农杆菌介导水稻RdreB1BI基因转化草莓的研究
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摘要
草莓(Fragaria ananassa Duch)属于蔷薇科草莓属多年生草本植物,是一种重要的经济浆果,在世界范围内广泛种植。然而草莓安全越冬和耐寒品种的缺乏成为生产上的突出问题。生物技术的飞速发展为解决这一问题提供了新的途径。水稻转录因子RdreB1BI是一个调控植物发育且与植物胁迫抗性相关的转录因子。已有研究证明将RdreB1BI基因转化拟南芥,拟南芥植株的抗早性和耐寒性有很大提高。到目前为止还未见有RdreB1BI基因转化草莓的研究。因此本试验目的是用根癌农杆菌介导的转化方法将水稻RdreB1BI基因导入草莓细胞,获得抗寒的转基因植株。主要研究结果有以下几点:
1.以草莓‘丰香'、‘雪蜜'、‘鬼怒甘'的叶片和叶柄为外植体,建立了高效离体再生体系。‘丰香'叶片和叶柄的最佳不定芽再生培养基为:MS+TDZ(2.0mg/L)+IBA(0.2mg/L)+2,4-D(0.1mg/L)和MS+B5+TDZ(2.0mg/L)+IBA(0.2mg/L)+2,4-D(0.1mg/L)。‘雪蜜'叶片和叶柄的最佳不定芽再生培养基为:MS+TDZ(1.5mg/L)+IBA(0.2mg/L)和MS+B5+TDZ(2.0mg/L)+IBA(0.2mg/L)。‘鬼怒甘'叶片和叶柄的最佳不定芽再生培养基为:MS+BA(2.0mg/L)+IBA(0.1mg/L)和MS+BA(2.0mg/L)+IBA(0.1mg/L);‘丰香'、‘雪蜜'暗培养时间为14天,‘鬼怒甘'暗培养时间为7天;‘丰香'、‘鬼怒甘'叶片不定芽再生的最佳叶龄为10-30天,‘雪蜜'叶片不定芽再生的最佳叶龄为20天以下。三个草莓品种叶柄不定芽再生的最佳叶龄都是20天以下;叶片放置方式对三个草莓品种的叶片不定芽再生没有影响,叶柄刻伤向上放置有利于三个草莓品种叶柄不定芽再生。
2.克隆了rd29A启动子,通过对比分析,与已公布的序列有98.72%的同源性,主要调控区域也一致。构建了两个植物表达载体pYH4215-rd29A和pYF7713-Rdre。
3.通过研究影响遗传转化的因素,建立了农杆菌介导的‘雪蜜'草莓遗传转化体系:菌液浓度为OD600=0.4;侵染时间为9min;选用200mg/L的Cb作为抑菌抗生素,20mg/L的Kan作为筛选抗生素;共培养时间3天;共培养培养基:MS+TDZ(2.0mg/L)+IBA(0.2 mg/L);筛选培养基:MS+TDZ(2.0mg/L)+IBA(0.2mg/L)+Kan(20mg/L)+Cb(200mg/L);伸长培养基:MS+6-BA(0.5mg/L)+NAA(0.1mg/L)+Kan(20mg/L)+Cb(200mg/L);生根培养基:MS+IBA(0.1mg/L)+Kan(20mg/L)+Cb(200mg/L)。
4.获得了Kan抗性的含有RdreB1BI基因的草莓转基因株系15株,PCR以及RT-PCR检测证明,有4个株系外源基因已经成功的导入和整合到草莓基因组中。脯氨酸含量的测定和低温处理表明,转基因的植株较非转基因植株抗寒性有所提高。
Strawberry(Fragaria ananassa Duch),perennial herbaceous plant in the Rosaceae family,which is a major berry crop around the world.However,safe hibernation and deficiency of cold tolerance varieties are outstanding problems.In this case,biotechnology which developed speedly is an alternative efficient strategy to solve the problems. RdreB1BI gene is important transcription factors which related to plant development and physiology.It was investigated that RdreB1BI transcription factors are able to improve tolerance to drought,high-salt and cold stresses in Arabidopsis.But there is no research in transformation of strawberry with RdreB1BI gene of rice at present.So in this study, RdreB1BI was used as purpose gene,the transformation of strawberry by Agrobacterium-mediated was studied.The experiment research results were as follows:
1.The ideal adventitious shoot regeneration system of strawberry 'Toyonoka','Xuemi' and 'Guinugan' by leaf and petiole explants were established In this study.The results showed that the highest adventitious shoot regeneration rate of Toyonoka's leaf disc was obtained on MS medium supplemented with 2.0mg/L TDZ,0.2 mg/L IBA and 0.1mg/L 2,4-D.The highest adventitious shoot regeneration rate of Toyonoka's petiole was obtained on MS+B5 medium supplemented with 2.0mg/L TDZ,0.2mg/L IBA and 0.1mg/L 2,4-D. The highest adventitious shoot regeneration rate of Xuemi's leaf disc was obtained on MS medium supplemented with 1.5mg/L TDZ and 0.2mg/L IBA.The highest adventitious shoot regeneration rate of Xuemi's petiole was obtained on MS+B5 medium supplemented with 2.0 mg/L TDZ,0.2mg/L IBA.The highest adventitious shoot regeneration rate of Guinugan's leaf disc was obtained on MS medium supplemented with 2.0mg/L BA and 0.1mg/L IBA.The highest adventitious shoot regeneration rate of Guinugan's petiole was obtained on MS medium supplemented with 2.0 mg/L BA and 0.2mg/L IBA.For 'Toyonoka' and 'Xuemi' Dark treatment for two weeks could increase the adventitious shoot regeneration.But for 'Guinugan' Dark treatment for one week could increase the adventitious shoot regeneration.The ideal leaf ages was 10 to 30 days of 'Toyonoka' and 'Guinugan' leaf.The ideal leaf ages was 10 to 20 days of 'Xuemi' leaf.The ideal leaf ages was 10 to 20 days of the three varieties petiole.There was no effect of leaf burl on adventitious shoot regeneration of strawberry.
2.The region of rd29A gene promoter was amplified by PCR technique.Sequence analysis showed that the fragment contained the main regulation motifs.Two a-ntisense expression vectors called pYH4215-rd29A and pYF7713-Rdre were obtained.
3.By optimizing the transformation system of strawberry,a high-efficient transformation system was established.The bacterial concentration of OD600=0.4 and 10 minutes was the best condition.The available concentration of Cb was 200mg/L to eliminate Agrobacterium tumefaciens.The corresponding concentration of Kan was 20mg/L to screen.Co-culture for 3 days.The optimum Co-culture medium was MS+TDZ(2.0mg/L)+IBA(0.2mg/L),the optimum medium for elongation was MS+6-BA(0.5mg/L)+NAA(0.1mg/L)+Kan(20mg/L)+Cb(200mg/L),the optimum root inducing medium was MS+IBA(0.1mg/L)+Kan(20mg/L)+Cb(200mg/L).
4.The transgenic plants contained RdreB1BI were obtained.The result of PCR and RT-PCR showed that the target gene was successfully integrated into the genome of strawberry.The concentration of praline indicated cold resistance of transformation strawberry had improved.
