CAJ | 학술논문

为了研究盐地碱蓬SsDREB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Ss-DREB基因的植物高效表达载体pCAMBIA2301-SsDREB,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 NaCl 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NaCl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NaCl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NaCl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NaCl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。
위료연구염지감봉SsDREB기인재전기인연초중적공능,탐구전기인연초적항역생리궤제,아문구건료Ss-DREB기인적식물고효표체재체pCAMBIA2301-SsDREB,병통과농간균개도법장기도입연초NC89중,통과Kan항성、PCR급RT-PCR 사선감정양성묘；대전기인연초묘진행고염、간한등항역성분석,동시측정재불동농도 NaCl 화PEG6000처리후전기인연초협편적정광합속솔( Pn )、기공도도( Gs )、광능전화효솔( Fv/Fm )、실제광양자효솔(φPSⅡ)이급포안산화가용성당적함량。결과표명：통과Kan항성、PCR급RT-PCR사선감정,최종획득12개주계전기인양성묘。전기인식주적항역성여대조상비명현제고。재불동농도NaCl화PEG6000처리하,대조화전기인묘적Pn화Gs수착처리농도적제고균정현축점하강추세,단재상동농도처리하전기인묘적Pn화Gs균비대조고；수착NaCl처리농도적증대,대조화전기인식주적Fv/Fm화φPSⅡ균축점강저,단전기인묘적강저폭도비대조소；수착PEG처리농도적제고,대조화전기인식주적Fv/Fm축점강저,단전기인묘적강저정도교대조소,이φPSⅡ하강추세기본상동。재불동농도NaCl화PEG처리후,전기인식주화대조식주포안산화가용성당함량균수착NaCl처리농도적증가이제고,단시전기인식주교대조식주증고경쾌。
To study the function of SsDREB gene in transgenic tobacco and analyze physiological mechanisms of transgenic tobacco. In this study,the SsDREB gene driven by constitutive promoter CaMV 35S was transformed into tobacco NC89 with Agrobaeterium EHA105 containing the plant expression vectors pCAMBIA2301-SsDREB. The positive tobacco plants were selected by Kan resistance,PCR and RT-PCR. Drought and high salt tolerance of trans-genic 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 NaCl 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 SsDREB could im-prove resistance of transgenetic tobacco to abiotic stress. Under NaCl and PEG6000 treatments with different con-centrations,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 NaCl,Fv/Fm andφPSⅡin 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 de-creased,with the increase of the PEG concentration but the decrease of transgenic plant Fv/Fm was slower when compared with control plants;However,theφPSⅡdid not show obvious differences in changes between the transgen-ic and control plants. Proline and soluble sugar contents of the transgenic and control plants increased with the in-crease of the treatment concentration of NaCl and PEG, but proline and soluble sugar contents in the transgenic plants were higher than those in control plants.