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Article

Identification and Expression Analysis of the Ethylene Response Factor Gene Family in Tea Plant (Camellia sinensis)

by
Tao Zhang
1,
Ju Cai
1,
Sisi Wang
1,
Litang Lv
2,
Dandan Yuan
1,
Xiaofang Zeng
1,
Jianrong Li
1 and
Yan Li
1,*
1
The Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), College of Life Sciences, Institute of Agro-Bioengineering, Guizhou University, Guiyang 550025, China
2
College of Tea Sciences, Guizhou University, Guiyang 550025, China
*
Author to whom correspondence should be addressed.
Agronomy 2023, 13(7), 1900; https://doi.org/10.3390/agronomy13071900
Submission received: 16 June 2023 / Revised: 13 July 2023 / Accepted: 17 July 2023 / Published: 19 July 2023
(This article belongs to the Special Issue Decrypting the Mechanism of Protein Transcription in Plant Stress Resistance)
Browse Figures
Review Reports Versions Notes

Abstract

:
The ERF gene family is widely present in plants and has crucial regulatory importance in plant seed development, organ morphogenesis, the synthesis of secondary metabolites, and coping with abiotic stresses such as cold and drought. In this study, 90 members of CsERF were screened by bioinformatics tools analysis and named CsERF1–CsERF90. Their molecular characteristics and systematic evolution were studied, and the tissue expression characteristics of CSERF genes and the composition of promoter cis-acting elements were predicted. The results showed that 81 proteins encoded by CsERF genes had conserved motifs 1, 2, and 3, while 64 members possessed other motifs. The theoretical isoelectric point was between 4.49 and 10.24, and 85 members constituted unstable proteins, while the rest were stable proteins. Subcellular localization predicted that 77 members were in the nucleus, 8 were in the chloroplasts, and 5 were in the mitochondria. The promoter sequence of CsERFs was found to include not only cis-acting elements related to hormone regulation, such as gibberellin (41), methyl jasmonate (110), and abscisic acid (185), but also cis-acting elements involved in low-temperature response (56) and light response (22), indicating that CsERFs have a key role in plant growth and abiotic stress. Phylogenetic analysis of tea plant and Arabidopsis thaliana ERF gene families showed that the tea plant ERF gene families could be divided into six groups, with B3 having 29 members at most and B1 having only 3 members at least. The phylogenetic tree constructed using only the CsERF genes is also divided into six groups, with slightly different but minimal differences in members. Of the 90 tea plant ERF members, 85 were located on 15 chromosomes, whereas 5 were not located on chromosomes. The collinearity analysis showed that there were 41 homologous gene pairs among the CsERFs, and these homologous gene pairs may have the same function. According to the expression of CsERFs in cold-stressed tea plant and in different tissues, 90 CsERF genes played their respective roles in different tissues and stages to regulate plant growth, and some of them participated in the process of cold stress tolerance. This study provides a theoretical foundation for the study of tea plant growth and development and low-temperature resistance.

1. Introduction

Tea plant (Camellia sinensis (L.) O. Ktze) is a very important cash crop [1]. It is the oldest and most popular nonalcoholic beverage in the world, and was first discovered and utilized in China [2]. Tea plant contains many beneficial ingredients, such as tea polyphenols, alkaloids, amino acids, and vitamins, which can effectively reduce cholesterol and hypertension levels and reduce the risk of stroke [3]. Chill is a vital environmental element that affects the growth, yield, and regional distribution of tea plant. Chill freezing injury often occurs during the process of tea planting in China, especially in late spring, and is extremely harmful to early-growing tea plant varieties [4]. Many studies have suggested that when plants are subjected to cold stress, they will improve their tolerance by regulating the expression of a series of genes. Many transcription factors (TFs), including ERF, WRKY, and MYB, are important regulatory factors related to cold stress [5,6].
The ETHYLENE RESPONSE FACTOR (ERF) gene family is one of the subfamilies of the AP2/ERF (APETALA2/ETHYLENE RESPONSE FACTOR) gene family, which is one of the biggest transcription factor families in plants. It contains an AP2 conserved domain composed of 60–70 amino acids [7]. In addition to the ERF gene family, the AP2/ERF gene family contains the AP2, DREB, RAV, and Soloist subfamilies [8]. Since the first AP2/ERF gene family member was found in A. thaliana in 1994 [9], the ERF gene family has been identified in many plants; for example, in Cucumis sativus [10], Glycine max [11], Ipomoea batatas [12], Panax ginseng [13], Fragaria vesca [14], and Gynostemma pentaphyllum [15]. In 2006, Shigyo et al. found homologous proteins of the AP2/ERF family in green algae, mosses, and gymnosperms [16], and Magnani and Balaji et al. found similar sequences in bacteria and viruses in 2004 and 2005, respectively [17,18]. It has been known for a long time that the ERF gene family is involved in many processes, such as plant growth and development [10,19] and abiotic or biotic stress [20,21]. Samriti et al. found that some of the Pisum sativum ERF gene family could resist cold stress [22]. Klay et al. also found a similar function in Solanum lycopersicum [23]. A motif is a characteristic short sequence, which is considered as an insular sequence that has biological functions. It may include specific binding sites and a mutual sequence segment involving a particular biological process. In addition to their own conserved domain, ERF genes have conserved motifs that play a key role in the transcriptional regulation of ERF [24]. For example, the ERA (ERF-associated amphiphilic repression) motifs, LxLxLx and DLNxxP, can directly affect the function of ERF transcriptional regulation [25]. By extracting the motif sequence, we can predict potential binding sites, which will help in the study of the function of different genes.
The phylogenetic relationships between different genes can be observed using a phylogenetic tree. In 2006, Nakano et al. [7] studied the phylogenetic relationships between A. thaliana and Oryza sativa ERF gene families, and they divided the ERF gene families of the two species into 6 groups and 10 groups, respectively. In 2008, Zhang et al. [11] divided the Glycine max ERF gene family into six groups based on phylogenetic analysis. Xing et al. [26] divided the ERF gene family in Zingiber officinale into six groups in 2021. At present, most studies have divided the ERF gene family into six groups in phylogenetic analyses, such as in Triticum durum [27] and Solanum melongena [28]. Their classification elements are to refer to the classification standard of A. thaliana [7].
cis-acting elements are sequences that exist in the flanking sequences of genes and can influence gene expression. cis-acting elements, such as promoters, enhancers, regulatory sequences, and derivable elements, are involved in gene expression regulation. They themselves do not encode any proteins, only providing a functional site that requires interaction with transacting factors [29]. The ERF gene family has many cis-acting elements that can participate in various growth and development processes and abiotic stresses in plants, such as ABRE, LTR, W box, and DRE, and also participate in the expression of various genes in plants [30].
Collinearity refers to the gene linkage in genetics. Through the analysis of gene collinearity, we can determine the genes in a species that have obviously expanded or contracted, and these changes may be related to some enhanced or weakened biological molecular functions of the species [31]. The nonsynonymous substitution rate (Ka) to synonymous substitution rate (Ks) is diffusely used as a pointer of different gene selection pressures. If Ka/KS is greater than 1, it is believed that there is a positive selection effect, while if Ka/Ks = 1, it is believed that there is neutral selection, and if Ka/KS is less than 1, it is believed that there is a decontamination chose effect [32]. Whole-genome duplication (WGD) and polyploidy are considered as the sources of plant evolution [33]. The ERF gene family was amplified in a WGD event, and studies have found that segmental duplication also had an influence on the amplification of the Gossypium hirsutum ERF gene family [34], which is consistent with that discovered in Rosa chinensis [35].
Many studies have shown that the change in ERF gene expression can change the tolerance of plants to cold. For example, Sun et al. found that the ERF092 gene in Amur gape can enhance the expression of the WRKY33 gene under cold stress, and indirectly enhance its cold resistance [36]. Zhang et al. found that the ERF39 gene in Eriobotrya japonica loquat can enhance the expression of the lignin gene under cold stress to enhance cold resistance [37]. Zhang et al. found that the ERF2 gene in S. lycopersicum can enhance the cold tolerance of plants by increasing the expression of genes containing GCC box and DRE/CRT [38]. In this study, bioinformatics analysis was used to identify ERF gene family members in tea plant and analyze their molecular characteristics, systematic evolution, and resistance to low-temperature stress, in order to provide a theoretical foundation for the study of tea plant growth, development, and low-temperature resistance.

2. Materials and Methods

2.1. Identification of ERF Gene Family Members in Tea Plant

First, the protein sequences of Arabidopsis AP2/ERF gene family members were downloaded from the Arabidopsis database (https://www.arabidopsis.org/ accessed on 4 September 2022) [39], and all the protein sequences of tea plant were downloaded from the tea plant database (http://tpdb.shengxin.ren/ accessed on 4 September 2022) [40], and the target sequences were preliminarily screened out by TBtools software version 0.665 (E-value: 1 × 10−5) [41]. Then, the HMM of the AP2 domain (PF00847) in the Pfam database (https://www.ebi.ac.uk/interpro/entry/pfam/#table accessed on 5 September 2022) [42] was downloaded and used for a search in hmmer (3.0) to obtain the target sequence [43]. The target sequences obtained by the above two methods were summarized into tea plant AP2/ERF gene family members, and then the above sequences were BLASTed in NCBI (https://www.ncbi.nlm.nih.gov/ accessed on 7 September 2022).
The conservative domain of the sequence screened after BLAST was found in the CDD database (https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi accessed on 8 September 2022) (default parameters), and the sequence containing only one AP2 domain was reserved [44]. Then, it was combined with 65 AtERFs [45] to construct a phylogenetic tree in MEGA (default parameters), and then finally, the CsERF members were screened.

