Study on Photosynthetic and Chlorophyll Fluorescence Characteristics of Transgenic Tobacco with AcCBF1 Gene

doi:10.6048/j.issn.1001-4330.2019.02.008

Abstract

Abstract: 【Objective】 To explore the effects of AcCBF1 gene on photosynthesis, and provide a scientific basis for breeding new varieties of almonds.【Method】 In this experiment, the transgenic tobacco with AcCBF1 gene was obtained by Agrobacterium-mediated method. After a week of continuous outdoor high temperature weather at 37 ℃, the diural variation characteristics of photosynthetic parameters and chlorophyll fluorescence parameters were measured and analyzed with the wild tobacco as control.【Result】 In this experiment, three transgenic tobaccos were successfully obtained by Agrobacterium-mediated method, and PCR analysis showed that the AcCBF1 gene had been stably integrated into the tobacco genome. Both Pn and Gs of transgenic tobacco and wild tobacco showed a "double peak" curve and peaks appeared at the same time. The first peak appeared at 12:00, The phenomenon of "mid-day depression" occurred at 16:00, and the second peak appeared at 18:00. The daily average values of Pn were 6.496 and 4.553 μ mol/(m²·s), respectively ; the daily average values of Gs were 157.357 and 133.371 μ mol/(m²·s) , respectively. The Fo of transgenic tobacco and wild tobacco showed a trend of increasing first and then decreasing, and Fm showed a trend of decreasing first and then increasing. The daily variation trends of Fv/Fm, Fv/Fo, and ΦPSII were the same, and they all showed a trend of decreasing first and then increasing. The daily average values of Fv/Fm values were 0.821 and 0.736, respectively. The daily average values of Fv/Fo were 5.044 and 3.391, respectively. The daily average values of ΦPSII were 0.763 and 0.600, respectively.【Conclusion】By analyzing and comparing the changes of photosynthetic parameters and fluorescence chlorophyll parameters, we found that transgenic AcCBF1 tobacco has higher assimilation capacity of CO₂, stronger photosynthetic ability, more stable photosynthetic reaction center and stronger adaptability to high temperature comparing with wild tobacco. To cope with drought and high temperature in summer, we could select and breed new almond varieties with AcCBF1 high-expression.

Key words: the AcCBF1 gene of almond; tobacco; photosynthetic parameters; chlorophyll fluorescence parameters

摘要： 【目的】扁桃AcCBF1基因对光合作用的影响,为扁桃新品种选育提供科学依据。【方法】采用农杆菌介导法获得转AcCBF1基因烟草,在经过连续室外37℃高温天气一周后,以野生烟草为对照,测定和分析试验烟草的光合参数和叶绿素荧光参数日变化特性。【结果】利用农杆菌介导法成功获得3株转基因烟草,经PCR检测证明AcCBF1基因已稳定地整合到烟草基因组中。转基因烟草和野生烟草的Pn、Gs均呈现“双峰”曲线,且峰值出现时间一致。12:00达到第一个峰值,16:00出现光合“午休”,18:00达到第二个峰值。二者的Pn日均值分别为6.496 、4.553 μmol/(m²·s);Gs日均值分别为157.357 、133.371 μmol/(m²·s)。转基因烟草和野生烟草的Fo均呈现出先上升后下降的趋势,Fm呈现先下降后上升的趋势。Fv/Fm、Fv/Fo、ΦPSII与Fm日变化趋势一致,均呈现先下降后上升的趋势,二者的Fv/Fm日均值分别为0.821、0.736;Fv/Fo日均值分别为5.044、3.391、ΦPSII日均值分别为0.763、0.600。【结论】与野生烟草相比,转AcCBF1基因烟草同化CO₂能力较高,光合能力较强,光合反应中心较稳定,对高温的适应能力较强。扁桃AcCBF1基因能够有效改善烟草的光合作用,维护光合反应中心的稳定;选育扁桃AcCBF1高表达品种应对夏季的干旱高温。

关键词: 扁桃AcCBF1基因, 烟草, 光合参数, 叶绿素荧光参数

CLC Number:

S662.9

ZHANG Qi, LI Peng, TIAN Jia, LI Jiang. Study on Photosynthetic and Chlorophyll Fluorescence Characteristics of Transgenic Tobacco with AcCBF1 Gene[J]. Xinjiang Agricultural Sciences, 2019, 56(2): 267-277.

