Frontiers of Agriculture in China >
Construction of RNAi vector for flower-related gene and verifications of the mutant in Arabidopsis thaliana
Received date: 10 Jan 2011
Accepted date: 09 Mar 2011
Published date: 05 Sep 2011
Copyright
The timing of floral transition is tightly controlled by a combination of endogenous and environmental signals. One early flowering mutant plant was screened from Arabidopsis library of T-DNA insertion to accelerate flowering under short-day condition, and a related-gene EFS1 (AT4G36680.1) was isolated and identified as a novel flowering-time gene of Arabidopsis in our preliminary studies. To investigate the function and the specific mechanism of EFS1 in the flower process control, the RNAi expression vector containing EFS1 gene-specific sequences in the sense and antisense orientations was constructed and transferred into Arabidopsis by using the floral-dip method, with 11 transgenic plants obtained through hygromycin B screening and PCR assays. The results showed that the expression level of EFS1 in transgenic lines was significantly lower than that in wild type and efs1 mutant. The flowering time of the efs1 mutant and RNAi transgenic plants was much earlier than that of wild-type plants. This result further verified that the EFS1 gene played an important role in flowering, and its specific mechanisms need further study. These work provided a foundation to further regulatory mechanisms of EFS1 in the control of floral transition.
Key words： Arabidopsis thaliana; RNAi; EFS1 gene; early flowering
Jing ZHANG , Jihong XING , Jingao DONG . Construction of RNAi vector for flower-related gene and verifications of the mutant in Arabidopsis thaliana[J]. Frontiers of Agriculture in China, 2011 , 5(3) : 333 -337 . DOI: 10.1007/s11703-011-1094-2
| 1 |
Boss P K, Bastow R M, Mylne J S, Dean C (2004). Multiple pathways in the decision to flower: enabling, promoting, and resetting. Plant Cell, 16(suppl1): S18–S31
|
| 2 |
Boutros M, Kiger A A, Armknecht S, Kerr K, Hild M, Koch B, Haas S A, Paro R, Perrimon N, Perrimon N (2004). Genome-wide RNAi analysis of growth and viability in Drosophila cells. Science, 303(5659): 832–835
|
| 3 |
Fire A, Xu S, Montgomery M K, Kostas S A, Driver S E, Mello C C (1998). Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 391(6669): 806–811
|
| 4 |
Frantz S (2003). Studies reveal potential pitfalls of RNAi. Nature Reviews Drug Discovery, 2(10): 763–764
|
| 5 |
Hannon G J (2002). RNA interference. Nature, 418(6894): 244–251
|
| 6 |
Hunter C, Poethig R S (2003). miSSING LINKS: miRNAs and plant development. Curr Opin Genet Dev, 13(4): 372–378
|
| 7 |
Kamath R S, Fraser A G, Dong Y, Poulin G, Durbin R, Gotta M, Kanapin A, Le Bot N, Moreno S, Sohrmann M, Welchman D P, Zipperlen P, Ahringer J (2003). Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature, 421(6920): 231–237
|
| 8 |
Kardailsky I, Shukla V K, Ahn J H, Dagenais N, Christensen S K, Nguyen J T, Chory J, Harrison M J, Weigel D (1999). Activation tagging of the floral inducer FT. Science, 286(5446): 1962–1965
|
| 9 |
Kidner C A, Martienssen R A (2003). Macro effects of microRNAs in plants. Trends Genet, 19(1): 13–16
|
| 10 |
Komeda Y (2004). Genetic regulation of time to flower in Arabidopsis thaliana. Annu Rev Plant Biol, 55(1): 521–535
|
| 11 |
Lee H, Suh S S, Park E, Cho E, Ahn J H, Kim S G, Lee J S, Kwon Y M, Lee I (2000). The AGAMOUS-LIKE 20 MADS domain protein integrates floral inductive pathways in Arabidopsis. Genes Dev, 14(18): 2366–2376
|
| 12 |
Onouchi H, Igeño M I, Périlleux C, Graves K, Coupland G (2000). Mutagenesis of plants overexpressing CONSTANS demonstrates novel interactions among Arabidopsis flowering-time genes. Plant Cell, 12(6): 885–900
|
| 13 |
Pickford A S, Cogoni C (2003). RNA-mediated gene silencing. Cellular and Molecular Life Sciences, 60(5): 871–882
|
| 14 |
Putterill J, Laurie R, Macknight R (2004). It’s time to flower: the genetic control of flowering time. Bioessays, 26(4): 363–373
|
| 15 |
Simpson G G, Dean C (2002). Arabidopsis, the Rosetta stone of flowering time? Science, 296(5566): 285–289
|
| 16 |
Tijsterman M, Ketting R F, Plasterk R H (2002). The genetics of RNA silencing. Annu Rev Genet, 36(1): 489–519
|
| 17 |
Waterhouse P, Graham M, Wang M B (1998). Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. PNAS, 95: 13959–13964
|
| 18 |
Weigel D, Alvarez J, Smyth D R, Yanofsky M F, Meyerowitz E M (1992). LEAFY controls floral meristem identity in Arabidopsis. Cell, 69(5): 843–859
|
/
| 〈 |
|
〉 |