外源脱水应答转录因子CBF4基因转化玉米的研究
摘要
玉米是世界上三大粮食作物之一,也是重要的饲料作物,在世界农业生产中占有相当重要的地位。在我国,玉米的总产已经超过小麦,成为第二大粮食作物。但是,我国每年约有50%以上的玉米种植在干旱和半干旱地区。干旱是限制我国玉米生产发展和产量提高的第一要素。因此,培育抗旱玉米品种非常重要。以杂种优势利用为核心的传统育种方法在提高玉米产量上已取得了巨大成功。然而,玉米的抗旱性状属于数量性状,由于传统育种方法的局限性,已经不能满足人们越来越高的需求。近年来,利用转基因技术提高玉米抗旱能力的研究受到世界各国科学家的广泛关注。
目前,利用基因工程技术培育抗旱品种主要有以下几种策略,包括转渗透调节物质合成关键酶基因、转活性氧清除酶类基因、转抗旱基因和转编码转录因子的基因等。转录因子由于能激活下游一系列抗逆基因的表达而逐渐受到人们的重视,转转录因子抗旱基因工程与转单个抗旱基因相比具有更大的优越性。为了提高玉米的抗旱能力,加速我国玉米抗旱育种的进程,本研究克隆了拟南芥脱水应答转录因子CBF4基因并将其转化到玉米中去,以期得到抗旱能力有所提高的转基因玉米株系,主要取得以下进展:
1)克隆了拟南芥CBF4基因和逆境诱导表达基因rd29A的启动子,以草甘膦为筛选标记基因,构建了rd29A启动子驱动下的CBF4基因逆境诱导表达载体pBAC146;
2)分别用基因枪法和花粉管通道法将pBAC146载体转化到玉米优良自交系中,其中基因枪转化法获得36株转化植株,花粉管通道法收获了1270粒转化种子;
3)基因枪法获得的T_0代转基因植株经PCR检测大部分呈阳性,南繁后4株转基因植株成功结实,对T_1代转基因株系的PCR、PCR-Southern、Southern杂交检测表明,3个转基因株系成功整合了外源目的基因CBF4;
4)为检测所获得的转基因植株的抗旱能力,将T_1代转基因植株收获的种子(T_2代)种植到温室花盆中进行人工抗旱处理后,对其抗旱相关生理指标,包括相对含水量,叶绿素含量和脯氨酸含量等进行检测,各项指标测定结果均表明,转基因植株的抗旱能力相比对照有所提高;
5)由于花粉管通道法获得的种子较多,为进行快速有效地筛选,进行了草甘膦预实验,筛选出了合适的草甘膦浓度,为下一步阳性植株的筛选奠定了基础。
本研究表明,利用脱水应答转录因子CBF4基因在转基因玉米中的诱导表达,可以提高玉米的抗旱能力,我们的工作为进一步为利用CBF4基因进行玉米抗旱育种奠定了基础。
Maize,one of the three important cereal crops in the world,plays a very important role in food and animal feed production.As its whole production surpassed wheat,maize has become the second largest cereal crops in china.More than fifty percent of maize planted in arid and semiarid region every year.And through the whole growth stage of maize,different degrees of damage to its growth might be caused by drought and less of water,which are the main factors restricting maize production.So,it is very important to breed drought resistant maize.Traditional breeding way using heterosis as a main method has made a great success on improvement of maize production.As quantitative character of drought resistance, traditional breeding has too many limitations to satisfy the needing of breeders in the future. In recent years,to improve drought resistance of maize by transgenic method has got great attention in many countries.
There are several kinds of gene we can use in breeding stress resistant plant,Genes for osmotic regulation substance synthesis,for example,genes coding for enzymes for active oxygen clearing,genes coding for proteins induced by abiotics or genes acts in the stress signal transduction pathway.In order to gain high drought resistance maize and speed up the process of drought resistance breeding,the dehydration responsive transcriptional factor CBF4 gene from Arabidopsis thaliana was cloned and transformed into maize with biolistic bombardment.The main results are as follows:
1) The CBF4 gene and the promoter of rd29A gene,a preceding drought responsive gene, were amplified from Arabidopsis thaliana by PCR.Using the EPSP gene as the selective marker,pBAC146,the expression vector of CBF4 gene drived by the promoter of rd29A gene was constructed.
2) Biolistic bombardment and pollen-tube pathway were used to transform the pBAC146 vector into excellent maize inbred lines.Thirty six transformation plants transformed by Biolistic bombardment were obtained and 1270 seeds were harvested through the pollen-tube pathway.
3) PCR analysis suggested that the transgene exist in most of the 36 transformation plants, among which four plants have successfully gotten seeds.PCR,PCR-Southern and Southern analysis proved that the alien CBF4 gene had been integrated into the genome of 3 transgenic lines.
4) To test the drought resistence ability of the transgenic lines we got,seeds of T1 generation (T_2 generation) were planted into garden pot in greenhouse and treated by no watering. The relative water content,proline content and chlorophyll content analysis showed that transgenic lines had higher drought resistant ability than non-transgenic plants.
5) To select those seeds obtained by pollen tube pathway quickly and effectively,normal maize seeds were treated by different content of glyphosate.The suitable glyphosate content for distinguishing transgenic and non-transgenic plants was found.
Thisr study suggested that induced expression of the CBF4 gene in transgenic maize could improve the drought resistance of transgenic maize,and which will pave a way for using CBF4 gene in maize drought resistant breeding.
目前,利用基因工程技术培育抗旱品种主要有以下几种策略,包括转渗透调节物质合成关键酶基因、转活性氧清除酶类基因、转抗旱基因和转编码转录因子的基因等。转录因子由于能激活下游一系列抗逆基因的表达而逐渐受到人们的重视,转转录因子抗旱基因工程与转单个抗旱基因相比具有更大的优越性。为了提高玉米的抗旱能力,加速我国玉米抗旱育种的进程,本研究克隆了拟南芥脱水应答转录因子CBF4基因并将其转化到玉米中去,以期得到抗旱能力有所提高的转基因玉米株系,主要取得以下进展:
1)克隆了拟南芥CBF4基因和逆境诱导表达基因rd29A的启动子,以草甘膦为筛选标记基因,构建了rd29A启动子驱动下的CBF4基因逆境诱导表达载体pBAC146;
2)分别用基因枪法和花粉管通道法将pBAC146载体转化到玉米优良自交系中,其中基因枪转化法获得36株转化植株,花粉管通道法收获了1270粒转化种子;
3)基因枪法获得的T_0代转基因植株经PCR检测大部分呈阳性,南繁后4株转基因植株成功结实,对T_1代转基因株系的PCR、PCR-Southern、Southern杂交检测表明,3个转基因株系成功整合了外源目的基因CBF4;
4)为检测所获得的转基因植株的抗旱能力,将T_1代转基因植株收获的种子(T_2代)种植到温室花盆中进行人工抗旱处理后,对其抗旱相关生理指标,包括相对含水量,叶绿素含量和脯氨酸含量等进行检测,各项指标测定结果均表明,转基因植株的抗旱能力相比对照有所提高;
5)由于花粉管通道法获得的种子较多,为进行快速有效地筛选,进行了草甘膦预实验,筛选出了合适的草甘膦浓度,为下一步阳性植株的筛选奠定了基础。
本研究表明,利用脱水应答转录因子CBF4基因在转基因玉米中的诱导表达,可以提高玉米的抗旱能力,我们的工作为进一步为利用CBF4基因进行玉米抗旱育种奠定了基础。
Maize,one of the three important cereal crops in the world,plays a very important role in food and animal feed production.As its whole production surpassed wheat,maize has become the second largest cereal crops in china.More than fifty percent of maize planted in arid and semiarid region every year.And through the whole growth stage of maize,different degrees of damage to its growth might be caused by drought and less of water,which are the main factors restricting maize production.So,it is very important to breed drought resistant maize.Traditional breeding way using heterosis as a main method has made a great success on improvement of maize production.As quantitative character of drought resistance, traditional breeding has too many limitations to satisfy the needing of breeders in the future. In recent years,to improve drought resistance of maize by transgenic method has got great attention in many countries.
