
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.
Cloning and expression pattern analysis of NAC genes in Salix
【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.
[1] |
|
[2] |
|
[3] |
|
[4] |
|
[5] |
|
[6] |
|
[7] |
|
[8] |
|
[9] |
|
[10] |
|
[11] |
|
[12] |
|
[13] |
|
[14] |
|
[15] |
|
[16] |
|
[17] |
|
[18] |
|
[19] |
|
[20] |
|
[21] |
|
[22] |
|
[23] |
|
[24] |
|
[25] |
|
[26] |
|
[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.
|
[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.
|
[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.
|
[30] |
|
[31] |
|
[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.
|
[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.
|
[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.
|
[35] |
|
/
〈 |
|
〉 |