Journal: Plant Physiology

Article Title: ANTHOCYANIDIN REDUCTASE promotes physical dormancy in Medicago truncatula seeds

doi: 10.1093/plphys/kiaf525

Figure Lengend Snippet: Isolation and characterization of anr mutant seeds in Medicago truncatula . A) RT-qPCR analysis of ANR expression in different plant organs in 3 independent experiments, error bars represent standard deviation (SD). B–E) GUS staining of seed coat at 16 DAF of WT and different stages of transgenic M. truncatula plants carrying the proANR::GUS construct, scale bar = 500 μ m. C, E) Cross-section of GUS stained seed coat, scale bar = 20 μ m. F) Schematic representation of ANR gene structure and Tnt1 insertion sites, filled black boxes represent exons and lines between them denote introns. G) RT-PCR analysis of MtANR transcript levels in WT and anr mutant, ACTIN used as the control. H) Morphologies of mature seeds from WT and anr mutants, scale bar = 500 μ m. I) Time-course analysis of seed coat development in WT and anr mutant, scale bar = 500 μ m.

Article Snippet: Based on the M. truncatula gene expression atlas ( INRAE/CNRS Medicago Expression Atlas ), the expression of ANR (ecotype A17, Mtr.44985.1.

Techniques: Isolation, Mutagenesis, Quantitative RT-PCR, Expressing, Standard Deviation, Staining, Transgenic Assay, Construct, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Plant Physiology

Article Title: ANTHOCYANIDIN REDUCTASE promotes physical dormancy in Medicago truncatula seeds

doi: 10.1093/plphys/kiaf525

Figure Lengend Snippet: Characterization of anr mutant seeds lacking physical dormancy in M. truncatula . A) Germination process of WT and anr mutant ( anr -2) seeds, with and without scarification treatment, in sterile water for 2 d. B) Weight of absorbed water of WT and anr mutant at multiple timepoints. min, minutes. C) Representative photographs of WT and anr mutant seeds during the imbibition process in sterile water, scale bar = 500 μ m. The WT seed from is the same as that shown in . D) Mature seed morphologies of WT, anr mutant, and the complementation line in the anr mutant background, scale bar = 500 μ m. E) Germination process of WT, anr mutant, and the complementation line seeds without scarification treatment. Error bars represent SD in this figure.

Article Snippet: Based on the M. truncatula gene expression atlas ( INRAE/CNRS Medicago Expression Atlas ), the expression of ANR (ecotype A17, Mtr.44985.1.

Techniques: Mutagenesis, Sterility

Journal: Plant Physiology

Article Title: ANTHOCYANIDIN REDUCTASE promotes physical dormancy in Medicago truncatula seeds

doi: 10.1093/plphys/kiaf525

Figure Lengend Snippet: Structural difference of the seed coat between the anr mutants and the WT in M. truncatula . A–H) SEM observation of mature seed coats from WT and anr mutants in M. truncatula . A, C, E, and G) Scale bar = 1 mm, B , D , F, and H : Scale bar = 2 μ m. I–L) Cross-sections of WT (I , K) and anr mutant (J , K) stained with Oil red O at 16 and 20 DAF, scale bar = 10 μ m. The arrow points to visible lipid droplets in I and K .

Article Snippet: Based on the M. truncatula gene expression atlas ( INRAE/CNRS Medicago Expression Atlas ), the expression of ANR (ecotype A17, Mtr.44985.1.

Techniques: Mutagenesis, Staining

Journal: Plant Physiology

Article Title: ANTHOCYANIDIN REDUCTASE promotes physical dormancy in Medicago truncatula seeds

doi: 10.1093/plphys/kiaf525

Figure Lengend Snippet: Content of lipid monomers in WT and anr mutant of M. truncatula . MGDG: monogalactosyldiacylglycerol; DGDG: digalactosyldiacylglycerol; SQDG: sulfoquinovosyldiacylglycerol. The vertical axis represents concentration ( μ g/g). Error bars represent SD, asterisks indicate significant differences between WT and anr mutant using 2-tailed unpaired Student's t -test (* P < 0.05; ** P < 0.01).

Article Snippet: Based on the M. truncatula gene expression atlas ( INRAE/CNRS Medicago Expression Atlas ), the expression of ANR (ecotype A17, Mtr.44985.1.

Techniques: Mutagenesis, Concentration Assay

Journal: Plant Physiology

Article Title: ANTHOCYANIDIN REDUCTASE promotes physical dormancy in Medicago truncatula seeds

doi: 10.1093/plphys/kiaf525

Figure Lengend Snippet: Different flavonoids accumulated during seed coat development in M. truncatula . A) Heatmap of differential metabolites in the flavonoid synthesis pathway compared with WT; Orange represents increase, dark orange represents new compounds in anr mutant, blue represents decrease; B) Content of the 4 highest flavonoids. Error bars represent SD, asterisks indicate significant differences between WT and anr mutant using 2-tailed unpaired Student's t -test (* P < 0.05; ** P < 0.01). DFR, dihydroflavonol 4-reductase; UGT78D2, flavonol 3- O -glucosyltransferase; BZ1, anthocyanidin 3- O -glucosyltransferase; 6''- O -acyltransferase; 3RT, anthocyanidin 3- O -rutinosyltransferase; 3GGT, anthocyanidin 3- O -glucoside 2''- O -glucosyltransferase; UGT75C1, anthocyanidin 3- O -glucoside 5- O -glucosyltransferase; UGT79B1, anthocyanidin 3- O -glucoside 2'''- O -xylosyltransferase; 3MaT1, anthocyanin 3- O -glucoside-6''- O -malonyltransferase; 5GT: cyanidin 3- O -rutinoside 5- O -glucosyltransferase; 3′GT: anthocyanin 3′- O -beta-glucosyltransferase.

Article Snippet: Based on the M. truncatula gene expression atlas ( INRAE/CNRS Medicago Expression Atlas ), the expression of ANR (ecotype A17, Mtr.44985.1.

Techniques: Mutagenesis

Journal: Plant Physiology

Article Title: ANTHOCYANIDIN REDUCTASE promotes physical dormancy in Medicago truncatula seeds

doi: 10.1093/plphys/kiaf525

Figure Lengend Snippet: Analysis of seed physical dormancy in double mutants in M. truncatula . A) Seed permeability analysis of WT, anr , tt8 , f3′5′h1 , ans , anr tt8 , anr f3′5′h1, and anr ans mutant seeds, scale bar = 500 μ m. B) Seed permeability analysis of WT and anr , tt8 , f3′5′h1 , ans , anr tt8 , anr f3′5′h1, and anr ans mutants.

Article Snippet: Based on the M. truncatula gene expression atlas ( INRAE/CNRS Medicago Expression Atlas ), the expression of ANR (ecotype A17, Mtr.44985.1.

Techniques: Permeability, Mutagenesis

Journal: Plant Physiology

Article Title: ANTHOCYANIDIN REDUCTASE promotes physical dormancy in Medicago truncatula seeds

doi: 10.1093/plphys/kiaf525

Figure Lengend Snippet: The content of lipid (A) and flavonoid (B) in WT and mutants of M. truncatula . The vertical axis represents concentration ( μ g/g). Error bars represent SD, asterisks indicate significant differences using a 2-tailed unpaired Student's t -test in this figure (* P < 0.05; ** P < 0.01).

Article Snippet: Based on the M. truncatula gene expression atlas ( INRAE/CNRS Medicago Expression Atlas ), the expression of ANR (ecotype A17, Mtr.44985.1.

Techniques: Concentration Assay