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PubMed2003

[Expression patterns in isoforms of vascular endothelial growth-factors in tissue samples of vulval cancer T1 N2M0 stage].

OBJECTIVES: The aim of the study was estimating the expression of isoforms of mRNA vascular endothelial growth factor: VEGF121, VEGF145, VEGF165, VEGF183, VEGF189, VEGF206, in tissue samples of vulvar cancer, normal and inguinal lymph nodes metastases. MATERIALS AND METHODS: The material for this investigation of expression of mRNA selected genes was 4 tissue specimens from women with vulvar cancer T1N2M0. The method used for quantitative determination of the numbers of mRNA copies of selected genes was the QRT-PCR and QPCR technique, with the use of an ABI PRISM 7700 (TaqMan) sequence detector. The nucleotide sequence for starters and probes for quantitative RT-PCR was designed with the use of the Primer Express Version 1.0 ABI PRISM software. RESULTS: Expression of all mRNA isoforms VEGF was found in the samples. The number of mRNA copies obtained in 1 microgram total mRNA extract of the genes examined in tissue sections with various pathomorphology were clearly differentiated. CONCLUSIONS: Expression profile of aforementioned genes indicated a definitive relationship between vascular endothelial growth factor isoform and vulvar cancer. These findings provide evidence that the expression pattern of mRNA VEGF isoform should find broad applications in clinical and research settings.

Endothelial Growth Factors↗

PubMed2002

Transcriptional analysis of tumor-specific T-cell responses in cancer patients.

Over the last decade, tumor immunology in general and tumor immunotherapy in particular have made important progress. Thanks to the discovery of tumor-associated antigens (TAA), and the consequent development of active-specific immunization protocols, TAA-specific T-cell responses can now be induced reproducibly in cancer patients. However, clinical responses directly ascribable to TAA-specific T cells occur only occasionally. It is not clear why TAA-specific T cells do not eliminate tumor cells in vivo. This paradoxical coexistence of antigen-bearing tumor cells and antigen-specific T cells constitutes a critical point of current investigation. In recent years, protein-ligand interaction-based methods aimed at the identification and characterization of T cells using antibodies (cell surface phenotyping, intracellular and secreted cytokine detection), or HLA-peptide multimers, have significantly improved the analysis of TAA-specific T cells. These methods, however, seem to have reached the limit of their usefulness. Transcriptional analysis may add sensitivity and resolution and may provide global pictures of the multifactorial requirements for an efficient immune response against tumors. In this review, we describe the use of molecular genetic methods, such as real time qRT-PCR, cDNA microarrays, and TCR repertoire analysis. In addition, we describe techniques for high-fidelity messenger RNA amplification that allow high-throughput analysis of samples obtained from minimal sources, such as HLA/peptide tetramer sorted antigen-specific T cells, laser capture dissection, or fine needle aspirates. Recent work discussed in this review summarizes the complementarity of transcriptional analysis as an essential tool that, in addition to conventional methods, may deepen and broaden the characterization of tumor-specific T cells.

Gene Expression Profiling↗

PubMed2003

Selectively reduced expression of synaptic plasticity-related genes in amyloid precursor protein + presenilin-1 transgenic mice.

A critical question in Alzheimer's disease (AD) research is the cause of memory loss that leads to dementia. The amyloid precursor protein + presenilin-1 (APP+PS1) transgenic mouse is a model for amyloid deposition, and like AD, the mice develop memory deficits as amyloid deposits accumulate. We profiled gene expression in these transgenic mice by microarray and quantitative RT-PCR (qRT-PCR). At the age when these animals developed cognitive dysfunction, they had reduced mRNA expression of several genes essential for long-term potentiation and memory formation (Arc, Zif268, NR2B, GluR1, Homer-1a, Nur77/TR3). These changes appeared to be related to amyloid deposition, because mRNA expression was unchanged in the regions that did not accumulate amyloid. Transgene expression was similar in both amyloid-containing and amyloid-free regions of the brain. Interestingly, these changes occurred without apparent changes in synaptic structure, because a number of presynaptic marker mRNAs (growth-associated protein-43, synapsin, synaptophysin, synaptopodin, synaptotagmin, syntaxin) remained stable. Additionally, a number of genes related to inflammation were elevated in transgenic mice, primarily in the regions containing amyloid. In AD cortical tissue, the same memory-associated genes were downregulated. However, all synaptic and neuronal transcripts were reduced, implying that the loss of neurons and synapses contributed to these changes. We conclude that reduced expression of selected genes associated with memory consolidation are linked to memory loss in both circumstances. This suggests that the memory loss in APP+PS1 transgenic mice may model the early memory dysfunction in AD before the degeneration of synapses and neurons.

