Somatic Embryogenesis Receptor-like Kinase Gene Research Articles

SERK genes identification and expression analysis during somatic embryogenesis and sporogenesis of sexual and apomictic Brachiaria brizantha (Syn. Urochloa brizantha).

In Brachiaria brizantha BbrizSERK1, BbrizSERK2 and BbrizSERK3 were identified. SERK expression marks somatic embryogenesis, sexual MMC, and sexual and apomictic PMC. BbrizSERK3 might have a regulatory role in reproductive development. Somatic embryogenesis receptor-like kinase (SERK) consists of plasma membrane receptor genes that have been characterized in various species, associated with several aspects of plant development, including reproduction. SERK genes are involved in anther development and in early embryo development in sexual and asexual seed formation. To comprehend the complexity of the SERK genes and their function in Brachiaria reproduction, we performed a homology-based search in a genomic database of a sexual B. brizantha and identified sequences of three SERK genes, BbrizSERK1, BbrizSERK2, and BbrizSERK3. RNASeq data showed equivalent abundance of BbrizSERK1 and BbrizSERK2 transcripts in ovaries at early megasporogenesis of sexuals and apomicts, while BbrizSERK3 transcripts were more abundant in ovaries of sexuals than in apomicts. BbrizSERK3 results in three coding sequences due to alternative splicing, among them Variant 1 results in a protein with all the predicted domains of a SERK. BbrizSERK transcripts were detected in male reproductive tissues of both sexual and apomictic plants, suggesting a role in controlling anther development. BbrizSERK transcripts were detected early in ovule development, in the integuments, and in the megaspore mother cell of the sexual plant, butnot in the cells that give rise to apomictic embryo sacs, suggesting a role in female reproductive development of sexuals. This paper provides evidences that SERK genes plays a role in the onset and establishment of somatic embryogenesis and in the reproductive development of B. brizantha and suggests a distinct role of BbrizSERK in apomixis initiation.

Multi-tasking of SERK-like kinases in plant embryogenesis, growth, and development: current advances and biotechnological applications

In plants, exogenous signals play a vital role in cell metabolism modification leading to growth and defense responses. The SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) belongs to a family of leucine-rich repeat receptor-like kinases (LRR-RLKs), involved in cell-to-embryo-transition, plant environmental responses, and plant development. SERKs interact with various ligand-binding receptors with complex-signaling networks and appear to function in diverse biological processes in plant development and physiology. The present review explores the current status of the role of SERK genes as candidate marker during plant embryogenesis. Furthermore, we describe recent advances in newly identified SERK functions and provide novel insights into different biotechnological advances. The recent advancements in newly identified SERK will help to unlock the long-standing mysteries of different biological and molecular mechanisms of plant cells.

Expression and DNA methylation of SERK, BBM, LEC2 and WUS genes in in vitro cultures of Boesenbergia rotunda (L.) Mansf.

The process of somatic embryogenesis and plant regeneration involve changes in gene expression and have been associated with changes in DNA methylation. Here, we report the expression and DNA methylation patterns of SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK), BABY BOOM (BBM), LEAFY COTYLEDON 2 (LEC2) and WUSCHEL (WUS) in meristematic block of newly emerged shoots from rhizome, embryogenic and non-embryogenic calli, prolonged cell suspension culture, ex vitro leaf, and in vitro leaf of regenerated plants of Boesenbergia rotunda. Among all seven samples, based on qRT-PCR, the highest level of expression of SERK, BBM and LEC2 was in embryogenic callus, while WUS was most highly expressed in meristematic block tissue followed by embryogenic callus. Relatively lower expression was observed in cell suspension culture and watery callus for SERK, LEC2 and WUS and in in vitro leaf for BBM. For gene specific methylation determined by bisulfite sequencing data, embryogenic callus samples had the lowest levels of DNA methylation at CG, CHG and CHH contexts of SERK, LEC2 and WUS. We observed negative correlation between DNA methylation at the CG and CHG contexts and the expression levels of SERK, BBM, LEC2 and WUS. Based on our results, we suggest that relatively higher expression and lower level of DNA methylation of SERK, BBM, LEC2 and WUS are associated with somatic embryogenesis and plant regeneration in B. rotunda.

Cellular and molecular changes associated with competence acquisition during passion fruit somatic embryogenesis: ultrastructural characterization and analysis of SERK gene expression.