2.2. Physicochemical Properties of ERF Gene Family Players in Tea Plant

The online tool ProtParam (https://web.expaty.org/protparam/ accessed on 25 October 2022) was used to predict the length, molecular weight, theoretical isoelectric point (pI), instability coefficient, and average coefficient of hydrophilicity of the protein of the ERF gene family in tea plant [46]. Online tools such as TMHMM [47] (https://dtu.biolib.com/DeepTMHMM accessed on 27 October 2022), WoLF PSORT [48] (https://wolfpsort.hgc.jp/ accessed on 28 October 2022), Cell-Ploc [49] (http://www.csbio.sjtu.edu.cn/bioinf/Cell-PLoc-2/ accessed on 10 July 2023), and SOPMA [50] (https://npsa-prabi.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_sopma.html accessed on 29 October 2022) were used to predict the transmembrane characteristics, subcellular localization, and secondary structure of the CsERFs. The CDD database (https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi accessed on 3 November 2022) in NCBI was used to predict the position of its conserved domain [44].

2.3. Visualization of Gene Structure, Domain, and Conserved Motif of ERF Gene Family Members in Tea Plant

The information relating to the introns and exons of the CsERF database (http://tpdb.shengxin.ren/ accessed on 4 September 2022) was obtained, and the conservative domain information of the CsERFs was obtained from the CDD database (https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi accessed on 3 November 2022) in NCBI [44]. The MEME (https://meme-suite.org/meme/tools/meme accessed on 10 November 2022) tool was used to predict its conservative motif online [51], and it was visualized with TBtools software version 0.665 [41].

2.4. Phylogenetic Analysis and Chromosome Mapping of ERF Gene Family Members in Tea Plant

The members of CsERFs and AtERFs were compared by ClustalX [52] software version 2.1, and used the neighbor-joining method of MEGA X software version 1.0, with Bootstrap of 1000 to constructe the phylogenetic tree [53]. Then only CsERFs genes were used to construct phylogenetic trees by neighbor-joining method.
The position information of CsERFs was obtained from the tea plant database (http://tpdb.shengxin.ren/ accessed on 4 September 2022), and visualized by using TBtools software version 0.665 [41].

2.5. cis-Acting Elements of ERF Gene Family Members in Tea Plant

The promoter of the start codon 2000 bp upstream was separated from the genome file of the tea plant using TBtools software version 0.665, and then the cis-acting elements of the CsERFs were found using the online tool PlantCare (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/ accessed on 20 September 2022) [54] and then visualized using TBtools software version 0.665 [41].

2.6. Collinearity of ERF Gene Family Members in Tea Plant

The genome and annotation files were downloaded from the tea plant database [40] (http://tpdb.shengxin.ren/ accessed on 4 September 2022), and the collinearity among members was calculated using the MCScanX method. Mapping was carried out using TBtools software version 0.665 [41].

2.7. Differences in the Expression of CsERF Gene Family Members

The expression levels (FPKM) of eight different tissues of tea plant (roots, mature leaves, stems, flowers, young leaves, old leaves, terminal buds, and fruits) and the expression levels of tea plant under cold stress were downloaded from the tea plant database (http://tpdb.shengxin.ren/ accessed on 4 September 2022) [40]. The expression levels (FPKM) were calculated with log2 (FPKM + 1) [55] and visualized with TBtools software version 0.665 [41].

3. Results

3.1. Identification and Physicochemical Property Analysis of CsERFs

Through identification and screening, 90 CsERFs were ultimately screened. The identified members were named CsERF1–CsERF90 (Table A1 in Appendix A). The quantity of AA encoded by the members of CsERF was 137–430, the protein molecular weights ranged from 15,234.30 to 47,271.43 D, and the theoretical pI ranged from 4.49 to 10.24. Protein hydrophilic and hydrophobic analysis showed that among the 90 CsERFs, the proteins with the weakest hydrophilicity obtained grand average of hydropathy (GRAVY) values of −0.200, while those with the strongest hydrophilicity obtained GRAVY scores of −1.186, all of which were hydrophilic proteins. The instability coefficient showed that most members of the CsERF gene family were variable proteins (instability index > 40), and only CsERF40, CsERF41, CsERF57, CsERF76, and CsERF77 were steady proteins (instability index < 40), among which CsERF57 obtained a value of only 27.54. The results of the transmembrane domain analysis revealed that 90 CsERF gene family members did not have a transmembrane domain. The predicted subcellular localization showed that 8 members, including CsERF28, CsERF50, CsERF60, CsERF67, CsERF73, CsERF75, CsERF80, and CsERF84, were located in the chloroplasts; CsERF42, CsERF46, CsERF57, CsERF79, and CsERF89 were located in the mitochondria; and the remaining 77 members were located in the nucleus. According to the secondary structure prediction, all CsERF members possessed no β-bridge, except for CsERF46 and CsERF71, and the other members had more than 40% random coils. Only two members, CsERF46 and CsERF59, had more α-helixes than random coils (Table A2).

3.2. Structural Composition and Conservative Motif Analysis of ERF Family Proteins in Tea Plant

The ERF protein only has a complete AP2 domain, and the position of the AP2 domain differs because of the diverse amino acid quantities of CsERF proteins (Table A1). Using TBtools software version 0.665, the evolutionary relationships of CsERF members, the location of the AP2 domain, and the locations of conservative motifs and protein-coding regions (CDSs) and noncoding regions (UTRs) were mapped (Figure 1). It was found that the tea plant ERF gene family possessed 2–7 motifs, and most of them (81) possessed motifs 1, 2, and 3, which indicates that the CsERFs have certain conservative motifs (Figure 2). In addition, 66 members also possessed other motifs, and members with similar motifs clustered together. These players with identical motifs may have identical capabilities. It was also found that the CsERFs had few noncoding regions, ranging from 0 to 3.

3.3. Phylogenetic Tree and Chromosome Mapping of CsERFs

In this research, 90 CsERFs and 65 AtERFs were compared using ClustalX software version 2.1, and then MEGA X software version 1.0 was used to construct a phylogenetic tree with only CsERF genes and both present (Figure 3 and Figure 4). Based on the classification of Sakuma et al., the ERF gene families were divided into six groups, B1 to B6, which were marked using different colors [8]. Among them, B1 possessed only 3 members of the CsERF gene family, while B3 possessed 29 members of the CsERF gene family. The phylogenetic tree constructed using only CsERFs is slightly different from it. In the CsERFs’ phylogenetic tree, CsERF11 was divided from B2 to B3, while CsERF7 and CsERF66 were divided from B6 to B5. Using TBtools software version 0.665, the CsERFs were mapped to tea plant chromosomes (Figure 5). Except for five members, namely CsERF5, CsERF19, CsERF29, CsERF40, and CsERF51, all the other 85 members were mapped to tea plant chromosomes, of which 15 members were located at chromosome 1 at most, and only 1 member was located at chromosome 3. This study found that there are 11 pairs of tandem repeat genes in CsERFs, which have been marked in blue in the figure (Figure 5).

3.4. cis-Acting Elements of ERF Gene Family Promoters in Tea Plant

By analyzing the cis-acting elements upwards of 2000 bp of the promoter of the CsERF gene family, it was found that almost all the CsERF gene family members possessed cis-acting elements regulated by hormones. Examples include abscisic acid response element (ABRE), salicylic acid response element (TCA-element), auxin response element (TGA-element), methyl jasmonate response element (TGACG-motif), and gibberellin response element (P-box). Some members also possessed light-responsive elements (3-AF1 binding site, AAAC-motif, ACE), low-temperature-responsive elements (LTR, DRE), and other stress-related elements (W box, MYB), while others possessed low-temperature-responsive functions, such as ABRE (Figure 6).

3.5. Collinearity Analysis of ERF Gene Family Members in Tea Plant

To explore the conservatism of CsERF gene family members during evolution, collinearity analysis of CsERF gene family players was carried out. It was found that 41 pairs of highlighted homologous genes existed among the CsERF gene family members (Figure 7) Furthermore, the values of Ka and Ks were calculated for these 41 pairs of highlighted homologous genes (Table 1). With the exception of CsERF13/CsERF33 and CsERF75/CsERF30, the value of Ka/Ks of the other 39 pairs of homologous genes was less than 1, which indicates that the 39 pairs of homologous genes evolved under great purifying selection or negative selection pressure [55]. The value of Ka/Ks of the other two pairs of highlighted genes was NaN, which may be due to synonymous mutations in most sites where synonymous mutations can occur.

