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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

Expression analysis of related genes from 35S::CgFT transgenic plants under 12 days LD (16 h L/8 h D) conditions. The amount of TUBULIN transcripts in Arabidopsis thaliana was used as an internal control. 1–4 represent four lines (3-4, 7-8, 30-10, 9-5) of 35S::CgFT transgenic plants. The annealing temperature in PCR was 58 °C for TUBULIN, CgFT and AtAP1. 25 cycles for TUBULIN, 28 cycles for CgFT and AtAP1. The materials were measured using leaves of WT and transgenic Arabidopsis thaliana plants.
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f6-ijms-13-11385: Expression analysis of related genes from 35S::CgFT transgenic plants under 12 days LD (16 h L/8 h D) conditions. The amount of TUBULIN transcripts in Arabidopsis thaliana was used as an internal control. 1–4 represent four lines (3-4, 7-8, 30-10, 9-5) of 35S::CgFT transgenic plants. The annealing temperature in PCR was 58 °C for TUBULIN, CgFT and AtAP1. 25 cycles for TUBULIN, 28 cycles for CgFT and AtAP1. The materials were measured using leaves of WT and transgenic Arabidopsis thaliana plants.

Mentions: To explore whether the early flowering phenotype was correlated with CgFT expression in 35S::CgFT transgenic plants, RT-PCR analysis was performed. As shown in Figure 6, higher CgFT expression was observed in the severe 35S::CgFT transgenic plants more than in the transgenic plants with a less severe or WT phenotype. Further analysis indicated that the promotion of flowering time in severe early flowering 35S::CgFT transgenic plants was also related to significant up-regulation of the flower meristem identity gene AtAP1 in transgenic plants (Figure 6). This result indicates that the function of Cymbidium FT is similar to that of A. thaliana FT in regulating flowering time.


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)

Expression analysis of related genes from 35S::CgFT transgenic plants under 12 days LD (16 h L/8 h D) conditions. The amount of TUBULIN transcripts in Arabidopsis thaliana was used as an internal control. 1–4 represent four lines (3-4, 7-8, 30-10, 9-5) of 35S::CgFT transgenic plants. The annealing temperature in PCR was 58 °C for TUBULIN, CgFT and AtAP1. 25 cycles for TUBULIN, 28 cycles for CgFT and AtAP1. The materials were measured using leaves of WT and transgenic Arabidopsis thaliana plants.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f6-ijms-13-11385: Expression analysis of related genes from 35S::CgFT transgenic plants under 12 days LD (16 h L/8 h D) conditions. The amount of TUBULIN transcripts in Arabidopsis thaliana was used as an internal control. 1–4 represent four lines (3-4, 7-8, 30-10, 9-5) of 35S::CgFT transgenic plants. The annealing temperature in PCR was 58 °C for TUBULIN, CgFT and AtAP1. 25 cycles for TUBULIN, 28 cycles for CgFT and AtAP1. The materials were measured using leaves of WT and transgenic Arabidopsis thaliana plants.
Mentions: To explore whether the early flowering phenotype was correlated with CgFT expression in 35S::CgFT transgenic plants, RT-PCR analysis was performed. As shown in Figure 6, higher CgFT expression was observed in the severe 35S::CgFT transgenic plants more than in the transgenic plants with a less severe or WT phenotype. Further analysis indicated that the promotion of flowering time in severe early flowering 35S::CgFT transgenic plants was also related to significant up-regulation of the flower meristem identity gene AtAP1 in transgenic plants (Figure 6). This result indicates that the function of Cymbidium FT is similar to that of A. thaliana FT in regulating flowering time.

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