1.以草莓‘丰香'、‘雪蜜'、‘鬼怒甘'的叶片和叶柄为外植体,建立了高效离体再生体系。‘丰香'叶片和叶柄的最佳不定芽再生培养基为:MS+TDZ(2.0mg/L)+IBA(0.2mg/L)+2,4-D(0.1mg/L)和MS+B5+TDZ(2.0mg/L)+IBA(0.2mg/L)+2,4-D(0.1mg/L)。‘雪蜜'叶片和叶柄的最佳不定芽再生培养基为:MS+TDZ(1.5mg/L)+IBA(0.2mg/L)和MS+B5+TDZ(2.0mg/L)+IBA(0.2mg/L)。‘鬼怒甘'叶片和叶柄的最佳不定芽再生培养基为:MS+BA(2.0mg/L)+IBA(0.1mg/L)和MS+BA(2.0mg/L)+IBA(0.1mg/L);‘丰香'、‘雪蜜'暗培养时间为14天,‘鬼怒甘'暗培养时间为7天;‘丰香'、‘鬼怒甘'叶片不定芽再生的最佳叶龄为10-30天,‘雪蜜'叶片不定芽再生的最佳叶龄为20天以下。三个草莓品种叶柄不定芽再生的最佳叶龄都是20天以下;叶片放置方式对三个草莓品种的叶片不定芽再生没有影响,叶柄刻伤向上放置有利于三个草莓品种叶柄不定芽再生。
2.克隆了rd29A启动子,通过对比分析,与已公布的序列有98.72%的同源性,主要调控区域也一致。构建了两个植物表达载体pYH4215-rd29A和pYF7713-Rdre。
3.通过研究影响遗传转化的因素,建立了农杆菌介导的‘雪蜜'草莓遗传转化体系:菌液浓度为OD600=0.4;侵染时间为9min;选用200mg/L的Cb作为抑菌抗生素,20mg/L的Kan作为筛选抗生素;共培养时间3天;共培养培养基:MS+TDZ(2.0mg/L)+IBA(0.2 mg/L);筛选培养基:MS+TDZ(2.0mg/L)+IBA(0.2mg/L)+Kan(20mg/L)+Cb(200mg/L);伸长培养基:MS+6-BA(0.5mg/L)+NAA(0.1mg/L)+Kan(20mg/L)+Cb(200mg/L);生根培养基:MS+IBA(0.1mg/L)+Kan(20mg/L)+Cb(200mg/L)。
4.获得了Kan抗性的含有RdreB1BI基因的草莓转基因株系15株,PCR以及RT-PCR检测证明,有4个株系外源基因已经成功的导入和整合到草莓基因组中。脯氨酸含量的测定和低温处理表明,转基因的植株较非转基因植株抗寒性有所提高。
Strawberry(Fragaria ananassa Duch),perennial herbaceous plant in the Rosaceae family,which is a major berry crop around the world.However,safe hibernation and deficiency of cold tolerance varieties are outstanding problems.In this case,biotechnology which developed speedly is an alternative efficient strategy to solve the problems. RdreB1BI gene is important transcription factors which related to plant development and physiology.It was investigated that RdreB1BI transcription factors are able to improve tolerance to drought,high-salt and cold stresses in Arabidopsis.But there is no research in transformation of strawberry with RdreB1BI gene of rice at present.So in this study, RdreB1BI was used as purpose gene,the transformation of strawberry by Agrobacterium-mediated was studied.The experiment research results were as follows:
1.The ideal adventitious shoot regeneration system of strawberry 'Toyonoka','Xuemi' and 'Guinugan' by leaf and petiole explants were established In this study.The results showed that the highest adventitious shoot regeneration rate of Toyonoka's leaf disc was obtained on MS medium supplemented with 2.0mg/L TDZ,0.2 mg/L IBA and 0.1mg/L 2,4-D.The highest adventitious shoot regeneration rate of Toyonoka's petiole was obtained on MS+B5 medium supplemented with 2.0mg/L TDZ,0.2mg/L IBA and 0.1mg/L 2,4-D. The highest adventitious shoot regeneration rate of Xuemi's leaf disc was obtained on MS medium supplemented with 1.5mg/L TDZ and 0.2mg/L IBA.The highest adventitious shoot regeneration rate of Xuemi's petiole was obtained on MS+B5 medium supplemented with 2.0 mg/L TDZ,0.2mg/L IBA.The highest adventitious shoot regeneration rate of Guinugan's leaf disc was obtained on MS medium supplemented with 2.0mg/L BA and 0.1mg/L IBA.The highest adventitious shoot regeneration rate of Guinugan's petiole was obtained on MS medium supplemented with 2.0 mg/L BA and 0.2mg/L IBA.For 'Toyonoka' and 'Xuemi' Dark treatment for two weeks could increase the adventitious shoot regeneration.But for 'Guinugan' Dark treatment for one week could increase the adventitious shoot regeneration.The ideal leaf ages was 10 to 30 days of 'Toyonoka' and 'Guinugan' leaf.The ideal leaf ages was 10 to 20 days of 'Xuemi' leaf.The ideal leaf ages was 10 to 20 days of the three varieties petiole.There was no effect of leaf burl on adventitious shoot regeneration of strawberry.
2.The region of rd29A gene promoter was amplified by PCR technique.Sequence analysis showed that the fragment contained the main regulation motifs.Two a-ntisense expression vectors called pYH4215-rd29A and pYF7713-Rdre were obtained.
3.By optimizing the transformation system of strawberry,a high-efficient transformation system was established.The bacterial concentration of OD600=0.4 and 10 minutes was the best condition.The available concentration of Cb was 200mg/L to eliminate Agrobacterium tumefaciens.The corresponding concentration of Kan was 20mg/L to screen.Co-culture for 3 days.The optimum Co-culture medium was MS+TDZ(2.0mg/L)+IBA(0.2mg/L),the optimum medium for elongation was MS+6-BA(0.5mg/L)+NAA(0.1mg/L)+Kan(20mg/L)+Cb(200mg/L),the optimum root inducing medium was MS+IBA(0.1mg/L)+Kan(20mg/L)+Cb(200mg/L).
4.The transgenic plants contained RdreB1BI were obtained.The result of PCR and RT-PCR showed that the target gene was successfully integrated into the genome of strawberry.The concentration of praline indicated cold resistance of transformation strawberry had improved.
引文
[1]刘祖棋,张石城.植物抗性生理学[M].北京:中国农业出版社,1994,43-86
[2]王小菁,李娘辉,潘瑞炽.植物生理学[M].北京:高等教育出版社,2006,432-476
[3]Weiser C J.Cold resistance and injury in woody plants[J].Science,1979,169:1269-1273
[4]Briggs D R.The chemistry of the living bark of the black locust tree in relation to frost hardiness[J].Arch.Biochem.,1949,23:8-11
[5]Hughes M A,Dunn M A.The molecular biology of plant acclimation to low temperature[J]Experimental Botany,1996,47(296):291-296
[6]Danyluk J,Houde M,Rassart E,et al.Differential expression of a gene encoding an acidic dehydrin in chilling sensitive and freezing-tolerant gramineae species[J].FEBS Letters,344,20-22
[7]Murata N,Yamaya J.Molecular species composition of phosphati-dylgylcerols from chilling semsitive and chilling resistance plants[J].Plant Cell Physiol,1983,24:81-84
[8]Guy C L,Haskell D,Neven L,et al.Llydratioirstate-resposive proteins link cold and droughtstress in spinach[J].Planta,1992,188:265-268
[9]Liu Qing,Kasuga M,Sakuma Y,et al.Two transcription factors,DREB1 and DREB2,with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought and low-temperatuxe-responsive gene expression respectively,in Arabidopsis[J].Plant Cell,1998,10(8):1391-1396
[10]Yamaguchi-Shinozaki K,Shinozaki K.A novel cis-acting element in an Arabidopsis Gene is involved in responsiveness to drought,low-temperature,or high-salt stress[J].Plant Cell,1994,6(2):251-264
[11]马建忠.植物的冷诱导基因[J].农业生物技术学报,1996,4(1):8-10
[12]余叔文,汤章城.植物生理与分子生物学[M].北京:科学出版社,1999,235-278
[13]Gibson S,Arondel V.Cloning of a temperature-regulated gene encoding a chloroplast-3 desaturase from Arabidopsis thaliana[J].Plant Physiol,1994,106:1615-1619
[14]Kodama H,Horiguchi G.Fatty acid desaturation during chilling acclimation is one of the factors involved in conferring low temperature tolerance to young tobacco leaves[J].Plant Physiology,1995,107(4):1177-1179
[15]Wada H,Gombos Z,Murata N.Enhancement of chilling tolerance of a cyanobacterium by genetic manipulaton of fatty acid desaturation[J].Nature,1990,347(13):200-205
[16]王艇,苏应娟,刘良式,等.植物低温诱导蛋白和低温诱导基因的表达调控[J].武汉植物学研究,1997,15(1):80-90
[17]Jarillo J A,Capei J,Leyva A,et al.Two related low-temperature-induced genes of Arabidopsis encode proteins showing high homology to 14-3-3 proteins,a family of putative Kinase regulators[J].Plant Molecular Biology,1994,25:693-696
[18]Lin C,Thomashow M F.A cold-regulated Arabidopsis gene encodes a polypetide having potent cryoprotective activity[J].Biochem.Biophysic.Res.Com..,1992,183:1103-1107
[19]江勇,贾士荣,费云标,等.抗冻蛋白及其在植物抗冻生理中的作用[J].植物学报,1999,7(41):678-182
[20]Antikainen M,Griffith M.Antifreeze protein accumulation in freezing tolerant cereals[J].Phy s-iologia Plantarum,1997,99:423-426
[21]Kasuga M,Liu Qi,Mura S.et al.Improving plant drought,salt,and freezing tolerance by gene transfer of a single stress- inducible transcription factor[J].Nature Biotechnologi,1999,17:287-293
[22]Kenward K D,Altschuler M,Hildebrand D.et al.Accumulation of type Ⅰ fish antifreeze protein in transgenic tobacco is cold-specific[J].Plant Molecular Biology,1993,23:377-382
[23]Guy L C.Altered gene expression during cold acclimation of spinach[J].Proc.Natl.Acad.Sci.USA,1985,82:3673-3677
[24]Autus N N.Constitutive expression of the cold-regulated Arabidopsis thsaliana cor15a gene affects both chloroplast feezing to tolerance[J].Proc.Natl.Acad.Sci.USA,1996,93:13404-1409
[25]Jaglo-Ottoseu K R.Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance[J].Sience,1998,280(5360):104-106
[26]Wallis J.Expression of synthetic antifreeze protein in potato reduces electrolyte release at freezing temperatures[J].Plant Molecular Biology,1997,35(3):323-330
[27]Culter A J.Winter flounder antifreeze protein improves the cold hardiness of plant tissue[J].Plant Physiol,1989,135(3):351-354.