There are several kinds of gene we can use in breeding stress resistant plant,Genes for osmotic regulation substance synthesis,for example,genes coding for enzymes for active oxygen clearing,genes coding for proteins induced by abiotics or genes acts in the stress signal transduction pathway.In order to gain high drought resistance maize and speed up the process of drought resistance breeding,the dehydration responsive transcriptional factor CBF4 gene from Arabidopsis thaliana was cloned and transformed into maize with biolistic bombardment.The main results are as follows:
1) The CBF4 gene and the promoter of rd29A gene,a preceding drought responsive gene, were amplified from Arabidopsis thaliana by PCR.Using the EPSP gene as the selective marker,pBAC146,the expression vector of CBF4 gene drived by the promoter of rd29A gene was constructed.
2) Biolistic bombardment and pollen-tube pathway were used to transform the pBAC146 vector into excellent maize inbred lines.Thirty six transformation plants transformed by Biolistic bombardment were obtained and 1270 seeds were harvested through the pollen-tube pathway.
3) PCR analysis suggested that the transgene exist in most of the 36 transformation plants, among which four plants have successfully gotten seeds.PCR,PCR-Southern and Southern analysis proved that the alien CBF4 gene had been integrated into the genome of 3 transgenic lines.
4) To test the drought resistence ability of the transgenic lines we got,seeds of T1 generation (T_2 generation) were planted into garden pot in greenhouse and treated by no watering. The relative water content,proline content and chlorophyll content analysis showed that transgenic lines had higher drought resistant ability than non-transgenic plants.
5) To select those seeds obtained by pollen tube pathway quickly and effectively,normal maize seeds were treated by different content of glyphosate.The suitable glyphosate content for distinguishing transgenic and non-transgenic plants was found.
Thisr study suggested that induced expression of the CBF4 gene in transgenic maize could improve the drought resistance of transgenic maize,and which will pave a way for using CBF4 gene in maize drought resistant breeding.
引文
Abebe T,Guenzi A C,Martin B,et al.Tolerance of mannitol-accumulating transgenic wheat to water stress and salinity.Plant Physiol,2003,131(4):1748-1755.
Alia,H H,Sakamoto A,Murata N.Enhancement of the tolerance of Arabidopsis to high temperatures by genetic engineering of the synthesis of glycinebetaine.Plant J.1998,16(2):155-61.
Apel K,Hirt H..Reactive oxygen species:metabolism,oxidative stress,and signal transduction.Annu Rev Plant Biol,2004,55:373-99.
Bahieldin A,Mahfouz H T,Eissa H F,et al.Field evaluation of transgenic wheat plants stably expressing the HVA1 gene for drought tolerance.Physiol Plant,2005,123(4):421-427.
Baker S S,Wilhelm K S,Thomashow M F.The 5'-region of Arabldopsis thaliana cor15a has cis-acting elements that confer cold-,drought- and ABA-regulated gene expression.Plant Mol Biol,1994,24:701-713.
Bender J,Fink G R.A Myb homologue,ATR1,activates tryptophan gene expression in Arabidopsis.PNAS,1998,95(10):5655-5660.
Camp W V,Capiau K,Montagu M V,et al.Enhancement of oxidative stress tolerance in transgenic tobacco plants overproducing Fe-superoxide dismutase in chloroplasts.Plant Physiol,1996,112(4):1703-1714.
Chen B J,Wang Y,Hu Y L,et al.Cloning and characterization of a drought inducible MYB gene from Boea crassifolia.Plant Sci,2005,168(2):493-500.
Chen J Q,Meng X P,Zhang Y,et al.Over-expression of OsDREB genes lead to enhanced drought tolerance in rice.Biotechnol Lett,2008,30(12):2191-2198.
Chen M,Xu Z,Xia L,et al.Cold-induced modulation and functional analyses of the DRE-binding transcription factor gene,GmDREB3,in soybean(Glycine max L.).J Exp Bot,2009,60(1):121-135.
Cheng Z,Targolli J,Huang X Q,et al.Wheat LEA genes,PMA80 and PMA1959,enhance dehydration tolerance of transgenic rice(Oryza sativa L.).Mol Breed,2002,10:71-82.
Cho E K,Hong C B.Over-expression of tobacco NtHSP70-l contributes to drought-stress tolerance in plants.Plant Cel I Rep,2006,25:349-358.
Cominelli E,Galbiati M,Vavasseur A,et al.A guard cell specific MYB transcription factor regulates stomatal movements and plant drought tolerance.Curr Biol,2005,15(13):1196-2000.
Could J,Devery M,Hasegawa 0.Transformation of Zea mays L.using Agrobacterium tumefaciens and the shoot apex.Plant Physiol,1991,95:426-434.
Dai X Y,Xu Y Y,Ma Q B,et al.Overexpression of an R1R2R3 MYB gene,OsMYB3R-2,increases tolerance to freezing,drought,and salt stress in transgenic Arabidopsis.Plant Physiol,2007,143(4):1739-1751.
Delauney A J,Hu C A,Kishor P B,et al.Cloning of ornithine delta-aminotransferase cDNA from Vigna aconitifolia by trans-complementation in Escherichia coli and regulation of proline biosynthesis.J Biol Chem,1993,268(25):18673-18678.
Dubouzet J G,Sakuma Y,Ito Y,Kasuga M,et al.OsDREB genes in rice,Oryza sativa L.,encode transcription activators that function in drought-,high-salt- and cold-responsive gene expression.Plant J,2003,33(4):751-763.
Eulgem T,Rushton PJ,Robatzek S,et al.The WRKY super family of plant transcription factors.Trends Plant Sci,2000,5(5):199-206.
Frame B R,Shou H,Chikwamba R K,et al.Agrobacterium tumefaciens-mediated transformation of maize embryos using a standard binary vector system.Plant Physiol,2002,129(1):13-22.
Fromm M E,Taylor L P,Valbot V.Stable transformation of maize after gene transfer by eletroporation.Nature,1986,319(6056):791-793.
Fujita M,Fujita Y,Maruyama K,et al.A dehydration-induced NAC protein,RD26,is involved in a novel ABA-dependent stress-signaling pathway.The Plant Journal,2004,39:863-876.
Garg A K,Kim J K,Owens T G,et al.Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses.Proc Natl Acad Sci USA,2002,99(25):15898-15903.
Gilmour S J,Sebolt A M,Salazar M P,et al.Over expression of the Arabidopsis CBF3 transcriptional activator mimics multiple biochemical changes associated with cold acclimation.Plant Physiol,2000,124:1854-1865.
Gilmour S J,Zarka D G,Stockinger E J,et al.Low temperature regulation of the Arabidopsis CBF family of AP2 transcriptional activators as an early step in cold-induced COR gene expression.Plant J,1998,16(4):433-442.
Gupta A S,Heinen J L,Holaday A S,et al.Increased resistance to oxidative stress in transgenic plants that overexpress chloroplastic Cu/Zn superoxide dismutase.Proc Natl Acad Sci USA,1993,90(4):1629-1633.
Gutha L R,Reddy A R.Rice DREB1B promoter shows distinct stress-specific responses,and the overexpression of cDNA in tobacco confers improved abiotic and biotic stress tolerance.Plant Mol Biol,2008,68(6):533-555.
Haake V,Cook D,Riechmann J L,et al.Transcription factor CBF4 is a regulator of drought adaptation in Arabidopsis.Plant physiology,2002,130(2):639-648.
Hern(?)ndez G,Ram(?)rez M,Vald(?)s-L(?)pez O,et al.Phosphorus stress in common bean:root transcript and metabolic responses.Plant Physiol,2007,144(2):752-767.
Hiei Y,Ohta S,Komari T,et al.Efficient Transformation of Rice(Oryza Sativa L.)Mediated by Agrobacterium and Sequence Analysis of the Boundaries of the T-DNA.Plant Journal,1994,6(2):271-282.
Hohmann S,Bell W,Neves M J,et al.Evidence for trehalose-6-phosphate-dependent and -independent mechanisms in the control of sugar influx into yeast glycolysis.Mol Microbiol,1996,20(5):981-991.
Holmstrom K O,Welin B,Mandal A,et al.Production of the Escherichia coli betaine-aldehyde dehydrogenase,an enzyme required for the synthesis of the osmoprotectant glycine betaine,in transgenic plants.Plant J,1994,6(5):749-58.
