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Molecular cloning and functional analysis of Three FLOWERING LOCUS T (FT) homologous genes from Chinese Cymbidium.

Huang W, Fang Z, Zeng S, Zhang J, Wu K, Chen Z, Teixeira da Silva JA, Duan J - Int J Mol Sci (2012)

Bottom Line: Alignment of the AA sequences revealed that CsFT, CgFT and CeFT contain a conserved domain, which is characteristic of the PEBP-RKIP superfamily, and which share high identity with FT of other plants in GenBank: 94% with OnFT from Oncidium Gower Ramsey, 79% with Hd3a from Oryza sativa, and 74% with FT from Arabidopsis thaliana. qRT-PCR analysis showed a diurnal expression pattern of CsFT, CgFT and CeFT following both long day (LD, 16-h light/8-h dark) and short day (SD, 8-h light/16-h dark) treatment.While the transcripts of both CsFT and CeFT under LD were significantly higher than under SD, those of CgFT were higher under SD.Our data indicates that CgFT is a putative phosphatidylethanolamine-binding protein gene in Cymbidium that may regulate the vegetative to reproductive transition in flowers, similar to its Arabidopsis ortholog.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of South China Agricultural Plant Genetics and Breeding, South China Botanical Garden, The Chinese Academy of Sciences, Guangzhou 510650, China; E-Mails: weitingpink@hotmail.com (W.H.); zmfang88@163.com (Z.F.); cymbidium1979@yahoo.com.cn (J.Z.); wu_kunlin@163.com (K.W.); duanj@scib.ac.cn (J.D.) ; Graduate University of the Chinese Academy of Sciences, Beijing 100049, China.

ABSTRACT
The FLOWERING LOCUS T (FT) gene plays crucial roles in regulating the transition from the vegetative to reproductive phase. To understand the molecular mechanism of reproduction, three homologous FT genes were isolated and characterized from Cymbidium sinense "Qi Jian Bai Mo", Cymbidium goeringii and Cymbidium ensifolium "Jin Si Ma Wei". The three genes contained 618-bp nucleotides with a 531-bp open reading frame (ORF) of encoding 176 amino acids (AAs). Alignment of the AA sequences revealed that CsFT, CgFT and CeFT contain a conserved domain, which is characteristic of the PEBP-RKIP superfamily, and which share high identity with FT of other plants in GenBank: 94% with OnFT from Oncidium Gower Ramsey, 79% with Hd3a from Oryza sativa, and 74% with FT from Arabidopsis thaliana. qRT-PCR analysis showed a diurnal expression pattern of CsFT, CgFT and CeFT following both long day (LD, 16-h light/8-h dark) and short day (SD, 8-h light/16-h dark) treatment. While the transcripts of both CsFT and CeFT under LD were significantly higher than under SD, those of CgFT were higher under SD. Ectopic expression of CgFT in transgenic Arabidopsis plants resulted in early flowering compared to wild-type plants and significant up-regulation of APETALA1 (AP1) expression. Our data indicates that CgFT is a putative phosphatidylethanolamine-binding protein gene in Cymbidium that may regulate the vegetative to reproductive transition in flowers, similar to its Arabidopsis ortholog.

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Related in: MedlinePlus

Diurnal expression pattern of FT in long days and short days. Expression pattern of (A) CsFT (Cymbidium sinense); (B) CgFT (Cymbidium goeringii); (C) CeFT (Cymbidium ensifolium). LD is marked by open circles. SD is marked by closed circles. Error bars depict SD (n = 4). The materials were measured using non-annual leaves of good condition.
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f4-ijms-13-11385: Diurnal expression pattern of FT in long days and short days. Expression pattern of (A) CsFT (Cymbidium sinense); (B) CgFT (Cymbidium goeringii); (C) CeFT (Cymbidium ensifolium). LD is marked by open circles. SD is marked by closed circles. Error bars depict SD (n = 4). The materials were measured using non-annual leaves of good condition.

Mentions: To explore whether the expression of CsFT, CgFT, and CeFT was influenced by daily oscillations in photoperiod, the expression of CsFT, CgFT, and CeFT mRNA was analyzed every 4 h over a 24-h period in long day (LD) and short day (SD) conditions using quantitative real time PCR (Figure 4). CsFT expression (Figure 4A) was different under SD and LD: it was higher under LD than under SD. Expression of CsFT showed an increasing trend under LD, indicating that expression of CsFT was very sensitive to changes in light under LD. CgFT (Figure 4B) was also regulated by light but its expression was significantly higher under SD than under LD, but there was a similar change in pattern under both conditions, with the level highest at the 4th hour of the light period and lowest at the 16th hour. C. ensifolium “Jin Si Ma Wei” CeFT (Figure 4C) showed higher expression under LD than SD with a rhythmic cycle, but peaked at different times (12 h and 36 h of LD, 20 h and 36 h of SD). CeFT mRNA showed highest levels of expression 12 h into the light period and lowest levels at dawn. CgFT expression was thus different from CsFT and CeFT expression and all were regulated by light.


