Developing a Dwarf Echinacea purpurea Line by Suppressing DWF3 Expression via RNA Interference

Jin Zhao; Yuan-Yuan Fu; Jong-Hwa Park; Geun-Won Choi; Kong Young Park; Youn Hyung Lee; In Sik Chung

doi:10.12972/kjhst.20170067

All Issue

2017 Vol.35, Issue 5 Preview Page Next Page

Developing a Dwarf Echinacea purpurea Line by Suppressing DWF3 Expression via RNA Interference

31 October 2017. pp. 628-636

PDF XML

Abstract

DWARF3 (DWF3) is a brassinosteroid (BR) biosynthesis-related enzyme that catalyzes the C-3 oxidation of early BR intermediates. For the first time, we isolated the EpDWF3 gene from Echinacea purpurea. EpDWF3 shares high homology with AtCYP90A1 and ZeCPD1, a steroid 23-alpha-hydroxylase involved in the BR-biosynthetic pathway. We generated transgenic Echinacea plants via transformation with an RNA interference (RNAi) construct that expresses hairpin double-stranded RNA for EpDWF3 to induce sequence-specific RNA silencing. The transgenic plants displayed dwarf phenotypes, including reduced plant height and small flower diameter. We confirmed the presence of the RNAi construct in the transgenic plants by genomic DNA PCR. The transcript level of EpDWF3 decreased by 48% in the transgenic plants compared to wild type. Our results indicate that the knock-down of BR biosynthesis-related genes by RNAi enables the production of valuable dwarf plants for commercial horticultural or agricultural applications.

Keywords

brassinosteroid

knock-down

RNA silencing

steroid 23-alpha-hydroxylase

transgenic plant

References

Chung HY, Fujioka S, Choe S, Lee S, Lee YH, Baek NI, Chung IS (2010) Simultaneous suppression of three genes related to brassinosteroid (BR) biosynthesis altered campesterol and BR contents, and led to a dwarf phenotype in Arabidopsis thaliana . Plant Cell Rep 29:397-402. doi:10.1007/s00299-010-0830-z

Clouse SD, Langford M, McMorris TC (1996) A brassinosteroid insensitive mutant in Arabidopsis thaliana exhibits multiple defects in growth and development. Plant Physiol 111:671-678. doi:10.1104/pp.111.3.671

Fujioka S, Yokota T (2003) Biosynthesis and metabolism of brassinosteroids. Annu Rev Plant Biol 54:137-164. doi:10.1146/annurev.arplant.54.031902.134921

Gudesblat GE, Russinova E (2011) Plants grow on brassinosteroids. Curr Opin Plant Biol 14:530-537. doi:10.1016/j.pbi.2011.05.004

Guo H, Li L, Aluru M, Aluru S, Yin Y (2013) Mechanisms and networks for brassinosteroid regulated gene expression. Curr Opin Plant Biol 16:545-553. doi:10.1016/j.pbi.2013.08.002

Khush GS (1999) Green revolution: preparing for the 21st century. Genome 42:646-655. doi:10.1139/g99-044

Li J, Nagpal P, Vitart V, McMorris TC, Chory J (1996) A role for brassinosteroids in light-dependent development of Arabidopsis . Science 272:398-401. doi:10.1126/science.272.5260.398

Ohnishi T, Godza B, Watanabe B, Fujioka S, Hategan L, Ide K, Shibata K, Yokota T, Szekeres M, Mizutani M (2012) CYP90A1/CPD, a brassinosteroid biosynthetic cytochrome P450 of Arabidopsis , catalyzes C-3 oxidation. J Biol Chem 287:31551-31560. doi:10.1074/jbc.M112.392720

Sakamoto T, Matsuoka M (2004) Generating high-yielding varieties by genetic manipulation of plant architecture. Curr Opin Biotechnol 15:144-147. doi:10.1016/j.copbio.2004.02.003

Szekeres M, Ne´meth K, Koncz-Ka´lma´n Z, Mathur J, Kauschmann A, Altmann T, Re´dei GP, Nagy F, Schell J, Koncz C (1996) Brassinosteroids rescue the deficiency of CYP90, a cytochrome P450, controlling cell elongation and de-etiolation in Arabidopsis. Cell 85:171-182. doi:10.1016/S0092-8674(00)81094-6

Wang L, Waltenberger B, Pferschy-Wenzig EM, Blunder M, Liu X, Malainer C, Blazevic T, Schwaiger S, Rollinger JM Heiss EH Schuster D, Kopp B, Bauer R, Stuppner H, Dirsch VM, Atanasov AG (2014) Natural product agonists of peroxisome proliferator-activated receptor gamma (PPARγ): a review. Biochem Pharmacol 92:73-89. doi:10.1016/j.bcp.2014.07.018

Yamamoto R, Fujioka S, Iwamoto K, Demura T, Takatsuto S, Yoshida S, Fukuda H (2007) Co-regulation of brassinosteroid biosynthesisrelated genes during xylem cell differentiation. Plant Cell Physiol 48:74-83. doi:10.1093/pcp/pcl039

Zhu JY, Sae-Seaw J, Wang ZY (2013) Brassinosteroid signaling. Development 140:1615-1620. doi:10.1242/dev.060590

Information

Publisher :KOREAN SOCIETY FOR HORTICULTURAL SCIENCE
Publisher(Ko) :한국원예학회
Journal Title :Horticultural Science and Technology
Journal Title(Ko) :원예과학기술지
Volume : 35
No :5
Pages :628-636
Received Date : 2017-05-18
Revised Date : 2017-07-05
Accepted Date : 2017-07-04
DOI :https://doi.org/10.12972/kjhst.20170067

Horticultural Science and Technology 원예과학기술지 ISSN:1226-8763(Print) 2465-8588(Online)

All Issue