Cloning and expression pattern analysis of NAC genes in Salix

TIAN Xueyao, ZHOU Jie, WANG Baosong, HE Kaiyue, HE Xudong

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2020, Vol. 44 ›› Issue (1) : 119-124.

PDF(2718 KB)
PDF(2718 KB)
JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2020, Vol. 44 ›› Issue (1) : 119-124. DOI: 10.3969/j.issn.1000-2006.201905031

Cloning and expression pattern analysis of NAC genes in Salix

Author information +
History +

Abstract

【Objective】 NAC transcription factors play an important role in regulating plant growth, development and response to stress tolerance. Studying the sequence, structure, evolutionary relationship and expression pattern of NAC transcription factor gene family related with stress tolerance in willow will have important theoretical significance for understanding the molecular mechanism underlying stress tolerance, which in turn will provide a theoretical basis for molecular assisted breeding in future.【Method】 Two members of the NAC gene family were cloned based on the RNA-seq data of the Salix × jiangsuensis ‘J2345’ variety. The gene structure, protein properties and gene phylogeny were analyzed by using bioinformatics. The tissue specific expression in leaf and root, as well as the expression patterns under different types of stress was detected by quantitative real time PCR. 【Result】 The two NAC transcription factors cloned from the leaves of Salix × jiangsuensis ‘J2345’ variety were named SlNAC1 and SlNAC2. Bioinformatics analysis results showed that the sequence lengths of SlNAC1 and SlNAC2 were 1 126 bp and 1 139 bp, encoding proteins with 343 and 291 amino acid residues, respectively. The molecular weight of the proteins expressed from the two SlNAC1 and SlNAC2 genes were 40.1 ku and 42.3 ku, respectively, and both were stable, hydrophilic proteins. The SlNAC1 gene was located in nucleus while the SlNAC2 gene was located in chloroplast. Sequence alignment showed that both the genes contained typical NAM domains as well as A, B, C, D and E sub-domains, which shared the LPPG, YPNG and DEE conservative motifs and NAC suppression domains. Phylogenetic analysis demonstrated that SlNAC1 and SlNAC2 shared the highest homology with the genes from Manihot esculenta and Solanum melongena, respectively. RT-PCR results showed that both SlNAC1 and SlNAC2 were expressed in leaves and not in roots. The results of qRT-PCR showed that SlNAC1 was significantly upregulated after 24 hours of abscisic acid (ABA) and gibberellins (GA) treatments. SlNAC2 was significantly upregulated after exposure to polyethylene glycol (PEG), ABA and GA stresses. 【Conclusion】 The transcription factor SlNAC1 can be induced by induced by ABA and GA, and could stably express under abiotic stress. SlNAC2 can be induced by PEG, ABA and GA rather than ethrel (ETH), at a significantly higher expression level than SlNAC1. Therefore, we speculated that SlNAC1 and SlNAC2 were involved in GA and ABA signal transduction, but not in th e ETH signaling pathway.

Key words

Salix / NAC gene family / gene cloning / gene expression

Cite this article

Download Citations
TIAN Xueyao , ZHOU Jie , WANG Baosong , et al . Cloning and expression pattern analysis of NAC genes in Salix[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2020, 44(1): 119-124 https://doi.org/10.3969/j.issn.1000-2006.201905031