Aged↗

PubMed2003

[Differential expression of the epithelial membrane protein 1 of laryngeal carcinoma].

OBJECTIVE: In order to explore pathogenic mechanism of laryngeal carcinoma, the involved genes were identified in larynx carcinogenesis by comparing the gene expression profile in matched primary normal epithelial cells and primary laryngeal carcinoma cells from the same patients. METHODS: A cDNA microarray analysis consisting of 11,431 human genes revealed significant changes in the expression of 35 genes, with 8 genes being up-regulated and 27 being down-regulated. The epithelial membrane protein-1 (EMP-1) is one of the down-regulated genes. EMP-1 expression in various kinds of laryngeal carcinoma was determined by semi-quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). RESULTS: The EMP-1 mRNA levels in all laryngeal carcinoma cells was significantly lower than that in the matched primary normal epithelial cells (P < 0.05) and were not correlated to the stage and differentiation of laryngeal carcinoma (P > 0.05). CONCLUSIONS: The EMP-1 expression was correlated to larynx carcinogenesis and may be helpful to elucidate the pathogenic mechanism in laryngeal carcinoma.

Adult↗

PubMed2003

[Evaluation of wild-type estrogen receptor beta and its isoforms: ER-beta/delta 5/6 i ER-beta/delta 6 in endometrial adenocarcinoma].

Estrogen target genes play essential role among pathogenetic and prognostic factors of endometrial adenocarcinoma; and among them estrogen receptor genes. Although prevailing estrogen receptor type is ER-alpha in endometrium, transduction of signal carried by estrogens can be also mediate by ER-beta, which biological meaning relies on modulation of ER-alpha transcriptional activity. There is hypothesized that ER-beta can keep under control mitogenic activity mediating by ER-alpha. The aim of the study was qualitative and quantitative analysis of wtER-beta and ER-beta/delta 5-6, ER-beta/delta 6 isoforms in endometrial adenocarcinoma. Material used in the study included specimens obtained from 27 female: group I--normal endometrium (n = 12), group II--endometrial adenocarcinoma (n = 15). RNA was extracted from the analyzed material using phenol-chloroform method by Total RNA Prep Plus. Qualitative analysis was performed basing on RT-PCR reaction and polyacrylamide gel electrophoreses. For all RNA extracts of the samples examined RT-PCR was performed with a sequence detector ABIPRISMTM7700--Perkin-Elmer Applied Biosystems (RT: 600 C-30 min; PCR: 950 C-5 min, 40 cycles: 950 C-30s, 600 C-1 min, and 720 C-10 min) in order to determine the number of RNA copies (which corresponds to the number of cDNA copies) for wtER-beta and ER-beta/delta 5-6 and ER-beta/delta 6. Specificity of QRT-PCR reaction was determined by delimitation of melting temperature of all examined amplimers (ABI PRISM7000, Qu-antiTect SYBR Green RT-PCR Kit) and by sequence analysis using ABI PRISM377 DNA Sequencer. Wild-type of estrogen receptor beta and ER-beta/delta 5-6, ER-beta/delta 6 isoforms--coming into alternative splicing of mRNA, presented both, in proliferative endometrium and endometrial adenocarcinoma. Copy number of wtER-beta mRNA was significantly (p < 0.05, Mann-Whitney U test) lower in comparison with proliferative endometrium and correlated with decrease of ER-beta/delta 6 mRNA copy number (r = 0.85; p < 0.05). Preliminary results confirming decrease of wtER-beta, ER-beta/delta 5/6 and ER-beta/delta 6 mRNA copy number in endometrial adenocarcinoma can show their relationship with high risk of carcinogenesis.

Adenocarcinoma↗

PubMed2004

Expression of TGF beta1 genes and their receptor types I, II, and III in low- and high-grade malignancy non-Hodgkin's lymphomas.