The integration of cellular and molecular data is essential for understanding the mechanisms involved in the acquisition of competence by plant somatic cells and the cytological changes that underlie this process. In the present study, we investigated the dynamics and fate of Passiflora edulis Sims cotyledon explants that were committed to somatic embryogenesis by characterizing the associated ultrastructural events and analysing the expression of a putative P. edulis ortholog of the Somatic Embryogenesis Receptor-like Kinase (SERK) gene. Embryogenic calli were obtained from zygotic embryo explants cultured on Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzyladenine. Callus formation was initiated by the division of cells derived from the protodermal and subprotodermal cells on the abaxial side of the cotyledons. The isodiametric protodermal cells of the cotyledon explants adopted a columnar shape and became meristematic at the onset of PeSERK expression, which was not initially detected in explant cells. Therefore, we propose that these changes represent the first observable steps towards the acquisition of a competent state within this regeneration system. PeSERK expression was limited to the early stages of somatic embryogenesis; the expression of this gene was confined to proembryogenic zones and was absent in the embryos after the globular stage. Our data also demonstrated that the dynamics of the mobilization of reserve compounds correlated with the differentiation of the embryogenic callus.

High expression of SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE coincides with initiation of various developmental pathways in in vitro culture of Trifolium nigrescens.

The aim of this study was to identify and examine the expression pattern of the ortholog of SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE gene from Trifolium nigrescens (TnSERK) in embryogenic and non-regenerative cultures of immature cotyledonary-stage zygotic embryos (CsZEs). In the presence of 1-naphthaleneacetic acid and N6-[2-isopentenyl]-adenine, the CsZE regenerated embryoids directly and in a lengthy culture produced callus which was embryogenic or remained non-regenerative. As revealed by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), the TnSERK was expressed in both embryogenic and non-regenerative cultures, but the expression level was significantly higher in embryogenic ones. An in situ RNA hybridization assay revealed that the expression of TnSERK preceded the induction of cell division in explants, and then, it was maintained exclusively in actively dividing cells from which embryoids, embryo-like structures (ELSs), callus or tracheary elements were produced. However, the cells involved in different morphogenic events differed in intensity of hybridization signal which was the highest in embryogenic cells. The TnSERK was up-regulated during the development of embryoids, but in cotyledonary embryos, it was preferentially expressed in the regions of the apical meristems. The occurrence of morphological and anatomical abnormalities in embryoid development was preceded by a decline in TnSERK expression, and this coincided with the parenchymatization of the ground tissue in developing ELSs. TnSERK was also down-regulated during the maturation of parenchyma and xylem elements in CsZE and callus. Altogether, these data suggest the involvement of TnSERK in the induction of various developmental programs related to differentiation/transdifferentiation and totipotent state of cell(s).Electronic supplementary materialThe online version of this article (doi:10.1007/s00709-015-0814-5) contains supplementary material, which is available to authorized users.

Isolation and Expression Analysis of a SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) Gene in Curcuma alismatifolia Gagnep.

Somatic embryogenesis provides a useful tool to facilitate efficient mass propagation in plants. The SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) gene serves a fundamentally important role in somatic embryogenesis of many plant species. The isolation of a SERK gene homolog, namely CaSERK, from Curcuma alismatifolia Gagnep. cv. Blue Tung, was reported. Prediction of coding sequence showed that it encoded a protein of 628 amino acids showing high similarity to previously characterized SERK sequences and containing all the features shared by members of the SERK family, including five leucine-rich repeats and the distinctive proline-rich SPP domain. Investigation of CaSERK expression revealed that its transcripts were found throughout the whole somatic embryogenesis process with highest abundance in embryogenic callus. These results indicate that CaSERK might have somatic embryogenesis-associated functions in this economically important ornamental ginger. Detection of CaSERK transcript accumulation in flower and coma bract tissues is suggestive of its additional roles in other developmental signaling pathways.

Characterization of the third SERK gene in pineapple (Ananas comosus) and analysis of its expression and autophosphorylation activity in vitro.

