Embryo Formation Research Articles

Abnormal BMI in Male and/or Female Partners Are Deleterious for Embryonic Development and Pregnancy Outcome During ART Process: A Retrospective Study

ObjectiveTo investigate the effect of BMI in male and/or female partners on embryo development and clinical pregnancy outcome during ART.MethodsData of 11,130 cycles between January 2018 and December 2020 were retrospectively analyzed. They were divided into Group A, B, C, and D based on couples’ BMI values, also were divided into Group I, II, III and IV in IVF cycles and Group i, ii, iii, and iv in ICSI cycles. After grouping, inter-group indicators comparisons and logistic regression analysis were performed.ResultsIn IVF cycles, CPR in Group I and Group III were higher than Group IV. In Group III, it was higher than Group II. The AR in Group IV was higher, but the LBR was lower than Group I, Group II, and Group III. Logistic regression analysis results suggested that AR in Group IV was higher than that in Group I in IVF cycles, whereas LBR was lower.In ICSI cycles, high-quality embryo rate in Group i and Group ii were both higher than that in Group iii and Group iv. The CPR in Group i was higher than Group ii and Group iv, and in Group iii was higher than Group ii and Group iv. The AR in Group i was lower than Group iii and Group iv, and AR in Group ii was lower than Group iv. LBR, in Group I it was higher than Group ii, Group iii, and Group iv. Logistic regression analysis results suggested CPR in Group ii was significantly lower than that in Group i. AR in Group iii was considerably higher than that in Group i. LBR in Group ii and Group iv were significantly lower than that in Group i.ConclusionFemale higher BMI was not conducive to the formation of high-quality embryos in ICSI cycle. Female and/or male BMI affected AR and LBR more than CPR not only in IVF cycles, but also in ICSI cycles.

PEG induces maturation of somatic embryos of Passiflora edulis Sims ‘UENF Rio Dourado’ by differential accumulation of proteins and modulation of endogenous contents of free polyamines

Sour passion fruit (Passiflora edulis Sims) has economic and social relevance and is an alternative crop mainly for family farming agriculture. The aim of this work was to evaluate the influence of polyethylene glycol (PEG) on the maturation of somatic embryos associated with differential accumulation of proteins and changes in the endogenous polyamine (PA) content during somatic embryogenesis of P. edulis ‘UENF Rio Dourado’. Maturation of somatic embryos was performed using embryogenic callus in MS culture medium with PEG 6% or without PEG (control). PEG 6% promoted the maturation of a significantly higher number of somatic embryos at globular and cotyledonary stages when compared to the control treatment. The higher somatic embryo formation induced by PEG 6% was associated with an increase in endogenous contents of free spermine, a PA with an important role in the maturation process of somatic embryogenesis cultures. Comparative proteomic analyses of PEG 6%/control revealed that PEG 6% treatment induced the up-accumulation of proteins related to the ATP metabolic, glycolytic, generation of precursor metabolite energy, and the response to light stimulus processes. The down-accumulated proteins were related mainly to the cellular metabolic process. The use of PEG induced the maturation and development of somatic embryos of P. edulis Sims ‘UENF Rio Dourado’ by the differential accumulation of proteins and modulation of endogenous contents of PAs.

Wuschel2 enables highly efficient CRISPR/Cas-targeted genome editing during rapid de novo shoot regeneration in sorghum

For many important crops including sorghum, use of CRISPR/Cas technology is limited not only by the delivery of the gene-modification components into a plant cell, but also by the ability to regenerate a fertile plant from the engineered cell through tissue culture. Here, we report that Wuschel2 (Wus2)-enabled transformation increases not only the transformation efficiency, but also the CRISPR/Cas-targeted genome editing frequency in sorghum (Sorghum bicolor L.). Using Agrobacterium-mediated transformation, we have demonstrated Wus2-induced direct somatic embryo formation and regeneration, bypassing genotype-dependent callus formation and significantly shortening the tissue culture cycle time. This method also increased the regeneration capacity that resulted in higher transformation efficiency across different sorghum varieties. Subsequently, advanced excision systems and “altruistic” transformation technology have been developed to generate high-quality morphogenic gene-free and/or selectable marker-free sorghum events. Finally, we demonstrate up to 6.8-fold increase in CRISPR/Cas9-mediated gene dropout frequency using Wus2-enabled transformation, compared to without Wus2, across various targeted loci in different sorghum genotypes.

