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PubMed2001

Differentiation strategies of alternative splicing variants mRNA estrogen receptor-alpha.

From the clinical point of view recognizing the concentrations and type profile of isoforms could be of significant practical importance. The aim of the study is designed QRT-PCR reaction to assess profile of mRNA ER-alpha and their isoforms. Theoretical part of the study was made according to computer program and available Genbank database. To detect a isoform one of the primer was designed to hybridize within exon-border linked in alternative splicing. The study presents the differentiation strategies in isoforms coming about as the alternative splicing result. Designed oligonucleotide probes and primers allow to distinguish mRNA isoforms of ER-alpha and their quantification in assessed tissues.

Alternative Splicing↗

PubMed2001

Quantitative analysis of estrogen receptor-alpha and -beta and exon 5 splicing variant mRNA in endometrial hyperplasia in perimenopausal women.

We used real-time reverse transcription-polymerase chain reaction (QRT-PCR) to detect wild-type ERalpha and ERbeta and their isoforms as well as exon 5 splicing variant mRNA ERalpha in various types of endometrial hyperplasia in perimenopausal women. Our data suggest that the exon 2 and 7 deletion ERalpha mRNA isoforms are present in endometrial tissue undergoing various hyperplastic states. Wild type ERbeta mRNA and their isoforms 2, 4, 5, 6 were observed in each type of assessed endometrial specimens, and ERalpha/delta5 was found only in hyperplasia complex group.

Adult↗

PubMed2002

[Time-dependent effects of interleukin-8 gene expression in endothelial cells exposed on fluid shear stress].

Fluid shear stress plays an important role in vascular biology. In vivo, endothelial cells are continuously exposed to mechanical shear stress generated by the flowing blood. Previous studies have identified the exposure of vascular endothelial cells to fluid mechanical forces can modulate the expression of many genes involved in vascular physiology and pathophysiology. To investigate the role of fluid shear stress on IL-8 expression in human umbilical vein endothelial cells (HUVECs), we employed quantitative reversal transcription-polymerase chain reaction (qRT-PCR) to assay the expression of IL-8 mRNA. Here we show that IL-8 mRNA did not express in HUVECs untreated with fluid shear stress. IL-8 mRNA expression increased when HUVECs exposed to fluid shear stress for 1 h, and it reached the summit when HUVECs exposed to fluid shear stress for 2 h. Then IL-8 expression gradually decreased at 3 h of stimulation by shear stress and remained at a constant level throughout the time course of the study. The increase of IL-8 expression by shear stress was time-dependent. The biphasic response of IL-8 gene expression was found in experiments in which the applied shear stress was 2.23 dyne/cm2, 4.20 dyne/cm2, or 6.08 dyne/cm2. IL-8 gene expression in response to shear stress was very similar to NF-kappa B in response to shear stress. The induction of IL-8 gene expression by fluid shear stress is probably due to the activation of NF-kappa B. This in vitro study demonstrates the expression of IL-8 gene can be regulated by shear stress. Fluid shear stress induces a biphasic response of human IL-8 gene expression in HUVECs. These considerations suggest that IL-8 expression induced by fluid shear stress in HUVECs may play an important role in the genesis and development of both inflammation and arterioatherosclerosis.

Cells, Cultured↗

PubMed2002

[Force-dependent effects of interleukin-8 gene expression in endothelial cells exposed on fluid shear stress].

Fluid shear stress plays an important role in many physiological and pathophysiological processes of the cardiovascular system. Previous studies have identified the exposure of vascular endothelial cells to fluid mechanical forces can modulate the expression of many genes, including IL-8 gene expression, and IL-8 expression induced by fluid shear stress was time-dependent. To investigate the role of intensity of fluid shear stress on IL-8 expression in human umbilical vein endothelial cells (HUVECs), we had HUVECs exposed to shear stress 2.23, 4.20, 6.08, 8.19, 9.67, 12.15, 14.40, 16.87, and 19.29 dyne/cm2 respectively and employed quantitative reversal transcription-polymerase chain reaction (qRT-PCR) to assay the expression of IL-8 mRNA. The results show that IL-8 mRNA did not express in HUVECs untreated with fluid shear stress. The IL-8 mRNA expression by shear stress was force intensity-dependent. After HUVECs exposed to low fluid shear stress (2.23 dyne/cm2) for 1 h or 2 h, IL-8 mRNA expression increased near 68 or 52 times as that of HUVECs exposed to high fluid shear stress (19.29 dyne/cm2). The linear regression equations between IL-8 mRNA expression (log (copies), y) and shear stress (dyne/cm2, x) are: y = 7.57 - 0.11x, r = 0.97 (for 1 h); y = 7.92 - 0.10x, r = 0.96 (for 2 h). This in vitro study demonstrates the expression of IL-8 gene can be regulated by fluid shear stress. The low shear stress could induces much more expression of IL-8 mRNA, which plays probably an important role in the pathogenesis of inflammation and arteroatherosclerosis.