Two somatic embryogenesis receptor-like kinase genes (identified as AcSERK1 and AcSERK2) have previously been characterized from pineapple (Ananas comosus). In this work, we describe the characterization of a third gene (AcSERK3) in this family. AcSERK3 had all the characteristic domains and shared extensive sequence homology with other plant SERKs. AcSERK3 expression was studied by in situ hybridization and quantitative real-time PCR to analyze its function. Intense in situ hybridization signals were observed only in single competent cells and competent cell clusters; no hybridization signal was detected in the subsequent stages of somatic embryogenesis. AcSERK3 was highly expressed in embryogenic callus compared to other organs, e.g., 20-80 fold more than in anther but similar to that of non-embryogenic callus, which was 20-50 fold that of anther. AcSERK3 expression in root was 80 fold higher than in anther and the highest amongst all organs tested. These results indicate that AcSERK3 plays an important role in callus proliferation and root development. His-tagged AcSERK3 protein was successfully expressed and the luminescence of His6-AcSERK3 protein was only ∼5% of that of inactivated AcSERK3 protein and reaction buffer without protein, and 11.3% of that of an extract of host Escherichia coli pET-30a. This finding confirmed that the AcSERK3 fusion protein had autophosphorylation activity.

Characterization and expression analysis of somatic embryogenesis receptor-like kinase genes from Phalaenopsis.

Somatic embryogenesis receptor-like kinase (SERK) genes have been found to be involved in the somatic embryogenesis of several plant species. We identified and characterized 5 PhSERK genes in the Phalaenopsis orchid. The amino acid sequences of PhSERKs and other SERK proteins are highly conserved, with the highest homology observed in the leucine-rich repeat-receptor-like kinase domain. All 5 PhSERKs were expressed in all Phalaenopsis organs examined (root, leaf, shoot apical meristem, and flower), with the strongest expression, particularly for PhSERK1 and 3, in the shoot apical meristem of mature plants. Expression of all PhSERKs was downregulated during early floral bud development and was upregulated gradually until the semi-open flower stage was reached. All 5 PhSERKs were expressed during both seed germination and protocorm-like-body (PLB) development. In germinated seeds, quantitative real-time PCR revealed upregulation of all PhSERKs except PhSERK4 at 1 week and downregulation after 4 weeks. The 5 PhSERKs were differentially expressed in the early stage of PLB development and maintained substantial levels during PLB formation, with PhSERK1 and 5 upregulated 1 week after culture and PhSERK2, 3, and 4 downregulated over this period. Because physical wounding of PLB stimulates secondary PLB formation, the PhSERK5 expression peak at week 3 coincided with visible and fully developed secondary PLBs. PhSERK5 may be important in PLB induction and subsequent development. Our PhSERK expression analysis revealed that these genes have a broad role during orchid plant development.

Characterization and expression analysis of AcSERK2, a somatic embryogenesis and stress resistance related gene in pineapple

The isolation and characterization of AcSERK2, another somatic embryogenesis receptor-like kinase (SERK) gene homolog, confirmed that there was a SERK gene family in pineapple. AcSERK2 shared high similarity to AcSERK1 and other SERKs, containing all the characteristic domains of the SERK family. The expressed recombinant protein of pET-AcSERK2 was soluble. AcSERK2 expression was effectively induced by 2,4-dichlorophenoxyacetic acid (2,4-D) and maintained in high level at the early stage of somatic embryogenesis (SE). The results of in situ hybridization showed that AcSERK2 expressed at high level in the competent cells under the induction of 2, 4-D and at low level during the development of meristematic center to globular embryo. This indicated that AcSERK2 could be used as a potential marker gene to monitor the acquisition of embryogenic competence. However, AcSERK2 expression was also detected in all the tested organs and the expression level decreased along with the development of calyx, ovule and ovary. These results suggested that AcSERK2 maybe play a broader role in organ development. AcSERK2 transcription could be induced by exogenous hormones and abiotic stress. It was deduced that AcSERK2 may participate in response to exogenous hormones and abiotic stress regulation in pineapple. The expression pattern of AcSERK2 during SE differed from that of AcSERK1, and this indicated that the SERK genes may be non-redundant in somatic embryogenesis.

Molecular characterization of a Cyrtochilum loxense Somatic Embryogenesis Receptor-like Kinase (SERK) gene expressed during somatic embryogenesis

Somatic embryogenesis is crucial for the propagation of endangered Ecuadorian orchid species, among them Cyrtochilum loxense, in view of the fact that their number in nature or in collections is quite reduced. One of the genes expressed during somatic and zygotic embryogenesis is Somatic Embryogenesis Receptor-like Kinase (SERK). Despite the development of somatic embryogenesis protocols for orchids, no SERK genes have been isolated from this family. This is the first report on the isolation of a full-length orchid SERK sequence, namely that of Cyrtochilum loxense (ClSERK). The identity of ClSERK was inferred by the presence of all domains typical of SERK proteins: a signal peptide, a leucine zipper domain, five LRRs, a serine proline-rich domain, a transmembrane domain, a kinase domain, and the C-terminal region. We have observed that the ClSERK gene is highly expressed in embryogenic calluses generated from protocorms at the time of appearance of embryonic morphological features. At later stages when embryos become well visible on calluses, ClSERK gene expression decreases. Compared to early stages of embryo formation on calluses, the expression detected in leaf tissue is far lower, thus suggesting a role of this gene during development.

