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Functional inactivation of UDP-N-acetylglucosamine pyrophosphorylase 1 (UAP1) induces early leaf senescence and defence responses in rice.

Wang Z, Wang Y, Hong X, Hu D, Liu C, Yang J, Li Y, Huang Y, Feng Y, Gong H, Li Y, Fang G, Tang H, Li Y - J. Exp. Bot. (2014)

Bottom Line: The SPL29 gene was identified by map-based cloning, and SPL29 was confirmed as UDP-N-acetylglucosamine pyrophosphorylase 1 (UAP1) by enzymatic analysis.ROS and plant hormones probably play important roles in early leaf senescence and defence responses in the spl29 mutant.Based on these findings, it is suggested that UAP1 is involved in regulating leaf senescence and defence responses in rice.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Hubei 430072, China.

No MeSH data available.


Related in: MedlinePlus

Map-based cloning and functional complementation of SPL29. (A) Preliminary mapping of the SPL29 locus. (B) Fine mapping of the SPL29 locus. (C) Ten putative ORFs are located in the 97-kb region identified by fine mapping. (D) Gene structure of the SPL29 candidate LOC_Os08g10600. Exons, introns, and upstream/downstream regions are displayed. The point mutation of G to T on the eighth exon is indicated by the red arrow, leading to the amino acid exchange of Gly to Cys. (E) Schematic diagram of vectors for functional complementation. The pSPL29C functional complementation vector contains the promoter, gene region, and terminator of LOC_Os08g10600; pEmvC is the empty vector control. The pSPL29C and pEmvC vectors were transformed into spl29 calli. LB, left border; RB, right border; 35S, cauliflower mosaic virus 35S promoter; Bar, the phosphinothricin gene; Tnos, the nopaline synthase terminator. (F–G) Transgenic plants of pSPL29C and pEmvC about 40 days after regeneration. Arrows indicate leaves with the mutant phenotype. (H) Clear leaf phenotype in transgenic plants of pSPL29C and pEmvC. (I) Positive amplification of the transgenic marker element (Bar gene) in transgenic plants of pSPL29C and pEmvC. ZH11 DNA was used as the negative control and the plasmid pEmvC was used as the positive control. Bar178, amplicon of the Bar gene with a length of 178bp. (J) Sequence analysis of the G-to-T mutation site in transgenic plants of pSPL29C and pEmvC.
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Figure 2: Map-based cloning and functional complementation of SPL29. (A) Preliminary mapping of the SPL29 locus. (B) Fine mapping of the SPL29 locus. (C) Ten putative ORFs are located in the 97-kb region identified by fine mapping. (D) Gene structure of the SPL29 candidate LOC_Os08g10600. Exons, introns, and upstream/downstream regions are displayed. The point mutation of G to T on the eighth exon is indicated by the red arrow, leading to the amino acid exchange of Gly to Cys. (E) Schematic diagram of vectors for functional complementation. The pSPL29C functional complementation vector contains the promoter, gene region, and terminator of LOC_Os08g10600; pEmvC is the empty vector control. The pSPL29C and pEmvC vectors were transformed into spl29 calli. LB, left border; RB, right border; 35S, cauliflower mosaic virus 35S promoter; Bar, the phosphinothricin gene; Tnos, the nopaline synthase terminator. (F–G) Transgenic plants of pSPL29C and pEmvC about 40 days after regeneration. Arrows indicate leaves with the mutant phenotype. (H) Clear leaf phenotype in transgenic plants of pSPL29C and pEmvC. (I) Positive amplification of the transgenic marker element (Bar gene) in transgenic plants of pSPL29C and pEmvC. ZH11 DNA was used as the negative control and the plasmid pEmvC was used as the positive control. Bar178, amplicon of the Bar gene with a length of 178bp. (J) Sequence analysis of the G-to-T mutation site in transgenic plants of pSPL29C and pEmvC.

Mentions: The F2 population from the cross between Guangzhan63s and spl29 was used to map the SPL29 gene. Using 44 F2 mutant plants, the SPL29 gene was closely linked with marker M1077, and mapped inside markers M1037 and M1230 on chromosome 8, with an equal genetic distance of 1.4 cM to each (Fig. 2A). For fine mapping of the SPL29 gene, more than 3000 F2 individuals were generated and new markers between M1037 and M1230 were designed according to sequence differences between indica and japonica rice. Nine markers showing polymorphisms between Guangzhan63s and spl29 (Supplementary Table S1) were used to screen the recombinants. Using 870 F2 mutant plants, the SPL29 gene was eventually limited to a 97-kb region between the new markers S8 and S26, with one recombinant for each marker (Fig. 2B). Ten putative open reading frames (ORFs) were predicted according to the RGAP website (Fig. 2C). All ten genes were amplified and sequenced. By comparing gene sequences between wild-type and spl29 plants, one point mutation was identified on the fifth gene, LOC_Os08g10600. No DNA sequence changes were found in the other nine genes or the putative promoter region (about 2.2kb) of LOC_Os08g10600. The single nucleotide substitution of guanine (G) to thymine (T) occurred in the eighth exon of LOC_Os08g10600 (at a position of 712bp in the CDS), resulting in a single amino acid change from glycine (Gly) to cysteine (Cys) (Fig. 2D).