张琦, 李鹏, 田嘉, 李疆. 转扁桃AcCBF1基因对烟草光合及叶绿素荧光特性的影响[J]. 新疆农业科学, 2019, 56(2): 267-277.

References 32

[1]	Martínez-Gómez, P., Sánchez-Pérez, R., Dicenta, F., Howad, W., Arús, P., & Gradziel, T. M. (2007). Almond. Fruits and Nuts.
[2]	田建保. 中国扁桃[M]. 北京: 中国农业出版社, 2008: 91-117.TIAN Jian-bao.(2008). Chinese Almond [M]. Beijing: China Agriculture Press:91-117.(in Chinese)
[3]	殷继英. 新疆扁桃品质相关指标测定分析[D]. 乌鲁木齐: 新疆农业大学硕士论文, 2015.YIN Ji-ying. (2015). Determination and Analysis of Related Indexes to Almond Quality in Xinjiang [D]. Master Dissertation. Xinjiang Agricultural University, Urumqi. (in Chinese)
[4]	李疆, 胡芳名, 李文胜, 等. 扁桃的栽培及研究概况[J]. 果树学报, 2002, 19(5): 346-350.LI Jiang, HU Fang-ming, LI Wen-sheng, et al. (2002). Advances of Almond Growing and Research [J]. Journal of Fruit Science, 19(5): 346-350. (in Chinese)
[5]	李疆. 中国果树科学与实践-阿月浑子、扁桃[M]. 西安: 陕西科学技术出版社, 2015:96-98.LI Jiang. (2015). The Science and Practice of Fruit trees in China-pistachio and Almond [M]. Xian: Shaanxi Science and Technology Press. (in Chinese)
[6]	USDA. (2017). 2017 California Almond Forecast. National Agricultural Statistics Service.
[7]	Lata, C., & Prasad, M. (2011). Role of drebs in regulation of abiotic stress responses in plants. Journal of Experimental Botany, 62(14), 4,731-4,748.
[8]	Agarwal, P. K., Agarwal, P., Reddy, M. K., & Sopory, S. K. (2006). Role of dreb transcription factors in abiotic and biotic stress tolerance in plants. Plant Cell Reports, 25(12):1,263-1,274.
[9]	胡亚杰. 干旱胁迫下烟草中DREB类转录因子过表达产生的生理效应研究初探[D]. 郑州: 河南农业大学硕士论文, 2008.HU Ya-jie. (2008). Preliminary study on physiological effects of DREB-Like Transcription Factor Overexperssion in tobacco under drought stress [D]. Master Dissertation. Henan Agricultural University, Zhenzhou. (in Chinese)
[10]	Sazegari, S. , Niazi, A. , & Ahmadi, F. S. . (2015). A study regulatory network with promoter analysis for arabidopsis dreb-genes. Bioinformation, 11(2):101-106. .
[11]	Aixin, L. , Mingqi, Z. , Donghui, W. , Hu, C. , Chenjiang, Y. , & Juan, L. . (2017). Transcriptome profiling reveals the negative regulation of multiple plant hormone signaling pathways elicited by overexpression of c-repeat binding factors. Frontiers in Plant Science, 8, 1647.
[12]	李宁. 新疆野扁桃AlsCBF基因的克隆及功能验证[D]. 乌鲁木齐: 新疆农业大学硕士论文, 2012.LI Ning. (2012). Cloning and Functional Verification ofAlsCBF Gene Amygdalus ledebouriana Schlecht in Xinjiang [D]. Master Dissertation. Xinjiang Agricultural University, Urumqi. (in Chinese)
[13]	张亮, 李疆, 帕提曼·阿布都热合曼, 等. 扁桃AcCBF1转录因子的克隆及表达分析[J]. 果树学报, 2015, 16(5): 763-768.ZHANG Liang, LI Jiang, PATIMAN Abudureheman, et al. (2015). Clone and Expression Analysis of AcCBF1 Gene from Almond [J]. Journal of Fruit Science, 16(5): 763-768. (in Chinese)
