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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front Agric Chin    2011, Vol. 5 Issue (3) : 322-327    https://doi.org/10.1007/s11703-011-1084-4
RESEARCH ARTICLE
Identification of a gene responsible for the 60-day delay in flowering time of Arabidopsis
Jihong XING1, Ye ZHANG1, Jing ZHANG1, Qiaoyun WENG2, Jiao JIA1, Jingao DONG1()
1. Molecular Plant Pathology Lab, Agricultural University of Hebei, Baoding 071001, China; 2. Department of Agricultural and Forest Technology, Hebei North University, Zhangjiakou 075131, China
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Abstract

Identification of genes related to flowering-time in Arabidopsis is very important and meaningful contribution to the flowering process control. One late flowering mutant plant, which exhibits 60-day delay in flowering, was screened from Arabidopsis library of T-DNA insertion. Southern blotting was used to confirm the single copy of exogenetic T-DNA in the genome of the mutant. The flanking sequence of T-DNA insert was obtained by TAIL-PCR and then analyzed by BLAST to confirm that the insertion site locates at the sixth exon of AT2G19520.1 (FVE gene). FVE is considered as a classical flowering time gene in Arabidopsis. It is a component of the autonomous pathway that encodes AtMSI4, which is a putative retinoblastoma-associated protein. The late-flowering mutant is named as fve-4, which is similar to fve-3 of Columbia and allelic with fve-1 and fve-2 of Landsberg erecta. The fve-4 mutant’s delay of flowering was longer than that of fve-3 mutant, whose T-DNA insertion is located at the first exon of FVE gene, suggesting that the sixth exon of FVE gene may play a more important role in the control of floral transition.

Keywords delay of flowering      FVE gene      Arabidopsis      gene identification     
Corresponding Author(s): DONG Jingao,Email:shmdjg@hebau.edu.cn   
Issue Date: 05 September 2011
 Cite this article:   
Jihong XING,Ye ZHANG,Jing ZHANG, et al. Identification of a gene responsible for the 60-day delay in flowering time of Arabidopsis[J]. Front Agric Chin, 2011, 5(3): 322-327.
 URL:  
https://academic.hep.com.cn/fag/EN/10.1007/s11703-011-1084-4
https://academic.hep.com.cn/fag/EN/Y2011/V5/I3/322
PurposePrimersPrimer sequence
Probe preparationN15?-TCGGCTATGACTGGGCACAACAGA-3?
N25?-AAGAAGGCGATAGAAGGCGATGCG-3?
TAIL-PCRLexA25?-CTAATCGCATTATCATCCCCTCG-3?
LexA45?-CTGGTTTTATATACAGCAGTCGACG-3?
LexA55?-AGTCGAGGTAAGATTAGATATGG-3?
AD15?-(AGCT)TCGA(G/C)T(A/T)T(G/C)G(A/T)GTT-3?
PCR identificationM13F5?-CGCCAGGGTTTTCCCAGTCACGAC-3?
M13R5?-AGCGGATAACAATTTCACACAGGA-3?
Cloning of FVEFVEL5?-GAAGAGAGAGAGATATAG-3?
FVER5?-GCACAGAGAAGGAATCATTAGG-3?
RT-PCRFVE-15?-GCTCGGTTTGGTAACAAG-3?
FVE-25?-GTTTCCTCGCTGTAAATC-3?
18S-15?-GTTGCAGTTAAAAAGCTCGT-3?
18S-25?-TTGATTTCTCATAAGGTGCC-3?
Tab.1  Primers used for probe of Southern blotting, TAIL-PCR, PCR, and RT-PCR
PlantNo. of rosette leafDays of flowering
Mutant 221-150±0.3 A86±0.3 A
Col-0 (wild type)12±0.2 B27±0.2 B
Tab.2  The numbers of rosette leaf and flowering days under long-day condition
Fig.1  Phenotype of the wild type (Col-0) and the mutant. A is the mutants screened flower later than Col-0 plants in long days; B is the plant of Col-0 (30 days after sowing); C is the plant of the mutant (30 days after sowing); D is the plant of the mutant (90 days after sowing).
Fig.2  Clone of late flowering-related gene. M: DL2000; 1: the second gene amplification; 2, 3: the third gene amplifications.
Fig.3  Structure of mutant and mutations of the alleles. Boxes denote exons, and lines denote introns.
Fig.4  Conserved domain analysis of FVE.
Fig.5  DNA blot hybridization analysis of the mutant.
Fig.6  The result of PCR of gene. 1: the mutant; 2: wide type; M: Wide Range DNA Marker (100-6000 bp).
Fig.7  The expression of was examined by RT-PCR analysis.
Fig.8  The genetic pathways of flowering in . Positive (arrows) and negative (T-lines) interactions are described. Dotted lines show undescribed interaction (Komeda, 2004).
