532 Plant Mol Biol Rep (2011)29:525-532 (Chinnusamy et al.2010).Does non-heading Chinese Jiang FL,Hou XL,Shi GJ,Cui XM (2007b)Cloning and cabbage have pre-mRNA processing in cold acclimation? characterization of full length cDNA of BrCOR/4 gene from Further studies also need to be carried out. Brassica campestris ssp.chinensis.Jiangsu J of Agr Sci 23:34- 38 (in Chinese) Jiang FL,Hou XL,Shi GJ,Cui XM(2007c)Cloning and characterization Acknowledgements This research was partially supported by the of full length cDNA of BrLOS2 gene from Brassica campestris ssp Natural Science Foundation of Jiangsu Province (BK2009311)and chinensis.J Nanjing Agri Uni 30:27-32 (in Chinese) National Science and Technology Support Program(2009BADB8B03-1). Kasuga M,Liu Q,Miura S,Yamaguchi-Shinozaki K,Shinozaki K (1999)Improving plant drought,salt,and freezing tolerance by gene transfer of a single stress-inducible transcription factor.Nat References Biotechnol 17:287-291 Lee B,Henderson DA,Zhu JK (2005)The arabidopsis cold responsive transcriptome and its regulation by ICEl.Plant Cell Altus NN,Uemura M,Steponkus PL,Gilmour SJ,Lin C,Thomashow 17:3155-3175 MF(1996)Constiutive expression of the cold-regulated Arabidopsis Liu Q,Kasuga M,Sakuma Y,Abe H,Miura S,Yamaguchi-Shinozaki K, thaliana COR15a gene affects both chloroplast and protoplast Shinozaki K(1998)Two transcription factors,DREBI and DREB2 freezing tolerance.Plant Biol 93:13404-13409 with an EREBP/AP2 DNA binding domain separate two cellular Buttner M.Singh KB (1997)Arabidopsis thaliana ethylene respon- signal transduction pathways in drought-and low-temperature- sive element binding protein (AtEBP),an ethylene inducible responsive gene expression,respectively,in Arabidopsis.Plant Cell GCC box DNA-binding protein interacts with an ocs element 10:1391-1406 binding protein.Proc Natl Acad Sci USA 94:5961-5966 Medina J,Bargues M,Terol J,Perez-Alonso M,Salinas J(1999)The Chinnusamy V,Ohta M,Kanrar S,Lee BH,Hong XH,Agarwal M, Arabidopsis CBF gene family is composed of three genes Zhu JK (2003)ICEl:a regulator of cold-induced transcriptome encoding AP2 domain-containing proteins whose expression is and freezing tolerance in Arabidopsis.Genes Dev 17:1043-1054 regulated by low temperature but not by abscisic acid or Chinnusamy V.Zhu JH,Zhu JK(2007)Cold stress regulation of gene dehydration.Plant Physiol 119:463-470 expression in plants.Trends Plant Sci 12:444-451 Meng SS,Dane F,Si Y,Ebel R,Zhang CK (2008)Gene expression Chinnusamy V,Zhu JK,Sunkar R(2010)Gene regulation during cold analysis of cold treated versus cold acclimated Poncirus stress acclimation in plants.Methods Mol Biol 639:39-55 trifoliata.Euphytica 164(1):209-219 Fowler DB.Limin AE,Wang S.Ward RW (1996)Relationship Meshi T.Iwabuchi M (1995)Plant transcription factors.Plant Cell between low-temperature tolerance and vernalization response in Physiol36:1405-1420 wheat and rye.Can J Plant Sci 76:37-42 Riechmann JL.Meyerowitz EM (1997)Domain proteins in plant Gao MJ.Allard G.Byass L.Flanagan AM.Singh J(2002)Regulation development.Biol Chem 10:1079-1101 and characterization of four CBF transcription factors from Stockinger EJ,Gilmour SJ,Thomashow MF(1997)Arabidopsis thaliana Brassica napus.Plant Mol Biol 49:459-471 CBFI encodes an AP2 domain-containing transcriptional activator Gilmour SJ,Zarka DG,Stockinger EJ,Salazar MP.Houghton JM. that binds to the C-repeat/DRE,a cis-acting DNA regulatory Thomashow MF (1998)Low temperature regulation of the element that stimulates transcription in response to low temperature Arabidopsis CBF family of AP2 transcriptional activators as an and water deficit.Proc Natl Acad Sci USA 94:1035-1040 early step in cold-induced COR gene expression.Plant J 16:433- Thomashow MF (1999)Plant cold acclimation:freezing tolerance 442 genes and regulatory mechanisms.Annu Rev Plant Physiol Plant Gilmour SJ,Sebolt AM,Salazar MP,Everard JD,Thomashow MF Mol Biol 50:571-599 (2000)Overexpression of the Arabidopsis CBF3 transcriptional Thomashow MF(2001)So what's new in the field of plant cold activator mimics multiple biochemical changes associated with acclimation？Lots！