3.6. Differences in Expression of ERF Gene Family Members in Tea Plant

To further explore the regulation of CsERFs on growth and low-temperature stress, the transcriptome data (FPKM) of eight tissues and cold-stressed plants obtained from the tea plant database were calculated and visualized (Figure 8 and Figure 9). It was found that the 90 ERF members could be divided into four categories: 22 members expressed only in individual tissues; 14 members had high expression in almost all tissues; 45 members had very low expression in almost all tissues; and the remaining 9 members had low expression quantities in the individual tissues.
However, the members of the old ERF gene family in tea plant could be separated into three categories under cold stress: 19 members had increased expression under cold stress; the expression quantity of 5 members decreased under cold stress; and the remaining 66 members did not respond to cold stress. Seventeen genes with upregulated expression under cold stress all possessed cis-acting elements (LTR, DRE, W box, MYB, and ABRE) related to cold stress, indicating that some CsERF members may participate in the process of resistance to cold stress.

4. Discussion

The AP2/ERF gene family is one of the maximal transcription factors in plants, and its members have multiple functions and exhibit a complex regulatory network. The ERF subfamily is its largest subfamily, and thus, it is difficult to select the ERF gene subfamily from the AP2/ERF gene family [56]. In A. thaliana and O. sativa, Nakano et al. found that they have 122 and 139 ERF family genes, respectively [7]. In Zea mays, Cheng et al. found that it has 105 ERF family genes [57]. In P. sativum, Jarambasa et al. found that it has 103 ERF family genes [58]. In F. vesca and Durio zibethinus, Su et al. and Khaksar et al. found that they have 57 and 63 ERF family genes, respectively [14,59]. In tea plant, Liu et al. identified 178 AP2/ERF family genes [60]. In this study, 90 ERF family genes were identified in tea plant. This shows that the number of ERF gene families varies greatly among different species.
Most studies have found that the majority of ERF gene family members are unstable and hydrophilic proteins. Chen et al. found that proteins encoded by members of the ERF gene family in Saccharum officinarum are hydrophilic proteins, most of which are unstable [61]. The 90 CsERFs identified in this research were all hydrophilic proteins, and most of them were unstable. In most studies, the theoretical pI values of ERF gene family members are very similar. Karanja et al. found that the theoretical pI values of the ERF gene family players of Raphanus sativus were between 4.46 and 10.1 [62]. Zhu et al. found that the theoretical pI values of the ERF gene family players of Pinus massoniana were between 4.63 and 11.61 [63], while Cao et al. found that the theoretical pI values of the ERF gene family members of Ammopiptanthus nanus were between 4.39 and 10.13 [64], which is very similar to the theoretical pI of the CsERFs obtained herein. Wu et al. discovered the AP2/ERF gene family in tea plant in 2015 [45] and also divided the CsERFs into six groups; however, only 45 ERF gene family members were identified. The present study identified more members, probably because of the different methods used. It was also found herein that the B3 class in the CsERFs had the largest number of members, which is consistent with the analysis of Wu et al. [45].
Studying the structural differences of gene family members is helpful for explaining the functional differentiation among multiple members of multiple species. It has been found that all members of the ERF gene family in G. hirsutum have motif 1 and motif 2, and some have motif 3 [34]. Motif 1 and motif 2 were also found in Glycine max [65]. Motif 1, motif 2, and motif 3 are relatively conserved motifs among the ERF members of tea plant, and are similar to—but not the same as—those of S. melongena [66]. This indicates that the ERF gene family is highly conserved, though it does possess some significant differences. At the same time, the present study also found that 66 CsERFs had other conserved motifs, which may be related to functional differentiation during evolution. Moreover, research has found that the ERF gene family in S. melongena [28] and Salvia miltiorrhiza [67] has few introns, generally 0–2, which is similar to the CsERFs found during this research.
Phylogenetic analysis can elucidate the evolutionary relationships among different species or different genes. Sakuma et al. [8] and Nakano et al. [7], respectively, separated the ERF gene family of A. thaliana into six groups in the phylogenetic tree, and Rao et al. [68] and Qian et al. [69] also separated the ERF gene family of Salix arbutifolia and Erianthus fulvus into six groups in the phylogenetic tree. In the present study, the phylogenetic tree indicated that the CsERF members and AtERF members also clustered into six categories. The self-expansion value in each group is relatively high, indicating that the genetic relationship of CsERF genes in each group is close. They may perform similar functions under certain conditions, among which B1, B2, and B3 include genes encoding proteins that specifically bind to GCC boxes, such as ERF-AT1G50640.1, ERF-AT3G16770.1, and ERF-AT3G23240.1 [8]. It is speculated that the products of these three categories may bind to GCC boxes or other related sequences.
Transcription factors can regulate gene expression by tying up cis-acting elements. A variety of cis-acting elements can induce or suppress the expression of genes in response to various biological or abiotic stress signals. Li et al. predicted the cis-acting elements of the S. melongena ERF gene family and found that its cis-acting elements were mainly light response elements, low-temperature induction elements, and phytohormone response elements [28]. Muhammad et al. found that the G. hirsutum ERF gene family also has these cis-acting elements [34]. In this study, various phytohormone response elements and lightresponse elements were found in the promoter region of ERF gene family in tea plant. It is noteworthy that this study also found that 73 members of ERF family genes in tea plant all contain cis-acting elements related to adversity stress such as LTR, DRE, and ABRE. It has been found that plants exhibit a trade-off between growth and defense (GDT) strategy; that is, when plants encounter biotic and abiotic stresses, they often sacrifice their growth to activate their defense systems. For example, treating plants with similar biotic stress substances that do not damage photosynthetic tissues can also delay their growth [70]. However, this study discovered many CsERFs with both resistance elements and growth hormone response elements. These genes may participate in the trade-off between defense and growth, and are important selection nodes in growth, development, and resistance.
Replication among members of a gene family promotes plant evolution, and collinearity can predict homologous gene sequences during evolution [71]. In R. chinensis, Li et al. found that the ERF gene family had 21 pairs of homologous genes, and its Ka/Ks value was less than 0.4, which confirmed that the R. chinensis ERF gene family has undergone purifying selection [35]. In the collinearity analysis of the CsERF gene family, 41 pairs of highlighted gene pairs were found, and the Ka/Ks count of 39 groups was ≤0.468, indicating that they had also gone through purifying selection during evolution [55], while the remaining two pairs of sites that might have synonymous mutations all mutated, signaling that the CsERF gene family was conserved during evolution. Studies have shown that tea plant have experienced a WGD [72], and WGD and replication play a significant role in ERF gene amplification in tea plant, as has also been found for Saccharum spontaneum, Brassica napus, and S. lycopersicum [30,73,74].
It has been found that the ERF gene family is constitutively expressed in many plants. In Z. officinale, Xing et al. found that some ERF family genes were expressed in different parts of plants [26]. In Cucumis melo, Ma et al. also found that some ERF genes were expressed in different parts of plants [75]. In tea plant, some ERF family genes discovered in this study are consistent with these species. In addition, a large number of studies show that many ERF family genes can be induced to expression. In Rhododendron simsii, Guo et al. found that 16 AP2/ERF genes were induced by cold [76]. In E. fulvus, Qian et al. found that 10 ERF genes were induced by cold [69]. In Z. mays, Cheng et al. found that one ERF gene was induced by salt, 2 ERF genes were induced by drought and 14 ERF genes were induced by cold [57]. In tea plant, Liu et al. found that 9 AP2/ERF genes were induced by light and 9 AP2/ERF genes were induced by cold. They mainly analyzed the AP2/ERF gene family in the spring bud break stage of tea plant and the expression pattern of tea plant in different periods of spring bud break. It provides candidate genes for molecular breeding. [60]. The present study focused on the identification of ERF genes related to low-temperature response in tea plant, and 19 ERF genes (CsERF1, CsERF12, CsERF24, CsERF34, and CsERF88, etc.) were identified as being induced by cold. This provides a basis for the follow-up, identifying resistance genes in tea plant. From this study, we can know that the ERF gene family of tea plant has a very wide range of biological functions. It can participate in plant growth and stress.

5. Conclusions

In this study, 90 members of the ERF gene family in tea plant were identified. Many CsERFs may participate in the growth and development of tea plant, and may also play an important role in cold resistance. The results of this study provide a theoretical basis for the functional study of ERF genes during tea plant growth and development and cold stress, and may also help in the screening of cold-resistant genes.

Author Contributions

Conceptualization, T.Z. and Y.L.; formal analysis, T.Z., J.C., S.W. and D.Y.; funding acquisition, Y.L.; investigation, J.C., S.W., D.Y. and Y.L.; methodology, T.Z. and Y.L.; supervision, L.L., X.Z., J.L. and Y.L.; validation, J.C., S.W. and D.Y.; visualization, T.Z.; writing—original draft, T.Z.; writing—review and editing, L.L., X.Z., J.L. and Y.L. All authors have read and agreed to the published version of the manuscript.

Funding

This is a key research project of Guizhou Provincial Science and Technology Projects (QKHJC-ZK [2023]ZD-006), the National Natural Science Foundation of China (32160722), the Open Project of Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Regions (Ministry of Education) (QJHKY-2022-367).

