Journal: Molecular Plant Pathology

Article Title: Use of enhancer trapping to identify pathogen‐induced regulatory events spatially restricted to plant–microbe interaction sites

doi: 10.1111/mpp.12287

Figure Lengend Snippet: Screening of an enhancer‐trap population reveals three predominant pathogen infection‐related histochemical staining patterns. (A) An X‐gluc (5‐bromo‐4‐chloro‐3‐indolyl‐β‐d‐glucuronic acid) halo‐like pattern (blue) surrounds putative infection sites. (B) An HpaNoco2 sporangiophore emerges from a myob1‐2 X‐gluc‐stained (blue) stoma (white arrow). (C) Hyphae‐associated X‐gluc staining (blue) is seen in enhancer‐trap plant leaves infected with the virulent isolate HpaNoco2. (D) Hyaloperonospora arabidopsidis (Hpa)‐infected myob1‐2 mesophyll tissue co‐stained with trypan blue and MAGENTA‐GlcA (van Damme et al., 2008), illustrating that responses of plant cells (magenta) are restricted to H pa structures (dark blue). H pa hyphae (white arrow) and haustoria (yellow asterisks) are stained with trypan blue and β‐glucuronidase (GUS)‐expressing plant cells are stained with MAGENTA‐GlcA (black arrows).

Article Snippet: Histochemical GUS staining was performed by soaking tissues for 18 h in a 5‐bromo‐4‐chloro‐3‐indolyl‐β‐ d ‐glucuronic acid‐containing [X‐gluc (1 mg/mL; Gold Biotechnology, Inc., St. Louis, MO, USA) or MAGENTA‐GlcA (0.5 mg/mL; Duchefa Biochemie, Haarlem, the Netherlands)] solution (50 m m Na 2 PO 4 , pH 7.2, 0.5 m m K 4 Fe(CN) 6 , 0.5 m m K 3 Fe(CN) 6 , 0.05% Triton‐X) at 37 °C and cleared with 95% ethanol.

Techniques: Infection, Staining, Expressing

Journal: Molecular Plant Pathology

Article Title: Use of enhancer trapping to identify pathogen‐induced regulatory events spatially restricted to plant–microbe interaction sites

doi: 10.1111/mpp.12287

Figure Lengend Snippet: A MyoB1 (Myosin Binding Protein 1) promoter‐controlled reporter gene shows a response to oomycete infection and wounding, yet canonical plant defence pathway marker genes are not influenced directly by MyoB1. (A) MyoB1 promoter:GUS fusion transgenic lines mimic the staining patterns seen in the original enhancer‐trap line (myob1‐2). Two independent pMyoB1:GUS lines (lines 6 and 12) and myob1‐2 are shown at 7 days post‐infection (dpi) with the virulent isolate HpaNoco2 (I), at 48 h post‐wounding (II) and untreated (III). Post‐treatment, plants were placed in a solution containing X‐gluc (5‐bromo‐4‐chloro‐3‐indolyl‐β‐d‐glucuronic acid), followed by the removal of chlorophyll with ethanol. (B) PR1 and PDF1.2A are not constitutively transcribed in myob1 mutant plants. A single representative semi‐quantitative reverse transcription‐polymerase chain reaction (RT‐PCR) experiment shows transcripts from plants heavily infected with HpaNoco2 at 7 dpi (+) and from water‐treated (−) plants.

Article Snippet: Histochemical GUS staining was performed by soaking tissues for 18 h in a 5‐bromo‐4‐chloro‐3‐indolyl‐β‐ d ‐glucuronic acid‐containing [X‐gluc (1 mg/mL; Gold Biotechnology, Inc., St. Louis, MO, USA) or MAGENTA‐GlcA (0.5 mg/mL; Duchefa Biochemie, Haarlem, the Netherlands)] solution (50 m m Na 2 PO 4 , pH 7.2, 0.5 m m K 4 Fe(CN) 6 , 0.5 m m K 3 Fe(CN) 6 , 0.05% Triton‐X) at 37 °C and cleared with 95% ethanol.

Techniques: Binding Assay, Infection, Marker, Transgenic Assay, Staining, Mutagenesis, Reverse Transcription Polymerase Chain Reaction