Soybean (Glycine max) expansin gene superfamily origins: segmental and tandem duplication events followed by divergent selection among subfamilies
© Zhu et al.; licensee BioMed Central Ltd. 2014
Received: 30 November 2013
Accepted: 27 March 2014
Published: 11 April 2014
Expansins are plant cell wall loosening proteins that are involved in cell enlargement and a variety of other developmental processes. The expansin superfamily contains four subfamilies; namely, α-expansin (EXPA), β-expansin (EXPB), expansin-like A (EXLA), and expansin-like B (EXLB). Although the genome sequencing of soybeans is complete, our knowledge about the pattern of expansion and evolutionary history of soybean expansin genes remains limited.
A total of 75 expansin genes were identified in the soybean genome, and grouped into four subfamilies based on their phylogenetic relationships. Structural analysis revealed that the expansin genes are conserved in each subfamily, but are divergent among subfamilies. Furthermore, in soybean and Arabidopsis, the expansin gene family has been mainly expanded through tandem and segmental duplications; however, in rice, segmental duplication appears to be the dominant process that generates this superfamily. The transcriptome atlas revealed notable differential expression in either transcript abundance or expression patterns under normal growth conditions. This finding was consistent with the differential distribution of the cis-elements in the promoter region, and indicated wide functional divergence in this superfamily. Moreover, some critical amino acids that contribute to functional divergence and positive selection were detected. Finally, site model and branch-site model analysis of positive selection indicated that the soybean expansin gene superfamily is under strong positive selection, and that divergent selection constraints might have influenced the evolution of the four subfamilies.
This study demonstrated that the soybean expansin gene superfamily has expanded through tandem and segmental duplication. Differential expression indicated wide functional divergence in this superfamily. Furthermore, positive selection analysis revealed that divergent selection constraints might have influenced the evolution of the four subfamilies. In conclusion, the results of this study contribute novel detailed information about the molecular evolution of the expansin gene superfamily in soybean.
Expansins are encoded by a multi-gene family, and are composed of a superfamily of plant cell wall loosening proteins that induce pH-dependent wall extension and stress relaxation in a characteristic and unique manner . Expansins were first identified in studies investigating the mechanism of plant cell wall enlargement, and were isolated from cucumber hypocotyls . Recently, increasing numbers of expansins have been identified in other plant species, including oat , tomato , and maize . According to the nomenclature proposed by Kende et al. , the expansin superfamily in plants may be divided into four subfamilies based on phylogenetic sequence analysis; these subfamilies are designated as α-expansin (EXPA), β-expansin (EXPB), expansin-like A (EXLA), and expansin-like B (EXLB). α-Expansin and β-expansin proteins are known to exhibit cell wall loosening activity, and are involved in cell expansion and other developmental events; however, expansin-like A and expansin-like B are only known from their gene sequences , with no experimental evidence about their activity on the cell wall being published .
Functional studies have shown that expansins are involved in many developmental processes, such as fruit softening , xylem formation , abscission (leaf shedding) , seed germination , and the penetration of pollen tubes [13, 14]. The plant cell wall is composed of cellulose microfibrils, which bind to various glycans, including xyloglucan and xylan. The extension of the cell wall involves the movement and separation of cellulose microfibrils by the process of molecular creeping. α-Expansinis hypothesized to promote such movement, by inducing the local dissociation and slippage of xyloglucans, whereas β-expansin is theorized to work in a similar manner on a different glycan, perhaps xylan . However, no assays have demonstrated that expansins have hydrolytic activity or any other enzymatic activities [15–17].
Expansin proteins are typically 250–275 amino acids long, and contain two domains that are preceded by a signal peptide of 20–30 amino acids in length . Domain I has significant, but distant, homology to glycoside hydrolase family family-45 (GH45) proteins, including a series of conserved cysteines and a His-Phe-Asp (HFD) motif that makes up part of the catalytic site of family-45 endoglucanases [9, 18]. Domain II is distantly related to group-2 grass pollen allergens . Domain II is speculated to be a polysaccharide binding domain based on conserved aromatic and polar residues on the surface of the protein . Only the crystal structure of one bacterial expansin  and the Zea m 1 in maize  have been solved.
The completion of soybean genome sequencing  provides us with an opportunity to improve our understanding about the evolution, and other characteristics, of the expansin superfamily in this plant species. In this study, we identified the expansin genes in the soybean genome, and grouped them into four subfamilies. In addition, the expansion patterns of the expansin gene family in Arabidopsis, rice, and soybean were examined. The results indicated that expansin genes in soybean are generated through tandem and segmental duplication. Analysis of the transcriptome atlas of soybean expansin genes in different tissues under normal conditions indicated notable differential expression among subfamilies. This finding indicates the presence of broad functional divergence in this superfamily. Critical amino acids that are responsible for functional divergence were detected. In addition, the location of the amino acid sites that are responsible for functional divergence and/or positive selection indicated the conservation of domain I and the C terminus. The results presented in this study are expected to facilitate further research on this gene family, and provide new insights about the evolutionary history of expansins.
Genome-wide identification of the expansin gene superfamily in soybean
Sizes of the four expansin subfamilies in different plants
Phylogenetic and structural analysis of expansin genes in soybean
As displayed schematically in Figure 2, 10 types of motif (Additional file 9) were detected. The type, order, and number of motifs were similar in proteins of the same subfamily, but differed to proteins in other subfamilies. In the EXPA subfamily, 85.7% (42 out of 49) of members shared the same eight motif components (motif 1 to 8) in the same order, which was significantly different to that of the other three subfamilies in which the members lacked motifs 3 and 7. Moreover, motif 10 was present in all genes of all subfamilies, except EXPA. Consequently, the motif distribution in EXPA was significantly different to that in the other three subfamilies, leading to the subfamilies EXPB, EXLA, and EXLB having a closer evolutionary and phylogenetic relationship. However, most expansins (77.8%; 7 of 9) in the EXPB subfamily contained motif 2, which was present in all expansins of the EXPA subfamily, but not in the EXLA and EXLB subfamilies. This finding indicates that EXPA and EXPB have a closer evolutionary and phylogenetic relationship compared to EXPA with the EXLA/EXLB subfamilies. Therefore, it indicates that the motif locations of expansins belonging to the same subfamily are conserved, whereas divergence exists among expansins from the four subfamilies.
The exon-intron organization of the expansin genes in soybean was examined by comparing the predicted coding sequences (CDS) with their corresponding genomic sequences through the online software GSDS (http://gsds.cbi.pku.edu.cn/), to obtain more insights about their possible gene structural evolution. Because an ATG sequence is located near to the first initiation codon of GmEXLB10, the software GSDS recognized the subsequent ATG as the initiation codon. Thus, the exon-intron organization of this gene was preceded by a short 5′-UTR, whereas in other genes it was not (Figure 2). Our results showed that genes in the same family generally have similar exon-intron structures, with the same number of exons. For example, all genes from the EXPB and EXLA subfamilies contain four exons, most genes from the EXPA subfamilies contain three exons, while the genes from EXLB families contain five exons. In turn, this finding supported the classification of the expansin genes in soybean. Moreover, this result reflects the divergence in the gene structure of the four subfamilies. In addition, variations are present in the exon-intron structure of genes from the EXPA and EXLB subfamilies, with several genes containing different numbers of exons. Most of the expansin genes in the EXPA subfamily contain three exons, while the remainder contains two or four exons. This variation might have resulted from the loss or gain of exons over a long evolutionary period. Furthermore, comparison of the exon-intron structure among genes from the four subfamilies indicated that the EXPB and EXLA subfamilies are more conserved compared to the EXPA and EXLB subfamilies.
The results of the phylogenetic and structural analysis revealed that each of the four subfamilies was conserved, and that there was also broad diversification among subfamilies. The high degree of sequence identity and similar exon-intron structures of expansin genes within each family indicates that the soybean expansin superfamily has undergone gene duplications throughout evolution. As a result, the expansin gene families contain multiple copies that might partially or completely overlap in function, with the analysis of the soybean gene expansion and expression pattern in this study supporting this hypothesis.
