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The nucleolar GTPase nucleostemin-like 1 plays a role in plant growth and senescence by modulating ribosome biogenesis.

Jeon Y, Park YJ, Cho HK, Jung HJ, Ahn TK, Kang H, Pai HS - J. Exp. Bot. (2015)

Bottom Line: Depletion of NSN1 delayed 25S rRNA maturation and biogenesis of the 60S ribosome subunit, and repressed global translation.NSN1-deficient plants exhibited premature leaf senescence, excessive accumulation of reactive oxygen species, and senescence-related gene expression.Taken together, these results suggest that NSN1 plays a crucial role in plant growth and senescence by modulating ribosome biogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Systems Biology, Yonsei University, Seoul 120-749, Korea.

No MeSH data available.


Related in: MedlinePlus

Silencing of NSN1 using VIGS and DEX-inducible RNAi in N. benthamiana and Arabidopsis. (A) Schematic of N. benthamiana NSN1 (NbNSN1) structure and three VIGS constructs (F, N, and C) that contain different NbNSN1 cDNA fragments, as indicated by the bars. (B) Phenotypes of VIGS plants. NbNSN1 VIGS resulted in growth retardation and premature senescence 20 days after infiltration (DAI), as compared with the control TRV. (C) Quantification of total chlorophyll, chlorophyll a, and chlorophyll b contents in TRV and TRV:NbNSN1 plants (20 DAI). The fourth leaf above the infiltrated leaf was used for the analysis. (D) Real-time quantitative RT–PCR analysis of NbNSN1 transcript levels in TRV:NbNSN1(N), TRV:NbNSN1(C), and TRV:NbNSN1(F) plants (14 DAI) using NSN1-A, NSN1-B, and NSN1-C primers, respectively. The fourth leaf above the infiltrated leaf was used for the analysis. The α-tubulin mRNA level was used as control. Data represent the mean ±SD of three replicates per experiment; *P≤0.05; **P≤0.01. (E) Growth retardation and premature senescence in Arabidopsis DEX-inducible NSN1 RNAi lines (#4 and #17) in response to DEX treatment. Seedlings were grown for 18 d on media that contained either ethanol (–DEX) or 20 μM DEX (+DEX). (F) Real-time quantitative RT–PCR analysis of NSN1 transcript levels in the RNAi lines (#4 and #17) grown for 2 weeks on (–)DEX or (+)DEX media. RNA was isolated from the whole seedlings. The UBC10 mRNA level was used as control. (This figure is available in colour at JXB online.)
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Figure 1: Silencing of NSN1 using VIGS and DEX-inducible RNAi in N. benthamiana and Arabidopsis. (A) Schematic of N. benthamiana NSN1 (NbNSN1) structure and three VIGS constructs (F, N, and C) that contain different NbNSN1 cDNA fragments, as indicated by the bars. (B) Phenotypes of VIGS plants. NbNSN1 VIGS resulted in growth retardation and premature senescence 20 days after infiltration (DAI), as compared with the control TRV. (C) Quantification of total chlorophyll, chlorophyll a, and chlorophyll b contents in TRV and TRV:NbNSN1 plants (20 DAI). The fourth leaf above the infiltrated leaf was used for the analysis. (D) Real-time quantitative RT–PCR analysis of NbNSN1 transcript levels in TRV:NbNSN1(N), TRV:NbNSN1(C), and TRV:NbNSN1(F) plants (14 DAI) using NSN1-A, NSN1-B, and NSN1-C primers, respectively. The fourth leaf above the infiltrated leaf was used for the analysis. The α-tubulin mRNA level was used as control. Data represent the mean ±SD of three replicates per experiment; *P≤0.05; **P≤0.01. (E) Growth retardation and premature senescence in Arabidopsis DEX-inducible NSN1 RNAi lines (#4 and #17) in response to DEX treatment. Seedlings were grown for 18 d on media that contained either ethanol (–DEX) or 20 μM DEX (+DEX). (F) Real-time quantitative RT–PCR analysis of NSN1 transcript levels in the RNAi lines (#4 and #17) grown for 2 weeks on (–)DEX or (+)DEX media. RNA was isolated from the whole seedlings. The UBC10 mRNA level was used as control. (This figure is available in colour at JXB online.)

