• Scopus
  • Chinese Science Citation Database (CSCD)
  • A Guide to the Core Journal of China
  • CSTPCD
  • F5000 Frontrunner
  • RCCSE
Advanced search
ZHANG Xiao-fei, LU Xin, DUAN Hui, LIAN Cong-long, XIA Xin-li, YIN Wei-lun. Cloning and functional analysis of PeNAC045 from Populus euphratica[J]. Journal of Beijing Forestry University, 2015, 37(6): 1-10. DOI: 10.13332/j.1000-1522.20150066
Citation: ZHANG Xiao-fei, LU Xin, DUAN Hui, LIAN Cong-long, XIA Xin-li, YIN Wei-lun. Cloning and functional analysis of PeNAC045 from Populus euphratica[J]. Journal of Beijing Forestry University, 2015, 37(6): 1-10. DOI: 10.13332/j.1000-1522.20150066

Cloning and functional analysis of PeNAC045 from Populus euphratica

More Information
  • Received Date: March 15, 2015
  • NAC (NAM, ATAF1/2 and CUC2) domain proteins constitute one of the largest plant-specific transcription factors (TFs) and play an important role in regulating senescence, cell division, wood formation and biotic and abiotic stresses. In present study, we successfully isolated a stress responsive gene from Populus euphratica, i.e., PeNAC045. Sequencing results indicated that the length of PeNAC045 is 915 bp encoding 304 amino acids, and PeNAC045 shares 96.05% homology in amino acid sequence with PtrNAC045. The expression of PeNAC045 in response to NaCl and drought stress was characterized. PeNAC045 mRNA expression was strongly induced by high-salinity and drought treatment. The expression vector pBI121-PeNAC045-GFP was constructed using the full-length PeNAC045 cDNA cloned into the pBI121-GFP vector. After sequencing confirmation, the construct and positive control (empty vector) were transformed into Arabidopsis. Subcellular localization experiments in Arabidopsis indicated that the PeNAC045-GFP fusion protein was localized in the nucleus. Arabidopsis was stained with the DNA dye 4,6-diamidino-2-phenylindole (DAPI) to visualize the nucleus. The expression vector pCAMBIA1301-PeNAC045 was constructed and transformed into Arabidopsis thaliana wild type (Col-0) and ataf2 mutant using floral dip method. Then, we obtained the PeNAC045 overexpression lines of Arabidopsis and ataf2/PeNAC045. To test the function of PeNAC045, each line was treated with NaCl. The germination percentage of PeNAC045 overexpression lines of Arabidopsis was reduced and the root length was shorter under 150 mmol/L NaCl stress, compared to ataf2 mutant and wild-type plants. Furthermore, the sensitivity of transgenic PeNAC045 overexpression lines to NaCl stress was increased at the seedling stage and the height of the seedlings was significantly lower than others. Our results indicated that PeNAC045, as a transcriptional regulator, negatively regulates the expression of stress responsive genes under NaCl stress.
  • [1]
    NURUZZAMAN M, SHARONI A M, KIKUCHI S. Roles of NAC transcription factors in the regulation of biotic and abiotic stress responses in plants[J]. Frontiers in Microbiology, 2013, 4: 248.
    [1]
    MA H S, XIA X L, YIN W L. Constructing cDNA-AFLP reaction system of abiotic stress study for Populus euphratica[J]. Journal of Beijing Forestry University, 2010, 32(5): 34-40.
    [2]
    ERNST H A, OLSEN A N, SKRIVER K, et al. Structure of the conserved domain of ANAC, a member of the NAC family of transcription factors[J]. Embo Reports, 2004, 5(3): 297-303.
    [2]
    DUAN Z X, QIN Y R, XIA X L, et al. Overexpression of Populus euphratica peu-MIR156j gene enhancing salt tolerance in Arabidopsis thaliana[J]. Journal of Beijing Forestry University, 2012, 33(6): 1-7.
    [3]
    DUVAL M, HSIEH T F, KIM S Y, et al. Molecular characterization of AtNAM: a member of the Arabidopsis NAC domain superfamily[J]. Plant Molecular Biology, 2002, 50(2): 237-248.
    [3]
    MA H S, XIA X L, YIN W L. Cloning and analysis of SCL7 gene from Populus euphratica[J]. Journal of Beijing Forestry University, 2011, 33(1): 1-10.
    [4]
    AIDA M, ISHIDA T, FUKAKI H, et al. Genes involved in organ separation in Arabidopsis: an analysis of the cup-shaped cotyledon mutant[J]. Plant Cell, 1997, 9(6): 841-857.
    [4]