Data Availability Statement

The complete original data of this study can be obtained from the first author and the corresponding author.

Conflicts of Interest

All authors declare that there are no conflict of interest in this study.

Appendix A

Table
Table A1. Sequence characteristics of ERF gene family in tea plant.
Table A1. Sequence characteristics of ERF gene family in tea plant.
GeneGene IDLengthMolecular
Weight (D)		Theoretical
pI		Instability
Index		GRAVYAP2 Domain PositionNumber of Transmembrane Domains
CsERF1CSS0000733.121023,215.227.6853.92−0.432106–1700
CsERF2CSS0000882.134738,546.065.9950.24−0.460151–2140
CsERF3CSS0001911.122524,548.559.0361.66−0.59585–1490
CsERF4CSS0002695.124827,194.706.2158.64−0.453114–1730
CsERF5CSS0003530.127730,626.575.8551.37−0.73885–1480
CsERF6CSS0004785.124126,767.789.8657.96−0.72053–1110
CsERF7CSS0005103.141947,136.294.5954.71−0.675108–1540
CsERF8CSS0005145.129633,052.857.8266.02−0.55184–1410
CsERF9CSS0005784.123325,226.119.1863.03−0.58824–840
CsERF10CSS0005931.136741,079.864.4963.44−0.79739–850
CsERF11CSS0007955.115917,417.608.9548.07−0.63060–1180
CsERF12CSS0008086.143046,488.576.4161.60−0.759206–2650
CsERF13CSS0009309.125027,682.076.0256.66−0.506122–1850
CsERF14CSS0009327.121423,759.926.6356.43−0.472110–1740
CsERF15CSS0009615.118320,276.484.9457.36−0.67065–1290
CsERF16CSS0010349.119020,877.154.7364.12−0.51472–1360
CsERF17CSS0010538.114115,375.479.9063.04−0.4829–690
CsERF18CSS0011138.135640,084.824.9557.86−0.675117–1740
CsERF19CSS0012522.121322,962.609.8152.34−0.56819–680
CsERF20CSS0012838.122725,591.868.6946.23−0.75983–1390
CsERF21CSS0013312.125628,777.436.7850.93−0.797122–1790
CsERF22CSS0014363.125628,777.436.7850.93−0.797122–1790
CsERF23CSS0014988.135840,156.724.9555.63−0.711117–1740
CsERF24CSS0015155.123325,236.869.7359.15−0.72836–970
CsERF25CSS0015392.126129,180.905.6351.79−0.598112–1760
CsERF26CSS0016109.128131,339.016.3356.91−0.625129–1880
CsERF27CSS0016950.117619,433.926.2280.30−0.6116–690
CsERF28CSS0017245.118820,599.709.8763.41−0.46236–920
CsERF29CSS0017764.118420,288.839.0768.69−0.5806–690
CsERF30CSS0018119.120523,012.035.7360.26−0.52998–1500
CsERF31CSS0018401.123526,145.829.3952.29−0.82354–1120
CsERF32CSS0020296.122725,118.969.5459.14−0.72931–910
CsERF33CSS0021082.123225,965.955.0460.79−0.525100–1640
CsERF34CSS0021659.138941,806.746.7762.40−0.724166–2250
CsERF35CSS0021688.123326,175.146.0563.54−0.80982–1460
CsERF36CSS0021709.140745,657.784.6060.53−0.91341–980
CsERF37CSS0022692.119020,757.399.9569.35−0.70322–670
CsERF38CSS0022768.123626,410.365.0960.73−0.50898–1560
CsERF39CSS0022828.120122,488.389.7156.35−0.59829–880
CsERF40CSS0023536.125128,008.185.8335.78−0.832129–1920
CsERF41CSS0023891.125628,598.936.0335.61−0.800129–1920
CsERF42CSS0024344.121124,000.845.0743.04−0.73362–1200
CsERF43CSS0024454.139443,916.604.8159.31−0.71344–1010
CsERF44CSS0024711.123126,014.948.3460.37−0.94282–1460
CsERF45CSS0025246.127429,870.659.6751.19−0.58950–1130
CsERF46CSS0026825.115116,983.2110.2441.61−0.7927–690
CsERF47CSS0027366.124827,194.706.2158.64−0.453114–1730
CsERF48CSS0029247.114817,029.635.3674.53−1.18624–880
CsERF49CSS0029398.121724,611.805.5853.91−0.63175–1330
CsERF50CSS0029901.117820,168.028.9351.78−0.6117–700
CsERF51CSS0030082.121123,126.978.7558.05−0.5866–690
CsERF52CSS0030652.118721,042.907.6872.86−0.6376–690
CsERF53CSS0030706.128231,436.126.3356.64−0.629130–1890
CsERF54CSS0030720.118520,465.654.8559.25−0.61765–1290
CsERF55CSS0031448.127130,439.945.3550.70−0.74861–1180
CsERF56CSS0033437.139543,986.654.8059.52−0.71444–1010
CsERF57CSS0034219.122524,994.054.8027.54−0.441112–1670
CsERF58CSS0034413.120923,155.967.7754.72−0.65220–820
CsERF59CSS0034513.120322,908.735.7353.02−0.49573–1370
CsERF60CSS0035247.128532,055.645.2352.65−0.86673–1340
CsERF61CSS0035725.124927,547.016.9357.96−0.559116–1750
CsERF62CSS0036008.118520,465.654.8559.25−0.61765–1290
CsERF63CSS0036725.114315,898.326.8574.17−0.97316–750
CsERF64CSS0037417.122224,644.589.2857.00−0.61529–880
CsERF65CSS0037465.123526,145.829.3952.29−0.82354–1120
CsERF66CSS0037642.142147,271.434.5856.26−0.673108–1540
CsERF67CSS0037694.118820,599.709.8763.41−0.46236–920
CsERF68CSS0038487.123325,236.869.7359.15−0.72836–970
CsERF69CSS0038875.129633,052.857.8266.02−0.55184–1410
CsERF70CSS0039270.123126,116.158.6066.85−0.88582–1460
CsERF71CSS0039495.113715,449.825.5351.69−1.08818–760
CsERF72CSS0039711.118720,886.807.6764.53−0.5776–690
CsERF73CSS0040091.114315,234.309.5746.82−0.2908–650
CsERF74CSS0040214.118720,886.807.6764.53−0.5776–690
CsERF75CSS0040746.118219,782.345.3140.46−0.198105–1580
CsERF76CSS0041141.132536,630.265.9339.75−0.670120–1840
CsERF77CSS0041210.238242,052.384.9639.55−0.713109–1660
CsERF78CSS0041850.114215,811.246.8573.27−0.97516–750
CsERF79CSS0042430.121324,169.085.2940.90−0.70762–1200
CsERF80CSS0042790.122424,175.259.2759.64−0.50740–970
CsERF81CSS0043156.125928,542.915.2059.26−0.503112–1760
CsERF82CSS0043971.119622,341.275.0556.07−0.55250–1070
CsERF83CSS0047280.135638,977.456.2655.96−0.56355–1200
CsERF84CSS0047404.126429,876.829.0755.57−0.802140–2010
CsERF85CSS0047738.124326,935.089.9859.23−0.68753–1110
CsERF86CSS0049535.125027,556.965.7157.01−0.460104–1680
CsERF87CSS0049701.120322,686.024.9372.06−0.66486–1330
CsERF88CSS0049714.142545,620.796.3256.99−0.689209–2600
CsERF89CSS0049793.119722,380.677.6956.65−0.4627–680
CsERF90CSS0049819.140545,060.164.6967.46−0.81547–1030
