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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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Northern blot analysis of PsnAP1-1 and PsnAP1-2 expression in transgenic plants.(A, B) Schematic of the T-DNA region of the binary vector pROKII-PsnAP1-1 and pROKII-PsnAP1-2. LB left border, Pnos nopalina synthase promoter, NptII, kanamycin resistance gene, Tnos nopalina synthase terminator, P35S CaMV 35S promoter, RB right border. (C) Northern blot analysis of 35S::PsnAP1-1 in transgenic tobacco. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-1 lines. (D) Northern blot analysis of 35S::PsnAP1-2 in transgenic tobacco. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L6–10 five 35S::PsnAP1-2 lines. (E) Northern blot analysis of 35S::PsnAP1-1 in transgenic Arabidopsis. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-1 lines, ap1-10 ap1-10 mutant, T1–3 three 35S::PsnAP1-1 lines in ap1-10 mutants. (F) Northern blot analysis of 35S::PsnAP1-2 in transgenic Arabidopsis. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-2 lines, ap1-10 ap1-10 mutant, T1–3 three 35S::PsnAP1-2 lines in ap1-10 mutants.
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pone-0111725-g004: Northern blot analysis of PsnAP1-1 and PsnAP1-2 expression in transgenic plants.(A, B) Schematic of the T-DNA region of the binary vector pROKII-PsnAP1-1 and pROKII-PsnAP1-2. LB left border, Pnos nopalina synthase promoter, NptII, kanamycin resistance gene, Tnos nopalina synthase terminator, P35S CaMV 35S promoter, RB right border. (C) Northern blot analysis of 35S::PsnAP1-1 in transgenic tobacco. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-1 lines. (D) Northern blot analysis of 35S::PsnAP1-2 in transgenic tobacco. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L6–10 five 35S::PsnAP1-2 lines. (E) Northern blot analysis of 35S::PsnAP1-1 in transgenic Arabidopsis. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-1 lines, ap1-10 ap1-10 mutant, T1–3 three 35S::PsnAP1-1 lines in ap1-10 mutants. (F) Northern blot analysis of 35S::PsnAP1-2 in transgenic Arabidopsis. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-2 lines, ap1-10 ap1-10 mutant, T1–3 three 35S::PsnAP1-2 lines in ap1-10 mutants.

Mentions: Northern blotting and qRT-PCR were used to reveal PsnAP1-1 and PsnAP1-2 transcripts in different poplar tissues (Fig. 3). Northern blotting demonstrated that PsnAP1-1 and PsnAP1-2 are highly expressed in both male and female inflorescence but are expressed extremely in low levels in shoots, stems, leaves, and roots (Fig. 3A, B). This expression profile was similar to that of the AP1 gene in birch as reported by Qu et al. [10]. qRT-PCR analysis was used to detect the expression level of PsnAP1 in developmental male floral buds from September to the following April, and results indicated high transcription levels of PsnAP1-1 and PsnAP1-2 from September to November, after which a declining trend took place from December to April (Fig. 4C). This analysis revealed that the transcription of PsnAP1 remained continuous and stable during the floral primordia formation and floral organ differentiation periods, but the transcription of PsnAP1s declined in the inflorescence meristem growth period.


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)

Northern blot analysis of PsnAP1-1 and PsnAP1-2 expression in transgenic plants.(A, B) Schematic of the T-DNA region of the binary vector pROKII-PsnAP1-1 and pROKII-PsnAP1-2. LB left border, Pnos nopalina synthase promoter, NptII, kanamycin resistance gene, Tnos nopalina synthase terminator, P35S CaMV 35S promoter, RB right border. (C) Northern blot analysis of 35S::PsnAP1-1 in transgenic tobacco. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-1 lines. (D) Northern blot analysis of 35S::PsnAP1-2 in transgenic tobacco. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L6–10 five 35S::PsnAP1-2 lines. (E) Northern blot analysis of 35S::PsnAP1-1 in transgenic Arabidopsis. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-1 lines, ap1-10 ap1-10 mutant, T1–3 three 35S::PsnAP1-1 lines in ap1-10 mutants. (F) Northern blot analysis of 35S::PsnAP1-2 in transgenic Arabidopsis. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-2 lines, ap1-10 ap1-10 mutant, T1–3 three 35S::PsnAP1-2 lines in ap1-10 mutants.
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pone-0111725-g004: Northern blot analysis of PsnAP1-1 and PsnAP1-2 expression in transgenic plants.(A, B) Schematic of the T-DNA region of the binary vector pROKII-PsnAP1-1 and pROKII-PsnAP1-2. LB left border, Pnos nopalina synthase promoter, NptII, kanamycin resistance gene, Tnos nopalina synthase terminator, P35S CaMV 35S promoter, RB right border. (C) Northern blot analysis of 35S::PsnAP1-1 in transgenic tobacco. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-1 lines. (D) Northern blot analysis of 35S::PsnAP1-2 in transgenic tobacco. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L6–10 five 35S::PsnAP1-2 lines. (E) Northern blot analysis of 35S::PsnAP1-1 in transgenic Arabidopsis. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-1 lines, ap1-10 ap1-10 mutant, T1–3 three 35S::PsnAP1-1 lines in ap1-10 mutants. (F) Northern blot analysis of 35S::PsnAP1-2 in transgenic Arabidopsis. Leaves were sampled from wild-type and transgenic plants. WT wild-type, L1–5 five 35S::PsnAP1-2 lines, ap1-10 ap1-10 mutant, T1–3 three 35S::PsnAP1-2 lines in ap1-10 mutants.
Mentions: Northern blotting and qRT-PCR were used to reveal PsnAP1-1 and PsnAP1-2 transcripts in different poplar tissues (Fig. 3). Northern blotting demonstrated that PsnAP1-1 and PsnAP1-2 are highly expressed in both male and female inflorescence but are expressed extremely in low levels in shoots, stems, leaves, and roots (Fig. 3A, B). This expression profile was similar to that of the AP1 gene in birch as reported by Qu et al. [10]. qRT-PCR analysis was used to detect the expression level of PsnAP1 in developmental male floral buds from September to the following April, and results indicated high transcription levels of PsnAP1-1 and PsnAP1-2 from September to November, after which a declining trend took place from December to April (Fig. 4C). This analysis revealed that the transcription of PsnAP1 remained continuous and stable during the floral primordia formation and floral organ differentiation periods, but the transcription of PsnAP1s declined in the inflorescence meristem growth period.

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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