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Sucrose metabolism gene families and their biological functions.

Jiang SY, Chi YH, Wang JZ, Zhou JX, Cheng YS, Zhang BL, Ma A, Vanitha J, Ramachandran S - Sci Rep (2015)

Bottom Line: Although studies on general metabolism pathway were well documented, less information is available on the genome-wide identification of these genes, their expansion and evolutionary history as well as their biological functions.They were evolutionarily conserved under purifying selection among species and expression divergence played important roles for gene survival after expansion.Overexpression of 15 sorghum genes in Arabidopsis revealed their roles in biomass accumulation, flowering time control, seed germination and response to high salinity and sugar stresses.

View Article: PubMed Central - PubMed

Affiliation: Genome Structural Biology Group, Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604.

ABSTRACT
Sucrose, as the main product of photosynthesis, plays crucial roles in plant development. Although studies on general metabolism pathway were well documented, less information is available on the genome-wide identification of these genes, their expansion and evolutionary history as well as their biological functions. We focused on four sucrose metabolism related gene families including sucrose synthase, sucrose phosphate synthase, sucrose phosphate phosphatase and UDP-glucose pyrophosphorylase. These gene families exhibited different expansion and evolutionary history as their host genomes experienced differentiated rates of the whole genome duplication, tandem and segmental duplication, or mobile element mediated gene gain and loss. They were evolutionarily conserved under purifying selection among species and expression divergence played important roles for gene survival after expansion. However, we have detected recent positive selection during intra-species divergence. Overexpression of 15 sorghum genes in Arabidopsis revealed their roles in biomass accumulation, flowering time control, seed germination and response to high salinity and sugar stresses. Our studies uncovered the molecular mechanisms of gene expansion and evolution and also provided new insight into the role of positive selection in intra-species divergence. Overexpression data revealed novel biological functions of these genes in flowering time control and seed germination under normal and stress conditions.

No MeSH data available.