[28]万巧兰.柑橘离体形态建成及转鱼类抗冻蛋白基因研究[D].北京:中国农业科学院硕士学位论文,1996
[29]Kenward K D.Type Ⅱ fish antifreeze protein accumulation in transgenic tobacco does not confer frost resistance[J].Transgenic Research,1999,8(2):105-117
[30]Georges F.Design and cloning of a synthetic gene for the flounder antifreeze protein and its expression in plant cell[J].Gene,1990,91:159-165
[31]黄永芬,汪清胤,付桂荣,等.美洲拟蝶抗冻蛋白基因(afps)导入西红柿的研究[J].生物化学杂志,1997,13(4):418-422
[32]Chakrabartty.Structure function relationships in a winter flounder antifreeze polypeptide[J].J.Biol.Chem.,1989,264(19):11307-11312
[33]Lee JS.The redaction of the freezing point of tobacco plants transformed with the gene encoding for the antifreeze protein from winter flounder[J].J.Cell.Biochem.,1990,14(supple):E303
[34]Griffith M.Antifreeze protein produced endogenously in winter rye leaves[J].Plant Physiol 1992,100:593-596
[35]Mckersis B D,Chen Y.Superoxide dismutase enbances tolerance of freezing stress in transgenic alfalfa(Medicago sative L.)[J].Plant Physiol,1993,103(4):1155
[36]赵微平.植物基因组构建、表达和调控[M].北京:首都师范大学出版社,1996:245-347
[37]Thomashow M F.So what's new in the field of plant cold acclimation? Lots![J].Plant Physiol.,2001,125(1):89
[38]Liu Q,Kasuga M,Sakuma Y.et al.Two transcription factors,DREB1 and DREB2,with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought and low-temperature-pesponsive gene expression,pespectively,in Arabidopsis[J].Plant Cell,1998,10:1391-1046
[39]简令成.植物冻害和抗冻性的细胞生物学研究[J].植物生理生化进展,1987,(5):1-16
[40]秦峰,李洁,张贵友,等.玉米中与DRE组件结合的转录因子的克隆和结构分析[J].植物学报,2003,45(3):331-339
[41]Dubouzet J G,Sakuma Y,Ito Y,et a.l.OsDREB genes in rice,Oryza sativa L.,encode transcription activators that function in drought,high salt and cold-responsive gene expression[J].Plant Journal,2003,33(4):751-763
[42]Sakuma Y,Lin Q,Dubouzet J G,et al.DNA binding specificity of the ERF/AP2 domain of Arabidopsis DREBs,transcription factors involved in dehydration and cold-inducible gene expression[J].Bio-chem.Biophys.Res.Commun.,2002,290(3):998-1009.
[43]Shinozaki K,Yamaguchi-Shinozaki K.Molecular responses to dehydration and low temperature:differences and crosstalk between two stress signaling pathways[J].Current Opinion in Plant Biology,2000,3:217-223
[44]Qin F,Sakuma Y,Li J,et al.Cloning and functional analysis of a novel DREBI/CBF transcription factor involved in cold-responsive gene expression in Zea mays L[J].Plant and Cell Physiology,2004,45(8):1042-1052
[45]Seki M,Kamei A,Yamaguchi-Shinozaki K,et al.Molecular responses to drought,salinity and frost:common and different paths for plant protection[j].Current Opinion in Biotechnology,2003,14:194-199
[46]Seki M,Narusaka M,Abe H,et al.Monitoring the expression pattern of 1300 Arabidopsis genes under drought and cold stresses by using a full-length cDNA microarray[J].Plant Cell,2001,13(1): 61-72
[47]Seki M,Narusaka M,Yamaguchi-Shinozaki K,et al.Arabidopsis encyclopedia using full-length cDNAs and its application[J].Plant Physiology and Bioche mistry,2001,39:211-220
[48]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[J].Plant Journal,1998,16:433-443
[49]Chinnusamy V,Ohta M,Kanrar S,et al.ICE1:a regulator of cold induced transcriptome and freezing tolerance in Arabidopsis[J].Genes & Evelopment,2003,17:1043-1054
[50]Ishitani M,Xiong L M,Lee H J,et al.HOS1,a genetic locus involved in cold responsive gene expression in Arabidopsis[J].Plant Cell,1998,10:1151-1161
[51]Lee H J,Xiong L M,Gong Z Z,et al.The Arabidopsis HOS1 gene negatively regulates cold signal transduction and encodes a RING finger protein that displays cold-regulated nucleo-cytoplasmic partitioning[J].Canes & Development,2001,15:912-924
[52]Novillo F,Alonso J M,Ecker J R,et al.CBF2/DREB1C is a negative regulator of CBF1/DREB1B and CBF3/DREB1A expression and plays a central role in stress tolerance in Arabidopsis[J].Proc.Natl.Acad.Sci.USA.,2004,101(11):3985-3990
[53]Fowler S G,Cook D,Thomashow M F.Low temperature induction of Arabidopsis CBF1,2,and 3 is gated by the circadian clock[J].Plant Physiol,2005,137(3):961-968
[54]Stockinger E J,Gilmour S J,ThomashowM F.Arabidopsis thaliana CBF1 encodes an AP2domain-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[J].Proc.Natl.Acad.Sci.USA.,1997,94:1035-1040
[55]曹琴,孔维府,温鹏飞.植物抗寒及其基因表达研究进展[J].生态学报,2004,4(24):806-810
[56]Lin Q,Knsuga M,Sakuma Y,et al.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[J].Plant Cell,1998,10:1391-1406
[57]Sakuma Y,Liu Q,Dubouzet J G,et al.DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs,transcription factors involved in dehydration and cold-inducible gene expression[J].Biochem.Biophys.Res.Commun.,2002,290:998-1009
[58]QIN Feng,LI Jie,ZHANG Gui-You,et al.Isolation and structural analysis of DRE-binding transcription factor from maize(Zea mays L.)[J].Acta.Botanica.,2003,45(3):331-339
[59]Dubouzet J G,Sakuma Y,Ito Y,et al.OsDREB genes in rice,Oryza sativa L.,encode transcription activators that function in drought,high-salt and cold responsive gene expression[J].Plant Journal,2003,33(4):751-763
[60]Jaglo K R,Kleff S,Amundsen K L,et al.Components of the Arabidopsis C-repeat/dehydrat -ion-responsive element binding factor cold-response pathway are conserved in Brassica napus and other plant species[J].Plant Physiol,2001,127:910-917
[61]Tang M,Lu S,Jing Y,et al.Isolation and identification of a cold-inducible gene encoding a putative DRE-binding transcription factor from Festucaa rundinacea[J].Plant.Physiol.Bio.chem.,2005,43:233-239
[62]Zhang J Y,Broeckling C D,Blancaflor E B,et al.Overexpression of WXP1,a putative Medicago truncatula AP2 domain-containing transcription factor gane,increases cuticular wax accumulation and enhances drought tolerance in transgenic alfalfa(Medicago saliva)[J].Plant J.,2005:1-19
[63]Novillo F,Alonso J M,Ecker J R,et al.CBF2/DREB1C is a negative regulator of CBF1/DREB1B and CBF3/DREB1A expression and plays a central role in stress tolerance in Arabidopsis[J].Proc.Natl.Acad.Sci.USA.,2004,101:3985-3990
[64]张开春,张军科.马哈利樱桃PGIP基因克隆及全序列测定[J].农业生物技术学报,2000,8(3):281-283
[65]李广平,乔玉山,陶建敏,等.中国李PGIP基因的克隆及序列分析[J].西北植物学报,2006,26(9):1870-1893
[66]李广平,房经贵,蔡斌华,等.梅PGIP基因的克隆及全序列分析[J].园艺学报,2006,33(1):125-127
[67]王家福,朱春林,陈桂信,等.龙眼梅PGIP基因的克隆及序列分析[J].果树学报,2007,24(1):55-58
[68]叶霞,陶建敏,蔡斌华,等.苹果铁结合蛋白基因的克隆及表达特性[J].园艺学报,2006,33(5):1051-1054.