Effect of Sperm 26S Proteasome on Embryo Formation and Development in In Vitro Fertilization.

The aim of this study was to evaluate the effects of sperm proteasome activity on fertilization outcome and embryo development after IVF. Following density gradient centrifugation for IVF purpose, the spermatozoa of 84 infertile patients with tubal factor were evaluated by luciferase enzymatic activity to assess the proteasome quantity. The mean age of patients was 33.8years, and the mean concentration of human spermatozoa 26S proteasome was 674.53ng/ml. After IVF, the embryos were scored for morphology. The spermatozoa proteasome activity was both positively correlated with fertilization rate in vitro (P = 0.0003) and 2PN rate (P = 0.0007). Compared to low fertilization rate group, the high fertilization rate group showed a significantly higher level of spermatozoa proteasome activity (P = 0.002). In conclusion, sperm proteasome activity provides additional data on sperm functional capacity in terms of fertilization during IVF.

Application of the FISH Method for Analyzing Chromosome Segregation Patterns in Preimplantation Embryos from Robertsonian Translocation Carriers

Robertsonian translocations (RTs) are very common balanced structural chromosome rearrangements in humans. Due to alterations in the chromosome segregation pattern and the formation of unbalanced gametes and embryos, the carriers of RTs have a significant risk of reproductive failure. For over 30 years, fluorescent in situ hybridization (FISH) has been used for preimplantation genetic testing for chromosomal structural rearrangement (PGT-SR) in carriers of RTs. The data obtained by performing FISH for PGT-SR can be useful for analyzing segregation patterns in preimplantation embryos. We analyzed chromosome segregation patterns in 964 embryos from 100 couples who were carriers of various types of RTs by performing FISH or using the array comparative genomic hybridization (aCGH) method. When the carriers were male, the predominant segregation mode was alternate (detected in 42.4% of embryos), whereas, when the carriers were female, alternate and adjacent types of segregation were detected in most embryos (32.3% and 26.6% of embryos, respectively). About 33% of balanced/euploid embryos in IVF cycles were formed when the carriers of RT were male, and about 24% of such embryos were formed when the carriers were female. No association was found between sperm parameters and malsegregation rates in male carriers of RT. In this study, we found that female and male RT carriers have different reproductive risks associated with chromosome segregation patterns and the rate of balanced/euploid embryos.

Self-assembly growth of electrolytic silver dendrites

The atomic level assembly of silver dendrite has never been disclosed despite the numerous studies published on fractal dendrite structures. We report for the first time an HRTEM investigation of the formation of atomic embryos (< 5 nm) and the self-assembly of atoms on an atomic plane of the tip of a dendrite arm. The mechanism of dendrite formation proceeds via the sequence of amorphous embryos aggregates (5–10 nm), nuclei, crystallites (10–20 nm), dendritelets (50–100 nm) and submicron dendrite protypes. The atomic plane is an entirely atomic-level zig-zag structure with d-spacing kink steps. The zig-zag structure triggers the self-assembly of atoms and thus directional growth to produce a dendrite arm with a high aspect ratio.