Hsieh T H,Lee J T,Charng Y Y,et al.Tomato plants ectopicaly expressing Arabidopsis CBF1 show enhanced resistance to water deficit stress.Plant Physiology,2002,130(2):618-626.
Ishida Y,Saito H,Ohta S,et al.High efficiency transformation of maize(Zea mays L.)mediated by Agrobacterium tumefaciens.NatBiotechnol,1996,14(6):745-750.
Ishiguro S,Nakamura K.Characterization of a cDNA encoding a novel DNA-binding protein,SPFI,that recognizes SP8 sequences in the 5'upstream regions of genes coding for sporamin and 6-amylase from sweet potato.Mol Gen Genet,1994,244(6):563-571.
Jaglo-Ottosen K R,Gilmour S J,Zarka D G,Schabenberger O,et al.Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance.Science,1998,280(5360):104-106.
Jeanneau M,Gerentes D,Foueillassar X,et al.Improvement of drought tolerance in maize:towards the functional validation of the Zm-Asrl gene and increase of water use efficiency by over-expressing C4-PEPC.Biochimie,2002,84(11):1127-1135.
Jiang C,lu B,Singh J.Requirement of a CCGAC cis-acting element for cold induction of the BN115 gene from,winter Brassica napus.Plant Mol.Biol,1996,30(3):679-684.
Jiang Y,Deyholos M K.Functional characterization of Arabidopsis NaCl-inducible WRKY25 and WRKY33 transcription factors in abiotic stresses.Plant Mol Biol,2009,69(1-2):91-105.
Jin H,Martin C.Multifunctionality and diversity within the plant MYB-gene family.Plant Mol Biol,1999,41(5):577-585.
Kang J Y,Choi H I,Im M Y,et al.Arabidopsis basic leucine zipper proteins that mediate stress-responsive abscisic acid signaling.Plant Cell,2002,14(2):343-357.
Katiyar-Agarwal S,Agarwal M,Grover A.Heat-tolerant basmati rice engineered by over-expression of hsp101.Plant Mol Biol,2003,51(5):677-686.
Kenward K D,Altschuler M,Hildebrand D,et al.Accumulation of type I fish antifreeze protein in transgenic tobacco is cold-spesific.Plant Mol Biol,1993,23(2):377-385.
Kishor PBK,Hong Z,Miao G H,et al.Over expression of[delta]-Pyrroline-5 -Carboxylate Synthetase Increases Proline Production and Confers Osmotolerance in Transgenic Plants.Plant Physiol,1995,108(4):1387-1394.
Klein T M,Kornstein L,Sanford J C,et al.Transformation of Maize Cells by Particle Bombardment.Plant Physiology,1989,91(1):440-444.
Koziel M G,Beland G L,Bowman C,et al.Field performence of elite transgenic maize plant expressing all insecticidal protein derived from Bacillus thuringiensis.Bio/Technolgy,1993,11:194-200.
Lea U S,Slimestad R,Smedvig P,et al.Nitrogen deficiency enhances expression of specific MYB and bHLH transcription factors and accumulation of end products in the flavonoid pathway.Planta,2007,225(5):1245-1253.
Lee S C,Choi H W,Hwang I S,et al.Functional roles of the pepper pathogen-induced bZIP transcription factor,CAbZIPl,in enhanced resistance to pathogen infection and environmental stresses.Planta,2006,224(5):1209-1225.
Li S,Fu Q,Huang W,Yu D.Functional analysis of an Arabidopsis transcription factor WRKY25 in heat stress.Plant Cell Rep,2009,Epub ahead of print.Lipsick J S.One billion years of Myb.Oncogene,1996,13(2):223-235.
Liu Q,Kasuga M,Sakuma Y,et al.Two transcription factors,DREBl 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,1998,10:1391-1406.
Liu Y,Wang G,Liu J,et al.Transfer of E.coli gutD gene into maize and regeneration of salt-tolerant transgenic plants.Sci China C Life Sci,1999,42(1):90-95.
Lu G,Gao C,Zheng X,et al.Identification of OsbZIP72 as a positive regulator of ABA response and drought tolerance in rice.Planta,2009,229(3):605-615.
Lung J,Giraudat J.Abscisic acid signal transduction.Annu Rev.Plant Physiol Plant Mol Biol,1998,49:199-222.
Lupotto E,Reali A,Oassera S,et al.Maize elite inbred lines are susceptible to Agrobacterium tumefaciens-mediated transformation.Maydica,1999,44(3):211-218.
Martin C,Paz-Ares J.MYB transcription factors in plants.Trends Genet,1997,13(2):67-73.
Medina J,Bargues M,Terol J,et al.The Arabidopsis CBF gene family composed of three gene encoding AP2 domain-containing proteins whose expression is regulated by low temperature but not by abscisic acid or dehydration.Plant Physiol,1999,119:463-469.
Miyake K,Ito T,Sends M.Isolation of a subfamily of genes for R2R3-MYB transcription factors showing up-regulated expression under nitrogen nutrient-limited conditions,Plant Mol Biol,2003,53(1-2):237-245.
Moffat A S.Finding New Ways to Protect Drought-Stricken Plants.Science,2002,296(5571):1226-1229.
Moghaieb REA,Tanaka N.Sancoka H,et al.Expression of betaine aldehyde dehydrogenase gene in transgenic tomato hairy roots leads to the accumulation of glycine betaine and contributes to the maintenance of the osmotic potential under salt stress.Soil Sci Plant Nutr,2000,46(4):873-883.
Nanjo T,Kobayashi M,Yoshiba Y,et al.Antisense suppression of proline degradation improves tolerance to freezing and salinity in Arabidopsis thaliana.FEBS Lett,1999,461(3):205-210.
Ohnishi T,Sugahara S,Yamada T,et al.OsNAC6,a member of the NAC gene family,is induced by various stresses in rice.Genes Genet Syst,2005,80:135-139.
Ouellet F,Vazquez-Tello A,Sarhan F.The wheat Wcs120 promoter is cold-inducible in both monocotyledonous and dicotyledonous species.FEBS Letters,1998,423(3):324-328.
Pardo J M,Reddy M P,Yang S,et al.Stress signaling through Ca2+/calmodulin-dependent protein phosphatase calcineurin mediates salt adaptation in plants.Proc Natl Acad Sci USA,1998,95(16):9681-9686.
Park E J,Jeknic Z,Sakamoto A,et al.Genetic engineering of glycinebetaine synthesis in tomato protects seeds,plants,and flowers from chilling damage.Plant J,2004,40(4):474-487.
Perl A,Perl-Treves R,Galili S,et al.Enhanced oxidative stress defense in transgenic potato expressing tomato Cu/Zn superoxide dismutases.Theor Appl Genet,1993,85(5):568-576.
Qin F,Kakimoto M,Sakuma Y,et al.Regulation and functional analysis of ZmDREB2A in response to drought and heat stresses in Zea mays L.Plant J,2007,50(1):54-69.
Qin F,Sakuma Y,Li J,et al.Cloning and functional analysis of a novel DREB1/CBF transcription factor involved in cold-responsive gene expression in Zea mays L.Plant Cell Physiol,2004,45(8):1042-1052.
Quan R,Shang M,Zhang H,et al.Engineering of enhanced glycine betaine synthesis improves drought tolerance in maize.Plant Biotechnol J,2004,2(6):477-486.
Rabinowicz P D,Braun E L,Wolfe A D,et al.Maize R2R3 MYB gene:sequence analysis reveals amplification in the higher plants.Genetics,1999,153(1):427-444.
Rhodes C A,Pierce D A,Mettler I J,et al.Genetically transformed maize plants from protoplast.Sciene,1988,240(4849):204-207.
Riechmann J L,Heard J,Martin G,et al.Arabidopsis transcription factors:genome-wide comparative analysis among Eukaryotes.Science,2000,290(5499):2105-2110.
Riechmann J L,Meyerowitz E M.The AP2/EREBP family of plant transcription factors.Biol Chem,1998,379(6):633-646.
Rubio M C,Gonzalez E M,Minchin F R,et al.Effects of water stress on antioxidant enzymes of leaves and nodules of transgenic alfalfa overexpressing superoxide dismutases.Physiol Plant,2002,115(4):531-540.