    QIN Y R, XIA X L, YIN W L. Expression determination of miR169g under dehydration and high salinity stress in Populus euphratica leaves by real-time quantitative PCR[J]. Modern Instruments, 2011, 17(3): 28-30.
    [5]
    OLSEN A N, ERNST H A, LO LEGGIO L, et al. NAC transcription factors: structurally distinct, functionally diverse[J]. Trends in Plant Science, 2005, 10(2): 79-87.
    [6]
    SU H Y, ZHANG S Z, YUAN X W, et al. Genome-wide analysis and identification of stress-responsive genes of the NAM-ATAF1,2-CUC2 transcription factor family in apple[J]. Plant Physiology and Biochemistry, 2013, 71: 11-21.
    [7]
    WANG Z Y, RASHOTTE A M, MOSS A G, et al. Two NAC transcription factors from Citrullus colocynthis, CcNAC1, CcNAC2 implicated in multiple stress responses[J]. Acta Physiologiae Plantarum, 2014, 36(3): 621-634.
    [8]
    LIU G Z, LI X L, JIN S X, et al. Overexpression of rice NAC gene SNAC1 improves drought and salt tolerance by enhancing root development and reducing transpiration rate in transgenic cotton[J]. PLoS One, 2014, 9(1): e86895.
    [9]
    LI W, HUANG G Q, ZHOU W, et al. A cotton (Gossypium hirsutum) gene encoding a NAC transcription factor is involved in negative regulation of plant xylem development[J]. Plant Physiology and Biochemistry, 2014, 83: 134-141.
    [10]
    FAN K, WANG M, MIAO Y, et al. Molecular evolution and expansion analysis of the NAC transcription factor in Zea mays[J]. PLoS One, 2014, 9(11): e111837.
    [11]
    ZHOU Y, HUANG W F, LIU L, et al. Identification and functional characterization of a rice NAC gene involved in the regulation of leaf senescence[J]. BMC Plant Biology, 2013, 13(1): 132.
    [12]
    WANG J Y, WANG J P, HE Y. A Populus euphratica NAC protein regulating Na+/K+ homeostasis improves salt tolerance in Arabidopsis thaliana[J]. Gene, 2013, 521(2): 265-273.
    [13]
    RIECHMANN J L, HEARD J, MARTIN G, et al. Arabidopsis transcription factors: genome-wide comparative analysis among eukaryotes[J]. Science, 2000, 290: 2105-2110.
    [14]
    NURUZZAMAN M, MANIMEKALAI R, SHARONI A M, et al. Genome-wide analysis of NAC transcription factor family in rice[J]. Gene, 2010, 465(1-2): 30-44.
    [15]
    HU R B, QI G A, KONG Y Z, et al. Comprehensive analysis of NAC domain transcription factor gene family in Populus trichocarpa[J]. Bmc Plant Biology, 2010, 10(1): 145.
    [16]
    TRAN L S P, NAKASHIMA K, SAKUMA Y, et al. Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 promoter[J]. Plant Cell, 2004, 16(9): 2481-2498.
    [17]
    LU P L, CHEN N Z, AN R, et al. A novel drought-inducible gene, ATAF1, encodes a NAC family protein that negatively regulates the expression of stress-responsive genes in Arabidopsis[J]. Plant Molecular Biology, 2007, 63(2): 289-305.
    [18]
    JENSEN M K, LINDEMOSE S, DE MASI F, et al. ATAF1 transcription factor directly regulates abscisic acid biosynthetic gene NCED3 in Arabidopsis thaliana[J]. Febs Open Bio, 2013, 3: 321-327.
    [19]
    OH S K, LEE S, YU S, et al. Expression of a novel NAC domain-containing transcription factor (CaNAC1) is preferentially associated with incompatible interactions between chili pepper and pathogens[J]. Planta, 2005, 222(5): 876-887.
    [20]
    DELESSERT C, WILSON I W, VAN DER STRAETEN D, et al. Spatial and temporal analysis of the local response to wounding in Arabidopsis leaves[J]. Plant Molecular Biology, 2004, 55(2): 165-181.
    [21]
    DELESSERT C, KAZAN K, WILSON I W, et al. The transcription factor ATAF2 represses the expression of pathogenesis-related genes in Arabidopsis[J]. Plant Journal, 2005, 43(5): 745-757.
    [22]
    WANG X, CULVER J N. DNA binding specificity of ATAF2, a NAC domain transcription factor targeted for degradation by tobacco mosaic virus[J]. Bmc Plant Biology, 2012, 12(1): 157.
    [23]
    HUH S U, LEE S B, KIM H H, et al. ATAF2, a NAC transcription factor, binds to the promoter and regulates NIT2 gene expression involved in auxin biosynthesis[J]. Molecules and Cells, 2012, 34(3): 305-313.
    [24]
    ZHONG R Q, LEE C H, YE Z H. Functional characterization of poplar wood-associated NAC domain transcription factors[J]. Plant Physiology, 2010, 152(2): 1044-1055.