Analysis of expansin gene expansion pattern
Gene duplications are considered to be one of the primary driving forces in the evolution of genomes and genetic systems . Duplicated genes provide raw material for the generation of new genes, which, in turn, facilitate the generation of new functions. Segmental duplication, tandem duplication, and transposition events, such as retroposition and replicative transposition , are considered to represent three principal evolutionary patterns. Of these patterns, segmental and tandem duplications have been suggested to represent two of the main causes of gene family expansion in plants . Segmental duplications multiple genes through polyploidy followed by chromosome rearrangements . It occurs most frequently in plants because most plants are diploidized polyploids and retain numerous duplicated chromosomal blocks within their genomes . Tandem duplications were characterized as multiple members of one family occurring within the same intergenic region or in neighboring intergenic regions . In this study, we defined tandem duplicated genes as adjacent homologous genes on a single chromosome, with no more than one intervening gene. For this analysis, we focused on segmental and tandem duplication events. To gain a greater insight about the expansion pattern of soybean expansin genes in this huge gene family, we identified tandem duplicated clusters based on the gene locus, and searched the Plant Genome Duplication Database  to locate segmentally duplicated pairs. We searched for contiguous expansin genes in both the sharing and neighboring regions. We found that 11 out of 75 genes (14.7%) in this family are tandem repeats in soybean (Figure 1), indicating that tandem duplications have contributed to the expansion of this family. We also tested the hypothesis that segmental duplication events play an important role in the evolution of the expansin superfamily in soybean. We searched each soybean expansin gene in PGDD (http://chibba.agtec.uga.edu/duplication/), and found that 68% (51 of 75) of genes are involved in segmental duplication (Figure 1). Of interest, when we compared the 51 segmentally duplicated genes identified in our study with the results of Du et al. [28, 29], 40 (78.4%; 40 of 51) expansin genes originated from whole genome duplications (WGDs), while the remaining 11 (21.6%; 11 of 51) expansin genes were singletons (GmEXPA2, GmEXPA8, GmEXPA17, GmEXPA21, GmEXPA22, GmEXPA23, GmEXPA29, GmEXPA43, GmEXPA45, GmEXPA47, and GmEXPA49). This finding indicates that the remaining 11 segmentally duplicated expansin genes might be derived from independent duplication events. Therefore, part of the expansin genes in soybean was retained after WGDs. Previous studies have suggested that the genes retained as duplicated pairs after WGD events tend to belong to specific classes, such as transcription factors and members of large multiprotein complexes [30–32], which supports the results of the present study.
Genes involved in tandem duplication and their 4DTv values
Tandem duplicated gene pairs
GmEXPA13 & GmEXPA14
GmEXPB4 & GmEXPB5
GmEXLB4 & GmEXLB5
GmEXLB4 & GmEXLB5
GmEXLB5 & GmEXLB6
GmEXLB11 & GmEXLB12
GmEXLB11 & GmEXLB12
GmEXLB11 & GmEXLB12
GmEXLB12 & GmEXLB13
GmEXLB12 & GmEXLB13
GmEXLB13 & GmEXLB14
Estimates of the dates for the segmental duplication events of expanin gene superfamily in soybean
Number of anchors
(mean ± s.d.)
GmEXPA22 & GmEXPA49
0.100 ± 0.012
GmEXPA8 & GmEXPA47
0.119 ± 0.054
GmEXPA4 & GmEXPA32
0.145 ± 0.024
GmEXPA30 & GmEXPA34
0.149 ± 0.054
GmEXPA24 & GmEXPA27
0.157 ± 0.118
GmEXPA11 & GmEXPA15
0.175 ± 0.141
GmEXPA6 & GmEXPA31
0.177 ± 0.213
GmEXPA9 & GmEXPA13
0.188 ± 0.172
GmEXPA21 & GmEXPA43
0.202 ± 0.166
GmEXPA12 & GmEXPA36
0.205 ± 0.096
GmEXPA2 & GmEXPA23
0.239 ± 0.253
GmEXPA26 & GmEXPA38
0.254 ± 0.219
GmEXPA1 & GmEXPA3
0.270 ± 0.132
GmEXPA18 & GmEXPA28
0.296 ± 0.266
GmEXPA17 & GmEXPA29
0.300 ± 0.179
GmEXPA8 & GmEXPA49
0.453 ± 0.085
GmEXPA16 & GmEXPA35
0.477 ± 0.346
GmEXPA47 & GmEXPA49
0.515 ± 0.139
GmEXPA22 & GmEXPA47
0.531 ± 0.118
GmEXPA8 & GmEXPA22
0.539 ± 0.132
GmEXPA24 & GmEXPA34
0.598 ± 0.189
GmEXPA27 & GmEXPA34
0.613 ± 0.180
GmEXPA24 & GmEXPA30
0.617 ± 0.158
GmEXPA27 & GmEXPA30
0.626 ± 0.155
GmEXPA6 & GmEXPA38
0.633 ± 0.257
GmEXPA2 & GmEXPA36
0.650 ± 0.158
GmEXPA6 & GmEXPA26
0.650 ± 0.177
GmEXPA26 & GmEXPA31
0.680 ± 0.163
GmEXPA23 & GmEXPA36
0.685 ± 0.135
GmEXPA2 & GmEXPA12
0.708 ± 0.099
GmEXPA13 & GmEXPA37
0.710 ± 0.102
GmEXPA9 & GmEXPA37
0.763 ± 0.112
GmEXPA12 & GmEXPA23
0.768 ± 0.078
GmEXPA21 & GmEXPA45
0.790 ± 0.207
GmEXPA43 & GmEXPA45
0.817 ± 0.266
GmEXPB8 & GmEXPB9
0.169 ± 0.077
GmEXPB3 & GmEXPB7
0.397 ± 0.277
GmEXLA1 & GmEXLA2
0.167 ± 0.072
GmEXLB3 & GmEXLB8
0.176 ± 0.117
GmEXLB5 & GmEXLB12
0.191 ± 0.165
GmEXLB7 & GmEXLB15
0.202 ± 0.149
GmEXLB6 & GmEXLB14
0.211 ± 0.179
GmEXLB2 & GmEXLB9
0.236 ± 0.216
GmEXLB2 & GmEXLB15
0.447 ± 0.029
GmEXLB9 & GmEXLB15
0.503 ± 0.068
GmEXLB3 & GmEXLB6
0.513 ± 0.093
GmEXLB3 & GmEXLB12
0.630 ± 0.117
GmEXLB4 & GmEXLB8
0.685 ± 0.227
Overall, these results indicate that the expansin gene superfamily has expanded by both segmental and tandem duplication, particularly segmental duplication. Furthermore, most of the genes involved in segmental duplication were retained after WGDs.
Expression analysis of expansin gene superfamily in soybean
In addition, expansin genes that were clustered in branches in the heatmap exhibited similar transcript abundance profiles. However, most of these genes were not clustered in the phylogenetic tree and were relatively phylogenetically distinct. Only several small phylogenetic clades had largely similar transcript abundance profiles, and were marked on the heatmap in red outlined boxes (Figure 3). Soybean expansins that have high sequence similarity and share expression profiles represent good candidates for the evaluation of gene functions in soybean. Therefore, genes in the red outlined boxes may have a similar function in the same tissues. For example, GmEXPA2 and GmEXPA12, which were clustered in the phylogenetic tree with high sequence similarity only expressed in the root tissue, which indicates that both genes may have the same function in the root tissue.
