Planta

, Volume 231, Issue 6, pp 1385–1400 | Cite as

Features of anthocyanin biosynthesis in pap1-D and wild-type Arabidopsis thaliana plants grown in different light intensity and culture media conditions

Original Article

Abstract

The number of different anthocyanin molecules potentially produced by Arabidopsis thaliana and which anthocyanin molecule is the first product of anthocyanidin modification remain unknown. To accelerate the understanding of these questions, we investigated anthocyanin biosynthesis in rosette leaves of both pap1-D and wild-type (WT) A. thaliana plants grown in nine growth conditions, which were composed of three light intensities (low light, middle light, and high light) and three media derived from MS medium (medium-1, 2, and 3). These nine growth conditions differentially affected the levels of anthocyanins and pigmentation patterns of rosette leaves, which were closely related to the diversification levels of cyanin structures. The combined growth conditions of high light and either medium-2 or medium-1 induced the most molecular diversity of anthocyanin structures in rosette leaves of pap1-D plants. Twenty cyanin molecules, including five that were previously unknown, were characterized by HPLC-ESI-MS and HPLC-TOF-MS analyses. We detected that the A. thaliana anthocyanin molecule A11 was most likely the first cyanin derived from the multiple modification steps of cyanidin. In addition, in the same growth condition, rosette leaves of pap1-D plants produced much higher levels and more diverse molecular profiling of cyanins than those of WT plants. The transcript levels of PAP1, PAL1, CHS, DFR, and ANS cDNAs were much higher in pap1-D rosette leaves than in WT ones. Furthermore, on the same agar-solidified medium, an enhancement of light intensity increased levels and molecular diversity of cyanins in both pap1-D and WT rosette leaves. In the same light intensity condition, the responses of anthocyanin levels and profiling to medium alternation were different between pap1-D and WT plants.

Abbreviations

pap1-D

Production of anthocyanin pigmentation1-Dominant

PAP1

Production of anthocyanin pigmentation1

HPLC-ESI-MS

High performance-electrospray ionization-mass spectrometry

HPLC-TOF-MS

High performance-time of flight-mass spectrometry

RT-PCR

Reverse transcription-polymerase chain reaction

Keywords

Anthocyanins Anthocyanidins Arabidopsis thaliana Biosynthesis Cyanins Cyanidins HPLC-ESI-MS HPLC-TOF–MS pap1-D mutant PAP1 Metabolic profiling RT-PCR 

Supplementary material

425_2010_1142_MOESM1_ESM.jpg (941 kb)
S-Fig. 1 Normalized values from semi-quantitative analysis of RT-PCR products show expression levels of 5 genes in rosette leaves of plants grown in different growth conditions. a: Normalized values show the impacts of three light intensities on transcript levels of five genes in rosette leaves of both WT and pap1-D plants grown on agar-solidified medium-2. Due to poor growth of WT plants in the condition of HLM-2, total RNA isolated from plants grown in the condition of HLM-3 were used as controls. HLM-2 (WT:M-3) means that total RNA for pap1-D plants was isolated from leaves on the condition of HLM-2 and total RNA for WT plants was isolated from leaves on the condition of HLM-3. b: Normalized values show the impacts of three media on transcript levels of 5 genes in rosette leaves of both WT and pap1-D plants grown in the ML condition. RT-PCR analysis for WT seedlings grown on medium-3 in the ML condition is not shown due to low yield and poor quality of total RNA from rosette leaves. LLM-2: low light: 50 μmol m−2 s−1 of light intensity and medium-2: ½ strength of NH4NO3 and KNO3 in MS medium MLM-2: middle light: 200 μmol m−2 s−1 of light intensity and medium-2 HLM-2: high light: 500 μmol m−2 s−1 of light intensity and medium-2 HLM-3: high light and medium-3: 0 NH4NO3 and ½ strength of KNO3 in MS medium MLM-1: middle light and medium-1: full strength of basal MS medium MLM-3: middle light and medium-3 (JPEG 942 kb)
425_2010_1142_MOESM2_ESM.jpg (2.2 mb)
S-Fig. 2 Mass spectra of 20 cyanins detected at 530 nm from rosette leaves of 20-day-old pap1-D plants grown in the condition of HLM-2 (high light: 500 μmol m−2 s−1 of light intensity and medium-2: ½ strength of both NH4NO3 and KNO3 in MS medium) (JPEG 2248 kb)
425_2010_1142_MOESM3_ESM.doc (26 kb)
Supplementary method (DOC 26 kb)

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Department of Plant BiologyNorth Carolina State UniversityRaleighUSA

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