Tissue Expression Pattern Analysis of TIPs Genes in Phyllostachys edulis

SUN Hua-yu; LOU Yong-feng; LI Li-chao; ZHAO Han-sheng; GAO Zhi-min

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Tissue Expression Pattern Analysis of TIPs Genes in Phyllostachys edulis

1.
International Center for Bamboo and Rattan, State Forestry Administration Key Laboratory on the Science and Technology of Bamboo and Rattan, Beijing 100102, China

Received Date: 2015-08-04

Abstract

[Objective] To reveal the role of tonoplast intrinsic proteins (TIPs) in bamboo under stress conditions and provide new genetic resource for the breeding of new varieties. [Method] The molecular characteristics and expression profiles of TIPs in moso bamboo (Phyllostachys edulis (Carr.) H. de Lehaie) were conducted. The gene expression in different tissues and those under drought, water and NaCl abiotic stresses were analyzed with real-time quantitative PCR. [Result] The result indicated that there were six TIPs homologous genes belonged to three subgroups (TIP1, TIP2 and TIP4) in moso bamboo genome, among which four genes (PeTIP1;1, PeTIP1;2, PeTIP2;2 and PeTIP4;2) consisted of two exons and one intron, and the other two genes (PeTIP2;1 and PeTIP4;1) consisted of three exons and two introns, respectively. Protein structure analysis indicated that all the six PeTIPs had two typical NPA domains and four conserved ar/R selectivity filter. Tissue specific analysis based on transcriptome demonstrated that PeTIP1;1 expressed with high levels in all tissues, PeTIP1;2 had the highest expression level in flower, PeTIP2;1 was mainly expressed in root and rhizome, PeTIP2;2 was specifically expressed in roots, PeTIP4;1 had the highest expression level in leaf, and PeTIP4;2 expressed in shoot and rhizome with high level, but the lowest in the roots. The result of qRT-PCR confirmed that in roots PeTIP4;1 was up-regulated, while PeTIP2;1, PeTIP2;2 and PeTIP4;2 were inhibited significantly (pPeTIP1;1 and PeTIP4;1 increased and those of PeTIP1;2, PeTIP2;2, and PeTIP4;2 decreased significantly (pPeTIPs all raised significantly (pPeTIP1;1, PeTIP1;2 and PeTIP4;1 were up-regulated under drought treatment, all the expression levels of six PeTIPs were significantly increased under water stress (pPeTIP2;1 was suppressed significantly (p[Conclusion] This result indicated that PeTIPs may play roles with varying degrees in bamboo tolerance of drought, water, NaCl and other abiotic stresses.
- Phyllostachys edulis,
- tonoplast intrinsic proteins,
- expression analysis