The transcriptome atlas indicated that all four subfamilies of the soybean expansin superfamily were differentially expressed, which may be associated with the divergence of the promoter regions of the expansin genes. Promoters in the upstream region of genes play key roles in conferring developmental and/or the environmental regulation of gene expression . Thus, profiles of cis-acting elements may provide useful information about the regulatory mechanism of gene expression. A computational tool, PlantCARE , was used to identify cis-acting elements in the 1500-bp DNA sequence upstream of the translation initiation codon of expansin genes in soybean. Four types of cis-acting element were found to be significantly abundant in the promoter region of the soybean expansin gene superfamily (Additional file 12). The first type of cis-acting element enriched in the promoter region is the light-responsive element, which includes the G-box [37, 38], Box 4 , and Box I . The G-box appears to be the most abundant light-responsive element in soybean expansin genes, with a mean number of 1.386 copies, while the G-box is less abundant in EXLB (mean number of 0.8000 copies) compared to the other three subfamilies. Another class of cis-acting elements enriched in the promoter region of expansin genes is the plant hormone-responsive elements, including the TCA-element , TGA-element , and GARE-motif . The salicylic acid-responsive TCA-element appears to be the most abundant hormone-related cis-acting element in soybean expansin genes, indicating that salicylic acid regulates the expression of some soybean expansin genes. The abundance of the TGA-element and GARE-motif in soybean expansin genes indicates that auxin and gibberellin also play roles in regulating soybean expansin gene expression. Other elements are also related to auxin- or gibberellin-responsiveness, such as AuxRR-core , TGA-box , P-box , and TATC-box . These results are consistent with previous studies, which reported that some expansins are regulated by auxin [48, 49] and gibberellin [50, 51]. The third most abundant cis-acting element class contains elements that respond to external environment stresses. We observed that most soybean expansin genes appeared to contain ARE , MBS , HSE , and TC-rich elements . ARE is an element involved in anaerobic induction; hence, we speculated that the anaerobic regulation of expansin expression could be tissue or developmental stage depend. The drought-responsive element MBS is also abundant in the promoter region. With few exceptions, expansin genes contain at least one copy of this element (Additional file 12). These results are consistent with the fact that expansin activities have been found to be influenced by various abiotic stressors, including drought [55, 56] and flooding [57–61]. Circadian elements, which are involved in circadian control , comprise the fourth class of cis-acting element that was abundantly found in the promoter region of soybean expansin genes. PlantCARE analysis showed that soybean expansin genes contain circadian elements, potentially indicating that expansin has a distinct diurnal expression pattern . Promoter analysis demonstrated the presence of a diversity of cis-acting elements in the upstream regions of the soybean expansin gene superfamily. This finding provides further support for the various functional roles of expansins in a wide range of developmental processes related to cell wall modification.
These results indicate that the 75 expansin genes in soybean display differential expression in the four subfamilies, either in the abundance of their transcripts or in their expression patterns under normal growth conditions.
Functional divergence analysis of soybean expansin proteins
Functional divergence between subfamilies of the expansin gene superfamily in soybean
Qk > 0.95
Critical amino acid sites
Qk > 0.95
Critical amino acid sites
θI ± s.e.
θII ± s.e.
0.498 ± 0.079
84C,145 V,172 L
-0.023 ± 0.259
62 T,65 L,103 F,104C,121P,122 M,141G,
143 V,160 F,176 V,177G,190*S,191R,207S
0.783 ± 0.082
45G,54Y,61 N,84C,102 N,104C,
0.136 ± 0.278
18A,45G,54Y,56*Q,60 T,61 N,65 L,67 T,69 L,
161 T,165H,167Y,172*L,176 V,
72 N,75S,76C,82I,102 N,104C,106P,107 N,
181D,184*G,185 V,191R,201 W,
120P,125 F,126D,127 L,133*L,137Q,138Y,
145 V,147Y,154R,155R,160 F,162I,165H,
168 F,170 L,175 N,176 V,180G,181D,185 V,
204 N,205 W,207S,208 N,209 N,210Y,213G
0.572 ± 0.141
-0.081 ± 0.298
54Y,63A,67 T,103 F,106P,121P,125 F,126D,
Furthermore, we predicted that some critical amino acid residues are responsible for functional divergence, with suitable cut-off values being derived from the Qk of each comparison. Given that too many functional divergence-related residues (data not shown) were identified by DIVERGE2 when the empirically Qk value 0.8 was used as a cutoff value, we used Qk > 0.95 to predict CAASs to exclude other sites for further analysis. As a result, a total of 19 CAASs were predicted through type-I functional divergence analysis, whereas 63 amino acid sites with fairly high probability (Qk > 0.95) were identified through type-II functional divergence analysis, which is indicative of a radical shift in evolution rate and amino acid properties to some extent. Furthermore, 12 amino acids are crucial for both the type-I and the type-II functional divergence, indicating that shifts in evolutionary rates and altered amino acid physicochemical properties co-occurred at the these amino acid sites. Hence, these sites probably played important roles in functional divergence during the evolutionary process. In addition, we also noticed that the number of predicted sites (Table 4) within each pair differs between type-I and type-II functional divergence; namely, more CAASs were identified by type-II functional divergence within each subfamily pair. Hence, the functional divergence between the genes of the two groups is mainly attributed to rapid changes in amino acid physiochemical properties, followed by the shift in the evolutionary rate.
Besides, in contrast with EXPA/EXPB and EXPB/EXLB, EXPA/EXLB had relatively larger coefficients of functional divergence (θI & θII) and much more sites that were related to functional divergence. Hence, the functional divergence that exists between EXPA and EXLB is more significant compared with that present in EXPA/EXPB and EXPB/EXLB, although no biological or biochemical function has yet been established for any members of EXLB . In addition, we also deduced that a lesser degree of functional divergence occurred within EXPA/EXPB and EXPB/EXLB based on the coefficients of functional divergence and the number of identified CAASs. Hence, EXPB and EXLB have a much closer phylogenetic relationship compared with EXPA and EXLB, which was also indicated by the motif analysis. The motif analysis showed that the EXPA subfamily has a clearly different motif organization compared to the other two subfamilies, whereas the EXPB and EXLB subfamilies shared similar types and numbers of motifs.
Positive selection analysis
Tests for positive selection among codons of expansin genes using site models
Estimates of parameters
Positively selected sitesb
ω = 0.133
p0 = 0.22607 p1 = 0.55054 p2 = 0.22339
ω1 = 0.02570 ω2 = 0.11359 ω3 = 0.33505
p = 0.99176 q = 5.71801
p0 = 0.99999 p = 0.61117 q = 1.88462
(p1 = 0.00001) ω = 2.02644
Parameters estimation and likelihood ratio tests for the branch-site models
Positive selected sitesa
56Q*,133 L*,166S,169N,186A,172 L*,174 T,190S*,198S,203Q
These results indicate divergent selective constraints on the four subfamilies. The EXPB and EXLA subfamilies are considerably more conserved compared to the EXPA and EXLB subfamilies. Furthermore, the EXPA subfamily might have been subject to the strongest positive selection among the four subfamilies, as the most highly significant positive sites were detected in this subfamily.
Origin of the soybean expansin gene superfamily
Recent research studies have assumed that 70% ~ 80% of angiosperms have undergone duplication events [73–76]. For example, 90% and 62% of Arabidopsis thaliana and Oryza sativa loci have undergone duplication events . As an ancient polyploid, soybean has a highly duplicated genome, with nearly 75% of the genes present occurring in multiple copies . The current investigation revealed the duplication pattern of the soybean expansin gene family. Eleven genes were identified as tandem repeats, indicating that tandem duplication has also contributed to the expansion of the soybean expansin gene superfamily. In addition, 51 genes were found to have evolved from segmental duplication, indicating that segmental duplication probably played a pivotal role in expansin gene expansion in the soybean genome. The genome sequencing results revealed that whole genome duplications (WGD) in soybean occurred at approximately 59 and 13 million years ago (MYA), which is consistent with results of the present study. We inferred that expansion of the expansin gene family occurred along with WGD events, and that these genes were retained during evolution. Previous research has indicated that rapid functional divergence and the biased expression of duplicated genes appear to be major factors promoting their retention in the genome [77–81]. In our study, significant functional divergence was identified among the four subfamilies, with duplicated genes exhibiting diverse expression. For instance, in one duplicated gene pair, GmEXPA30 & GmEXPA34, the two genes were retained after genome duplication events, with only GmEXPA30 being expressed in the leaf, indicating biased expression. Similar cases have also been observed in other segmentally duplicated gene pairs, such as GmEXPA4 & GmEXPA32, GmEXPA6 & GmEXPA31, and GmEXPA18 & GmEXPA28. These results further verified our hypothesis that most of the segmentally duplicated soybean expansin genes have been retained from genome duplication events. Analysis of the expansion pattern of the expansin gene superfamily revealed that the soybean genome had undergone large-scale duplication. Both segmental and tandem duplication are important contributors to the expansion of the expansin gene superfamily.
