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Overexpression of two PsnAP1 genes from Populus simonii × P. nigra causes early flowering in transgenic tobacco and Arabidopsis.

Zheng T, Li S, Zang L, Dai L, Yang C, Qu GZ - PLoS ONE (2014)

Bottom Line: Overexpression of PsnAP1-1 and PsnAP1-2 in tobacco under the control of a CaMV 35S promoter significantly enhanced early flowering.These transgenic plants also showed much earlier stem initiation and higher rates of photosynthesis than did wild-type tobacco. qRT-PCR analysis further indicated that overexpression of PsnAP1-1 and PsnAP1-2 resulted in up-regulation of genes related to flowering, such as NtMADS4, NtMADS5 and NtMADS11.Overexpression of PsnAP1-1 and PsnAP1-2 in Arabidopsis also induced early flowering, but did not complement the ap1-10 floral morphology to any noticeable extent.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Tree Genetics and Breeding (Northeast Forestry University), Harbin, China.

ABSTRACT
In Arabidopsis, AP1 is a floral meristem identity gene and plays an important role in floral organ development. In this study, PsnAP1-1 and PsnAP1-2 were isolated from the male reproductive buds of poplar (Populus simonii × P. nigra), which are the orthologs of AP1 in Arabidopsis, by sequence analysis. Northern blot and qRT-PCR analysis showed that PsnAP1-1 and PsnAP1-2 exhibited high expression level in early inflorescence development of poplar. Subcellular localization showed the PsnAP1-1 and PsnAP1-2 proteins are localized in the nucleus. Overexpression of PsnAP1-1 and PsnAP1-2 in tobacco under the control of a CaMV 35S promoter significantly enhanced early flowering. These transgenic plants also showed much earlier stem initiation and higher rates of photosynthesis than did wild-type tobacco. qRT-PCR analysis further indicated that overexpression of PsnAP1-1 and PsnAP1-2 resulted in up-regulation of genes related to flowering, such as NtMADS4, NtMADS5 and NtMADS11. Overexpression of PsnAP1-1 and PsnAP1-2 in Arabidopsis also induced early flowering, but did not complement the ap1-10 floral morphology to any noticeable extent. This study indicates that PsnAP1-1 and PsnAP1-2 play a role in floral transition of poplar.

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Overexpression of PsnAP1-1 and PsnAP1-2 in Arabidopsis transgenic lines.(A) Comparison of a wild-type Columbia, 35S::PsnAP1-1, and 35S::PsnAP1-2 20 d after planting under short-day condition. (B) Comparison of a wild-type Columbia and 35S::PsnAP1-1 at about 20 d after planting under long-day conditions, left wild-type, right 35S::PsnAP1-1 transgenic plants, the arrows represent the flowering seedlings. (C) Close-up of wild-type 20 d after planting under short-day conditions. (D) One 35S::PsnAP1-1 T1 transgenic seedling flowered too early for the seeds to be harvested. (E) One 35S::PsnAP1-2 T1 transgenic seedling flowered too early for the seeds to be harvested. (F) Wild-type 45 d after planting under short-day conditions. (G) A 35S::PsnAP1-2 line with the formation of terminal flowers. (H) A 35S::PsnAP1-2 line with deformed flowers. (I) A 35S::PsnAP1-1 line with curling leaves and inflorescence branches converted to solitary flowers. (J, K, L) Close-up of terminal flowers (J), deformed flowers (K), and apical and lateral shoots developing as flowers (L) in 35S::PsnAP1 lines. WT wild-type, PsnAP1-1 35S::PsnAP1-1 line in wild-type, PsnAP1-2 35S::PsnAP1-2 line in wild-type. The arrows represent the locations of observation.
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pone-0111725-g010: Overexpression of PsnAP1-1 and PsnAP1-2 in Arabidopsis transgenic lines.(A) Comparison of a wild-type Columbia, 35S::PsnAP1-1, and 35S::PsnAP1-2 20 d after planting under short-day condition. (B) Comparison of a wild-type Columbia and 35S::PsnAP1-1 at about 20 d after planting under long-day conditions, left wild-type, right 35S::PsnAP1-1 transgenic plants, the arrows represent the flowering seedlings. (C) Close-up of wild-type 20 d after planting under short-day conditions. (D) One 35S::PsnAP1-1 T1 transgenic seedling flowered too early for the seeds to be harvested. (E) One 35S::PsnAP1-2 T1 transgenic seedling flowered too early for the seeds to be harvested. (F) Wild-type 45 d after planting under short-day conditions. (G) A 35S::PsnAP1-2 line with the formation of terminal flowers. (H) A 35S::PsnAP1-2 line with deformed flowers. (I) A 35S::PsnAP1-1 line with curling leaves and inflorescence branches converted to solitary flowers. (J, K, L) Close-up of terminal flowers (J), deformed flowers (K), and apical and lateral shoots developing as flowers (L) in 35S::PsnAP1 lines. WT wild-type, PsnAP1-1 35S::PsnAP1-1 line in wild-type, PsnAP1-2 35S::PsnAP1-2 line in wild-type. The arrows represent the locations of observation.