Table
Table A2. Secondary structure and subcellular location of ERF gene family proteins in tea plant.
Table A2. Secondary structure and subcellular location of ERF gene family proteins in tea plant.
ProteinAlpha Helix (aa)
(%)		Extended Strand (aa)
(%)		Beta Bridge (aa)
(%)		Random Coil (aa)
(%)		Subcellular
Location
CsERF161 (29.05)22 (10.48)0 (0)121 (57.62)Nucleus (9)
CsERF2100 (28.82)38 (10.95)0 (0)190 (54.76)Nucleus
CsERF3103 (31.02)40 (12.05)0 (0)170 (51.20)Nucleus (12)
CsERF463 (25.40)31 (12.50)0 (0)139 (56.05)Nucleus (9)
CsERF576 (27.44)21 (7.58)0 (0)167 (60.29)Nucleus (13)
CsERF663 (26.14)19 (7.88)0 (0)154 (63.90)Nucleus (11.5)
CsERF7117 (27.92)43 (10.26)0 (0)246 (58.71)Nucleus (8.5)
CsERF871 (23.99)33 (11.15)0 (0)173 (58.45)Nucleus (8)
CsERF927 (11.59)43 (18.45)0 (0)155 (66.52)Nucleus (9)
CsERF10160 (43.60)21 (5.72)0 (0)178 (48.50)Nucleus (10.5)
CsERF1145 (28.30)9 (5.66)0 (0)92 (57.86)Nucleus (12)
CsERF12104 (24.19)34 (7.91)0 (0)275 (63.95)Nucleus (14)
CsERF1393 (37.20)23 (9.20)0 (0)128 (51.20)Nucleus (12)
CsERF1457 (26.64)27 (12.62)0 (0)123 (57.48)Nucleus (10)
CsERF1574 (40.44)21 (11.48)0 (0)83 (45.36)Nucleus (12)
CsERF1669 (36.32)25 (13.16)0 (0)89 (46.84)Nucleus (12)
CsERF1728 (19.86)18 (12.77)0 (0)84 (59.57)Nucleus (7)
CsERF1872(20.22)31 (8.71)0 (0)245 (68.82)Nucleus (11)
CsERF1933(15.49)45 (21.13)0 (0)124 (58.22)Nucleus (14)
CsERF2071(31.28)16 (7.05)0 (0)131 (57.71)Nucleus (12)
CsERF2173 (28.52)19 (7.42)0 (0)157 (61.33)Nucleus (7)
CsERF2273 (28.52)19 (7.42)0 (0)157 (61.33)Nucleus (7)
CsERF2373 (20.39)50 (13.97)0 (0)229 (63.97)Nucleus (14)
CsERF2433 (14.16)33 (14.16)0 (0)162 (69.53)Nucleus (13)
CsERF25113 (43.30)20 (7.66)0 (0)120 (45.98)Nucleus (12)
CsERF2665 (23.13)46 (16.37)0 (0)159 (56.58)Nucleus (13)
CsERF2762 (35.23)25 (14.20)0 (0)77 (43.75)Nucleus (7)
CsERF2842 (22.34)31 (16.49)0 (0)107 (56.91)Chloroplast (11)
CsERF2955 (29.89)27 (14.67)0 (0)89 (48.37)Nucleus (10)
CsERF3054 (26.34)21 (10.24)0 (0)118 (57.56)Nucleus (11)
CsERF3146 (19.57)33 (14.04)0 (0)136 (57.87)Nucleus (14)
CsERF3231 (13.66)29 (12.78)0 (0)158 (69.60)Nucleus (9)
CsERF3389 (38.36)21 (9.05)0 (0)108 (46.55)Nucleus (13)
CsERF3496 (24.68)45 (11.57)0 (0)237 (60.93)Nucleus (9.5)
CsERF3594 (40.34)28 (12.02)0 (0)105 (45.06)Nucleus (13)
CsERF36167 (41.03)36 (8.85)0 (0)192 (47.17)Nucleus (14)
CsERF3753 (27.89)39 (20.53)0 (0)83 (43.68)Nucleus (14)
CsERF3894 (39.83)27 (11.44)0 (0)106 (44.92)Nucleus (12)
CsERF3953 (26.37)38 (18.91)0 (0)104 (51.74)Nucleus (9)
CsERF4088 (35.06)12 (4.78)0 (0)148 (58.96)Nucleus (11)
CsERF4184 (32.81)18 (7.03)0 (0)146 (57.03)Nucleus (11)
CsERF4269 (32.70)17 (8.06)0 (0)115 (54.50)Mitochondria (7)
CsERF43165 (41.88)28 (7.11)0 (0)191 (48.48)Nucleus (13)
CsERF4487 (37.66)25 (10.82)0 (0)109 (47.19)Nucleus (14)
CsERF4568 (24.82)35 (12.77)0 (0)158 (57.66)Nucleus (10)
CsERF4662 (41.06)17 (11.26)0 (0)59 (39.07)Mitochondria (7)
CsERF4763 (25.40)31 (12.50)0 (0)139 (56.05)Nucleus (9)
CsERF4861 (41.22)11 (7.43)0 (0)69 (46.62)Nucleus (13)
CsERF4981 (37.33)21 (9.68)0 (0)108 (49.77)Nucleus (13)
CsERF5075 (42.13)9 (5.06)0 (0)88 (49.44)Chloroplast (7)
CsERF5153 (25.12)26 (12.32)0 (0)120 (56.87)Nucleus (9)
CsERF5254 (28.88)22 (11.76)0 (0)102 (54.55)Nucleus (10)
CsERF5364 (22.70)51 (18.09)0 (0)155 (54.96)Nucleus (13)
CsERF5470 (37.84)19 (10.27)0 (0)90 (48.65)Nucleus (12)
CsERF5581 (29.89)21 (7.75)0 (0)158 (58.30)Nucleus (7)
CsERF56170 (43.04)26 (6.58)0 (0)188 (47.59)Nucleus (13)
CsERF5799 (44.00)20 (8.89)0 (0)102 (45.33)Mitochondria (8)
CsERF5839 (18.66)21 (10.05)0 (0)142 (67.94)Nucleus (9)
CsERF5999 (48.77)16 (7.88)0 (0)84 (41.38)Nucleus (10)
CsERF6079 (27.72)29 (10.18)0 (0)163 (57.19)Chloroplast (5)
CsERF6167 (26.91)29 (11.65)0 (0)141 (56.63)Nucleus (10)
CsERF6270 (37.84)19 (10.27)0 (0)90 (48.65)Nucleus (12)
CsERF6345 (31.47)20 (13.99)0 (0)68 (47.55)Nucleus (14)
CsERF6454 (24.32)40 (18.02)0 (0)119 (53.60)Nucleus (10)
CsERF6546 (19.57)33 (14.04)0 (0)136 (57.87)Nucleus (14)
CsERF66129 (30.64)38 (9.03)0 (0)238 (56.53)Nucleus (10)
CsERF6742 (22.34)31 (16.49)0 (0)107 (56.91)Chloroplast (11)
CsERF6833 (14.16)33 (14.16)0 (0)162 (69.53)Nucleus (13)
CsERF6971 (23.99)33 (11.15)0 (0)173 (58.45)Nucleus (8)
CsERF7086 (37.23)26 (11.26)0 (0)114 (49.35)Nucleus (14)
CsERF7152 (37.96)17 (12.41)0 (0)54 (39.42)Nucleus (13)
CsERF7255 (29.41)25 (13.37)0 (0)97 (51.87)Nucleus (7)
CsERF7325 (17.48)21 (14.69)0 (0)83 (58.04)Chloroplast (6)
CsERF7455 (29.41)25 (13.37)0 (0)97 (51.87)Nucleus (7)
CsERF7556 (30.77)20 (10.99)0 (0)99 (54.40)Chloroplast (6)
CsERF7668 (20.92)29 (8.92)0 (0)223 (68.62)Nucleus (13)
CsERF7796 (25.13)32 (8.38)0 (0)244 (63.87)Nucleus (13)
CsERF7842 (29.58)20 (14.08)0 (0)67 (47.18)Nucleus (14)
CsERF7967 (31.46)17 (7.98)0 (0)119 (55.87)Mitochondria (7)
CsERF8044 (19.64)27 (12.05)0 (0)142 (63.39)Chloroplast (7)
CsERF8149 (18.92)47 (18.15)0 (0)147 (56.76)Nucleus (12)
CsERF8278 (39.80)26 (13.27)0 (0)83 (42.35)Nucleus (8)
CsERF8371 (19.94)27 (7.58)0 (0)251 (70.51)Nucleus (14)
CsERF8473 (27.65)14 (5.30)0 (0)166 (62.88)Chloroplast (7)
CsERF8552 (21.40)30 (12.35)0 (0)153 (62.96)Nucleus (9.5)
CsERF8652 (20.80)43 (17.20)0 (0)143 (57.20)Nucleus (11)
CsERF8782 (40.39)20 (9.85)0 (0)91 (44.83)Nucleus (13.5)
CsERF88114 (26.82)34 (8.00)0 (0)261 (61.41)Nucleus (14)
CsERF8978 (39.59)12 (6.09)0 (0)99 (50.25)Mitochondria (6.5)
CsERF90164 (40.49)32 (7.90)0 (0)195 (48.15)Nucleus (11.5)

