Growth-Regulating Factor Gene Family Research Articles

Genome-wide identification and expression analysis of growth-regulating factors in Dendrobium officinale and Dendrobium chrysotoxum.

Dendrobium, one of the largest genera in Orchidaceae, is popular not only for its aesthetic appeal but for its significant medicinal value. Growth-regulating factors (GRFs) play an essential role in plant growth and development. However, there is still a lack of information about the evolution and biological function analysis of the GRF gene family among Dendrobiumspecies. Growth-regulating factors from Dendrobium officinale Kimura et Migo and Dendrobium chrysotoxum Lindl. were identified by HMMER and BLAST. Detailed bioinformatics analysis was conducted to explore the evolution and function of GRF gene family in D. officinale and D. chrysotoxum using genomic data, transcriptome data and qRT-PCR technology. Here, we evaluated the evolution of the GRF gene family based on the genome sequences of D. officinale and D. chrysotoxum. Inferred from phylogenetic trees, the GRF genes were classified into two clades, and each clade contains three subclades. Sequence comparison analysis revealed relatively conserved gene structures and motifs among members of the same subfamily, indicating a conserved evolution of GRF genes within Dendrobiumspecies. However, considering the distribution of orthologous DoGRFs and DcGRFs, and the differences in the number of GRFs among species, we suggest that the GRF gene family has undergone different evolutionary processes. A total of 361 cis-elements were detected, with 33, 141, and 187 related to plant growth and development, stress, and hormones, respectively. The tissue-specific expression of GRFs showed that DoGRF8 may have a significant function in the stem elongation of D. officinale. Moreover, four genes were up-regulated under Methyl-jasmonic acid/methyl jasmonate (MeJA) treatment, showing that DoGRFs and DcGRFs play a crucial role in stress response. These findings provide valuable information for further investigations into the evolution and function of GRF genes in D. officinale and D. chrysotoxum.

Identification, Evolution and Expression Analysis of GRF Family Reveals Their Involvement in Shoot Growth and Abiotic Stress Response in Moso Bamboo

Growth-regulating factors (GRFs) play an important role in regulating plant organ development, acting primarily as positive regulators of cell proliferation. However, research on the evolutionary history and expression patterns of the moso bamboo GRF family has been limited. In this study, a total of 24 GRFs have been identified in the Moso bamboo genome, and they have been categorized into four subfamilies. Estimation of the divergence time of paralogous gene pairs provided evidence supporting the significant contribution of recent whole-genome duplication events in the expansion of the GRF gene family. Sliding window analysis revealed that coding regions of a few PheGRFs, including the WRC and QLQ domains, may have undergone positive selection, possibly due to the redundant functions of paralogous genes. Coexpression network analysis further revealed the regulatory role of miR396 and various lncRNAs in controlling PheGRF expression. Based on the analysis of tissue-specific expression patterns using transcriptome sequencing, qRT-PCR results, and in situ hybridization, it was observed that most GRFs, particularly PheGRF6a and PheGRF9b, exhibited high levels of accumulation in the moso bamboo shoot. This suggests that the involvement of most PheGRF genes may be crucial for the growth and development of the bamboo shoot. A yeast two-hybrid screening revealed interactions between PheGRF9b and several proteins associated with plant growth and development, including PH02Gene11097.t1 (GIF3), PH02Gene37618.t (Phytochrome B), and PH02Gene01921.t3 (WD40). Based on transcriptome expression analysis, it was observed that a substantial number of PheGRFs exhibited significant variations under cold or drought stress treatments, and most of these genes were found to be downregulated, suggesting their role as abiotic stress-responsive genes. Our findings offer new insights into the GRF family of moso bamboo and provide some experimental evidence to support further gene functional validation research of PheGRF.

Genome-Wide Identification and Characterization of the GRF Gene Family in Melastoma dodecandrum.