References

[1]
SINGH K, FOLEY R C, ONATE-SANCHEZ L. Transcription factors in plant defense and stress responses[J]. Current Opinion in Plant Biology, 2002, 5(5):430-436. DOI: 10.1016/s1369-5266(02)00289-3.
[2]
ERNST H A, OLSEN N A, 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. DOI: 10.1038/sj.embor.7400093.
[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. DOI: 10.1023/a:1016028530943.
[4]
OOKA H, SATOH K, DOI K, et al. Comprehensive analysis of NAC family genes in Oryza sativa and Arabidopsis thaliana[J]. DNA Research, 2003, 10(6):239-247. DOI: 10.1093/dnares/10.6.239.
[5]
SHEN H, YIN Y B, CHEN F, et al. A bioinformatic analysis of NAC genes for plant cell wall development in relation to lignocellulosic bioenergy production[J]. BioEnergy Research, 2009, 2(4):217-232. DOI: 10.1007/s 12155-009-9047-9.
[6]
PURANIK S, SAHU P P, SRIVASTAVA P S, et al. NAC proteins: regulation and role in stress tolerance[J]. Trends in Plant Science, 2012, 17(6):369-381. DOI: 10.1016/j.tplants.2012.02.004.
[7]
JENSEN M K, SKRIVER K. NAC transcription factor gene regulatory and protein-protein interaction networks in plant stress responses and senescence[J]. IUBMB Life, 2014, 66(3):156-166. DOI: 10.1002/iub.1256.
[8]
XUE G P, WAY H M, RICHARDSON T, et al. Overexpression of TaNAC69 leads to enhanced transcript levels of stress up-regulated genes and dehydration tolerance in bread wheat[J]. Molecular Plant, 2011, 4(4):697-712. DOI: 10.1093/mp/ssr013.
[9]
YANG R C, DENG C T, OUYANG B, et al. Molecular analysis of two salt-responsive NAC-family genes and their expression analysis in tomato[J]. Molecular Biology Reports, 2011, 38(2):857-863. DOI: 10.1007/ s11033-010-0177-0.
[10]
MAO X G, ZHANG H Y, QIAN X Y, et al. TaNAC2, a NAC-type wheat transcription factor conferring enhanced multiple abiotic stress tolerances in Arabidopsis[J]. Journal of Experimental Botany, 2012, 63(8):2933-2946. DOI: 10.1093/jxb/err462.
[11]
GAO F, XIONG A S, PENG R H, et al. OsNAC52, a rice NAC transcription factor, potentially responds to ABA and confers drought tolerance in transgenic plants[J]. Plant Cell, Tissue and Organ Culture (PCTOC), 2010, 100(3):255-262. DOI: 10.1007/s11240-009-9640-9.
[12]
KIM S Y, KIM S G, KIM Y S, et al. Exploring membrane-associated NAC transcription factors in Arabidopsis: implications for membrane biology in genome regulation[J]. Nucleic Acids Research, 2007, 35(1):203-213. DOI: 10.1093/nar/gkl1068.
[13]
SHAN W, KUANG J F, CHEN L, et al. Molecular characterization of banana NAC transcription factors and their interactions with ethylene signalling component EIL during fruit ripening[J]. Journal of Experimental Botany, 2012, 63(14):5171-5187. DOI: 10.1093/jxb/ers178.
[14]
DELESSERT C, KAZAN K, WILSON I W, et al. The transcription factor ATAF2 represses the expression of pathogenesis-related genes in Arabidopsis[J]. The Plant Journal, 2005, 43(5):745-757. DOI: 10.1111/j.1365-313x.2005.02488.x.
[15]
YOSHII M, YAMAZAKI M, RAKWAL R, et al. The NAC transcription factor RIM1 of rice is a new regulator of jasmonate signaling[J]. The Plant Journal, 2010, 61(5):804-815. DOI: 10.1111/j.1365-313x.2009.04107.x.
[16]
KIM S G, LEE A K, YOON H K, et al. A membrane-bound NAC transcription factor NTL8 regulates gibberellic acid-mediated salt signaling in Arabidopsis seed germination[J]. The Plant Journal, 2008, 55(1):77-88. DOI: 10.1111/j.1365-313x.2008.03493.x.
[17]
RIECHMANN J L, HEARD J, MARTIN G, et al. Arabidopsis transcription factors: genome-wide comparative analysis among eukaryotes[J]. Science, 2000, 290(5499):2105-2110. DOI: 10.1126/science.290.5499.2105.
[18]
HAN X J, HE G, ZHAO S T, et al. Expression analysis of two NAC transcription factors PtNAC068 and PtNAC154 from Poplar[J]. Plant Molecular Biology Reporter, 2012, 30(2):370-378. DOI: 10.1007/s11105-011-0350-1.
[19]
OLSEN A N, ERNST H A, LEGGIO L L, et al. NAC transcription factors: structurally distinct, functionally diverse[J]. Trends in Plant Science, 2005, 10(2):79-87. DOI: 10.1016/j.tplants.2004.12.010.