[14]	郭淑朋, 张亮, 李疆, 等. 扁桃AcCBF2基因的克隆及其在逆境胁迫下的表达分析[J]. 分子植物育种, 2017, 14(3): 814-820.GUO Shu-peng, ZHANG Liang, LI Jiang, et al. (2017). Molecular Cloning of AcCBF2 Gene and Its Expression in Almond [J]. Molecular Plant Breeding, 14(3):814-820. (in Chinese)
[15]	Dinakar, C., Djilianov, D., & Bartels, D. (2012). Photosynthesis in desiccation tolerant plants: energy metabolism and antioxidative stress defense. Plant Science, 182(1):29-41.
[16]	李鹏民, 高辉远, Strasser R J. 快速叶绿素荧光诱导动力学分析在光合作用研究中的应用[J]. 植物生理与分子生物学学报, 2005, 31(6): 559-566.LI Peng-min, GAO Hui-yuan, Strasser R J. (2005). Application of the Fast Chlorophyll Fluorescence Induction Dynamics Analysis in Photosynthesis Study [J]. Journal of Plant Physiology and molecular Biology, 31(6): 559-566. (in Chinese)
[17]	成敏敏, 陈柯伊, 朱雪玉, 等. 花叶矢竹复绿期光合特性及叶绿体结构[J]. 林业科学, 2018, 54(4): 1-10.CHENG Min-min, CHEN Ke-yi, ZHU Xue-yu, et al. (2018). Photosynthetic Characteristics and Chloroplast ultrastructure of Pseudosasa japonica f. akebonosuji during Green-Revertible Albino Stage [J]. Scientia Silvae Sinicae, 54(4): 1-10. (in Chinese)
[18]	Goto, N. , Miyazaki, H. , Nakamura, N. N. , Terai, H. , Ishida, N. , & Mitamura, O. . (2008). Relationships between electron transport rates determined by pulse amplitude modulated (pam) chlorophyll fluorescence and photosynthetic rates by traditional and common methods in natural freshwater phytoplankton. Fundamental and Applied Limnology / Archiv für Hydrobiologie, 172(2):121-134.
[19]	晁朝霞, 任艳萍, 钱进, 等. 新牧1号苜蓿两种抗逆相关启动子的功能分析[J]. 草业学报, 2017, 26(1): 131-141.CHAO Zhao-xia, REN Yan-ping, QIAN Jin, et al. (2017). Functional analysis of two stress-related promoters in Medicago varia cultivar Xin-mu No.1[J]. Acta Prataculture Sinica, 26(1): 131-141. (in Chinese)
[20]	Thomashow, M. F. . (1998). Role of cold-responsive genes in plant freezing tolerance. Plant Physiology, 118(1), 1-7.
[21]	韩拓,冯丽丽, 马婷, 等. 干旱绿洲植物叶片光合作用特性及季节变化规律[J]. 兰州大学学报(自然科学学报), 2016, 52(4): 492-497.HAN Tuo, FENG Li-li, MA Ting, et al. (2016). Characteristics and seasonal variations of plant leaf photosynthesis in arid oasis [J]. Journal of Lanzhou University (Natural Sciences Ed.) , 52(4): 492-497. (in Chinese)
[22]	牛莹莹, 廖康, 赵世荣, 等. 不同栽植密度库尔勒香梨光合及叶绿素荧光特性比较[J]. 经济林研究, 2017, 35(1): 8-14.NIU Ying-ying, LIAO Kang, ZHAO Shi-rong, et al. (2017). Comparisons of photosynthetic characteristics and chlorophyll fluorescence characteristics in Korla fragrant pear trees under different planting densities [J]. Nonwood Forest Research, 35(1):8-14. (in Chinese)
[23]	刘卫群, 胡亚杰, 甄焕菊, 等. 干旱胁迫对转BnDREB1-5基因烟草光合特性的影响[J]. 武汉植物学研究, 2008, 26(3):294-297.LIU Wei-qun, HU Ya-jie, ZHEN Huan-ju, et al. (2008). The Effect on Photosynthetic Characteristics of Transgenic Tobacco with BnDRER1-5 Gene by Drought Stress [J]. Journal of Wuhan Botanical Research, 26(3):294-297. (in Chinese)