BACKGROUND: Transforming growth factor beta (TGF ) is involved in a variety of important cellular functions. The lack of TGF -dependent cell-growth control might be related to oncogenesis, as it has been shown in lung, breast, and colon carcinomas. Current observations have revealed that TGF is rather an inhibiting, not a stimulating, factor as far as malignant tumor development is concerned. Recently, however, there has been a growing number of reports on increased expression of TGF genes in certain tumors. In patients with a diagnosis of non-small-cell lung carcinoma, the tumors expressing high levels of TGF were the ones that had a higher proliferation and metastasis capability, whereas more promising were the cases with lower levels of TGF expression. MATERIAL/METHODS: A pilot study of 14 patients was conducted comprising 8 patients with a low-grade lymphoma and 6 patients with a high-grade lymphoma. The QRT-PCR method was employed to assess the activity of TGF 1 and of its receptor types I, II, and III. RESULTS: The expression values for TGF 1 and its receptors I, II, and III were twice as high in the group of patients with a diagnosis of high-grade lymphomas as in the group of patients diagnosed with low-grade lymphomas. CONCLUSIONS: Results showed a clear difference in TGF 1 expression in patients with NHL depending on the subtype of the lymphoma, suggesting its significant role in the pathomechanism of this group of malignant diseases as well as its potential value as a prognostic factor.

Activin Receptors, Type I↗

PubMed2004

[Cell membrane phospholipid variation and protein kinase C expression effects on hepatic metastasis of large intestinal carcinoma].

OBJECTIVE: To study cell membrane phospholipid variation and protein kinase C (PKC) isoenzyme expression and their effects on hepatic metastasis of large intestinal carcinoma. METHODS: High function liquid chromatography was used to separate and detect cell membrane phospholipids of phosphatidylinosital (PI), phosphatidylserine (PS), phosphatidylethanolamine (PE) and phosphatidylcholine (PC) in primary foci, paratumor intestine mucosa and hepatic metastasis of large intestinal carcinomas. And mRNA expression levels of PKC-alpha, -beta II, -delta, -epsilon, -lambda, -zeta isoenzymes were detected using QRT-PCR technique. RESULTS: Fifty-eight cases of colorectal cancer were examined. CONTENTS: of PI, PC and PE in primary foci and hepatic metastasis were higher than those in paratumor mucosa. PE content in hepatic metastasis was much higher than that in primary foci (t = 98.88, P < 0.01). But PI and PC contents had no significant differences between primary and hepatic metastasis (t = 1.73, 1.36, P > 0.05). PKC-beta II, -delta, -epsilon, -lambda, -zeta expression were enhanced in primary foci and hepatic metastasis, but PKC-alpha level decreased in comparison with paratumor mucosa. And PKC-delta, -epsilon, -lambda, -zeta levels in hepatic metastasis were higher than those in primary foci (t = 4.31, P < 0.05). PI and PC had positive correlations with PKC-beta II expression. PE had positive correlations with PKC-delta, -epsilon, -lambda, -zeta, but a negative correlation with PKC-alpha. CONCLUSIONS: The increases of PI and PC and PKC-alpha/PKC-beta II ratio change are related with colorectal cancer genesis. High content of PE and enhanced expression of PKC-delta, -epsilon, -lambda, -zeta isoenzymes and decreased PKC-alpha level improved hepatic metastasis of large intestinal carcinoma.

Adult↗

PubMed2004

Effects of cell membrane phospholipid level and protein kinase C isoenzyme expression on hepatic metastasis of colorectal carcinoma.

BACKGROUND: The molecular mechanism of hepatic metastasis of colorectal cancer is not well understood. The aim of this study was to assess the relations between phospholipid contents of cellular membrane and isoenzyme expression of protein kinase C (PKC) and their effects on hepatic metastasis of colorectal cancer. METHODS: High performance liquid chromatography was used to detect contents of cell membrane phospholipids: phosphatidylinosital (PI), phosphatidylserine (PS), phosphatidylethanolamine (PE) and phosphatidylcholine (PC) in primary foci, paratumor mucosa and hepatic metastatic foci in patients with colorectal carcinoma. The mRNA expression levels of PKC-alpha, -betaII, -delta, -epsilon, -lambda, -zeta isoenzymes were detected with the QRT-PCR technique. RESULTS: The levels of PI, PC and PE in primary foci and hepatic metastatic foci were higher than those in paratumor mucosa. The level of PE in hepatic metastatic foci was much higher than that in primary foci (t=98.88, P<0.01); but the levels of PI and PC were not significantly different between primary foci and hepatic metastatic foci (t=1.73, 1.36, P>0.05). The expression levels of PKC-betaII, -delta, -epsilon, -lambda, -zeta were enhanced in primary foci and hepatic metastatic foci, but the level of PKC-alpha in primary foci was decreased as compared with that in paratumor mucosa. The levels of PKC-delta, -epsilon, -lambda, -zeta in hepatic metastatic foci were higher than those in primary foci. A positive correlation was observed between the expression levels of PI, PC and PKC-betaII and also between those of PE and PKC-delta, -epsilon, -lambda, -zeta. However, there was a close negative correlation between PE and PKC-alpha. CONCLUSION: Increased levels of PI and PC and decreased ratio of PKC-alpha to PKC-betaII are related to colorectal cancer genesis. Increased levels of PE, increased expression of PKC-delta, -epsilon, -lambda, -zeta isoenzymes and decreased level of PKC-alpha are related to hepatic metastasis in colorectal carcinoma.