Functional inactivation of UDP-N-acetylglucosamine pyrophosphorylase 1 (UAP1) induces early leaf senescence and defence responses in rice.

Wang Z, Wang Y, Hong X, Hu D, Liu C, Yang J, Li Y, Huang Y, Feng Y, Gong H, Li Y, Fang G, Tang H, Li Y - J. Exp. Bot. (2014)

Map-based cloning and functional complementation of SPL29. (A) Preliminary mapping of the SPL29 locus. (B) Fine mapping of the SPL29 locus. (C) Ten putative ORFs are located in the 97-kb region identified by fine mapping. (D) Gene structure of the SPL29 candidate LOC_Os08g10600. Exons, introns, and upstream/downstream regions are displayed. The point mutation of G to T on the eighth exon is indicated by the red arrow, leading to the amino acid exchange of Gly to Cys. (E) Schematic diagram of vectors for functional complementation. The pSPL29C functional complementation vector contains the promoter, gene region, and terminator of LOC_Os08g10600; pEmvC is the empty vector control. The pSPL29C and pEmvC vectors were transformed into spl29 calli. LB, left border; RB, right border; 35S, cauliflower mosaic virus 35S promoter; Bar, the phosphinothricin gene; Tnos, the nopaline synthase terminator. (F–G) Transgenic plants of pSPL29C and pEmvC about 40 days after regeneration. Arrows indicate leaves with the mutant phenotype. (H) Clear leaf phenotype in transgenic plants of pSPL29C and pEmvC. (I) Positive amplification of the transgenic marker element (Bar gene) in transgenic plants of pSPL29C and pEmvC. ZH11 DNA was used as the negative control and the plasmid pEmvC was used as the positive control. Bar178, amplicon of the Bar gene with a length of 178bp. (J) Sequence analysis of the G-to-T mutation site in transgenic plants of pSPL29C and pEmvC.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4321554&req=5

Figure 2: Map-based cloning and functional complementation of SPL29. (A) Preliminary mapping of the SPL29 locus. (B) Fine mapping of the SPL29 locus. (C) Ten putative ORFs are located in the 97-kb region identified by fine mapping. (D) Gene structure of the SPL29 candidate LOC_Os08g10600. Exons, introns, and upstream/downstream regions are displayed. The point mutation of G to T on the eighth exon is indicated by the red arrow, leading to the amino acid exchange of Gly to Cys. (E) Schematic diagram of vectors for functional complementation. The pSPL29C functional complementation vector contains the promoter, gene region, and terminator of LOC_Os08g10600; pEmvC is the empty vector control. The pSPL29C and pEmvC vectors were transformed into spl29 calli. LB, left border; RB, right border; 35S, cauliflower mosaic virus 35S promoter; Bar, the phosphinothricin gene; Tnos, the nopaline synthase terminator. (F–G) Transgenic plants of pSPL29C and pEmvC about 40 days after regeneration. Arrows indicate leaves with the mutant phenotype. (H) Clear leaf phenotype in transgenic plants of pSPL29C and pEmvC. (I) Positive amplification of the transgenic marker element (Bar gene) in transgenic plants of pSPL29C and pEmvC. ZH11 DNA was used as the negative control and the plasmid pEmvC was used as the positive control. Bar178, amplicon of the Bar gene with a length of 178bp. (J) Sequence analysis of the G-to-T mutation site in transgenic plants of pSPL29C and pEmvC.
Mentions: The F2 population from the cross between Guangzhan63s and spl29 was used to map the SPL29 gene. Using 44 F2 mutant plants, the SPL29 gene was closely linked with marker M1077, and mapped inside markers M1037 and M1230 on chromosome 8, with an equal genetic distance of 1.4 cM to each (Fig. 2A). For fine mapping of the SPL29 gene, more than 3000 F2 individuals were generated and new markers between M1037 and M1230 were designed according to sequence differences between indica and japonica rice. Nine markers showing polymorphisms between Guangzhan63s and spl29 (Supplementary Table S1) were used to screen the recombinants. Using 870 F2 mutant plants, the SPL29 gene was eventually limited to a 97-kb region between the new markers S8 and S26, with one recombinant for each marker (Fig. 2B). Ten putative open reading frames (ORFs) were predicted according to the RGAP website (Fig. 2C). All ten genes were amplified and sequenced. By comparing gene sequences between wild-type and spl29 plants, one point mutation was identified on the fifth gene, LOC_Os08g10600. No DNA sequence changes were found in the other nine genes or the putative promoter region (about 2.2kb) of LOC_Os08g10600. The single nucleotide substitution of guanine (G) to thymine (T) occurred in the eighth exon of LOC_Os08g10600 (at a position of 712bp in the CDS), resulting in a single amino acid change from glycine (Gly) to cysteine (Cys) (Fig. 2D).

Bottom Line: The SPL29 gene was identified by map-based cloning, and SPL29 was confirmed as UDP-N-acetylglucosamine pyrophosphorylase 1 (UAP1) by enzymatic analysis.ROS and plant hormones probably play important roles in early leaf senescence and defence responses in the spl29 mutant.Based on these findings, it is suggested that UAP1 is involved in regulating leaf senescence and defence responses in rice.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Hubei 430072, China.

No MeSH data available.


Related in: MedlinePlus