Saijo Y,Hata S,Kyozuka J,et al.Over-expression of a single Ca~(2+)-dependent protein kinase confers both cold and salt/drought tolerance on rice plants.Plant J,2000,23(3):319-327.
Sakuma Y,Liu Q,Dubouzet J G,et al.DNA-binding specificity of the ERF domain of Arabidopsis DREBs.transcription factors involved in dehydration and cold-inducible gene expression.Biochem Biophys Res Commun,2002,290(3):998-1009.
Sanmiya K,Suzuki K,Egawa Y,Shono M.Mitochondrial small heat-shock protein enhances thermotolerance in tobacco plants.FEBS Lett,2004,557(1-3):265-268.
Sayari AH,Bouzid R G,Bidani A,et al.Overexpression of Al-pyrroline-5-carboxylate synthetase increases proline production and confers salt tolerance in transgenic potato plants.2005,169(4):746-752.
Selth L A,Dogra S C,Rasheed M S,et al.A NAC domain protein interacts with tomato leaf curl virus replication accessory protein and enhances viral replication.Plant Cell,2005,17(1):311-325.
Shen B,Jensen R G,Bohnert H J.Increased resistance to oxidative stress in transgenic plants by targeting mannitol biosynthesis to chloroplasts.Plant Physiol.1997,113(4):1177-83.
Shinwari Z K,Nakashima K,Miura S,et al.An Arabidopsis gene family encoding DRE/CRT binding proteins involved in low-temperature-responsive gene expression.Biochem Biophys Res Commun,1998,250(1):161-170.
Shou H,Bordallo P and Wang K.Expression of the Nicotiana protein kinase(NPK1)enhanced drought tolerance in transgenic maize.J Exp Bot,2004,55(399):1013-1019.
Sivamani E,Bahieldin A,Wraith J M,et al.Improved biomass productivity and water use efficiency under water deficit conditions in transgenic wheat constitutively expressing the barley HVA1 gene.Plant Sci,2000,155(1):1-9.
Slooten L,Capiau K,Camp W V,et al.Factors affecting the enhancement of oxidative stress tolerance in transgenic tobacco overexpressing manganese superoxide dismutase in the chloroplasts.Plant Physiol,1995,107(3):737-775.
Stockinger E J,Gilmour S J,Thomashow M F.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,1997,94(3):1035-1040.
Stockinger E J,Mao Y,Regier M K,et al.Transcriptional adaptor And histon acetyltransferase proteins in Arabidopsis and their interactions with CBF1,a transcriptional adaptor involved in cold-regulated gene expression.Nucleic Acids Res,2001,29(7):1524-1533.
Stracke R,Werber M,Weisshaar B.The R2R3-MYB gene family in Arabidopsis thaliana.Curr Opin Plant Biol,2001,4(5):447-456.
Sun W,Van Montagu M,Verbruggen N.Small heat shock proteins and stress tolerance in plants.Biochim Biophys Acta,2002,1577(1):1-9.
Tang W,Peng X,Newton R J.Enhanced tolerance to salt stress in transgenic loblolly pine simultaneously expressing two genes encoding mannitol-1-phosphate dehydrogenase and glucitol-6-phosphate dehydrogenase.Plant Physiol Biochem,2005,43(2):139-46.
Tran L S,Nakashima K,Sakuma Y,et al.Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 promoter.Plant Cell,2004,6(9):2481-2498.
Tran LS,Nakashima K,Sakuma Y,et al.Isolation and functional analysis of Ambidopsis stress-inducible NAC transcription factors that bind to a drought responsive Cis-element in the EARLY RESPONSIVE TO DEHYDRATION STRESS promoter.Plant Cell,2004,16(9):2481-2498.
Tyerman S D,Niemietz C M,Bramley H.Plant aquaporins:multifunctional water and solute channels with expanding roles.Plant Cell Environ,2002,25(2):173-194.
Ulker B,Somssich I E.WRKY transcription factors:from DNA binding towards biological function.Curr Opin Plant Biol,2004,7(5):491-498.
Umezawa T,Fujita M,Fujita Y,et al.Engineering drought tolerance in plants:discovering and tailoring genes to unlock the future.Curr Opin Biotechnol,2006,17(2):113-122.
Umezawa T,Yoshida R,Maruyama K,et al.SRK2C,a SNFl-related protein kinase 2,improves drought tolerance by controlling stress-responsive gene expression in Arabidopsis thaliana.Proc Natl Acad Sci USA,2004,101(49):17306-17311.
Vannini C,Locatelli F,Bracale M,et al.Overexpression of the rice Osmyb4 gene increases chilling and freezing tolerance of Arabidopsis thaliana plants.Plant J,2004,37(11):115-127.
Vendniscolo E C,Schuster I,Pileggi M,et al.Stress-induced synthesis of proline confers tolerance to water deficit in transgenic wheat.J Plant Physiol,2007,164(10):1367-1376.
Verling E.The roles of heat shock proteins in plants.Annu Rev Plant Physiol Mol Biol,1991,42:579-620.Wang C R,Yang A F,Yue G D,et al.Enhanced expression of phospholipase C 1(ZmPLCl)improves drought tolerance in transgenic maize.Planta,2008,227(5):1127-1140.
Wang J W,Yang F P,Chen X Q,et al.Induced expression of DREB transcriptional factor and study on its physiological effects of drought tolerance in transgenic wheat.Acta Genet Sin,2006,33:468-476.
Wang L,Li X,Chen S,Liu G Enhanced drought tolerance in transgenic Leymus chinensis plants with constitutively expressed wheat TaLEA3.Biotechnol Lett,2009,31(2):313-319.
Wang L,Zhao C M,Wang Y J,et al.Overexpression of chloroplast-localized small molecular heat-shock protein enhances chilling tolerance in tomato plant.J Plant Physiol Mol Biol,2005,31(2):167-174.
Wang W,Vinocur B,Shoseyov O,et al.Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response.Trends Plant Sci,2004,9(5):244-52.
Wu X,Shiroto Y,Kishitani S,et al.Enhanced heat and drought tolerance in transgenic rice seedlings overexpressing OsWRKY11 under the control of HSP101 promoter.Plant Cell Rep,2009,28(1):21-30.X
iong L,Schumaker K S,Zhu J K.Cell signaling during cold,drought,and salt stress.Plant Cell,2002,14Suppl:S165-183.
Xu Z S,Ni Z Y,Li Z Y,et al.Isolation and functional characterization of HvDREBI-a gene encoding a dehydration-responsive element binding protein in Hordeum vulgare.J Plant Res,2009,122(1):121-130.
Yamaguchi-Shinozaki K,Shinozaki K.Improving plant drought,salt and freezing tolerance by gene transfer of a single stress-inducible transcription factor.Novartis Found Symp,2001,236:176-186.
Yamaguchi-Shinozaki K,Shinozaki K.Transcriptional regulatory networks in cellular responses and toleranse to dehydration and cold stress.Annu.Rev.Plant Biol,2006,57:781-803.
Yancey P H.Living with water stress:evolution of osmolyto system.Science,1982,217:1214-1222.
Yokotani N,Ichikawa T,Kondou Y,et al.Tolerance to various environmental stresses conferred by the salt-responsive rice gene ONAC063 in transgenic Arabidopsis.Planta,2009,ahead of print.
Zhao Z Y,Gu W N,Cai T,et al.High throughput genetic transformation mediated by Agrobacterium tumefaciens in maize.Mol.Breed,2002,8(4):323-333.
Zhu J K.Salt and drought stress signal transduction in plants.Annu Rev Plant Biol,2002,53:247-73.
丁群星,谢友菊,戴景瑞等,用子房注射法将Bt毒蛋白基因导入玉米的研究.中国科学(B辑),1993,23(7):707-713.
傅桂荣,张初,赵晓祥等.转美州拟鲽抗冻蛋白基因(AFP)番茄的叶片电导率测定.生物技术,1997,7(5):7-8.
高银.植物抗逆机制与基因工程研究进展.内蒙古农业科技,2007,5:75-78.
黄璐,卫志明.农杆菌介导的玉米遗传转化.实验生物学报,1999,32(4):381-389.
