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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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Alignment of the deduced amino acid sequences of AP1 homologs including, PsnAP1-1 and PsnAP1-2.PtrAP1-1 and PtrAP1-2 from Populus. trichocarpa (AAT39554, AAT39556), SdAP1-1 and SdAP1-2 from Salix discolor (AAY82244, AAY82245), PpeAP1 from Prunus persica (ABU63953), PpAP1 from Pyrus pyrifolia (ABP93402), MdAP1 from Malus domestica (BAH10867), EjAP1 from Eriobotrya japonica (AAX14151), MiAP1-1 and MiAP1-2 from Mangifera indica (ACL68407, ACL68408), CsAP1 from Citrus sinensis (AAR01227), RhAP1 from Rosa hybrid cultivar (ACS74806), VvAP1 from Vitis vinifera (ACZ26529), LjAP1 from Lotus japonicus (AAX13296), StAP1-1 and StAP1-2 from Solanum tuberosum (XP_006345101, NP_001275142), NtAP1 from Nicotiana tabacum (AAD01421), ZmAP1-1 and ZmAP1-2 from Zea mays (DAA59399, DAA42679), BoAP1-1 and BoAP1-2 from Brassica oleracea (CAD47853, AAB08875), and AtAP1 from Arabidopsis thaliana (NP_177074). Black shadows indicate identical amino acids; dashed lines indicate gaps to optimize the alignment. The same motif in C terminus is in red square.
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pone-0111725-g001: Alignment of the deduced amino acid sequences of AP1 homologs including, PsnAP1-1 and PsnAP1-2.PtrAP1-1 and PtrAP1-2 from Populus. trichocarpa (AAT39554, AAT39556), SdAP1-1 and SdAP1-2 from Salix discolor (AAY82244, AAY82245), PpeAP1 from Prunus persica (ABU63953), PpAP1 from Pyrus pyrifolia (ABP93402), MdAP1 from Malus domestica (BAH10867), EjAP1 from Eriobotrya japonica (AAX14151), MiAP1-1 and MiAP1-2 from Mangifera indica (ACL68407, ACL68408), CsAP1 from Citrus sinensis (AAR01227), RhAP1 from Rosa hybrid cultivar (ACS74806), VvAP1 from Vitis vinifera (ACZ26529), LjAP1 from Lotus japonicus (AAX13296), StAP1-1 and StAP1-2 from Solanum tuberosum (XP_006345101, NP_001275142), NtAP1 from Nicotiana tabacum (AAD01421), ZmAP1-1 and ZmAP1-2 from Zea mays (DAA59399, DAA42679), BoAP1-1 and BoAP1-2 from Brassica oleracea (CAD47853, AAB08875), and AtAP1 from Arabidopsis thaliana (NP_177074). Black shadows indicate identical amino acids; dashed lines indicate gaps to optimize the alignment. The same motif in C terminus is in red square.

Mentions: Two homologous AP1 cDNAs were cloned from poplar male flower buds of P. simonii × P. nigra using RT-PCR, and named PsnAP1-1 (GenBank No. KC866354) and PsnAP1-2 (GenBank No. KC866355). PsnAP1-1 contains a 726 bp open reading frame (ORF) corresponding to a deduced protein of 241 amino acids, and the estimated molecular weight and isoelectric point of the putative protein were 28.1 kD and 8.19. PsnAP1-2 contains an open reading frame (ORF) of 750 bp, encoding 249 amino acids with a predicted molecular mass of 28.7 kD and a pI of 9.07. The two protein sequences exhibited significant Pfam matches with SRF-type transcription factor (9–59 aa; PF00319) and K-box region (78–174 aa; PF01486) (http://pfam.sanger.ac.uk/), suggesting that these two genes belong to the MADS gene family. PsnAP1-1 shared 82% identity with PsnAP1-2 at the amino acid level. PsnAP1-1 and PsnAP1-2 shared 71% and 67% homology with A. thaliana AP1, respectively (Fig. 1), indicating similar functions of these three proteins. The deduced protein sequences of PsnAP1 also shared high sequence homology with the AP1 proteins previously characterized in other plants (Fig. 1). The two PsnAP1 genes are located in two distinct chromosomes: PsnAP1-1 gene on chromosome 8 and PsnAP1-2 gene on chromosome 10, indicating that they are paralogs instead of alleles.


