欧美杨锌指蛋白转录因子基因（ZxZF）的遗传转化及抗旱性初步分析

doi:10.11707/j.1001-7488.20140305

Abstract

Abstract:

The C2H2 zinc finger protein transcription factor plays an important role in the activation of stress-related genes, and increase in plant resistance. In this study, an C2H2 zinc finger protein transcription factor (ZxZF) from shrub Sarcozygium xanthoxylon with strong drought tolerance was used as exogenous gene,and the Agrobacterium-mediated method was applied for transgenic transformation to an elite hybrid clone of Populus × euramericana cl.‘Bofeng1’. A total 80 resistant plants were gained via strict selection by kanamycin. Among them 17 positive transgenic plants were verified by PCR analysis, and five transgenic plants were finally confirmed with one copy of the exogenous gene by Southern blot analysis. Four of these clones were verified over-expression of ZxZF by RT-PCR and qRT-PCR analysis. Under the drought stress conditions, transgenic clones showed more than 10% significantly higher of relative water content (RWC) and accumulation of proline content, compared with non-transgenic control plants. These results showed an improved tolerance to drought stress in transgenic poplar. Our study suggested that ZxZF transcription factor played a crucial role in transgene poplar response to drought stress and was useful in developing trees with enhanced tolerance to drought.

Key words: Populus×euramericana, zinc-finger protein gene, genetic transformation, drought resistance

CLC Number:

Zhang Weixi, Liu Boyang, Ding Changjun, Zhang Bingyu, Huang Qinjun, Chu Yanguang, Su Xiaohua. Transformation of Zinc Finger Protein Transcription Factor Gene (ZxZF) and Preliminary Evaluation of Drought Tolerance in Populus × euramericana[J]. Scientia Silvae Sinicae, 2014, 50(3): 31-37.