Adult↗

PubMed2004

[Quantitative analysis of sigma genes expression in Mycobacterium tuberculosis cultures exposed to rifampicin and isoniazid].

The aim of the study was quantitative analysis of five genes encoding Mycobacterium tuberculosis sigma factors sigA, sigE, sigF, sigH, and sigI as well as the 85B reference gene known as the mycobacterial viability marker, in cultures exposed to rifampicin and isoniazid. The mRN levels were assessed using QRT-PCR technique, in the automated system of real time quantification with the ABI PRISM 7700 Sequence Detector System (TaqMan). The number of each analyzed gene transcript copies was expressed as a number of mRNA per 1 eg of isolated total RNA. In cultures exposed to the tested chemicals the number of 85B mRNA copies declined as compared to the controls (without tested chemicals). There was no detectable expression of sigA and sigI in the control cultures. Both, rifampicin and isoniazid induced expression of sigA and sigI genes. The sigE gene expression increased during exposure to isoniazid and decreased under rifampicin exposure conditions. The sigF mRNA was detected neither in the control culture, nor in cultures exposed to rifampicin or isoniazid. Both tested chemicals caused decrease of sigH expression.

Antibiotics, Antitubercular↗

PubMed2005

Recreational music-making modulates the human stress response: a preliminary individualized gene expression strategy.

BACKGROUND: A central component of the complex human biological stress response is the modulation of the neuro-endocrine-immune system with its intricate feedback loops that support homeostatic regulation. Well-documented marked gene expression variability among human and animal subjects coupled with sample collection timing and delayed effects, as well as a host of molecular detection challenges renders the quest for deciphering the human biological stress response challenging from many perspectives. MATERIAL/METHODS: A novel Recreational Music-Making (RMM) program was used in combination with a new strategy for peripheral blood gene expression analysis to assess individualized genomic stress induction signatures. The expression of 45 immune response-related genes was determined using a multiplex preamplification step prior to conventional quantitative Real Time Polymerase Chain Reaction (qRT-PCR) mRNA analysis to characterize the multidimensional biological impact of a 2-phase controlled stress induction/amelioration experimental protocol in 32 randomly assigned individuals. RESULTS: In subjects performing the RMM activity following a 1-hour stress induction protocol, 19 out of 45 markers demonstrated reversal with significant (P = 0.05) Pearson correlations in contrast to 6 out of 45 markers in the resting control group and 0 out of 45 in the ongoing stressor group. CONCLUSIONS: The resultant amelioration of stress-induced genomic expression supports the underlying premise that RMM warrants additional consideration as a rational choice within our armamentarium of stress reduction strategies. Modulation of individualized genomic stress induction signatures in peripheral blood presents a new opportunity for elucidating the dynamics of the human stress response.

Adolescent↗

PubMed2005

[Biological effects of nano-hydroxyapatite/polyamide 66 on the dental pulp cells].

OBJECTIVE: To investigate the biological effects of the new nano-hydroxyapatite/polyamide 66 biological composites (nHA-PA66) on the dental pulp cells. METHODS: After interaction with the nHA-PA66 eluate, the growth, proliferation, and function of the in vitro cultured human dental pulp cells were studied by cell culture technique, inverted phase-contrast microscope observation, MTT assay, flow cytometry, ALP activity assay and quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) analysis. RESULTS: The cultured pulp cells grew well and showed no morphological variation. Moreover, this material had no negative effects on the proliferation, cell cycle, ALP activity and the expression of dentin sialophosphoprotein (DSPP) mRNA of the pulp cells. CONCLUSION: As a new nano-biomaterial, nHA-PA66 has good biocompatibility to the pulp cells, but no obvious bioactivity.

Biocompatible Materials↗

PubMed2004

[Overexpression of thymosin beta4 in the cochlea of senescence-accelerated mouse].

