‫عنوان البحث‬
A DREB gene from the xero-halophyte Atriplex halimus is induced by osmotic but not ionic
stress and shows distinct differences from glycophytic homologues
‫أسماء الباحثين‬
Khedr A.A.,
Serag M.M.,
Nemat Alla M.M.,
Abu-Alnaga A.Z.
Nada R.M.
Paul Quick W.
Abogadallah G.M.
‫بيانات الباحثين‬
Botany Department, Faculty of Science, Mansoura University, Damietta, Egypt
Abstract
Studying the regulation of stress inducible genes can lead to understanding of the mechanisms by which
halophytes maintain growth and thrive under abiotic stress. Verifying whether ionic or osmotic components of
salt stress control the regulation of Dehydration Responsive Element- binding transcription factor (DREB) was
investigated in the present study. DREB belongs to AP2/EREB group that is induced under abiotic stress and
regulates many stress inducible genes that contain DRE binding sites in their promoters. DREB in Atriplex
halimus was regulated by the osmotic component but not by the ionic one of salt stress. It seemed that DREB
was not involved in the regulation of sodium manipulating genes like NHX1, SOS1 or H?-PPase. Moreover,
DREB could be involved directly of indirectly in CMO regulation because of timing of induction. Also, DREB
was the most upregulated gene under salt (fivefold) and drought (twofold) conditions, which reinforced the
importance of this gene in A. halimus tolerance to stress. Moreover, its constitutive expression under normal
conditions also indicated its involvement in other growth and developmental programs. The tolerance of A.
halimus and of halophytes, in general, could be attributed to the constitutive expression of its genes and
differences in protein structures between halophytes and glycophytes.
Keywords:
Atriplex halimus, DREB, Protein content, Gene expression, Salinity, Osmotic and ionic stress
Published In:
Plant Cell Tiss Organ Cult 106:191–206 (2011)
References
Allen MD, Yamasaki M, Ohme-Takagi M, Tatenol M, Suzuki M (1998) A novel mode of DNA recognition by
a b-sheet revealed by the solution structure of the GCC-box binding-domain in complex with DNA. Eur
Mol Biol Organ J 17:5484–5496
Apse MP, Blumwald E (2002) Engineering salt tolerance. Cur Opin Biotech 13:146–150
Apse MP, Aharon GS, Snedden WA, Blumwald E (1999) Salt tolerance conferred by overexpression of a
vacuolar Na?/H? antiport in arabidopsis. Sci 285:1256–1258
Bajji M, Kinet JM, Lutts S (1998) Salt stress effects on roots and leaves of Atriplex halimus L. and their
corresponding callus cultures. Plant Sci 137:131–142
Ben Hassine A, Ghanem ME, Bouzid S, Lutts S (2008) An inland and a coastal population of the
Mediterranean xero-halophyte species Atriplex halimus L. differ in their ability to accumulate proline and
glycinebetaine in response to salinity and water stress. J Exp Bot 59:1315–1326
Blumwald E, Aharon GS, Apse MP (2000) Sodium transport in plant cells. Bioch Biophy Acta 1456:140–151
Cao ZF, Li J, Chen F, Li YQ, Zhou HM, Liu Q (2001) Effect of two conserved amino-acid residues on
DREB1A function. Biochem 66:623–627
Chen M, Wang QY, Cheng XG, Xu ZS, Li LC, Ye XG, Xia lQ, Ma YZ (2007) GmDREB2, a soybean DREbinding transcription factor, conferred drought and high-salt tolerance in transgenic plants. Bioch Biophy
Res Commu 353:299–305
Chen J, Xia X, Yin W (2009) Expression profiling and functional characterization of a DREB2-type gene from
Populus euphratica. Bioch Biophy Res Commu 378:483–487
Cong L, Zheng HC, Zhag YX, Chai TY (2008) Arabidopsis DREB1A confers high salinity tolerance and
regulates the expression of GA dioxygenases in Tobacco. Plant Sci 174:156–164
Diedhiou CJ, Popova OV, Golldack D (2009) Transcript profiling of the salt-tolerant Festuca rubra ssp.