References

[1]	Mueller N D, Gerber J S, Johnston M, et al. Closing yield gaps through nutrient and water management[J], Nature, 2012,490(7419):254-257.
[2]	Kaldenhoff R, Fischer M. Functional aquaporin diversity in plants[J]. Biochimica et Biophysica Acta, 2006, 1758(8):1134-1141.
[3]	Henzler T, Steudle E. Reversible closing of water channels in Chara internodes provides evidence for a composite transport model of the plasma membrane[J]. Journal of Experimental Botany, 1995, 46(2):199-209.
[4]	Chen S, Polle A. Salinity tolerance of Populus[J]. Plant Biology, 2010,12(2):317-333.
[5]	Ruiz-Lozano J M, Porcel R, Azcón C, et al. Regulation by arbuscular mycorrhizae of the integrated physiological response to salinity in plants: new challenges in physiological and molecular studies[J]. Journal of Experimental Botany, 2012, 63(11):4033-4044.
[6]	Maurel C, Tacnet F, Guclu J, et al. Purified vesicles of tobacco cell vacuolar and plasma membranes exhibit dramatically different water permeability and water channel activity[J]. Proceedings of the National Academy of Sciences of the United States of America, 1997, 94(13):7103-7108.
[7]	Regon P, Panda P, Kshetrimayum E, et al. Genome-wide comparative analysis of tonoplast intrinsic protein (TIP) genes in plants[J]. Functional & Integrative Genomics, 2014, 14(4):617-629.
[8]	Sade N, Vinocur B J, Diber A, et al. Improving plant stress tolerance and yield production: is the tonoplast aquaporin SlTIP2;2 a key to isohydric to anisohydric conversion? [J]. New Phytologist, 2009, 181(3):651-661.
[9]	Wang L L, Chen A P, Zhong N Q, et al. The Thellungiella salsuginea tonoplast aquaporin TsTIP1;2 functions in protection against multiple abiotic stresses[J]. Plant Cell Physiology, 2014, 55(1):148-161.
[10]	Peng Y, Lin W, Cai W, et al. Overexpression of a Panax ginseng tonoplast aquaporin alters salt tolerance, drought tolerance and cold acclimation ability in transgenic Arabidopsis plants[J]. Planta, 2007, 226(3):729-740.
[11]	江泽慧.世界竹藤[M].辽宁科学技术出版社,2002,3-10.
[12]	Cui K, He C, Zhang J. Temporal and spatial profiling of internode elongation-associated protein expression in rapidly growing culms of bamboo[J]. Journal of Proteome Research, 2012, 11(4):2492-2507.
[13]	Hepler P K. Calcium: A central regulator of plant growth and development[J]. The Plant Cell, 2005, 17(8):2142-2155.
[14]	Cao K F, Yang S J, Zhang Y J, et al. The maximum height of grasses is determined by roots[J]. Ecology Letters, 2012, 15(7):666-672.
[15]	Nguyen M X, Moon S, Jung K H. Genome-wide expression analysis of rice aquaporin genes and development of a functional gene network mediated by aquaporin expression in roots[J]. Planta, 2013, 238(4):669-681.
[16]	Yue X, Zhao X, Fei Y, et al. Correlation of aquaporins and transmembrane solute transporters revealed by genome-wide analysis in developing maize leaf[J]. Comparative and Functional Genomics, 2012, 2012:546930.
[17]	Zhao H, Peng Z, Fei B, et al. BambooGDB: a bamboo genome database with functional annotation and an analysis platform[J]. Database, 2014, 2014:bau006.
[18]	Gao Z M, Li X P, Li L B, et al. An effective method for total RNA isolation from bamboo[J]. Chinese Forestry Science and Technology, 2006, 5(3):52-54.
[19]	Fan C, Ma J, Guo Q, et al. Selection of reference genes for quantitative real-time PCR in bamboo (Phyllostachys edulis) [J]. PLoS One, 2013, 8(2):e56573.
[20]	Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCt method[J]. Methods, 2001, 25(4):402-408.
[21]	Peng Z, Lu T, Li L, et al. Genome-wide characterization of the biggest grass, bamboo, based on 10,608 putative full-length cDNA sequences[J]. BMC Plant Biology, 2010, 10:116.
[22]	Moore M J, Query C C, Sharp P A. Splicing of precursors to mRNA by the spliceosome[M]. Cold Spring Harbor: Cold Spring Harbor laboratory Press, 1993,303-357.
[23]	Chou K C, Shen H B. Plant-mPLoc: a top-down strategy to augment the power for predicting plant protein subcellular localization[J]. PLoS One, 2010, 5:e11335.
[24]	Wree D, Wu B, Zeuthen T, et al. Requirement for asparagine in the aquaporin NPA sequence signature motifs for cation exclusion[J]. The FEBS Journal, 2011, 278(5):740-748.
[25]	Mitani-Ueno N, Yamaji N, Zhao F J, et al. The aromatic/arginine selectivity filter of NIP aquaporins plays a critical role in substrate selectivity for silicon, boron, and arsenic[J]. Journal of Experimental Botany, 2011, 62(12):4391-4398.
[26]	Reuscher S, Akiyama M, Mori C, et al. Genome-wide identification and expression analysis of aquaporins in tomato[J]. PLoS One, 2013, 8(11):e79052.
[27]	Utsugi S, Shibasaka M, Maekawa M, et al. Control of the water transport activity of barley HvTIP3;1 specifically expressed in seeds[J]. Plant Cell Physiology, 2015, pii:pcv104.
[28]	Forrest K L, Bhave M. The PIP and TIP aquaporins in wheat form a large and diverse family with unique gene structures and functionally important features[J]. Functional & Integrative Genomics, 2008, 8(2):115-133.
[29]	Hove R M, Ziemann M, Bhave M. Identification and expression analysis of the barley (Hordeum vulgare L.) aquaporin gene family[J]. PloS One, 2015, 10(6):e0128025.
[30]	Peng Z H, Lu Y, Li L B, et al. The draft genome of the fast growing non-timber forest species moso bamboo (Phyllostachys heterocycla)[J]. Nature Genetics, 2013, 45(4):456-461.
[31]	Verma R K, Prabh N D, Sankararamakrishnan R. Intra-helical salt-bridge and helix destabilizing residues within the same helical turn: Role of functionally important loop E half-helix in channel regulation of major intrinsic proteins[J]. Biochimica et Biophysica Acta, 2015, 1848(6):1436-1449.
[32]	Zhang D Y, Ali Z, Wang C B, et al. Genome-wide sequence characterization and expression analysis of major intrinsic proteins in soybean (Glycine max L.)[J]. PLoS One, 2013, 8(2):e56312.
[33]	Katsuhara M, Hanba Y T, Shiratake K, et al. Expanding roles of plant aquaporins in plasma membranes and cell organelles[J]. Functional Plant Biology, 2008, 35(1):1-14.
[34]	Schmid M, Davison T S, Henz S R, et al. A gene expression map of Arabidopsis thaliana development[J]. Nature Genetics, 2005, 37(5):501-506.
[35]	Beebo A, Thomas D, Der C, et al. Life with and without AtTIP1;1, an Arabidopsis aquaporin preferentially localized in the apposing tonoplasts of adjacent vacuoles[J]. Plant Molecular Biology, 2009, 70 (1-2):193-209.