Two SERK genes are markers of pluripotency in Cyclamen persicum Mill.

The genetic basis of stem cell specification in somatic embryogenesis and organogenesis is still obscure. SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) genes are involved in embryogenesis and organogenesis in numerous species. In vitro culture of Cyclamen persicum immature ovules provides a system for investigating stem cell formation and maintenance, because lines forming either organs or embryos or callus without organs/embryos are available for the same cultivar and plant growth regulator conditions. The present aim was to exploit this property of cyclamen cultures to understand the role of SERK(s) in stem cell formation and maintenance in somatic embryogenesis and organogenesis in vitro, in comparison with expression in planta. CpSERK1 and CpSERK2 were isolated from embryogenic callus. CpSERK1 and CpSERK2 levels by RT-PCR showed that expression is high in embryogenic, moderate in organogenic, and null in recalcitrant calli. in situ hybridizations showed that the expression of both genes started in clumps of pluripotent stem cells, from which both pre-embryogenic aggregates and organ meristemoids derived, and continued in their trans-amplifying, meristem-like, derivatives. Expression declined in organ meristemoids, in parallel with a partial loss of meristematization. In mature somatic embryos, and in shoot and root primordia, CpSERK1 and CpSERK2 were expressed in meristems, and similar patterns occurred in zygotic embryo and primary meristems in planta. The results point to SERK1 and SERK2 as markers of pluripotency in cyclamen. It is proposed that the high expression of these genes in the trans-amplifying derivatives of the stem cells maintains a pluripotent condition leading to totipotency and, consequently, somatic embryogenesis.

Cloning and Molecular Characterization of a SERK Gene Transcriptionally Induced During Somatic Embryogenesis in Ananas comosus cv. Shenwan

A somatic embryogenesis receptor-like kinase (SERK) gene, designated as AcSERK1, was isolated from pineapple (Ananas comosus cv. Shenwan). AcSERK1 shared all the characteristic domains of the SERK family, including five leucine-rich repeats, one proline-rich region motif, transmembrane domain, and kinase domains. Somatic embryogenic cultures of pineapple were established following transfer of callus cultures to Murashige and Skoog (1962) medium containing 2,4-dichlorophenoxyacetic acid. The role of AcSERK1 during establishment of somatic embryogenesis in culture was investigated. The AcSERK1 was highly expressed during embryogenic competence acquisition and global embryo formation in culture. These findings were obtained along with morphological changes in callus cultures exhibiting embryogenic potential. Overall, levels of expression of AcSERK1 were lower in nonembryogenic tissues and organs than in embryogenic callus. In situ hybridization analysis revealed that AcSERK1 expression was detected in embryogenic tissues, including single competent cells, meristematic centers wherein embryogenic structures are formed, and global embryos. These results suggested that AcSERK1 expression was associated with induction of somatic embryogenesis and that it could be used as a potential marker gene to monitor the transition of pineapple callus tissues into competent and embryogenic cells and tissues.

Isolation and Functional Characterization of a SERK Gene from Soybean (Glycine max (L.) Merr.)

It is well accepted that somatic embryogenesis serves a primary role in plant regeneration. However, it is also a model system to explore the regulatory and morphogenetic events in the life of a plant. To date, a suite of genes that serve important roles in somatic embryogenesis have been isolated and identified. In the present study, a novel gene designated as GmSERK1 was isolated from soybean (Glycine max (L.) Merr). Sequence and structural analysis determined that the GmSERK1 protein, which encodes 624 amino acids, belongs to the somatic embryogenesis receptor-like kinase (SERK) gene family. GmSERK1 shared all the characteristic domains of the SERK family, including five leucine-rich repeats, one proline-rich region motif, transmembrane domain, and kinase domains. DNA gel blot analysis indicated that a single copy of the GmSERK1 gene resides in the soybean genome. The GmSERK1 tissue-specific and induced expression patterns were explored using quantitative real-time PCR. Dissimilar expression levels in various tissues under different treatments were found. In addition, transient expression experiments in onion epidermal cells indicated that the GmSERK1 protein was located on the plasma membrane. The results from this study suggested that GmSERK1, a member of the SERK gene family, exhibits a broader role in various aspects of plant development and function, in addition to its basic functions in somatic embryogenesis.