We also analyzed the expansion pattern of the expansin superfamily in Arabidopsis (Additional file 13) and rice (Additional file 14). The results of the present study showed that 50% (18 of 36) of genes were involved in segmental duplication, while 27.8% (10 of 36) of genes were involved in tandem duplication in Arabidopsis. In comparison, 27.6% (16 of 58) of genes were involved in segmental duplication and 55.2% (32 of 58) of genes were involved in tandem duplication in rice. In soybean, 68% (51 of 75) of genes were involved in segmental duplication and 14.7% (11 of 75) of genes were involved in tandem duplication. Hence, we observed that both segmental and tandem duplication have played significant roles in the expansion of the expansin superfamily in soybean, Arabidopsis, and rice. Previous studies have revealed that genes encoding transcription factors and ribosomal components are significantly over-retained following tetraploidy . However, genes influencing the stress response have an elevated probability of retention following tandem duplication . Expansin genes are associated with cell wall enlargement. However, while these genes are not transcription factors, ribosomal components, or genes that influence stress response, they have expanded through both tandem and segmental duplication, instead of just one form of duplication or the other. More intriguingly, we also noticed that the three species showed species-specific expansion patterns. For instance, segmental duplication seemed to be the predominant form of expansion of the expansin gene superfamilies of the two dicots, Arabidopsis and soybean. In contrast, tandem duplication seemed to be the predominant form of the expansion way for the expansin gene family of the monocot, rice.
The much larger family size of EXPB in rice and EXLB in soybean
Duplication events of the four expansin subfamilies in Soybean, Arabidopsis andrice
67.3% (33 of 49)
44.4% (4 of 9)
100% (2 of 2)
73.3% (11 of 15)
4.1% (2 of 49)
22.2% (2 of 9)
0% (0 of 2)
46.7% (7 of 15)
61.5% (16 of 26)
33.3% (2 of 6)
0% (0 of 3)
0% (0 of 1)
23.1% (6 of 26)
33.3% (2 of 6)
66.7% (2 of 3)
0% (0 of 1)
14.7% (5 of 34)
47.4% (9 of 19)
50% (2 of 4)
0% (0 of 1)
52.9% (18 of 34)
73.7% (14 of 19)
0% (0 of 4)
0% (0 of 1)
Recent research has shown that Zea m 1 (EXPB1 from maize) and orthologous group-1 pollen allergens in other grasses are highly abundant in pollen. These genes may induce extension only in grass cell walls, but are not effective on the walls of dicots, aiding the penetration of the pollen tube through the stigma and style by softening the maternal cell walls [9, 84]. Moreover, β-expansin genes are particularly numerous and abundantly expressed in grasses . In this study, we deduced that the size of the rice EXPB subfamily has increased to adapt to specific functional needs during the long evolutionary timeframe. Alternatively, more genes of the rice EXPB subfamily might have been subjected to a higher degree of post-duplication retention for important functions in rice development. In comparison, the genes of the EXPB subfamily of the other two species might have undergone large-scale gene loss during evolution. The even larger size of the EXLB subfamily in soybean might also reflect adaptations to certain functions or environments. Hence, the EXLB members might have a special function in soybean development; however, experimental evidence has yet to establish their activity in the cell wall .
Functional divergence and positive selection analysis
Gene duplications are considered to be one of the primary driving forces in the evolution of genomes and genetic systems . Typically, an amino acid residue is highly conserved in one duplicate gene, but highly variable in the other one . Amino acid site mutation is frequent, with the accumulation of mutations potentially contributing to the functional divergence of duplicated genes [30, 80, 86, 87]. Through the functional divergence analysis, critical amino acid sites (Table 4) were detected. These sites are major contributors to the functional divergence among the four soybean subfamilies. Rapid functional divergence and the biased expression of duplicated genes is expected to promote retention of the gene of the two homologs, or homoeologs derived from WGD [77–81]. In our study, the expansin gene superfamily has undergone large-scale gene duplication, with many genes being retained after WGD events. Mutations of duplicated genes, and the subsequent selection constraints on them, are expected to lead to functional divergence. At the molecular level, amino acid changes that result in reduced fitness are removed by negative selection, whereas changes that increase fitness are retained by positive selection . Through positive selection analysis, amino acid sites that have undergone strong positive selection (Tables 5, 6) were also identified. Finally, we identified seven sites (56Q, 133 L, 172 L, 184G, 190S, 192 T, and 196P) that were responsible for both functional divergence and positive selection, indicating that these sites were important in the evolutionary history of the expansin gene superfamily in soybean.
Numbers of CAASs for functional divergence and positive selection in specific region of the protein structure
Type-I functional divergence
Type-II functional divergence
Site model of positive selection
Branch-site model of positive selection
Responsible for both functional divergence andpositive selection
No sites responsible for functional divergence and positive selection were found in the C terminus, indicating that the C terminusis stringently conserved. In contrast, six amino acid sites responsible for functional divergence and three amino acid sites responsible for positive selection were found in the N terminus, indicating that this terminus contributes to functional divergence. In addition, the expansins of the N terminus are subject to variation, which might facilitate the adaptiveness of expansins for different functional needs. The N-terminal extension in EXPB1 from maize contained a motif (VPPG-PNITT) that was consistently found, with only minor variation, in group-1 grass pollen allergens, but not in other EXPBs . While the function of this N-terminal extension is unknown, it may contribute to protein recognition, transport, packaging, and the processing of the pollen secretory apparatus .
Previous studies have demonstrated that members of the expansin gene family play important roles in cell enlargement and a variety of other developmental processes. The results of the present study indicate that both tandem and segmental duplication have contributed to the expansion of the expansin gene family in soybean. Species-specific expansion characteristics were identified by comparing the expansion pattern of the expansin gene families in Arabidopsis, soybean, and rice. Segmental duplication seemed to be the predominant form of expansion for the expansin gene superfamilies of the two dicots, Arabidopsis and soybean. In contrast, tandem duplication seemed to be the predominant form of expansion for the expansin gene family of the monocot, rice. Furthermore, positive selection might be the main driving force for the functional divergence of duplicated genes, which might be critical for facilitating plant responses to various stressors throughout their evolutionary history. In addition, divergent selection constraints might have influenced the evolution of the four subfamilies. The results of this study are anticipated to further our understanding about the evolutionary processes of soybean expansin genes, and to help enhance functional genomic studies of expansins in an important model system.
Identification of expansin superfamily genes in soybean
Thirty-five gene sequences of the expansin superfamily in Arabidopsis were collected from EXPANSIN CENTRAL (http://www.personal.psu.edu/fsl/ExpCentral/), and used individually to blast against the soybean genome database in Phytozome v9.1 (http://www.phytozome.net/soybean). Sequences were selected as candidate proteins if their E value was ≤ 1e-10. Finally, the Pfam (http://www.sanger.ac.uk/Software/Pfam/) and the Simple Modular Architecture Research Tool (SMART; http://smart.embl-heidelberg.de/smart/batch.pl) were used to confirm each predicted expansin protein sequence was an expansin superfamily member, sharing domain I (PF03330) and domain II (PF01357). Redundant genes (genes with only one of the two domains, or with unintegrated ORF) were manually removed. Putative genes located on different chromosomes were found for each query sequence. A data file containing all the information from the target genes (including the locations on the chromosomes, genomic sequences, full CDS sequences, protein sequences, and 1500 bp of the nucleotide sequences upstream of the translation initiation codon) were downloaded from the website Phytozome (http://www.phytozome.net). The predicted possible signal peptides were estimated using the SignalP 4.1 server (http://www.cbs.dtu.dk/services/SignalP/). Theoretical pI (isoelectric point) and Mw (molecular weight) values were calculated by ExPASy Compute pI/Mw tool [93–95].
Phylogenetic genetic tree construction and structural analysis
Construction of an unrooted neighbor-joining  phylogenetic tree and bootstrap analysis were conducted using the Molecular Evolutionary Genetics Analysis (MEGA) 5.0 program . Motifs of paralogous expansin proteins were identified statistically using MEME with default settings; however, the maximum number of motifs to find was set at 10. Exon-intron organization of genes from the soybean expansin superfamily were determined by comparing predicted coding sequences (CDS) with their corresponding genomic sequences, using the online software GSDS (http://gsds.cbi.pku.edu.cn/).
Analysis of expansin gene expansion patterns
Soybean expansin genes produced a scattered distribution pattern on chromosomes. In addition, several genes were clearly adjacent to one another based on their loci. Therefore, we focused on the process of segmental and tandem duplication. According to Schauser et al. , an effective way to detect a segmental duplication event is to identify additional paralogous protein pairs in the neighborhood of each family member. Consequently, the synteny blocks of each expansin member were searched in the Plant Genome Duplication Database . Each expansin member was searched in the Plant Genome Duplication Database to identify whether it was involved in segmental duplication. Tandem duplications of the expansin genes in the soybean genome were identified by checking their physical locations on individual chromosomes. Tandem duplicated genes were defined as adjacent homologous genes on a single chromosome, with no more than one intervening gene. For example, Glyma17g15670/Glyma17g15680/Glyma17g15690/Glyma17g15710 were identified as tandem duplicated gene clusters.