Study of Various Floral Types on Different Plants of Garcinia indica (Thouars) Choisy and Correlation of Its Functionality in Sexual Reproduction

ABSTRACT Genus Garcinia shows high diversity of sexual systems; most of them are dioecious in nature. The other reported sexual systems of Garcinia include gynodioecious, androdioecious, monoecious, and andromonoecious. Garcinia indica (Thouars) Choisy was reported as facultative apomictic polygamodioecious species. There is a large diversity in floral types. Hence, a detailed study of various floral types found on different types of plants of G. indica and to correlate its functionality with sexual reproduction was undertaken. The study at Kokum Plot, BSSKVV, Dapoli, revealed that the male plants could be categorized into four types – androecious, functional males, andromonoecious, and hermaphrodites. The female plants displayed large variation in number and viability of stamens/staminodes. Each category showed significant difference in phenology, floral morphology and pollen types. In December, the percentage of staminate flowers (79.13 ± 2.93%) in androecious trees was significantly higher than functional males (58.61 ± 8.60%) or andromonoecious trees (56.36 ± 2.77%) (p ≤ .01). As female plants started flowering, all male morphotypes bore significantly higher numbers of staminate flowers and carpellate flowers were significantly reduced. Pollen viability was highest in androecious flowers at 90.11 ± 1.10% and lowest at 64.70 ± 2.09% in hermaphrodite trees. Earlier studies reported female flowers bore well-developed female gametophyte, but the embryo development was not observed. Thus, the fruit formation might be apomictic, but in present embryological investigation; more than 60% of female and andromonoecious flowers, a well-developed embryo was formed between 8 and 24 h after anthesis. Thus, the embryo formation might be sexual or apomictic. Floral types favoring sexual reproduction are essential for conservation of highly endemic species.

Liquid cell transmission electron microscopy reveals C-S-H growth mechanism during Portland cement hydration

We report the first application of in-situ liquid cell transmission electron microscopy (LC-TEM) to research hydration reactions of nano OPC, providing nanoscale insight into early reaction mechanisms. We demonstrate that the formation and growth of C-S-H precipitates starts through lateral growth of planar silicate sheets, but soon continues in all directions resulting in a 3D microstructure. Furthermore, nanocrystalline C-S-H structures with sizes between 5 nm to 10 nm were observed inside the amorphous or highly disordered C-S-H matrix, denoting that C-S-H growth is conformed to layered structure model. Crack formation and propagation inside C-S-H precipitates confirms the presence of increasing lattice strain due to growing defects that limits the growth of a fully crystalline structure by buckling and separating the sheets. The rolling up and crumbling of C-S-H sheets promotes the formation of new embryos, leading to the growth of precipitates in all direction and finally their coalescence.

EFFECT OF CARBENECILLIN AND CEFOTAXIME ON CALLUS INDUCTION AND DEVELOPMENT SOMATIC EMBRYOS OF MALAYSIAN INDICA RICE MR219

Carbenicillin and cefotaxime, two antibiotics routinely employed to exclude Agrobacterium tumefaciens during plant transformation, were studied for their effects on callus growth and the formation of somatic embryos. Antibiotics were tested to see how they responded to the regeneration of Malaysian indica rice cv. MR219. In a preliminary study, callus fresh weight gain appeared to be improved in media containing 200 mg/L carbenicillin and 300-400 mg/L cefotaxime but was severely hindered in media containing higher concentrations. Somatic embryos were discovered in greater numbers in the medium containing 200 mgl-1 carbenicillin or 300 mgl-1 cefotaxime, at 76 and 71, respectively. It was possible that these antibiotics showed a positive response on somatic embryos development when compared to those without antibiotics. Carbenicillin and cefotaxime concentrations greater than 300 mg/L should not be utilized for the process

Effect of ACY-1215 on cytoskeletal remodeling and histone acetylation in bovine somatic cell nuclear transfer embryos