Molecular cloning and functional analysis of Three FLOWERING LOCUS T (FT) homologous genes from Chinese Cymbidium.

Huang W, Fang Z, Zeng S, Zhang J, Wu K, Chen Z, Teixeira da Silva JA, Duan J - Int J Mol Sci (2012)

Diurnal expression pattern of FT in long days and short days. Expression pattern of (A) CsFT (Cymbidium sinense); (B) CgFT (Cymbidium goeringii); (C) CeFT (Cymbidium ensifolium). LD is marked by open circles. SD is marked by closed circles. Error bars depict SD (n = 4). The materials were measured using non-annual leaves of good condition.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3472752&req=5

f4-ijms-13-11385: Diurnal expression pattern of FT in long days and short days. Expression pattern of (A) CsFT (Cymbidium sinense); (B) CgFT (Cymbidium goeringii); (C) CeFT (Cymbidium ensifolium). LD is marked by open circles. SD is marked by closed circles. Error bars depict SD (n = 4). The materials were measured using non-annual leaves of good condition.
Mentions: To explore whether the expression of CsFT, CgFT, and CeFT was influenced by daily oscillations in photoperiod, the expression of CsFT, CgFT, and CeFT mRNA was analyzed every 4 h over a 24-h period in long day (LD) and short day (SD) conditions using quantitative real time PCR (Figure 4). CsFT expression (Figure 4A) was different under SD and LD: it was higher under LD than under SD. Expression of CsFT showed an increasing trend under LD, indicating that expression of CsFT was very sensitive to changes in light under LD. CgFT (Figure 4B) was also regulated by light but its expression was significantly higher under SD than under LD, but there was a similar change in pattern under both conditions, with the level highest at the 4th hour of the light period and lowest at the 16th hour. C. ensifolium “Jin Si Ma Wei” CeFT (Figure 4C) showed higher expression under LD than SD with a rhythmic cycle, but peaked at different times (12 h and 36 h of LD, 20 h and 36 h of SD). CeFT mRNA showed highest levels of expression 12 h into the light period and lowest levels at dawn. CgFT expression was thus different from CsFT and CeFT expression and all were regulated by light.

Bottom Line: Alignment of the AA sequences revealed that CsFT, CgFT and CeFT contain a conserved domain, which is characteristic of the PEBP-RKIP superfamily, and which share high identity with FT of other plants in GenBank: 94% with OnFT from Oncidium Gower Ramsey, 79% with Hd3a from Oryza sativa, and 74% with FT from Arabidopsis thaliana. qRT-PCR analysis showed a diurnal expression pattern of CsFT, CgFT and CeFT following both long day (LD, 16-h light/8-h dark) and short day (SD, 8-h light/16-h dark) treatment.While the transcripts of both CsFT and CeFT under LD were significantly higher than under SD, those of CgFT were higher under SD.Our data indicates that CgFT is a putative phosphatidylethanolamine-binding protein gene in Cymbidium that may regulate the vegetative to reproductive transition in flowers, similar to its Arabidopsis ortholog.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of South China Agricultural Plant Genetics and Breeding, South China Botanical Garden, The Chinese Academy of Sciences, Guangzhou 510650, China; E-Mails: weitingpink@hotmail.com (W.H.); zmfang88@163.com (Z.F.); cymbidium1979@yahoo.com.cn (J.Z.); wu_kunlin@163.com (K.W.); duanj@scib.ac.cn (J.D.) ; Graduate University of the Chinese Academy of Sciences, Beijing 100049, China.

ABSTRACT
The FLOWERING LOCUS T (FT) gene plays crucial roles in regulating the transition from the vegetative to reproductive phase. To understand the molecular mechanism of reproduction, three homologous FT genes were isolated and characterized from Cymbidium sinense "Qi Jian Bai Mo", Cymbidium goeringii and Cymbidium ensifolium "Jin Si Ma Wei". The three genes contained 618-bp nucleotides with a 531-bp open reading frame (ORF) of encoding 176 amino acids (AAs). Alignment of the AA sequences revealed that CsFT, CgFT and CeFT contain a conserved domain, which is characteristic of the PEBP-RKIP superfamily, and which share high identity with FT of other plants in GenBank: 94% with OnFT from Oncidium Gower Ramsey, 79% with Hd3a from Oryza sativa, and 74% with FT from Arabidopsis thaliana. qRT-PCR analysis showed a diurnal expression pattern of CsFT, CgFT and CeFT following both long day (LD, 16-h light/8-h dark) and short day (SD, 8-h light/16-h dark) treatment. While the transcripts of both CsFT and CeFT under LD were significantly higher than under SD, those of CgFT were higher under SD. Ectopic expression of CgFT in transgenic Arabidopsis plants resulted in early flowering compared to wild-type plants and significant up-regulation of APETALA1 (AP1) expression. Our data indicates that CgFT is a putative phosphatidylethanolamine-binding protein gene in Cymbidium that may regulate the vegetative to reproductive transition in flowers, similar to its Arabidopsis ortholog.

Show MeSH
Related in: MedlinePlus