Dating the duplication events
The Plant Genome Duplication Database directly provides the Ka and Ks with the corresponding duplicated gene pairs. When dating segmental duplication events, all available anchor points with Ks values between 0 and 1 were used to calculate the average Ks. However, duplicated gene pairs with fewer than three anchor points were deleted. The approximate date of the duplication event was calculated using the mean Ks values from T = Ks/2λ , in which the mean synonymous substitution rate (λ) for Fabaceae is 6.1 × 10-9. For tandem duplication events, the protein sequences of the gene pairs were aligned in Clustal X 1.83, and PAL2NAL  was used to guide the resultant coding sequence (CDS) alignments. Ks, which is the number of synonymous substitutions per site, was determined using the aligned CDS in the Codeml procedure phylogenetic analysis by maximum likelihood (PAML) 4.4  after all alignment gaps were eliminated. 4DTv, which is the transversion rate at four-fold synonymous codon positions, was also calculated by PAML at the same time.
RNA-Seq atlas and promoter analysis
RNA-Seq data were introduced to further analyze the expression of expansin genes, and were obtained from Soybase (http://soybase.org/soyseq/) . The cis-acting elements that regulate gene expression are distributed at 300–3000 bp upstream of the coding region, and sequence restriction was also taken into account in PlantCARE . A total of 1500-bp nucleotide sequences upstream of the coding region for each soybean expansin gene were downloaded from Phytozome, and were submitted to PlantCARE for insilico analysis.
Estimation of functional divergence
The software DIVERGE2 was used to detect the functional divergence between members of the soybean expansin subfamilies . The coefficients of Type-I and Type-II functional divergence, θI and θII, between the soybean expansin subfamilies were calculated. If θI or θII is significantly greater than 0, it means that site-specific altered selective constraints or a radical shift of amino acid physiochemical property occurred after gene duplication and/or speciation . Moreover, a site-specific posterior analysis was used to predict amino acid residues that were crucial for functional divergence. In this analysis, large posterior probability (Qk) indicates a high possibility that the functional constraint (or the evolutionary rate) and/or the radical change in the amino acid property of a site is different between two clusters .
Tests of positive selection
Positive selection was investigated using a maximum likelihood approach by the Codeml procedure in PAML 4.4 , under the site model and branch site model. First, accurate nucleotide sequences and related multiple protein sequence alignments of the soybean expansins were obtained by PAL2NAL . The resulting codon alignments and NJ tree were subsequently used in the Codeml program from the PAML package to calculate the dN/dS (or ω) ratio for each site, and to test different evolutionary models.
In the site model, two pairs of site models in PAML were chosen to test positive selection using the likelihood ratio test (LRT), and to identify positively selected sites in an orthologous group using both naive empirical Bayes (NEB) and Bayes empirical Bayes (BEB) estimation methods. First, models M0 (one ratio) and M3 (discrete) were compared, using a test for heterogeneity between codon sites in the dN/dS ratio value, ω. The second comparison was M7 (beta) vs M8 (beta + ω >1); this comparison is the most stringent test of positive selection . When the LRT indicated positive selection, the BEB method was used to calculate the posterior probabilities that each codon is from the site class of positive selection under models M3 and M8 .
The branch site model assumes that the ω ratio varies between codon sites, and that there are four site classes in the sequence. The first class of sites is highly conserved in all lineages, with a small ω ratio, ω0. The second class includes neutral or weakly constrained sites, for which ω = ω1, where ω1 is near-to or smaller-than 1. In the third and fourth classes, the background lineages show ω0 or ω1, whereas the foreground branches show ω2, which may be greater than 1. When constructing the LRTs, the null hypothesis fixes ω2 = 1, allowing sites to evolve under the negative selection of the background lineages being released from constraint, and to evolve neutrally on the foreground lineage. The alternative hypothesis constrains ω2 ≥ 1 [72, 104]. The posterior probabilities associated with specific codons falling into a site class affected by positive selection were calculated using the BEB method, described by Yang et al. .
Availability of supporting data
The data sets supporting this article are included in:
Additional File 2. Protein sequences data of the expansin gene superfamilies in soybean, Arabidopsis and rice.
Additional File 3. Coding sequences data of the expansin gene superfamily in soybean.
Additional File 4. Genomic sequences data of the expansin gene superfamily in soybean.
Additional File 5. 1500 bp of the nucleotide sequences upstream of the translation initiation codon of the expanisn gene superfamily in soybean.
Additional File 8. The multiple sequence alignment of the soybean expansins.
Additional File 10. The RNA-Seq atlas data of the expansin genes.
Authors would like to thank the National Natural Science Foundation of China (30971783) and the Natural Science Foundation of Beijing, China (5132005) for financial support.
- Cosgrove DJ, Li LC, Cho HT, Hoffmann-Benning S, Moore RC, Blecker D: The growing world of expansins. Plant Cell Physiol. 2002, 43 (12): 1436-1444. 10.1093/pcp/pcf180.PubMedGoogle Scholar
- McQueen-Mason S, Durachko DM, Cosgrove DJ: Two endogenous proteins that induce cell wall extension in plants. Plant Cell Online. 1992, 4 (11): 1425-1433. 10.1105/tpc.4.11.1425.Google Scholar
- Li ZC, Durachko DM, Cosgrove DJ: An oat coleoptile wall protein that induces wall extension in vitro and that is antigenically related to a similar protein from cucumber hypocotyls. Planta. 1993, 191 (3): 349-356.Google Scholar
- Keller E, Cosgrove DJ: Expansins in growing tomato leaves. Plant J. 1995, 8 (6): 795-802. 10.1046/j.1365-313X.1995.8060795.x.PubMedGoogle Scholar
- Wu Y, Sharp RE, Durachko DM, Cosgrove DJ: Growth maintenance of the maize primary root at low water potentials involves increases in cell-wall extension properties, expansin activity, and wall susceptibility to expansins. Plant Physiol. 1996, 111 (3): 765-772.PubMed CentralPubMedGoogle Scholar
- Kende H, Bradford K, Brummell D, Cho HT, Cosgrove DJ, Fleming AJ, Gehring C, Lee Y, McQueen-Mason S, Rose JKC, Voesenek LACJ: Nomenclature for members of the expansin superfamily of genes and proteins. Plant Mol Biol. 2004, 55 (3): 311-314. 10.1007/s11103-004-0158-6.PubMedGoogle Scholar
- Sampedro J, Cosgrove DJ: The expansin superfamily. Genome Biol. 2005, 6: 12-Google Scholar
- Sampedro J, Carey RE, Cosgrove DJ: Genome histories clarify evolution of the expansin superfamily: new insights from the poplar genome and pine ESTs. J Plant Res. 2006, 119 (1): 11-21. 10.1007/s10265-005-0253-z.PubMedGoogle Scholar
