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Physalis floridana Cell Number Regulator1 encodes a cell membrane-anchored modulator of cell cycle and negatively controls fruit size.

Li Z, He C - J. Exp. Bot. (2014)

Bottom Line: Physalis species show a significant variation in berry size; however, the underlying molecular basis is unknown.The nuclear import of PfAG2 was essential in the proposed pathway.This study also sheds light on the link between organ identity and organ growth in plants.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, Nanxincun 20, Xiangshan, Haidian, 100093 Beijing, PR China University of Chinese Academy of Sciences, Yuquan Road 19, 100049 Beijing, PR China.

No MeSH data available.


Regulation of the PfCYCD2;1 expression. (A) Gene expression analysis of the putative PfCNR1 interacting partners and the putative Cyclin. Total RNA from the B2 flower buds was subjected to qRT-PCR. PfACTIN was used as an internal control. Expression of each gene was investigated in three independent biological samples and its expression in the wild-type P106 was set as 1. The mean and standard deviation are presented. (B) Characterization of the putative PfCYCD2;1 promoter. Three CArG-boxes (C1, C2, and C3) are highlighted in red circles beyond the ATG. The mutated promoters resulting from either a point mutation (C1pm) or deletion mutation (C1dm) are shown. The putative promoter sequences are presented in Supplementary Fig. S8. (C) Yeast one-hybrid assays between PfAG2 and the related DNA fragments. Pro, promoter; 3×, three tandem repeats of the CArG-box; C1, CArG-box1; C2, CArG-box2; C3, CArG-box3; dm, deleted mutation; pm, point mutation. The survival of yeast cells on SD/–Leumedium supplemented with aureobasidin A (AbA) suggested that the protein could bind to the corresponding DNA fragment. (D) LUC relative activity assay in P. floridana protoplasts. LUC activities were normalized using 35S:GUS as an internal control. The wild-type PfAG2 associated interactions are highlighted in pink and the interactions of PfAG2m that excluded the NLS are indicated in red. The mean and standard deviation from at least six independently repeated assays are presented. The star and the square, respectively, indicate a significant difference (P<0.05) compared with ProPfCYCD2;1:LUC or ProPfCYCD2;1:LUC with PfAG2. (E) The NLS in PfAG2 and its deleted version (PfAG2m). The 25 aa sequences of the NLS are given between the PfAG2 and PfAG2m structures. (F) Subcellular localization of PfAG2. The arrow indicates the nuclei. (G) Subcellular localization of PfAG2m. Bar, 10 μm (F, G).
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Figure 6: Regulation of the PfCYCD2;1 expression. (A) Gene expression analysis of the putative PfCNR1 interacting partners and the putative Cyclin. Total RNA from the B2 flower buds was subjected to qRT-PCR. PfACTIN was used as an internal control. Expression of each gene was investigated in three independent biological samples and its expression in the wild-type P106 was set as 1. The mean and standard deviation are presented. (B) Characterization of the putative PfCYCD2;1 promoter. Three CArG-boxes (C1, C2, and C3) are highlighted in red circles beyond the ATG. The mutated promoters resulting from either a point mutation (C1pm) or deletion mutation (C1dm) are shown. The putative promoter sequences are presented in Supplementary Fig. S8. (C) Yeast one-hybrid assays between PfAG2 and the related DNA fragments. Pro, promoter; 3×, three tandem repeats of the CArG-box; C1, CArG-box1; C2, CArG-box2; C3, CArG-box3; dm, deleted mutation; pm, point mutation. The survival of yeast cells on SD/–Leumedium supplemented with aureobasidin A (AbA) suggested that the protein could bind to the corresponding DNA fragment. (D) LUC relative activity assay in P. floridana protoplasts. LUC activities were normalized using 35S:GUS as an internal control. The wild-type PfAG2 associated interactions are highlighted in pink and the interactions of PfAG2m that excluded the NLS are indicated in red. The mean and standard deviation from at least six independently repeated assays are presented. The star and the square, respectively, indicate a significant difference (P<0.05) compared with ProPfCYCD2;1:LUC or ProPfCYCD2;1:LUC with PfAG2. (E) The NLS in PfAG2 and its deleted version (PfAG2m). The 25 aa sequences of the NLS are given between the PfAG2 and PfAG2m structures. (F) Subcellular localization of PfAG2. The arrow indicates the nuclei. (G) Subcellular localization of PfAG2m. Bar, 10 μm (F, G).

Mentions: Transcriptional regulation by the PfCNR1 signalling pathway were also investigated. The expression of the putative PfCNR1 interacting partner genes was first checked, and they were all expressed but no differential expression in the B2 flower buds was observed among P058, P064, P106, 35S: PfCNR1 (OE9), and 35S: PfCNR1-RNAi (R9) plants (Fig. 6A). Since cell division in eukaryotic organisms is controlled by highly conserved basic cell-cycle machinery (Dewitte and Murray, 2003), we identified five putative Cyclin genes from P. floridana that are key genes in the plant cell division cycle (Mizukami, 2001). We designated these genes PfCYCA2;1, PfCYCB1;1, PfCYCB2;1, PfCYCD2;1, and PfCYCD3;1. In the B2 flower buds of P058, P064, R9, and OE9 plants, only PfCYCD2;1 was significantly elevated in P058, P064, and R9 plants, and it was slightly repressed in OE9 plants relative to the wild-type P106 (Fig. 6A). The expression of CyclinD2;1 positively correlated with cell division (Oh et al., 2008; Talengera et al., 2012). Therefore, PfCNR1 may function as a negative modulator of the cell division cycle by directly or indirectly influencing the expression of PfCYCD2;1.