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Tissue Expression Pattern Analysis of TIPs Genes in Phyllostachys edulis

1. International Center for Bamboo and Rattan, State Forestry Administration Key Laboratory on the Science and Technology of Bamboo and Rattan, Beijing 100102, China

Received Date: 2015-08-04

Abstract: [Objective] To reveal the role of tonoplast intrinsic proteins (TIPs) in bamboo under stress conditions and provide new genetic resource for the breeding of new varieties. [Method] The molecular characteristics and expression profiles of TIPs in moso bamboo (Phyllostachys edulis (Carr.) H. de Lehaie) were conducted. The gene expression in different tissues and those under drought, water and NaCl abiotic stresses were analyzed with real-time quantitative PCR. [Result] The result indicated that there were six TIPs homologous genes belonged to three subgroups (TIP1, TIP2 and TIP4) in moso bamboo genome, among which four genes (PeTIP1;1, PeTIP1;2, PeTIP2;2 and PeTIP4;2) consisted of two exons and one intron, and the other two genes (PeTIP2;1 and PeTIP4;1) consisted of three exons and two introns, respectively. Protein structure analysis indicated that all the six PeTIPs had two typical NPA domains and four conserved ar/R selectivity filter. Tissue specific analysis based on transcriptome demonstrated that PeTIP1;1 expressed with high levels in all tissues, PeTIP1;2 had the highest expression level in flower, PeTIP2;1 was mainly expressed in root and rhizome, PeTIP2;2 was specifically expressed in roots, PeTIP4;1 had the highest expression level in leaf, and PeTIP4;2 expressed in shoot and rhizome with high level, but the lowest in the roots. The result of qRT-PCR confirmed that in roots PeTIP4;1 was up-regulated, while PeTIP2;1, PeTIP2;2 and PeTIP4;2 were inhibited significantly (pPeTIP1;1 and PeTIP4;1 increased and those of PeTIP1;2, PeTIP2;2, and PeTIP4;2 decreased significantly (pPeTIPs all raised significantly (pPeTIP1;1, PeTIP1;2 and PeTIP4;1 were up-regulated under drought treatment, all the expression levels of six PeTIPs were significantly increased under water stress (pPeTIP2;1 was suppressed significantly (p[Conclusion] This result indicated that PeTIPs may play roles with varying degrees in bamboo tolerance of drought, water, NaCl and other abiotic stresses.

