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| Cloning and Expression Analysis of Sucrose Transporter Gene PsSUT2 from Tree Peony (Paeonia suffruticosa) |
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Abstract Sucrose which involved in the plant growth and development processes, the signal transduction pathway and other physiological activities is an important carbon source in plants. Sucrose transporter genes (SUT) play a key role in the long distance transport activities of sucrose in plants. In order to understand the transport mechanism of the SUT in Paeonia suffruticosa, P. suffruticosa 'Luoyanghong' was used as experimental materials in this study. The sucrose transporter gene named PsSUT2 was separated and cloned from the leaf by reverse transcription-PCR (RT-PCR) and rapid-amplification of cDNA ends (RACE). The full length cDNA of PsSUT2 is 2 347 bp, and the ORF encoding a protein of 601 amino acids, was 1 803 bp. The accession number is KC542396 in GenBank. Its molecular mass was 64.6 kD and pI was 6.14, The homologous amino acid sequence of PsSUT2 compared with that of SUT gene from Vitis vinifera was 75.27%. PsSUT2 protein contained 12 transmembrane helical structures, which belongs to the major facilitator superfamily (MFS). The relative expression of PsSUT2 gene and the soluble content in different organs were determined with qRT-PCR and chromatography technology at the different developmental stage of tree peony. The main results showed that the PsSUT2 gene expressed abundantly in the petals at the flowering stage, and the relative expression was 11.18, which was significantly different from other organs of the same stage even every organ of other stages. Meanwhile, the PsSUT2 gene expressed in every organ such as the root, the perennial stem, the leaf, the bulbil, the petal at different developmental stage, but the relative expression were low, for example, the relative expression of petals in the flowering stage was 4.26 times as much as that of roots in the same stage, and 4.40 times as much as that of roots in the little bell stage, and 10.48 times as much as that of petals in the peach stage, and 30.55 times as much as that of bulbil in the bulbil differentiation stage. So, it indicated that the PsSUT2 gene took part in the sucrose unloading activities in the sink organs probably. The contents of sucrose, glucose and fructose of every organ were significantly different at different developmental stage of tree peony, and there was no sucrose almost but much more glucose and fructose in the bulbil or petal before the arrival of flowering. So, the petal of P. suffruticosa 'Luoyanghong' accumulated hexose (glucose and fructose) mainly and the accumulation of hexose was benifit to the flower opening. However, the differences were significantly negative between the sucrose content of leaf, the glucose content of root and the relative expression level of PsSUT2 by the correlative analysis method. Therefore, The expression of the PsSUT2 gene may be inhibited by the sucrose in the leaves and the glucose accumulation in the roots. The results provide a theoretical basis for further understanding of the function of the PsSUT2 gene, and the next step, the gene can be used to carry out molecular breeding and genetic improvement work on the tree peony.
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Received: 09 November 2016
Published: 31 March 2017
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| 白雪梅，张立军，吴晓丹，等．植物蔗糖转运蛋白，植物生理学通讯，2006，42（6）： 1195-1202.（Bai X M, Zhang L J,
Wu X D, et al. 2006. Sucrose trnansporters in Plants[J]. Plant Physiology Communications, 42（6）: 1195-1202.）
刘晓娟．2012．根域限制对牡丹碳代谢和内源激素变化的影响[D]．博士学位论文，河南农业大学，导师：杨秋生．pp.59-
62.(Liu X J. 2012. Carbohydrate Metabolism and Endogenous Hormones Changes of Tree Peony (Paeonia suffruticosa ) Grown
under Root Restriction[D]. Thesis for Ph.D., Henan Agricultural University, Supervisor: Yang Q S. pp.59-62.)
李志鹏．2006．玉米蔗糖转运载体（ZmSUT2和ZmSUT4）的基因克隆及其功能鉴定[D]．硕士学位论文，北京师范大学，导师：王英典.pp.48-50.（Li Z P. 2006. Cloning and functional identification of the ZmSUT2 Gene and ZmSUT4 gene of corn [D]. Thesis for M.S. Beijing Normal University, Supervisor: Wang Y D. pp.48-50.）
史国安，郭香凤，张国海，等．2009．牡丹开花和衰老期间花瓣糖代谢的研究[J]．园艺学报，36(8)：1184－1l90．(Shi G A,
Guo X F, Zhang G H, et al. 2009. Analysis of sugar metabolism during florescence and flower senescence of tree peony petal[J],
Acta Horticulturae Sinica, 36(8): 1184-1l90.)
杨彩菊，郝大海，杨素祥，等．2006．高等植物中的蔗糖载体 ，植物生理学通讯，42（4）：767–776．(Yang C J, Hao D H,
Yang S X, et al. 2006. Sucrose transporters in higher plants [J]. Plant Physiology Communications, 42（4）: 767-776.)
郑乾明. 2015. 甜橙糖转运子基因分离及其在果实糖积累中的功能研究[D]．博士学位论文，华中农业大学，导师：邓秀
新．pp.58-61 (Zheng Q M. 2015. Isolation and functional characterization of sugar transporter genes during fruit sugar
accumulation of sweet orange[D]. Thesis for Ph.D., Huazhong Agricultural University, Supervisor: Deng X H. pp.58- 61. )
Barker L, Kühn C, Weise A, et al. 2000. SUT2, A putative sucrose sensor in sieve elements[J]. Plant Cell. 12: 1153-1164.
