Cloning and expression characteristics of PdMODD genes in Populus × euramericana

doi:10.12302/j.issn.1000-2006.202007015

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2021, Vol. 45 ›› Issue (2): 43-50.doi: 10.12302/j.issn.1000-2006.202007015

Previous Articles Next Articles

Cloning and expression characteristics of PdMODD genes in Populus × euramericana

ZHANG Tengqian¹(), ZHANG Weixi¹, DING Changjun¹, ZHANG Jing¹, HU Zanmin², SU Xiaohua¹^,^*()

1. State Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of National Forestry and Grassland Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China
2. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China

Received:2020-07-15 Accepted:2020-11-30 Online:2021-03-30 Published:2021-04-09
Contact: SU Xiaohua E-mail:tengqianzhang66@126.com;suxh@caf.ac.cn

Abstract

Abstract:

【Objective】To explore the function of PdMODD in the growth and development of Populus × euramericana, we analyzed their sequence characteristics, subcellular localization and expression patterns in response to salt and drought stress, which will provide a theoretical basis for revealing the molecular mechanisms of stress resistance and breeding new poplar varieties. 【Method】Based on the rice MODD amino acid sequence, Podel.08G114100, Podel.10G158900 and Podel.17G098500 were selected using BLAST in the genome of Populus deltoides WV94 v2.1. Using these gene sequences as references and Populus × euramericana ‘Bofeng 3 hao’ cDNA as template, three genes (PdMODD1,PdMODD2 and PdMODD3) were cloned. The structural characteristics of the PdMODD protein and its genetic relationship with its homologous proteins were analyzed using bioinformatics methods. The subcellular localization of PdMODD genes was analyzed using particle bombardment. The expression characteristics of PdMODD genes in different tissues and their response to different stresses were analyzed using qRT-PCR. 【Results】The open reading frames of PDMODD1-PDMODD3 genes were 1 065, 1 065 and 1 074 bp, encoding 354, 354 and 357 amino acids, respectively, all of which were unstable hydrophilic proteins located in the nucleus.PdMODD genes are all members of the NINJA family and contain three conserved domains, one of which is the transcriptional repression motif EAR. Phylogenetic analysis demonstrated that PdMODD1 protein was closely related to PtAFP2 (Populus trichocarpa) and located in the same branch as AtAFP1 (Arabidopsis thaliana) and MeAFP2 (Manihot esculenta). PdMODD2 protein had a close relationship with JcAFP2 (Jatropha curcas) and converged into a branch with AtAFP2; PdMODD3 was closely related to PtAFP3-1 and located in the same branch as QsAFP3 (Quercus suber) and AtAFP4. The subcellular localization results showed that PdMODD genes were localized in the nucleus. The results of tissue specificity analysis showed that PdMODD genes were highly expressed in the leaves and the least expressed in the roots. The expression patterns of stress response showed that under NaCl and PEG stress, PdMODD1, PdMODD2 and PdMODD3 were significantly upregulated in the root, stem and leaf tissues. 【Conclusion】PdMODD1-PdMODD3 are all members of the NINJA family, with the conserved transcriptional repression motif EAR and tissue specificity. They were highly expressed in the leaves, significantly induced by NaCl and PEG stress, and were in a branch with AtAFP1, AtAFP2 and AtAFP4, which had negative regulatory effects on ABA and salt stress. It is hypothesized that PdMODD1, PdMODD2 and PdMODD3 may play an important regulatory role in response to P. euramericana ‘Bofeng 3 hao’ salt and drought stress.

Key words: Populus × euramaricana ‘Bofeng 3 hao’, PdMODD, bioinformatics analysis, expression pattern analysis

CLC Number:

S785.5

ZHANG Tengqian, ZHANG Weixi, DING Changjun, ZHANG Jing, HU Zanmin, SU Xiaohua. Cloning and expression characteristics of PdMODD genes in Populus × euramericana[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2021, 45(2): 43-50.