litoralis reveals a regulatory network controlling salt acclimatization. J Plant Physiol 166: 697–711
Fowler S, Thomashow MF (2002) Arabidopsis transcriptome profiling indicates that multiple regulatory
pathways are activated during cold acclimation in addition to the CBF cold response pathway. Plant Cell
14:1675–1690
Gao SQ, Chen M, Xia LQ, Xiu HJ, Xu ZH, Li LC, Zhao CP, Cheng XC, Ma YZ (2009) A cotton (Gossypium
hirsutum) DREbinding transcription factor gene, GhDREB, confers enhanced tolerance to drought, high
salt, and freezing stresses in transgenic wheat. Plant Cell Rep 28:301–311
Gaxiola RA, Li J, Undurraga S, Dang LM, Allen GJ, Alper SL, Fink GR (2001) Drought- and salt-tolerant
plants result from overexpression of the AVP1 H1-pump. Pro Nat Acad Sci USA 98:11444–11449
Gilmour SJ, Zarka DG, Stockinger EJ, Salazar MP, Houghton JM, ThomashowMF (1998) Low temperature
regulation of the ArabidopsisCBF family ofAP2 transcriptional activators as an early step in cold-induced
COR gene expression. Plant J 16:433–443
Gilmour SJ, Sebolt AM, Salazar MP, Everard JD, Thomashow MF (2000) Overexpression of the Arabidopsis
CBF3 transcriptional activator mimics multiple biochemical changes associated with cold acclimation.
Plant Physiol 124:1854–1865
Gilmour SJ, Fowler SG, Thomashow MF (2004) Arabidopsis transcriptional activators CBF1, CBF2, and
CBF3 have matching functional activities. Plant Mol Biol 54:767–781
Glenn EP, Brown JJ, Blumwald E (1999) Salt tolerance and crop potential oh halophytes. Cri Rev Plant Sci
18:227–255
Hansen EH, Munns DN (1988) Effect of CaSO4 and NaCl on mineral content of Leucaena leucocephala. Plant
Soil 7:101–105
Hasegawa PM, Bressan RA, Zhu JK, Bohnert HJ (2000) Plant cellular and molecular responses to high
salinity. Ann Rev Plant Physiol Plant Mol Biol 51:463–499
Hatch MD, Osmond CB (1976) Compartmentation and transport in C4 photosynthesis. In: Stocking CR, Heber
U (eds) Transport in plants. III. Intracellular interactions and transport processes. Encyclopedia of plant
physiology, New Series 3 pp 144–184
Hayashi H, Murata N (1998) Genetically engineered enhancement of salt tolerance in higher plants. In: Satoh
K, Murata N (eds) Stress responses of photosynthetic organisms. Elsevier, Amsterdam, pp 133–148
Hewitt EJ (1966) Sand and water culture methods used in the study of plant nutrition. Technical
communication 22. Commonwealth Bureau of horticulture plantation crops, GB Hunte C, Screpanti E,
Venturi M, Rimon A, Padan E, Michel H (2005) Structure of a Na?/H? antiporter and insights into
mechanism of action and regulation by pH. Nat 435:1197–1202
Hurkman WJ, Tanaka CK (1986) Solubilization of plant membrane proteins for analysis by two-dimensional
gel electrophoresis. Plant Physiol 81:802–806
Ito Y, Katsura K, Maruyama K, Taji T, Kobayashi M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2006)
Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene
expression in transgenic rice. Plant Cell Physiol 47:141–153
Jin T, Chang Q, Li W, Yin D, Li Z, Wang D, Liu B, Liu L (2010) Stress-inducible expression of GmDREB1
conferred salt tolerance in transgenic alfalfa. Plant Cell Tiss Organ Cult 100: 219–227
Kant S, Kant P, Raveh E, Barak S (2006) Evidence that differential gene expression between the halophyte,
