAtS13:GUS (line 13G26b-28) and e pAtS18:GUS (line XG5A-5). Transgenic plants were harvested and GUS stained at two weeks after inoculation with M. loti. Scale bars 1 mm.