李芳,李先文,黎定军.AP2/EREBP转录因子及其在植物抗逆中的作用.江西农业学报,2008,20(1):44-47.
李慧芬,刘翔,曲强等.转抗虫融合基因(cry1Ac3-cpti)玉米(Zea mays L).自然科学进展,2002,12(1):37-41.
李余良,胡建广,苏菁,刘建华.子房注射法将BI基因导入超甜玉米.玉米科学,2005,13(1):41-43.
李跃强,宣维健,盛承发.植物的低温蛋白.生态学报,2004,249(5):1034-1039.
梁峥,马德饮,汤岚等.菠菜甜菜碱醛脱氢酶基因在烟草中的表达.生物工程学报,1997,(13):236-240.
粱慧敏,夏阳,王太明.植物抗寒冻、抗旱、耐盐基因工程研究进展.草业学报,2003,12(3):1-7.
林茂,闫海霞,眭顺照,李名扬.植物CBF转录因子及其在基因工程中的作用.广西农业科学,2008,39(1):21-25.
林士杰,李俊涛,姜静等.转柽柳晚期胚胎富集蛋白基因烟草的耐低温性分析.生物技术通讯,2006,14(4):563-566.
刘蕾,杜海,唐晓风等.MYB转录因子在植物抗逆胁迫中的作用及其分子机理.遗传,2008,30(10):1265-1271.
刘万勃,宋明,张中灵等.抗冻蛋白的研究进展.生物学杂志,2000,17(4):7-8.
刘欣,李云.转录因子与植物抗逆性研究进展.中国农学通报,2006,22(4):61-66.
刘昀,李冉辉,郑易之等.大豆PM2蛋白及其结构域可提高烟草耐盐性.深圳大学学报:理工版,2007,24(1):95-101.
卢东长城,利用共抑制技术获得高直链淀粉含量的玉米新种质.2008,首都师范大学硕士论文.
路子显,常团结,刘翔等.植物碱性亮氨酸拉链(bZIP)蛋白的研究进展(一)--结构、分类、分布和同源性分析.遗传,2001,23:564-570.
孟凤,郁松林,郊强卿等.甜菜碱与植物抗逆性关系之研究进展.中国农学通报,2008,24(4):225-228.
彭运生,刘恩.关于提取叶绿素方法的比较研究.北京农业大学学报,1992,18(3):247-250.
祁永红.大豆DNA直接导入玉米自交系的研究.玉米科学,2000,8(1):34-36.
钱刚,翟旭光,韩兆雪等.西藏青稞IEA3蛋白新抗旱基因的克隆与序列分析.作物学报,2007'33(2):292-296.
权瑞党,尚梅,王兆玉,张举仁.农杆菌介导的雪花莲凝集素基因转入玉米骨干自交系.西北植物学报,2004,24(5):761-767.
荣红颖,外源脱水应答转录因子DREB1B在转基因小麦后代的稳定表达研究.2008,首都师范大学硕士论文.
苏琦,尚宇航,杜密英,杜进民.植物WRKY转录因子研究进展.农业生物技术科学,2007,23(5):94-98.
孙红伟,朱常香,杨崇良等.玉米基因枪转化Bt基因的研究.山东农业科学,2004,3:20-22.
汤章城.逆境条件下植物脯氨酸累积及其可能的意义.植物生理学通讯,1984(1):15-21.
王罡,杜娟,张艳华.用花粉管通道法将Bt杀虫基囚导入玉米自交系的研究.玉米科学,2002,10(1):36-37.
王国莉,郭振飞.植物耐冷性分子机理的研究进展.植物学通报,2003,20(6):671-679.
王国英,杜天兵,张宏等.用基因枪将Bt毒蛋白基因转入玉米及转基因植株再生.中国科学(B辑),1995,25(1):1-7.
王书子,吴少辉,高海涛等.旱地小麦品种筛选鉴定及其形态特征探讨干旱地区农业研究.,2001,19(2):76-80
武明宇,郝楠.玉米抗旱性的研究进展.现代农业科技,2008,9:124-127.
武模戈,袁刘正.多胺与植物抗逆性关系研究进展.安徽农业科学,2008,36(9):3516-3518.
杨淑慎,山仑,郭蔼光等.水通道蛋白与植物抗旱性.干旱地区农业研究,2005,23(6):214-218.
张明生,谢波,谈锋.甘薯可溶性蛋白、叶绿素及ATP含量变化与品种抗旱性关系的研究.中国农业科学,2003,36(1):13-16.
张淑娟.转盐芥CBF1基因提高玉米抗逆性的研究.2007,山东大学硕士论文.
张艳贞,季静,张领兵等.以基因枪法将BT杀虫蛋白基因导入常规玉米自交系的研究.玉米科学,2001,9(4):23-26.
张正斌.作物抗旱节水的生理遗传育种基础.北京:科学出版社,2003.
周宜君,冯金朝,马文文等.吕惠娟.植物抗逆分子机制研究进展.中央民族大学学报(自然科学版),2006,15(2):169-176.
邹杰,陈信波,刘爱玲等.植物热激蛋白与作物非生物抗逆性的改良.植物生理学通讯,2007,43(5):981-985.
Alia,H H,Sakamoto A,Murata N.Enhancement of the tolerance of Arabidopsis to high temperatures by genetic engineering of the synthesis of glycinebetaine.Plant J.1998,16(2):155-61.
Apel K,Hirt H..Reactive oxygen species:metabolism,oxidative stress,and signal transduction.Annu Rev Plant Biol,2004,55:373-99.
Bahieldin A,Mahfouz H T,Eissa H F,et al.Field evaluation of transgenic wheat plants stably expressing the HVA1 gene for drought tolerance.Physiol Plant,2005,123(4):421-427.
Baker S S,Wilhelm K S,Thomashow M F.The 5'-region of Arabldopsis thaliana cor15a has cis-acting elements that confer cold-,drought- and ABA-regulated gene expression.Plant Mol Biol,1994,24:701-713.
Bender J,Fink G R.A Myb homologue,ATR1,activates tryptophan gene expression in Arabidopsis.PNAS,1998,95(10):5655-5660.
Camp W V,Capiau K,Montagu M V,et al.Enhancement of oxidative stress tolerance in transgenic tobacco plants overproducing Fe-superoxide dismutase in chloroplasts.Plant Physiol,1996,112(4):1703-1714.
Chen B J,Wang Y,Hu Y L,et al.Cloning and characterization of a drought inducible MYB gene from Boea crassifolia.Plant Sci,2005,168(2):493-500.
Chen J Q,Meng X P,Zhang Y,et al.Over-expression of OsDREB genes lead to enhanced drought tolerance in rice.Biotechnol Lett,2008,30(12):2191-2198.
Chen M,Xu Z,Xia L,et al.Cold-induced modulation and functional analyses of the DRE-binding transcription factor gene,GmDREB3,in soybean(Glycine max L.).J Exp Bot,2009,60(1):121-135.
Cheng Z,Targolli J,Huang X Q,et al.Wheat LEA genes,PMA80 and PMA1959,enhance dehydration tolerance of transgenic rice(Oryza sativa L.).Mol Breed,2002,10:71-82.
Cho E K,Hong C B.Over-expression of tobacco NtHSP70-l contributes to drought-stress tolerance in plants.Plant Cel I Rep,2006,25:349-358.
Cominelli E,Galbiati M,Vavasseur A,et al.A guard cell specific MYB transcription factor regulates stomatal movements and plant drought tolerance.Curr Biol,2005,15(13):1196-2000.
Could J,Devery M,Hasegawa 0.Transformation of Zea mays L.using Agrobacterium tumefaciens and the shoot apex.Plant Physiol,1991,95:426-434.
Dai X Y,Xu Y Y,Ma Q B,et al.Overexpression of an R1R2R3 MYB gene,OsMYB3R-2,increases tolerance to freezing,drought,and salt stress in transgenic Arabidopsis.Plant Physiol,2007,143(4):1739-1751.
Delauney A J,Hu C A,Kishor P B,et al.Cloning of ornithine delta-aminotransferase cDNA from Vigna aconitifolia by trans-complementation in Escherichia coli and regulation of proline biosynthesis.J Biol Chem,1993,268(25):18673-18678.
