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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 ap1-10 mutant Arabidopsis caused early flowering.(A) The flowers of wild-type Columbia. (B) Seedlings of ap1-10 mutant 20 d after planting under short-day conditions. (C) The flowers of ap1-10 mutant. (D) Flowers of representative 35S::PsnAP1-1 in ap1-10 plants. (E) The flowers of representative 35S::PsnAP1-2 in ap1-10 plant. (F) Control ap1-10 at 45 d after planting under short-day conditions. (G) A 35S::PsnAP1-1 transgenic line in ap1-10 flowered under short-day growth conditions. (H) A 35S::PsnAP1-2 transgenic line in ap1-10 flowered under short-day growth conditions. (I, J) A 35S::PsnAP1-2 line in ap1-10 showing formation of terminal flowers (I) and curled leaves (J). (K, L) Two 35S::PsnAP1-1 lines in ap1-10 showed curled bracts subtending the solitary flowers. (M) A 35S::PsnAP1-2 line with conversion of secondary inflorescence branches to solitary flowers. WT wild-type, PsnAP1-1 35S::PsnAP1-1 line in ap1-10 mutant, PsnAP1-2 35S::PsnAP1-2 line in ap1-10 mutant. The arrows represent the locations of observation.
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pone-0111725-g011: Overexpression of PsnAP1-1 and PsnAP1-2 in ap1-10 mutant Arabidopsis caused early flowering.(A) The flowers of wild-type Columbia. (B) Seedlings of ap1-10 mutant 20 d after planting under short-day conditions. (C) The flowers of ap1-10 mutant. (D) Flowers of representative 35S::PsnAP1-1 in ap1-10 plants. (E) The flowers of representative 35S::PsnAP1-2 in ap1-10 plant. (F) Control ap1-10 at 45 d after planting under short-day conditions. (G) A 35S::PsnAP1-1 transgenic line in ap1-10 flowered under short-day growth conditions. (H) A 35S::PsnAP1-2 transgenic line in ap1-10 flowered under short-day growth conditions. (I, J) A 35S::PsnAP1-2 line in ap1-10 showing formation of terminal flowers (I) and curled leaves (J). (K, L) Two 35S::PsnAP1-1 lines in ap1-10 showed curled bracts subtending the solitary flowers. (M) A 35S::PsnAP1-2 line with conversion of secondary inflorescence branches to solitary flowers. WT wild-type, PsnAP1-1 35S::PsnAP1-1 line in ap1-10 mutant, PsnAP1-2 35S::PsnAP1-2 line in ap1-10 mutant. The arrows represent the locations of observation.

Mentions: To further test the function of PsnAP1-1 and PsnAP1-2, both ORF sequences, which were under a control of a 35S promoter, were introduced into Arabidopsis ap1-10 mutant plants. In the Columbia background, ap1-10 is a strong allele that causes sepals to convert to bract-like structures. Analysis of T3 lines showed that the ap1-10 plants overexpressed with the PsnAP1-1 or PsnAP1-2 were morphologically distinguishable from control plants. However, overexpression of PsnAP1 didn’t complement the ap1-10 phenotype (Fig. 11C–E). The 35S::PsnAP1 ap1-10 plants had reduced height, exhibited less branching, and terminated growth prematurely, producing one ap1-10 like flower each (Fig. 11F, K–M). Flowering time and the number of rosette leaves number were significantly less in the transgenic ap1-10 plants than those in non-transgenic ap1-10 plants (Table 1). Under short-day growth conditions, ap1-10 plants overexpressed with the PsnAP1-1 or PsnAP1-2 also flowered after an average of 10 d, but no visible inflorescence shoots were detected on the original ap1-10 mutants (Fig. 11B, G, H). A few transgenic lines also showed fused terminal flower, curled leaves, and solitary flowers (Fig. 11I, G, K–M).


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 ap1-10 mutant Arabidopsis caused early flowering.(A) The flowers of wild-type Columbia. (B) Seedlings of ap1-10 mutant 20 d after planting under short-day conditions. (C) The flowers of ap1-10 mutant. (D) Flowers of representative 35S::PsnAP1-1 in ap1-10 plants. (E) The flowers of representative 35S::PsnAP1-2 in ap1-10 plant. (F) Control ap1-10 at 45 d after planting under short-day conditions. (G) A 35S::PsnAP1-1 transgenic line in ap1-10 flowered under short-day growth conditions. (H) A 35S::PsnAP1-2 transgenic line in ap1-10 flowered under short-day growth conditions. (I, J) A 35S::PsnAP1-2 line in ap1-10 showing formation of terminal flowers (I) and curled leaves (J). (K, L) Two 35S::PsnAP1-1 lines in ap1-10 showed curled bracts subtending the solitary flowers. (M) A 35S::PsnAP1-2 line with conversion of secondary inflorescence branches to solitary flowers. WT wild-type, PsnAP1-1 35S::PsnAP1-1 line in ap1-10 mutant, PsnAP1-2 35S::PsnAP1-2 line in ap1-10 mutant. The arrows represent the locations of observation.
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Related In: Results  -  Collection

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pone-0111725-g011: Overexpression of PsnAP1-1 and PsnAP1-2 in ap1-10 mutant Arabidopsis caused early flowering.(A) The flowers of wild-type Columbia. (B) Seedlings of ap1-10 mutant 20 d after planting under short-day conditions. (C) The flowers of ap1-10 mutant. (D) Flowers of representative 35S::PsnAP1-1 in ap1-10 plants. (E) The flowers of representative 35S::PsnAP1-2 in ap1-10 plant. (F) Control ap1-10 at 45 d after planting under short-day conditions. (G) A 35S::PsnAP1-1 transgenic line in ap1-10 flowered under short-day growth conditions. (H) A 35S::PsnAP1-2 transgenic line in ap1-10 flowered under short-day growth conditions. (I, J) A 35S::PsnAP1-2 line in ap1-10 showing formation of terminal flowers (I) and curled leaves (J). (K, L) Two 35S::PsnAP1-1 lines in ap1-10 showed curled bracts subtending the solitary flowers. (M) A 35S::PsnAP1-2 line with conversion of secondary inflorescence branches to solitary flowers. WT wild-type, PsnAP1-1 35S::PsnAP1-1 line in ap1-10 mutant, PsnAP1-2 35S::PsnAP1-2 line in ap1-10 mutant. The arrows represent the locations of observation.
Mentions: To further test the function of PsnAP1-1 and PsnAP1-2, both ORF sequences, which were under a control of a 35S promoter, were introduced into Arabidopsis ap1-10 mutant plants. In the Columbia background, ap1-10 is a strong allele that causes sepals to convert to bract-like structures. Analysis of T3 lines showed that the ap1-10 plants overexpressed with the PsnAP1-1 or PsnAP1-2 were morphologically distinguishable from control plants. However, overexpression of PsnAP1 didn’t complement the ap1-10 phenotype (Fig. 11C–E). The 35S::PsnAP1 ap1-10 plants had reduced height, exhibited less branching, and terminated growth prematurely, producing one ap1-10 like flower each (Fig. 11F, K–M). Flowering time and the number of rosette leaves number were significantly less in the transgenic ap1-10 plants than those in non-transgenic ap1-10 plants (Table 1). Under short-day growth conditions, ap1-10 plants overexpressed with the PsnAP1-1 or PsnAP1-2 also flowered after an average of 10 d, but no visible inflorescence shoots were detected on the original ap1-10 mutants (Fig. 11B, G, H). A few transgenic lines also showed fused terminal flower, curled leaves, and solitary flowers (Fig. 11I, G, K–M).

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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