Search Results (171)

Neurogenesis is considered the most robust form of plasticity in the adult brain. To better decipher this process, we evaluated the potential crosstalk of Kisspeptin and Endocannabinoid Systems (KPS and ECS, respectively) on hippocampal neurogenesis. Male adolescent rats were exposed to kisspeptin-10 (KP10) and the endocannabinoid anandamide (AEA) administered alone or in combination with the type 1 cannabinoid receptor (CB1R) antagonist SR141716A. The expression of Kiss1 and Kisspeptin receptor (Kiss1R) has been characterized for the first time in rat hippocampus together with the expression of the CB1R and the Transient Receptor Potential Vanilloid 1 ion channel receptor (TRPV1). Results show that both systems inhibit neurogenesis by reducing the extracellular signal-regulated kinase (ERK) signaling. Despite little differences in the expression of Kiss1R and CB1R, TRPV1 is enhanced by both KP10 and AEA treatments, suggesting TRPV1 as a common thread. KP10 administration reduces CB1R expression in the dentate gyrus, while AEA does not. KPS, unlike ECS, promotes the expression of estrogen receptor α (ER-α) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), also upregulating sirtuin 1 (SIRT1), brain-derived-neurotrophic factor (BDNF), and c-Jun. These findings suggest that the interaction between ECS and KPS could be involved in the fine-tuning of neurogenesis, highlighting a novel role for KPS. Full article

Functional gene expression is closely linked to an organism’s physiology and can be quantified using Real-Time Quantitative Reverse Transcription PCR (RT-qPCR). However, the stability of reference gene expression is not absolute, which may impact the accuracy of RT-qPCR results. In this study, we evaluated the suitability of nine genes including receptor for activated protein kinase c1 (rack1), ribosomal protein L6 (rpl6), ribosomal protein L9 (rpl9), ribosomal protein S2 (rps2), ribosomal protein S18 (rps18), ribosomal protein lateral stalk subunit P0 (rplp0), eukaryotic translation elongation factor 1β (eef1b), eukaryotic translation initiation factor 4a (eif4a), eukaryotic translation initiation factor 5a (eif5a) analyzed from RNA sequencing (RNA-Seq) data in addition to three genes including eukaryotic elongation factor 1α (eef1a), β-actin (actb), and glyceraldehyde 3-phosphate dehydrogenase (gapdh) selected from the literature to obtain the best internal controls in the RT-qPCR analysis of M. rosenbergii under overmating stress and natural aging. RefFinder was used to comprehensively evaluate the stability of the candidate reference genes. The initial results showed that three genes (eif5a, rps18, and rplp0) from the RNA-Seq data had relatively stable expression levels, which were more stable than those of the three commonly used reference genes. Eif5a and rps18 were the best combination for the RT-qPCR analysis of M. rosenbergii under overmating stress and aging. Further analysis indicated that eif5a might be the best reference gene for the study of M. rosenbergii. Full article

Background/Objectives: The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) enzyme, encoded by OsGAPDHC6, plays a crucial role in glycolysis while participating in various physiological and stress response pathways. Methods: In this study, the expression levels of the OsGAPDHC1 and OsGAPDHC6 genes were investigated over time by treating various abiotic stresses (ABA, PEG, NaCl, heat, and cold) in rice seedlings. Results: As a result, the expression levels of both genes in the ABA-treated group increased continuously for 0–6 h and then de-creased sharply from 12 h onwards. The mutational induction of the GAPDHC6 gene by the CRISPR/Cas9 system generated a stop codon through a 1 bp insertion into protein production. The knockout (KO) lines showed differences in seed length, seed width, and seed thickness compared to wild-type (WT) varieties. In addition, KO lines showed a lower germination rate, germination ability, and germination index of seeds under salt treatment compared to WT, and leaf damage due to 3,3′-diaminobenzidine (DAB) staining was very high due to malondialdehyde (MDA) accumulation. The KO line was lower regarding the expression level of stress-related genes compared to WT. Conclusions: Therefore, the OsGAPDHC6 gene is evaluated as a gene that can increase salt resistance in rice as it actively responds to salt stress in the early stages of growth, occurring from seed germination to just before the tilling stage. Full article