Plant Physiol125:89-93 cold acclimation.Plant Physiol 124:1854-1865 Wang XL,Sun XQ,Liu SX,Liu L,Liu XJ,Sun XF,Tang KX(2005) Goulas E,Dily FL,Ozouf J,Ourry A (2003)Effects of a cold Molecular cloning and characterization of a novel ice gene from treatment of the root system on white clover(Trifolium repens L.) Capsella bursa-pastoris.Mol Biol 39:18-25 morphogenesis and nitrogen reserve accumulation.J Plant Wang L,Li XW,Zhao Q.Jing SL,Chen SF,Yuan HY (2009) Physiol160:893-902 Identification of genes induced in response to low-temperature Guiltinan MJ,Marcotte WR,Quatrano RS (1990)A plant leucine treatment in tea leaves.Plant Mol Biol Rep 27:257-265 zipper protein that recognizes an abscisic acid response element. Williams ME,Foster R,Chua NH(1992)Sequences flanking the Science250:267-271 hexameric G-box core CACGTG affect the specificity of protein Huang H,Tudor M,Su T,Zhang Y,Hu Y,Ma H(1996)DNA binding binding.Plant Cell 4:485-496 properties of two Arabidopsis MADS domain proteins:binding Wilson CL,Hubbard SJ,Doig AJ(2002)A critical assessment of the consensus and dimer formation.Plant Cell 8:81-94 secondary structure a-helices and their termini in proteins Jaglo-Ottosen KR,Gilmour SJ,Zarka DG,Schabenberger O, Protein Eng 15(7):545-554 Thomashow MF (1998)Arabidopsis CBFI overexpression induces Yamaguchi-Shinozaki K,Shinozaki K(1994)A novel cis-acting element COR genes and enhances freezing tolerance.Science 280:104-106 in an Arabidopsis gene is involved in responsiveness to drought. Jaglo-Ottosen KR,Kleff S,Amundsen KL,Zhang X,Haake V,Zhang low-temperature,or high-salt stress.Plant Cell 6:251-264 JZ,Deits T.Thomashow MF (2001)Components of the Yang TW,Zhang LJ,Zhang TG,Zhang H,Xu SJ,An LZ (2005) Arabidopsis C-repeat/dehydration-responsive element binding Transcriptional regulation network of cold-responsive genes in factor cold-response pathway are conserved in Brassica napus higher plants.Plant Sci 169:987-995 and other plant species.Plant Physiol 127:910-917 Zhou N,Robinson SJ,Huebert T,Bate NJ,Parkin IAP (2007) Jiang FL,Hou XL,Shi GJ,Cui XM(2007a)Cloning and characterization Comparative genome organization reveals a single copy of CBF in of full length cDNA of BrCBF gene from Brassica campestris ssp. the freezing tolerant crucifer Thlaspi arvense.Plant Mol Biol chinensis.J Nanjing Agri Uni 30:18-22 (in chinensis) 65:693-705 么Springer(Chinnusamy et al. 2010). Does non-heading Chinese cabbage have pre-mRNA processing in cold acclimation? Further studies also need to be carried out. Acknowledgements This research was partially supported by the Natural Science Foundation of Jiangsu Province (BK2009311) and National Science and Technology Support Program (2009BADB8B03-1). References Altus NN, Uemura M, Steponkus PL, Gilmour SJ, Lin C, Thomashow MF (1996) Constiutive expression of the cold-regulated Arabidopsis thaliana COR15a gene affects both chloroplast and protoplast freezing tolerance. Plant Biol 93:13404–13409 Büttner M, Singh KB (1997) Arabidopsis thaliana ethylene respon￾sive element binding protein (AtEBP), an ethylene inducible, GCC box DNA-binding protein interacts with an ocs element binding protein. Proc Natl Acad Sci USA 94:5961–5966 Chinnusamy V, Ohta M, Kanrar S, Lee BH, Hong XH, Agarwal M, Zhu JK (2003) ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes Dev 17:1043–1054 Chinnusamy V, Zhu JH, Zhu JK (2007) Cold stress regulation of gene expression in plants. Trends Plant Sci 12:444–451 Chinnusamy V, Zhu JK, Sunkar R (2010) Gene regulation during cold stress acclimation in plants. Methods Mol Biol 639:39–55 Fowler DB, Limin AE, Wang S, Ward RW (1996) Relationship between low-temperature tolerance and vernalization response in wheat and rye. Can J Plant Sci 76:37–42 Gao MJ, Allard G, Byass L, Flanagan AM, Singh J (2002) Regulation and characterization of four CBF transcription factors from Brassica napus. Plant Mol Biol 49:459–471 Gilmour SJ, Zarka DG, Stockinger EJ, Salazar MP, Houghton JM, Thomashow MF (1998) Low temperature regulation of the Arabidopsis CBF family of AP2 transcriptional activators as an early step in cold-induced COR gene expression. Plant J 16:433– 442 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 Goulas E, Dily FL, Ozouf J, Ourry