Dubouzet J G,Sakuma Y,Ito Y,Kasuga M,et al.OsDREB genes in rice,Oryza sativa L.,encode transcription activators that function in drought-,high-salt- and cold-responsive gene expression.Plant J,2003,33(4):751-763.
Eulgem T,Rushton PJ,Robatzek S,et al.The WRKY super family of plant transcription factors.Trends Plant Sci,2000,5(5):199-206.
Frame B R,Shou H,Chikwamba R K,et al.Agrobacterium tumefaciens-mediated transformation of maize embryos using a standard binary vector system.Plant Physiol,2002,129(1):13-22.
Fromm M E,Taylor L P,Valbot V.Stable transformation of maize after gene transfer by eletroporation.Nature,1986,319(6056):791-793.
Fujita M,Fujita Y,Maruyama K,et al.A dehydration-induced NAC protein,RD26,is involved in a novel ABA-dependent stress-signaling pathway.The Plant Journal,2004,39:863-876.
Garg A K,Kim J K,Owens T G,et al.Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses.Proc Natl Acad Sci USA,2002,99(25):15898-15903.
Gilmour S J,Sebolt A M,Salazar M P,et al.Over expression of the Arabidopsis CBF3 transcriptional activator mimics multiple biochemical changes associated with cold acclimation.Plant Physiol,2000,124:1854-1865.
Gilmour S J,Zarka D G,Stockinger E J,et al.Low temperature regulation of the Arabidopsis CBF family of AP2 transcriptional activators as an early step in cold-induced COR gene expression.Plant J,1998,16(4):433-442.
Gupta A S,Heinen J L,Holaday A S,et al.Increased resistance to oxidative stress in transgenic plants that overexpress chloroplastic Cu/Zn superoxide dismutase.Proc Natl Acad Sci USA,1993,90(4):1629-1633.
Gutha L R,Reddy A R.Rice DREB1B promoter shows distinct stress-specific responses,and the overexpression of cDNA in tobacco confers improved abiotic and biotic stress tolerance.Plant Mol Biol,2008,68(6):533-555.
Haake V,Cook D,Riechmann J L,et al.Transcription factor CBF4 is a regulator of drought adaptation in Arabidopsis.Plant physiology,2002,130(2):639-648.
Hern(?)ndez G,Ram(?)rez M,Vald(?)s-L(?)pez O,et al.Phosphorus stress in common bean:root transcript and metabolic responses.Plant Physiol,2007,144(2):752-767.
Hiei Y,Ohta S,Komari T,et al.Efficient Transformation of Rice(Oryza Sativa L.)Mediated by Agrobacterium and Sequence Analysis of the Boundaries of the T-DNA.Plant Journal,1994,6(2):271-282.
Hohmann S,Bell W,Neves M J,et al.Evidence for trehalose-6-phosphate-dependent and -independent mechanisms in the control of sugar influx into yeast glycolysis.Mol Microbiol,1996,20(5):981-991.
Holmstrom K O,Welin B,Mandal A,et al.Production of the Escherichia coli betaine-aldehyde dehydrogenase,an enzyme required for the synthesis of the osmoprotectant glycine betaine,in transgenic plants.Plant J,1994,6(5):749-58.
Hsieh T H,Lee J T,Charng Y Y,et al.Tomato plants ectopicaly expressing Arabidopsis CBF1 show enhanced resistance to water deficit stress.Plant Physiology,2002,130(2):618-626.
Ishida Y,Saito H,Ohta S,et al.High efficiency transformation of maize(Zea mays L.)mediated by Agrobacterium tumefaciens.NatBiotechnol,1996,14(6):745-750.
Ishiguro S,Nakamura K.Characterization of a cDNA encoding a novel DNA-binding protein,SPFI,that recognizes SP8 sequences in the 5'upstream regions of genes coding for sporamin and 6-amylase from sweet potato.Mol Gen Genet,1994,244(6):563-571.
Jaglo-Ottosen K R,Gilmour S J,Zarka D G,Schabenberger O,et al.Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance.Science,1998,280(5360):104-106.
Jeanneau M,Gerentes D,Foueillassar X,et al.Improvement of drought tolerance in maize:towards the functional validation of the Zm-Asrl gene and increase of water use efficiency by over-expressing C4-PEPC.Biochimie,2002,84(11):1127-1135.
Jiang C,lu B,Singh J.Requirement of a CCGAC cis-acting element for cold induction of the BN115 gene from,winter Brassica napus.Plant Mol.Biol,1996,30(3):679-684.
Jiang Y,Deyholos M K.Functional characterization of Arabidopsis NaCl-inducible WRKY25 and WRKY33 transcription factors in abiotic stresses.Plant Mol Biol,2009,69(1-2):91-105.
Jin H,Martin C.Multifunctionality and diversity within the plant MYB-gene family.Plant Mol Biol,1999,41(5):577-585.
Kang J Y,Choi H I,Im M Y,et al.Arabidopsis basic leucine zipper proteins that mediate stress-responsive abscisic acid signaling.Plant Cell,2002,14(2):343-357.
Katiyar-Agarwal S,Agarwal M,Grover A.Heat-tolerant basmati rice engineered by over-expression of hsp101.Plant Mol Biol,2003,51(5):677-686.
Kenward K D,Altschuler M,Hildebrand D,et al.Accumulation of type I fish antifreeze protein in transgenic tobacco is cold-spesific.Plant Mol Biol,1993,23(2):377-385.
Kishor PBK,Hong Z,Miao G H,et al.Over expression of[delta]-Pyrroline-5 -Carboxylate Synthetase Increases Proline Production and Confers Osmotolerance in Transgenic Plants.Plant Physiol,1995,108(4):1387-1394.
Klein T M,Kornstein L,Sanford J C,et al.Transformation of Maize Cells by Particle Bombardment.Plant Physiology,1989,91(1):440-444.
Koziel M G,Beland G L,Bowman C,et al.Field performence of elite transgenic maize plant expressing all insecticidal protein derived from Bacillus thuringiensis.Bio/Technolgy,1993,11:194-200.
Lea U S,Slimestad R,Smedvig P,et al.Nitrogen deficiency enhances expression of specific MYB and bHLH transcription factors and accumulation of end products in the flavonoid pathway.Planta,2007,225(5):1245-1253.
Lee S C,Choi H W,Hwang I S,et al.Functional roles of the pepper pathogen-induced bZIP transcription factor,CAbZIPl,in enhanced resistance to pathogen infection and environmental stresses.Planta,2006,224(5):1209-1225.
Li S,Fu Q,Huang W,Yu D.Functional analysis of an Arabidopsis transcription factor WRKY25 in heat stress.Plant Cell Rep,2009,Epub ahead of print.Lipsick J S.One billion years of Myb.Oncogene,1996,13(2):223-235.
Liu Q,Kasuga M,Sakuma Y,et al.Two transcription factors,DREBl 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,1998,10:1391-1406.
Liu Y,Wang G,Liu J,et al.Transfer of E.coli gutD gene into maize and regeneration of salt-tolerant transgenic plants.Sci China C Life Sci,1999,42(1):90-95.
Lu G,Gao C,Zheng X,et al.Identification of OsbZIP72 as a positive regulator of ABA response and drought tolerance in rice.Planta,2009,229(3):605-615.
Lung J,Giraudat J.Abscisic acid signal transduction.Annu Rev.Plant Physiol Plant Mol Biol,1998,49:199-222.
Lupotto E,Reali A,Oassera S,et al.Maize elite inbred lines are susceptible to Agrobacterium tumefaciens-mediated transformation.Maydica,1999,44(3):211-218.
Martin C,Paz-Ares J.MYB transcription factors in plants.Trends Genet,1997,13(2):67-73.
Medina J,Bargues M,Terol J,et al.The Arabidopsis CBF gene family composed of three gene encoding AP2 domain-containing proteins whose expression is regulated by low temperature but not by abscisic acid or dehydration.Plant Physiol,1999,119:463-469.
Miyake K,Ito T,Sends M.Isolation of a subfamily of genes for R2R3-MYB transcription factors showing up-regulated expression under nitrogen nutrient-limited conditions,Plant Mol Biol,2003,53(1-2):237-245.
