Cloning and Expression Analysis of Promoter of GRAS Transcription Factor from Tamarix hispida

doi:10.11924/j.issn.1000-6850.casb15080006

Abstract

Abstract: In order to explore the expression characteristics and functions of GRAS transcription factor gene in Tamarix hispida, the promoter sequence of GRAS transcription factor was cloned and its expression patterns were analyzed. The genomic DNA of T. hispida was extracted by using CTAB method. The promoter sequence of GRAS gene was cloned by using the Genome Walking Kit according to the instructions. The 35S promoter sequence of the pCAMBIA1301 expression vector was replaced by the GRAS promoter sequence to construct the fusion expression vectors and drove the expression of GUS gene. Then, the fusion expression vectors were transiently expressed in Arabidopsis thaliana and the transformed seedlings were stained by GUS. The 936 bp gene promoter of GRAS transcription factor was cloned from Tamarix hispida. The analysis results by using the PLACE and PlantCARE software suggested that the promoter sequences contained not only the core elements such as CAAT- box and TATA- box, but also some cis- element related to the stress response. The DNA recombinant plasmid PGRAS::GUS was constructed successfully by using the GRAS promoter sequence to replace the 35S promoter sequence of pCAMBIA1301, and was transiently transferred into Arabidopsis. The results of GUS staining showed that the leaves of transformed Arabidopsis were stained while the roots were stained slightly. These results indicated that the GRAS gene promoter sequence had promoter expression activity, and it might play a role in the stress response of T. hispida. The results provided the foundation for a comprehensive analysis of the stress response function and mechanisms of GRAS gene in T. hispida.