Figures/Tables 8

Table 1

Fig.1

Table 2

Fig.2

Fig.3

Fig.4

Fig.5

Fig.6

References 30

[1]	KAGALE S, ROZWADOWSKI K. EAR motif-mediated transcriptional repression in plants[J]. Epigenetics, 2011,6(2):141-146.DOI: 10.4161/epi.6.2.13627.
[2]	张健飞, 权瑞党, 黄荣峰. EAR转录抑制子结构及功能的研究[J]. 中国农业科技导报, 2011,13(4):53-57.
	ZHANG J F, QUAN R D, HUANG R F. Studies on structure and function of repressors with EAR motif[J]. J Agric Sci Technol, 2011,13(4):53-57.DOI: 10.3969/j.issn.1008-0864.2011.04.08.
[3]	TANG N, MA S, ZONG W, et al. MODD mediates deactivation and degradation of OsbZIP46 to negatively regulate ABA signaling and drought resistance in rice[J]. Plant Cell, 2016,28(9):2161-2177.DOI: 10.1105/tpc.16.00171.
[4]	GARCIA M E, LYNCH T, PEETERS J, et al. A small plant-specific protein family of ABI five binding proteins (AFPs) regulates stress response in germinating Arabidopsis seeds and seedlings[J]. Plant Mol Biol, 2008,67(6):643-658.DOI: 10.1007/s11103-008-9344-2.
[5]	PAUWELS L, BARBERO G F, GEERINCK J, et al. NINJA connects the co-repressor TOPLESS to jasmonate signalling[J]. Nature, 2010,464(7289):788-791.DOI: 10.1038/nature08854.
[6]	KAZAN K, MANNERS J M. JAZ repressors and the orchestration of phytohormone crosstalk[J]. Trends Plant Sci, 2012,17(1):22-31.DOI: 10.1016/j.tplants.2011.10.006.
[7]	DE GEYTER N, GHOLAMI A, GOORMACHTIG S, et al. Transcriptional machineries in jasmonate-elicited plant secondary metabolism[J]. Trends Plant Sci, 2012,17(6):349-359.DOI: 10.1016/j.tplants.2012.03.001.
[8]	MA S, TANG N, LI X, et al. Reversible histone H2B monoubiquitination fine-tunes abscisic acid signaling and drought response in rice[J]. Mol Plant, 2019,12(2):263-277.DOI: 10.1016/j.molp.2018.12.005.
[9]	XIANG Y, TANG N, DU H, et al. Characterization of OsbZIP23 as a key player of the basic leucine zipper transcription factor family for conferring abscisic acid sensitivity and salinity and drought tolerance in rice[J]. Plant Physiol, 2008,148(4):1938-1952.DOI: 10.1104/pp.108.128199.
[10]	LU G J, GAO C X, ZHENG X N, et al. Identification of OsbZIP72 as a positive regulator of ABA response and drought tolerance in rice[J]. Planta, 2009,229(3):605-615.DOI: 10.1007/s00425-008-0857-3.
[11]	王倩姿, 王玉, 孙志梅, 等. 腐植酸类物质的施用对盐碱地的改良效果[J]. 应用生态学报, 2019,30(4):1227-1234.
	WANG Q Z, WANG Y, SUN Z M, et al. Amelioration effect of humic acid on saline-alkali soil[J]. Chin J Appl Ecol, 2019,30(4):1227-1234.DOI: 10.13287/j.1001-9332.201904.001.
[12]	徐淑平, 卫志明. 基因枪的使用方法介绍[J]. 植物生理学通讯, 1998,34(1):41-43.
	XU S P, WEI Z M. Introduction to method of microprojectile bombardment and its application[J]. Plant Physiol Commun, 1998,34(1):41-43.DOI: 10.13592/j.cnki.ppj.1998.01.014.
[13]	LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method[J]. Methods (San Diego Calif), 2001,25(4):402-408.DOI: 10.1006/meth.2001.1262.
[14]	刘中原, 姜波, 吕佳欣, 等. 刚毛柽柳Th2CysPr_x基因的互作蛋白及其表达模式分析[J]. 南京林业大学学报(自然科学版), 2019,43(2):86-92.
	LIU Z Y, JIANG B, LYU J X, et al. Interacting proteins of Tamarix hispida Th2CysPr_x and their expression pattern analysis[J]. J Nanjing For Univ (Nat Sci Ed), 2019,43(2):86-92.DOI: 10.3969/j.issn.1000-2006.201806018.
[15]	杜娟, 柴友荣. 植物转录抑制子的结构特征及其作用机理[J]. 植物学通报, 2008,25(3):344-353.
	DU J, CHAI Y R. Structural features and action mechanisms of plant transcriptional repressors[J]. Chin Bull Bot, 2008,25(3):344-353.DOI: 10.3969/j.issn.1674-3466.2008.03.012.