Heartworm disease is caused by Dirofilaria immitis, which mainly affects canids and felids. Adult D. immitis worms are located between the heart’s right ventricle and the pulmonary artery. These parasites produce an inflammatory and hypoxic process in the vascular endothelium. It has been demonstrated that D. immitis excretory/secretory antigens are able to stimulate the angiogenic process as a survival mechanism of D. immitis in the vascular endothelium, stimulating the proangiogenic pathway and related cellular processes. Our goal was to study the role of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and galectin (GAL) (proteins of D. immitis excretory/secretory antigens) plus vascular endothelial growth factor isoform A (VEGF-A) in the angiogenic process and their relationship with three cellular processes (cell proliferation, cell migration, and pseudocapillary formation) in an in vitro model of vascular endothelial cells. Cell viability and cytotoxicity were analyzed by live cell analysis and a commercial kit, respectively. VEGF-A, sVEGFR-2, VEGFR-1/sFlt, soluble endoglin, and membrane endoglin were analyzed by commercial ELISA kits. Cell proliferation, cell migration, and pseudocapillary formation were analyzed by MTT-based assay, the wound healing technique, and counting cell connections and cell clusters, respectively. rDiGAPDH+VEGF-A and rDiGAL+VEGF-A significantly increased the expression of sVEGFR-2, mEndoglin, and VEGF-A compared to cultures treated with only the proteins (rDiGAPDH and rDiGAL), VEGF-A, or unstimulated cultures. In addition, they also produced a significant increase in cell proliferation, cell migration, and pseudocapillary formation. Therefore, these proteins together with VEGF-A can activate the proangiogenic pathway and could be related to D. immitis survival in the circulatory system. Full article

Anthracnose is recognized as a significant agricultural disease. This study investigates the disease symptoms characterized by black dots on walnut fruits observed in the walnut orchards of Longnan City, Gansu Province, China, in June 2022. These symptoms resemble those of anthracnose reported in previous studies. A strain designated Ht-10 was initially isolated and identified as belonging to the Colletotrichum species based on its morphological features. Pathogenicity tests confirmed that this strain induced pronounced anthracnose symptoms in walnuts, consistent with those originally observed in the field. Subsequently, multilocus phylogenetic analysis, which included partial sequences of the internal transcribed spacer (ITS), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-tubulin2 (TUB2), and chitin synthase (CHS-1) genes of Ht-10, indicated that it most likely clustered with Colletotrichum fioriniae. The determination of biological characteristics revealed that the optimal temperature for the growth of Ht-10 was 25 °C in full light at a pH of 6, with soluble starch and yeast paste serving as the optimal carbon and nitrogen sources, respectively. To our knowledge, this is the first report of C. fioriniae as a causal agent of anthracnose in walnut fruits in China. Full article

Stem blight is a significant disease affecting blueberries worldwide, caused by various pathogens. This study investigated stem blight disease in Ji’an, Jilin Province, China. Fungi isolated from diseased stems were identified as Colletotrichum temperatum, Curvularia austriaca, and Diaporthe unshiuensis based on morphological characters and multi-locus phylogenetic analyses using the internal transcribed spacer (ITS) region, glyceraldehyde 3-phosphate dehydrogenase (gapdh), chitin synthase (chs), actin (act), β-tubulin (tub2), the translation elongation factor 1-alpha (tef1-α), calmodulin (cal), and histone 3 (his3) regions. Pathogenicity test was conducted on detached green blueberry shoots, all shoots inoculated by mycelium plugs presented necrotic lesions with dark brown margins, while the control (PDA plugs) group did not show any symptoms. Koch’s postulates were confirmed by re-isolating the inoculated pathogen from the disease symptoms. The study provides three new host–pathogen records of fungi associated with blueberry stem blight. Full article

Lucky bamboo (Dracaena sanderiana hort. ex. Mast. = Dracaena braunii) is a popular decorative plant in China. In March 2022, a severe outbreak of anthracnose disease occurred on the stems of lucky bamboo plants in a nursery garden in Nanjing, Jiangsu Province, China. Thirty-two fungal isolates were obtained from the infected stem tissues and were morphologically identified as Colletotrichum species. A multilocus phylogenetic analysis based on the internal transcribed spacer (ITS) region, the actin (ACT) gene, and the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene indicated the isolate FGZ-1 as Colletotrichum gloeosporioides (Penz.) Penz. and Sacc. The pathogenicity of isolate FGZ-1 was verified by inoculating mycelial plugs on stem segments and spraying spores on the whole one-year-old lucky bamboo plants. Koch’s postulates were fulfilled via the re-isolation of C. gloeosporioides from the diseased tissues. To the best of our knowledge, this is the first report of C. gloeosporioides causing anthracnose on lucky bamboo in China. The detection of C. gloeosporioides on lucky bamboo in China expands the range of Colletotrichum species that are associated with anthracnose in this popular ornamental plant. This study lays a solid foundation for future investigations into the pathogenic mechanisms of anthracnose on D. sanderiana and control strategies for this disease, such as biocontrol agents and the construction of resistant cultivars. Full article

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an enzyme widely involved in glycolysis in animal cells and in non-metabolic processes, including apoptosis and the regulation of gene expression. GAPDH is a ubiquitous protein that plays a pivotal role in plant metabolism and handling of stress responses. However, its function in plant stress resistance remains unknown. Identification and systematic analysis of the GAPDH family in Populus deltoides (P. deltoides) have not been performed. Bioinformatics methods were used to analyze the physicochemical characteristics, structural characteristics, phylogenetic relationships, gene structure, motif analysis, and expression of GAPDH gene family members in P. deltoides. We identified 12 GAPDH members in P. deltoides. Five types of PdGAPDH were identified: GAPA, GAPB, GAPC1, GAPC2, and GAPCp. PdGAPDH genes were differentially expressed in leaves, stems, and roots of 1-year-old poplar seedlings. PdGAPDH gene transcripts showed that PdGAPDH2 and PdGAPDH4 were highly expressed in the leaves. In the roots, seven genes—PdGAPDH01, PdGAPDH05, PdGAPDH06, PdGAPDH07, PdGAPDH08, PdGAPDH09, and PdGAPDH12—showed significantly high expression levels. PdGAPDH02, PdGAPDH03, PdGAPDH04, and PdGAPDH11 showed decreased expression under drought conditions and recovered after re-watering. These results lay the foundation for further studies on the drought stress mechanisms of P. deltoides. Full article