The latest studies indicated that in addition to alterations in abnormal chromosome epigenetic modifications, the abnormal cytoskeletal changes are also an important cause for the developmental failure of somatic cell nuclear transfer (SCNT) embryos. In the present study, the effects of ACY-1215, a specific inhibitor of HDAC6, on the acetylation of α-tubulin, histone epigenetic modification, spindle formation and embryonic development of early bovine SCNT embryos were studied. The results showed that acetylation of α-tubulin, H3K9, and H4K16 was significantly lower in SCNT embryos than in vitro fertilization (IVF) embryos. After ACY-1215 treatment, the acetylation level of α-tubulin, H3K9, and H4K16 of SCNT embryos was closer to that of IVF embryos. ACY-1215 treatment reduced spindle abnormalities, delayed the time of first cleavage of embryos, increased the total cell number and trophectoderm cells numbers, and reduced apoptosis in SCNT blastocysts. ACY-1215 regulated the process of embryonic epigenetic modification and cytoskeletal protein acetylation, corrected abnormal development of SCNT embryos, and improved SCNT embryonic development potential.

Effects of genotype and nutrient medium on obtaining haploid plants through ovary culture in cucumber.

Cucumber is a species that breeding studies for variety development are carried out intensively. However, double haploidy technology, which aims to shorten the breeding process, has not yet reached the desired level. Three induction (M1: Murashige and Skoog (MS), 0.04mg L-1 Thidiazuron (TDZ); M2: MS, 0.15mg L-1 2,4-Dichlorophenoxyacetic acid (2,4-D), 1.5mg L-1 Kinetin; M3: MS, 0.1mg L-1 2,4-D, 1mg L-1 6-Benzylaminopurine (BAP) and one regeneration (MS, 0.2mg L-1 BAP, 0.05mg L-1 1-Naphthaleneacetic acid (NAA) media and 31 cucumber genotypes were used. At the end of study, in terms of embryo formation, M3 (33.41 embryos per 100 cultured ovaria, 99.61 embryos per 100 developed ovaria) and M2 (30.70 embryos per 100 cultured ovaria, 122.05 embryos per 100 developed ovaria) were found to be better than M1 (17.54 embryos per 100 cultured ovaria, 68.34 embryos per 100 developed ovaria). For plant formation, M1 (13.23 plants per 100 embryos) and M2 were found to be more succesful than M3. Ploidy analyses performed on 72 of 98 plants through flow cytrometry showed that obtained plants were various ploidy level (34.72% haploid, 37.5% diploid, 22.22% mixoploid, and 5.55% tetraploid). According to the results of the research, 2,4-D added to the nutrient media seems to be successful in induction of ovary culture in cucumber. In plants determined as diploid according to ploidy analysis, doubled haploid situation should be checked by molecular analysis.

Somatic embryogenesis: from discovery through investigation to application

Plant cells possess the remarkable ability to adapt to environmental changes. It is manifested by formation of embryos directly from the cells of plant body, bypassing the fertilization stage. These embryo structures develop into complete plants. The process itself, to distinguish the path of formation and emphasize consistency with zygotic embryogenesis, is referred to as somatic embryogenesis (SE). Although more than 60 years have passed since the first publication on the phenomenon has been written, the mechanism of reprogramming of a somatic cell into an embryogenic one is still not fully understood. This is a critical step in SE that can be induced by exo- and endogenous factors and stress treatments. The exposition of plant material to these factors affects the reorganization of the chromatin structure and gene expression, which can consequently trigger the program of embryogenesis. The paper reviews current knowledge on how the identity of totipotent cells is determined and the which stimuli are required to reprogram somatic cell development. Knowledge of key molecular regulators and the network of relationships that control the SE induction is summarized. Issues that are important for enhancing the understanding of the mechanisms underlying totipotency are also defined. Finally, the practical potential of SE is demonstrated, and examples of its use are provided.