[16]	KAGALE S, LINKS M G, ROZWADOWSKI K. Genome-wide analysis of ethylene-responsive element binding factor-associated amphiphilic repression motif-containing transcriptional regulators in Arabidopsis[J]. Plant Physiol, 2010,152(3):1109-1134.DOI: 10.1104/pp.109.151704.
[17]	WEIGEL R R, PFITZNER U M, GATZ C. Interaction of NIMIN₁ with NPR1 modulates PR gene expression in Arabidopsis[J]. Plant Cell, 2005,17(4):1279-1291.DOI: 10.1105/tpc.104.027441.
[18]	CHERN M, CANLAS P E, FITZGERALD H A, et al. Rice NRR,a negative regulator of disease resistance,interacts with Arabidopsis NPR1 and rice NH₁[J]. Plant J, 2005,43(5):623-635.DOI: 10.1111/j.1365-313x.2005.02485.x.
[19]	OHTA M, MATSUI K, HIRATSU K, et al. Repression domains of class II ERF transcriptional repressors share an essential motif for active repression[J]. Plant Cell, 2001,13(8):1959-1968.DOI: 10.1105/tpc.010127.
[20]	LIU W, KAREMERA N J U, WU T, et al. The ethylene response factor AtERF₄ negatively regulates the iron deficiency response in Arabidopsis thaliana[J]. PLoS One, 2017,12(10):e0186580.DOI: 10.1371/journal.pone.0186580.
[21]	YAISH M W, EL-KEREAMY A, ZHU T, et al. The APETALA-2-like transcription factor OsAP2-39 controls key interactions between abscisic acid and gibberellin in rice[J]. PLoS Genet, 2010,6(9):e1001098.DOI: 10.1371/journal.pgen.1001098.
[22]	WAN L, ZHANG J, ZHANG H, et al. Transcriptional activation of OsDERF₁ in OsERF₃ and OsAP2-39 negatively modulates ethylene synjournal and drought tolerance in rice[J]. PLoS One, 2011,6(9):e25216.DOI: 10.1371/journal.pone.0025216.
[23]	ZHANG H W, ZHANG J F, QUAN R D, et al. EAR motif mutation of rice OsERF₃ alters the regulation of ethylene biosynjournal and drought tolerance[J]. Planta, 2013,237(6):1443-1451.DOI: 10.1007/s00425-013-1852-x.
[24]	PAN I C, LI C W, SU R C, et al. Ectopic expression of an EAR motif deletion mutant of SlERF₃ enhances tolerance to salt stress and Ralstonia solanacearum in tomato[J]. Planta, 2010,232(5):1075-1086.DOI: 10.1007/s00425-010-1235-5.
[25]	HUANG J, YANG X, WANG M M, et al. A novel rice C₂H₂-type zinc finger protein lacking DLN-box/EAR-motif plays a role in salt tolerance[J]. Biochim et Biophys Acta (BBA)-Gene Struct Expr, 2007,1769(4):220-227.DOI: 10.1016/j.bbaexp.2007.02.006.
[26]	LOPEZ-MOLINA L, MONGRAND S, KINOSHITA N, et al. AFP is a novel negative regulator of ABA signaling that promotes ABI₅ protein degradation[J]. Genes Dev, 2003,17(3):410-418.DOI: 10.1101/gad.1055803.
[27]	OHNISHI N, HIMI E, YAMASAKI Y, et al. Differential expression of three ABA-insensitive five binding protein (AFP)-like genes in wheat[J]. Genes Genet Syst, 2008,83(2):167-177.DOI: 10.1266/ggs.83.167.
[28]	WU J, SENG S, CARIANOPOL C, et al. Cloning and characterization of a novel Gladiolus hybridus AFP family gene (GhAFP-like) related to corm dormancy[J]. Biochem Biophys Res Commun, 2016,471(1):198-204.DOI: 10.1016/j.bbrc.2016.01.146.
[29]	HUANG M D, WU W L. Overexpression of TMAC2,a novel negative regulator of abscisic acid and salinity responses,has pleiotropic effects in Arabidopsis thaliana[J]. Plant Mol Biol, 2007,63(4):557-569.DOI: 10.1007/s11103-006-9109-8.
[30]	CAO M J, ZHANG Y L, LIU X, et al. Combining chemical and genetic approaches to increase drought resistance in plants[J]. Nat Commun, 2017,8(1):1183.DOI: 10.1038/s41467-017-01239-3.