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[1]	Mueller N D, Gerber J S, Johnston M, et al. Closing yield gaps through nutrient and water management[J], Nature, 2012,490(7419):254-257.
[2]	Kaldenhoff R, Fischer M. Functional aquaporin diversity in plants[J]. Biochimica et Biophysica Acta, 2006, 1758(8):1134-1141.
[3]	Henzler T, Steudle E. Reversible closing of water channels in Chara internodes provides evidence for a composite transport model of the plasma membrane[J]. Journal of Experimental Botany, 1995, 46(2):199-209.
[4]	Chen S, Polle A. Salinity tolerance of Populus[J]. Plant Biology, 2010,12(2):317-333.
[5]	Ruiz-Lozano J M, Porcel R, Azcón C, et al. Regulation by arbuscular mycorrhizae of the integrated physiological response to salinity in plants: new challenges in physiological and molecular studies[J]. Journal of Experimental Botany, 2012, 63(11):4033-4044.
[6]	Maurel C, Tacnet F, Guclu J, et al. Purified vesicles of tobacco cell vacuolar and plasma membranes exhibit dramatically different water permeability and water channel activity[J]. Proceedings of the National Academy of Sciences of the United States of America, 1997, 94(13):7103-7108.
[7]	Regon P, Panda P, Kshetrimayum E, et al. Genome-wide comparative analysis of tonoplast intrinsic protein (TIP) genes in plants[J]. Functional & Integrative Genomics, 2014, 14(4):617-629.
[8]	Sade N, Vinocur B J, Diber A, et al. Improving plant stress tolerance and yield production: is the tonoplast aquaporin SlTIP2;2 a key to isohydric to anisohydric conversion? [J]. New Phytologist, 2009, 181(3):651-661.
[9]	Wang L L, Chen A P, Zhong N Q, et al. The Thellungiella salsuginea tonoplast aquaporin TsTIP1;2 functions in protection against multiple abiotic stresses[J]. Plant Cell Physiology, 2014, 55(1):148-161.
[10]	Peng Y, Lin W, Cai W, et al. Overexpression of a Panax ginseng tonoplast aquaporin alters salt tolerance, drought tolerance and cold acclimation ability in transgenic Arabidopsis plants[J]. Planta, 2007, 226(3):729-740.
[11]	江泽慧.世界竹藤[M].辽宁科学技术出版社,2002,3-10.
[12]	Cui K, He C, Zhang J. Temporal and spatial profiling of internode elongation-associated protein expression in rapidly growing culms of bamboo[J]. Journal of Proteome Research, 2012, 11(4):2492-2507.
[13]	Hepler P K. Calcium: A central regulator of plant growth and development[J]. The Plant Cell, 2005, 17(8):2142-2155.
[14]	Cao K F, Yang S J, Zhang Y J, et al. The maximum height of grasses is determined by roots[J]. Ecology Letters, 2012, 15(7):666-672.
[15]	Nguyen M X, Moon S, Jung K H. Genome-wide expression analysis of rice aquaporin genes and development of a functional gene network mediated by aquaporin expression in roots[J]. Planta, 2013, 238(4):669-681.
[16]	Yue X, Zhao X, Fei Y, et al. Correlation of aquaporins and transmembrane solute transporters revealed by genome-wide analysis in developing maize leaf[J]. Comparative and Functional Genomics, 2012, 2012:546930.
[17]	Zhao H, Peng Z, Fei B, et al. BambooGDB: a bamboo genome database with functional annotation and an analysis platform[J]. Database, 2014, 2014:bau006.
[18]	Gao Z M, Li X P, Li L B, et al. An effective method for total RNA isolation from bamboo[J]. Chinese Forestry Science and Technology, 2006, 5(3):52-54.
[19]	Fan C, Ma J, Guo Q, et al. Selection of reference genes for quantitative real-time PCR in bamboo (Phyllostachys edulis) [J]. PLoS One, 2013, 8(2):e56573.