Moffat A S.Finding New Ways to Protect Drought-Stricken Plants.Science,2002,296(5571):1226-1229.
Moghaieb REA,Tanaka N.Sancoka H,et al.Expression of betaine aldehyde dehydrogenase gene in transgenic tomato hairy roots leads to the accumulation of glycine betaine and contributes to the maintenance of the osmotic potential under salt stress.Soil Sci Plant Nutr,2000,46(4):873-883.
Nanjo T,Kobayashi M,Yoshiba Y,et al.Antisense suppression of proline degradation improves tolerance to freezing and salinity in Arabidopsis thaliana.FEBS Lett,1999,461(3):205-210.
Ohnishi T,Sugahara S,Yamada T,et al.OsNAC6,a member of the NAC gene family,is induced by various stresses in rice.Genes Genet Syst,2005,80:135-139.
Ouellet F,Vazquez-Tello A,Sarhan F.The wheat Wcs120 promoter is cold-inducible in both monocotyledonous and dicotyledonous species.FEBS Letters,1998,423(3):324-328.
Pardo J M,Reddy M P,Yang S,et al.Stress signaling through Ca2+/calmodulin-dependent protein phosphatase calcineurin mediates salt adaptation in plants.Proc Natl Acad Sci USA,1998,95(16):9681-9686.
Park E J,Jeknic Z,Sakamoto A,et al.Genetic engineering of glycinebetaine synthesis in tomato protects seeds,plants,and flowers from chilling damage.Plant J,2004,40(4):474-487.
Perl A,Perl-Treves R,Galili S,et al.Enhanced oxidative stress defense in transgenic potato expressing tomato Cu/Zn superoxide dismutases.Theor Appl Genet,1993,85(5):568-576.
Qin F,Kakimoto M,Sakuma Y,et al.Regulation and functional analysis of ZmDREB2A in response to drought and heat stresses in Zea mays L.Plant J,2007,50(1):54-69.
Qin F,Sakuma Y,Li J,et al.Cloning and functional analysis of a novel DREB1/CBF transcription factor involved in cold-responsive gene expression in Zea mays L.Plant Cell Physiol,2004,45(8):1042-1052.
Quan R,Shang M,Zhang H,et al.Engineering of enhanced glycine betaine synthesis improves drought tolerance in maize.Plant Biotechnol J,2004,2(6):477-486.
Rabinowicz P D,Braun E L,Wolfe A D,et al.Maize R2R3 MYB gene:sequence analysis reveals amplification in the higher plants.Genetics,1999,153(1):427-444.
Rhodes C A,Pierce D A,Mettler I J,et al.Genetically transformed maize plants from protoplast.Sciene,1988,240(4849):204-207.
Riechmann J L,Heard J,Martin G,et al.Arabidopsis transcription factors:genome-wide comparative analysis among Eukaryotes.Science,2000,290(5499):2105-2110.
Riechmann J L,Meyerowitz E M.The AP2/EREBP family of plant transcription factors.Biol Chem,1998,379(6):633-646.
Rubio M C,Gonzalez E M,Minchin F R,et al.Effects of water stress on antioxidant enzymes of leaves and nodules of transgenic alfalfa overexpressing superoxide dismutases.Physiol Plant,2002,115(4):531-540.
Saijo Y,Hata S,Kyozuka J,et al.Over-expression of a single Ca~(2+)-dependent protein kinase confers both cold and salt/drought tolerance on rice plants.Plant J,2000,23(3):319-327.
Sakuma Y,Liu Q,Dubouzet J G,et al.DNA-binding specificity of the ERF domain of Arabidopsis DREBs.transcription factors involved in dehydration and cold-inducible gene expression.Biochem Biophys Res Commun,2002,290(3):998-1009.
Sanmiya K,Suzuki K,Egawa Y,Shono M.Mitochondrial small heat-shock protein enhances thermotolerance in tobacco plants.FEBS Lett,2004,557(1-3):265-268.
Sayari AH,Bouzid R G,Bidani A,et al.Overexpression of Al-pyrroline-5-carboxylate synthetase increases proline production and confers salt tolerance in transgenic potato plants.2005,169(4):746-752.
Selth L A,Dogra S C,Rasheed M S,et al.A NAC domain protein interacts with tomato leaf curl virus replication accessory protein and enhances viral replication.Plant Cell,2005,17(1):311-325.
Shen B,Jensen R G,Bohnert H J.Increased resistance to oxidative stress in transgenic plants by targeting mannitol biosynthesis to chloroplasts.Plant Physiol.1997,113(4):1177-83.
Shinwari Z K,Nakashima K,Miura S,et al.An Arabidopsis gene family encoding DRE/CRT binding proteins involved in low-temperature-responsive gene expression.Biochem Biophys Res Commun,1998,250(1):161-170.
Shou H,Bordallo P and Wang K.Expression of the Nicotiana protein kinase(NPK1)enhanced drought tolerance in transgenic maize.J Exp Bot,2004,55(399):1013-1019.
Sivamani E,Bahieldin A,Wraith J M,et al.Improved biomass productivity and water use efficiency under water deficit conditions in transgenic wheat constitutively expressing the barley HVA1 gene.Plant Sci,2000,155(1):1-9.
Slooten L,Capiau K,Camp W V,et al.Factors affecting the enhancement of oxidative stress tolerance in transgenic tobacco overexpressing manganese superoxide dismutase in the chloroplasts.Plant Physiol,1995,107(3):737-775.
Stockinger E J,Gilmour S J,Thomashow M F.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,1997,94(3):1035-1040.
Stockinger E J,Mao Y,Regier M K,et al.Transcriptional adaptor And histon acetyltransferase proteins in Arabidopsis and their interactions with CBF1,a transcriptional adaptor involved in cold-regulated gene expression.Nucleic Acids Res,2001,29(7):1524-1533.
Stracke R,Werber M,Weisshaar B.The R2R3-MYB gene family in Arabidopsis thaliana.Curr Opin Plant Biol,2001,4(5):447-456.
Sun W,Van Montagu M,Verbruggen N.Small heat shock proteins and stress tolerance in plants.Biochim Biophys Acta,2002,1577(1):1-9.
Tang W,Peng X,Newton R J.Enhanced tolerance to salt stress in transgenic loblolly pine simultaneously expressing two genes encoding mannitol-1-phosphate dehydrogenase and glucitol-6-phosphate dehydrogenase.Plant Physiol Biochem,2005,43(2):139-46.
Tran L S,Nakashima K,Sakuma Y,et al.Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 promoter.Plant Cell,2004,6(9):2481-2498.
Tran LS,Nakashima K,Sakuma Y,et al.Isolation and functional analysis of Ambidopsis stress-inducible NAC transcription factors that bind to a drought responsive Cis-element in the EARLY RESPONSIVE TO DEHYDRATION STRESS promoter.Plant Cell,2004,16(9):2481-2498.
Tyerman S D,Niemietz C M,Bramley H.Plant aquaporins:multifunctional water and solute channels with expanding roles.Plant Cell Environ,2002,25(2):173-194.
Ulker B,Somssich I E.WRKY transcription factors:from DNA binding towards biological function.Curr Opin Plant Biol,2004,7(5):491-498.
Umezawa T,Fujita M,Fujita Y,et al.Engineering drought tolerance in plants:discovering and tailoring genes to unlock the future.Curr Opin Biotechnol,2006,17(2):113-122.
Umezawa T,Yoshida R,Maruyama K,et al.SRK2C,a SNFl-related protein kinase 2,improves drought tolerance by controlling stress-responsive gene expression in Arabidopsis thaliana.Proc Natl Acad Sci USA,2004,101(49):17306-17311.
Vannini C,Locatelli F,Bracale M,et al.Overexpression of the rice Osmyb4 gene increases chilling and freezing tolerance of Arabidopsis thaliana plants.Plant J,2004,37(11):115-127.
Vendniscolo E C,Schuster I,Pileggi M,et al.Stress-induced synthesis of proline confers tolerance to water deficit in transgenic wheat.J Plant Physiol,2007,164(10):1367-1376.
Verling E.The roles of heat shock proteins in plants.Annu Rev Plant Physiol Mol Biol,1991,42:579-620.Wang C R,Yang A F,Yue G D,et al.Enhanced expression of phospholipase C 1(ZmPLCl)improves drought tolerance in transgenic maize.Planta,2008,227(5):1127-1140.