Mentions: We next overexpressed the PsnAP1-1 and PsnAP1-2 in Arabidopsis as well as in ap1 mutant. The constitutive expression of PsnAP1-1 and PsnAP1-2 in the wild-type and ap1-10 mutant plants was verified using Northern blot analysis (Fig. 4E, F). The results showed that overexpression of PsnAP1-1 and PsnAP1-2 reduced the vegetative phase of the life cycle of the wild type. Under long-day growth conditions, flowers were first visible on most transgenic lines after approximately 10 d, however, the wild-type plants flowered after an average of approximately 25 d (Table 1, Fig. 10A, B, C). In addition, few T1 transgenic seedlings flowered very early so that they were too fragile to harvest seeds (Fig. 10D, E). However, these T1 transformants were dwarfed, with final aerial heights less than 5 cm, and wild-type showed final aerial height more than 30 cm after more than 60 d of growth (Fig. 10F, G, H, I). A few transgenic lines showed fused terminal flowers, curled leaves, deformed flowers, and lateral shoots that developed into solitary flowers (Fig. 10G–L).


Overexpression of two PsnAP1 genes from Populus simonii × P. nigra causes early flowering in transgenic tobacco and Arabidopsis.

Zheng T, Li S, Zang L, Dai L, Yang C, Qu GZ - PLoS ONE (2014)

Overexpression of PsnAP1-1 and PsnAP1-2 in Arabidopsis transgenic lines.(A) Comparison of a wild-type Columbia, 35S::PsnAP1-1, and 35S::PsnAP1-2 20 d after planting under short-day condition. (B) Comparison of a wild-type Columbia and 35S::PsnAP1-1 at about 20 d after planting under long-day conditions, left wild-type, right 35S::PsnAP1-1 transgenic plants, the arrows represent the flowering seedlings. (C) Close-up of wild-type 20 d after planting under short-day conditions. (D) One 35S::PsnAP1-1 T1 transgenic seedling flowered too early for the seeds to be harvested. (E) One 35S::PsnAP1-2 T1 transgenic seedling flowered too early for the seeds to be harvested. (F) Wild-type 45 d after planting under short-day conditions. (G) A 35S::PsnAP1-2 line with the formation of terminal flowers. (H) A 35S::PsnAP1-2 line with deformed flowers. (I) A 35S::PsnAP1-1 line with curling leaves and inflorescence branches converted to solitary flowers. (J, K, L) Close-up of terminal flowers (J), deformed flowers (K), and apical and lateral shoots developing as flowers (L) in 35S::PsnAP1 lines. WT wild-type, PsnAP1-1 35S::PsnAP1-1 line in wild-type, PsnAP1-2 35S::PsnAP1-2 line in wild-type. The arrows represent the locations of observation.
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Related In: Results  -  Collection