Metrics

Comments

www.nldxb.njfu.edu.cn

Edited ＆ Published by Editorial Department of Nanjing Forestry University(Natural Sciences Edition)
Address： No.159 Longpan Road,Nanjing 210037 Jiangsu,P.R.China
E-mail ：xuebao@njfu.edu.cn , xuebao@njfu.com.cn
Telephone +86-25-85428247,85427076
Distributed by China International Book Trading Corporation P.O. Box 399, Beijing ,P.R. China

用途 application	引物名称 primer name	上游引物序列(5'-3') forward primer sequence	下游引物序列(5'-3') reverse primer sequence
PdMODD基因克隆 cloning of PdMODD genes	PdMODD1	TAGAGAGGCAGCATCATGGGAG	TTACACAAAGGACGGACCAG
	PdMODD2	ATGGGAGAAGCAAACGCGAAT	TTATTCAAAGGACGGACAAGAAGG
	PdMODD3	GCTGGGATTTGAGGTTGTTGGT	TTAAAAAAAGGGAGAGGGGTGAACC
qRT-PCR	PdMODD1-RT	ATCAAGCAGTGGTGGTGAAG	CCTCCCTATATTTTCTGCAG
	PdMODD2-RT	GATCAAGCAGTGGTGGAGAAG	CCTCCCTCTATTTTCTGCAG
	PdMODD3-RT	TTTGAGAGCCAGCCACTT	CCATTCTCAGTGACTGTAAG
	actin	CATCCAGGCTGTCCTTTCCC	AACGAAGGATGGCGTGTGG
pBI121-PdMODD-GFP 融合表达载体构建 construction of pBI121- PdMODD-GFP fusion expression vector	PdMODD1-GFP	CTCTAGACTGGTACCCA TGGGAGAAGCAAACG	ACTAGTCAGTCGACCCGGTTC AAAGGACGGACAAG
	PdMODD2-GFP	CTCTAGACTGGTACCCATG GGAGAAACAAACG	ACTAGTCAGTCGACCCAAC ACAAAGGACGGACCAG
	PdMODD3-GFP	CTCTAGACTGGTACCCA TGGCACAAACAGAG	ACTAGTCAGTCGACCCGGA AAAAAGGGAGAGGGGTG

基因 gene	ORF 长度/bp ORF length	氨基酸数目 number of amino acid	分子质量/ ku relative molecular weight	理论等电点 pI	亚细胞定位 subcellular localization
PdMODD1	1 065	354	38.83	7.66	细胞核nucleus
PdMODD2	1 065	354	38.71	8.20	细胞核nucleus
PdMODD3	1 074	357	39.56	6.05	细胞核nucleus

Cloning and expression characteristics of PdMODD genes in Populus × euramericana

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 8

References 30

Related Articles 6

Recommended Articles

Metrics

Comments

Author

Reader

Reviewer

[1]	JIA Zhanhui, JIA Xiaodong, XU Mengyang, MO Zhenghai, ZHAI Min, XUAN Jiping, ZHANG Jiyu, WANG Gang, WANG Tao, GUO Zhongren. Cloning and expression analyses of the key enzyme gene of procyanidins biosynthesis in pecan (Carya illinoinensis) [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2022, 46(5): 49-57.
[2]	ZHU Youpeng, LIU Hongli, HAN Changzhi. Prediction of secretory protein in walnut bacterial black spot pathogen [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2019, 43(03): 17-22.
[3]	YANG Jie, SUN Lu, WANG Siyao, LI Ying, ZHAI Rui, LIN Xiangyu, ZHAN Yaguang, YIN Jing. Tissue specificity and hormone induced expression of three cytochrome P450 genes from Betula platyphylla Suk [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2018, 42(06): 27-34.
[4]	LONG Xiangyu, HE Bin, CAO Bing, LIANG Qifu, FANG Yongjun, YANG Jianghua, TANG Chaorong. Molecular cloning and expression analysis of HbF2KP from Hevea brasiliensis [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2014, 38(04): 23-28.
[5]	ZHANG Hongjuan, TAN Biyue, CAO Fuliang. Cloning and characterization of the geranylgeranyl pyrophosphate synthase gene from Ginkgo biloba Linn [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2013, 37(04): 8-12.
[6]	WU Xiaoyu,HU Shanglian, CAO Ying,LU Xueqin, REN Peng, ZHOU Meijuan, LI Xiaorui. Cloning of CCoAOMT gene in Neosinocalamus affinis and its bioinformatics analysis [J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2012, 36(03): 17-22.