Growth-regulating factor (GRF) is a kind of transcription factor unique to plants, playing an important role in the flowering regulation, growth, and development of plants. Melastoma dodecandrum is an important member of Melastomataceae, with ornamental, medicinal, and edible benefits. The identification of the GRF gene family in M. dodecandrum can help to improve their character of flavor and continuous flowering. The members of the GRF gene family were identified from the M. dodecandrum genome, and their bioinformatics, selective pressure, and expression patterns were analyzed. The results showed that there were 20 GRF genes in M. dodecandrum. Phylogenetic analysis showed that the 71 GRF genes from M. dodecandrum, Arabidopsis thaliana, Camellia sinensis, and Oryza sativa can be divided into three clades and six subclades. The 20 GRF genes of M. dodecandrum were distributed in twelve chromosomes and one contig. Furthermore, the gene structure and motif analysis showed that the intron and motif within each clade were very similar, but there were great differences among different clades. The promoter contained cis-acting elements related to hormone induction, stress, and growth and development. Different transcriptomic expression of MdGRFs indicated that MdGRFs may be involved in regulating the growth and development of M. dodecandrum. The results laid a foundation for further study on the function and molecular mechanism of the M. dodecandrum GRF gene family.

Genome-wide identification of GRF gene family and their contribution to abiotic stress response in pitaya (Hylocereus polyrhizus)

Growth-regulating factors (GRFs) are plant-specific transcription factors identified in many land plants. Recently, their indispensable roles in stress response are highlighted. In present work, 11 HpGRFs were cloned in pitaya. Segmental duplication is considered essential for the expansion of HpGRFs. A phylogenetic tree suggested that GRFs could be divided into eight categories, among which G-I was a Caryophyllales-specific one. The categorization was further evidenced by differences in the gene structure, collinearity, protein domain of HpGRFs. Five miR396 hairpins giving rise to two types of matured miR396s were identified in pitaya via sRNA-Seq in combination with bioinformatic analysis. Parallel analysis of RNA ends proved that HpGRFs except HpGRF5 were degraded by miR396-directed cleavages at the regions which code the conserved WRC motifs of HpGRFs. Multiple cis-regulatory elements were discovered in the promoters of HpGRFs. Among the elements, most are involved in stress and phytohormone response as well as plant growth, indicating a crosstalk between them. Expression analysis showed the responsive patterns of the miR396-GRF module under abiotic stresses. To conclude, our work systematically identified the miR396-targeted HpGRFs in pitaya and confirmed their involvement in stress response, providing novel insights into the comprehensive understanding of the stress resistance of pitaya.

Genome-Wide Analysis of the Growth-Regulating Factor (GRF) Family in Aquatic Plants and Their Roles in the ABA-Induced Turion Formation of Spirodela polyrhiza.

Growth-regulating factors (GRFs) are plant-specific transcription factors that play essential roles in regulating plant growth and stress response. The GRF gene families have been described in several terrestrial plants, but a comprehensive analysis of these genes in diverse aquatic species has not been reported yet. In this study, we identified 130 GRF genes in 13 aquatic plants, including floating plants (Azolla filiculoides, Wolffia australiana, Lemna minuta, Spirodela intermedia, and Spirodela polyrhiza), floating-leaved plants (Nymphaea colorata and Euryale ferox), submersed plants (Zostera marina, Ceratophyllum demersum, Aldrovanda vesiculosa, and Utricularia gibba), an emergent plant (Nelumbo nucifera), and an amphibious plant (Cladopus chinensis). The gene structures, motifs, and cis-acting regulatory elements of these genes were analyzed. Phylogenetic analysis divided these GRFs into five clusters, and ABRE cis-elements were highly enriched in the promoter region of the GRFs in floating plants. We found that abscisic acid (ABA) is efficient at inducing the turion of Spirodela polyrhiza (giant duckweed), accompanied by the fluctuated expression of SpGRF genes in their fronds. Our results provide information about the GRF gene family in aquatic species and lay the foundation for future studies on the functions of these genes.

Comprehensive Analysis for GRF Transcription Factors in Sacred Lotus (Nelumbo nucifera).