Cells, Cultured↗

PubMedDate not supplied

Induction of Epstein-Barr virus (EBV) reactivation in Raji cells by doxorubicin and cisplatin.

BACKGROUND: Epstein-Barr virus (EBV) can be activated in B-lymphoid cells to enter the lytic cycle by various kinds of stimuli, including 12-O-tetradecanoylphorbol-1 3-acetate (TPA), butyric acid, calcium ionophore A23187, transforming growth factor-beta and anti-immunoglobulin crosslinking. EBV reactivation has been clinically observed in patients receiving systemic chemotherapy. This study sought in vitro evidence to suggest whether anticancer drugs may directly contribute to the EBV reactivation. MATERIALS AND METHODS: Raji cells, an EBV-containing Burkitt's lymphoma cell line, were used as the experimental model. TPA served as a positive control for chemical induction of EBV reactivation. Expression of the BZLF1 transcript of EBV and its encoded protein, ZEBRA, were examined by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and flow cytometry, respectively. Transactivation activity of ZEBRA was further assessed by a luciferase reporter assay of EBV DR-promoter activity and a flow cytometry assay assessing the endogenous expression of EA-D (BMRFl). RESULTS: Doxorubicin and cisplatin, two commonly used anticancer agents, induced a dose-dependent up-regulation of BZLF1 mRNA and ZEBRA protein. The luciferase reporter activity and the expression of endogenous EA-D protein, also increased by doxorubicin and cisplatin, indicated an up-regulation of the transactivating activity of ZEBRA. CONCLUSION: These data indicate that cytotoxic anticancer drugs may up-regulate the expression and the transactivating activity of BZLF1, and suggest that systemic chemotherapy may be a risk factor for EBV reactivation in patients with EBV-associated malignancies.

Antigens, Viral↗

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↗

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↗

PubMed2025

Genome-wide identification, characterization, and expression pattern analysis of the glyoxalase gene family in Phyllostachys pubescens during abiotic stresses.

BACKGROUND: The glyoxalase pathway comprising of three enzymes i.e., glyoxalase I (GLYI), glyoxalase II (GLYII), and glyoxalase III (GLYIII), which play vital role in mitigating abiotic stresses by detoxifying the stress induced cytotoxic metabolite methylglyoxal (MG). Phyllostachys pubescens an ecologically and economically important forest species, plays vital roles in carbon sequestration and climate change mitigation. A genome-wide study was conducted to identify and characterize GLYI, GLYII, and unique DJ-1/GLYIII gene candidates in P. pubescens. The identified members were evaluated based on phylogenetic analysis, gene structure, chromosomal distribution, gene duplication, presence of conserved domain(s) and cis regulatory region. RESULTS: A total of 19 GLYI, 18 GLYII, and 15 GLYIII members were identified, each featuring characteristic domains: glyoxalase, metallo-β-lactamase, and DJ-1/PfpI, respectively. The presence of different cis-elements in the promoter region of the glyoxalase genes gives insights into their role and regulation under hormonal response, developmental processes and stress adaptation. Besides this, stress responsive transcription factors binding sites also dominated the promoter regions of glyoxalase genes. Expression analysis of various glyoxalase genes demonstrated significant variability under different stress conditions, underscoring their potential roles in stress modulation. Significant upregulation of all of the PhGLYI, PhGLYII, and PhGLYIII were observed under cold, drought, heavy metal and salinity stress suggesting their involvement in oxidative stress management, osmotic regulation and remodelling cellular redox homeostasis. Among the glyoxalase genes, PhGLYI-15, PhGLYII-9, and PhGLYIII-3 showed consistent upregulation under various abiotic stresses. CONCLUSIONS: Our findings reveal that glyoxalase genes crucially contribute towards the improvement of cellular osmotic potential in moso bamboo under different abiotic stresses. This study enhances our understanding of glyoxalase genes' evolution and functional roles in plants and opens new avenues for developing stress resilient crop varieties for sustainable agriculture.

Lactoylglutathione Lyase↗