睡莲NAC转录因子的鉴定及其表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-0642

摘要/Abstract

摘要：

目的从睡莲基因组中鉴定NAC成员，并对其进化历程以及在不同组织和ABA胁迫下的表达模式进行分析，为睡莲NAC家族的分子进化提供理论基础，同时为睡莲NAC成员调控花器官形态建成和响应逆境胁迫提供候选基因。方法利用全基因组方法分析睡莲NAC家族成员的保守基序、基因结构、系统进化、染色体定位、基因复制事件、启动子顺式调控元件，并使用转录组和RT-qPCR对睡莲NAC家族成员在不同组织和ABA胁迫下的表达模式进行分析。结果从睡莲基因组中筛选出64个NAC成员，可分为15个亚家族。NcNAC成员不均匀地分布在睡莲14条染色体上，大部分成员定位于1、2、9和11号染色体上。睡莲NAC家族的片段复制事件与睡莲NAC家族的扩增紧密联系。NcNAC11、NcNAC43、NcNAC36和NcNAC52在花器官间的表达量较高，推断这些基因可能参与睡莲花器官的形态建成。NcNAC成员启动子区含有大量与胁迫响应相关的顺式调控元件，其中启动子区含有ABA响应元件最多的NcNAC07、NcNAC08、NcNAC34和NcNAC43能够响应ABA胁迫，表明NcNAC07、NcNAC08、NcNAC34和NcNAC43可能与睡莲的抗逆胁迫有关。结论在睡莲中有64个NcNAC成员，可以将其分为15个亚家族，NcNAC11、NcNAC43、NcNAC36和NcNAC52在花器官中的表达量较高，NcNAC07、NcNAC08、NcNAC34和NcNAC43能够响应ABA胁迫。

关键词: 睡莲, NAC家族, 转录因子, 分子进化, 表达分析

Abstract:

Objective This study aims to identify the NAC members in waterlily (Nymphaea colorata), and to investigate their evolutionary history and expressions under different tissues and ABA treatment. The current study not only could provide a key theoretical basis on the molecular evolution of the waterlily NAC family, but also could obtain some important candidate genes about regulating floral organ morphogenesis and stress response in waterlily. Method The whole-genome methods were applied to analyze the conserved motifs, gene structures, phylogenetic evolutions, chromosomal locations, gene duplication events, and cis-regulatory elements in the promoter regions of the waterlily NAC family. The expression levels of the waterlily NAC members under different tissues and ABA treatment were performed through the transcriptome and RT-qPCR. Result In this study, 64 NAC members screened from waterlily were divided into 15 subfamilies. The NcNAC members were unevenly distributed on the waterlily's 14 chromosomes, with most members located on chromosomes 1, 2, 9, and 11. Segmental duplication events were prevalent in the expansion of the NcNAC members. Meanwhile, NcNAC11, NcNAC43, NcNAC36 and NcNAC52 were highly expressed in floral organs. Thus, NcNAC11, NcNAC43, NcNAC36 and NcNAC52 may regulate the flower growth and development in waterlily. In addition, numerous stress-responsive regulatory elements were found in the promoter regions of the NcNAC members. NcNAC07, NcNAC08, NcNAC34, and NcNAC43 had the most ABA responsive elements in their promoter regions, and these NcNAC members responded to ABA stress, indicating that NcNAC07, NcNAC08, NcNAC34, and NcNAC43 might be related to stress resistance. Conclusion There are 64 NcNAC members in waterlily, and these NcNACs could be classified into 15 subfamilies. NcNAC11, NcNAC43, NcNAC36 and NcNAC52 have the highest expression in the floral-related tissues, and NcNAC07, NcNAC08, NcNAC34, and NcNAC43 may respond to ABA stress.

Key words: Nymphaea colorata, NAC family, transcription factor, molecular evolution, expression analysis

钱政毅, 吴绍芳, 曹舒怡, 宋雅欣, 潘鑫峰, 李兆伟, 范凯. 睡莲NAC转录因子的鉴定及其表达分析[J]. 生物技术通报, 2025, 41(2): 234-247.

QIAN Zheng-yi, WU Shao-fang, CAO Shu-yi, SONG Ya-xin, PAN Xin-feng, LI Zhao-wei, FAN Kai. Identification of the NAC Transcription Factors in Nymphaea colorata and Their Expression Analysis[J]. Biotechnology Bulletin, 2025, 41(2): 234-247.

图/表 10

表1 本试验使用的RT-qPCR引物序列

Table 1 Primer sequences of RT-qPCR in this study

基因名称Gene name	正向引物Forward primer （5′-3′）	反向引物Reverse primer （5′-3′）
NcUbi	CCTGTATTGCCTGATG	GACGAGATGGTGGAGT
NcNAC11	CGGTGTTGACAAGACGGAGA	CGTCCTCTTCGATGGTAGGC
NcNAC07	CAGGAGAGGGAAGCACAAGG	GTTGCCGAAGACAAAGCCAG
NcNAC08	GCTTCAGGTTCCATCCGACA	TTGGCACTGAAATTGGCTGC
NcNAC33	GGATGAGTACCGTCTCGCAG	CGTCCAAGTCGTCCAGTGAA
NcNAC34	TCCGGTTACTGGAAAGCCAC	ACTCGTGCATGATCCAGTCC
NcNAC36	CCTACAGCTGCACCCTTCAA	GGTGATCTATCAGTGCCGGG
NcNAC43	GGTGACAGCAGGAACGAAGA	CGCTGGTGATGATGCTTTCG
NcNAC52	ACTCTACCTGCATCGGTCCT	GGGATGAATGTGAAGGGGCA
NcNAC56	ATTTGCCTGGTGATGGGGAG	ACAACTTGGTTGCACCTGGA

图1 拟南芥和睡莲NAC家族的系统发育分析

Fig. 1 Phylogenetic analysis of the NAC family in A. thaliana and N. colorata

图2 睡莲中NAC家族进化分析（A）及其保守基序（B）与基因结构（C）的分布

Fig. 2 Phylogenetic analysis (A) and distribution of conserved motif (B) and gene structure (C) in the NAC family of N. colorata

图3 睡莲NAC成员的染色体分布

Fig. 3 Chromosomal distribution of NAC members in N. colorata

图4 睡莲NAC复制基因的共线性分析和Ks分布A：NcNAC复制基因的共线性分析；B：NcNAC复制基因的亚家族分布；C：NcNAC复制基因的Ks分布，箭头所指为Ks峰值

Fig. 4 Syntenic analysis and Ks distribution of the duplicated NAC members in N. colorataA: Syntenic analysis of the duplicated NcNACs. B: Subfamily distribution of the duplicated NcNACs. C: Ks distribution of the duplicated NcNACs, and the arrow points to peak value of Ks

图5 NcNAC成员启动子区域的胁迫响应调控元件分析

Fig. 5 Stress-responsive regulatory elements in the promoter regions of NcNAC members

图6 利用转录组数据分析不同组织中NcNAC成员的表达模式

Fig. 6 Expression profiles of NcNAC members in different tissues through the transcriptome data

图7 利用RT-qPCR方法分析不同组织中NcNAC成员的表达模式不同字母表示不同处理间在0.05水平差异显著。下同

Fig. 7 Expression profiles of NcNAC members in different tissues by RT-qPCR methodDifferent letters indicate the significant differences among different treatments at the 0.05 level. The same below

图8 睡莲NAC成员网络关系图

Fig. 8 Relationship network of the NcNAC members in N. colorata

图9 睡莲NcNAC成员在ABA胁迫下的表达情况

Fig. 9 Expressions of the NcNAC members in N. colorata under ABA stress

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