Analysis of Physiological Mechanism of Stress Tolerance in SsDREB Transgenic Tobacco

doi:10.7668/hbnxb.2014.06.027

Abstract

Abstract: 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 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 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 improve resistance of transgenetic tobacco to abiotic stress.Under NaCl 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 NaCl,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 control 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 NaCl and PEG,but proline and soluble sugar contents in the transgenic plants were higher than those in control plants.

Key words: Tobacco, SsDREB gene, Transgene, Stress tolerance, Physiological index

CLC Number:

S572.01

SUN Xiao-bo, JIA Xin-ping, LIU Xiao-xue, MA Hong-xiang, YE Xiao-qing. Analysis of Physiological Mechanism of Stress Tolerance in SsDREB Transgenic Tobacco[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(6): 155-162. doi: 10.7668/hbnxb.2014.06.027.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2014.06.027

http://www.hbnxb.net/EN/Y2014/V29/I6/155

References

[1] Barrels D,Sunkar R.Drought and salt tolerance in plants [J].Critical Reviews in Plant Sciences,2005,24:23-58.
[2] Yamaguchi-Shinozaki K,Shinozaki K.Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses[J].Annual Review of Plant Biology,2006,57:781-803.
[3] 李杰,李晶,朱延明,等. DREB1A 基因植物表达载体的构建[J].东北农业大学学报,2003,34(2):199-204.
[4] Nj1 S,Lourenco T,oliveira M M.Transcription factors and regulation of photosynthetic and related metabolism under environmental stresses[J].Annals Botany,2009,103(4):609-623.
[5] Nakashima K,Ito Y,Yamaguchi-Shinozaki K.Transcriptional regulatory networks in response to abiotic stresses in Arabidopsis and grasses[J].Plant Physiology,2009,149(1):88-95.
[6] Magnani E,Sjolander K,Hake S.From endonucleases to transcription factors:Evolution of the AP2 DNA binding domain in plants[J].Plant Cell,2004,16(9):2265-2277.
[7] Sakuma Y,Liu Q,Dubouzet J G,et al.DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs,transcription factors involved in dehydration-and cold-inducible gene expression[J].Biochemical and Biophysical Research Communications,2002,290(3):998-1009.
[8] 赵艳红,郎明林,王小龙,等.纤毛鹅观草DREB类基因 RcDREB1 的克隆及其蛋白结合干旱应答元件DRE的功能验证[J].植物遗传资源学报,2013,14(5):857-863.
[9] 张梅,刘炜,毕玉平.植物中DREBs类转录因子及其在非生物胁迫中的作用[J].遗传,2009,31(3):236-244.
[10] Xu Z S,Chen M,Li L C,et al.Functions and application of the AP2/ERF transcription factor family in crop improvement[J].Journal of Integrative Plant Biology,2011,53(7):570-585.
[11] Zhang X X,Tang Y J,Ma Q B,et al.OsDREB2A,a rice transcription factor,significantly affects salt tolerance in transgenic soybean[J].PLOS One,2013,8(12):e83011.
[13] Wang B S,Luttge U,Ratajczak R.Specific regulation of SOD isoforms by NaCl and osmotic stress in leaves of the C-3 halophyte Suaeda salsa L.[J].Journal of Plant Physiology,2004,161(3):285-293.
[14] Song J,Fan H,Zhao Y Y,et al.Effect of salinity on germination,seedling emergence,seedling growth and ion accumulation of a euhalophyte Suaeda salsa in an intertidal zone and on saline inland[J].Aquatic Botany,2008,88(4):331-337.
[15] 黄玮,李志刚,乔海龙,等.旱盐互作对盐地碱蓬生长及其渗透调节物质的影响[J].中国生态农业学报,2008,16(1):173-178.
[16] 刘晓雪,孙晓波,王秀娥,等.盐地碱蓬 SsDREB 基因的克隆与表达分析研究[J].核农学报,2011,25(4):684-691.
[17] 邹琦.植物生理生化实验指导[M].北京:中国农业出版社,1995:41-46.