- Cosgrove DJ: Loosening of plant cell walls by expansins. Nature. 2000, 407 (6802): 321-326. 10.1038/35030000.PubMedGoogle Scholar
- Gray-Mitsumune M, Mellerowicz EJ, Abe H, Schrader J, Winzéll A, Sterky F, Blomqvist K, McQueen-Mason S, Teeri TT, Sundberg B: Expansins abundant in secondary xylem belong to subgroup A of the α-expansin gene family. Plant Physiol. 2004, 135 (3): 1552-1564. 10.1104/pp.104.039321.PubMed CentralPubMedGoogle Scholar
- Belfield EJ, Ruperti B, Roberts JA, McQueen-Mason S: Changes in expansin activity and gene expression during ethylene-promoted leaflet abscission in Sambucusnigra. J Exp Bot. 2005, 56 (413): 817-823. 10.1093/jxb/eri076.PubMedGoogle Scholar
- Chen F, Bradford KJ: Expression of an expansin is associated with endosperm weakening during tomato seed germination. Plant Physiol. 2000, 124 (3): 1265-1274. 10.1104/pp.124.3.1265.PubMed CentralPubMedGoogle Scholar
- Cosgrove DJ, Bedinger P, Durachko DM: Group I allergens of grass pollen as cell wall-loosening agents. Proc Natl Acad Sci. 1997, 94 (12): 6559-6564. 10.1073/pnas.94.12.6559.PubMed CentralPubMedGoogle Scholar
- Pezzotti M, Feron R, Mariani C: Pollination modulates expression of the PPAL gene, a pistil-specific β-expansin. Plant Mol Biol. 2002, 49 (2): 187-197. 10.1023/A:1014962923278.PubMedGoogle Scholar
- Li LC, Cosgrove DJ: Grass group I pollen allergens (β-expansins) lack proteinase activity and do not cause wall loosening via proteolysis. Eur J Biochem. 2001, 268 (15): 4217-4226. 10.1046/j.1432-1327.2001.02336.x.PubMedGoogle Scholar
- McQueen-Mason SJ, Cosgrove DJ: Expansin mode of action on cell walls (analysis of wall hydrolysis, stress relaxation, and binding). Plant Physiol. 1995, 107 (1): 87-100.PubMed CentralPubMedGoogle Scholar
- McQueen-Mason SJ, Fry SC, Durachko DM, Cosgrove DJ: The relationship between xyloglucanendotransglycosylase and in-vitro cell wall extension in cucumber hypocotyls. Planta. 1993, 190 (3): 327-331.PubMedGoogle Scholar
- Cosgrove DJ: Relaxation in a high-stress environment: the molecular bases of extensible cell walls and cell enlargement. Plant Cell. 1997, 9 (7): 1031-10.1105/tpc.9.7.1031.PubMed CentralPubMedGoogle Scholar
- Kerff F, Amoroso A, Herman R, Sauvagea E, Petrellab S, Filéea P, Charliera P, Jorisa B, Tabuchic A, Nikolaidisc N, Cosgrovec DJ: Crystal structure and activity of Bacillus subtilisYoaJ (EXLX1), a bacterial expansin that promotes root colonization. Proc Natl Acad Sci. 2008, 105 (44): 16876-16881. 10.1073/pnas.0809382105.PubMed CentralPubMedGoogle Scholar
- Yennawar NH, Li LC, Dudzinski DM, Tabuchi A, Cosgrove DJ: Crystal structure and activities of EXPB1 (Zea m 1), a β-expansin and group-1 pollen allergen from maize. Proc Natl Acad Sci. 2006, 103 (40): 14664-14671. 10.1073/pnas.0605979103.PubMed CentralPubMedGoogle Scholar
- Schmutz J, Cannon SB, Schlueter J, Ma J, Mitros T, Nelson W, Hyten DL, Song Q, Thelen JJ, Cheng J, Xu D, Hellsten U, May GD, Yu Y, Sakurai T, Umezawa T, Bhattacharyya MK, Sandhu D, Valliyodan B, Lindquist E, Peto M, Grant D, Shu S, Goodstein D, Barry K, Futrell-Griggs M, Abernathy B, Du J, Tian Z, Zhu L: Genome sequence of the palaeopolyploid soybean. Nature. 2010, 463 (7278): 178-183. 10.1038/nature08670.PubMedGoogle Scholar
- Moore RC, Purugganan MD: The early stages of duplicate gene evolution. Proc Natl Acad Sci. 2003, 100 (26): 15682-15687. 10.1073/pnas.2535513100.PubMed CentralPubMedGoogle Scholar
- Kong H, Landherr LL, Frohlich MW, Leebens-Mack J, Ma H, DePamphilis CW: Patterns of gene duplication in the plant SKP1 gene family in angiosperms: evidence for multiple mechanisms of rapid gene birth. Plant J. 2007, 50 (5): 873-885. 10.1111/j.1365-313X.2007.03097.x.PubMedGoogle Scholar
- Cannon SB, Mitra A, Baumgarten A, Young ND, May G: The roles of segmental and tandem gene duplication in the evolution of large gene families in Arabidopsis thaliana. BMC Plant Biol. 2004, 4 (1): 10-10.1186/1471-2229-4-10.PubMed CentralPubMedGoogle Scholar
- Yu J, Wang J, Lin W, Li S, Li H, Zhou J, Mi P, Dong W, Hu S, Zeng C, Zhang J, Zhang Y, Li R, Xu Z, Li S, Li X, Zheng H, Cong L, Lin L, Yin J, Geng J, Li G, Shi J, Liu J, Lv H, Li J, Wang J, Deng Y, Ran L, Shi X: The genomes of Oryza sativa: a history of duplications. PLoS Biol. 2005, 3 (2): e38-10.1371/journal.pbio.0030038.PubMed CentralPubMedGoogle Scholar
- Ramamoorthy R, Jiang SY, Kumar N, Venkatesh PN, Ramachandran S: A comprehensive transcriptional profiling of the WRKY gene family in rice under various abiotic and phytohormone treatments. Plant Cell Physiol. 2008, 49 (6): 865-879. 10.1093/pcp/pcn061.PubMedGoogle Scholar
- Tang H, Wang X, Bowers JE, Ming R, Alam M, Paterson AH: Unraveling ancient hexaploidy through multiply-aligned angiosperm gene maps. Genome Res. 2008, 18 (12): 1944-1954. 10.1101/gr.080978.108.PubMed CentralPubMedGoogle Scholar
- Du J, Tian Z, Sui Y, Zhao M, Song Q, Cannon SB, Cregan P, Ma J: Pericentromeric effects shape the patterns of divergence, retention, and expression of duplicated genes in the paleopolyploid soybean. Plant Cell Online. 2012, 24 (1): 21-32. 10.1105/tpc.111.092759.Google Scholar
- Yin G, Xu H, Xiao S, Qin Y, Li Y, Yan Y, Hu Y: The large soybean (Glycine max) WRKY TF family expanded by segmental duplication events and subsequent divergent selection among subgroups. BMC Plant Biol. 2013, 13 (1): 148-10.1186/1471-2229-13-148.PubMed CentralPubMedGoogle Scholar
- Blanc G, Wolfe KH: Widespread paleopolyploidy in model plant species inferred from age distributions of duplicate genes. Plant Cell Online. 2004, 16 (7): 1667-1678. 10.1105/tpc.021345.Google Scholar
- Seoighe C, Gehring C: Genome duplication led to highly selective expansion of theArabidopsis thalianaproteome. Trends Genet. 2004, 20 (10): 461-464. 10.1016/j.tig.2004.07.008.PubMedGoogle Scholar
- Maere S, De Bodt S, Raes J, Casneuf T, Van Montagu M, Kuiper M, Van de Peer Y: Modeling gene and genome duplications in eukaryotes. Proc Natl Acad Sci U S A. 2005, 102 (15): 5454-5459. 10.1073/pnas.0501102102.PubMed CentralPubMedGoogle Scholar
- Lee DK, Ahn JH, Song SK, Do Choi Y, Lee JS: Expression of an expansin gene is correlated with root elongation in soybean. Plant Physiol. 2003, 131 (3): 985-997. 10.1104/pp.009902.PubMed CentralPubMedGoogle Scholar
- Libault M, Farmer A, Joshi T, Takahashi K, Langley RJ, Franklin LD, He J, Xu D, May G, Stacey G: An integrated transcriptome atlas of the crop model Glycine max, and its use in comparative analyses in plants. Plant J. 2010, 63 (1): 86-99.PubMedGoogle Scholar
- Xue T, Wang D, Zhang S, Ehlting J, Ni F, Jakab S, Zheng C, Zhong Y: Genome-wide and expression analysis of protein phosphatase 2C in rice and Arabidopsis. BMC Genomics. 2008, 9 (1): 550-10.1186/1471-2164-9-550.PubMed CentralPubMedGoogle Scholar
- 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 (1): 325-327. 10.1093/nar/30.1.325.PubMed CentralPubMedGoogle Scholar
- Sommer H, Saedler H: Structure of the chalcone synthase gene of Antirrhinum majus. Mol Gen Genet MGG. 1986, 202 (3): 429-434. 10.1007/BF00333273.Google Scholar