References

[1] Pysh LD,Wysocka-Diller JW,Camilleri C,Bouchez D,Benfey PN (1999). The GRAS gene family in Arabidopsis: sequence characterization and basic expression analysis of the SCARECROW-LIKE genes. Plant J.18(1):111-119
[2] Hirsch S, Kim J, Mu?oz A, Heckmann AB, Downie JA, Oldroyd GE(2009). GRAS proteins form a DNA binding complex to induce gene expression during nodulation signaling in Medicago truncatula. Plant Cell, 21(2): 545~557
[3] Aleman L, Kitamura J, Abdel-mageed H, Lee J, Sun Y, Nakajima M,Ueguchi-Tanaka M, Matsuoka M, Allen RD (2008). Functional analysis of cotton orthologs of GA signal transduction factors GID1 and SLR1. Plant Mol Biol, 68(1-2): 1~16
[4] Zhang ZL, Ogawa M, Fleet CM, Zentella R, Hu J, Heo JO, Lim J, Kamiya Y, Yamaguchi S, Sun TP (2011). Scarecrow-like 3 promotes gibberellin signaling by antagonizing master growth repressor DELLA in Arabidopsis. Proc Natl Acad Sci USA, 108(5): 2160~2165
[5] Bolle C, Koncz C, Chua NH (2000). PAT1, a new member of the GRAS family, is involved in phytochrome A signal transduction. Genes Dev, 14(10): 1269~1278
[6] Gleason C, Chaudhuri S, Yang T, Munoz A, Poovaiah BW, Oldroyd GE (2006). Nodulation independent of rhizobia induced by a calcium-activated kinase lacking autoinhibition. Nature, 441(7097):1149~1152
[7] Torres-Galea P, Hirtreiter B, Bolle C (2013). Two GRAS proteins,SCARECROW-LIKE21 and PHYTOCHROME A SIGNAL TRANSDUCTION1, function cooperatively in phytochrome A signal transduction. Plant Physiol, 161(1): 291~304
[8] Tian C, Wan P, Sun S, Li J, Chen M (2004) . Genome-wide analysis of the GRAS gene family in rice and Arabidopsis . Plant Mol Biol, 54(4):519–532.
[9] Li X, Qian Q, Fu Z, Wang Y, Xiong G, Zeng D, Wang X, Liu X, Teng S, Hiroshi F, Yuan M, Luo D, Han B, Li J (2003).Control of tillering in rice. Nature, 422(6932): 618-621.
[10] Lee MH, Kim B, Song SK, Heo JO, Yu NI, Lee SA, Kim M, Kim DG, Sohn SO, Lim CE, Chang KS, Lee MM, Lim J (2008). Large-scale analysis of the GRAS gene family in Arabidopsis thaliana. Plant Mol Biol 67(6), 659-670.
[11] Potenza C, Aleman L, Sengupta-Gopalan C (2004). Targeting transgene expression in research, agricultural, and environmental applications: promoters used in plant transformation. In Vitro Cell Dev Biol Plant, 40(1):1-22
[12] Gowik U, Burscheidt J, Akyildiz M, Schlue U, Koczor M, Streubel M, Westhoff P (2004). cis-Regulatory elements for mesophyll-specific gene expression in the C4 plant flaveria trinervia, the promoter of the C4 phosphoenolpyruvate carboxylase gene.Plant cell, 16(5):1077-1090.
[13]Mitsutakataniguchi KatsuraizawA ,Maurice SB (2000).The Promoter for the maize C4 Pyruvate,orthophosphate dikinase gene directs cell and tissue-specific transcription in transgenic maize plants. Plant Cell Physiol, 41(1):42-48
[14]Montgomery J,Goldman S,Deikman J,Margossina LandFiseher RL(1993). Identification of an ethylene-responsive region in the Promoter of a fruit ripening gene. Proc Natl Acad Sci USA. 90(13):5939-5943
[15] Schernthaner JP, Matzke MA, Matzke AJ (1998). Endosperm-specific activity of a zein gene promoter in transgenic tobacco plants. EMBO J, 7(5):1249–1255.
[16] Colot V, Robert LS, Kavanagh TA, Bevan MW, Thompson RD (1987). Localization of sequences in wheat endosperm protein genes which confer tissue-specific expression in tobacco. EMBO J, 6(12):3559-3564.
[17] Leisy DJ, Hnilo J, Zhao Y, Okita TW (1990). Expression of a rice glutelin promoter in transgenic tobacco [J].Plant Mol Biol , 14(1):41–50.
[18] Wobus U, Borisjuk L, Panitz R, Manteuffel R, B?umlein H, Wohlfahrt T, Heim U, Weber H, Miséra S, Weschke W(1995).Control of seed storage protein gene expression: new aspects on an old problem[J]. J Plant Physiol, 145(5-6):592–599.
[19] Saidi Y, Finka A, Chakhporanian M, Zr?d JP, Schaefer DG, Goloubinoff P (2005). Controlled expression of recombinant proteins in Physcomitrella patens by a conditional heat-shock promoter: a tool for plant research and biotechnology. Plant Mol Biol, 59(5):697–711.
[20] Zheng L, Liu G, Meng X, Liu Y, Ji X, Li Y, Nie X, Wang Y(2013)．A WRKY gene from Tamarix hispida, ThWRKY4, mediates abiotic stress responses by modulating reactive oxygen species and expression of stress-responsive genes.Plant Mol Biol. 82(4-5): 303-320
[21] Ma HS, Liang D, Shuai P, Xia XL, Yin WL (2010). The salt- and drought-inducible poplar GRAS protein SCL7 confers salt and drought tolerance in Arabidopsis thaliana. J Exp Bot, 61(14):4011~4019
[22] Kaur C, Kumar G, Kaur S, Ansari MW, Pareek A, Sopory SK, Singla-Pareek SL (2015). Molecular cloning and characterization of salt overly sensitive gene promoter from Brassica juncea (BjSOS2). Mol Biol Rep. 42(6): 1139-1148.
[23] Shen Z, Yao J, Sun J, Chang L, Wang S, Ding M, Qian Z, Zhang H, Zhao N, Sa G, Hou P, Lang T, Wang F, Zhao R, Shen X, Chen S (2015).. Populus euphratica HSF binds the promoter of WRKY1 to enhance salt tolerance. Plant Science, 235 89–100