[18] Irigoyen J J,Einerich D W,Sánchez-Díaz M.Water stress induced changes in concentrations of proline and total soluble sugars in nodulated alfalfa(Medicago sativd)plants [J].Physiologia Plantarum,1992,84:55-60.
[19] 张志良.植物生理学实验指导[M].北京:高等教育出版社,2002:127-128.
[20] 陈丽萍,何道一.植物抗旱耐盐基因的研究进展[J].基因组学与应用生物学,2010,29(3):542-549.
[21] 王小华,庄南生.脯氨酸与植物抗寒性的研究进展[J].中国农学通报,2008,24(11):398-402.
[22] 张怀山,赵桂琴,栗孟飞,等.中型狼尾草幼苗对PEG、低温和盐胁迫的生理应答[J].草业学报,2014,23(2):180-188.
[23] Liu Q,Kasuga M,Sakuma Y,et al.Two transcription factors, DREB1 and DREB2,with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought-and low-temperature-responsive gene expression,respectively,in Arabidopsis[J].Plant Cell,1998,10(8):1391-1406.
[24] Chen M,Wang Q Y,Cheng X G,et al.GmDREB2,a soybean DRE-binding transcription factor,conferred drought and high-salt tolerance in transgenic plants[J].Biochemical and Biophysical Research Communications,2007,353(2):299-305.
[25] Lata C,Prasad M.Role of DREBs in regulation of abiotic stress responses in plants[J].Journal of Experimental Botany,2011,62(14):4731-4748.
[26] Agarwal P K,Agarwal P,Reddy M K,et al.Role of DREB transcription factors in abiotic and biotic stress tolerance in plants[J].Plant Cell Reports,2006,25(12):1263-1274.
[27] Matsukura S,Mizoi J,Yoshida D,et al.Comprehensive analysis of rice DREB2-type genes that encode transcription factors involved in the expression of abiotic stress-responsive genes[J].Molecular Genetics and Genomics:MGG,2010,283(2):185-196.
[28] Sun J W,Peng X J,Fan W H,et al.Functional analysis of BpDREB2 gene involved in salt and drought response from a woody plant Broussonetia papyrifera [J].Gene,2014,535(2):140-149.
[29] Qin F,Kakimoto M,Sakuma Y,et al.Regulation and functional analysis of ZmDREB2A in response to drought and heat stresses in Zea mays L.[J].Plant Journal,2007,50(1):54-69.
[30] Wang Q,Guan Y,Wu Y,et al.Over expression of a rice OsDREB1F gene increases salt,drought,and low temperature tolerance in both Arabidopsis and rice[J].Plant Molecular Biology,2008,67(6):589-602.
[31] Chen M,Xu Z,Xia L,et al.Cold-induced modulation and functional analyses of the DRE-binding transcription factor gene, GmDREB3,in soybean(Glycine max L.)[J].Journal of Experimental Botany,2009,60(1):121-135.
[32] Savitch L V,Allard G,Seki M,et al.The effect of overexpression of two Brassica CBF/DREB1-like transcription factors on photosynthetic capacity and freezing tolerance in Brassica napus [J].Plant and Cell Physiology,2005,46(9):1525-1539.
[33] Oh S J,Kwon C W,Choi D W,et al.Expression of barley HvCBF4 enhances tolerance to abiotic stress in transgenic rice[J].Plant Biotechnology Journal,2007,5(5):646-656.
[34] Igarashi Y,Yoshiba Y,Sanada Y,et al.Characterization of the gene for deltalpyrroline-5-carboxylate synthetase and correlation between the expression of the gene and salt tolerance in Oryza sativa L[J].Plant Molecular Biology,1997,33(5):857-865.
[35] Costa F G,Pham Thi A T,Pimentel C,et al.Differences in growth and water relations among Phaseolus vulgaris cultivars in response to induced drought stress[J].Environmental and Experimental Botany,2000,43(3):227-237.
[36] 刘晓东,李洋洋,何淼.PEG模拟干旱胁迫对玉带草生理特性的影响[J].草业科学,2012,29(5):687-693.
[37] 谢虹,杨兰,李忠光.脯氨酸在植物非生物胁迫耐性形成中的作用[J].生物技术通报,2011(2):23-27,60.
[38] 宗俊梅.冰凌花转录因子 AaDREB1 基因的表达分析及功能鉴定[D].长春:吉林农业大学,2011.
[39] 陈浩东,罗伯祥,陈芬,等. CbDREB1A 基因表达载体构建及转化水稻光温敏核不育系的研究[J].杂交水稻,2009,24(6):49-53,76.
[40] Zhao J S,Ren W,Zhi D Y,et al. Arabidopsis DREB1A/CBF3 bestowed transgenic tall fescue increased tolerance to drought stress[J].Plant Cell Reports,2007,26(9):1521-1528.