Thellungiella halophila, and Arabidopsis thaliana is responsible for higher levels of the compatible
osmolyte proline and tight control of Na? uptake in T halophila. Plant Cell Environ 29:1220–1234
Knight H, Knight MR (2001) Abiotic stress signalling pathways: specificity and cross-talk. Trends Plant Sci
6:262–267
Laemmlli EK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nat
227:680–685
Leubner-Metzger G, Petruzzelli L, Waldvogel R (1998) Ethylene responsive element binding protein (EREBP)
expression and the transcriptional regulation of class I beta-1, 3-glucanase during tobacco seed
germination. Plant Mol Biol 38:785–795
Li J, Yang H, Peer WA, Richter G, Blakeslee J, Bandyopadhyay A, Titapiwantakun B, Undurraga S,
Khodakovskaya M, Richards EL, Krizek BA, Murphy AS, Gilroy S, Gaxiola RA (2005) Arabidopsis H?PPase AVP1 regulates auxin mediated organ development. Sci 310:121–125
Li JY, He X, Xu L, Zhou J, Wu P, Shou H, Zhang F (2008) Molecular and functional comparisons of the
vacuolar Na?/H? exchangers originated from glycophytic and halophytic species. J Zhejiang Univ 9:132–
140
Li HY, Wang YC, Jiang J, Liu GF, Gao CQ, Yang CP (2009) Identification of genes responsive to salt stress
on Tamarix hispida roots. Gene 433:65–71
Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (1998) Two transcription
factors, DREB1 and DREB2, with an EREBP/AP2 DNA binding domain separate two cellular signal
transduction pathways in drought- and lowtemperature- responsive gene expression, respectively, in
Arabidopsis. Plant Cell 10:1391–1406
Liu Y, Liu J, Tang MJ, Diao Y, Shen SH (2005) A DREB gene in asparagus. Photosynthesis Research Center,
Institute of Botany, Chinese Academy of Science, China
Lokhande VH, Nikam TD, Patade VY, Ahire ML, Suprasanna P (2010) Effects of optimal and supra-optimal
salinity stress on antioxidative defence, osmolytes and in vitro growth responses in Sesuvium
portulacastrum L. Plant Cell Tiss Organ Cult. doi: 10.1007/s11240-010-9802-9
Ma XL, Zhang Q, Shi HZ, Zhu JK, Zhao YX, Ma CL, Zhang H (2004) Molecular cloning and different
expression of a vacuolar Na?/H? antiporter gene in Suaeda salsa under salt stress. Biol Plant 48:219–225
Maeshima M (2000) Vacuolar H?-pyrophosphatase. Bioch Biophy Acta 1465:37–51
Maeshima M (2001) Tonoplast transporters: organization and function. Ann Rev Plant Physiol Plant Mol Biol
52:469–497
Martınez JP, Ledent JF, Bajji M, Kinet JM, Lutts S (2003) Effect of water stress on growth, Na and K
accumulation and water use efficiency in relation to osmotic adjustment in two populations of Atriplex
halimus L. Plant Grow Regu 41:63–73
Martınez JP, Kinet JM, Bajji M, Lutts S (2005) NaCl alleviates polyethylene glycol-induced water stress in the
halophyte species Atriplex halimus L. J Exp Bot 56:2421–2431
Maruyama K, Sakuma Y, Kasuga M, Ito Y, Seki M, Goda H, Shimada Y, Yoshida S, Shinozaki K, YamaguchiShinozaki K (2004) Identification of cold-inducible downstream genes of the Arabidopsis
CBF3/DREB1A transcriptional factor using two microarray systems. Plant J 38:982–993
Matsuoka M, Furbank RT, Fukayama H, Miyao M (2001) Molecular engineering of C4 photosynthesis. Ann
Rev Plant Physiol Plant Mol Biol 52:297–314
Munns R (1993) Physiological processes limiting plant growth in saline soils: some dogmas and hypotheses.