Gene mining of 100‐grain weight and the number of four‐seed pods in soybean (Glycine max)

AbstractThe number of four‐seed pods and 100‐grain weight are important yield components in soybean. Typically, they are negatively correlated traits. Generally, soybean varieties with a high 100‐grain weight and a higher number of three‐ or four‐seed pods are more likely to obtain a high yield. It is difficult to select for double‐excellent traits that meet the needs of farmers and the market using conventional breeding methods. The purpose of this study was to mine genes associated with the number of four‐seed pods and 100‐grain weight in soybean. Whole‐transcriptome sequencing was performed on four specific chromosome segment substitution lines (HWMN, HWFN, LWMN and LWFN) combined with the distribution of blocks imported from wild soybean DNA fragments and gene annotation. The material was sampled in each of the eight development stages. Among them, globular embryo formation, heart embryo formation, and cotyledon primordium formation were used to mine differentially expressed genes (DEGs). A total of 3792 DEGs were identified, and 25 expression patterns were obtained by the K‐means rapid clustering method. GO enrichment analysis of DEGs was performed by Agrigo, and a total of 43 GO terms were enriched. Through annotation analysis, 126 DEGs associated with seed size and number were obtained. Combined with analysis of the introduced DNA fragments of wild soybean ZYD00006, 19 genes that eventually aligned on those blocks were obtained. Six representative genes were selected as candidate genes, namely, Glyma.07G218200, Glyma.05G162100, Glyma.19G240800, Glyma.04G180300, Glyma.05G214400 and Glyma.12G146700. They may regulate the formation of 100‐grain weight and the number of four‐seed pods, which will provide reference information for the genetic improvement of agronomic traits, such as seed number per pod and 100‐grain weight of soybean, in the future and will enrich the theoretical basis of high yield in soybean.

Biological Role of Nodal Modulator: A Comprehensive Review of the Last Two Decades.

Nodal modulator (NOMO) is a type I transmembrane protein that is conserved in various human tissues. Humans have three highly similar NOMO proteins, namely NOMO1, NOMO2, and NOMO3. These three proteins are closely related and may have similar functions. NOMO has been identified as a part of a protein complex that mediates a wide range of biological processes such as tumor formation, bone and cartilage formation, embryo formation, facial asymmetry, and development of congenital heart disease. To date, a few studies have focused on the role of NOMO; however, the mechanism underlying its effects remains unknown. To improve our understanding regarding NOMO, we reviewed the role of NOMO in different diseases and investigated the mechanism underlying its effects.

Direct visualization of pre-protamine 2 detects protamine assembly failures and predicts ICSI success.

Histone-to-protamine transition is an essential step in the generation of fully functional spermatozoa in various mammalian species. In human and mouse, one of the two protamine-encoding genes produces a precursor pre-protamine 2 (pre-PRM2) protein, which is then processed and assembled. Here, we design an original approach based on the generation of pre-PRM2-specific antibodies to visualize the unprocessed pre-PRM2 by microscopy, flow cytometry and immunoblotting. Using mouse models with characterized failures in histone-to-protamine replacement, we show that pre-PRM2 retention is tightly linked to impaired nucleosome disassembly. Additionally, in elongating/condensing spermatids, we observe that pre-PRM2 and transition protein are co-expressed spatiotemporally, and their physical interaction suggests that these proteins act simultaneously rather than successively during histone replacement. By using our anti-human pre-PRM2 antibody, we also measured pre-PRM2 retention rates in the spermatozoa from 49 men of a series of infertile couples undergoing ICSI, which shed new light on the debated relation between pre-PRM2 retention and sperm parameters. Finally, by monitoring 2-pronuclei embryo formation following ICSI, we evaluated the fertilization ability of the sperm in these 49 patients. Our results suggest that the extent of pre-PRM2 retention in sperm, rather than pre-PRM2 accumulation per se, is associated with fertilization failure. Hence, anti-pre-PRM2 antibodies are valuable tools that could be used in routine monitoring of sperm parameters in fertility clinics, as well as in experimental research programmes to better understand the obscure process of histone-to-protamine transition.

