摘要
为了研究盐地碱蓬Ss DREB基因在转基因烟草中的功能,探究转基因烟草的抗逆生理机制,我们构建了SsDREB基因的植物高效表达载体p CAMBIA2301-Ss DREB,并通过农杆菌介导法将其导入烟草NC89中,通过Kan抗性、PCR及RT-PCR筛选鉴定阳性苗;对转基因烟草苗进行高盐、干旱等抗逆性分析,同时测定在不同浓度Na Cl和PEG6000处理后转基因烟草叶片的净光合速率(Pn)、气孔导度(Gs)、光能转化效率(Fv/Fm)、实际光量子效率(φPSⅡ)以及脯氨酸和可溶性糖的含量。结果表明:通过Kan抗性、PCR及RT-PCR筛选鉴定,最终获得12个株系转基因阳性苗。转基因植株的抗逆性与对照相比明显提高。在不同浓度Na Cl和PEG6000处理下,对照和转基因苗的Pn和Gs随着处理浓度的提高均呈现逐渐下降趋势,但在相同浓度处理下转基因苗的Pn和Gs均比对照高;随着Na Cl处理浓度的增大,对照和转基因植株的Fv/Fm和φPSⅡ均逐渐降低,但转基因苗的降低幅度比对照小;随着PEG处理浓度的提高,对照和转基因植株的Fv/Fm逐渐降低,但转基因苗的降低程度较对照小,而φPSⅡ下降趋势基本相同。在不同浓度Na Cl和PEG处理后,转基因植株和对照植株脯氨酸和可溶性糖含量均随着Na Cl处理浓度的增加而提高,但是转基因植株较对照植株增高更快。
To study the function of Ss DREB gene in transgenic tobacco and analyze physiological mechanisms of transgenic tobacco. In this study,the Ss DREB gene driven by constitutive promoter Ca MV 35 S was transformed into tobacco NC89 with Agrobaeterium EHA105 containing the plant expression vectors p CAMBIA2301-Ss DREB. The positive tobacco plants were selected by Kan resistance,PCR and RT-PCR. Drought and high salt tolerance of transgenic tobacco seedlings was determined,and net photosynthetic rate（ Pn）,stomatal conductance（ Gs）,the energy conversion efficiency（ Fv / Fm）,the actual photochemical efficiency（ φPSⅡ）,and the content of proline and soluble sugar of leaves of transgenic tobacco treated with different concentrations of Na Cl and PEG6000 respectively were measured. The results showed that 12 lines of positive transgenic seedlings were finally identified by Kan resistance,PCR and RT-PCR. The results of stress tolerance experiments showed that the overexpression of Ss DREB could improve resistance of transgenetic tobacco to abiotic stress. Under Na Cl and PEG6000 treatments with different concentrations,the Pn and Gs in control and transgenic plants decreased gradually with the increase of the treatment concentration,however,at the same treatment concentration,the Pn and Gs were higher in transgenic plants than that in control plants. With the increase of the treatment concentration of Na Cl,Fv / Fm and φPSⅡ in the control and the transgenic plants declined gradually,but those of the transgenic plants decreased more slowly than those of con-trol plants. Under different concentration of PEG treatment,Fv / Fm of control and transgenic plants gradually decreased,with the increase of the PEG concentration but the decrease of transgenic plant Fv / Fm was slower when compared with control plants; However,the φPSⅡdid not show obvious differences in changes between the transgenic and control plants. Proline and soluble sugar contents of the transgenic and control plants increased with the increase of the treatment concentration of Na Cl and PEG,but proline and soluble sugar contents in the transgenic plants were higher than those in control plants.
出处
《华北农学报》
CSCD
北大核心
2014年第6期155-162,共8页
Acta Agriculturae Boreali-Sinica
基金
江苏省盐土生物资源研究重点实验室开放课题项目(JKLBS 2012004)