- Menkens AE, Schindler U, Cashmore AR: The G-box: a ubiquitous regulatory DNA element in plants bound by the GBF family of bZIP proteins. Trends Biochem Sci. 1995, 20 (12): 506-510. 10.1016/S0968-0004(00)89118-5.PubMedGoogle Scholar
- Lois R, Dietrich A, Hahlbrock K, Schulz W: A phenylalanine ammonia-lyase gene from parsley: structure, regulation and identification of elicitor and light responsive cis-acting elements. EMBO J. 1989, 8 (6): 1641-PubMed CentralPubMedGoogle Scholar
- Arguello-Astorga GR, Herrera-Estrella LR: Ancestral multipartite units in light-responsive plant promoters have structural features correlating with specific phototransduction pathways. Plant Physiol. 1996, 112 (3): 1151-1166. 10.1104/pp.112.3.1151.PubMed CentralPubMedGoogle Scholar
- Pastuglia M, Roby D, Dumas C, Cock JM: Rapid induction by wounding and bacterial infection of an S gene family receptor-like kinase gene in Brassica oleracea. Plant Cell Online. 1997, 9 (1): 49-60. 10.1105/tpc.9.1.49.Google Scholar
- Guilfoyle TJ, Hagen G, Li Y, Ulmasov T, Liu ZB, Strabala T, Gee M: Auxin-regulated transcription. Funct Plant Biol. 1993, 20 (5): 489-502.Google Scholar
- Ogawa M, Hanada A, Yamauchi Y, Kuwahara A, Kamiya Y, Yamaguchi S: Gibberellin biosynthesis and response during Arabidopsis seed germination. Plant Cell Online. 2003, 15 (7): 1591-1604. 10.1105/tpc.011650.Google Scholar
- Ballas N, Wong LM, Ke M, Theologis A: Two auxin-responsive domains interact positively to induce expression of the early indoleacetic acid-inducible gene PS-IAA4/5. Proc Natl Acad Sci. 1995, 92 (8): 3483-3487. 10.1073/pnas.92.8.3483.PubMed CentralPubMedGoogle Scholar
- Pascuzzi P, Hamilton D, Bodily K, Arias J: Auxin-induced stress potentiates trans-activation by a conserved plant basic/leucine-zipper factor. J Biol Chem. 1998, 273 (41): 26631-26637. 10.1074/jbc.273.41.26631.PubMedGoogle Scholar
- Kim JK, Cao J, Wu R: Regulation and interaction of multiple protein factors with the proximal promoter regions of a rice high pl α-amylase gene. Mol Gen Genet MGG. 1992, 232 (3): 383-393.PubMedGoogle Scholar
- Jacobsen JV, Gu B: Pp 246-271bler F, Chandler PM. Gibberellin and abscisic acid in germinating cereals. Plant hormones: physiology, biochemistry and molecular biology. Edited by: Davies PJ. 1995, Dordrecht, The Netherlands: Kluwer AcademicGoogle Scholar
- Catalá C, Rose JKC, Bennett AB: Auxin-regulated genes encoding cell wall-modifying proteins are expressed during early tomato fruit growth. Plant Physiol. 2000, 122 (2): 527-534. 10.1104/pp.122.2.527.PubMed CentralPubMedGoogle Scholar
- Hutchison KW, Singer PB, McInnis S, Diaz-Sala C, Greenwood MS: Expansins are conserved in conifers and expressed in hypocotyls in response to exogenous auxin. Plant Physiol. 1999, 120 (3): 827-832. 10.1104/pp.120.3.827.PubMed CentralPubMedGoogle Scholar
- Cho HT, Kende H: Expression of expansin genes is correlated with growth in deepwater rice. Plant Cell Online. 1997, 9 (9): 1661-1671. 10.1105/tpc.9.9.1661.Google Scholar
- Lee Y, Kende H: Expression of β-expansins is correlated with internodal elongation in deepwater rice. Plant Physiol. 2001, 127 (2): 645-654. 10.1104/pp.010345.PubMed CentralPubMedGoogle Scholar
- Klotz KL, Lagrimini LM: Phytohormone control of the tobacco anionic peroxidase promoter. Plant Mol Biol. 1996, 31 (3): 565-573. 10.1007/BF00042229.PubMedGoogle Scholar
- Yamaguchi-Shinozaki K, Shinozaki K: Arabidopsis DNA encoding two desiccation-responsive rd29 genes. Plant Physiol. 1993, 101 (3): 1119-10.1104/pp.101.3.1119.PubMed CentralPubMedGoogle Scholar
- Freitas FZ, Bertolini MC: Genomic organization of the Neurosporacrassagsn gene: possible involvement of the STRE and HSE elements in the modulation of transcription during heat shock. Mol Genet Genomics. 2004, 272 (5): 550-561. 10.1007/s00438-004-1086-5.PubMedGoogle Scholar
- Wu Y, Meeley RB, Cosgrove DJ: Analysis and expression of the α-expansin and β-expansin gene families in maize. Plant Physiol. 2001, 126 (1): 222-232. 10.1104/pp.126.1.222.PubMed CentralPubMedGoogle Scholar
- Jones L, McQueen-Mason S: A role for expansins in dehydration and rehydration of the resurrection plant Craterostigmaplantagineum. FEBS Lett. 2004, 559 (1): 61-65.PubMedGoogle Scholar
- Cho HT, Kende H: Expansins in deepwater rice internodes. Plant Physiol. 1997, 113 (4): 1137-1143. 10.1104/pp.113.4.1137.PubMed CentralPubMedGoogle Scholar
- Huang J, Takano T, Akita S: Expression of α-expansin genes in young seedlings of rice (Oryza sativa L.). Planta. 2000, 211 (4): 467-473. 10.1007/s004250000311.PubMedGoogle Scholar
- Kim JH, Cho HT, Kende H: α-Expansins in the semiaquatic ferns Marsileaquadrifolia and Regnellidiumdiphyllum: evolutionary aspects and physiological role in rachis elongation. Planta. 2000, 212 (1): 85-92. 10.1007/s004250000367.PubMedGoogle Scholar
- Vriezen WH, De Graaf B, Mariani C, Voesenek LA: Submergence induces expansin gene expression in flooding-tolerant Rumexpalustris and not in flooding-intolerant R. acetosa. Planta. 2000, 210 (6): 956-963. 10.1007/s004250050703.PubMedGoogle Scholar
- Colmer TD, Peeters AJM, Wagemaker CAM, Vriezen WH, Ammerlaan A, Voesenek LACJ: Expression of α-expansin genes during root acclimations to O2 deficiency in Rumexpalustris. Plant Mol Biol. 2004, 56 (3): 423-437. 10.1007/s11103-004-3844-5.PubMedGoogle Scholar
- Pichersky E, Bernatzky R, Tanksley SD, Breidenbach RB, Kausch AP, Cashmore AR: Molecular characterization and genetic mapping of two clusters of genes encoding chlorophylla/b-binding proteins inLycopersiconesculentum (tomato). Gene. 1985, 40 (2): 247-258.PubMedGoogle Scholar
- Yamaji N, Ma JF: Spatial distribution and temporal variation of the rice silicon transporter Lsi1. Plant Physiol. 2007, 143 (3): 1306-1313. 10.1104/pp.106.093005.PubMed CentralPubMedGoogle Scholar
- Liu Q, Wang H, Zhang Z, Wu J, Feng Y, Zhu Z: Divergence in function and expression of the NOD26-like intrinsic proteins in plants. BMC Genomics. 2009, 10 (1): 313-10.1186/1471-2164-10-313.PubMed CentralPubMedGoogle Scholar
- Liu Q, Zhu H: Molecular evolution of theMLOgene family in Oryza sativaand their functional divergence. Gene. 2008, 409 (1): 1-10.PubMedGoogle Scholar
- Wang M, Wang Q, Zhao H, Zhang X, Pan Y: Evolutionary selection pressure of forkhead domain and functional divergence. Gene. 2009, 432 (1): 19-25.PubMedGoogle Scholar
- Yang Z: PAML 4: phylogenetic analysis by maximum likelihood. Mol Biol Evol. 2007, 24 (8): 1586-1591. 10.1093/molbev/msm088.PubMedGoogle Scholar
- Yang Z, Nielsen R: Estimating synonymous and nonsynonymous substitution rates under realistic evolutionary models. Mol Biol Evol. 2000, 17 (1): 32-43. 10.1093/oxfordjournals.molbev.a026236.PubMedGoogle Scholar
- Yang Z: PAML: Phylogenetic analysis by maximum likelihood Version 3.14. 2004, London: University College LondonGoogle Scholar
- Anisimova M, Bielawski JP, Yang Z: Accuracy and power of the likelihood ratio test in detecting adaptive molecular evolution. Mol Biol Evol. 2001, 18 (8): 1585-1592. 10.1093/oxfordjournals.molbev.a003945.PubMedGoogle Scholar