Overexpression of Oil Palm Early Nodulin 93 Protein Gene (EgENOD93) Enhances In Vitro Shoot Regeneration in Arabidopsis thaliana.

EgENOD93 was first identified in a cDNA microarray study of oil palm tissue culture where it was highly expressed in leaf explants with embryogenic potential. Functional characterization via an RNA interference study of its orthologue in Medicago truncatula demonstrated a significant role of this gene in somatic embryo formation. In this study, EgENOD93 was overexpressed in the important model plant Arabidopsis thaliana to investigate the embryogenic potential of EgENOD93 transgenic Arabidopsis explants compared to explants from control plants (pMDC140 and WT). Experiments using leaf explants revealed higher numbers of regenerated shoots at day 27 in all the homozygous transgenic Arabidopsis cultures (Tg01, Tg02 and Tg03) compared to controls. The expression level of EgENOD93 in Arabidopsis cultures was quantified using reverse transcription quantitative real-time PCR (RT-qPCR). The results supported the overexpression of this gene in transgenic Arabidopsis cultures, with 6 and 10 times higher expression of EgENOD93 in callus at Day 9 and Day 20, respectively. Overall, the results support the role of EgENOD93 in the enhancement of shoot regeneration in transgenic Arabidopsis. This together with the previous results observed in oil palm and Medicago truncatula suggests that ENOD93 plays a key role in the induction of somatic embryogenesis. A similarity to early nodulation-like ontogeny is possible.

Global Analysis of the WOX Transcription Factor Gene Family in Populus × xiaohei T. S. Hwang et Liang Reveals Their Stress−Responsive Patterns

The WUSCHEL−related homeobox (WOX) family is a group of plant−specific transcription factors that play important regulatory roles in embryo formation, stem cell stability, and organogenesis. To date, there are few studies on the molecular mechanisms involved in this family of genes in response to stress. Thus, in this study, eight WOX genes were obtained from an endemic Chinese resilient tree species, Populus × xiaohei T. S. Hwang et Liang. Bioinformatic analysis showed that the WOX genes all contained a conserved structural domain consisting of 60 amino acids, with some differences in physicochemical properties. Phylogenetic analysis revealed that WOX members were divided into three evolutionary clades, with four, one, and three members in the ancient, intermediate, and modern evolutionary clades, respectively. The conserved structural domain species as well as the organization and gene structure of WOX genes within the same subfamily were highly uniform. Chromosomal distribution and genome synteny analyses revealed seven segmental−duplicated gene pairs among the PsnWOX gene family that were mainly under purifying selection conditions. Semi−quantitative interpretation (SQ−PCR) analysis showed that the WOX gene was differentially expressed in different tissues, and it was hypothesized that the functions performed by different members were diverse. The family members were strongly and differentially expressed under CdCl2, NaCl, NaHCO3, and PEG treatments, suggesting that WOX genes function in various aspects of abiotic stress defense responses. These results provide a theoretical basis for investigating the morphogenetic effects and abiotic stress responses of this gene family in woody plants.

A single-cell RNA-seq analysis of Brachyury-expressing cell clusters suggests a morphogenesis-associated signal center of oral ectoderm in sea urchin embryos