References

  1. Lu, L.; Chen, H.; Wang, X.; Zhao, Y.; Yao, X.; Xiong, B.; Deng, Y.; Zhao, D. Genome-level diversification of eight ancient tea populations in the Guizhou and Yunnan regions identifies candidate genes for core agronomic traits. Hortic. Res. 2021, 8, 190. [Google Scholar] [CrossRef]
  2. Zhang, W.; Rong, J.; Wei, C.; Gao, L.-M.; Chen, J. Domestication origin and spread of cultivated tea plants. Biodivers. Sci. 2018, 26, 357–372. [Google Scholar] [CrossRef]
  3. Sharangi, A.B. Medicinal and therapeutic potentialities of tea (Camellia sinensis L.)—A review. Food Res. Int. 2009, 42, 529–535. [Google Scholar] [CrossRef]
  4. Yan, Y.; Jeong, S.; Park, C.-E.; Mueller, N.D.; Piao, S.; Park, H.; Joo, J.; Chen, X.; Wang, X.; Liu, J. Effects of extreme temperature on China’s tea production. Environ. Res. Lett. 2021, 16, 044040. [Google Scholar] [CrossRef]
  5. Lata, C.; Prasad, M. Role of DREBs in regulation of abiotic stress responses in plants. J. Exp. Bot. 2011, 62, 4731–4748. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  6. Mitsis, T.; Efthimiadou, A.; Bacopoulou, F.; Vlachakis, D.; Chrousos, G.P.; Eliopoulos, E. Transcription factors and evolution: An integral part of gene expression. World Acad. Sci. J. 2020, 2, 3–8. [Google Scholar] [CrossRef] [Green Version]
  7. Nakano, T.; Suzuki, K.; Fujimura, T.; Shinshi, H. Genome-wide analysis of the ERF gene family in Arabidopsis and rice. Plant Physiol. 2006, 140, 411–432. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  8. Sakuma, Y.; Liu, Q.; Dubouzet, J.G.; Abe, H.; Shinozaki, K.; Yamaguchi-Shinozaki, K. DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs, transcription factors involved in dehydration-and cold-inducible gene expression. Biochem. Biophys. Res. Commun. 2002, 290, 998–1009. [Google Scholar] [CrossRef]
  9. Jofuku, K.D.; Den Boer, B.G.; Van Montagu, M.; Okamuro, J.K. Control of Arabidopsis flower and seed development by the homeotic gene APETALA2. Plant Cell 1994, 6, 1211–1225. [Google Scholar]
  10. Hu, L.; Liu, S. Genome-wide identification and phylogenetic analysis of the ERF gene family in cucumbers. Genet. Mol. Biol. 2011, 34, 624–634. [Google Scholar] [CrossRef] [Green Version]
  11. Zhang, G.; Chen, M.; Chen, X.; Xu, Z.; Guan, S.; Li, L.-C.; Li, A.; Guo, J.; Mao, L.; Ma, Y. Phylogeny, gene structures, and expression patterns of the ERF gene family in soybean (Glycine max L.). J. Exp. Bot. 2008, 59, 4095–4107. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  12. He, S.; Hao, X.; He, S.; Hao, X.; Chen, X. Genome-wide identification, phylogeny and expression analysis of AP2/ERF transcription factors family in sweet potato. BMC Genom. 2021, 22, 748. [Google Scholar] [CrossRef] [PubMed]
  13. Chen, J.; Zhou, Y.; Zhang, Q.; Liu, Q.; Li, L.; Sun, C.; Wang, K.; Wang, Y.; Zhao, M.; Li, H. Structural variation, functional differentiation and expression characteristics of the AP2/ERF gene family and its response to cold stress and methyl jasmonate in Panax ginseng CA Meyer. PLoS ONE 2020, 15, e0226055. [Google Scholar] [CrossRef] [Green Version]
  14. Su, Y.; Ma, Z.; Mao, J.; Li, W.; Cao, X.; Chen, B. Genome-wide identification and characterization of the strawberry (Fragaria vesca) FvAP2/ERF gene family in abiotic stress. Plant Mol. Biol. Report. 2022, 40, 646–660. [Google Scholar] [CrossRef]
  15. Xu, S.; Yao, S.; Huang, R.; Tan, Y.; Huang, D. Transcriptome-wide analysis of the AP2/ERF transcription factor gene family involved in the regulation of gypenoside biosynthesis in Gynostemma pentaphyllum. Plant Physiol. Biochem. 2020, 154, 238–247. [Google Scholar] [CrossRef] [PubMed]
  16. Shigyo, M.; Hasebe, M.; Ito, M. Molecular evolution of the AP2 subfamily. Gene 2006, 366, 256–265. [Google Scholar] [CrossRef]
  17. Magnani, E.; Sjölander, K.; Hake, S. From endonucleases to transcription factors: Evolution of the AP2 DNA binding domain in plants. Plant Cell 2004, 16, 2265–2277. [Google Scholar] [CrossRef] [Green Version]
  18. Balaji, S.; Babu, M.M.; Iyer, L.M.; Aravind, L. Discovery of the principal specific transcription factors of Apicomplexa and their implication for the evolution of the AP2-integrase DNA binding domains. Nucleic Acids Res. 2005, 33, 3994–4006. [Google Scholar] [CrossRef] [Green Version]
  19. Wan, R.; Song, J.; Lv, Z.; Qi, X.; Han, X.; Guo, Q.; Wang, S.; Shi, J.; Jian, Z.; Hu, Q. Genome-wide identification and comprehensive analysis of the AP2/ERF gene family in pomegranate fruit development and postharvest preservation. Genes 2022, 13, 895. [Google Scholar] [CrossRef]
  20. Du, H.; Huang, M.; Zhang, Z.; Cheng, S. Genome-wide analysis of the AP2/ERF gene family in maize waterlogging stress response. Euphytica 2014, 198, 115–126. [Google Scholar] [CrossRef]
  21. Wan, L.; Wu, Y.; Huang, J.; Dai, X.; Lei, Y.; Yan, L.; Jiang, H.; Zhang, J.; Varshney, R.K.; Liao, B. Identification of ERF genes in peanuts and functional analysis of AhERF008 and AhERF019 in abiotic stress response. Funct. Integr. Genom. 2014, 14, 467–477. [Google Scholar] [CrossRef] [PubMed]
  22. Sharma, S.; Chahal, A.; Prasad, H.; Walia, A.; Kumar, R.; Dobhal, S. Identification, phylogeny and transcript profiling of ERF family genes during temperature stress treatment in Pea (Pisum sativum L.). J. Plant Biochem. Biotechnol. 2022, 31, 561–572. [Google Scholar] [CrossRef]
  23. Klay, I.; Gouia, S.; Liu, M.; Mila, I.; Khoudi, H.; Bernadac, A.; Bouzayen, M.; Pirrello, J. Ethylene Response Factors (ERF) are differentially regulated by different abiotic stress types in tomato plants. Plant Sci. 2018, 274, 137–145. [Google Scholar] [CrossRef] [PubMed]
  24. Gao, J.; Zhang, Y.; Li, Z.; Liu, M. Role of ethylene response factors (ERFs) in fruit ripening. Food Qual. Saf. 2020, 4, 15–20. [Google Scholar] [CrossRef] [Green Version]
  25. 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. Plant Physiol. 2010, 152, 1109–1134. [Google Scholar] [CrossRef] [Green Version]
  26. Xing, H.; Jiang, Y.; Zou, Y.; Long, X.; Wu, X.; Ren, Y.; Li, Y.; Li, H.-L. Genome-wide investigation of the AP2/ERF gene family in ginger: Evolution and expression profiling during development and abiotic stresses. BMC Plant Biol. 2021, 21, 561. [Google Scholar] [CrossRef]
  27. Faraji, S.; Filiz, E.; Kazemitabar, S.K.; Vannozzi, A.; Palumbo, F.; Barcaccia, G.; Heidari, P. The AP2/ERF gene family in Triticum durum: Genome-wide identification and expression analysis under drought and salinity stresses. Genes 2020, 11, 1464. [Google Scholar] [CrossRef]
  28. Li, D.; He, Y.; Li, S.; Shi, S.; Li, L.; Liu, Y.; Chen, H. Genome-wide characterization and expression analysis of AP2/ERF genes in eggplant (Solanum melongena L.). Plant Physiol. Biochem. 2021, 167, 492–503. [Google Scholar] [CrossRef]
  29. Oestreich, K.J.; Cobb, R.M.; Pierce, S.; Chen, J.; Ferrier, P.; Oltz, E.M.J.I. Regulation of TCRβ gene assembly by a promoter/enhancer holocomplex. Immunity 2006, 24, 381–391. [Google Scholar] [CrossRef]
  30. Li, P.; Chai, Z.; Lin, P.; Huang, C.; Huang, G.; Xu, L.; Deng, Z.; Zhang, M.; Zhang, Y.; Zhao, X. Genome-wide identification and expression analysis of AP2/ERF transcription factors in sugarcane (Saccharum spontaneum L.). BMC Genom. 2020, 21, 685. [Google Scholar] [CrossRef]
  31. Tang, H.; Bowers, J.E.; Wang, X.; Ming, R.; Alam, M.; Paterson, A.H.J.S. Synteny and collinearity in plant genomes. Science 2008, 320, 486–488. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  32. Yang, Z.; Nielsen, R. Mutation-selection models of codon substitution and their use to estimate selective strengths on codon usage. Mol. Biol. Evol. 2008, 25, 568–579. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  33. Jiao, Y.; Wickett, N.J.; Ayyampalayam, S.; Chanderbali, A.S.; Landherr, L.; Ralph, P.E.; Tomsho, L.P.; Hu, Y.; Liang, H.; Soltis, P.S. Ancestral polyploidy in seed plants and angiosperms. Nature 2011, 473, 97–100. [Google Scholar] [CrossRef] [PubMed]