- Yang Z: PAML: a program package for phylogenetic analysis by maximum likelihood. Comput Appl Biosci. 1997, 13 (5): 555-556.PubMedGoogle Scholar
- Zhang J, Nielsen R, Yang Z: Evaluation of an improved branch-site likelihood method for detecting positive selection at the molecular level. Mol Biol Evol. 2005, 22 (12): 2472-2479. 10.1093/molbev/msi237.PubMedGoogle Scholar
- Otto SP, Whitton J: Polyploid incidence and evolution. Annu Rev Genet. 2000, 34 (1): 401-437. 10.1146/annurev.genet.34.1.401.PubMedGoogle Scholar
- Blanc G, Hokamp K, Wolfe KH: A recent polyploidy superimposed on older large-scale duplications in the Arabidopsis genome. Genome Res. 2003, 13 (2): 137-144. 10.1101/gr.751803.PubMed CentralPubMedGoogle Scholar
- Bowers JE, Chapman BA, Rong J, Paterson AH: Unravelling angiosperm genome evolution by phylogenetic analysis of chromosomal duplication events. Nature. 2003, 422 (6930): 433-438. 10.1038/nature01521.PubMedGoogle Scholar
- Paterson AH, Bowers JE, Chapman BA: Ancient polyploidization predating divergence of the cereals, and its consequences for comparative genomics. Proc Natl Acad Sci U S A. 2004, 101 (26): 9903-9908. 10.1073/pnas.0307901101.PubMed CentralPubMedGoogle Scholar
- Force A, Lynch M, Pickett FB, Amores A, Yan YL, Postlethwait J: Preservation of duplicate genes by complementary, degenerative mutations. Genetics. 1999, 151 (4): 1531-1545.PubMed CentralPubMedGoogle Scholar
- Lynch M, Force A: The probability of duplicate gene preservation bysubfunctionalization. Genetics. 2000, 154 (1): 459-473.PubMed CentralPubMedGoogle Scholar
- He X, Zhang J: Gene complexity and gene duplicability. Curr Biol. 2005, 15 (11): 1016-1021. 10.1016/j.cub.2005.04.035.PubMedGoogle Scholar
- Sémon M, Wolfe KH: Consequences of genome duplication. Curr Opin Genet Dev. 2007, 17 (6): 505-512. 10.1016/j.gde.2007.09.007.PubMedGoogle Scholar
- Schnable JC, Springer NM, Freeling M: Differentiation of the maize subgenomes by genome dominance and both ancient and ongoing gene loss. Proc Natl Acad Sci. 2011, 108 (10): 4069-4074. 10.1073/pnas.1101368108.PubMed CentralPubMedGoogle Scholar
- Freeling M: The evolutionary position of subfunctionalization, downgraded. 2008Google Scholar
- Hanada K, Zou C, Lehti-Shiu MD, Shinozaki K, Shiu SH: Importance of lineage-specific expansion of plant tandem duplicates in the adaptive response to environmental stimuli. Plant Physiol. 2008, 148 (2): 993-1003. 10.1104/pp.108.122457.PubMed CentralPubMedGoogle Scholar
- Cosgrove DJ: New genes and new biological roles for expansins. Curr Opin Plant Biol. 2000, 3 (1): 73-79. 10.1016/S1369-5266(99)00039-4.PubMedGoogle Scholar
- Zheng Y, Xu D, Gu X: Functional divergence after gene duplication and sequence–structure relationship: a case study of G‒protein alpha subunits. J Exp Zool B Mol Dev Evol. 2007, 308 (1): 85-96.PubMedGoogle Scholar
- Gu X, Zhang Z, Huang W: Rapid evolution of expression and regulatory divergences after yeast gene duplication. Proc Natl Acad Sci U S A. 2005, 102 (3): 707-712. 10.1073/pnas.0409186102.PubMed CentralPubMedGoogle Scholar
- Ha M, Kim ED, Chen ZJ: Duplicate genes increase expression diversity in closely related species and allopolyploids. Proc Natl Acad Sci. 2009, 106 (7): 2295-2300. 10.1073/pnas.0807350106.PubMed CentralPubMedGoogle Scholar
- Fetterman CD, Rannala B, Walter MA: Identification and analysis of evolutionary selection pressures acting at the molecular level in five forkhead subfamilies. BMC Evol Biol. 2008, 8 (1): 261-PubMed CentralPubMedGoogle Scholar
- Arnold K, Bordoli L, Kopp J, Schwede T: The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics. 2006, 22 (2): 195-201. 10.1093/bioinformatics/bti770.PubMedGoogle Scholar
- Kiefer F, Arnold K, Künzli M, Bordoli L, Schwede T: The SWISS-MODEL Repository and associated resources. Nucleic Acids Res. 2009, 37 (suppl 1): D387-D392.PubMed CentralPubMedGoogle Scholar
- Peitsch M: Protein modeling by E-mail. Biotechnology. 1995, 13: 658-660. 10.1038/nbt0795-658.Google Scholar
- Li LC, Bedinger PA, Volk C, Jones AD, Cosgrove DJ: Purification and characterization of four β-expansins (Zea m 1 isoforms) from maize pollen. Plant Physiol. 2003, 132 (4): 2073-2085. 10.1104/pp.103.020024.PubMed CentralPubMedGoogle Scholar
- Bjellqvist B, Hughes GJ, Pasquali C, Paquet N, Ravier F, Sanchez JC, Frutiger S, Hochstrasser D: The focusing positions of polypeptides in immobilized pH gradients can be predicted from their amino acid sequences. Electrophoresis. 1993, 14 (1): 1023-1031. 10.1002/elps.11501401163.PubMedGoogle Scholar
- Bjellqvist B, Basse B, Olsen E, Celis JE: Reference points for comparisons of two‒dimensional maps of proteins from different human cell types defined in a pH scale where isoelectric points correlate with polypeptide compositions. Electrophoresis. 1994, 15 (1): 529-539. 10.1002/elps.1150150171.PubMedGoogle Scholar
- Gasteiger E, Hoogland C, Gattiker A, Wikins MR, Appel RD, Bairoch A: Protein identification and analysis tools on the ExPASy server[M]//The proteomics protocols handbook. 2005, USA: Humana PressGoogle Scholar
- Saitou N, Nei M: The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987, 4 (4): 406-425.PubMedGoogle Scholar
- Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S: MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011, 28 (10): 2731-2739. 10.1093/molbev/msr121.PubMed CentralPubMedGoogle Scholar
- Schauser L, Wieloch W, Stougaard J: Evolution of NIN-like proteins in Arabidopsis, rice, and Lotus japonicus. J Mol Evol. 2005, 60 (2): 229-237. 10.1007/s00239-004-0144-2.PubMedGoogle Scholar
- Nei M, Kumar S: Molecular evolution and phylogenetics. 2000, Oxford: Oxford University PressGoogle Scholar
- Lynch M, Conery JS: The evolutionary fate and consequences of duplicate genes. Science. 2000, 290 (5494): 1151-1155. 10.1126/science.290.5494.1151.PubMedGoogle Scholar
- Suyama M, Torrents D, Bork P: PAL2NAL: robust conversion of protein sequence alignments into the corresponding codon alignments. Nucleic Acids Res. 2006, 34 (suppl 2): W609-W612.PubMed CentralPubMedGoogle Scholar
- Severin AJ, Woody JL, Bolon YT, Joseph B, Diers BW, Farmer AD, Muehlbauer GJ, Nelson RT, Grant D, Specht JE, Graham MA, Cannon SB, May GD, Vance CP, Shoemaker RC: RNA-Seq Atlas of Glycine max: a guide to the soybean transcriptome. BMC Plant Biol. 2010, 10 (1): 160-10.1186/1471-2229-10-160.PubMed CentralPubMedGoogle Scholar
- Gu X: A simple statistical method for estimating type-II (cluster-specific) functional divergence ofprotein sequences. Mol Biol Evol. 2006, 23 (10): 1937-1945. 10.1093/molbev/msl056.PubMedGoogle Scholar
- Yang Z, Nielsen R: Codon-substitution models for detecting molecular adaptation at individual sites along specific lineages. Mol Biol Evol. 2002, 19 (6): 908-917. 10.1093/oxfordjournals.molbev.a004148.PubMedGoogle Scholar
- Yang Z, Wong WSW, Nielsen R: Bayes empirical Bayes inference of amino acid sites under positive selection. Mol Biol Evol. 2005, 22 (4): 1107-1118. 10.1093/molbev/msi097.PubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.