Sacred lotus (Nelumbo nucifera) is an aquatic perennial plant with essential food, ornamental, and pharmacological value. Growth-regulating factor (GRF) is a transcription factor (TF) family that plays an important role in regulating the growth and development of plants. In this study, a comprehensive analysis of the GRF family in N. nucifera was performed, and its role in N. nucifera development was studied. A total of eight GRF genes were identified in the N. nucifera genome. Phylogenetic analysis divided the 38 GRF genes into six clades, while the NuGRFs only contained five clades. The analyses of gene structures, motifs, and cis-acting regulatory elements of the GRF gene family were performed. In addition, the chromosome location and collinearity were analyzed. The expression pattern based on transcriptomic data and real-time reverse transcription-quantitative PCR (qRT-PCR) revealed that the GRF genes were expressed in multiple organs and were abundant in actively growing tissues, and the expression levels decreased as the age of N. nucifera increased. Then, 3D structures of the NuGRF proteins were predicted by homology modeling. Finally, the subcellular localization of GRF1 was ascertained in the tobacco leaf through a vector. Therefore, this study provides a comprehensive overview of the GRF TF family in N. nucifera.

Genome-wide analysis of the GRF gene family and their expression profiling in peach (Prunus persica)

ABSTRACT Growth-regulating factors (GRFs) are plant-specific transcription factors with vital roles in multiple biological processes. Although GRFs have been identified in various plant species, a comprehensive analysis of GRF genes in peach (Prunus persica) has not yet been reported. In this study, 10 PpGRF genes distributed on 6 chromosomes were identified in peach genome and their properties were analyzed systematically. Expression pattern analysis suggested that most PpGRFs were preferentially expressed in young tissues. Multiple types of cis-elements were observed in PpGRF promoters, and PpGRFs positively respond to ultraviolet B-rays (UVB) and gibberellin (GA) treatments at the transcriptional level. Also, the content of gibberellic acid 3 (GA3) and indole-3-acetic acid (IAA) changed significantly after UVB irradiation. PpGRF 3, 4, 5, 6, 7, 9 and 10 positive responses to UVB and GA3 signals. The evolutionary patterns and expression profiles of PpGRFs detected in this study increase understanding of the important roles in peach.

Identification, Molecular Characteristics, and Evolution of GRF Gene Family in Foxtail Millet (Setaria italica L.).

Growth-regulating factor (GRF) is a multigene family that plays a vital role in the growth and development of plants. In the past, the GRF family of many plants has been studied. However, there is not a report about identification and evolution of GRF in foxtail millet (Setaria italia). Here, we identified 10 GRF genes in foxtail millet. Seven (70.00%) were regulated by Sit-miR396, and there were 19 optimal codons in GRFs of foxtail millet. Additionally, we found that WGD or segmental duplication have affected GRFs in foxtail millet between 15.07 and 45.97 million years ago. Regarding the GRF gene family of land plants, we identified a total of 157 GRF genes in 15 representative land plants. We found that GRF gene family originated from Group E, and the GRF gene family in monocots was gradually shrinking. Also, more loss resulted from the small number of GRF genes in lower plants. Exploring the evolution of GRF and functional analysis in the foxtail millet help us to understand GRF better and make a further study about the mechanism of GRF. These results provide a basis for the genetic improvement of foxtail millet and indicate an improvement of the yield.

Genome-wide identification and expression analysis of the growth regulating factor (GRF) family in Jatropha curcas.

GRF genes have been confirmed to have important regulatory functions in plant growth, development and response to abiotic stress. Although the genome of Jatropha curcas is sequenced, knowledge about the identification of the species' GRF genes and their expression patterns is still lacking. In this study, we characterized the 10 JcGRF genes. A detailed investigation into the physic nut GRF gene family is performed, including analysis of the exon-intron structure, conserved domains, conserved motifs, phylogeny, chromosomal locations, potential small RNA targets and expression profiles under both normal growth and abiotic stress conditions. Phylogenetic analysis indicated that the 10 JcGRF genes were classified into five groups corresponding to group I, II, III, IV and V. The analysis of conserved domains showed that the motifs of JcGRF genes were highly conserved in Jatropha curcas. Expression analysis based on RNA-seq and qRT-PCR showed that almost all JcGRF genes had the highest expression in seeds, but very low expression was detected in the non-seed tissues tested, and four JcGRF genes responded to at least one abiotic stress at at least one treatment point. Our research will provide an important scientific basis for further research on the potential functions of JcGRF genes in Jatropha curcas growth and development, and response to abiotic stress, and will eventually provide candidate genes for the breeding of Jatropha curcas.