  34. Zafar, M.M.; Rehman, A.; Razzaq, A.; Parvaiz, A.; Mustafa, G.; Sharif, F.; Mo, H.; Youlu, Y.; Shakeel, A.; Ren, M. Genome-wide characterization and expression analysis of Erf gene family in cotton. BMC Plant Biol. 2022, 22, 134. [Google Scholar] [CrossRef]
  35. Li, D.; Liu, X.; Shu, L.; Zhang, H.; Zhang, S.; Song, Y.; Zhang, Z. Global analysis of the AP2/ERF gene family in rose (Rosa chinensis) genome unveils the role of RcERF099 in Botrytis resistance. BMC Plant Biol. 2020, 20, 533. [Google Scholar] [CrossRef]
  36. Sun, X.; Zhang, L.; Wong, D.C.; Wang, Y.; Zhu, Z.; Xu, G.; Wang, Q.; Li, S.; Liang, Z.; Xin, H.J.T.P.J. The ethylene response factor Va ERF 092 from Amur grape regulates the transcription factor Va WRKY 33, improving cold tolerance. Plant J. 2019, 99, 988–1002. [Google Scholar] [CrossRef]
  37. Zhang, J.; Yin, X.-R.; Li, H.; Xu, M.; Zhang, M.-X.; Li, S.-J.; Liu, X.-F.; Shi, Y.-N.; Grierson, D.; Chen, K.-S. ETHYLENE RESPONSE FACTOR39–MYB8 complex regulates low-temperature-induced lignification of loquat fruit. J. Exp. Bot. 2020, 71, 3172–3184. [Google Scholar] [CrossRef] [PubMed]
  38. Zhang, Z.; Huang, R. Enhanced tolerance to freezing in tobacco and tomato overexpressing transcription factor TERF2/LeERF2 is modulated by ethylene biosynthesis. Plant Mol. Biol. 2010, 73, 241–249. [Google Scholar] [CrossRef]
  39. Heidari, P.; Ahmadizadeh, M.; Izanlo, F.; Nussbaumer, T. In Silico study of the CESA and CSL gene family in Arabidopsis thaliana and Oryza sativa: Focus on post-translation modifications. Plant Gene 2019, 19, 100189. [Google Scholar] [CrossRef]
  40. Xia, E.H.; Li, F.D.; Tong, W.; Li, P.H.; Wu, Q.; Zhao, H.J.; Ge, R.H.; Li, R.P.; Li, Y.Y.; Zhang, Z.Z. Tea Plant Information Archive: A comprehensive genomics and bioinformatics platform for tea plant. Plant Biotechnol. J. 2019, 17, 1938–1953. [Google Scholar] [CrossRef]
  41. Chen, C.; Chen, H.; Zhang, Y.; Thomas, H.R.; Frank, M.H.; He, Y.; Xia, R. TBtools: An integrative toolkit developed for interactive analyses of big biological data. Mol. Plant 2020, 13, 1194–1202. [Google Scholar] [CrossRef]
  42. Miller, D.R.H.; Leek, T.; Schwartz, R.M. A Hidden Markov model information retrieval system. In Proceedings of the 22nd Annual International ACM SIGIR Conference on Research and Development in Information Retrieval, Berkeley, CA, USA, 15–19 August 1999; pp. 214–221. [Google Scholar]
  43. Finn, R.D.; Clements, J.; Eddy, S.R. HMMER web server: Interactive sequence similarity searching. Nucleic Acids Res. 2011, 39 (Suppl. S2), W29–W37. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  44. Marchler-Bauer, A.; Derbyshire, M.K.; Gonzales, N.R.; Lu, S.; Chitsaz, F.; Geer, L.Y.; Geer, R.C.; He, J.; Gwadz, M.; Hurwitz, D.I.; et al. CDD: NCBI’s conserved domain database. Nucleic Acids Res. 2014, 43, D222–D226. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  45. Wu, Z.-J.; Li, X.-H.; Liu, Z.-W.; Li, H.; Wang, Y.-X.; Zhuang, J. Transcriptome-based discovery of AP2/ERF transcription factors related to temperature stress in tea plant (Camellia sinensis). Funct. Integr. Genom. 2015, 15, 741–752. [Google Scholar] [CrossRef]
  46. Gasteiger, E.; Hoogland, C.; Gattiker, A.; Duvaud, S.E.; Wilkins, M.R.; Appel, R.D.; Bairoch, A. Protein Identification and Analysis Tools on the ExPASy Server. In The Proteomics Protocols Handbook; Walker, J.M., Ed.; Humana Press: Totowa, NJ, USA, 2005; pp. 571–607. [Google Scholar]
  47. Krogh, A.; Larsson, B.; von Heijne, G.; Sonnhammer, E.L.L. Predicting transmembrane protein topology with a hidden Markov model: Application to complete genomes. J. Mol. Biol. 2001, 305, 567–580. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  48. Nakai, K.; Horton, P. PSORT: A program for detecting sorting signals in proteins and predicting their subcellular localization. Trends Biochem. Sci. 1999, 24, 34–35. [Google Scholar] [CrossRef]
  49. Chou, K.-C.; Shen, H.-B. Cell-PLoc 2.0: An improved package of web-servers for predicting subcellular localization of proteins in various organisms. Nat. Sci. 2010, 2, 1090. [Google Scholar] [CrossRef] [Green Version]
  50. Geourjon, C.; Deleage, G. SOPMA: Significant improvements in protein secondary structure prediction by consensus prediction from multiple alignments. Bioinformatics 1995, 11, 681–684. [Google Scholar] [CrossRef]
  51. Bailey, T.L.; Boden, M.; Buske, F.A.; Frith, M.; Grant, C.E.; Clementi, L.; Ren, J.; Li, W.W.; Noble, W.S. MEME SUITE: Tools for motif discovery and searching. Nucleic Acids Res. 2009, 37 (Suppl. S2), W202–W208. [Google Scholar] [CrossRef]
  52. Thompson, J.D.; Gibson, T.J.; Higgins, D.G. Multiple sequence alignment using ClustalW and ClustalX. Curr. Protoc. Bioinform. 2003, 2, 2–3. [Google Scholar] [CrossRef]
  53. Kumar, S.; Stecher, G.; Li, M.; Knyaz, C.; Tamura, K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 2018, 35, 1547. [Google Scholar] [CrossRef] [PubMed]
  54. Lescot, M.; Déhais, P.; Thijs, G.; Marchal, K.; Moreau, Y.; Van de Peer, Y.; Rouzé, P.; Rombauts, S. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res. 2002, 30, 325–327. [Google Scholar] [CrossRef] [PubMed]
  55. Zhou, T.; Chen, J.; Huang, Y.; Jin, Z.; Li, J.; Li, Y.; Zeng, X. Genome-Wide Identification and Expression Analysis of the PIN Auxin Transporter Gene Family in Zanthoxylum armatum DC. Agriculture 2022, 12, 1318. [Google Scholar] [CrossRef]
  56. Memelink, J. Regulation of gene expression by jasmonate hormones. Phytochemistry 2009, 70, 1560–1570. [Google Scholar] [CrossRef] [PubMed]
  57. Cheng, C.; An, L.; Li, F.; Ahmad, W.; Aslam, M.; Ul Haq, M.Z.; Yan, Y.; Ahmad, R.M. Wide-range portrayal of AP2/ERF transcription factor family in Maize (Zea mays L.) development and stress responses. Genes 2023, 14, 194. [Google Scholar] [CrossRef]
  58. Jarambasa, T.; Regon, P.; Jyoti, S.Y.; Gupta, D.; Panda, S.K.; Tanti, B. Genome-wide identification and expression analysis of the Pisum sativum (L.) APETALA2/ethylene-responsive factor (AP2/ERF) gene family reveals functions in drought and cold stresses. Genetica 2023, 151, 225–239. [Google Scholar] [CrossRef]
  59. Khaksar, G.; Sirikantaramas, S. Transcriptome-wide identification and expression profiling of the ERF gene family suggest roles as transcriptional activators and repressors of fruit ripening in durian. PLoS ONE 2021, 16, e0252367. [Google Scholar] [CrossRef]
  60. Liu, Y.; Chen, S.; Chen, J.; Wang, J.; Wei, M.; Tian, X.; Chen, L.; Ma, J. Comprehensive analysis and expression profiles of the AP2/ERF gene family during spring bud break in tea plant (Camellia sinensis). BMC Plant Biol. 2023, 23, 206. [Google Scholar] [CrossRef]
  61. Chen, Y.F.; Li, Z.; Su, W.H.; Sun, T.T.; Wang, Z.T.; Cen, G.L.; Que, Y.X.; Meng, Y.T.; Xu, L.P.; Su, Y.C. Identification and expression analysis of AP2/ERF gene family in sugarcane. Chin. J. Appl. Environ. Biol 2022, 28, 67–81. [Google Scholar]
  62. Karanja, B.K.; Xu, L.; Wang, Y.; Tang, M.; Muleke, E.M.M.; Dong, J.; Liu, L. Genome-wide characterization of the AP2/ERF gene family in radish (Raphanus sativus L.): Unveiling evolution and patterns in response to abiotic stresses. Gene 2019, 718, 144048. [Google Scholar] [CrossRef]
  63. Zhu, P.; Chen, Y.; Zhang, J.; Wu, F.; Wang, X.; Pan, T.; Wei, Q.; Hao, Y.; Chen, X.; Jiang, C. Identification, classification, and characterization of AP2/ERF superfamily genes in Masson pine (Pinus massoniana Lamb.). Sci. Rep. 2021, 11, 5441. [Google Scholar] [CrossRef] [PubMed]