1 Ausin I, Alonso-Blanco C, Jarillo J A, Ruiz-García L, Martínez-Zapater J M (2004). Regulation of flowering time by FVE, a retinoblastoma-associated protein. Nat Genet , 36(2): 162–166
doi: 10.1038/ng1295 pmid:14745447
2 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(Supp1 l): S18–S31
doi: 10.1105/tpc.015958 pmid:15037730
3 He Y, Michaels S D, Amasino R M (2003). Regulation of flowering time by histone acetylation in Arabidopsis. Science , 302(5651): 1751–1754
doi: 10.1126/science.1091109 pmid:14593187
4 Jang S, Marchal V, Panigrahi K C S, Wenkel S, Soppe W, Deng X W, Valverde F, Coupland G (2008). Arabidopsis COP1 shapes the temporal pattern of CO accumulation conferring a photoperiodic flowering response. EMBO J , 27(8): 1277–1288
doi: 10.1038/emboj.2008.68 pmid:18388858
5 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
doi: 10.1126/science.286.5446.1962 pmid:10583961
6 Kim H J, Hyun Y, Park J Y, Park M J, Park M K, Kim M D, Kim H J, Lee M H, Moon J, Lee I, Kim J (2004). A genetic link between cold responses and flowering time through FVE in Arabidopsis thaliana. Nat Genet , 36(2): 167–171
doi: 10.1038/ng1298 pmid:14745450
7 Kim S, Choi K, Park C, Hwang H J, Lee I (2006). SUPPRESSOR OF FRIGIDA4, encoding a C2H2-Type zinc finger protein, represses flowering by transcriptional activation of Arabidopsis FLOWERING LOCUS C. Plant Cell , 18(11): 2985–2998
doi: 10.1105/tpc.106.045179 pmid:17138694
8 Kole C, Quijada P, Michaels S D, Amasino R M, Osborn T C (2001). Evidence for homology of flowering-time genes VFR2 from Brassica rapa and FLC from Arabidopsis thaliana. Theor Appl Genet , 102(2–3): 425–430
doi: 10.1007/s001220051663
9 Komeda Y (2004). Genetic regulation of time to flower in Arabidopsis thaliana. Annu Rev Plant Biol , 55(1): 521–535
doi: 10.1146/annurev.arplant.55.031903.141644 pmid:15377230
10 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
doi: 10.1101/gad.813600 pmid:10995392
11 Lee I, Aukerman M J, Gore S L, Lohman K N, Michaels S D, Weaver L M, John M C, Feldmann K A, Amasino R M (1994). Isolation of LUMINIDEPENDENS: a gene involved in the control of flowering time in Arabidopsis. Plant Cell , 6(1): 75–83
pmid:7907507
12 Liu Y G, Mitsukawa N, Oosumi T, Whittier R F (1995). Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR. Plant J , 8(3): 457–463
doi: 10.1046/j.1365-313X.1995.08030457.x pmid:7550382
13 Nyathi Y, De Marcos Lousa C, van Roermund C W, Wanders R J, Johnson B, Baldwin S A, Theodoulou F L, Baker A (2010). The Arabidopsis peroxisomal ABC transporter, comatose, complements the Saccharomyces cerevisiae pxa1 pxa2 Δ mutant for metabolism of long-chain fatty acids and exhibits fatty acyl-CoA-stimulated ATPase activity. J Biol Chem , 285(39): 29892–29902
14 Onouchi H, Ige?o 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
pmid:10852935
15 Quesada V, Macknight R, Dean C, Simpson G G (2003). Autoregulation of FCA pre-mRNA processing controls Arabidopsis flowering time. EMBO J , 22(12): 3142–3152
doi: 10.1093/emboj/cdg305 pmid:12805228
16 Schwartz C, Balasubramanian S, Warthmann N, Michael T P, Lempe J, Sureshkumar S, Kobayashi Y, Maloof J N, Borevitz J O, Chory J, Weigel D (2009). Cis-regulatory changes at FLOWERING LOCUS T mediate natural variation in flowering responses of Arabidopsis thaliana. Genetics , 183(2): 723–732
doi: 10.1534/genetics.109.104984 pmid:19652183
17 Simpson G G (2004). The autonomous pathway: epigenetic and post-transcriptional gene regulation in the control of Arabidopsis flowering time. Curr Opin Plant Biol , 7(5): 570–574
doi: 10.1016/j.pbi.2004.07.002 pmid:15337100
18 Simpson G G, Dean C (2002). Arabidopsis, the Rosetta stone of flowering time? Science , 296(5566): 285–289
doi: 10.1126/science.296.5566.285 pmid:11951029
19 Simpson G G, Dijkwel P P, Quesada V, Henderson I, Dean C (2003). FY is an RNA 3? end-processing factor that interacts with FCA to control the Arabidopsis floral transition. Cell , 113(6): 777–787
doi: 10.1016/S0092-8674(03)00425-2 pmid:12809608
20 Tadege M, Sheldon C C, Helliwell C A, Stoutjesdijk P, Dennis E S, Peacock W J (2001). Control of flowering time by FLC orthologues in Brassica napus. Plant J , 28(5): 545–553
doi: 10.1046/j.1365-313X.2001.01182.x pmid:11849594
21 Waters B M, Chu H-H, DiDonato R J, Roberts L A, Eisley R B, Lahner B, Salt D E, Walker E L (2006). Mutations in Arabidopsis yellow stripe-like1 and yellow stripe-like3 reveal their roles in metal ion homeostasis and loading of metal ions in seeds. Plant Physiol , 141(4): 1446–1458 .
22 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
doi: 10.1016/0092-8674(92)90295-N pmid:1350515
23 Yan Z Q, Liang D W, Liu H, Zheng G C (2010). FLC: A key regulator of flowering time in Arabidopsis. Russ J Plant Physiol , 57(2): 166–174
doi: 10.1134/S1021443710020020
24 Zhang J, Xu J X, Kong Y Z, Ji Z D, Wang X C, An F Y, Li C, Sun J Q, Zhang S Z, Yang X H, Mu J Y, Liu X F, Li J Y, Xue Y B, Zuo J R (2005). Generation of chemical-inducible activation tagging T-DNA insertion lines of Arabidopsis thaliana. Acta Genetica Sinica , 32(10): 1082–1088 (in Chinese)
pmid:16252704
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