Barth I, Meyer S, Sauer N. 2003. PmSUC3: characterazation of a SUT2/SUC3-type sucrose transporter from Plantago major[J]. Plant
Cell, 15(6): l375-1 385.
Btlrkle L, Hibberd J M, Quick W P, et al. 1998. The H+-sucrose cotransporter NtSUTl is essential for sugar export from tobacco leave[J].
Plant Physiology, 118: 59-68.
Buchanan B B, Gruissem W, Jones R L., ed. 2000, Biochemistry & molecular biology of plants[M]. American Society of Plant
Physiologists Prees, Rockville, American, pp.748-776.
Bush D R. 1993. Proton coupled sugar and amino acid transporters in plants[J]. Annual review of plant biology, 44(1): 513-542.
Chun Y L, Jian X S, David W, et al. 2003. Sugars regulate sucrose transporter gene expression in citrus[J]. Biochemical and Biophysical
Research Communications, 306: 402-407.
Ding B, Parthasarathy M V, Niklas K, et al. 1988. A morphometric analysis of the phloem-unloading pathway in developing tobacco
leaves[J]. Planta, 176, 307-318.
Endler A, Meyer S, Schelbert S, et al. 2006. Identification of a vacuolar sucrose transporter in barley and Arabidopsis mesophyll cells by
a tonoplast proteomic approach[J]. Plant Physiology, 141(1): 196-207.
Hackel A, Schauer N, Carrafi F, et al. 2006. Sucrose transporter LeSUTl and LeSUT2 inhibition affects tomato fruit development
indifferent ways[J].Plant Journal, 45(2): 180-192.
Katz E, Fon M, Lee Y J, et al. 2007. The citrus fruit proteome: insights into citrus fruit metabolism[J]. Planta, 226: 989-1005.
Krügel U, Kühn C. 2013. Post-translational regulation of sucrose transporters by direct protein-protein Interactions[J]. Frontier in Plant
Science, 4(2): 59-60.
Kühn C. 2003. A comparison of sucrose transporter systems of different plant species[J]. Plant Biology, 5: 215-232.
Kühn C, Franceschi V, Schulz A, et al. 1997. Macromolecular trafficking indicated by localization and turnover of sucrose transporters
in enucleate sieve elements[J]. Science, 275: 1298-1300.
Kühn C, Grof CP. 2010. Sucrose transporters of higher plants [J]. Current Opinion in Plant Biology, 13(3): 287-297.
Kühn C, Haiirezaei M R, Femie A R, et al. 2003. The sucrose transporter StSUT1 localizes to sieve elements in potato tuber phloem and
influences tuber physiology and development [J]. Plant Physiology, 131:102-113.
Lemoine R. 2000. Sucrose transporters in plants:update on function and structure [J]. Biochimica et Biophysica Acta (BBA)-
Biomembranes, 1465(1-2): 246-262.
Lemoine R, Bnrkle L, Barker L, et al. 1999. Identification of a pollen-specific sucrose transporter-like protein NtSUT3 from tobacco [J].
Febs Letter, 454(3): 325-330.
Li Y H, Guo T, Cui Y, et al. 2015. Cloning and expression of the sucrose transporter gene PsSUT1 from tree peony leaf [J].
Genetics and Molecular Research, 14 (4): 12406-12415.
Lin W, Schmitt M R, Hitz W D, et al. 1984. Sugar transport in isolated corn root protoplasts [J]. Plant Physiology, 76(4): 894- 897.
Meyer S, Lauterbach C, Niedermeier M, et al. 2004. Wounding enhances expression of AtSUC3, a sucrose transponer from Arabidopsis
sieve elements and sink tissues [J]. Plant Physiology，134(2): 684-693.
Meyer S, Melzer M, Truernit E, et al. 2000. AtSUC3, a gene encoding a new Arabidopsis sucrose transporter，is expressed in cells
adjacent to the vascular tissue and in a carpel cell layer [J]. Plant Journal for Cell & Molecular Biology, 24(6):869-882.
Oparka K J, Duckett C M, Prior D A M, et al. 1994. Real-time imaging of phloem unloading in the root tip of Arabidopsis [J]. Plant
Journal 6(6): 759-766.
Rolland N, Ferro M, Seigneurin B D, et al. 2003. Proteomics of chloroplast envelope membranes[J]. Photosynthesis research, 78: 205-
230.
Schmalstig J G, Geiger D R. 1985. Phloem unloading in developing leaves of sugar beet [J]. Plant Physiology, 79(1): 237-241.
Schulze W, Weise A, Frommer W B, et al. 2000. Function of the cytosolic N-terminus of sucrose transporter AtSUT2 in substrate affinity
[J]. FEBS Letters, 485: 189-194.
Shakya R, Sturm A. 1998. Characterization of source-and sink-specific sucrose/H+ symporters from carrot [J]. Plant Physiology, 118:
1473-1480.
Weise A, Barker L, Kühn C, et al. 2000. A new subfamily of sucrose transporters, SUT4, with low affinity/high capacity localized in
sieve elements of plants [J] . Plant Cell, 12(8): l 345-l 356.
Williams L E, Lemoine R, Sauer N. 2000. Sugar transporters in higher plants: a diversjty of foles and comlplex regulation [J]. Trends in
Plant Science, 5(7):283-290.
Yamada K, Ito M, Oyama T. 2007. Analysis of sucrose metabolism during petal growth of cut roses[J]. Postharvest Biology &
Technology, 43(1):174-177.
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