Related in: MedlinePlus

Molecular and phenotypic characterization of transgenic Arabidopsis plants overexpressing sweet sorghum SPP genes.(A) Comparative expression analysis of 3 sorghum SPP genes between grain (BT × 623) and sweet (Keller) sorghum varieties. YL, YR, S, ML, MR, MP and YP were defined as in Figure 7. YS, young seeds; MS, mature seeds. ACT1, an Arabidopsis gene encoding Actin-1 protein with locus name At2g37620. (B) Expression patterns of 3 SPP sorghum genes under various abiotic stresses and sugar treatments. 1, Control (under normal conditions); 2, 30% PEG 0.5 h; 3, 30% PEG 2 h; 4, 250 mM NaCl 2 h; 5, 250 mM NaCl 8 h; 6, Ice-water at 4 °C 2 h; 7, Ice-water at 4 °C8 h; 8, 5% glucose 2 h; 9, 5% glucose 6 h; 10, 5% sucrose 2 h; 11, 5% sucrose 6 h. (C) Two of three sorghum SPP genes were independently overexpressed in Arabidopsis under the maize ubiquitin promoter to generate at least 4 independent transgenic lines with single copy of T-DNA insertion in each construct. NA, not available. (D) An example of copy number detection by Southern blot hybridization. DNA samples from 19 transgenic plants (ubiquitin::Sobic.004G151800) were digested by PstI for the hybridization using the probe prepared from the HYGROMYCIN gene. (E) Expression analysis of 4 independent transgenic plants (ubiquitin::Sobic.004G151800 and ubiquitin::Sobic.009G040900) with single copy of T-DNA insertion. In (A–E) the prefix “Sobic.” was omitted in each locus name for convenience. (F) Phenotypic observation of transgenic plants under normal and high salinity treatment by comparing with WT plants. SPP1 and SPP2, transgenic plants from ubiquitin::Sobic.004G151800 and ubiquitin::Sobic.009G040900, respectively. (G) Measurement of root length under high salinity stress for transgenic plants overexpressing SPP2. (H) Investigation of germination rates between WT and transgenic plants SPP2 under normal growth conditions. (I) Effects of high salinity treatments on germination rate in transgenic plants SPP2. The stars “*” in (I) indicated significant difference in germination rates between WT and transgenic plants.
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f8: Molecular and phenotypic characterization of transgenic Arabidopsis plants overexpressing sweet sorghum SPP genes.(A) Comparative expression analysis of 3 sorghum SPP genes between grain (BT × 623) and sweet (Keller) sorghum varieties. YL, YR, S, ML, MR, MP and YP were defined as in Figure 7. YS, young seeds; MS, mature seeds. ACT1, an Arabidopsis gene encoding Actin-1 protein with locus name At2g37620. (B) Expression patterns of 3 SPP sorghum genes under various abiotic stresses and sugar treatments. 1, Control (under normal conditions); 2, 30% PEG 0.5 h; 3, 30% PEG 2 h; 4, 250 mM NaCl 2 h; 5, 250 mM NaCl 8 h; 6, Ice-water at 4 °C 2 h; 7, Ice-water at 4 °C8 h; 8, 5% glucose 2 h; 9, 5% glucose 6 h; 10, 5% sucrose 2 h; 11, 5% sucrose 6 h. (C) Two of three sorghum SPP genes were independently overexpressed in Arabidopsis under the maize ubiquitin promoter to generate at least 4 independent transgenic lines with single copy of T-DNA insertion in each construct. NA, not available. (D) An example of copy number detection by Southern blot hybridization. DNA samples from 19 transgenic plants (ubiquitin::Sobic.004G151800) were digested by PstI for the hybridization using the probe prepared from the HYGROMYCIN gene. (E) Expression analysis of 4 independent transgenic plants (ubiquitin::Sobic.004G151800 and ubiquitin::Sobic.009G040900) with single copy of T-DNA insertion. In (A–E) the prefix “Sobic.” was omitted in each locus name for convenience. (F) Phenotypic observation of transgenic plants under normal and high salinity treatment by comparing with WT plants. SPP1 and SPP2, transgenic plants from ubiquitin::Sobic.004G151800 and ubiquitin::Sobic.009G040900, respectively. (G) Measurement of root length under high salinity stress for transgenic plants overexpressing SPP2. (H) Investigation of germination rates between WT and transgenic plants SPP2 under normal growth conditions. (I) Effects of high salinity treatments on germination rate in transgenic plants SPP2. The stars “*” in (I) indicated significant difference in germination rates between WT and transgenic plants.

Mentions: In the sorghum genome, a total of 3 SPP genes have been identified (Table 1). We carried out the comparative expression analysis between grain (BT × 623) and sweet sorghum (Keller) lines by RT-PCR (Fig. 8A,B). The gene Sobic.004G151800 showed the similar expression patterns between these two cultivars among tested tissues. This gene was constitutively expressed in all tested tissues with similar transcript abundance. In contrast, the gene Sobic.009G040900 was not expressed in a few of tested tissues in both BT × 623 and Keller. In BT × 623, the strong expression was detected in young and mature roots as well as stems; no signal was detected in mature seeds and young panicles; the remaining 5 tissues showed relatively less expression level. In Keller, the strongest expression was detected in both young and mature leaves followed by young roots and stems. No expression in young panicles and very faint signal in mature roots and mature panicles were detected. More interestingly, the gene Sobic.009G041000 was tissue-specific and exhibited the expression divergence between grain and sweet sorghum. In BT × 623, this gene was expressed only in young and mature roots; however, it was leaf-specific in keller (Fig. 8A,B). In addition, we further explored whether these three genes were regulated by some stresses in their expression. The gene Sobic.004G151800 showed no difference in its expression level under various stresses including drought (polyethylene glycol, PEG), high salinity (NaCl), cold (Ice-water), glucose and sucrose (Fig. 8C). The gene Sobic.009G040900 was down-regulated by 30% PEG treatment but was up-regulated by both glucose and sucrose. Its expression was not significantly regulated by both high salinity and cold stresses. Interestingly, the gene Sobic.009G041000 was up-regulated by all the tested 5 stresses.