OBJECTIVE: To develop gene expression profiles of young and old senescence-accelerated mouse (SAM) cochlea and identify genes responsible for aging-related hearing loss. METHODS: Gene micro-array slides containing 1101 mouse genes were hybridized to cDNA micro-arrays (Atlas Glass Array Mouse 1.0) that were synthesized using total RNAs from the cochlea of 2 mounts and 12 mounts mouse. Hybridization signals were visualized with cyanine-3 fluorescent reporter molecules, and the fluorescence intensities of the images were analyzed. Real-time quantitative reverse transcription PCR (qRT-PCR) was performed to validate the micro-array results. Immunofluorescence was used to identify the located region of the protein encoding by the candidate gene in the cochlea. RESULTS: Expression of a majority of the 1101 genes represented on the micro-array slides was not altered during aging; nonetheless, changes in the expression of 3 genes were detected between young and old mouse cochlea. RT-PCR results confirmed the changes in expression of thymosin beta4 of 3 genes examined. Through the using of immunofluorescence, it was shown that thymosin beta4 was primarily located in the tectorial membrane and the supporting cells of outer hair cell. CONCLUSIONS: Using commercially available slide micro-arrays, the results show that aging of the mouse cochlea is associated with changes in patterns of gene expression. This analysis suggests that thymosin beta4 may play a role in aging-related hearing loss. These studies lay the foundation for future studies defining the genetic basis of aging-related hearing loss.

Aging↗

PubMed2005

[Effect of ERK1/2 on low shear stress-induced expression of IL-8 mRNA in human endothelial cells].

Fluid shear stress plays an important role in many physiological and pathophysiological processes of the cardiovascular system. It modulates vascular function and structure via stimulating mechanosensitive endothelial cell signal events. Previous studies have identified that the exposure of vascular endothelial cells to fluid mechanical forces can modulate the expressions of many genes, including IL-8 gene. In order to gain an insight into the role of extracellular signal regulated kinase (ERK1/2) signal pathway in the expression of IL-8 mRNA in human umbilical vein endothelial cells (HUVECs) under the stimulation by low shear stress (4.20 dyne/cm2), we employed Western blot to measure phosphorylation of ERK1/2 and used quantitative reversal transcription-polymerase chain reaction (qRT-PCR) to assay the expression of IL-8 mRNA. The results showed: (1) Shear stress could activate ERK1/2 with a rapid, biphasic time course (maximum by 10 min and basal by 2 h); the treatment of HUVECs with Genistein (a highly specific inhibitor of tyrosine protein kinase, TPK) or PD98059 (the inhibitor of mitogen-activated protein/extracellular signal regulated kinase kinase, MEK) culd prevent shear-dependent activation of ERK1/2; (2) When treated with Genistein or PD98059, significant inhibition of IL-8 mRNA expression induced by low shear stress was observed in HUVECs. This in vitro study demonstrates that ERK1/2 plays an important role in IL-8 mRNA expression induced by low shear stress.

Cells, Cultured↗

PubMed2005

Effect of mosquito midgut trypsin activity on dengue-2 virus infection and dissemination in Aedes aegypti.

The effect of mosquito midgut trypsins in dengue serotype 2 flavivirus (DENV-2) infectivity to Aedes aegypti was studied. Addition of soybean trypsin inhibitor (STI) in a DENV-2 infectious blood meal resulted in a 91-97% decrease in midgut DENV-2 RNA copies (qRT-PCR analysis). STI treatment also resulted in slower DENV-2 replication in the midgut, less DENV-2 E protein expression, and decreased dissemination to the thorax and the head. A second uninfected blood meal, 7 days after the STI-treated infectious meal, significantly increased DENV-2 replication in the midgut and recovered oogenesis, suggesting that the lower viral infection caused by STI was in part due to a nutritional effect. Mosquitoes fed DENV-2 digested in vitro with bovine trypsin (before STI addition) exhibited a transient increase in midgut DENV-2 4 days postinfection. Blood digestion and possibly DENV-2 proteolytic processing, mediated by midgut trypsins, influence the rate of DENV-2 infection, replication, and dissemination in Ae. aegypti.

Aedes↗

PubMed2005

[Biological effects of nano-hydroxyapatite/polyamide 66 on the osteoblast].