Genome-Wide Identification and Analysis of the Growth-Regulating Factor (GRF) Gene Family and GRF-Interacting Factor Family in Triticum aestivum L.

Growth-regulating factors (GRFs) are unique transcription factors in plants. GRFs can interact with SNH (SYT N-terminal homology) domains in GRF-interacting factor (GIF) proteins via the N-terminal QLQ (Gln, Leu, Gln) domain to form functional complexes and participate in the regulation of downstream gene expression. In this study, we systematically identified the GRF gene family and GIF gene family in wheat and its relatives comprising Triticum urartu, Triticum dicoccoides, and Aegilops tauschii. Thirty GRF gene members are present in wheat, which are distributed on 12 chromosomes and they have 2-5 protein-coding regions. They all contain QLQ and WRC (Trp, Arg, Cys) conserved domains. Wheat possesses only eight members of the GIF gene family, which are distributed on six chromosomes. All wheat GIF (TaGIF) proteins have highly conserved SNH and QG (Gln, Gly) domains. The wheat GRF (TaGRF) gene family has 13 pairs of segmental duplication genes and no tandem duplication genes; the TaGIF gene family has two pairs of segmental duplication genes and no tandem duplication genes. It is speculated that segmental duplication events may be the main reason for the amplification of TaGRF gene family and TaGIF gene family. Based on published transcriptome data and qRT-PCR results of 8 TaGRF genes and 4 TaGIF genes, all of the genes responded strongly to osmotic stress, and the expression levels of TaGRF21 and TaGIF5 were also significantly upregulated under drought and cold stress conditions. The results obtained in this study may facilitate further investigations of the functions of TaGRF genes and TaGIF genes in order to identify candidate genes for use in stress-resistant wheat breeding programs.

Biological roles and an evolutionary sketch of the GRF-GIF transcriptional complex in plants

GROWTH-REGULATING FACTORs (GRFs) are sequence-specific DNA-binding transcription factors that regulate various aspects of plant growth and development. GRF proteins interact with a transcription cofactor, GRF-INTERACTING FACTOR (GIF), to form a functional transcriptional complex. For its activities, the GRF-GIF duo requires the SWITCH2/SUCROSE NONFERMENTING2 chromatin remodeling complex. One of the most conspicuous roles of the duo is conferring the meristematic potential on the proliferative and formative cells during organogenesis. GRF expression is post-transcriptionally down-regulated by microRNA396 (miR396), thus constructing the GRF-GIF-miR396 module and fine-tuning the duo’s action. Since the last comprehensive review articles were published over three years ago, many studies have added further insight into its action and elucidated new biological roles. The current review highlights recent advances in our understanding of how the GRF-GIF-miR396 module regulates plant growth and development. In addition, I revise the previous view on the evolutionary origin of the GRF gene family.

Genome-wide identification and expression analysis of GRF genes regulating apple tree architecture

Growth-regulating factor (GRF) proteins comprise a family of plant-specific transcription factors that play crucial roles in regulating plant architecture. However, information pertaining to the GRF gene family in apple (Malus × domestica) is limited. In the present study, 16 GRF family members were identified in the apple genome, and nine were predicted to be putative targets of apple microRNA396 (miR396). Phylogenetic and synteny analyses of MdGRF genes indicated that gene duplication events have likely contributed to the expansion and evolution of MdGRFs. Moreover, several pairs of MdGRF/AtGRF homologs were identified. The tissue-specific expression patterns of MdmiR396–MdGRF genes in different tissues and in the ArrayExpress database were subsequently characterized. The analyses indicated that they had opposing functions. Exogenous brassinolide, gibberellic acid, 1-naphthylacetic acid, and 6-benzylaminopurine increased primary shoot lengths and diameters, and elevated the expression levels of several MdGRF genes. The expression levels of MdmiR396 and MdGRF genes were reduced in the standard cultivar “Nagafu 2” compared with its short branch spur-type bud mutation “Yanfu 6,” and in the standard graft combination Nagafu 2/Nagafu 2 compared with the dwarfing graft combination Nagafu 2/“Malling 9.” Thus, MdmiR396-MdGRF may be involved in the regulation of apple tree size in response to hormone signals and in different apple varieties and grafting combinations. This work provides a basis for further functional research on MdmiR396-MdGRFs and their potential involvement in the regulation of apple tree architecture.