Cloning, molecular characterization and expression analysis of a SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE gene (CitSERK1-like) in Valencia sweet orange

Somatic embryogenesis receptor-like kinase (SERK) belonging to the receptor-like kinases (RLKs) has been shown to be implicated in somatic embryogenesis (SE). In this study, a somatic embryogenesis receptor-like gene CitSERK1-like was cloned and characterized from Citrus sinensis cv. ‘Valencia’, a genotype with high somatic embryogenesis capacity for over 26 years. Fifteen consecutive amino acids in putative leucine zipper domain of CitSERK1-like gene were different from the reported CitSERK1 gene. Homology search and sequence analysis demonstrated that the deduced CitSERK1-like protein shared a high degree of identity with SERKs from other species in sequence and structure. Real-time PCR analysis revealed that the transcript of CitSERK1-like was enhanced during the induction of SE. At subsequent embryo-transition phase, a moderate level of expression was detected in heart-torpedo and cotyledon embryos, while low expression was detected in globular embryo. Among the different tissues, the expression of CitSERK1-like was highest in young leaves. Further analysis of its spatial expression by in situ hybridization revealed that CitSERK1-like was mainly located in the embryogenic callus and vascular cells of different embryos or tissues. The results of temporal and spatial expression of CitSERK1-like showed that it played critical roles throughout the process of SE and had a broader role in plant development. In addition, CitSERK1-like expression was up-regulated by 2, 4-D and NAA at the early stage, but down-regulated afterward. Taken together, it suggested that CitSERK1-like activated complex developmental pathways associated with somatic embryogenesis and plant growth.

Structural Characterization and Expression Analysis of the SERK/SERL Gene Family in Rice (Oryza sativa)

Somatic embryogenesis (SE) is the developmental restructuring of somatic cells towards the embryogenic pathway and forms the basis of cellular totipotency in angiosperms. With the availability of full-length cDNA sequences from Knowledge-based Oryza Molecular Biological Encylopedia (KOME), we identified the leucine-rich repeat receptor-like kinase (LRR-RLK) genes from rice (Oryza sativa), which also encompasses genes involved in regulating somatic embryogenesis. Eight out of eleven of the rice SERK and SERL (SERK-like) genes have the TIGR annotation as (putative) brassinosteroid insensitive 1-associated receptor kinase (precursor). Real-time polymerase chain reaction analysis was undertaken to quantify transcript levels of these 11 genes. Most of these genes were upregulated by brassinosteroids although only a few of these displayed auxin induction. The expression profile of these genes is nearly uniform in the zygotic embryogenic tissue, but the expression pattern is more complex in the somatic embryogenic tissue. It is likely that OsSERKs and OsSERLs may be involved in somatic embryogenesis and also perform a role in morphogenesis and various other plant developmental processes. Functional validation of these somatic embryogenesis receptor-like kinase genes may help in elucidating their precise functions in regulating various facets of plant development.

Suppression of SERK gene expression affects fungus tolerance and somatic embryogenesis in transgenic lettuce.

The Somatic embryogenesis receptor-like kinase (SERK) gene plays an important role in plant somatic and zygotic embryogenesis induction. The gene encodes an LRR-containing receptor-like kinase protein. Studies have been carried out focusing on different aspects of its function, but definitive conclusions on its role are far from being reached. SERK expression is generally detected in cells in which somatic or zygotic embryogenesis has been triggered. Transgenic lettuce lines were produced to silence the endogenous SERK gene using antisense RNA. The average number of seeds per flower in the R(1) and R(2) generations was similar for both transgenic and non-transgenic lines. However, a reduction in the number of viable grained seeds was observed in four studied transgenic lines. Endogenous SERK expression analysis revealed the absence of detectable LsSERK gene transcripts in three transgenic lines, which presented a reduction in their ability to form in vitro somatic embryonic structures. In addition, transgenic lines showed enhanced susceptibility to the pathogenic fungus Sclerotinia sclerotiorum, when compared to control plants. The results support the idea that SERK genes might not only be involved in plant growth and development, but probably also in a general mechanism of biotic and abiotic stress perception.