Plant Cell Environ 16:15–22
Munns R, Tester M (2008) Mechanisms of salinity tolerance. Ann Rev Plant Biol 59:651–681
Murata S, Kobayashi M, Matoh T, Sekiya J (1992) Sodium stimulates regeneration of phosphoenolpyruvate in
mesophyll chloroplasts of chloroplasts of Amaranthus tricolor. Plant Cell Physiol 33:1247–1250
Nakashima K, Yamaguchi-Shinozak K (2006) Regulons involved in osmotic stress-responsive and cold stressresponsive gene expression in plants. Physiol Plant 126:62–71
Niu X, Bressan RA, Hasegawa PM, Pardo JM (1995) Ion homeostasis in NaCl stress environments. Plant
Physiol 109:735–742
Oh SJ, Song SI, Kim YS, Jang HJ, Kim SY, Kim M, Kim YK, Nahm BH, Kim JK (2005) Arabidopsis
CBF3/DREB1A and ABF3 in transgenic rice increased tolerance to abiotic stress without stunting
growth. Plant Physiol 138:341–351
Okamuro JK, Caster B, Villarroel R, Montagu MV, Jofuku KD (1997) The AP2 domain of APETALA2
defines a large new family of DNA binding-proteins in Arabidopsis. Pro Nat Acad Sci USA 94:7076–
7081
Pellegrineschi A, Reynolds M, Pacheco M, Brito RM, Almeraya R, Yamaguchi-Shinozaki K, Hoisington D
(2004) Stress-induced expression in wheat of the Arabidopsis thaliana DREB1A gene delays water stress
symptoms under greenhouse conditions. Gen 47:493–500
Qin F, Li J, Zhang GY, Zhao J, Chen SY, Liu Q (2003) Isolation and structural analysis of DRE-binding
transcription factor from maize (Zea mays L.). Acta Botin Sini 45:331–339
Rea PA, Poole RJ (1993) Vacuolar H?-translocating pyrophosphatase. Ann Rev Plant Physiol Plant Mol Biol
44:157–180
Rhodes D, Hanson AD (1993) Quaternary ammonium and tertiary sulfonium compounds in higher plants.
Annu Rev Plant Physiol Plant Mol Biol 44:357–384
Seki M, Narusaka M, Abe H, Kasuga M, Yamaguchi-Shinozaki K, Carninci P, Hayashizaki Y, Shinozaki K
(2001) Monitoring the expression pattern of 1300 Arabidopsis genes under drought and cold stresses by
using a full-length cDNA microarray. Plant Cell 13:61–72
Shen YG, Zhang WK, He SJ, Zhang ZS, Liu Q, Chen SY (2003a) An EREBP/AP2-type protein in Triticum
aestivum was a DREbinding transcription factor induced by cold, dehydration, and ABA stress. Theor
Appl Gene 106:923–930
Shen YG, Zhang WK, Yan DQ, Du BX, Zhang ZS, Liu Q, Chen SY (2003b) Characterization of a DREbinding transcription factor from a halophyte Atriplex hortensis. Theor Appl Gene 107:155–161
Shi H, Zhu JK (2002) Regulation of expression of the vacuolar Na?/ H? antiporter gene AtNHX1 by salt stress
and abscisic acid. Plant Mol Biol 50:543–550
Subbarao GV, Wheeler RM, Levine LH, Stutte GW (2001) Glycine betaine accumulation, ionic and water
relations of red beet at contrasting levels of sodium supply. J Plant Physiol 158:767–776
Subramanyam K, Sailaja KV, Subramanyam K, Rao DM, Lakshmidevi K (2010) Ectopic expression of an
osmotin gene leads to enhanced salt tolerance in transgenic chilli pepper (Capsicum annum L.). Plant Cell
Tiss Organ Cult. doi: 10.1007/s11240- 010-9850-1
Sze H, Wald JM, Lai S, Perera I (1992) Vacuolar-type H?- translocating ATPases in plant endomembranes:
subunit organization and multigene families. J Exp Biol 172:123–135
Taji T, Seki M, Satou M, Sakurai T, Kobayashi M, Ishiyama K, Narusaka Y, Narusaka M, Zhu JK, Shinozaki