    [25]
    ZHAO Y J, SUN J Y, XU P, et al. Intron-mediated alternative splicing of WOOD-ASSOCIATED NAC TRANSCRIPTION FACTOR1B regulates cell wall thickening during fiber development in Populus species[J]. Plant Physiology, 2014, 164(2): 765-776.
    [26]
    GU R S, LIU Q L, PEI D, et al. Understanding saline and osmotic tolerance of Populus euphratica suspended cells[J]. Plant Cell Tissue and Organ Culture, 2004, 78(3): 261-265.
    [27]
    LI B S, YIN W L, XIA X L. Identification of microRNAs and their targets from Populus euphratica[J]. Biochemical and Biophysical Research Communications, 2009, 388(2): 272-277.
    [28]
    马洪双, 夏新莉, 尹伟伦. 建立胡杨抗逆研究的cDNA-AFLP反应体系[J]. 北京林业大学学报, 2010, 32(5): 34-40.
    [29]
    段中鑫, 覃玉蓉, 夏新莉, 等. 超量表达胡杨peu-MIR156j 基因增强拟南芥耐盐性[J]. 北京林业大学学报, 2012, 33(6): 1-7.
    [30]
    马洪双, 夏新莉, 尹伟伦. 胡杨SCL7基因及其启动子片段的克隆与分析[J]. 北京林业大学报, 2011, 33(1): 1-10.
    [31]
    覃玉蓉, 夏新莉, 尹伟伦. 实时荧光定量PCR检测miR169g在脱水与高盐胁迫下胡杨叶中的表达[J]. 现代仪器, 2011, 17(3): 28-30.
    [32]
    OHTANI M, NISHIKUBO N, XU B, et al. A NAC domain protein family contributing to the regulation of wood formation in poplar[J]. Plant Journal, 2011, 67(3): 499-512.
    [33]
    GOODSTEIN D M, SHU S Q, HOWSON R, et al. Phytozome: a comparative platform for green plant genomics[J]. Nucleic Acids Research, 2012, 40(D1): 1178-1186.
    [34]
    SAITOU N, NEI M. The neighbor-joining method: a new method for reconstructing phylogenetic trees[J]. Molecular Biology and Evolution, 1987, 4(4): 406-425.
    [35]
    WANG H L, CHEN J H, TIAN Q Q, et al. Identification and validation of reference genes for Populus euphratica gene expression analysis during abiotic stresses by quantitative real-time PCR[J]. Physiologia Plantarum, 2014, 152(3): 529-545.
    [36]
    ZHANG X R, HENRIQUES R, LIN S S, et al. Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method[J]. Nature Protocols, 2006, 1(2): 641-646.
    [37]
    JEONG J S, KIM Y S, BAEK K H, et al. Root-specific expression of OsNAC10 improves drought tolerance and grain yield in rice under field drought conditions[J]. Plant Physiology, 2010, 153(1): 185-197.
    [38]
    SONG S Y, CHEN Y, CHEN J, et al. Physiological mechanisms underlying OsNAC5-dependent tolerance of rice plants to abiotic stress[J]. Planta, 2011, 234(2): 331-345.
    [39]
    LU M, YING S, ZHANG D F, et al. A maize stress-responsive NAC transcription factor, ZmSNAC1, confers enhanced tolerance to dehydration in transgenic Arabidopsis[J]. Plant Cell Reports, 2012, 31(9): 1701-1711.
    [40]
    HU H H, YOU J, FANG Y J, et al. Characterization of transcription factor gene SNAC2 conferring cold and salt tolerance in rice[J]. Plant Molecular Biology, 2008, 67(1-2): 169-181.
    [41]
    CHEN S L, LI J K, WANG S S, et al. Effects of NaCl on shoot growth, transpiration, ion compartmentation, and transport in regenerated plants of Populus euphratica and Populus tomentosa[J]. Canadian Journal of Forest Research, 2003, 33(6): 967-975.
    [42]
    OTTOW E A, BRINKER M, TEICHMANN T, et al. Populus euphratica displays apoplastic sodium accumulation, osmotic adjustment by decreases in calcium and soluble carbohydrates, and develops leaf succulence under salt stress[J]. Plant Physiology, 2005, 139(4): 1762-1772.
    [43]
    HAN X, TANG S, AN Y, et al. Overexpression of the poplar NF-YB7 transcription factor confers drought tolerance and improves water-use efficiency in Arabidopsis[J]. Journal of Experimental Botany, 2013, 64(14): 4589-4601.
  • Cited by

    Periodical cited type(5)

    1. 乔志宏,侯宏宇,高梅香,卢廷玉. 短时暴雨对小兴安岭凉水阔叶红松林地表甲虫群落的影响. 生态学报. 2020(14): 4994-5007 .
    2. 郑欣颖,佘汉基,薛立,蔡金桓. 外源性氮和磷对火力楠凋落叶分解的影响. 华南农业大学学报. 2018(01): 98-104 .
    3. 李旭华,孙建新. Biome-BGC模型模拟阔叶红松林碳水通量的参数敏感性检验和不确定性分析. 植物生态学报. 2018(12): 1131-1144 .
    4. 毛宏蕊,金光泽. 氮添加对典型阔叶红松林净初级生产力的影响. 北京林业大学学报. 2017(08): 42-49 . 本站查看
    5. 宋蕾,林尤伟,金光泽. 模拟氮沉降对典型阔叶红松林土壤微生物群落特征的影响. 南京林业大学学报(自然科学版). 2017(05): 7-12 .

    Other cited types(9)

Catalog

    Article views (2285) PDF downloads (46) Cited by(14)

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return