[69]戚金亮.苹果铁高效基因型生物技术的研究—MxNrampl和MxIrtl基因的克隆[D].北京:中国农业大学硕士学位论文,2003
[70]任小林,金志强,李嘉瑞,等.猕猴桃钙调蛋白cDNA的克隆及测序[J].西北农业大学学报,1997,25(3):1-5
[71]Yu X C,Li M J,Gao G F,et al.Abscisic acid stimulates a calcium-dependent protein kinase in grape berry[J].Plant Physiology,2006,140:558-579
[72]邹克琴.苹果钙依赖蛋白激酶MdCPK1:分子克隆、表达、定位及被脱落酸的激活[D].北京:中国农业大学博士学位论文,2005
[72]王关林,杨怀义,夏然,等.草莓果实膜联蛋白基因(annfaf)全长cDNA克隆及序列分析[J].植 物学报,2001,43(8):873-875
[73]王俊英,张开春,王俊平,等.樱桃ACC氧化酶基因的克隆和序列分析[J].华北农报,2002,16(1):11-15
[74]Lu W J,Nakano R,Kubo Y,et al.Cloning and expression analysis of an XET cDNA in the peel and pulp of banana fruit ripening and softening[J].Acta.Botanica.Sinica.,2004,46(3):355-362
[75]韩继成,方宣钧.编码荔枝乙烯受体(LcERS1)的cDNA基因的克隆与分析[J].分子植物育种,2003,3(1):351-356
[76]周盛梅,宿红艳,王磊,等.苹果APETALA2同源基因的克隆和转化研究[J].园艺学报,2006,33(2):239-243
[77]Cao Q F,Wada M,Meng Y P,et al.Molecular cloning and expression analysis of a zinc finger protein gene in apple[J].Aeta.Botanica.Sinica.,2004,46(9):1091-1099
[78]宿红艳,周盛梅,王磊,等.草莓APETALA2同源基因的克隆及表达分析[J].西北植物学报,2005,25(10):1937-1942
[79]吴凡,徐勇,常凤启,等.桃中两个MADS-box基因的克隆与表达分析[J].遗传学报,2004,31(9):908-918
[80]丁燕,韩振海,许雪峰,等.柿花发育相关的MADS-box基因克隆与表达[J].园艺学报,2007,34(1):39-42
[81]郑学勤,李明芳.无核荔枝花发育相关MADS-box基因的克隆及结构分析[J].生物技术,2005,15(3):6-9
[82]刘莹,徐继忠,邵建柱,等.苹果矮化砧木P59离体叶片高效再生体系的建立[J].河北农业大学学报,2006,29(3):11-21
[83]陆玉英,阮经宙,林仁恭,等.柑桔生物快繁技术研究[J].中国农学通报,2006,22(11):79-82
[84]刘翠琼,汤浩茹,罗娅.不同基因型梨叶片离体培养和植株再生[J].园艺学报,2005,32(6):1080-1083
[85]秦永华,张上隆,徐凯,等.‘丰香'草莓叶片高效再生体系的建立[J].园艺学报,2005,32(1):101-104
[86]周厚成,罗静,赵霞,等.不同培养条件对幸香草莓离体叶片再生的影响[J].果树学报,2007,24(1):105-108
[87]李曜东,魏玉凝.肥城桃组培苗诱导、基因转化及其增值[J].果树学报,2003,20(1):70-72
[88]王关林,夏秀英,钟文田,等.樱桃砧木叶片再生系统建立及抗菌肽基因转化[J].园艺学报,2003,30(2):209-211
[89]王鸿,马锋旺,王发林,等.山杏下胚轴再生不定芽影响因子研究[J].果树学报,2005,22(4):399-40
[90]杨晓明,安黎哲,王雅梅,等.酿酒葡萄神索体胚发生及再生体系遗传稳定性分析[J].园艺学报,2006,33(6):1317-1320
[91]付志惠,李洪林,李琼,等.紫果猕猴桃幼胚愈伤组织诱导及植株再生[J].武汉植物学研究,2004,22(5):459-462
[92]曾继吾,易干军,张秋明.番木瓜体胚发生及植株再生研究[J].果树学报,2003,20(6):471-474
[93]赵慧,宋桂英.将菜豆几丁质酶基因导入苹果砧木的研究[J].激光生物学报,1998,7(3):163-167
[94]达克东,任德才,张松,等.超强表达介导豇豆胰蛋白酶抑制剂基因(CpT)转化苹果的研究[J].园艺学报,2001,28(1):57-58
[95]王关林,夏秀英,钟文田,等.樱桃砧木叶片再生系统建立及抗菌肽基因转化[J].园艺学报,2003,30(2):209-211
[96]刘庆忠,赵红军,刘鹏,等.抗菌肽MB39基因导入‘皇家嘎啦'苹果及其四倍体植株的培育[J].园艺学报,2001,28(5):392-398
[97]Finslad K,Marlin R R.Transformation of strawberry for virus ressitance[J].Aeta.Hort.,1995,385:86-90
[98]Norolli J,Aldwinekle H,Destefano B L.Transgenic Mailing 26 apple expressing the attacin E gene has increased resistance to Erwinia amylovora[J].Euphytica.,1993,77:123-128
[99]Vellieee G R,Ricci J C,Hemandez L,et al.Enhanced resistance to Botrytis cinerea mediated by the transgenic expression of the chitinase gene eh5B in strawberry[J].Transgenie Research,2006,15(1):57-68
[100]Mills D,Hammerschlag F A,Nordeen R O,el al.Evidence for the breakdown of cecropin B by proteinases in the intercellular fluid of peach leaves[J].Plant Sci.,1994,104:1722-1728
[101]马海军,冯学梅,马金平.果树生物技术育种进展[J].西北林学院学报,2006,21(4):93-95
[102]Graham J,Gordon S C,McNicol R J.The effect of the CpTi gene in strawberry against attack by vine weevil(Otiorhynchus suleatus F.Coleoptera:Curculionidae)[J].Annals of Applied Biology,1997,131:133-139
[103]Dandekar A M,blcGranaban G H,Vail P V,el al.High levels of expression of full-length cry IA(c) gene from Bacillus thuringiensis in transgenic walnut somatic embryos[J].Plant Sci.,1998,131(2):181-193
[104]Nehra NS,Stashnoff C.Direct shoot regeneration from strawberry leaf disks[J].J.Amer.Soc.Hort.Sci.,1989,114(6):1014-1018
[105]Nehra N S,Chibbar R N,Kartha K K,el al.Genetic transformation of strawberry by Agrobacterium tumefaciens using a leaf disk regeneration system[J].Plant Cell Rep.,1990,9(6): 293-298
[106]Nehra N S,Chibbar R N,Kartha K K,el al.Agrobacterium-mediated transformation of strawberry calli and recovery of transgenic plants[J].Plant Cell Rep.,1990,9(1):10-13
[107]James D J,Passey A J,Barbara DJ.Agrobacterium-mediated transformation of the cultivated strawberry(Fragaria×anannassa Duch) using disarmed binary vectors[J].Plant Sci.,1990,69:79-94
[108]El-Mansouri I,Mercado J A,Valpuesta V,el al.Shoot regeneration and Agrobacterium-mediated transformation of Fragariavesca L[J].Plant Cell Rep.,1996,15(8):642-646
[109]张志宏,吴禄平.草莓主栽品种Tudla遗传转化体系的建立[J].农业生物技术学报,1998,6(2):200-204
[110]Mathews H,Dewey V,Wagoner W,el al.Molecular and cellular evidence of chimaeric tissues in primary transgenic and elimination of chimaerism through improved selection protocols[J].Transgenic Research,1998,7(2):123-129
[111]Gruchaa A,Korbin M,Zurawicz E.Conditions of transformation and regeneration of Induka and Elista strawberry plants[J].Plant Cell Tiss.Org.Cult.,2004,79(2):153-160
[112]张红梅,王俊丽.‘全明星'草莓叶片遗传转化体系的建立[J].生物技术,2005,15(3):68-70
[113]Oosumi T,Gruszewski H A,Blischak L A,el al.High-efficiency transformation of the diploid strawberry(Fragaria vesca) for functional genomics[J].Planta,2006,223(6):1219-1230
[114]James D J,Passey A J,Esterbrook M A,el al.Progress in the introduction of transgenes for pest resistance in apples and strawberry[J].Phytoparasitica,1992,20(Suppl.):83-87
[115]Graham J,McNicol R J,Grieg K.Towards genetic based insert resistance in strawberry using the Cowpea trypin inhibitor gene[J].Ann.Appl.Bio.,1995,127(1):163-173.
[116]Graham J,Gordon S C,Smith K,el al.The effect of the Cowpea trypsin inhibitor in strawberry on damage by vine weevil under field conditions[J].J.Hort.Sci.Biotech.,2002,77(1):33-40.
[117]Finstad K,Martin RR.Transformation of strawberry for virus resistance[J].Acta.Hort.,1995,385:86-90.
[118]Martinelli L,Rugini E,Saccardo F.Genetic transformation for biotic stress resistance in horticultural plants[J].In vitro,1996,32(3):69-70
[119]Horowitz S,Freeman S,Sharon A.Use of green fluorescent protein-transgenic strains to study pathogenic and nonpathogenic lifestyles in Colletotrichum acutatum[J].Phytopathology,2002,92(7):743-749
[120]Asao H G,Nishizawa Y,Arai S,el al.Enhanced resistance against a fungal pathogen Sphaerotheca fumuli in transgenic strawberry expressing a rice chitinase gene[J].Plant Biotech.,1997,14(3),145-149
[121]Asao H,Shinmyo A,Nishizawa Y.Environmental risk evaluation of transgenic strawberry expressing a rice chitinase gene[J].Seibutsu-kogaku,2003,81:57-63
[122]Chalavi V,Tabaeizadeh Z,Thibodeau P.Enhanced resistance to Verticillium dahliae in transgenic strawberry plants expressing a Lycopersicon chilense ehitinase gene[J].J.Amer.Soc.Hort.Sci.,2003,128(5):747-753.
[123]Kim K K,Fravel D R,Papavizas G C.Identification of a metabolite produced by Talaromyces flavus as glucose oxidase and its role in biocontrol of Verticillium dahliae[J].Phytopathology,1988,78(4):488-492.
[124]金万梅,尹淑萍,鲁韧强,等.GO基因对草莓遗传转化及抗病性鉴定[J].分子植物育种,2005,3(6):797-800
[125]Schestibratov K A,Dolgov S V.Transgenic strawberry plants expressing a thaumatin Ⅱ gene demonstrate enhanced resistance to Botrytis cinema[J].Seientia.Horti.,2005,106:177-189
[126]秦永华.丰香草莓高效再生的机理及遗传转化研究[D].杭州:浙江大学博士学位论文,2005
[127]刘凤华,郭岩,郭东梅,等.转甜菜碱醛脱氢酶基因植物的耐盐性研究[J].遗传学报,1997,24(1):54-58
[128]王俊丽,葛会波,彭士琪,等.草莓外源LEAS基因的导入[J].园艺学报,2003,30(3):322-324
[129]WANG Jun-Li,GE Hui-Bo,PENG Shi-Qi,el al.Transformation of strawberry(Fragaria ananassa Duch.) with late embryogenesis abundant protein gene[J].J.Hort.Sci.and Biotech.,2004,79(5):735-738
[130]Firsov A P,Dolgov S V.Agrobacterium transformation and transfer of the antifreeze protein gene of winter flounder to the strawberry[J].Acta.Hort.,1999,484:581-586.
[131]Owens C L,Thomashow M F,Hancock J F,el al.CBF1 orthologs in sour cherry and strawberry and the heterologous expression of CBF1 in strawberry[J].J.Amer.Soc.Hort.Sci.,2002,127(4):489-494.
[132]Owens C L,Iezzoni A F,Hancock J F.Enhancement of freezing tolerance of strawberry by heterologous expression of CBF1[J].Acta.Hort.,2003,626:93-100
[133]Khammuang S,Dheeranupattana,Hanmuangjai P,el al.Agrobacterium-mediated transformation of modified antifreeze protein gene in strawberry[J].Songklanakarin J.Sci.Technol.,2005,27(4):693-703.
[134]Mario H,Sylvain D,Daniel N D,el al.Overexpression of the acidic dehydrin WCOR410 improves freezing tolerance in transgenic strawberry leaves[J].Plant Biotech.J.,2004,2(5):381-387
[135]Hennie J,Plessis DU,Brand RJ.Efficient genetic transformation of strawberry(Fragaria ananassa Duch.) cultivar Selekta[J].Acta.Hort.,1997,447:289-293
[136]张志宏,吴禄平,代红艳,等.草莓主栽品种再生和转化的研究[J].园艺学报,2001, 28(3):189-193.
[137]Morgan A,Baker C M,Lee K,el al.Production of herbicide tolerant strawberry through genetic engineering[J].Acta.Hort.,2002,567(11):113-115
[138]Bacbelier C,Graham J,Machray G,el al.Integration of an invertase gene to control sucrose metabolism in strawberry cultivars[J].Acta.Hort.,1997,436:161-163
[139]Mathews H,Wagoner W,Kellogg J,el al.Genetic transformation of strawberry:stable integration of a gene to control biosynthesis of ethylene[J].In vitro Cell Dev.Bio.,1995,31(1):36-43
[140]Jimenez-Bermudez S,Redondo-Nevado J,Munoz-Blanco J,el al.Manipulation of strawberry fruit softening by antisense expression of a pectate lyase gene[J].Plant Physiol,2002,128(2):751-759
[141]Al-Dlaigan Y,Cantliffe D,Klee H,el al.Highly reliable procedure for transformation and regeneration of the strawberry cultivar Sweet Charlie[J].Hort.Sci.,2003,38(4):725-729
[142]Al-Dlaigan Y I,Cantliffe D J,Klee H J.Protocols for gene transformation in strawberry(Fragaria ananassa cv.Sweet Charlie)[J].In vitro Cell Dev.Biol.,2002,38:109
[143]Fernanda A,Rocio G L,Jose L C,el al.Engineering increased vitamin C levels in plants by over-expression of a D-galacturonic acid reductase[J].Nature Biotech.,2003,21(2):177-181
[144]那杰,王关林,夏然,等.草莓annfaf基因反义融合表达载体构建及转基因植株的获得[J].中国农业科学,2006,39(3):582-586
[145]秦永华,张上隆.草莓转基因研究进展[J].遗传,2007,29(2):150-156
[146]吴雪梅,汤浩茹,文国琴.不同培养条件对丰香草莓离体叶片再生的影响[J].园艺学报,2004,31(5):657-659
[147]孙崇波,谢鸣,蒋桂华,等.草莓主栽品种丰香高效离体再生体系的研究[J].浙江农业学报,2003,15(2):69-72.
[148]牛建新,鲁晓燕,于艳华,等.外植体和培养因子对草莓不定芽诱导的影响[J].北方园艺,1999,3:30-31
[149]朱海生,潘东明,林义章,等.草莓高频离体再生体系的研究[J].西北植物学报,2007,2(5):0859-0863
[150]Thomas JC,Katteiman FR.Cytokininactivity induced by thidiazuxon[J].Plant Physiol,1986,81:681-683
[151]Passey AJ,Barrett K,James DJ.Adventitious shoot regeneration from seven commercial strawberry cultivars(Fragaria ananassa Duch) using a range of explant types[J].Plant Cell Reports,2003,21(5):397-401
[152]Qin YH,Zhang SL,Zhang LX.Regeneration mechanism of Toyonoka strawberry under different color plastic films[J].Plant Science,2005,168:1425-1431
[153]Nehra NS,Sthshnof C,Karthak K.Regeneration of plants from immature leaf-derived callus of strawberry(Frugaria ananassa Duch)[J].Plant Sci.Limerick,1990,66(1):119-126
[154]Yamaguchi-Shinozaki K,Shinozaki K,Arahidopsis DNA encoding two desiccation-responsive rd29 genes[J].Plant Physiol,1993,101:1119-1120
[155]袁维风,尹淑萍,金万梅等.根癌农杆菌介导转录因子CBF1基因对草莓的转化[J].生物学杂志,2006,23(4):37-40
[156]孙瑞芬.草莓再生及抗冻蛋白基因的转化.2004.中国优秀硕士论文期刊网
[157]Qiu-lin Qin,Jin-ge Liu.Isolation,optimization,and functional analysis of the cDNA encoding transcription factor OsDREB1B in Oryza Sativa L[J].Mol Breeding 6:1007-1032
[158]金建凤,高强,陈勇,等.转移拟南芥CBF1基因引起水稻植株脯氨酸含量提高[J].细胞生物学杂志,2005,27:73-76
[159]杨凤萍,梁荣奇,张立全,等.抗逆调节转录因子DREB1B基因转化多年生黑麦草的研究[J].西北植物学报,2006,26(7):1309-1315
[2]王小菁,李娘辉,潘瑞炽.植物生理学[M].北京:高等教育出版社,2006,432-476
[3]Weiser C J.Cold resistance and injury in woody plants[J].Science,1979,169:1269-1273
[4]Briggs D R.The chemistry of the living bark of the black locust tree in relation to frost hardiness[J].Arch.Biochem.,1949,23:8-11
[5]Hughes M A,Dunn M A.The molecular biology of plant acclimation to low temperature[J]Experimental Botany,1996,47(296):291-296
[6]Danyluk J,Houde M,Rassart E,et al.Differential expression of a gene encoding an acidic dehydrin in chilling sensitive and freezing-tolerant gramineae species[J].FEBS Letters,344,20-22
[7]Murata N,Yamaya J.Molecular species composition of phosphati-dylgylcerols from chilling semsitive and chilling resistance plants[J].Plant Cell Physiol,1983,24:81-84
[8]Guy C L,Haskell D,Neven L,et al.Llydratioirstate-resposive proteins link cold and droughtstress in spinach[J].Planta,1992,188:265-268
[9]Liu Qing,Kasuga M,Sakuma Y,et al.Two transcription factors,DREB1 and DREB2,with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought and low-temperatuxe-responsive gene expression respectively,in Arabidopsis[J].Plant Cell,1998,10(8):1391-1396
[10]Yamaguchi-Shinozaki K,Shinozaki K.A novel cis-acting element in an Arabidopsis Gene is involved in responsiveness to drought,low-temperature,or high-salt stress[J].Plant Cell,1994,6(2):251-264
[11]马建忠.植物的冷诱导基因[J].农业生物技术学报,1996,4(1):8-10
[12]余叔文,汤章城.植物生理与分子生物学[M].北京:科学出版社,1999,235-278
[13]Gibson S,Arondel V.Cloning of a temperature-regulated gene encoding a chloroplast-3 desaturase from Arabidopsis thaliana[J].Plant Physiol,1994,106:1615-1619
[14]Kodama H,Horiguchi G.Fatty acid desaturation during chilling acclimation is one of the factors involved in conferring low temperature tolerance to young tobacco leaves[J].Plant Physiology,1995,107(4):1177-1179
[15]Wada H,Gombos Z,Murata N.Enhancement of chilling tolerance of a cyanobacterium by genetic manipulaton of fatty acid desaturation[J].Nature,1990,347(13):200-205
[16]王艇,苏应娟,刘良式,等.植物低温诱导蛋白和低温诱导基因的表达调控[J].武汉植物学研究,1997,15(1):80-90
[17]Jarillo J A,Capei J,Leyva A,et al.Two related low-temperature-induced genes of Arabidopsis encode proteins showing high homology to 14-3-3 proteins,a family of putative Kinase regulators[J].Plant Molecular Biology,1994,25:693-696
[18]Lin C,Thomashow M F.A cold-regulated Arabidopsis gene encodes a polypetide having potent cryoprotective activity[J].Biochem.Biophysic.Res.Com..,1992,183:1103-1107
[19]江勇,贾士荣,费云标,等.抗冻蛋白及其在植物抗冻生理中的作用[J].植物学报,1999,7(41):678-182
[20]Antikainen M,Griffith M.Antifreeze protein accumulation in freezing tolerant cereals[J].Phy s-iologia Plantarum,1997,99:423-426
[21]Kasuga M,Liu Qi,Mura S.et al.Improving plant drought,salt,and freezing tolerance by gene transfer of a single stress- inducible transcription factor[J].Nature Biotechnologi,1999,17:287-293
[22]Kenward K D,Altschuler M,Hildebrand D.et al.Accumulation of type Ⅰ fish antifreeze protein in transgenic tobacco is cold-specific[J].Plant Molecular Biology,1993,23:377-382
[23]Guy L C.Altered gene expression during cold acclimation of spinach[J].Proc.Natl.Acad.Sci.USA,1985,82:3673-3677
[24]Autus N N.Constitutive expression of the cold-regulated Arabidopsis thsaliana cor15a gene affects both chloroplast feezing to tolerance[J].Proc.Natl.Acad.Sci.USA,1996,93:13404-1409
[25]Jaglo-Ottoseu K R.Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance[J].Sience,1998,280(5360):104-106
[26]Wallis J.Expression of synthetic antifreeze protein in potato reduces electrolyte release at freezing temperatures[J].Plant Molecular Biology,1997,35(3):323-330
[27]Culter A J.Winter flounder antifreeze protein improves the cold hardiness of plant tissue[J].Plant Physiol,1989,135(3):351-354.
[28]万巧兰.柑橘离体形态建成及转鱼类抗冻蛋白基因研究[D].北京:中国农业科学院硕士学位论文,1996
[29]Kenward K D.Type Ⅱ fish antifreeze protein accumulation in transgenic tobacco does not confer frost resistance[J].Transgenic Research,1999,8(2):105-117
[30]Georges F.Design and cloning of a synthetic gene for the flounder antifreeze protein and its expression in plant cell[J].Gene,1990,91:159-165
[31]黄永芬,汪清胤,付桂荣,等.美洲拟蝶抗冻蛋白基因(afps)导入西红柿的研究[J].生物化学杂志,1997,13(4):418-422
[32]Chakrabartty.Structure function relationships in a winter flounder antifreeze polypeptide[J].J.Biol.Chem.,1989,264(19):11307-11312
[33]Lee JS.The redaction of the freezing point of tobacco plants transformed with the gene encoding for the antifreeze protein from winter flounder[J].J.Cell.Biochem.,1990,14(supple):E303
[34]Griffith M.Antifreeze protein produced endogenously in winter rye leaves[J].Plant Physiol 1992,100:593-596
[35]Mckersis B D,Chen Y.Superoxide dismutase enbances tolerance of freezing stress in transgenic alfalfa(Medicago sative L.)[J].Plant Physiol,1993,103(4):1155
[36]赵微平.植物基因组构建、表达和调控[M].北京:首都师范大学出版社,1996:245-347
[37]Thomashow M F.So what's new in the field of plant cold acclimation? Lots![J].Plant Physiol.,2001,125(1):89
[38]Liu Q,Kasuga M,Sakuma Y.et al.Two transcription factors,DREB1 and DREB2,with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought and low-temperature-pesponsive gene expression,pespectively,in Arabidopsis[J].Plant Cell,1998,10:1391-1046
[39]简令成.植物冻害和抗冻性的细胞生物学研究[J].植物生理生化进展,1987,(5):1-16
[40]秦峰,李洁,张贵友,等.玉米中与DRE组件结合的转录因子的克隆和结构分析[J].植物学报,2003,45(3):331-339
[41]Dubouzet J G,Sakuma Y,Ito Y,et a.l.OsDREB genes in rice,Oryza sativa L.,encode transcription activators that function in drought,high salt and cold-responsive gene expression[J].Plant Journal,2003,33(4):751-763
[42]Sakuma Y,Lin Q,Dubouzet J G,et al.DNA binding specificity of the ERF/AP2 domain of Arabidopsis DREBs,transcription factors involved in dehydration and cold-inducible gene expression[J].Bio-chem.Biophys.Res.Commun.,2002,290(3):998-1009.
[43]Shinozaki K,Yamaguchi-Shinozaki K.Molecular responses to dehydration and low temperature:differences and crosstalk between two stress signaling pathways[J].Current Opinion in Plant Biology,2000,3:217-223
[44]Qin F,Sakuma Y,Li J,et al.Cloning and functional analysis of a novel DREBI/CBF transcription factor involved in cold-responsive gene expression in Zea mays L[J].Plant and Cell Physiology,2004,45(8):1042-1052
[45]Seki M,Kamei A,Yamaguchi-Shinozaki K,et al.Molecular responses to drought,salinity and frost:common and different paths for plant protection[j].Current Opinion in Biotechnology,2003,14:194-199
[46]Seki M,Narusaka M,Abe H,et al.Monitoring the expression pattern of 1300 Arabidopsis genes under drought and cold stresses by using a full-length cDNA microarray[J].Plant Cell,2001,13(1): 61-72
[47]Seki M,Narusaka M,Yamaguchi-Shinozaki K,et al.Arabidopsis encyclopedia using full-length cDNAs and its application[J].Plant Physiology and Bioche mistry,2001,39:211-220
[48]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[J].Plant Journal,1998,16:433-443
[49]Chinnusamy V,Ohta M,Kanrar S,et al.ICE1:a regulator of cold induced transcriptome and freezing tolerance in Arabidopsis[J].Genes & Evelopment,2003,17:1043-1054
[50]Ishitani M,Xiong L M,Lee H J,et al.HOS1,a genetic locus involved in cold responsive gene expression in Arabidopsis[J].Plant Cell,1998,10:1151-1161
[51]Lee H J,Xiong L M,Gong Z Z,et al.The Arabidopsis HOS1 gene negatively regulates cold signal transduction and encodes a RING finger protein that displays cold-regulated nucleo-cytoplasmic partitioning[J].Canes & Development,2001,15:912-924
[52]Novillo F,Alonso J M,Ecker J R,et al.CBF2/DREB1C is a negative regulator of CBF1/DREB1B and CBF3/DREB1A expression and plays a central role in stress tolerance in Arabidopsis[J].Proc.Natl.Acad.Sci.USA.,2004,101(11):3985-3990
[53]Fowler S G,Cook D,Thomashow M F.Low temperature induction of Arabidopsis CBF1,2,and 3 is gated by the circadian clock[J].Plant Physiol,2005,137(3):961-968
[54]Stockinger E J,Gilmour S J,ThomashowM F.Arabidopsis thaliana CBF1 encodes an AP2domain-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[J].Proc.Natl.Acad.Sci.USA.,1997,94:1035-1040
[55]曹琴,孔维府,温鹏飞.植物抗寒及其基因表达研究进展[J].生态学报,2004,4(24):806-810
[56]Lin Q,Knsuga M,Sakuma Y,et al.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[J].Plant Cell,1998,10:1391-1406
[57]Sakuma Y,Liu Q,Dubouzet J G,et al.DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs,transcription factors involved in dehydration and cold-inducible gene expression[J].Biochem.Biophys.Res.Commun.,2002,290:998-1009
[58]QIN Feng,LI Jie,ZHANG Gui-You,et al.Isolation and structural analysis of DRE-binding transcription factor from maize(Zea mays L.)[J].Acta.Botanica.,2003,45(3):331-339
[59]Dubouzet J G,Sakuma Y,Ito Y,et al.OsDREB genes in rice,Oryza sativa L.,encode transcription activators that function in drought,high-salt and cold responsive gene expression[J].Plant Journal,2003,33(4):751-763
[60]Jaglo K R,Kleff S,Amundsen K L,et al.Components of the Arabidopsis C-repeat/dehydrat -ion-responsive element binding factor cold-response pathway are conserved in Brassica napus and other plant species[J].Plant Physiol,2001,127:910-917
[61]Tang M,Lu S,Jing Y,et al.Isolation and identification of a cold-inducible gene encoding a putative DRE-binding transcription factor from Festucaa rundinacea[J].Plant.Physiol.Bio.chem.,2005,43:233-239
[62]Zhang J Y,Broeckling C D,Blancaflor E B,et al.Overexpression of WXP1,a putative Medicago truncatula AP2 domain-containing transcription factor gane,increases cuticular wax accumulation and enhances drought tolerance in transgenic alfalfa(Medicago saliva)[J].Plant J.,2005:1-19
[63]Novillo F,Alonso J M,Ecker J R,et al.CBF2/DREB1C is a negative regulator of CBF1/DREB1B and CBF3/DREB1A expression and plays a central role in stress tolerance in Arabidopsis[J].Proc.Natl.Acad.Sci.USA.,2004,101:3985-3990
[64]张开春,张军科.马哈利樱桃PGIP基因克隆及全序列测定[J].农业生物技术学报,2000,8(3):281-283
[65]李广平,乔玉山,陶建敏,等.中国李PGIP基因的克隆及序列分析[J].西北植物学报,2006,26(9):1870-1893
[66]李广平,房经贵,蔡斌华,等.梅PGIP基因的克隆及全序列分析[J].园艺学报,2006,33(1):125-127
[67]王家福,朱春林,陈桂信,等.龙眼梅PGIP基因的克隆及序列分析[J].果树学报,2007,24(1):55-58
[68]叶霞,陶建敏,蔡斌华,等.苹果铁结合蛋白基因的克隆及表达特性[J].园艺学报,2006,33(5):1051-1054.