K (2004) Comparative genomics in salt tolerance between arabidopsis and arabidopsis-related halophyte
salt cress using arabidopsis microarray. Plant Physiol 135:1697–1709
Takahashi R, Liu S, Takano T (2009) Isolation and characterization of plasma membrane Na?/H? antiporter
genes from salt-sensitive and salt-tolerant reed plants. J Plant Physiol 166:301–309
Wang YM, He CF (2007) Isolation and characterization of a coldinduced DREB gene from Aloe Vera L. Plant
Mol Biol Rep 25:121–132
Wang BS, Luttge U, Ratajczak R (2001) Effects of salt treatment and osmotic stress on V-ATPase and VPPase in leaves of the halophyte Suaeda salsa. J Exp Bot 52:2355–2365
Wang ZL, Li PH, Fredricksen M, Gong ZZ, Kimd CS, Zhang CQ, Bohnert HJ, Zhu JK, Bressan RA, Hasegawa
PM, Zhao YX, Zhang H (2004) Expressed sequence tags from Thellungiella halophila, a new model to
study plant salt-tolerance. Plant Sci 166:609–616
Wang Q, Guan Y, Wu Y, Chen H, Chen F, Chu C (2008) Overexpression of a rice OsDREB1F gene increases
salt, drought, and low temperature tolerance in both Arabidopsis and rice. Plant Mol Biol 67:589–602
Wang QJ, Xu KY, Tong ZG, Wang SH, Gao ZH, Zhang JY, Zong CW, Qiao YS, Zhang Z (2010)
Characterization of a new dehydration responsive element binding factor in central arctic cowberry. Plant
Cell Tiss Organ Cult 101:211–219
Wei Q, Guo YJ, Cao HM, Kuai BK (2010) Cloning and characterization of an AtNHX2-like Na?/H? antiporter
gene from Ammopiptanthus mongolicus (Leguminosae) and its ectopic expression enhanced drought and
salt tolerance in Arabidopsis thaliana. Plant Cell Tiss Organ Cult. doi: 10.1007/s11240- 010-9869-3
Wu Y, Ding N, Zhao X, Zhao M, Chang Z, Liu J, Zhang L (2007) Molecular characterization of PeSOS1: the
putative Na?/H? antiporter of Populus euphratica. Plant Mol Biol 65:1–11
Xiong LM, Zhu JK (2002) Salt tolerance. In: Somerville CR, Meyerowitz EM (eds) The Arabidopsis book.
American Society of Plant Biologists, Rockville, pp 1–22
Yamaguchi-Shinozaki K, Shinozaki K (1994) A novel cis-acting element in an Arabidopsis gene is involved in
responsiveness to drought, low temperature, or high-salt stress. Plant Cell 6:251–264
Yamaguchi-Shinozaki K, Shinozaki K (2006) Transcriptional regulatory networks in cellular response and the
tolerance to dehydration and cold stresses. Ann Rev Plant Biol 57:781–803
Yeo ER (1998) Molecular biology of salt tolerance in the context of whole plant physiology. J Exp Bot
49:915–929
Yoshiba Y, Kiyosue T, Nakashima K, Yamaguchi-Shinozaki K, Shinozaki K (1997) Regulation of levels of
proline as an osmolyte in plants under water stress. Plant Cell Physiol 38: 1095–1102
Zhang L, Ma XL, Zhang Q, Ma CL, Wang PP, Sun YF, Zhao YX, Zhang H (2001) Expressed sequence tags
from a NaCl-treated Suaeda salsa cDNA library. Gene 267:193–200
Zhou JM, Tang XY, Martin GB (1997) The Pto kinase conferring resistance to tomato bacterial speck disease
interacts with proteins that bind a cis-element of pathogenesis-related genes. Eur Mol Biol Organ J
16:3207
Zhu JK (2001) Plant salt tolerance. Trends Plant Sci 6:66–71
Zhu JK (2002) Salt and drought stress signal transduction in plants. Ann Rev Plant Biol 53:247–273