A (2003) Effects of a cold treatment of the root system on white clover (Trifolium repens L.) morphogenesis and nitrogen reserve accumulation. J Plant Physiol 160:893–902 Guiltinan MJ, Marcotte WR, Quatrano RS (1990) A plant leucine zipper protein that recognizes an abscisic acid response element. Science 250:267–271 Huang H, Tudor M, Su T, Zhang Y, Hu Y, Ma H (1996) DNA binding properties of two Arabidopsis MADS domain proteins: binding consensus and dimer formation. Plant Cell 8:81–94 Jaglo-Ottosen KR, Gilmour SJ, Zarka DG, Schabenberger O, Thomashow MF (1998) Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance. Science 280:104–106 Jaglo-Ottosen KR, Kleff S, Amundsen KL, Zhang X, Haake V, Zhang JZ, Deits T, Thomashow MF (2001) Components of the Arabidopsis C-repeat/dehydration-responsive element binding factor cold-response pathway are conserved in Brassica napus and other plant species. Plant Physiol 127:910–917 Jiang FL, Hou XL, Shi GJ, Cui XM (2007a) Cloning and characterization of full length cDNA of BrCBF gene from Brassica campestris ssp. chinensis. J Nanjing Agri Uni 30:18–22 (in chinensis) Jiang FL, Hou XL, Shi GJ, Cui XM (2007b) Cloning and characterization of full length cDNA of BrCOR14 gene from Brassica campestris ssp. chinensis. Jiangsu J of Agr Sci 23:34– 38 (in Chinese) Jiang FL, Hou XL, Shi GJ, Cui XM (2007c) Cloning and characterization of full length cDNA of BrLOS2 gene from Brassica campestris ssp. chinensis. J Nanjing Agri Uni 30:27–32 (in Chinese) Kasuga M, Liu Q, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (1999) Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nat Biotechnol 17:287–291 Lee B, Henderson DA, Zhu JK (2005) The arabidopsis cold￾responsive transcriptome and its regulation by ICE1. Plant Cell 17:3155–3175 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 low-temperature￾responsive gene expression, respectively, in Arabidopsis. Plant Cell 10:1391–1406 Medina J, Bargues M, Terol J, Pérez-Alonso M, Salinas J (1999) The Arabidopsis CBF gene family is composed of three genes encoding AP2 domain-containing proteins whose expression is regulated by low temperature but not by abscisic acid or dehydration. Plant Physiol 119:463–470 Meng SS, Dane F, Si Y, Ebel R, Zhang CK (2008) Gene expression analysis of cold treated versus cold acclimated Poncirus trifoliata. Euphytica 164(1):209–219 Meshi T, Iwabuchi M (1995) Plant transcription factors. Plant Cell Physiol 36:1405–1420 Riechmann JL, Meyerowitz EM (1997) Domain proteins in plant development. Biol Chem 10:1079–1101 Stockinger EJ, Gilmour SJ, Thomashow MF (1997) Arabidopsis thaliana CBF1 encodes an AP2 domain-containing transcriptional activator that binds to the C-repeat/DRE, a cis-acting DNA regulatory element that stimulates transcription in response to low temperature and water deficit. Proc Natl Acad Sci USA 94:1035–1040 Thomashow MF (1999) Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu Rev Plant Physiol Plant Mol Biol 50:571–599 Thomashow MF (2001) So what’s new in the field of plant cold acclimation? Lots! Plant Physiol 125:89–93 Wang XL, Sun XQ, Liu SX, Liu L, Liu XJ, Sun XF, Tang KX (2005) Molecular cloning and characterization of a novel ice gene from Capsella bursa-pastoris. Mol Biol 39:18–25 Wang L, Li XW, Zhao Q, Jing SL, Chen SF, Yuan HY (2009) Identification of genes induced in response to low-temperature treatment in tea leaves. Plant Mol Biol Rep 27:257–265 Williams ME, Foster R, Chua NH (1992) Sequences flanking the hexameric G-box core CACGTG affect the specificity of protein binding. Plant Cell 4:485–496 Wilson CL, Hubbard SJ, Doig AJ (2002) A critical assessment of the secondary structure α-helices and their termini in proteins. Protein Eng 15(7):545–554 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 Yang TW, Zhang LJ, Zhang TG, Zhang H, Xu SJ, An LZ (2005) Transcriptional regulation network of cold-responsive genes in higher plants. Plant Sci 169:987–995 Zhou N, Robinson SJ, Huebert T, Bate NJ, Parkin IAP (2007) Comparative genome organization reveals a single copy of CBF in the freezing tolerant crucifer Thlaspi arvense. Plant Mol Biol 65:693–705 532 Plant Mol Biol Rep (2011) 29:525–532