[24]	马薇, 廖康, 牛莹莹, 等. 温室与露地栽培葡萄光合及荧光参数差异分析[J]. 新疆农业科学, 2016, 53(3): 393-398.MA Wei, LIAO Kang, NIU Ying-ying, et al. (2016). Analysis of Difference of Photosynthesis Characteristic and Fluorescence Parameters of Grape in Greenhouse and Open Field Cultivation [J]. Xinjiang Agricultural Sciences, 53(3):393-398. (in Chinese)
[25]	李月灵, 金则新. 玉兰叶片光合速率和叶绿素荧光参数的日变化[J]. 江苏农业科学, 2012. 40(9): 164-168.LI Yue-ling, JIN Ze-xin. (2012). Diurnal Changes of Photosynthetic Rate and Chlorophyll Fluorescence Parameters of Magnolia denudata Leaves [J]. Jiangsu Agricultural Sciences, 40(9): 164-168. (in Chinese)
[26]	钟敏, 张文标, 邹梁峰, 等. 高温下猕猴桃光合作用和叶绿素荧光特性的日变化[J]. 江西农业大学学报, 2018,40(3): 472-478.ZHONG Min, ZHANG Wen-biao, ZOU Liang-feng, et al. (2018). Diurnal Variation of Photosynthesis and Chlorophyll Fluore-Scence Characteristics in Kiwifruit Under High Temperature Condition [J]. Journal of Jiangxi Agricultural University, 40(3):472-478. (in Chinese)
[27]	Farquhar, G. D. F., & Sharkey, T. D. (1982). Stomatal conductance and photosynthesis. Annu.rev.plant Physiol, 33(1), 317-345.
[28]	陈建明, 俞晓平, 程家安. 叶绿素荧光动力学及其在植物抗逆生理研究中的应用[J]. 浙江农业学报, 2006, 18(1):51-55.CHEN Jian-ming, YU Xiao-ping, CHENG Jia-an. (2006). The application of chlorophyll fluorescence kinetics in the study of physiological responses of plants to environmental stresses [J]. Acta Agriulturae Zhejiangensis, 18(1): 51-55. (in Chinese)
[29]	赵会杰, 邹琦, 于振文. 叶绿素荧光分析技术及其在植物光合机理研究中的应用[J]. 河南农业大学学报, 2000,34(3): 248-251.ZHAO Hui-jie, ZOU Qi, YU Zhen-wen. (2000). Chlorophyll fluorescence analysis technique and its application to photosynthesis of plant [J] . Journal of Henan Agricultural University, 34(3): 248-251. (in Chinese)
[30]	张守仁. 叶绿素荧光动力学参数的意义及讨论[J]. 植物学通报, 1999, 16(4): 444-448.ZHANG Shou-ren. (1999). A Discussion on Chlorophyll Fluorescence Kinetics Parameters and Their Significance [J]. Chinese Bulletin of Botany, 16(4): 444-448. (in Chinese)
[31]	赵跃锋, 陈昆, 张清华. 盐胁迫对茄子光合特性、叶绿素荧光及保护酶活性的影响[J]. 山西农业科学, 2018, 46(11): 1797-1 799, 1 906.ZHAO Yue-feng, CHEN Kun, ZHANG Qing-hua. (2018). Effects of Salt Stress on Photosynthetic Characteristics, Chlorophyll Fluorescence and Protective Enzyme Activity of Eggplant [J]. Journal of Shanxi Agricultural Sciences, 46(11): 1,797-1,799, 1,906. (in Chinese)
[32]	刘建福. 红千层叶片光合速率和叶绿素荧光参数日变化[J]. 西南大学学报(自然科学版), 2007, 29(5): 93-100.LIU Jian-fu. (2007). Diurnal Changes of Photosynthetic Rate and Cholorophyll Fluorescence Parameters in Callistemon viminalis DC[J]. Journal of Southwest University(Natural Science Ed.), 29(5): 93-100. (in Chinese)