References

崔旭东, 苏晓华, 张冰玉, 等. 2012. 欧美杨渤丰1号高效组培再生体系建立. 林业科学研究, 25(2): 157-162.
(Cui X D, Su X H, Zhang B Y, et al. 2012. Establishment of an efficient regeneration System of Populus × euramericana cl.'Bofeng 1'. Forest Research, 25(2): 157-162. [in Chinese] )
李合生. 2007. 植物生理生化实验原理和技术. 北京: 高等教育出版社, 164, 137-137, 258-260.
李义良, 苏晓华, 张冰玉, 等. 2007. 外源SacB基因在银腺杂种杨基因组中的表达及抗旱性分析. 北京林业大学学报, 29(2): 1-6.
(Li Y L, Su X H, Zhang B Y, et al. 2007. Molecular detection and drought tolerance of SacB-transgenic Populus alba×P. glandulosa. Journal of Beijing Forestry University, 29(2): 1-6. [in Chinese] )
苏晓华, 张冰玉,黄烈健, 等. 2003.转基因林木研究进展. 林业科学研究, 16(1): 95-103.
(Su X H, Zhang B Y, Huang L J, et al. 2003. Forest Research, Genetic engineering of forest trees. Forest Research, 16(1): 95-103. [in Chinese] )
萨姆布鲁克 J, 拉塞尔 D W. 2002. 分子克隆实验指南.3版. 北京:科学出版社.
(Sambrook J, Russell D W. 2002. Molecular cloning: A laboratory manual, 2nd Ed. Beijing: Science Press. [in Chinese] )
王俊英, 尹伟伦, 夏新莉. 2005. 胡杨锌指蛋白基因克隆及其结构分析. 遗传, 27(2): 245-248.
(Wang J Y, Yin W L, Xia L L. 2005. Cloning and structure analysis of zinc finger protein gene in Populus euphratica oliver. Hereditas(Beijing), 27(2): 245-248. [in Chinese] )
严安心, 谢瑜, 吉柔风, 等. 2012. 一个麻疯树RING型锌指蛋白基因JcRFP 1 的克隆及表达. 植物生理学报, 48(5): 456-464.
(Yan A X, Xie Y, Ji R F, et al. 2012. Cloning and expression of a RING finger protein gene JcRFP1 from Jatropha curcas L. Plant Physiology Journal, 48(5): 456-464. [in Chinese] )
杨金水, 王光清. 1995. 转基因的失活与沉默. 生物工程进展, 15 (3) :41-45.
张冰玉, 苏晓华, 黄秦军, 等. 2005. 转果聚糖蔗糖转移酶基因银腺杨的获得(英文). 林业科学, 41(3): 48-53.
(Zhang B Y, Su X H, Huang Q J, et al. 2005. Regeneration of transgenic poplar(Populus alba×P.glandulosa) expressing levansucrase from bacillus subtilis. Scientia Silvae Sinicae, 41(3): 48-53. [in Chinese] )
中国科学院中国植物志编辑委员会. 1998. 中国植物志(第43卷, 第一分册). 北京: 科学出版社, 140-142.
(Editorialcommittee of Flora of China. 1998. Flora reipublicae popularis sinicae(Vol.43 Book 1). Beijing: Science Press, 140-142. [in Chinese] )
朱家红, 李辉亮, 屠发志, 等. 2006. 巴西橡胶树环锌指蛋白基因克隆及其分子特性(英文). 植物生理与分子生物学学报, 32(6): 627-633.
(Zhu J H, Li H L, Tu F Z, et al. 2006. Cloning and molecular characterization of a RING Zinc-Finger Gene of Hevea brasiliensis. Journal of Plant Physiology and Molecular Biology, 32(6): 627-633. [in Chinese] )
An Y, Wang Y, Lou L, et al. 2011. A novel zinc-finger-like gene from Tamarix hispida is involved in salt and osmotic tolerance. J Plant Res, 124(6):689-97.
Castonguay Y, Markhart A H. 1992. Leaf gas exchange in water-stressed common bean and tepary bean. Crop Sci, 32(4): 980-986.
Davletova S, Schlauch K, Coutu J, et al. 2005. The zinc-finger protein Zat12 plays a central role in reactive oxygen and abiotic stress signaling in Arabidopsis. Plant Physiology, 139(2): 847-856.
Englbrecht C C, Schoof H, Böhm S. 2004. Conservation, diversification and expansion of C2H2 zinc finger proteins in the Arabidopsis thaliana genome. BMC Genomics, 5(1):39.
Hsieh T, Lee J, Charng Y, et al. 2002a. Tomato plants ectopically expressing Arabidopsis CBF 1 show enhanced resistance to water deficit stress. Plant Physiology, 130(2): 618-626.
Hsieh T, Lee J, Yang P, et al. 2002b. Heterology expression of the Arabidopsis C-Repeat/Dehydration Response Element Binding Factor 1 gene confers elevated tolerance to chilling and oxidative stresses in transgenic tomato. Plant Physiology, 129(3): 1086-1094.
Huang J, Sun S, Xu D, et al. 2009. Increased tolerance of rice to cold, drought and oxidative stresses mediated by the overexpression of a gene that encodes the zinc finger protein ZFP245. Biochemical and Biophysical Research Communications, 389(3): 556-561.