Sucrose metabolism gene families and their biological functions.

Jiang SY, Chi YH, Wang JZ, Zhou JX, Cheng YS, Zhang BL, Ma A, Vanitha J, Ramachandran S - Sci Rep (2015)

Molecular and phenotypic characterization of transgenic Arabidopsis plants overexpressing sweet sorghum SPP genes.(A) Comparative expression analysis of 3 sorghum SPP genes between grain (BT × 623) and sweet (Keller) sorghum varieties. YL, YR, S, ML, MR, MP and YP were defined as in Figure 7. YS, young seeds; MS, mature seeds. ACT1, an Arabidopsis gene encoding Actin-1 protein with locus name At2g37620. (B) Expression patterns of 3 SPP sorghum genes under various abiotic stresses and sugar treatments. 1, Control (under normal conditions); 2, 30% PEG 0.5 h; 3, 30% PEG 2 h; 4, 250 mM NaCl 2 h; 5, 250 mM NaCl 8 h; 6, Ice-water at 4 °C 2 h; 7, Ice-water at 4 °C8 h; 8, 5% glucose 2 h; 9, 5% glucose 6 h; 10, 5% sucrose 2 h; 11, 5% sucrose 6 h. (C) Two of three sorghum SPP genes were independently overexpressed in Arabidopsis under the maize ubiquitin promoter to generate at least 4 independent transgenic lines with single copy of T-DNA insertion in each construct. NA, not available. (D) An example of copy number detection by Southern blot hybridization. DNA samples from 19 transgenic plants (ubiquitin::Sobic.004G151800) were digested by PstI for the hybridization using the probe prepared from the HYGROMYCIN gene. (E) Expression analysis of 4 independent transgenic plants (ubiquitin::Sobic.004G151800 and ubiquitin::Sobic.009G040900) with single copy of T-DNA insertion. In (A–E) the prefix “Sobic.” was omitted in each locus name for convenience. (F) Phenotypic observation of transgenic plants under normal and high salinity treatment by comparing with WT plants. SPP1 and SPP2, transgenic plants from ubiquitin::Sobic.004G151800 and ubiquitin::Sobic.009G040900, respectively. (G) Measurement of root length under high salinity stress for transgenic plants overexpressing SPP2. (H) Investigation of germination rates between WT and transgenic plants SPP2 under normal growth conditions. (I) Effects of high salinity treatments on germination rate in transgenic plants SPP2. The stars “*” in (I) indicated significant difference in germination rates between WT and transgenic plants.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4663468&req=5

