5个毛果杨PtrZFP基因的鉴定和表达分析

doi:10.7525/j.issn.1673-5102.2020.02.011

摘要/Abstract

摘要： ZFP转录因子是植物中的一类具有指环结构域的转录因子。从毛果杨中鉴定出5条ZFP基因（命名为PtrZFP1-5），对其特性和表达模式进行了分析，以期初步了解这些基因是否能对胁迫做出应答。对PtrZFP1-5基因进行生物学分析，进一步利用qRT-PCR技术分析NaCl、PEG₆₀₀₀和ABA胁迫处理后毛果杨根、茎和叶中5条基因的表达情况。PtrZFP1-5基因编码蛋白氨基酸残基数为258~338 aa，编码蛋白的分子量为27.7~37.3 kDa，理论等电点为4.87~8.61，5个基因不均等的分布在毛果杨基因组的3条染色体上。qRT-PCR结果显示，0.2 mol·L^-1 NaCl、15%（w/v）PEG6000和100 μmol·L^-1 ABA胁迫处理后，5个PtrZFP基因在毛果杨根、茎和叶中的表达模式明显不同。PtrZFP1基因在3种胁迫后毛果杨中均被明显的上调表达；PtrZFP2基因在盐、渗透和ABA胁迫处理后，叶中的表达都明显被抑制；PtrZFP3基因受到干旱胁迫时在根中的响应最为明显；而叶和茎中，表达量在大部分胁迫的大部分时间点无明显改变。PtrZFP4基因也能在根和茎中对干旱胁迫做出明显应答。PtrZFP5基因在经受盐和ABA胁迫后，在叶中的表达受到明显抑制。PtrZFP1-5这5个基因至少能在一种器官中对一种胁迫处理做出应答，但参与的胁迫应答类型和机制可能不同。

关键词: 锌指蛋白, 转录因子, 毛果杨, 非生物胁迫

Abstract: ZFPs are a class of transcription factors in plants that have a loop domain. In this study, 5 ZFP genes(named PtrZFP1-5) were identified from Populus trichocarpa, and their sequence characteristics and expression patterns were analyzed to understand the stress response function. The bioinformatics analysis of PtrZFP1-5 gene was carried out, and the expressions of 5 PtrZFP genes in root, stem and leaf of Populus under NaCl, PEG₆₀₀₀ and ABA treatment were further analyzed by qRT-PCR. The number of amino acid residues encoded by PtrZFP1-5 gene was 258-338 aa, the molecular weight of the encoded protein was 27.7-37.3 kDa,and the theoretical isoelectric point was 4.87-8.61. 5 PtrZFP genes were unevenly distributed on the three chromosomes of Populus genome. The results of qRT-PCR showed that the expression patterns of 5 PtrZFP genes in roots, stems and leaves of Populus were significantly different after treatment with 0.2M NaCl, 15%(w/v) PEG6000or 100μM ABA treatment. The expressions of PtrZFP1 were all up-regulated in three kinds of stresses. The expressions of PtrZFP2 were significantly inhibited in leaves after salt, osmotic and ABA treatment. The expression changed of PtrZFP3 in roots was most obvious under drought stress. In leaves and stems, the expression level did not change significantly at most of the time points of most stress. The PtrZFP4 gene also responded to drought stress in roots and stems. The expression of PtrZFP5 gene in leaves was significantly reduced after exposure to salt and ABA stress. PtrZFP1-5 genes can respond to at least one stress treatment in one organ, but the type and mechanism of the involved stress response may be different.

Key words: ZFP, transcription factor, Populus trichocarpa, abiotic stress

中图分类号:

Q943.2

李亚博, 吕佳欣, 谭冰, 高彩球. 5个毛果杨PtrZFP基因的鉴定和表达分析[J]. 植物研究, 2020, 40(2): 243-250.

LI Ya-Bo, Lü Jia-Xin, TAN Bing, GAO Cai-Qiu. Cloning and expression analysis of 5 ZFP genes from Poplus trichocarpa[J]. Bulletin of Botanical Research, 2020, 40(2): 243-250.