Jain M, Tyagi A K, Khurana J P. 2008. Constitutive expression of a meiotic recombination protein gene homolog, OsTOP6A1, from rice confers abiotic stress tolerance in transgenic Arabidopsis plants. Plant Cell Reports, 27(4): 767-778.
Kim J C, Lee S H, Cheong Y H, et al. 2001. A novel cold-inducible zinc finger protein from soybean, SCOF-1, enhances cold tolerance in transgenic plants. The Plant Journal, 25(3): 247-259.
Li Y, Su X, Zhang B, et al. 2009. Expression of jasmonic ethylene responsive factor gene in transgenic poplar tree leads to increased salt tolerance. Tree Physiology, 29(2): 273-279.
Mittler R, Kim Y, Song L, et al. 2006. Gain- and loss-of-function mutations in Zat10 enhance the tolerance of plants to abiotic stress. FEBS Letters, 580(28): 6537-6542.
Mukhopadhyay A, Vij S, Tyagi A K. 2004. Overexpression of a zinc-finger protein gene from rice confers tolerance to cold, dehydration, and salt stress in transgenic tobacco. Proceedings of the National Academy of Sciences of the United States of America, 101(16): 6309-6314.
Nanjo T, Kobayashi M, Yoshiba Y, et al. 1999. Biological functions of proline in morphogenesis and osmotolerance revealed in antisense transgenic Arabidopsis thaliana. Plant J, 18(2): 185-193.
Ravikumar R L, Patil B S, Salimath P M. 2003. Drought tolerance in sorghum by pollen selection using osmotic stress. Euphytica, 133(3): 371-376.
Sakamoto H, Araki T, Meshi T, et al. 2000. Expression of a subset of the Arabidopsis Cys2/His2-type zinc-finger protein gene family under water stress. Gene, 248(1): 23-32.
Sakamoto H, Maruyama K, Sakuma Y, et al. 2004. Arabidopsis Cys2/His2-type zinc-finger proteins function as transcription repressors under drought, cold, and high-salinity stress conditions. Plant Physiology, 136(1): 2734-2746.
Sinclair T R, Ludlow M M. 1986. Influence of soil water supply on the plant water balance of four tropical grain legumes. Australian Journal of Plant Physiology, 13(3): 329-341.
Sugano S, Kaminaka H, Rybka Z, et al. 2003. Stress-responsive zinc finger gene ZPT 2-3 plays a role in drought tolerance in petunia. The Plant Journal, 36(6): 830-841.
Takatsuji H. 1998. Zinc-finger transcription factors in plants. Cellular and Molecular Life Sciences CMLS, 54(6): 582-596.
Vogel J T, Zarka D G, Van Buskirk H A, et al. 2005. Roles of the CBF2 and ZAT12 transcription factors in configuring the low temperature transcriptome of Arabidopsis. The Plant Journal, 41(2): 195-211.
Xu S, Wang X, Chen J. 2007. Zinc finger protein 1 (ThZF1) from salt cress (Thellungiella halophila) is a Cys-2/His-2-type transcription factor involved in drought and salt stress. Plant Cell Reports, 26(4): 497-506.
Yamaguchi-Shinozaki K, Shinozaki K. 1994. A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low-temperature, or high-salt stress. The Plant Cell Online, 6(2): 251-264.
Yang X, Sun C, Hu Y, et al. 2008. Molecular cloning and characterization of a gene encoding RING zinc finger ankyrin protein from drought-tolerant Artemisia desertorum. Journal of Biosciences, 33(1): 103-112.
Zhu Z, Shi J, Xu W, et al. 2013. Three ERF transcription factors from Chinese wild grapevine Vitis pseudoreticulata participate in different biotic and abiotic stress-responsive pathways. J Plant Physiol, 170(10): 923-933.
Zou M, Guan Y, Ren H, et al. 2008. A bZIP transcription factor, OsABI5, is involved in rice fertility and stress tolerance. Plant Molecular Biology, 66(6): 675-683.

