James Victoria A, Neibaur Isaac, Altpeter Fredy
Agronomy Department, Plant Molecular and Cellular Biology Program, Genetics Institute, University of Florida, PO Box 110300, Gainesville, FL 32611, USA.
Transgenic Res. 2008 Feb;17(1):93-104. doi: 10.1007/s11248-007-9086-y. Epub 2007 Apr 6.
The dehydration-responsive element binding proteins (DREB1)/C-repeat (CRT) binding factors (CBF) function as transcription activators and bind to the DRE/CRT cis-acting element commonly present in the promoters of abiotic stress-regulated genes. A DREB1A transcription factor ortholog was isolated from a xeric, wild barley (Hordeum spontaneum L.) accession, originating from the Negev desert. Sequence comparison revealed a very high degree of sequence conservation of HsDREB1A to the published barley (Hordeum vulgare L.) DREB1A. Constitutive expression of the HsDREB1A gene was able to trans-activate a reporter gene under transcriptional control of the stress-inducible HVA1s and Dhn8 promoters. HsDREB1A was subcloned under transcriptional control of the stress-inducible barley HVA1s promoter and introduced into the apomictic bahiagrass (Paspalum notatum Flugge) cultivar 'Argentine'. HsDREB1A integration and stress inducible expression was detected in primary transgenic bahiagrass plants and apomictic seed progeny by Southern blot, RT-PCR and northern blot analysis respectively. Transgenic bahiagrass plants with stress-inducible expression of HsDREB1A survived severe salt stress and repeated cycles of severe dehydration stress under controlled environment conditions, in contrast to non-transgenic plants. The observed abiotic stress tolerance is very desirable in turf and forage grasses like bahiagrass, where seasonal droughts and irrigation restrictions affect establishment, persistence or productivity of this perennial crop.
脱水响应元件结合蛋白(DREB1)/C重复序列(CRT)结合因子(CBF)作为转录激活因子发挥作用,并与非生物胁迫调控基因启动子中常见的DRE/CRT顺式作用元件结合。从源自内盖夫沙漠的旱生野生大麦(Hordeum spontaneum L.)种质中分离出一个DREB1A转录因子直系同源物。序列比较显示,HsDREB1A与已发表的大麦(Hordeum vulgare L.)DREB1A具有高度的序列保守性。HsDREB1A基因的组成型表达能够在胁迫诱导型HVA1s和Dhn8启动子的转录控制下反式激活报告基因。将HsDREB1A亚克隆到胁迫诱导型大麦HVA1s启动子的转录控制下,并导入无融合生殖的巴哈雀稗(Paspalum notatum Flugge)品种‘阿根廷’中。分别通过Southern杂交、RT-PCR和Northern杂交分析,在转基因巴哈雀稗植株及其无融合生殖种子后代中检测到了HsDREB1A的整合及胁迫诱导型表达。与非转基因植株相比,在可控环境条件下,具有胁迫诱导型表达HsDREB1A的转基因巴哈雀稗植株在严重盐胁迫和反复的严重脱水胁迫循环中存活了下来。对于像巴哈雀稗这样的草坪草和饲草来说,所观察到的非生物胁迫耐受性是非常理想的,因为季节性干旱和灌溉限制会影响这种多年生作物的建植、持久性或生产力。