OBJECTIVE: To evaluate the biological effects of nano-hydroxyapatite/polyamide 66(nHA-PA66) on the growth and activity of osteoblast. METHODS: MTT assay was used to determine the growth of osteoblast, enzymatic measure was used to determine the activity of ALP and quantitative RT-PCR (QRT-PCR) to evaluate the changes of osteoclacin mRNA expression in osteoblasts treated by DMEM eluate of nHA-PA66. RESULTS: Osteoblasts of different test groups demonstrated relative proliferation rate ranging from 98% - 106% without dose-dependent effect. The ALP activity and osteocalcin mRNA expression were similar in test and control groups (P > 0.05). CONCLUSION: nHA-PA66 has no negative effects on the osteoblast and its osteoblast-compatibility is proved.

Durapatite↗

PubMed2005

Galectin-9 as a regulator of cellular adhesion in human oral squamous cell carcinoma cell lines.

Oral squamous cell carcinomas (OSCCs), a major public health problem worldwide, are the most common neoplasms of the head and neck. The most important prognostic indicator for patients with OSCC is metastasis to cervical lymph nodes or distant organs. Galectin-9 is correlated with cellular adhesion and aggregation in melanoma cells. To investigate expression levels of galectin-9 mRNA and protein, we performed qRT-PCR and Western blot analyses on OSCC cell lines (Ca9-22, HSC-2, and HSC-3) and normal oral keratinocytes (NOKs). Galectin-9 mRNA and protein were commonly down-regulated in OSCC cell lines compared with NOKs. We further analyzed Ca9-22, which had the lowest expression of galectin-9. We then transfected the galectin-9 cDNA into Ca9-22 cells to examine whether overexpression of galectin-9 increases cellular adhesion in vitro. An adhesion assay using a fibronectin and collagen I coating plate revealed an increased cellular adherence ratio in overexpressed galectin-9 cells compared with nontransfected cells (p < 0.05). The data suggest that galectin-9 is correlated with oral cancer cell-matrix interactions and may therefore play an important role in the metastasis of OSCCs.

Blotting, Western↗

PubMed2025

Studying the Role of HOX Genes in Thrombocyte Development.

In our laboratory, we study thrombopoiesis and hemostasis using zebrafish as a model organism to unravel the mechanisms of differentiation and development of thrombocytes. We have shown in our earlier work that thrombocytes are functional equivalents of platelets and have transcriptional machinery similar to megakaryocytes. We recently found evidence that hox genes play a role in their development. We used piggyback gene knockdown and thrombocyte quantification assays to understand the influence of these ancient developmental regulators on thrombopoiesis. In this chapter, we describe methods used to discover these hox genes.

Animals↗

PubMed2026

Integrated metabolomics, transcriptional, and physicochemical analysis reveals key metabolites and genes associated with somatic embryogenesis in Phyllostachys pubescens.

Phyllostachys pubescens (Moso bamboo) is a significant perennial crop species that provides valuable nutritional and industrial uses, as well as carbon sequestration. Due to its remarkable growth rate, bamboo offers an ideal system for studying organogenesis, particularly in monocots. Somatic embryogenesis (SE) serves as a useful technique for crop breeding and improvement. SE in moso bamboo (Phyllostachys pubescens) remains challenging due to limited knowledge of its transcriptional and metabolomic reprogramming. To address this, we optimized callus initiation (MS + 18.1 µM 2,4-D + 8.5 µM picloram), callus proliferation (MS + 12.5 µM 2,4-D + 8.5 µM picloram), and somatic embryogenesis (MS + 1.1 µM 2,4-D + 3.3 µM metatopolin), using nodal segments as explants. UHPLC-Q-TOF-MS-based metabolite profiling revealed distinct biochemical trajectories across developmental stages of P. pubescens. NEC (non-embryogenic callus) was enriched in flavonoids, alkaloids, and saponins, while in-vitro shoots showed flavonoids and glycosides enrichment, and ex-vitro shoots showed high accumulation of glycosides and terpenoids. In contrast, EC (embryogenic callus) showed elevated levels of fatty acid derivatives (α-ESA, 26-Methyl Nigranoate), phytoalexins (Wyerone acid), sesquiterpene (Alpha-santalal, Beta-guaiene), flavonoid glycosides, and plant hormones (Cis-Zeatin, Gibberellin A45), indicating a metabolically active state supporting somatic embryogenesis. Similarly, genes and transcription factors controlling cell differentiation and embryogenesis were upregulated during SE. This study provides a comprehensive resource to facilitate future genomic and genetic investigations aimed at deciphering the molecular basis of organogenesis and advancing research on somatic embryogenesis in bamboo.

Plant Somatic Embryogenesis Techniques↗