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pone-0111725-g010: Overexpression of PsnAP1-1 and PsnAP1-2 in Arabidopsis transgenic lines.(A) Comparison of a wild-type Columbia, 35S::PsnAP1-1, and 35S::PsnAP1-2 20 d after planting under short-day condition. (B) Comparison of a wild-type Columbia and 35S::PsnAP1-1 at about 20 d after planting under long-day conditions, left wild-type, right 35S::PsnAP1-1 transgenic plants, the arrows represent the flowering seedlings. (C) Close-up of wild-type 20 d after planting under short-day conditions. (D) One 35S::PsnAP1-1 T1 transgenic seedling flowered too early for the seeds to be harvested. (E) One 35S::PsnAP1-2 T1 transgenic seedling flowered too early for the seeds to be harvested. (F) Wild-type 45 d after planting under short-day conditions. (G) A 35S::PsnAP1-2 line with the formation of terminal flowers. (H) A 35S::PsnAP1-2 line with deformed flowers. (I) A 35S::PsnAP1-1 line with curling leaves and inflorescence branches converted to solitary flowers. (J, K, L) Close-up of terminal flowers (J), deformed flowers (K), and apical and lateral shoots developing as flowers (L) in 35S::PsnAP1 lines. WT wild-type, PsnAP1-1 35S::PsnAP1-1 line in wild-type, PsnAP1-2 35S::PsnAP1-2 line in wild-type. The arrows represent the locations of observation.
Mentions: We next overexpressed the PsnAP1-1 and PsnAP1-2 in Arabidopsis as well as in ap1 mutant. The constitutive expression of PsnAP1-1 and PsnAP1-2 in the wild-type and ap1-10 mutant plants was verified using Northern blot analysis (Fig. 4E, F). The results showed that overexpression of PsnAP1-1 and PsnAP1-2 reduced the vegetative phase of the life cycle of the wild type. Under long-day growth conditions, flowers were first visible on most transgenic lines after approximately 10 d, however, the wild-type plants flowered after an average of approximately 25 d (Table 1, Fig. 10A, B, C). In addition, few T1 transgenic seedlings flowered very early so that they were too fragile to harvest seeds (Fig. 10D, E). However, these T1 transformants were dwarfed, with final aerial heights less than 5 cm, and wild-type showed final aerial height more than 30 cm after more than 60 d of growth (Fig. 10F, G, H, I). A few transgenic lines showed fused terminal flowers, curled leaves, deformed flowers, and lateral shoots that developed into solitary flowers (Fig. 10G–L).

Bottom Line: Overexpression of PsnAP1-1 and PsnAP1-2 in tobacco under the control of a CaMV 35S promoter significantly enhanced early flowering.These transgenic plants also showed much earlier stem initiation and higher rates of photosynthesis than did wild-type tobacco. qRT-PCR analysis further indicated that overexpression of PsnAP1-1 and PsnAP1-2 resulted in up-regulation of genes related to flowering, such as NtMADS4, NtMADS5 and NtMADS11.Overexpression of PsnAP1-1 and PsnAP1-2 in Arabidopsis also induced early flowering, but did not complement the ap1-10 floral morphology to any noticeable extent.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Tree Genetics and Breeding (Northeast Forestry University), Harbin, China.

ABSTRACT
In Arabidopsis, AP1 is a floral meristem identity gene and plays an important role in floral organ development. In this study, PsnAP1-1 and PsnAP1-2 were isolated from the male reproductive buds of poplar (Populus simonii × P. nigra), which are the orthologs of AP1 in Arabidopsis, by sequence analysis. Northern blot and qRT-PCR analysis showed that PsnAP1-1 and PsnAP1-2 exhibited high expression level in early inflorescence development of poplar. Subcellular localization showed the PsnAP1-1 and PsnAP1-2 proteins are localized in the nucleus. Overexpression of PsnAP1-1 and PsnAP1-2 in tobacco under the control of a CaMV 35S promoter significantly enhanced early flowering. These transgenic plants also showed much earlier stem initiation and higher rates of photosynthesis than did wild-type tobacco. qRT-PCR analysis further indicated that overexpression of PsnAP1-1 and PsnAP1-2 resulted in up-regulation of genes related to flowering, such as NtMADS4, NtMADS5 and NtMADS11. Overexpression of PsnAP1-1 and PsnAP1-2 in Arabidopsis also induced early flowering, but did not complement the ap1-10 floral morphology to any noticeable extent. This study indicates that PsnAP1-1 and PsnAP1-2 play a role in floral transition of poplar.

Show MeSH