Mentions: Multispecies sequence alignment revealed that plant NSN1 proteins are homologous to yeast Nug1 and human NS, particularly in the N-terminal region and the circularly permutated GTP-binding motifs (Supplementary Fig. S1 available at JXB online). According to the Genevestigator program (https://www.genevestigator.com/), Arabidopsis NSN1 (At3g07050) is constitutively expressed in various tissues, and exhibits high transcript levels throughout plant development (Supplementary Fig. S2A, B). To determine the in vivo effects of NSN1 deficiency in N. benthamiana and A. thaliana, VIGS and DEX-inducible RNAi were performed using the protocol described in Supplementary Methods S1. For VIGS, three different N. benthamiana NSN1 (NbNSN1) cDNA fragments were cloned; these fragments were designated NbNSN1(F), NbNSN1(N), and NbNSN1(C), which contained the 1842bp full-length coding region, a 489bp N-terminal region, and a 638bp C-terminal region of the NbNSN1 cDNA, respectively. The three fragments were cloned into the TRV-based VIGS vector, pTV00, to create TRV:NbNSN1(F), TRV:NbNSN1(N), and TRV:NbNSN1(C) (Fig. 1A). These vectors were transformed into Agrobacterium tumefaciens, and N. benthamiana plants were infiltrated with the Agrobacterium transformants. VIGS using these TRV:NbNSN1 constructs resulted in growth retardation and premature senescence with reduced chlorophyll contents in leaves compared with the TRV control (Fig. 1B, C; Supplementary Fig. S3). Real-time quantitative RT–PCR revealed significantly lower levels of endogenous NbNSN1 transcripts in leaves of TRV:NbNSN1(N), TRV:NbNSN1(C), and TRV:NbNSN1(F) VIGS plants, indicating silencing of NSN1 (Fig. 1D; Supplementary Table S1).


The nucleolar GTPase nucleostemin-like 1 plays a role in plant growth and senescence by modulating ribosome biogenesis.

Jeon Y, Park YJ, Cho HK, Jung HJ, Ahn TK, Kang H, Pai HS - J. Exp. Bot. (2015)

