Identification of marneral synthase, which is critical for growth and development in Arabidopsis
Article first published online: 1 OCT 2012
© 2012 The Authors. The Plant Journal © 2012 Blackwell Publishing Ltd
The Plant Journal
Volume 72, Issue 5, pages 791–804, December 2012
How to Cite
Go, Y. S., Lee, S. B., Kim, H. J., Kim, J., Park, H.-Y., Kim, J.-K., Shibata, K., Yokota, T., Ohyama, K., Muranaka, T., Arseniyadis, S. and Suh, M. C. (2012), Identification of marneral synthase, which is critical for growth and development in Arabidopsis. The Plant Journal, 72: 791–804. doi: 10.1111/j.1365-313X.2012.05120.x
- Issue published online: 22 NOV 2012
- Article first published online: 1 OCT 2012
- Accepted manuscript online: 8 AUG 2012 11:25AM EST
- Received 20 March 2012; revised 28 July 2012; accepted 6 August 2012; published online 1 October 2012.
Figure S1. Expression of genes related toflowering time in wild-type and mrn1 leaves. Total RNA wasisolated form 3-week-old wild-type and mrn1 leaves andconverted into first-strand cDNAs that were used as templates toPCR amplify transcripts of FKF1, TOC1,EMF1, ZTL, GI, CO, SOC1, FT,PHYA, PHYB, CRY1, CRY2, FLK,LD, FPA, FLC, GAI, and RGA withprimers listed in Lim et al. (2004). The elf4A(At3g13920) and ACTIN7 (At5g09810) genes wereused as a control.
Figure S2. Mapping of T-DNA insertion sites andexpression in MRN1 gene. (a) Structure of MRN1(At5g42600) and T-DNA insertion site. Boxes and linesindicate exon and intron of the MRN1 gene, respectively.Arrows indicate the sites for specific primers, which were used inRT-PCR analysis. (b) RT-PCR analysis of MRN1 gene inwild-type and mrn1 roots. Total RNA was isolated formwild-type and mrn1 roots and 100 ng of total RNA wasused in RT-PCR analysis. The ACTIN2 (At3g18780) gene was used as a control.
Figure S3. Expression of cell-cycle relatedgenes in Arabidopsis wild-type and mrn1 leaves. Total RNAwas isolated from 3-week-old wild-type and mrn1 leaves andwere converted into first-strand cDNAs that were used as templatesto PCR amplify transcripts of KRP1, KRP2, KRP3, KRP4, KRP5,KRP6, KRP7, CycB1;2, and CycD3 genes with specificprimers listed in Table S2. The Actin7 (At5g09810)gene was used as a control.
Figure S4. Expression of BR and sterolmetabolic genes in wild-type and mrn1 roots (a) and leaves(b). Total RNA was isolated from 2~3 weeks old roots and leaves inArabidopsis wild-type and mrn1 plants and converted intofirst-strand cDNAs that were used as templates to PCR amplifytranscripts of BR and sterol metabolic genes (FK, SMT2, DWF7,DWF5, DWF1, DWF6, DWF4, DWF3, BR6ox1, CYP710A1, CYP710A2) withspecific primers listed in Table S3. The ACTIN7 (At5g09810) gene was used as a control.
Figure S5. The simplified biosynthetic pathways of sterol, steroid hormone, and nonsteroidal triterpenoids in plants. Abbreviations: HMG-CoA, 3-hydroxy-3-methylglutaryl-CoA; HMGR, 3-hydroxy-3-methylglutaryl-CoA reductase; LUP1, lupeol synthase; MRN1, marneral synthase; MVA, mevalonate; SQE, squalene epoxidase; bAS1, b-amyrin synthase.
Table S1. Back-Crossing ofmrn1a Mutant with Wild-Typea Plant.
Table S2. Quantification of endogenous sterolsand triterpenols in Arabidopsis wild-type and mrn1.
Table S3. List of the oligonucleotide primers used in this study.
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