Wang J W,Yang F P,Chen X Q,et al.Induced expression of DREB transcriptional factor and study on its physiological effects of drought tolerance in transgenic wheat.Acta Genet Sin,2006,33:468-476.
Wang L,Li X,Chen S,Liu G Enhanced drought tolerance in transgenic Leymus chinensis plants with constitutively expressed wheat TaLEA3.Biotechnol Lett,2009,31(2):313-319.
Wang L,Zhao C M,Wang Y J,et al.Overexpression of chloroplast-localized small molecular heat-shock protein enhances chilling tolerance in tomato plant.J Plant Physiol Mol Biol,2005,31(2):167-174.
Wang W,Vinocur B,Shoseyov O,et al.Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response.Trends Plant Sci,2004,9(5):244-52.
Wu X,Shiroto Y,Kishitani S,et al.Enhanced heat and drought tolerance in transgenic rice seedlings overexpressing OsWRKY11 under the control of HSP101 promoter.Plant Cell Rep,2009,28(1):21-30.X
iong L,Schumaker K S,Zhu J K.Cell signaling during cold,drought,and salt stress.Plant Cell,2002,14Suppl:S165-183.
Xu Z S,Ni Z Y,Li Z Y,et al.Isolation and functional characterization of HvDREBI-a gene encoding a dehydration-responsive element binding protein in Hordeum vulgare.J Plant Res,2009,122(1):121-130.
Yamaguchi-Shinozaki K,Shinozaki K.Improving plant drought,salt and freezing tolerance by gene transfer of a single stress-inducible transcription factor.Novartis Found Symp,2001,236:176-186.
Yamaguchi-Shinozaki K,Shinozaki K.Transcriptional regulatory networks in cellular responses and toleranse to dehydration and cold stress.Annu.Rev.Plant Biol,2006,57:781-803.
Yancey P H.Living with water stress:evolution of osmolyto system.Science,1982,217:1214-1222.
Yokotani N,Ichikawa T,Kondou Y,et al.Tolerance to various environmental stresses conferred by the salt-responsive rice gene ONAC063 in transgenic Arabidopsis.Planta,2009,ahead of print.
Zhao Z Y,Gu W N,Cai T,et al.High throughput genetic transformation mediated by Agrobacterium tumefaciens in maize.Mol.Breed,2002,8(4):323-333.
Zhu J K.Salt and drought stress signal transduction in plants.Annu Rev Plant Biol,2002,53:247-73.
丁群星,谢友菊,戴景瑞等,用子房注射法将Bt毒蛋白基因导入玉米的研究.中国科学(B辑),1993,23(7):707-713.
傅桂荣,张初,赵晓祥等.转美州拟鲽抗冻蛋白基因(AFP)番茄的叶片电导率测定.生物技术,1997,7(5):7-8.
高银.植物抗逆机制与基因工程研究进展.内蒙古农业科技,2007,5:75-78.
黄璐,卫志明.农杆菌介导的玉米遗传转化.实验生物学报,1999,32(4):381-389.
李芳,李先文,黎定军.AP2/EREBP转录因子及其在植物抗逆中的作用.江西农业学报,2008,20(1):44-47.
李慧芬,刘翔,曲强等.转抗虫融合基因(cry1Ac3-cpti)玉米(Zea mays L).自然科学进展,2002,12(1):37-41.
李余良,胡建广,苏菁,刘建华.子房注射法将BI基因导入超甜玉米.玉米科学,2005,13(1):41-43.
李跃强,宣维健,盛承发.植物的低温蛋白.生态学报,2004,249(5):1034-1039.
梁峥,马德饮,汤岚等.菠菜甜菜碱醛脱氢酶基因在烟草中的表达.生物工程学报,1997,(13):236-240.
粱慧敏,夏阳,王太明.植物抗寒冻、抗旱、耐盐基因工程研究进展.草业学报,2003,12(3):1-7.
林茂,闫海霞,眭顺照,李名扬.植物CBF转录因子及其在基因工程中的作用.广西农业科学,2008,39(1):21-25.
林士杰,李俊涛,姜静等.转柽柳晚期胚胎富集蛋白基因烟草的耐低温性分析.生物技术通讯,2006,14(4):563-566.
刘蕾,杜海,唐晓风等.MYB转录因子在植物抗逆胁迫中的作用及其分子机理.遗传,2008,30(10):1265-1271.
刘万勃,宋明,张中灵等.抗冻蛋白的研究进展.生物学杂志,2000,17(4):7-8.
刘欣,李云.转录因子与植物抗逆性研究进展.中国农学通报,2006,22(4):61-66.
刘昀,李冉辉,郑易之等.大豆PM2蛋白及其结构域可提高烟草耐盐性.深圳大学学报:理工版,2007,24(1):95-101.
卢东长城,利用共抑制技术获得高直链淀粉含量的玉米新种质.2008,首都师范大学硕士论文.
路子显,常团结,刘翔等.植物碱性亮氨酸拉链(bZIP)蛋白的研究进展(一)--结构、分类、分布和同源性分析.遗传,2001,23:564-570.
孟凤,郁松林,郊强卿等.甜菜碱与植物抗逆性关系之研究进展.中国农学通报,2008,24(4):225-228.
彭运生,刘恩.关于提取叶绿素方法的比较研究.北京农业大学学报,1992,18(3):247-250.
祁永红.大豆DNA直接导入玉米自交系的研究.玉米科学,2000,8(1):34-36.
钱刚,翟旭光,韩兆雪等.西藏青稞IEA3蛋白新抗旱基因的克隆与序列分析.作物学报,2007'33(2):292-296.
权瑞党,尚梅,王兆玉,张举仁.农杆菌介导的雪花莲凝集素基因转入玉米骨干自交系.西北植物学报,2004,24(5):761-767.
荣红颖,外源脱水应答转录因子DREB1B在转基因小麦后代的稳定表达研究.2008,首都师范大学硕士论文.
苏琦,尚宇航,杜密英,杜进民.植物WRKY转录因子研究进展.农业生物技术科学,2007,23(5):94-98.
孙红伟,朱常香,杨崇良等.玉米基因枪转化Bt基因的研究.山东农业科学,2004,3:20-22.
汤章城.逆境条件下植物脯氨酸累积及其可能的意义.植物生理学通讯,1984(1):15-21.
王罡,杜娟,张艳华.用花粉管通道法将Bt杀虫基囚导入玉米自交系的研究.玉米科学,2002,10(1):36-37.
王国莉,郭振飞.植物耐冷性分子机理的研究进展.植物学通报,2003,20(6):671-679.
王国英,杜天兵,张宏等.用基因枪将Bt毒蛋白基因转入玉米及转基因植株再生.中国科学(B辑),1995,25(1):1-7.
王书子,吴少辉,高海涛等.旱地小麦品种筛选鉴定及其形态特征探讨干旱地区农业研究.,2001,19(2):76-80
武明宇,郝楠.玉米抗旱性的研究进展.现代农业科技,2008,9:124-127.
武模戈,袁刘正.多胺与植物抗逆性关系研究进展.安徽农业科学,2008,36(9):3516-3518.
杨淑慎,山仑,郭蔼光等.水通道蛋白与植物抗旱性.干旱地区农业研究,2005,23(6):214-218.
张明生,谢波,谈锋.甘薯可溶性蛋白、叶绿素及ATP含量变化与品种抗旱性关系的研究.中国农业科学,2003,36(1):13-16.
张淑娟.转盐芥CBF1基因提高玉米抗逆性的研究.2007,山东大学硕士论文.
张艳贞,季静,张领兵等.以基因枪法将BT杀虫蛋白基因导入常规玉米自交系的研究.玉米科学,2001,9(4):23-26.
张正斌.作物抗旱节水的生理遗传育种基础.北京:科学出版社,2003.
周宜君,冯金朝,马文文等.吕惠娟.植物抗逆分子机制研究进展.中央民族大学学报(自然科学版),2006,15(2):169-176.
邹杰,陈信波,刘爱玲等.植物热激蛋白与作物非生物抗逆性的改良.植物生理学通讯,2007,43(5):981-985.