Cloning and molecular characterisation of a potato SERK gene transcriptionally induced during initiation of somatic embryogenesis

Somatic embryogenesis offers great potential in plant propagation, long-term germplasm conservation, and as a suitable model system for deciphering early events during embryogenesis. The up-regulation and ectopic expression of a SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) gene has been shown to mark and enhance embryogenic competence in somatic cells of model plant species. We have cloned and characterised a SERK gene (StSERK1) from potato (Solanum tuberosum L.), an important crop plant. Sequence analysis of StSERK1 revealed high levels of similarity to other plant SERKs, as well as a conserved intron/exon structure which is unique to members of the SERK family. Furthermore, StSERK clustered most closely with SERK gene family members such as MtSERK1, CuSERK1, AtSERK1, and DcSERK, implicated in evoking somatic embryogenesis. Monitoring of SERK expression during progression of potato somatic embryogenesis revealed increased StSERK expression during the induction phase. Subsequently, during the embryo transition phases, StSERK expression was unchanged and did not vary among embryo-forming and inhibitory conditions. However, in isolated somatic embryos StSERK expression was again up-regulated. In other plant parts (leaves, true potato seeds, microtubers and flower buds), StSERK showed different levels of expression. Expression analysis suggests that the isolated StSERK could be a functional SERK orthologue. The possible role of SERK as a marker of pluripotency, rather than embryogenesis alone, is discussed.

Advanced study on SERK genes family

Somatic embryogenesis receptor-like kinase (SERK) genes were identified in different plant species, such as Daucus carota, Arabidopsis thaliana and Oryza sativa. Studies of these genes showed that they have conserved structures and expression patterns. These genes are not only expressed in embryogenic tissues, but also in post-embryogenic development. They function in embryogenesis and reproductive development and defences in particular, related signal transduction pathways.

Isolation and characterization of the somatic embryogenesis receptor-like kinase gene homologue (CitSERK1) from Citrus unshiu Marc.

A cDNA clone was isolated for a somatic embryogenesis receptor-like kinase (SERK) gene homologue (CitSERK1) from Citrus unshiu Marc. CitSERK1 revealed high amino acid sequence homologies to the past isolated SERK genes of Arabidopsis thaliana, Zea mays, and Daucus carota and had the common structural features of a putative leucine zipper (ZIP) with five repeat units in the extracellular domain, a proline-rich region (SPP) just upstream of the transmembrane domain, and the kinase domain in the C-terminal. The mRNA expression of CitSERK1 was relatively uniform in all plant tissues except those of mature fruit at days after flowering (DAF) 180 and abundant around the stage of nucleous division and enlargement. During callus culture, the transcripts of CitSERK1 were less than the detectable levels in proliferating non-embryogenic calli but increased until they could be detected in the calli after the initiation of embryogenesis, suggesting that CitSERK1 could be involved in embryogenesis or embryo development. In addition, genetic linkage analysis showed that the SERK1 locus was independent of both loci concerning two traits, polyembryony and seedlessness derived from Satsuma mandarin, which have been considered to be highly important characteristics in Citrus genetic improvement.

Spatial expression of a sunflower SERK gene during induction of somatic embryogenesis and shoot organogenesis

Organogenesis or somatic embryogenesis can be induced on immature zygotic embryos (IZE) of sunflower depending on the culture conditions. Both morphogenic processes originate from the same group of cells and show identical kinetics. Using real-time PCR and in situ hybridisation, we showed that somatic embryogenesis receptor-like kinase ( SERK) transcripts accumulate early after the beginning of the culture in the morphogenic zone of IZE explants whatever the induction conditions used, i.e. organogenic, embryogenic or highly embryogenic conditions. Quantitative analyses failed to show any correlation between the SERK expression level during the period decisive for the orientation of the morphogenic pathway, i.e. the first 2 days of culture, and the type of morphogenesis induced. However, after 2 days of culture on the organogenic medium, the SERK gene expression level was severely down-regulated in the IZE explants. At 4 days of culture, SERK transcripts were no longer detectable by in situ hybridisation in the developing shoot structures whereas they still continued to accumulate in the embryonic structures induced on both embryogenic and highly embryogenic culture media. The significance of these expression analyses was addressed by transfer medium experiments. Results revealed that IZE cultured on the organogenic medium were able to form somatic embryos when transferred on the highly embryogenic medium as long as the SERK transcripts accumulated at a high level in their morphogenic zone, i.e. first 2 days of culture. Passt this delay, explants rapidly lost their embryogenic competence. Indeed, after 4 days of culture on the organogenic medium, IZE were definitely oriented towards shoot organogenesis. Taken together, these data suggest that reactive cells of IZE develop the competence to somatic embryogenesis during the first day of culture whatever the morphogenic induction conditions used.