甘蔗胁迫诱导表达基因ScCBF1的功能分析

Functional analysis of ScCBF1 in the response to stress in sugarcane

以甘蔗苗为材料研究了甘蔗CBF1转录激活因子ScCBF1在水杨酸(SA)、茉莉酸甲酯(Me JA)以及重金属镉(Cd Cl_2)和铜(Cu Cl_2)胁迫下的表达情况;将ScCBF1的原核表达载体p GEX-6P-1-ScCBF1转化大肠杆菌(Escherichia coli)Rosetta(DE3),检测其在NaCl、PEG8000胁迫下的生长情况.结果表明:ScCBF1在Me JA、SA、Cd Cl_2以及Cu Cl_2逆境胁迫下表达下调;在500 mmol·L-1NaCl胁迫下,含有重组质粒的细胞生长速度显著减缓,不同浓度NaCl胁迫下的平板胁迫结果进一步说明了ScCBF1基因不具耐盐性;在15%PEG8000胁迫下,含有重组质粒细胞的生长速度明显高于对照,不同浓度PEG8000胁迫下的平板胁迫结果说明了ScCBF1基因在应答干旱胁迫中具有抗逆性.构建了ScCBF1的植物表达载体p CAMBIA1301-ScCBF1并通过农杆菌侵染在本氏烟叶片中瞬时表达,在4℃低温胁迫下,T0代烟草植株长势显著优于对照.

To investigate the functions of ScCBF1 in alleviating salicylic acid（ SA）,methyl jasmonate（ Me JA）,heavy metal cadmium（ Cd） and copper（ Cu） stress,prokaryotic expression vector p GEX-6P-1-ScCBF1 was transformed into Rosetta（ DE3） strain（ Escherichia coli） in sugarcane seedlings,then the growth rate of transformed cell lines was investigated under NaCl and PEG8000 stress. The results showed that ScCBF1 was significantly down-regulated under the treatments of SA,Me JA,CdCl2 or CuCl2,respectively. With 500 mmol·L^-1NaCl,growth rates of cells containing the recombinant plasmid significantly slowed down. Further spot assay under different concentrations of NaCl confirmed that ScCBF1 was susceptible to NaCl. The growth of Rosetta/p GEX-6P-1-ScCBF1 cells was faster in LB liquid medium containing 15% PEG8000 compared to that of the control. Spot assay also revealed that ScCBF1 was resistant to drought stress. Moreover,the over expression vector of p CAMBIA-1301-ScCBF1 was constructed successfully and then was transformed into the leaves of Nicotiana benthamiana by Agrobacterium-mediated method. Transient expression of ScCBF1 in the tobacco plants confirmed that T0 generation plants with expressed ScCBF1 was more resistant to cold than the control under 4 ℃.