Brachyury is a T-box family transcription factor and plays pivotal roles in morphogenesis. In sea urchin embryos, Brachyury is expressed in the invaginating endoderm, and in the oral ectoderm of the invaginating mouth opening. The oral ectoderm is hypothesized to serve as a signaling center for oral (ventral)-aboral (dorsal) axis formation and to function as a ventral organizer. Our previous results of a single-cell RNA-seq (scRNA-seq) atlas of early Strongylocentrotus purpuratus embryos categorized the constituent cells into 22 clusters, in which the endoderm consists of three clusters and the oral ectoderm four clusters (Foster et al., 2020). Here we examined which clusters of cells expressed Brachyury in relation to the morphogenesis and the identity of the ventral organizer. Our results showed that cells of all three endoderm clusters expressed Brachyury in blastulae. Based on expression profiles of genes involved in the gene regulatory networks (GRNs) of sea urchin embryos, the three clusters are distinguishable, two likely derived from the Veg2 tier and one from the Veg1 tier. On the other hand, of the four oral-ectoderm clusters, cells of two clusters expressed Brachyury at the gastrula stage and genes that are responsible for the ventral organizer at the late blastula stage, but the other two clusters did not. At a single-cell level, most cells of the two oral-ectoderm clusters expressed organizer-related genes, nearly a half of which coincidently expressed Brachyury. This suggests that the ventral organizer contains Brachyury-positive cells which invaginate to form the stomodeum. This scRNA-seq study therefore highlights significant roles of Brachyury-expressing cells in body-plan formation of early sea urchin embryos, though cellular and molecular mechanisms for how Brachyury functions in these processes remain to be elucidated in future studies.

Functional analysis of the eTM-miR171-SCL6 module regulating somatic embryogenesis in Lilium pumilum DC. Fisch.

Somatic embryogenesis (SE) is of great significance in Lilium bulb production, germplasm preservation, and genetic improvement. miRNAs are important regulators of plant growth and development at the transcriptional level. Previous research by our group has shown that lpu-miR171 and its target gene SCARECROW-LIKE 6 (SCL6) play an important regulatory role in lily SE, and we predicted and identified that endogenous target mimics (eTMs) can regulate lpu-miR171. However, the associated mechanism and internal regulatory network are not yet clear. In the present study, lpu-miR171 was used as an entry point to explore the regulatory network between its upstream eTMs and its downstream target gene LpSCL6, as well as to identify the mechanism of this regulatory network in Lilium SE. Tobacco transient transformation confirmed that miRNA171 significantly inhibited the expression of LpSCL6. On this basis, the Lilium stable genetic transformation system was used to demonstrate that silencing lpu-miR171a and lpu-miR171b and overexpressing LpSCL6-II and LpSCL6-I promoted starch accumulation in calli and the expression of key cell cycle genes, thus providing energy to meet preconditions for SE and accelerate the formation and development of Lilium somatic embryos. LpSCL6-II and LpSCL6-I are nuclear proteins with self-activation activity in yeast cells. In addition, we confirmed in Lilium that lpu-eTM171 is the eTM of lpu-miR171 that binds lpu-miR171 to prevent cleavage of the target gene LpSCL6, thereby promoting SE. Therefore, the present study established a new mechanism whereby the eTM-miR171-SCL6 module regulates SE in Lilium pumilum and provides new insights clarifying the mechanism of SE.

Building an embryo: An auxin gene toolkit for zygotic and somatic embryogenesis in Brazilian pine

Many studies in the model species Arabidopsis thaliana characterized genes involved in embryo formation. However, much remains to be learned about the portfolio of genes that are involved in signal transduction and transcriptional regulation during plant embryo development in other species, particularly in an evolutionary context, especially considering that some genes involved in embryo patterning are not exclusive of land plants. This study, used a combination of domain architecture phylostratigraphy and phylogenetic reconstruction to investigate the evolutionary history of embryo patterning and auxin metabolism (EPAM) genes in Viridiplantae. This approach shed light on the co-optation of auxin metabolism and other molecular mechanisms that contributed to the radiation of land plants, and specifically to embryo formation. These results have potential to assist conservation programs, by directing the development of tools for obtaining somatic embryos. In this context, we employed this methodology with critically endangered and non-model species Araucaria angustifolia, the Brazilian pine, which is current focus of conservation efforts using somatic embryogenesis. So far, this approach had little success since somatic embryos fail to completely develop. By profiling the expression of genes that we identified as necessary for the emergence of land-plant embryos, we found striking differences between zygotic and somatic embryos that might explain the developmental arrest and be used to improve A. angustifolia somatic culture.