[1]	Lu Huijun, Li Ziyi, Liang Hanyu, Yue Yuanzhi, Zhou Tianchang, Yang Yuzhang, Wang Yucheng, Ji Xiaoyu. Expression and Stress Tolerance Analysis of NAC24 from Tamarix hispida [J]. Scientia Silvae Sinicae, 2019, 55(3): 54-63.
[2]	Zhang Jiangtao, Yang Shuhong, Zhu Di, Zhu Yanlin, Liu Youquan. Physiological Response of Annual Grafted Seedlings of Poplar 2025 and Its Two Bud Mutation Varieties to Drought Stress and Evaluation of Drought Resistance [J]. Scientia Silvae Sinicae, 2018, 54(6): 33-43.
[3]	Chen Wenrong, Pan Xia, Shao Junyi, Liao Fanglei, Yang Li, Hu Yingying, Yu Ying, Guo Weidong. Cloning,Expression and Analysis of Drought Resistance of VcLon1 Gene from Blueberry [J]. Scientia Silvae Sinicae, 2018, 54(6): 73-84.
[4]	Gu Zhanying, Yang Ruonan, Chen Hao. The Establishment of Isolation and Transient Transformation Methods of Protoplasts of Vernicia fordii Mesophyll Cells [J]. Scientia Silvae Sinicae, 2018, 54(1): 46-53.
[5]	Tang Feifei, Zhao Yulin, Wang Peilong, Feng Deming, Song Yi, Gao Caiqiu. Cloning and Stress Tolerance Analysis of ThP5CR from Tamarix hispida [J]. Scientia Silvae Sinicae, 2017, 53(7): 1-9.
[6]	Du Mingfeng, Ding Guijie, Zhao Xizhou. Responses to Continuous Drought Stress and Drought Resistance of Different Masson Pine Families [J]. Scientia Silvae Sinicae, 2017, 53(6): 21-29.
[7]	Luo Qinghong, Ning Husen, He Miao, Ji Xiaomin, Lei Chunying. Ecophysiological Responses of Five Sandy Shrubs to Drought Stress [J]. Scientia Silvae Sinicae, 2017, 53(11): 29-42.
[8]	Cheng Zhenxia, Hu Haitao, Yang Li, Wang Changchun, Guo Weidong, Yang Ling. Overexpression of EutPDS Gene from Elaeagnus umbellata Increases Lycopene Content in Tomato Fruit [J]. Scientia Silvae Sinicae, 2017, 53(1): 62-69.
[9]	Gao Runmei, Shi Xiaodong, Wang Lin, Han Na. Drought Resistance of One-Year-Old Seedlings of Larix principis-rupprechtii [J]. Scientia Silvae Sinicae, 2015, 51(7): 148-156.
[10]	Ye Songtao, Du Xuhua, Song Shuaijie, Li Li, Lu Yang, Ying Yeqing. Effect of Salicylic Acid on Physiological and Biochemical Characteristics of Phyllostachys edulis Seedlings under Drought Stress [J]. Scientia Silvae Sinicae, 2015, 51(11): 25-31.
[11]	Luo Zaiqi, Guo Huili, Yang Yadong, Yang Mingfeng, Ma Lanqin, Wang Younian. Agrobacterium-Mediated Transformation of Resveratrol Synthase Gene (PcPKS5) into Huping Jujube (Zizyphus jujuba) [J]. Scientia Silvae Sinicae, 2015, 51(10): 101-109.
[12]	Xing Cunwang, Huang Xuanrui, Li Yuling, Ma Zengwang, Fu Xiaoyan. Ecological Stability of Sand-Fixed Plantations in Huangyangtan [J]. Scientia Silvae Sinicae, 2014, 50(5): 101-108.
[13]	Li Simeng;Wang Yonglin;Huang Donghui;Tian Chengming. Establishment of a PEG-Mediated Genetic Transformation System and Expression of Green Fluorescence Protein in Colletotrichum gloeosporioides [J]. , 2013, 49(5): 121-127.
[14]	Ouyang Lejun;Liu Yuan;Huang Zhenchi;Zeng Fuhua. Transformation of TSRF 1 into Eucalyptus urophylla and the Broad-Spectrum Disease Resistance of the Transgenic Plant to Diseases [J]. Scientia Silvae Sinicae, 2013, 49(4): 46-53.
[15]	Li Xiaorui;Hu Shanglian;Cao Ying;Lu Xueqin;Ren Peng;Wu Xiaoyu;Zhou Meijuan. Agrobacterium-Mediated Transformation of 4CL Gene from Neosinocalamus affinis into Dendrocalamus farinosus [J]. Scientia Silvae Sinicae, 2012, 48(3): 38-44.

Transformation of Zinc Finger Protein Transcription Factor Gene (ZxZF) and Preliminary Evaluation of Drought Tolerance in Populus × euramericana

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments