DREB(dehydration responsive element binding)转录因子通过调控下游多个抗逆相关基因的表达,能有效提高植物的抗逆性。将构建的植物高效表达载体GmDREB::pCAMBIA1304,借助优化的floral-dip法,转入模式植物拟南芥,并经潮霉素Hygromycin...DREB(dehydration responsive element binding)转录因子通过调控下游多个抗逆相关基因的表达,能有效提高植物的抗逆性。将构建的植物高效表达载体GmDREB::pCAMBIA1304,借助优化的floral-dip法,转入模式植物拟南芥,并经潮霉素Hygromycine(40～50mg.L-1)抗性筛选得到22棵抗性植株。对抗性植株再进行PCR和GUS检测获得19颗阳性苗,阳性率为86.3%。对T1代种子进行抗性分离比例统计,有4个株系的分离比例接近3:1,符合孟德尔遗传定律,说明外源基因GmDREB在这些株系的染色体中可能是单拷贝插入。继续对上述4个株系的后代进行抗性筛选,现已得到2个纯合的转基因株系。导入的报告基因GUS组织染色检测表明,转入大豆DREB基因在拟南芥的根系和子叶中均有大量表达,并在叶脉中表达。展开更多
The transcription factor dehydration-responsive element binding protein(DREB)is able to improve tolerance to abiotic stress in plants by regulating the expression of downstream genes involved in environmental stress r...The transcription factor dehydration-responsive element binding protein(DREB)is able to improve tolerance to abiotic stress in plants by regulating the expression of downstream genes involved in environmental stress resistance.The objectives of this study were to evaluate the salt tolerance of GmDREB1 transgenic wheat(Triticum aestivum L.)and to evaluate its physiological and protein responses to salt stress.Compared with the wild type,the transgenic lines overexpressing GmDREB1 showed longer coleoptiles and radicles and a greater radicle number at the germination stage,as well as greater root length,fresh weight,and tiller number per plant at the seedling stage.The yield-related traits of transgenic lines were also improved compared with the wild type,indicating enhanced salt tolerance in transgenic lines overexpressing GmDREB1.Proteomics analysis revealed that osmotic-and oxidative-stressrelated proteins were up-regulated in transgenic wheat leaves under salt stress conditions.Transgenic wheat had higher levels of proline and betaine and lower levels of malondialdehyde and relative electrolyte leakage than the wild type.These results suggest that GmDREB1 regulates the expression of osmotic-and oxidative-stress-related proteins that reduce the occurrence of cell injury caused by high salinity,thus improving the salt tolerance of transgenic wheat.展开更多
Under stress conditions such as droughthigh-salinity and low-temperature, the transcription factorof DREB (dehydration responsive element binding proteins)improved efficiently stress resistance by regulating the ex-pr...Under stress conditions such as droughthigh-salinity and low-temperature, the transcription factorof DREB (dehydration responsive element binding proteins)improved efficiently stress resistance by regulating the ex-pression of its downstream genes with various environmentastress resistance in plants. GmDREB gene (GenBank Acces-sion No. AF514908) encoding a stress-inducible transcriptionfactor was cloned by screening a cDNA library of Glycinemax cv. Jinong 27 with yeast one-hybrid method. GmDREBgene was 910 bp in length and encoded 174 amino acids con-taining a conserved AP2/EREBP DNA-binding domain of 58amino acids. Two conserved functional amino acids, valineand glutamic acid, were located on the 14th and the 19thamino acid residues in the conserved structural domain. Analkaline amino acid region (KKR) related to a nuclear local-ization signal was at the N-terminal, while an acidic aminoacid region (DDD) related to trans-activation was at theC-terminal. Plant expression vectors were constructed andtransformed into wheat by bombardment. In total, 13 trans-genic plants with Ubi::GmDREB and 11 transgenic plantswith rd29A::GmDREB were identified from 103 regenerationplants by molecular analysis. The drought and salt tolerancesof T1 transgenic lines with Ubi::GmDREB orrd29A::GmDREB were demonstrated to be improved ascompared to wild type. The result also suggested that bothUbiquitin and rd29A promoters could effectively drive theexpression of the GmDREB gene and enhance drought andsalt tolerance of T1 plants.展开更多
文摘DREB(dehydration responsive element binding)转录因子通过调控下游多个抗逆相关基因的表达,能有效提高植物的抗逆性。将构建的植物高效表达载体GmDREB::pCAMBIA1304,借助优化的floral-dip法,转入模式植物拟南芥,并经潮霉素Hygromycine(40～50mg.L-1)抗性筛选得到22棵抗性植株。对抗性植株再进行PCR和GUS检测获得19颗阳性苗,阳性率为86.3%。对T1代种子进行抗性分离比例统计,有4个株系的分离比例接近3:1,符合孟德尔遗传定律,说明外源基因GmDREB在这些株系的染色体中可能是单拷贝插入。继续对上述4个株系的后代进行抗性筛选,现已得到2个纯合的转基因株系。导入的报告基因GUS组织染色检测表明,转入大豆DREB基因在拟南芥的根系和子叶中均有大量表达,并在叶脉中表达。
基金supported by the National Transgenic Key Project from the Ministry of Agriculture of China(2014ZX08011-003)the Agricultural Science and Technology Innovation Program(ASTIP)
文摘The transcription factor dehydration-responsive element binding protein(DREB)is able to improve tolerance to abiotic stress in plants by regulating the expression of downstream genes involved in environmental stress resistance.The objectives of this study were to evaluate the salt tolerance of GmDREB1 transgenic wheat(Triticum aestivum L.)and to evaluate its physiological and protein responses to salt stress.Compared with the wild type,the transgenic lines overexpressing GmDREB1 showed longer coleoptiles and radicles and a greater radicle number at the germination stage,as well as greater root length,fresh weight,and tiller number per plant at the seedling stage.The yield-related traits of transgenic lines were also improved compared with the wild type,indicating enhanced salt tolerance in transgenic lines overexpressing GmDREB1.Proteomics analysis revealed that osmotic-and oxidative-stressrelated proteins were up-regulated in transgenic wheat leaves under salt stress conditions.Transgenic wheat had higher levels of proline and betaine and lower levels of malondialdehyde and relative electrolyte leakage than the wild type.These results suggest that GmDREB1 regulates the expression of osmotic-and oxidative-stress-related proteins that reduce the occurrence of cell injury caused by high salinity,thus improving the salt tolerance of transgenic wheat.
基金This work was supported by the National 863 Project(Grant No.2002AA224081)National Special Project for Plant Transgenic and Industry(Grant No.JY03-A-18).
文摘Under stress conditions such as droughthigh-salinity and low-temperature, the transcription factorof DREB (dehydration responsive element binding proteins)improved efficiently stress resistance by regulating the ex-pression of its downstream genes with various environmentastress resistance in plants. GmDREB gene (GenBank Acces-sion No. AF514908) encoding a stress-inducible transcriptionfactor was cloned by screening a cDNA library of Glycinemax cv. Jinong 27 with yeast one-hybrid method. GmDREBgene was 910 bp in length and encoded 174 amino acids con-taining a conserved AP2/EREBP DNA-binding domain of 58amino acids. Two conserved functional amino acids, valineand glutamic acid, were located on the 14th and the 19thamino acid residues in the conserved structural domain. Analkaline amino acid region (KKR) related to a nuclear local-ization signal was at the N-terminal, while an acidic aminoacid region (DDD) related to trans-activation was at theC-terminal. Plant expression vectors were constructed andtransformed into wheat by bombardment. In total, 13 trans-genic plants with Ubi::GmDREB and 11 transgenic plantswith rd29A::GmDREB were identified from 103 regenerationplants by molecular analysis. The drought and salt tolerancesof T1 transgenic lines with Ubi::GmDREB orrd29A::GmDREB were demonstrated to be improved ascompared to wild type. The result also suggested that bothUbiquitin and rd29A promoters could effectively drive theexpression of the GmDREB gene and enhance drought andsalt tolerance of T1 plants.