  64. Cao, S.; Wang, Y.; Li, X.; Gao, F.; Feng, J.; Zhou, Y. Characterization of the AP2/ERF transcription factor family and expression profiling of DREB subfamily under cold and osmotic stresses in Ammopiptanthus nanus. Plants 2020, 9, 455. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  65. Wang, H.; Ni, D.; Shen, J.; Deng, S.; Xuan, H.; Wang, C.; Xu, J.; Zhou, L.; Guo, N.; Zhao, J. Genome-wide identification of the AP2/ERF gene family and functional analysis of GmAP2/ERF144 for drought tolerance in soybean. Front. Plant Sci. 2022, 13, 848766. [Google Scholar] [CrossRef]
  66. Choi, J.-W.; Choi, H.H.; Park, Y.-S.; Jang, M.-J.; Kim, S. Comparative and expression analyses of AP2/ERF genes reveal copy number expansion and potential functions of ERF genes in Solanaceae. BMC Plant Biol. 2023, 23, 48. [Google Scholar] [CrossRef]
  67. Ji, A.J.; Luo, H.M.; Xu, Z.C.; Zhang, X.; Zhu, Y.J.; Liao, B.S.; Yao, H.; Song, J.Y.; Chen, S.L. Genome-Wide Identification of the AP2/ERF Gene Family Involved in Active Constituent Biosynthesis in Salvia miltiorrhiza. Plant Genome 2016, 9, plantgenome2015-08. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  68. Rao, G.; Sui, J.; Zeng, Y.; He, C.; Zhang, J. Genome-wide analysis of the AP2/ERF gene family in Salix arbutifolia. FEBS Open Bio 2015, 5, 132–137. [Google Scholar] [CrossRef] [Green Version]
  69. Qian, Z.; Rao, X.; Zhang, R.; Gu, S.; Shen, Q.; Wu, H.; Lv, S.; Xie, L.; Li, X.; Wang, X. Genome-Wide Identification, Evolution, and Expression Analyses of AP2/ERF Family Transcription Factors in Erianthus fulvus. Int. J. Mol. Sci. 2023, 24, 7102. [Google Scholar] [CrossRef]
  70. He, Z.; Webster, S.; He, S.Y. Growth–defense trade-offs in plants. Curr. Biol. 2022, 32, R634–R639. [Google Scholar] [CrossRef]
  71. Ren, R.; Wang, H.; Guo, C.; Zhang, N.; Zeng, L.; Chen, Y.; Ma, H.; Qi, J. Widespread whole genome duplications contribute to genome complexity and species diversity in angiosperms. Mol. Plant 2018, 11, 414–428. [Google Scholar] [CrossRef] [Green Version]
  72. Wang, Y.; Chen, F.; Ma, Y.; Zhang, T.; Sun, P.; Lan, M.; Li, F.; Fang, W. An ancient whole-genome duplication event and its contribution to flavor compounds in the tea plant (Camellia sinensis). Hortic. Res. 2021, 8, 176. [Google Scholar] [CrossRef]
  73. Ghorbani, R.; Zakipour, Z.; Alemzadeh, A.; Razi, H. Genome-wide analysis of AP2/ERF transcription factors family in Brassica napus. Physiol. Mol. Biol. Plants 2020, 26, 1463–1476. [Google Scholar] [CrossRef] [PubMed]
  74. Sharma, M.K.; Kumar, R.; Solanke, A.U.; Sharma, R.; Tyagi, A.K.; Sharma, A.K. Identification, phylogeny, and transcript profiling of ERF family genes during development and abiotic stress treatments in tomato. Mol. Genet. Genom. 2010, 284, 455–475. [Google Scholar] [CrossRef] [PubMed]
  75. Ma, Y.; Zhang, F.; Bade, R.; Daxibater, A.; Men, Z.; Hasi, A. Genome-wide identification and phylogenetic analysis of the ERF gene family in melon. J. Plant Growth Regul. 2015, 34, 66–77. [Google Scholar] [CrossRef]
  76. Guo, Z.; He, L.; Sun, X.; Li, C.; Su, J.; Zhou, H.; Liu, X. Genome-Wide Analysis of the Rhododendron AP2/ERF Gene Family: Identification and Expression Profiles in Response to Cold, Salt and Drought Stress. Plants 2023, 12, 994. [Google Scholar] [CrossRef]
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Figure 1. Evolutionary relationship, gene structure, and distribution of conservative motifs of the ERF gene family in tea plant.
Figure 1. Evolutionary relationship, gene structure, and distribution of conservative motifs of the ERF gene family in tea plant.
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Figure 2. Conservative motif of ERF gene family members in tea plant.
Figure 2. Conservative motif of ERF gene family members in tea plant.
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Figure 3. Phylogenetic tree of ERF gene family members in tea plant and Arabidopsis.
Figure 3. Phylogenetic tree of ERF gene family members in tea plant and Arabidopsis.
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Figure 4. Phylogenetic tree of ERF gene family members in tea plant.
Figure 4. Phylogenetic tree of ERF gene family members in tea plant.
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Figure 5. Chromosomal location of ERF gene family members in tea plant. Blue is the tandem repeat gene in CsERFs, and red is the non-tandem repeat gene.
Figure 5. Chromosomal location of ERF gene family members in tea plant. Blue is the tandem repeat gene in CsERFs, and red is the non-tandem repeat gene.
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Figure 6. cis-acting elements of the tea plant ERF gene family.
Figure 6. cis-acting elements of the tea plant ERF gene family.
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Figure 7. Collinearity relationship of ERF gene family members in tea plant.
Figure 7. Collinearity relationship of ERF gene family members in tea plant.
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Figure 8. Expression difference of ERF gene family members in different region of tea plant.
Figure 8. Expression difference of ERF gene family members in different region of tea plant.
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Figure 9. Differences in the expression of ERF gene family members in tea plant under cold stress.
Figure 9. Differences in the expression of ERF gene family members in tea plant under cold stress.
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Table
Table 1. Ka and Ks values of highlighted homologous gene pairs in CsERFs.
Table 1. Ka and Ks values of highlighted homologous gene pairs in CsERFs.
Gene IDKaKsKa/Ks
CsERF43CsERF560.0040.0150.285
CsERF57CsERF410.3670.7850.468
CsERF56CsERF360.1480.7920.187
CsERF43CsERF360.1470.8400.176
CsERF57CsERF220.4721.1630.405
CsERF43CsERF900.3111.9550.159
CsERF76CsERF230.1881.6470.114
CsERF82CsERF380.1350.7530.180
CsERF61CsERF470.1030.6300.164
CsERF2CsERF690.2660.7190.370
CsERF13CsERF590.1310.9120.143
CsERF13CsERF870.3052.6170.116
CsERF71CsERF480.1831.5050.121
CsERF55CsERF420.5592.0830.268
CsERF13CsERF380.2822.2760.124
CsERF55CsERF200.4821.7140.281
CsERF13CsERF330.269NaNNaN
CsERF1CsERF750.3790.8740.434
CsERF89CsERF460.1810.8120.222
CsERF1CsERF300.4751.8020.264
CsERF59CsERF330.2112.6150.081
CsERF46CsERF500.2221.0100.220
CsERF75CsERF300.512NaNNaN
CsERF65CsERF60.1980.7580.261
CsERF31CsERF60.1980.7580.261
CsERF31CsERF690.3231.2840.252
CsERF65CsERF80.3231.2490.259
CsERF41CsERF220.3951.5640.253
CsERF36CsERF100.3191.5550.205
CsERF6CsERF690.4461.5500.287
CsERF84CsERF210.2060.9620.214
CsERF84CsERF220.2060.9620.214
CsERF12CsERF880.1920.5010.383
CsERF42CsERF790.0040.0140.285
CsERF32CsERF800.1180.7060.167
CsERF87CsERF380.3031.8890.161
CsERF87CsERF330.1440.7230.199
CsERF42CsERF200.1460.6690.218
CsERF80CsERF580.2672.3010.116
CsERF9CsERF730.4621.5220.304
CsERF17CsERF730.2360.5330.442
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Zhang, T.; Cai, J.; Wang, S.; Lv, L.; Yuan, D.; Zeng, X.; Li, J.; Li, Y. Identification and Expression Analysis of the Ethylene Response Factor Gene Family in Tea Plant (Camellia sinensis). Agronomy 2023, 13, 1900. https://doi.org/10.3390/agronomy13071900

AMA Style

Zhang T, Cai J, Wang S, Lv L, Yuan D, Zeng X, Li J, Li Y. Identification and Expression Analysis of the Ethylene Response Factor Gene Family in Tea Plant (Camellia sinensis). Agronomy. 2023; 13(7):1900. https://doi.org/10.3390/agronomy13071900

Chicago/Turabian Style

Zhang, Tao, Ju Cai, Sisi Wang, Litang Lv, Dandan Yuan, Xiaofang Zeng, Jianrong Li, and Yan Li. 2023. "Identification and Expression Analysis of the Ethylene Response Factor Gene Family in Tea Plant (Camellia sinensis)" Agronomy 13, no. 7: 1900. https://doi.org/10.3390/agronomy13071900

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