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First published online 13 August 2003
doi: 10.1242/dev.00681

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A network of redundant bHLH proteins functions in all TTG1-dependent pathways of Arabidopsis

Fan Zhang*,{dagger}, Antonio Gonzalez*, Mingzhe Zhao, C. Thomas Payne{ddagger} and Alan Lloyd§

Molecular Cell and Developmental Biology, and The Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX 78712, USA



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  		Fig. 1. SEM of seedlings at the expanded first and second leaf stage. (A) Wild type. (B) gl3-1. (C) egl3-1. (D) gl3-1 egl3-1 double mutant. (E) ttg1-1. (F) CaMV35S::GL3 cDNA in ttg1-1. (G) CaMV35S::EGL3 genomic clone in ttg1-1. (H) CaMV35S::GL3/CaMV35S::EGL3 in ttg1-1. (I) CaMV35S::GL3 cDNA in wild type. (J) CaMV35S::EGL3 genomic clone in wild type. All plants are in the Ler background.

 


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  		Fig. 2. Seed coat, seedling and flower phenotypes of mutants and transformants. (A-I) Scanning electron micrographs of seed coats illustrating the columella phenotypes. (J-L) Ruthenium red-stained seed coat mucilage phenotypes. (M-O) Seed coat pigment phenotypes. (P-R) 5-day-old seedlings. (S,T) First and second leaf stage seedlings. (U) Single flowers. (A) gl3-1 single mutant (looks like wild type). (B) gl3-1 egl3-1 double mutant. (C) egl3-1 single mutant. (D) tt8 single mutant (looks like wild type). (E) egl3-1 tt8 double mutant. (F) egl3-1 gl3-1 tt8 triple mutant. (G) ttg1-1. (H) CaMV35S::EGL3 in ttg1-1. (I) CaMV35S::GL3 in ttg1-1. (J) gl3-1 single mutant (looks like wild type); (K) gl3-1 egl3-1 double mutant (looks like egl3-1 single mutant); (L) egl3-1 tt8 double mutant. (M) Left: wild type; right: ttg1-1 mutant. (N) Lower left: CaMV35S::GL3 in ttg1-1; lower right: CaMV35S::EGL3 in ttg1-1; upper center: hybrid expressing CaMV35S::GL3 and CaMV35S::EGL3 in ttg1-1. (O) Lower center: tt8; upper left: CaMV35S::GL3 in tt8; upper right: CaMV35S::EGL3 in tt8. (P-R) Seedlings in order left to right: (P) wild type, ttg1-1, gl3-1, egl3-1, gl3-1 egl3-1 double mutant; (Q) ttg1-1, CaMV35S::GL3 in ttg1-1, CaMV35S::EGL3 in ttg1-1, hybrid expressing CaMV35S::GL3 and CaMV35S::EGL3 in ttg1-1; (R) tt8, CaMV35S::GL3 in tt8, CaMV35S::EGL3 in tt8. (S) CaMV35S::GL1 in wild type. (T) Hybrid expressing CaMV35S::GL1 and CaMV35S::EGL3 in wild type. (U) Left: PAP1-D flower (wild type expressing CaMV35S::PAP1, activation tagged); right: flower of hybrid coexpressing CaMV35S::PAP1 and CaMV35S::EGL3. Arrows in the first three panels point to examples of columellae types: C, normal round-topped columella; CC, collapsed columella; PCC, partially collapsed columella; MC, missing columella; PM, patchy mucilage; MM, missing mucilage.

 


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  		Fig. 3. SEM of root hair phenotypes. (A) Wild type. (B) gl3-1 egl3-1 double mutant. (C) ttg1-1 mutant. All are in the Ler background. Hair files are marked with *, non-hair files are marked with –. Scale bar: 50 µm.

 


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  		Fig. 4. Gene expression studies. (A-E) In situ RT-PCR of (A) EGL3 expression, (B) control minus primers, (C) GL3::GUS expression pattern, (D) GL2::GUS expression in wild type and (E) GL2::GUS expression in egl3 gl3 mutant. (F) LUX RT-PCR analysis of CHS and DFR. Gene expression levels relative to ttg1 are given above the bars. CT values and s.d. are given below the genotype and test gene labels.

 


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  		Fig. 5. Regulatory network model. This reticulated model shows all of the known bHLH and myb transcriptional regulators that function the TTG1-dependent pathways. The model illustrates the potential (broken lines) and demonstrated (unbroken lines) interactions among the proteins and genes, the narrowing of the specificity of the regulatory function as one goes from WD to bHLH to myb, and the redundancies demonstrated at the bHLH and single myb levels.

 

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© The Company of Biologists Ltd 2003