Physalis floridana Cell Number Regulator1 encodes a cell membrane-anchored modulator of cell cycle and negatively controls fruit size.

Li Z, He C - J. Exp. Bot. (2014)

Regulation of the PfCYCD2;1 expression. (A) Gene expression analysis of the putative PfCNR1 interacting partners and the putative Cyclin. Total RNA from the B2 flower buds was subjected to qRT-PCR. PfACTIN was used as an internal control. Expression of each gene was investigated in three independent biological samples and its expression in the wild-type P106 was set as 1. The mean and standard deviation are presented. (B) Characterization of the putative PfCYCD2;1 promoter. Three CArG-boxes (C1, C2, and C3) are highlighted in red circles beyond the ATG. The mutated promoters resulting from either a point mutation (C1pm) or deletion mutation (C1dm) are shown. The putative promoter sequences are presented in Supplementary Fig. S8. (C) Yeast one-hybrid assays between PfAG2 and the related DNA fragments. Pro, promoter; 3×, three tandem repeats of the CArG-box; C1, CArG-box1; C2, CArG-box2; C3, CArG-box3; dm, deleted mutation; pm, point mutation. The survival of yeast cells on SD/–Leumedium supplemented with aureobasidin A (AbA) suggested that the protein could bind to the corresponding DNA fragment. (D) LUC relative activity assay in P. floridana protoplasts. LUC activities were normalized using 35S:GUS as an internal control. The wild-type PfAG2 associated interactions are highlighted in pink and the interactions of PfAG2m that excluded the NLS are indicated in red. The mean and standard deviation from at least six independently repeated assays are presented. The star and the square, respectively, indicate a significant difference (P<0.05) compared with ProPfCYCD2;1:LUC or ProPfCYCD2;1:LUC with PfAG2. (E) The NLS in PfAG2 and its deleted version (PfAG2m). The 25 aa sequences of the NLS are given between the PfAG2 and PfAG2m structures. (F) Subcellular localization of PfAG2. The arrow indicates the nuclei. (G) Subcellular localization of PfAG2m. Bar, 10 μm (F, G).
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Figure 6: Regulation of the PfCYCD2;1 expression. (A) Gene expression analysis of the putative PfCNR1 interacting partners and the putative Cyclin. Total RNA from the B2 flower buds was subjected to qRT-PCR. PfACTIN was used as an internal control. Expression of each gene was investigated in three independent biological samples and its expression in the wild-type P106 was set as 1. The mean and standard deviation are presented. (B) Characterization of the putative PfCYCD2;1 promoter. Three CArG-boxes (C1, C2, and C3) are highlighted in red circles beyond the ATG. The mutated promoters resulting from either a point mutation (C1pm) or deletion mutation (C1dm) are shown. The putative promoter sequences are presented in Supplementary Fig. S8. (C) Yeast one-hybrid assays between PfAG2 and the related DNA fragments. Pro, promoter; 3×, three tandem repeats of the CArG-box; C1, CArG-box1; C2, CArG-box2; C3, CArG-box3; dm, deleted mutation; pm, point mutation. The survival of yeast cells on SD/–Leumedium supplemented with aureobasidin A (AbA) suggested that the protein could bind to the corresponding DNA fragment. (D) LUC relative activity assay in P. floridana protoplasts. LUC activities were normalized using 35S:GUS as an internal control. The wild-type PfAG2 associated interactions are highlighted in pink and the interactions of PfAG2m that excluded the NLS are indicated in red. The mean and standard deviation from at least six independently repeated assays are presented. The star and the square, respectively, indicate a significant difference (P<0.05) compared with ProPfCYCD2;1:LUC or ProPfCYCD2;1:LUC with PfAG2. (E) The NLS in PfAG2 and its deleted version (PfAG2m). The 25 aa sequences of the NLS are given between the PfAG2 and PfAG2m structures. (F) Subcellular localization of PfAG2. The arrow indicates the nuclei. (G) Subcellular localization of PfAG2m. Bar, 10 μm (F, G).
Mentions: Transcriptional regulation by the PfCNR1 signalling pathway were also investigated. The expression of the putative PfCNR1 interacting partner genes was first checked, and they were all expressed but no differential expression in the B2 flower buds was observed among P058, P064, P106, 35S: PfCNR1 (OE9), and 35S: PfCNR1-RNAi (R9) plants (Fig. 6A). Since cell division in eukaryotic organisms is controlled by highly conserved basic cell-cycle machinery (Dewitte and Murray, 2003), we identified five putative Cyclin genes from P. floridana that are key genes in the plant cell division cycle (Mizukami, 2001). We designated these genes PfCYCA2;1, PfCYCB1;1, PfCYCB2;1, PfCYCD2;1, and PfCYCD3;1. In the B2 flower buds of P058, P064, R9, and OE9 plants, only PfCYCD2;1 was significantly elevated in P058, P064, and R9 plants, and it was slightly repressed in OE9 plants relative to the wild-type P106 (Fig. 6A). The expression of CyclinD2;1 positively correlated with cell division (Oh et al., 2008; Talengera et al., 2012). Therefore, PfCNR1 may function as a negative modulator of the cell division cycle by directly or indirectly influencing the expression of PfCYCD2;1.

Bottom Line: Physalis species show a significant variation in berry size; however, the underlying molecular basis is unknown.The nuclear import of PfAG2 was essential in the proposed pathway.This study also sheds light on the link between organ identity and organ growth in plants.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, Nanxincun 20, Xiangshan, Haidian, 100093 Beijing, PR China University of Chinese Academy of Sciences, Yuquan Road 19, 100049 Beijing, PR China.

No MeSH data available.