Cloning and expression analysis of a homologous expansin gene EXP2 in Picea wilsonii

Abstract

Expansins are cell-wall-loosening proteins that have multiple roles during plant development and stress-related processes. In this study, a novel expansin gene PwEXP2 was cloned by rapid amplification of cDNA ends based on the cDNA library of Picea wilsonii and EST fragment of PwEXP2. It was found that PwEXP2 coded 253 amino acids, and putative signal peptides exist at the N-terminal, followed by 8 cysteines, a HFD (His-Phe-Asp) conserved domain, and 4 tryptophan residues at the C-terminal. PwEXP2 was located in cytoplasm and nucleus when transformed in an onion epidermal cell. Quantitative real-time PCR assays showed that PwEXP2 was expressed in various tissues with a relatively high level in needles and low level in mature pollen. The expression level of PwEXP2 dramatically increased after seed germination. Gene expression profiles in abiotic stresses showed that PwEXP2 was induced by high temperature and osmotic stress but not involved in ABA-dependent signaling pathway. These results display the important roles of the PwEXP2 in plant development and multiple adversity stresses.

Keywords

Expansin / Picea wilsonii / Stress response / Gene expression

References

[1] Beaulieu J, Giguère I, Deslauriers M, Boyle B, MacKay J. Differential gene expression patterns in white spruce newly formed tissue on board the International Space Station. Adv Space Res, 2013, 52: 760-772.

[2] Chen F, Dahal P, Bradford KJ. Two tomato expansin genes show divergent expression and localization in embryos during seed development and germination. Plant Physiol, 2001, 127: 928-936.

[3] Cho HT, Cosgrove DJ. Regulation of root hair initiation and expansin gene expression in Arabidopsis. Plant Cell Online, 2002, 14: 3237-3253.

[4] Dotto MC, Martínez GA, Civello PM. Expression of expansin genes in strawberry varieties with contrasting fruit firmness. Plant Physiol Biochem, 2006, 44: 301-307.

[5] Gao X, Liu K, Lu YT. Specific roles of AtEXPA1 in plant growth and stress adaptation. Russ J Plant Physiol, 2010, 57: 241-246.

[6] Geilfus CM, Zörb C, Neuhaus C, Hansen T, Lüthen H, Mühling KH. Differential transcript expression of wall-loosening candidates in leaves of maize cultivars differing in salt resistance. J Plant Growth Regul, 2011, 30: 387-395.

[7] Georgelis N, Yennawar NH, Cosgrove DJ. Structural basis for entropy-driven cellulose binding by a type-A cellulose-binding module (CBM) and bacterial expansin. Proc Natl Acad Sci, 2012, 109: 14830-14835.

[8] Han YY, Li AX, Li F, Zhao MR, Wang W. Characterization of a wheat (Triticum aestivum L.) expansin gene, TaEXPB23, involved in the abiotic stress response and phytohormone regulation. Plant Physiol Biochem, 2012, 54: 49-58.

[9] Hayama H, Ito A, Moriguchi T, Kashimura Y. Identification of a new expansin gene closely associated with peach fruit softening. Postharvest Biol Technol, 2003, 29: 1-10.

[10] Hernández-Nistal J, Labrador E, Martín I, Jiménez T, Dopico B. Transcriptional profiling of cell wall protein genes in chickpea embryonic axes during germination and growth. Plant Physiol Biochem, 2006, 44: 684-692.

[11] Hiwasa K, Rose JK, Nakano R, Inaba A, Kubo Y. Differential expression of seven α-expansin genes during growth and ripening of pear fruit. Physiol Plant, 2003, 117: 564-572.

[12] Huang J, Takano T, Akita S. Expression of α-expansin genes in young seedlings of rice (Oryza sativa L.). Planta, 2000, 211: 467-473.

[13] Kende H, Bradford KJ, Brummell DA, Cho HT, Cosgrove DJ, Fleming AJ, Gehring C, Lee Y, McQueen-Mason S, Rose JKC, Voesenek LACJ. Nomenclature for members of the expansin superfamily of genes and proteins. Plant Mol Biol, 2004, 55: 311-314.

[14] Kwon YR, Lee HJ, Kim KH, Hong SW, Lee SJ, Lee H. Ectopic expression of Expansin3 or Expansinβ1 causes enhanced hormone and salt stress sensitivity in Arabidopsis. Biotechnol Lett, 2008, 30: 1281-1288.

[15] Li F, Wang W. Characterization of a wheat (Triticum aestivum L.) expansin gene, TaEXPB23, involved in the abiotic stress response and phytohormone regulation. Plant Physiol Biochem, 2012, 54: 49-58.

[16] Li Y, Jones L, McQueen-Mason S. Expansins and cell growth. Curr Opin Plant Biol, 2003, 6: 603-610.