[20]	Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCt method[J]. Methods, 2001, 25(4):402-408.
[21]	Peng Z, Lu T, Li L, et al. Genome-wide characterization of the biggest grass, bamboo, based on 10,608 putative full-length cDNA sequences[J]. BMC Plant Biology, 2010, 10:116.
[22]	Moore M J, Query C C, Sharp P A. Splicing of precursors to mRNA by the spliceosome[M]. Cold Spring Harbor: Cold Spring Harbor laboratory Press, 1993,303-357.
[23]	Chou K C, Shen H B. Plant-mPLoc: a top-down strategy to augment the power for predicting plant protein subcellular localization[J]. PLoS One, 2010, 5:e11335.
[24]	Wree D, Wu B, Zeuthen T, et al. Requirement for asparagine in the aquaporin NPA sequence signature motifs for cation exclusion[J]. The FEBS Journal, 2011, 278(5):740-748.
[25]	Mitani-Ueno N, Yamaji N, Zhao F J, et al. The aromatic/arginine selectivity filter of NIP aquaporins plays a critical role in substrate selectivity for silicon, boron, and arsenic[J]. Journal of Experimental Botany, 2011, 62(12):4391-4398.
[26]	Reuscher S, Akiyama M, Mori C, et al. Genome-wide identification and expression analysis of aquaporins in tomato[J]. PLoS One, 2013, 8(11):e79052.
[27]	Utsugi S, Shibasaka M, Maekawa M, et al. Control of the water transport activity of barley HvTIP3;1 specifically expressed in seeds[J]. Plant Cell Physiology, 2015, pii:pcv104.
[28]	Forrest K L, Bhave M. The PIP and TIP aquaporins in wheat form a large and diverse family with unique gene structures and functionally important features[J]. Functional & Integrative Genomics, 2008, 8(2):115-133.
[29]	Hove R M, Ziemann M, Bhave M. Identification and expression analysis of the barley (Hordeum vulgare L.) aquaporin gene family[J]. PloS One, 2015, 10(6):e0128025.
[30]	Peng Z H, Lu Y, Li L B, et al. The draft genome of the fast growing non-timber forest species moso bamboo (Phyllostachys heterocycla)[J]. Nature Genetics, 2013, 45(4):456-461.
[31]	Verma R K, Prabh N D, Sankararamakrishnan R. Intra-helical salt-bridge and helix destabilizing residues within the same helical turn: Role of functionally important loop E half-helix in channel regulation of major intrinsic proteins[J]. Biochimica et Biophysica Acta, 2015, 1848(6):1436-1449.
[32]	Zhang D Y, Ali Z, Wang C B, et al. Genome-wide sequence characterization and expression analysis of major intrinsic proteins in soybean (Glycine max L.)[J]. PLoS One, 2013, 8(2):e56312.
[33]	Katsuhara M, Hanba Y T, Shiratake K, et al. Expanding roles of plant aquaporins in plasma membranes and cell organelles[J]. Functional Plant Biology, 2008, 35(1):1-14.
[34]	Schmid M, Davison T S, Henz S R, et al. A gene expression map of Arabidopsis thaliana development[J]. Nature Genetics, 2005, 37(5):501-506.
[35]	Beebo A, Thomas D, Der C, et al. Life with and without AtTIP1;1, an Arabidopsis aquaporin preferentially localized in the apposing tonoplasts of adjacent vacuoles[J]. Plant Molecular Biology, 2009, 70 (1-2):193-209.

Tissue Expression Pattern Analysis of TIPs Genes in Phyllostachys edulis

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通讯作者: 陈斌, bchen63@163.com

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Tissue Expression Pattern Analysis of TIPs Genes in Phyllostachys edulis

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