Silencing of NSN1 using VIGS and DEX-inducible RNAi in N. benthamiana and Arabidopsis. (A) Schematic of N. benthamiana NSN1 (NbNSN1) structure and three VIGS constructs (F, N, and C) that contain different NbNSN1 cDNA fragments, as indicated by the bars. (B) Phenotypes of VIGS plants. NbNSN1 VIGS resulted in growth retardation and premature senescence 20 days after infiltration (DAI), as compared with the control TRV. (C) Quantification of total chlorophyll, chlorophyll a, and chlorophyll b contents in TRV and TRV:NbNSN1 plants (20 DAI). The fourth leaf above the infiltrated leaf was used for the analysis. (D) Real-time quantitative RT–PCR analysis of NbNSN1 transcript levels in TRV:NbNSN1(N), TRV:NbNSN1(C), and TRV:NbNSN1(F) plants (14 DAI) using NSN1-A, NSN1-B, and NSN1-C primers, respectively. The fourth leaf above the infiltrated leaf was used for the analysis. The α-tubulin mRNA level was used as control. Data represent the mean ±SD of three replicates per experiment; *P≤0.05; **P≤0.01. (E) Growth retardation and premature senescence in Arabidopsis DEX-inducible NSN1 RNAi lines (#4 and #17) in response to DEX treatment. Seedlings were grown for 18 d on media that contained either ethanol (–DEX) or 20 μM DEX (+DEX). (F) Real-time quantitative RT–PCR analysis of NSN1 transcript levels in the RNAi lines (#4 and #17) grown for 2 weeks on (–)DEX or (+)DEX media. RNA was isolated from the whole seedlings. The UBC10 mRNA level was used as control. (This figure is available in colour at JXB online.)
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Figure 1: Silencing of NSN1 using VIGS and DEX-inducible RNAi in N. benthamiana and Arabidopsis. (A) Schematic of N. benthamiana NSN1 (NbNSN1) structure and three VIGS constructs (F, N, and C) that contain different NbNSN1 cDNA fragments, as indicated by the bars. (B) Phenotypes of VIGS plants. NbNSN1 VIGS resulted in growth retardation and premature senescence 20 days after infiltration (DAI), as compared with the control TRV. (C) Quantification of total chlorophyll, chlorophyll a, and chlorophyll b contents in TRV and TRV:NbNSN1 plants (20 DAI). The fourth leaf above the infiltrated leaf was used for the analysis. (D) Real-time quantitative RT–PCR analysis of NbNSN1 transcript levels in TRV:NbNSN1(N), TRV:NbNSN1(C), and TRV:NbNSN1(F) plants (14 DAI) using NSN1-A, NSN1-B, and NSN1-C primers, respectively. The fourth leaf above the infiltrated leaf was used for the analysis. The α-tubulin mRNA level was used as control. Data represent the mean ±SD of three replicates per experiment; *P≤0.05; **P≤0.01. (E) Growth retardation and premature senescence in Arabidopsis DEX-inducible NSN1 RNAi lines (#4 and #17) in response to DEX treatment. Seedlings were grown for 18 d on media that contained either ethanol (–DEX) or 20 μM DEX (+DEX). (F) Real-time quantitative RT–PCR analysis of NSN1 transcript levels in the RNAi lines (#4 and #17) grown for 2 weeks on (–)DEX or (+)DEX media. RNA was isolated from the whole seedlings. The UBC10 mRNA level was used as control. (This figure is available in colour at JXB online.)
Mentions: Multispecies sequence alignment revealed that plant NSN1 proteins are homologous to yeast Nug1 and human NS, particularly in the N-terminal region and the circularly permutated GTP-binding motifs (Supplementary Fig. S1 available at JXB online). According to the Genevestigator program (https://www.genevestigator.com/), Arabidopsis NSN1 (At3g07050) is constitutively expressed in various tissues, and exhibits high transcript levels throughout plant development (Supplementary Fig. S2A, B). To determine the in vivo effects of NSN1 deficiency in N. benthamiana and A. thaliana, VIGS and DEX-inducible RNAi were performed using the protocol described in Supplementary Methods S1. For VIGS, three different N. benthamiana NSN1 (NbNSN1) cDNA fragments were cloned; these fragments were designated NbNSN1(F), NbNSN1(N), and NbNSN1(C), which contained the 1842bp full-length coding region, a 489bp N-terminal region, and a 638bp C-terminal region of the NbNSN1 cDNA, respectively. The three fragments were cloned into the TRV-based VIGS vector, pTV00, to create TRV:NbNSN1(F), TRV:NbNSN1(N), and TRV:NbNSN1(C) (Fig. 1A). These vectors were transformed into Agrobacterium tumefaciens, and N. benthamiana plants were infiltrated with the Agrobacterium transformants. VIGS using these TRV:NbNSN1 constructs resulted in growth retardation and premature senescence with reduced chlorophyll contents in leaves compared with the TRV control (Fig. 1B, C; Supplementary Fig. S3). Real-time quantitative RT–PCR revealed significantly lower levels of endogenous NbNSN1 transcripts in leaves of TRV:NbNSN1(N), TRV:NbNSN1(C), and TRV:NbNSN1(F) VIGS plants, indicating silencing of NSN1 (Fig. 1D; Supplementary Table S1).

Bottom Line: Depletion of NSN1 delayed 25S rRNA maturation and biogenesis of the 60S ribosome subunit, and repressed global translation.NSN1-deficient plants exhibited premature leaf senescence, excessive accumulation of reactive oxygen species, and senescence-related gene expression.Taken together, these results suggest that NSN1 plays a crucial role in plant growth and senescence by modulating ribosome biogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Systems Biology, Yonsei University, Seoul 120-749, Korea.

No MeSH data available.


Related in: MedlinePlus