Characterization and expression analysis of growth regulating factor (GRF) family genes in cucumber

The growth regulating factor (GRF) family is a conserved class of transcription factors involved in various biological processes in plants. However, there have been only a few studies of the GRF family genes in cucumber, Cucumis sativus (Cs). In this study, we identified and characterized 8 CsGRF genes in cucumber. Two highly conserved domains, QLQ and WRC, were identified to be present in all CsGRF proteins. In addition, three less conserved domains (FFD, TQL, and GGPL) were also detected in some CsGRF members. Based on phylogenetic analysis, the GRF genes from cucumber, Arabidopsis, tomato, rice and maize could be classified into 10 groups, and CsGRFs were clustered closer with the GRF genes from dicots (Arabidopsis and tomato) than with those from monocots (rice and maize). Promoter analysis revealed that the CsGRF genes were involved in cucumber growth and development as well as in responses to various hormones and stresses. Transcriptome data showed that the CsGRF genes have distinct expression patterns in different tissues, especially in ovaries and leaves. Expression profiling analysis indicated that all CsGRF genes were responsive to salt and drought stress treatments. These results demonstrate that the cucumber GRF gene family may function in organ development and plant stress responses.

Genome-wide identification and analysis of the growth-regulating factor family in tobacco (Nicotiana tabacum)

Growth-regulating factors (GRFs) are plant-specific transcription factors that have important functions in regulating plant growth and development. GRF gene families have been described in several plant species, but a comprehensive analysis of the GRF gene family in tobacco has not yet been reported. In this study, we identified 25 NtabGRF genes in N. tabacum. The gene structures, motifs, and cis-acting regulatory elements of the NtabGRF genes were analyzed. Phylogenetic analysis divided the genes into six clusters. Additionally, highly conserved regions of microsynteny were identified in all of the sequenced tobacco species. Expression analysis showed that NtabGRF genes were highly expressed in actively growing tissues and responded to various hormone treatments. Our results provide foundational information about the GRF gene family in tobacco species, and open the door for future research on the functions of these genes.

Genome-wide analysis and expression profiling of the GRF gene family in oilseed rape (Brassica napus L.)

Growth regulating-factors (GRFs) are plant-specific transcription factors that help regulate plant growth and development. Genome-wide identification and evolutionary analyses of GRF gene families have been performed in Arabidopsis thaliana, Zea mays, Oryza sativa, and Brassica rapa, but a comprehensive analysis of the GRF gene family in oilseed rape (Brassica napus) has not yet been reported. In the current study, we identified 35 members of the BnGRF family in B. napus. We analyzed the chromosomal distribution, phylogenetic relationships (Bayesian Inference and Neighbor Joining method), gene structures, and motifs of the BnGRF family members, as well as the cis-acting regulatory elements in their promoters. We also analyzed the expression patterns of 15 randomly selected BnGRF genes in various tissues and in plant varieties with different harvest indices and gibberellic acid (GA) responses. The expression levels of BnGRFs under GA treatment suggested the presence of possible negative feedback regulation. The evolutionary patterns and expression profiles of BnGRFs uncovered in this study increase our understanding of the important roles played by these genes in oilseed rape.

Comparative Genomic Analysis of the GRF Genes in Chinese Pear (Pyrus bretschneideri Rehd), Poplar (Populous), Grape (Vitis vinifera), Arabidopsis and Rice (Oryza sativa).