[69]戚金亮.苹果铁高效基因型生物技术的研究—MxNrampl和MxIrtl基因的克隆[D].北京:中国农业大学硕士学位论文,2003
[70]任小林,金志强,李嘉瑞,等.猕猴桃钙调蛋白cDNA的克隆及测序[J].西北农业大学学报,1997,25(3):1-5
[71]Yu X C,Li M J,Gao G F,et al.Abscisic acid stimulates a calcium-dependent protein kinase in grape berry[J].Plant Physiology,2006,140:558-579
[72]邹克琴.苹果钙依赖蛋白激酶MdCPK1:分子克隆、表达、定位及被脱落酸的激活[D].北京:中国农业大学博士学位论文,2005
[72]王关林,杨怀义,夏然,等.草莓果实膜联蛋白基因(annfaf)全长cDNA克隆及序列分析[J].植 物学报,2001,43(8):873-875
[73]王俊英,张开春,王俊平,等.樱桃ACC氧化酶基因的克隆和序列分析[J].华北农报,2002,16(1):11-15
[74]Lu W J,Nakano R,Kubo Y,et al.Cloning and expression analysis of an XET cDNA in the peel and pulp of banana fruit ripening and softening[J].Acta.Botanica.Sinica.,2004,46(3):355-362
[75]韩继成,方宣钧.编码荔枝乙烯受体(LcERS1)的cDNA基因的克隆与分析[J].分子植物育种,2003,3(1):351-356
[76]周盛梅,宿红艳,王磊,等.苹果APETALA2同源基因的克隆和转化研究[J].园艺学报,2006,33(2):239-243
[77]Cao Q F,Wada M,Meng Y P,et al.Molecular cloning and expression analysis of a zinc finger protein gene in apple[J].Aeta.Botanica.Sinica.,2004,46(9):1091-1099
[78]宿红艳,周盛梅,王磊,等.草莓APETALA2同源基因的克隆及表达分析[J].西北植物学报,2005,25(10):1937-1942
[79]吴凡,徐勇,常凤启,等.桃中两个MADS-box基因的克隆与表达分析[J].遗传学报,2004,31(9):908-918
[80]丁燕,韩振海,许雪峰,等.柿花发育相关的MADS-box基因克隆与表达[J].园艺学报,2007,34(1):39-42
[81]郑学勤,李明芳.无核荔枝花发育相关MADS-box基因的克隆及结构分析[J].生物技术,2005,15(3):6-9
[82]刘莹,徐继忠,邵建柱,等.苹果矮化砧木P59离体叶片高效再生体系的建立[J].河北农业大学学报,2006,29(3):11-21
[83]陆玉英,阮经宙,林仁恭,等.柑桔生物快繁技术研究[J].中国农学通报,2006,22(11):79-82
[84]刘翠琼,汤浩茹,罗娅.不同基因型梨叶片离体培养和植株再生[J].园艺学报,2005,32(6):1080-1083
[85]秦永华,张上隆,徐凯,等.‘丰香'草莓叶片高效再生体系的建立[J].园艺学报,2005,32(1):101-104
[86]周厚成,罗静,赵霞,等.不同培养条件对幸香草莓离体叶片再生的影响[J].果树学报,2007,24(1):105-108
[87]李曜东,魏玉凝.肥城桃组培苗诱导、基因转化及其增值[J].果树学报,2003,20(1):70-72
[88]王关林,夏秀英,钟文田,等.樱桃砧木叶片再生系统建立及抗菌肽基因转化[J].园艺学报,2003,30(2):209-211
[89]王鸿,马锋旺,王发林,等.山杏下胚轴再生不定芽影响因子研究[J].果树学报,2005,22(4):399-40
[90]杨晓明,安黎哲,王雅梅,等.酿酒葡萄神索体胚发生及再生体系遗传稳定性分析[J].园艺学报,2006,33(6):1317-1320
[91]付志惠,李洪林,李琼,等.紫果猕猴桃幼胚愈伤组织诱导及植株再生[J].武汉植物学研究,2004,22(5):459-462
[92]曾继吾,易干军,张秋明.番木瓜体胚发生及植株再生研究[J].果树学报,2003,20(6):471-474
[93]赵慧,宋桂英.将菜豆几丁质酶基因导入苹果砧木的研究[J].激光生物学报,1998,7(3):163-167
[94]达克东,任德才,张松,等.超强表达介导豇豆胰蛋白酶抑制剂基因(CpT)转化苹果的研究[J].园艺学报,2001,28(1):57-58
[95]王关林,夏秀英,钟文田,等.樱桃砧木叶片再生系统建立及抗菌肽基因转化[J].园艺学报,2003,30(2):209-211
[96]刘庆忠,赵红军,刘鹏,等.抗菌肽MB39基因导入‘皇家嘎啦'苹果及其四倍体植株的培育[J].园艺学报,2001,28(5):392-398
[97]Finslad K,Marlin R R.Transformation of strawberry for virus ressitance[J].Aeta.Hort.,1995,385:86-90
[98]Norolli J,Aldwinekle H,Destefano B L.Transgenic Mailing 26 apple expressing the attacin E gene has increased resistance to Erwinia amylovora[J].Euphytica.,1993,77:123-128
[99]Vellieee G R,Ricci J C,Hemandez L,et al.Enhanced resistance to Botrytis cinerea mediated by the transgenic expression of the chitinase gene eh5B in strawberry[J].Transgenie Research,2006,15(1):57-68
[100]Mills D,Hammerschlag F A,Nordeen R O,el al.Evidence for the breakdown of cecropin B by proteinases in the intercellular fluid of peach leaves[J].Plant Sci.,1994,104:1722-1728
[101]马海军,冯学梅,马金平.果树生物技术育种进展[J].西北林学院学报,2006,21(4):93-95
[102]Graham J,Gordon S C,McNicol R J.The effect of the CpTi gene in strawberry against attack by vine weevil(Otiorhynchus suleatus F.Coleoptera:Curculionidae)[J].Annals of Applied Biology,1997,131:133-139
[103]Dandekar A M,blcGranaban G H,Vail P V,el al.High levels of expression of full-length cry IA(c) gene from Bacillus thuringiensis in transgenic walnut somatic embryos[J].Plant Sci.,1998,131(2):181-193
[104]Nehra NS,Stashnoff C.Direct shoot regeneration from strawberry leaf disks[J].J.Amer.Soc.Hort.Sci.,1989,114(6):1014-1018
[105]Nehra N S,Chibbar R N,Kartha K K,el al.Genetic transformation of strawberry by Agrobacterium tumefaciens using a leaf disk regeneration system[J].Plant Cell Rep.,1990,9(6): 293-298
[106]Nehra N S,Chibbar R N,Kartha K K,el al.Agrobacterium-mediated transformation of strawberry calli and recovery of transgenic plants[J].Plant Cell Rep.,1990,9(1):10-13
[107]James D J,Passey A J,Barbara DJ.Agrobacterium-mediated transformation of the cultivated strawberry(Fragaria×anannassa Duch) using disarmed binary vectors[J].Plant Sci.,1990,69:79-94
[108]El-Mansouri I,Mercado J A,Valpuesta V,el al.Shoot regeneration and Agrobacterium-mediated transformation of Fragariavesca L[J].Plant Cell Rep.,1996,15(8):642-646
[109]张志宏,吴禄平.草莓主栽品种Tudla遗传转化体系的建立[J].农业生物技术学报,1998,6(2):200-204
[110]Mathews H,Dewey V,Wagoner W,el al.Molecular and cellular evidence of chimaeric tissues in primary transgenic and elimination of chimaerism through improved selection protocols[J].Transgenic Research,1998,7(2):123-129
[111]Gruchaa A,Korbin M,Zurawicz E.Conditions of transformation and regeneration of Induka and Elista strawberry plants[J].Plant Cell Tiss.Org.Cult.,2004,79(2):153-160
[112]张红梅,王俊丽.‘全明星'草莓叶片遗传转化体系的建立[J].生物技术,2005,15(3):68-70
[113]Oosumi T,Gruszewski H A,Blischak L A,el al.High-efficiency transformation of the diploid strawberry(Fragaria vesca) for functional genomics[J].Planta,2006,223(6):1219-1230
[114]James D J,Passey A J,Esterbrook M A,el al.Progress in the introduction of transgenes for pest resistance in apples and strawberry[J].Phytoparasitica,1992,20(Suppl.):83-87
[115]Graham J,McNicol R J,Grieg K.Towards genetic based insert resistance in strawberry using the Cowpea trypin inhibitor gene[J].Ann.Appl.Bio.,1995,127(1):163-173.