f8: Molecular and phenotypic characterization of transgenic Arabidopsis plants overexpressing sweet sorghum SPP genes.(A) Comparative expression analysis of 3 sorghum SPP genes between grain (BT × 623) and sweet (Keller) sorghum varieties. YL, YR, S, ML, MR, MP and YP were defined as in Figure 7. YS, young seeds; MS, mature seeds. ACT1, an Arabidopsis gene encoding Actin-1 protein with locus name At2g37620. (B) Expression patterns of 3 SPP sorghum genes under various abiotic stresses and sugar treatments. 1, Control (under normal conditions); 2, 30% PEG 0.5 h; 3, 30% PEG 2 h; 4, 250 mM NaCl 2 h; 5, 250 mM NaCl 8 h; 6, Ice-water at 4 °C 2 h; 7, Ice-water at 4 °C8 h; 8, 5% glucose 2 h; 9, 5% glucose 6 h; 10, 5% sucrose 2 h; 11, 5% sucrose 6 h. (C) Two of three sorghum SPP genes were independently overexpressed in Arabidopsis under the maize ubiquitin promoter to generate at least 4 independent transgenic lines with single copy of T-DNA insertion in each construct. NA, not available. (D) An example of copy number detection by Southern blot hybridization. DNA samples from 19 transgenic plants (ubiquitin::Sobic.004G151800) were digested by PstI for the hybridization using the probe prepared from the HYGROMYCIN gene. (E) Expression analysis of 4 independent transgenic plants (ubiquitin::Sobic.004G151800 and ubiquitin::Sobic.009G040900) with single copy of T-DNA insertion. In (A–E) the prefix “Sobic.” was omitted in each locus name for convenience. (F) Phenotypic observation of transgenic plants under normal and high salinity treatment by comparing with WT plants. SPP1 and SPP2, transgenic plants from ubiquitin::Sobic.004G151800 and ubiquitin::Sobic.009G040900, respectively. (G) Measurement of root length under high salinity stress for transgenic plants overexpressing SPP2. (H) Investigation of germination rates between WT and transgenic plants SPP2 under normal growth conditions. (I) Effects of high salinity treatments on germination rate in transgenic plants SPP2. The stars “*” in (I) indicated significant difference in germination rates between WT and transgenic plants.
Mentions: In the sorghum genome, a total of 3 SPP genes have been identified (Table 1). We carried out the comparative expression analysis between grain (BT × 623) and sweet sorghum (Keller) lines by RT-PCR (Fig. 8A,B). The gene Sobic.004G151800 showed the similar expression patterns between these two cultivars among tested tissues. This gene was constitutively expressed in all tested tissues with similar transcript abundance. In contrast, the gene Sobic.009G040900 was not expressed in a few of tested tissues in both BT × 623 and Keller. In BT × 623, the strong expression was detected in young and mature roots as well as stems; no signal was detected in mature seeds and young panicles; the remaining 5 tissues showed relatively less expression level. In Keller, the strongest expression was detected in both young and mature leaves followed by young roots and stems. No expression in young panicles and very faint signal in mature roots and mature panicles were detected. More interestingly, the gene Sobic.009G041000 was tissue-specific and exhibited the expression divergence between grain and sweet sorghum. In BT × 623, this gene was expressed only in young and mature roots; however, it was leaf-specific in keller (Fig. 8A,B). In addition, we further explored whether these three genes were regulated by some stresses in their expression. The gene Sobic.004G151800 showed no difference in its expression level under various stresses including drought (polyethylene glycol, PEG), high salinity (NaCl), cold (Ice-water), glucose and sucrose (Fig. 8C). The gene Sobic.009G040900 was down-regulated by 30% PEG treatment but was up-regulated by both glucose and sucrose. Its expression was not significantly regulated by both high salinity and cold stresses. Interestingly, the gene Sobic.009G041000 was up-regulated by all the tested 5 stresses.

Bottom Line: Although studies on general metabolism pathway were well documented, less information is available on the genome-wide identification of these genes, their expansion and evolutionary history as well as their biological functions.They were evolutionarily conserved under purifying selection among species and expression divergence played important roles for gene survival after expansion.Overexpression of 15 sorghum genes in Arabidopsis revealed their roles in biomass accumulation, flowering time control, seed germination and response to high salinity and sugar stresses.

View Article: PubMed Central - PubMed

Affiliation: Genome Structural Biology Group, Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604.

ABSTRACT
Sucrose, as the main product of photosynthesis, plays crucial roles in plant development. Although studies on general metabolism pathway were well documented, less information is available on the genome-wide identification of these genes, their expansion and evolutionary history as well as their biological functions. We focused on four sucrose metabolism related gene families including sucrose synthase, sucrose phosphate synthase, sucrose phosphate phosphatase and UDP-glucose pyrophosphorylase. These gene families exhibited different expansion and evolutionary history as their host genomes experienced differentiated rates of the whole genome duplication, tandem and segmental duplication, or mobile element mediated gene gain and loss. They were evolutionarily conserved under purifying selection among species and expression divergence played important roles for gene survival after expansion. However, we have detected recent positive selection during intra-species divergence. Overexpression of 15 sorghum genes in Arabidopsis revealed their roles in biomass accumulation, flowering time control, seed germination and response to high salinity and sugar stresses. Our studies uncovered the molecular mechanisms of gene expansion and evolution and also provided new insight into the role of positive selection in intra-species divergence. Overexpression data revealed novel biological functions of these genes in flowering time control and seed germination under normal and stress conditions.

No MeSH data available.


Related in: MedlinePlus