[1]	LI Ban, LÜ Ying, YANG Mingxuan, SONG Ting, YU Fang, LIU Zhiwen. *Effects of Saline-alkali Stress on Physiology and Molecular Mechanism of Brassica napus* L.** [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(3): 86-93.
[2]	HE Jing, DONG Jianxin, CONG Ping, SONG Wenjing, MA Xiaogang, GUAN Ensen, WANG Dahai. Rapid Improvement of Maize Straw Carbon Form on Soil Organic Carbon and Comprehensive Fertility in Tobacco Planting Soil [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(2): 132-141.
[3]	CHEN Xiangyu, LI Yanhua, LIU Zhike. Effects of TSINGKE Water Retaining Agent on External Morphology and Physiological Indexes of Epipremnum aureum [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(S1): 165-169.
[4]	CHEN Wentao, PAN Gen, TANG Huijuan, CHANG Li, LI Defang, ZHAO Lining, CHEN Anguo, LI Jianjun. *Cloning,Expression and Functional Analysis of HcMS1* Gene from Kenaf** [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 64-74.
[5]	HE Mingyu, XIAO Hanqian, DENG Xiaohua, LI Liangyong, LI Wujin, ZHOU Fumei, CHEN Zhifeng, XIAO Yansong, HUANG Qionghui, HUANG Jie. Growth and Dry Matter Accumulation and Nutrient Utilization Efficiency of Rich Flavor Style Paddy-tobacco [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 139-146.
[6]	PENG Guangjue, WANG Zhiyong, HU Tong, ZHENG Zhongyi, TANG Chungui, LIU Yongjun, LI Qiang, ZHOU Qingming, LI Juan. Effects of Deep Vertical Rotary Tillage on Soil Physical Characteristics and Root Development of Flue-Cured Tobacco in Paddy-Tobacco Growing Area in Changsha [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(1): 134-142.
[7]	WANG Peng, ZHENG Xuebo, LIANG Hongbo, SONG Wenjing, JI Xuan, XU Yanli, KUANG Shuai, DONG Jianxin. Effects of Different Fertilization Models on Active Organic Carbon Components and Enzyme Activities of Tobacco-growing Brown Soil [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(1): 187-196.
[8]	JIANG Dan, WEN Huan, PAN Shanshan, LAN Shiqi, NAN Wenbin, LIANG Yongshu, ZHANG Hanma, QIN Xiaojian. Analysis of Grain Type and Physiological Characteristics for a Large Grain Rice [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 126-132.
[9]	Lü Lihua, LI Qian, LEI Mingshuai, YAO Yanrong, JIA Xiuling. Analysis of Physiological Characteristics of Heat Resistant Wheat [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 138-144.
[10]	SUN Shu, ZHANG Xiaoqi, ZHAO Le, ZHOU Zhenfeng. Effects of Dibutyl Phthalate and Monobutyl Phthalate on Wheat Seed Germination and Protective Enzyme Activity of Seedling [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 145-151.
[11]	HUANG Ziyuan, WANG Tianye, WANG Chengyu, BAI Yichen, WU Yangsheng, LIU Shuxia. Effects of Different Fertilization Treatments on Soil Physical and Chemical Properties, Photosynthetic Characteristics and Yield of Maize in Semi-arid Area [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 234-240.
[12]	GAO Yukun, YANG Puyuan, XIANG Xiaodong, WEI Shilin, REN Genzeng, YIN Congpei, LIANG Hongkai, CUI Jianghui, CHANG Jinhua. Response of Different Salt Tolerant Sorghum Varieties to Salt Stress in the Whole Growth Period [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(6): 113-121.
[13]	XIE Bing, ZHAN Xiaoxu, LUO Yanhong, LIU Binxiang, KONG Fanlei, YUAN Jichao. Transcriptome Analysis of the Effect of Different Rootstocks on the Resistance of Grafted Flue-cured Tobacco to Bacterial Wilt [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(4): 177-186.
[14]	HE Fumeng, LI Xiuyu, ZHAO Xiaocan, WU Jiawen, ZHU Yuanfang, ZHOU Lei, SHI Qihai, LIU Di, LI Fenglan. *Analysis of Disease Resistance and Physiological Characteristics of Transgenic Tobacco with Solanum tuberosum StPR1* Gene** [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(3): 184-192.
[15]	WANG Haibo, WANG Xiaodi, HAN Xiao, XIE Jimeng, SHI Xiangbin, JI Xiaohao, WANG Baoliang, ZHENG Xiaocui, WANG Zhiqiang, LIU Fengzhi. Evaluation of Environmental Adaptability of 22 Grape Cultivars with Different Evaluation Methods [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(S1): 92-98.

Please choose a citation manager

Content to export