参考文献 22

[1]	黄骥,王建飞,张红生.植物C2H2型锌指蛋白的结构与功能[J].遗传,2004,26(3):414-418.Huang J,Wang J F,Zhang H S.Structure and function of plant C2H2 zinc finger protein[J].Hereditas,2004,26(3):414-418.
[2]	Chai G H,Hu R B,Zhang D Y,et al.Comprehensive analysis of CCCH zinc finger family in poplar(Populus trichocarpa)[J].BMCGenomics,2012,13:253,doi:10.1186/1471-2164-13-253.
[3]	Gupta S K,Rai A K,Kanwar S S,et al.Comparative analysis of zinc finger proteins involved in plant disease resistance[J].PLoS One,2012,7(8):e42578,doi:10.1371/journal.pone.0042578.
[4]	Jan A,Maruyama K,Todaka D,et al.OsTZF1,a CCCH-Tandem Zinc Finger protein,confers delayed senescence and stress tolerance in rice by regulating stress-related Genes[J].Plant Physiology,2013,161(3):1202-1216,doi:10.1104/pp.112.205385.
[5]	Mak,Maoj H,Lix H,et al.Sequence and expression analysis of the C3HC4-type RINGfinger gene family in rice[J].Gene,2009,444(1-2):33-45,doi:10.1016/j.gene.2009.05.018.
[6]	Gao Y S,Nishikawa H,Badejo A A,et al.Expression of aspartyl protease and C3HC4-type RING zinc finger genes are responsive to ascorbic acid in Arabidopsis thaliana[J].Journal of Experimental Botany,2011,62(10):3647-3657,doi:10.1093/jxb/err068.
[7]	Zeba N,Isbat M,Kwon N J,et al.Heat-inducible C3HC4 type RING zinc finger protein gene from Capsicum annuum enhances growth of transgenic tobacco[J].Planta,2009,229(4):861-871,doi:10.1007/s00425-008-0884-0.
[8]	Hang L,Wang M J,Yuan F,et al.The CCCH zinc finger protein gene AtZFP1 improves salt resistance in Arabidopsis thaliana[J].Plant Molecular Biology,2014,86(3):237-253,doi:10.1007/s11103-014-0226-5.
[9]	Zhang H,Liu Y P,Wen F,et al.A novel rice C2H2-type zinc finger protein,ZFP36,is a key player involved in abscisic acid-induced antioxidant defence and oxidative stress tolerance in rice[J].Journal of Experimental Botany,2014,65(20):5795-5809,doi:10.1093/jxb/eru313.
[10]	An L J,Zhou Z J,Sun L L,et al.A zinc finger protein gene ZFP5 integrates phytohormone signaling to control root hair development in Arabidopsis[J].The Plant Journal,2012,72(3):474-490,doi:10.1111/j.1365-313X.2012.05094.x.
[11]	孔瑾,曹宛虹,张劲松,等.水稻受盐抑制基因OsZFP1的转基因分析[J].植物学报,2004,46(5):573-577.Kong J,Cao W H,Zhang J S,et al.Transgenic analysis of a salt-inhibited OsZFP1 gene from rice[J].Acta Botanica Sinica,2004,46(5):573-577.
[12]	Dathan N,Zaccaro L,Esposito S.The Arabidopsis SUPERMAN protein is able to specifically bind DNA through its single Cys2-His2 zinc finger motif[J].Nucleic Acids Research,2002,30(22):4945-4951,doi:10.1093/nar/gkf613.
[13]	Takatsuji H.Zinc-finger transcription factors in plants[J].Cellular and Molecular Life SciencesCMLS,1998,54(6):582-596,doi:10.1007/s000180050186.
[14]	Kobayashi A,Sakamoto A,Kubo K,et al.Seven zinc-finger transcription factors are expressed sequentially during the development of anthers in petunia[J].The Plant Journal,1998,13(4):571-576,doi:10.1046/j.1365-313X.1998.00043.x.
[15]	Lippuner V,Cyert M S,Gasser C S.Two classes of plant cDNA clones differentially complement yeast calcineurin mutants and increase salt tolerance of wild-type yeast[J].Journal of Biological Chemistry,1996,271(22):12859-12866,doi:10.1074/jbc.271.22.12859.
[16]	Sakamoto H,Araki T,Meshi T,et al.Expression of a subset of the Arabidopsis Cys2/His2-type zinc-finger protein gene family under water stress[J].Gene,2000,248(1-2):23-32,doi:10.1016/S0378-1119(00)00133-5.
[17]	秦智慧,杨青川,晁跃辉,等.CCCH型锌指蛋白研究进展[J].生物技术通报,2010(8):1-6.Qin Z H,Yang Q C,Chao Y H,et al.Study of CCCH-type zinc finger protein[J].Biotechnology Bulletin,2010(8):1-6.
[18]	Sun J Q,Jiang H L,Xu Y X,et al.The CCCH-Type zinc finger proteins AtSZF1 and AtSZF2 regulate salt stress responses in Arabidopsis[J].Plant and Cell Physiology,2007,48(8):1148-1158,doi:10.1093/pcp/pcm088.
[19]	Li Z S,Thomas T L.PEI1,an embryo-specific zinc finger protein gene required for heart-stage embryo formation in Arabidopsis[J].The Plant Cell,1998,10(3):383-398,doi:10.1105/tpc.10.3.383.
[20]	Grabowska A,Wisniewska A,Tagashira N,et al.Characterization of CsSEF1 gene encoding putative CCCH-type zinc finger protein expressed during cucumber somatic embryogenesis[J].Journal of Plant Physiology,2009,166(3):310-323,doi:10.1016/j.jplph.2008.06.005.
[21]	Kim D H,Yamaguchi S,Lim S,et al.SOMNUS,a CCCH-Type zinc finger protein in Arabidopsis,negatively regulates light-dependent seed germination downstream of PIL5[J].The Plant Cell,2008,20(5):1260-1277,doi:10.1105/tpc.108.058859.
[22]	Wang L,Xu Y Y,Zhang C,et al.OsLIC,a novel CCCH-Type zinc finger protein with transcription activation,mediates rice architecture via brassinosteroids signaling[J].PLoS One,2008,3(10):e3521,doi:10.1371/journal.pone.0003521.