[20]
RUSHTON P J, BOKOWIEC M T, HAN S C, et al. Tobacco transcription factors: novel insights into transcriptional regulation in the Solanaceae[J]. Plant Physiology, 2008, 147(1):280-295. DOI: 10.1104/pp.107.114041.
[21]
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. DOI: 10.1016/j.gene.2010.06.008.
[22]
SHAH S T, PANG C Y, HUSSAIN A, et al. Molecular cloning and functional analysis of NAC family genes associated with leaf senescence and stresses inGossypium hirsutum L.[J]. Plant Cell, Tissue and Organ Culture (PCTOC), 2014, 117(2):167-186. DOI: 10.1007/s11240-014-0430-7.
[23]
ZHANG C, ZHANG L, ZHANG S, et al. Global analysis of gene expression profiles in physic nut (Jatropha curcas L.) seedlings exposed to drought stress[J]. BMC Plant Biology, 2015, 15(1):17. DOI: 10.1186/s12870-014-0397-x.
[24]
HU R B, QI G, 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. DOI: 10.1186/1471-2229-10-145.
[25]
XU Z Y, GONGBUZHA X I, WANG C Y, et al. Wheat NAC transcription factorTaNAC29 is involved in response to salt stress[J]. Plant Physiology and Biochemistry, 2015, 96:356-363. DOI: 10.1016/j.plaphy.2015.08.013.
[26]
JEONG J S, KIM Y S, REDILLAS M C F R, et al. OsNAC5 overexpression enlarges root diameter in rice plants leading to enhanced drought tolerance and increased grain yield in the field[J]. Plant Biotechnology Journal, 2013, 11(1):101-114. DOI: 10.1111/pbi.12011.
[27]
刘旭, 李玲. 花生NAC转录因子AhNAC2和AhNAC3的克隆及转录特征[J]. 作物学报, 2009, 35(3):541-545.
LIU X, LI L. Cloning and characterization of the NAC-like geneAhNAC2 and AhNAC3 in Peanut [J]. Acta Agronomica Sinica, 2009, 35(3):541-545. DOI: 10.3724/SP.J.1006.2009.00541.
[28]
唐宽刚, 任美艳, 张文君, 等. 沙冬青AmNAC6基因的克隆与功能初步分析[J]. 植物科学学报, 2018, 36(5):705-712.
TANG K G, REN M Y, ZHANG W J, et al. Cloning and preliminary functional analysis of AmNAC6 from Ammopiptanthus mongolicus [J]. Plant Science Journal, 2018, 36(5):705-712. DOI: 10.11913/PSJ.2095-0837.2018.50705.
[29]
武肖琦, 冯涛, 刘艳军, 等. 桃PpNAC72的克隆及表达分析[J]. 生物技术通报, 2019, 35(6):1-8.
WU X Q, FENG T, LIU Y J, et al. Cloning and expression analysis of the PpNAC72 in peach [J]. Biotechnology Bulletin, 2019, 35(6):1-8. DOI: 10.13560/j.cnki.biotech.bu11.1985.2018-0891.
[30]
TIAN X Y, ZHENG J W, JIAO Z Y, et al. Transcriptome sequencing and EST-SSR marker development in Salix babylonica and S. suchowensis[J]. Tree Genetics & Genomes, 2019, 15:9. DOI: 10.1007/s11295-018-1315-4.
[31]
ZHENG X N, CHEN B, LU G J, et al. Overexpression of a NAC transcription factor enhances rice drought and salt tolerance[J]. Biochemical and Biophysical Research Communications, 2009, 379(4):985-989. DOI: 10.1016/j.bbrc.2008.12.163.
[32]
邵帅, 徐岭贤, 王绍辉, 等. 茄子SmNAC1基因的克隆与表达分析[J]. 园艺学报, 2014, 41(5):975-984.
SHAO S, XU L X, WANG S H, et al. Cloning and expression analysis of SmNAC1 in Solanum melongena [J]. Acta Horticulturae Sinica, 2014, 41(5):975-984. DOI: 10.16420/j.issn.0513-353x.2014.05.020.
[33]
姜秀明, 牛义岭, 徐向阳. 番茄NAC基因家族的系统进化及表达分析[J]. 分子植物育种, 2016, 14(8):1948-1964.
JIANG X M, NIU Y L, XU X Y. Phylogenetic evolution and expression analysis of NAC gene family in tomato (Solanum lycopersicum) [J]. Molecular Plant Breeding, 2006, 14(8):1948-1964. DOI: 10.13271/j.mpb.014.001948.
[34]
丁泽红, 颜彦, 付莉莉, 等. 木薯NAC转录因子Rd26基因克隆及表达[J]. 南方农业学报, 2016, 47(11):1822-1826.
DING Z H, YAN Y, FU L L, et al. Clone and expression of NAC transcription factor Rd26 gene from Manihot esculenta Crantz [J]. Journal of Southern Agriculture, 2016, 47(11):1822-1826. DOI: 10.3969/jissn.2095-1191.2016.11.1822.
[35]
FUJITA M, FUJITA Y, MARUYAMA K, et al. A dehydration-induced NAC protein,RD26, is involved in a novel ABA-dependent stress-signaling pathway[J]. The Plant Journal, 2004, 39(6):863-876. DOI: 10.1111/ j.1365-313x.2004.02171.x.

RIGHTS & PERMISSIONS

Copyright reserved © 2020
PDF(2718 KB)

Accesses

Citation

Detail

Sections
Recommended
The full text is translated into English by AI, aiming to facilitate reading and comprehension. The core content is subject to the explanation in Chinese.

/