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)

Alignment of the deduced amino acid sequences of AP1 homologs including, PsnAP1-1 and PsnAP1-2.PtrAP1-1 and PtrAP1-2 from Populus. trichocarpa (AAT39554, AAT39556), SdAP1-1 and SdAP1-2 from Salix discolor (AAY82244, AAY82245), PpeAP1 from Prunus persica (ABU63953), PpAP1 from Pyrus pyrifolia (ABP93402), MdAP1 from Malus domestica (BAH10867), EjAP1 from Eriobotrya japonica (AAX14151), MiAP1-1 and MiAP1-2 from Mangifera indica (ACL68407, ACL68408), CsAP1 from Citrus sinensis (AAR01227), RhAP1 from Rosa hybrid cultivar (ACS74806), VvAP1 from Vitis vinifera (ACZ26529), LjAP1 from Lotus japonicus (AAX13296), StAP1-1 and StAP1-2 from Solanum tuberosum (XP_006345101, NP_001275142), NtAP1 from Nicotiana tabacum (AAD01421), ZmAP1-1 and ZmAP1-2 from Zea mays (DAA59399, DAA42679), BoAP1-1 and BoAP1-2 from Brassica oleracea (CAD47853, AAB08875), and AtAP1 from Arabidopsis thaliana (NP_177074). Black shadows indicate identical amino acids; dashed lines indicate gaps to optimize the alignment. The same motif in C terminus is in red square.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4216142&req=5

pone-0111725-g001: Alignment of the deduced amino acid sequences of AP1 homologs including, PsnAP1-1 and PsnAP1-2.PtrAP1-1 and PtrAP1-2 from Populus. trichocarpa (AAT39554, AAT39556), SdAP1-1 and SdAP1-2 from Salix discolor (AAY82244, AAY82245), PpeAP1 from Prunus persica (ABU63953), PpAP1 from Pyrus pyrifolia (ABP93402), MdAP1 from Malus domestica (BAH10867), EjAP1 from Eriobotrya japonica (AAX14151), MiAP1-1 and MiAP1-2 from Mangifera indica (ACL68407, ACL68408), CsAP1 from Citrus sinensis (AAR01227), RhAP1 from Rosa hybrid cultivar (ACS74806), VvAP1 from Vitis vinifera (ACZ26529), LjAP1 from Lotus japonicus (AAX13296), StAP1-1 and StAP1-2 from Solanum tuberosum (XP_006345101, NP_001275142), NtAP1 from Nicotiana tabacum (AAD01421), ZmAP1-1 and ZmAP1-2 from Zea mays (DAA59399, DAA42679), BoAP1-1 and BoAP1-2 from Brassica oleracea (CAD47853, AAB08875), and AtAP1 from Arabidopsis thaliana (NP_177074). Black shadows indicate identical amino acids; dashed lines indicate gaps to optimize the alignment. The same motif in C terminus is in red square.
Mentions: Two homologous AP1 cDNAs were cloned from poplar male flower buds of P. simonii × P. nigra using RT-PCR, and named PsnAP1-1 (GenBank No. KC866354) and PsnAP1-2 (GenBank No. KC866355). PsnAP1-1 contains a 726 bp open reading frame (ORF) corresponding to a deduced protein of 241 amino acids, and the estimated molecular weight and isoelectric point of the putative protein were 28.1 kD and 8.19. PsnAP1-2 contains an open reading frame (ORF) of 750 bp, encoding 249 amino acids with a predicted molecular mass of 28.7 kD and a pI of 9.07. The two protein sequences exhibited significant Pfam matches with SRF-type transcription factor (9–59 aa; PF00319) and K-box region (78–174 aa; PF01486) (http://pfam.sanger.ac.uk/), suggesting that these two genes belong to the MADS gene family. PsnAP1-1 shared 82% identity with PsnAP1-2 at the amino acid level. PsnAP1-1 and PsnAP1-2 shared 71% and 67% homology with A. thaliana AP1, respectively (Fig. 1), indicating similar functions of these three proteins. The deduced protein sequences of PsnAP1 also shared high sequence homology with the AP1 proteins previously characterized in other plants (Fig. 1). The two PsnAP1 genes are located in two distinct chromosomes: PsnAP1-1 gene on chromosome 8 and PsnAP1-2 gene on chromosome 10, indicating that they are paralogs instead of alleles.

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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