Table 1.
The primers used in this paper.
Fig 1.
Tissue-specific expression of PsMPT in germinated buds (A) and transcriptional levels of PsMPT in response of GA3 and ABA of tree peony by qPCR (B).
100 μmol L-1 ABA and 50 μmol L-1 GA3 was sprayed to the dormant buds in green house (18–22°C, 8-h-light/16-h-dark cycle). Values are means ±SD of three replicates.
Fig 2.
PsMPT promoter sequence of the 5’ region upstream from the start codon (ATG) and putative cis-elements predicted in the promoter region.
Numbers indicate the positions relative to the translation start codon starting from the adenosine (+1). The putative TATA box at position -189, CAAT box at position -210, and the ATG start codon is framed in box and denoted with blue color. The other important putative cis-elements are framed and labeled below.
Fig 3.
Histochemical localization (I) and tissue-specific expression of GUS (II) in transgenic Arabidopsis thaliana carrying the PsMPT promoter::GUS construct.
(I)Histochemical localization of GUS expression by staining with X-gluc in transgenic Arabidopsis thaliana carrying the PsMPT promoter::GUS construct. Arrow bar shows GUS staining in flower. A 7-day-plants from seeding; B 21-day-plants from seeding; C 28-day-plants from seeding; D Positive control (Ca35S promoter driven); E roots; F: stem; G leaf; H: mature silique; I flower; J stigma; K stamen; L petal. (II)Total RNA was isolated from roots (R), rosette leaf (RL), cauline leaf (CL), flower bud (F), bloomed flower (BF), stamen (S), stigma (St), petal (P) and siliques (Si) of 35-day-old transgenic plants from seeding. The transcriptional levels were analyzed by qPCR using GUS gene-specific PCR primers, which were normalized with beta-actin. Values are means ± SD of three replicates.
Fig 4.
Relative expression levels of PsMPT promoter in response to hormone and various abiotic stresses treatments in transgenic Arabidopsis plants.
(A) GUS expressions when exposed to 4°C temperature. Plants were transferred to a cold chamber maintained at 4°C, and the control grew at 18°C. Error bars represent ±SD. (B) GUS staining of transgenic Arabidopsis grew at 18°C (left) and at 4°C for 6 h (right). (C) GUS expressions were measured by qPCR using 35-day-old plants from seeding. 50 μmol L-1 GA3, 100 μmol L-1 NAA, 100 μmol L-1 ABA, 250 mmol L-1 ACC, 200 mmol L-1 NaCl, 40 μmol L-1 mannitol and 100 mmol L-1 PEG was sprayed to the inflorescence at 18°C, and double-distilled water treatment was used as control. (D) GUS fluorescence (nmol L-1 min-1 μg protein-1) were measured by a Microplate Spectrofluorometer using 35-day-old plants from seeding. 50 μmol L-1 GA3, 100 μmol L-1 NAA, 100 μmol L-1 ABA, 250 mmol L-1 ACC, 200 mmol L-1 NaCl, 40 μmol L-1 mannitol and 100 mmol L-1 PEG was sprayed to the inflorescence at 18°C, and double-distilled water treatment was used as control.
Fig 5.
Assays for GUS expression driven by the series PsMPT promoter.
(A) Schematic diagram of the PsMPT promoter deletions and mutation that were used to analyze the activity of different fragments of the PsMPT promoter. All fragmented promoter were fused to a GUS reporter gene. (B) Quantitative analyses of GUS expression in transgenic plants driven by deletion or muted constructs of PsMPT promoter in response to chilling. (C) GUS activity in transgenic Arabidopsis plants. The inflorescence of 5-week-old Arabidopsis plants was used as material, and five independent lines for every treatment.