Growth-regulating factors (GRFs) are plant-specific transcription factors that have important functions in regulating plant growth and development. Previous studies on GRF family members focused either on a single or a small set of genes. Here, a comparative genomic analysis of the GRF gene family was performed in poplar (a model tree species), Arabidopsis (a model plant for annual herbaceous dicots), grape (one model plant for perennial dicots), rice (a model plant for monocots) and Chinese pear (one of the economical fruit crops). In total, 58 GRF genes were identified, 12 genes in rice (Oryza sativa), 8 genes in grape (Vitis vinifera), 9 genes in Arabidopsis thaliana, 19 genes in poplar (Populus trichocarpa) and 10 genes in Chinese pear (Pyrus bretschneideri). The GRF genes were divided into five subfamilies based on the phylogenetic analysis, which was supported by their structural analysis. Furthermore, microsynteny analysis indicated that highly conserved regions of microsynteny were identified in all of the five species tested. And Ka/Ks analysis revealed that purifying selection plays an important role in the maintenance of GRF genes. Our results provide basic information on GRF genes in five plant species and lay the foundation for future research on the functions of these genes.

Genome-wide identification and analysis of the growth-regulating factor family in Chinese cabbage (Brassica rapa L. ssp. pekinensis).

BackgroundGrowth regulating factors (GRFs) have been shown to play important roles in plant growth and development. GRF genes represent a large multigene family in plants. Recently, genome-wide structural and evolutionary analyses of the GRF gene families in Arabidopsis, rice, and maize have been reported. Chinese cabbage (Brassica rapa L. ssp. pekinensis) is one of the most important vegetables for agricultural production, and a full genome assembly for this plant has recently been released. However, to our knowledge, the GRF gene family from Chinese cabbage has not been characterized in detail.ResultsIn this study, genome-wide analysis was carried out to identify all the GRF genes in Chinese cabbage. Based on the complete Chinese cabbage genome sequence, 17 nonredundant GRF genes, named BrGRFs, were identified and classified into six groups. Phylogenetic analysis of the translated GRF protein sequences from Chinese cabbage, Arabidopsis, and rice indicated that the Chinese cabbage GRF proteins were more closely related to the GRF proteins of Arabidopsis than to those of rice. Expression profile analysis showed that the BrGRF genes had uneven transcript levels in different organs or tissues, and the transcription of most BrGRF genes was induced by gibberellic acid (GA3) treatment. Additionally, over-expression of BrGRF8 in transgenic Arabidopsis plants increased the sizes of the leaves and other organs by regulation of cell proliferation.ConclusionsThe data obtained from this investigation will contribute to a better understanding of the characteristics of the GRF gene family in Chinese cabbage, and provide a basis for further studies to investigate GRF protein function during development as well as for Chinese cabbage-breeding programs to improve yield and/or head size.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-807) contains supplementary material, which is available to authorized users.

Genome-wide identification and analysis of growth regulating factor genes in Brachypodium distachyon: in silico approaches

Growth-regulating factor (GRF) genes may play important roles for regulating growth and development in different plant tissues and organs. Here we report the first genome-wide analysis of the GRF gene family in Brachypodium. We performed in silico comparative analysis of GRF genes, including their structure, duplication in the genome, conserved motifs, and phylogenetic relationship. At the end of the study, 10 BdGRF genes were identified. The highest number of GRF genes was identified on chromosome 1 with 5 members, whereas the least number of genes (only 1 member) was found on chromosomes 2, 4, and 5. Of those, a single segmental duplication was observed in the Brachypodium genome. Average exon and intron numbers were determined as 3 and 4, respectively. Motif analysis showed that WRC and QLQ residues were consistent in all GRF protein sequences. Gene Ontology terms showed that 10 BdGRF proteins grouped in the same biological function, biological process, and cellular component groups. In addition, we compared the new BdGRF proteins with the other monocot and dicot GRF proteins sequences. Phylogenetic analysis revealed that GRF proteins of monocot and dicot species were clustered together in a joined tree; in particular, the monocot species (Brachypodium, maize, and rice) were grouped into the same cluster with high bootstrap values. We assume that the results of this study will provide molecular insights about GRF proteins in grass species.