[17] Li HY, Jiang J, Wang S, Liu FF. Expression analysis of ThGLP, a new germin-like protein gene, in Tamarix hispida. J For Res, 2010, 21: 323-330.

[18] Li F, Han Y, Feng Y, Xing S, Zhao M, Chen Y, Wang W. Expression of wheat expansin driven by the RD29 promoter in tobacco confers water-stress tolerance without impacting growth and development. J Biotechnol, 2013, 163: 281-291.

[19] Lü P, Kang M, Jiang X, Dai F, Gao J, Zhang C. RhEXPA4, a rose expansin gene, modulates leaf growth and confers drought and salt tolerance to Arabidopsis. Planta, 2013, 237: 1547-1559.

[20] Mbéguié-A-Mbéguié D, Gouble B, Gomez RM, Audergon JM, Albagnac G, Fils-Lycaon B. Two expansin cDNAs from Prunus armeniaca expressed during fruit ripening are differently regulated by ethylene. Plant Physiol et Biochem, 2002, 40: 445-452.

[21] McQueen-Mason S, Durachko DM, Cosgrove DJ. Two endogenous proteins that induce cell wall extension in plants. Plant Cell Online, 1992, 4: 1425-1433.

[22] Nakamura Y, Wakabayashi K, Hoson T. Temperature modulates the cell wall mechanical properties of rice coleoptiles by altering the molecular mass of hemicellulosic polysaccharides. Physiol Plant, 2003, 118: 597-604.

[23] Sasayama D, Azuma T, Itoh K. Changes in expansin activity and cell wall susceptibility to expansin action during cessation of internodal elongation in floating rice. Plant Growth Regul, 2009, 57: 79-88.

[24] Sasidharan R, Voesenek LACJ, Pierik R. Cell wall modifying proteins mediate plant acclimatization to biotic and abiotic stresses. Crit Rev Plant Sci, 2011, 30: 548-562.

[25] Shao HB, Guo QJ, Chu LY, Zhao XN, Su ZL, Hu YC, Cheng JF. Understanding molecular mechanism of higher plant plasticity under abiotic stress. Colloids Surfaces B-Biointerfaces, 2007, 54: 37-45.

[26] Takahashi R, Fujitani C, Yamaki S, Yamada K. Analysis of the cell wall loosening proteins during rose flower opening. Int Conf Qual Manag Supply Chains Ornam, 2007, 755: 483-488.

[27] Vreeburg RA, Benschop JJ, Peeters AJ, Colmer TD, Ammerlaan AH, Staal M, Elzenga TM, Staals RH, Darley CP, McQueen-Mason SJ, Voesenk LA. Ethylene regulates fast apoplastic acidification and expansin A transcription during submergence-induced petiole elongation in Rumex palustris. Plant J, 2005, 43: 597-610.

[28] Wei PC, Chen S, Zhang XQ, Zhao P, Xiong YM, Wang WL, Chen J, Wang XC. An α-expansin, VfEXPA1, is involved in regulation of stomatal movement in Vicia faba L.. Chin Sci Bull, 2011, 56: 3531-3537.

[29] Xu X, Huang BR, Xu JC. Analysis of AsEXP1 gene related to drought tolerance in creeping bentgrass. J Beijing For Univ, 2010, 32: 126-131.

[30] Yamada K, Takahashi R, Fujitani C, Mishima K, Yoshida M, Joyce DC, Yamaki S. Cell wall extensibility and effect of cell-wall-loosening proteins during rose flower opening. J Jpn Soc Hortic Sci, 2009, 78: 242-251.

[31] Yu YL, Li YZ, Huang GX, Meng ZD, Zhang D, Wei J, Yan K, Zheng CC, Zhang LY. PwHAP5, a CCAAT-binding transcription factor, interacts with PwFKBP12 and plays a role in pollen tube growth orientation in Picea wilsonii. J Exp Bot, 2011, 62: 4805-4817.

[32] Zenoni S, Fasoli M, Tornielli GB, Santo SD, Sanson A, de Groot P, Sordo S, Citterio S, Monti F, Pezzotti M. Overexpression of PhEXPA1 increases cell size, modifies cell wall polymer composition and affects the timing of axillary meristem development in Petunia hybrida. New Phytol, 2011, 191: 662-677.

[33] Zhang D, Liu YJ, Li CJ, Cao YB, Zhang LY. Construction of normalized cDNA library and analysis of corresponding EST sequences in Picea wilsonii. Biotechnol Bull, 2012, 6: 71-76.

[34] Zhao MR, Han YY, Feng YN, Li F, Wang W. Expansins are involved in cell growth mediated by abscisic acid and indole-3-acetic acid under drought stress in wheat. Plant Cell Rep, 2012, 31: 671-685.

[35] Zhou P, Zhu Q, Xu J, Huang B. Cloning and characterization of a gene, encoding expansin proteins inducible by heat stress and hormones in creeping bentgrass. Crop Sci, 2011, 43: 333-341.