[116]Graham J,Gordon S C,Smith K,el al.The effect of the Cowpea trypsin inhibitor in strawberry on damage by vine weevil under field conditions[J].J.Hort.Sci.Biotech.,2002,77(1):33-40.
[117]Finstad K,Martin RR.Transformation of strawberry for virus resistance[J].Acta.Hort.,1995,385:86-90.
[118]Martinelli L,Rugini E,Saccardo F.Genetic transformation for biotic stress resistance in horticultural plants[J].In vitro,1996,32(3):69-70
[119]Horowitz S,Freeman S,Sharon A.Use of green fluorescent protein-transgenic strains to study pathogenic and nonpathogenic lifestyles in Colletotrichum acutatum[J].Phytopathology,2002,92(7):743-749
[120]Asao H G,Nishizawa Y,Arai S,el al.Enhanced resistance against a fungal pathogen Sphaerotheca fumuli in transgenic strawberry expressing a rice chitinase gene[J].Plant Biotech.,1997,14(3),145-149
[121]Asao H,Shinmyo A,Nishizawa Y.Environmental risk evaluation of transgenic strawberry expressing a rice chitinase gene[J].Seibutsu-kogaku,2003,81:57-63
[122]Chalavi V,Tabaeizadeh Z,Thibodeau P.Enhanced resistance to Verticillium dahliae in transgenic strawberry plants expressing a Lycopersicon chilense ehitinase gene[J].J.Amer.Soc.Hort.Sci.,2003,128(5):747-753.
[123]Kim K K,Fravel D R,Papavizas G C.Identification of a metabolite produced by Talaromyces flavus as glucose oxidase and its role in biocontrol of Verticillium dahliae[J].Phytopathology,1988,78(4):488-492.
[124]金万梅,尹淑萍,鲁韧强,等.GO基因对草莓遗传转化及抗病性鉴定[J].分子植物育种,2005,3(6):797-800
[125]Schestibratov K A,Dolgov S V.Transgenic strawberry plants expressing a thaumatin Ⅱ gene demonstrate enhanced resistance to Botrytis cinema[J].Seientia.Horti.,2005,106:177-189
[126]秦永华.丰香草莓高效再生的机理及遗传转化研究[D].杭州:浙江大学博士学位论文,2005
[127]刘凤华,郭岩,郭东梅,等.转甜菜碱醛脱氢酶基因植物的耐盐性研究[J].遗传学报,1997,24(1):54-58
[128]王俊丽,葛会波,彭士琪,等.草莓外源LEAS基因的导入[J].园艺学报,2003,30(3):322-324
[129]WANG Jun-Li,GE Hui-Bo,PENG Shi-Qi,el al.Transformation of strawberry(Fragaria ananassa Duch.) with late embryogenesis abundant protein gene[J].J.Hort.Sci.and Biotech.,2004,79(5):735-738
[130]Firsov A P,Dolgov S V.Agrobacterium transformation and transfer of the antifreeze protein gene of winter flounder to the strawberry[J].Acta.Hort.,1999,484:581-586.
[131]Owens C L,Thomashow M F,Hancock J F,el al.CBF1 orthologs in sour cherry and strawberry and the heterologous expression of CBF1 in strawberry[J].J.Amer.Soc.Hort.Sci.,2002,127(4):489-494.
[132]Owens C L,Iezzoni A F,Hancock J F.Enhancement of freezing tolerance of strawberry by heterologous expression of CBF1[J].Acta.Hort.,2003,626:93-100
[133]Khammuang S,Dheeranupattana,Hanmuangjai P,el al.Agrobacterium-mediated transformation of modified antifreeze protein gene in strawberry[J].Songklanakarin J.Sci.Technol.,2005,27(4):693-703.
[134]Mario H,Sylvain D,Daniel N D,el al.Overexpression of the acidic dehydrin WCOR410 improves freezing tolerance in transgenic strawberry leaves[J].Plant Biotech.J.,2004,2(5):381-387
[135]Hennie J,Plessis DU,Brand RJ.Efficient genetic transformation of strawberry(Fragaria ananassa Duch.) cultivar Selekta[J].Acta.Hort.,1997,447:289-293
[136]张志宏,吴禄平,代红艳,等.草莓主栽品种再生和转化的研究[J].园艺学报,2001, 28(3):189-193.
[137]Morgan A,Baker C M,Lee K,el al.Production of herbicide tolerant strawberry through genetic engineering[J].Acta.Hort.,2002,567(11):113-115
[138]Bacbelier C,Graham J,Machray G,el al.Integration of an invertase gene to control sucrose metabolism in strawberry cultivars[J].Acta.Hort.,1997,436:161-163
[139]Mathews H,Wagoner W,Kellogg J,el al.Genetic transformation of strawberry:stable integration of a gene to control biosynthesis of ethylene[J].In vitro Cell Dev.Bio.,1995,31(1):36-43
[140]Jimenez-Bermudez S,Redondo-Nevado J,Munoz-Blanco J,el al.Manipulation of strawberry fruit softening by antisense expression of a pectate lyase gene[J].Plant Physiol,2002,128(2):751-759
[141]Al-Dlaigan Y,Cantliffe D,Klee H,el al.Highly reliable procedure for transformation and regeneration of the strawberry cultivar Sweet Charlie[J].Hort.Sci.,2003,38(4):725-729
[142]Al-Dlaigan Y I,Cantliffe D J,Klee H J.Protocols for gene transformation in strawberry(Fragaria ananassa cv.Sweet Charlie)[J].In vitro Cell Dev.Biol.,2002,38:109
[143]Fernanda A,Rocio G L,Jose L C,el al.Engineering increased vitamin C levels in plants by over-expression of a D-galacturonic acid reductase[J].Nature Biotech.,2003,21(2):177-181
[144]那杰,王关林,夏然,等.草莓annfaf基因反义融合表达载体构建及转基因植株的获得[J].中国农业科学,2006,39(3):582-586
[145]秦永华,张上隆.草莓转基因研究进展[J].遗传,2007,29(2):150-156
[146]吴雪梅,汤浩茹,文国琴.不同培养条件对丰香草莓离体叶片再生的影响[J].园艺学报,2004,31(5):657-659
[147]孙崇波,谢鸣,蒋桂华,等.草莓主栽品种丰香高效离体再生体系的研究[J].浙江农业学报,2003,15(2):69-72.
[148]牛建新,鲁晓燕,于艳华,等.外植体和培养因子对草莓不定芽诱导的影响[J].北方园艺,1999,3:30-31
[149]朱海生,潘东明,林义章,等.草莓高频离体再生体系的研究[J].西北植物学报,2007,2(5):0859-0863
[150]Thomas JC,Katteiman FR.Cytokininactivity induced by thidiazuxon[J].Plant Physiol,1986,81:681-683
[151]Passey AJ,Barrett K,James DJ.Adventitious shoot regeneration from seven commercial strawberry cultivars(Fragaria ananassa Duch) using a range of explant types[J].Plant Cell Reports,2003,21(5):397-401
[152]Qin YH,Zhang SL,Zhang LX.Regeneration mechanism of Toyonoka strawberry under different color plastic films[J].Plant Science,2005,168:1425-1431
[153]Nehra NS,Sthshnof C,Karthak K.Regeneration of plants from immature leaf-derived callus of strawberry(Frugaria ananassa Duch)[J].Plant Sci.Limerick,1990,66(1):119-126
[154]Yamaguchi-Shinozaki K,Shinozaki K,Arahidopsis DNA encoding two desiccation-responsive rd29 genes[J].Plant Physiol,1993,101:1119-1120
[155]袁维风,尹淑萍,金万梅等.根癌农杆菌介导转录因子CBF1基因对草莓的转化[J].生物学杂志,2006,23(4):37-40
[156]孙瑞芬.草莓再生及抗冻蛋白基因的转化.2004.中国优秀硕士论文期刊网
[157]Qiu-lin Qin,Jin-ge Liu.Isolation,optimization,and functional analysis of the cDNA encoding transcription factor OsDREB1B in Oryza Sativa L[J].Mol Breeding 6:1007-1032
[158]金建凤,高强,陈勇,等.转移拟南芥CBF1基因引起水稻植株脯氨酸含量提高[J].细胞生物学杂志,2005,27:73-76
[159]杨凤萍,梁荣奇,张立全,等.抗逆调节转录因子DREB1B基因转化多年生黑麦草的研究[J].西北植物学报,2006,26(7):1309-1315