Background/Aim: Rosmarinus officinalis L. (R. officinalis) is an aromatic medicinal species with important nutraceutical potential, having rosmarinic acid (RA) as one of its main metabolites. The present study aims to evaluate the effects of an extract obtained from the leaves of this species and of its main metabolite in improving the streptozotocin-induced damage of hearts and aorta of diabetic rats. Methods: The leaves of the species were used to obtain a hydroethanolic extract, which was analyzed using the LC/MS method. Diabetes mellitus was induced by intraperitoneal streptozotocin administration in rats. After two weeks, oxidative stress parameters were evaluated from the heart and aorta homogenates. NOS3, AMPK, and adiponectin levels were quantified using ELISA tests, and thoracic aorta rings were isolated for contractility evaluation in the organ bath. Phospho-NF-κB, NRF2, HIF1 alfa, iNOS, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) quantification were performed using the Western blot technique. Results: Carnosic acid, together with rosmarinic acid, were proven to be the main metabolites identified in the composition of the tested extract. Administration of the extract and of RA improved the relaxation response to acetylcholine and the redox status, with the reduction in malondialdehyde (MDA), nitric oxide synthase 3 (NOS 3), AMP-activated protein kinase (AMPK), adiponectin, reduced (GSH) and oxidized glutathione (GSSG) levels, and superoxide dismutase (SOD) activity. RA significantly enhanced the expression of HIF 1α, NRF2, and pNFkB in the heart. Conclusions: Administration of the R. officinalis extract and of RA-alleviated oxidative stress, proving vascular and cardiac antioxidant properties in the hearts and aorta of diabetic rats. Full article

Antibiotic resistance has been and remains a major problem in our society. The main solution to this problem is to search and study the mechanisms of antibiotic action. Many groups of secondary metabolites, including antimicrobial ones, are produced by the Actinomycetota phylum. The actinobacterial strains isolated from habitats that have not been well studied are of great interest. Due to high resource competition, antibiotics are now considered a ‘trump card in the game of life’ due to their presence in natural substrates with limited nutrients. Potentially, strains isolated from such habitats can be producers of novel or poorly studied antibiotics. In the current research, we identified the strain Streptomyces sp. AP22 from the soils of the Akhshatyrsky Gorge, which is capable of producing pentalenolactone. This study describes the phenotypic and morphological characteristics of Streptomyces sp. AP22 and its biological activity. Pentalenolactone is a known inhibitor of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an important enzyme involved in glycolysis. We identified a previously unknown mutation in the gapA gene encoding glyceraldehyde-3-phosphate dehydrogenase that confers resistance to this antibiotic compound. This antibiotic is not used in clinical practice, so its application as a selectable marker will not lead to the creation of pathogens resistant to clinically relevant antibiotics. In this case, the selectable marker is based on a genetic construct containing the glyceraldehyde-3-phosphate dehydrogenase gene with a resistance mutation. The use of this selectable marker can be applied to various genetic and molecular techniques, such as cloning and transformation. This can help to facilitate genetic and molecular biology studies of strains resistant to standard selectable markers such as kanamycin or ampicillin. Full article

Anthracnose caused by Colletotrichum species is the most important disease of chayote (Sicyos edulis) in Brazil. The etiology of chayote anthracnose has been assigned to the species C. orbiculare, an important plant pathogenic fungus also reported as the causal agent of anthracnose in other cucurbits worldwide. However, there is no recent survey of the Colletotrichum species causing anthracnose in chayote in Brazil. In this study, Colletotrichum isolates associated with anthracnose on the fruit and leaves of chayote, from various producing regions in Brazil, were collected and identified. Haplotype analysis based on sequences of the β-tubulin genomic region (TUB2) of 44 Colletotrichum isolates was carried out as a first measure of genetic diversity. A subset of 22 isolates were sequenced using the partial sequences of actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and the rDNA ITS (ITS) region. Maximum likelihood analysis was performed using the concatenated sequences. The multilocus sequence analysis revealed four previously described species, Colletotrichum chrysophilum, C. menezesiae, C. plurivorum, and C. karsti, and two novel species, named C. cucurbitacearum and C. sicyi. All species were able to induce typical symptoms of anthracnose in chayote fruits but varied in their aggressiveness. The species C. menezesiae and C. sicyi were the most aggressive, while C. plurivorum was the least aggressive. The species C. orbiculare was not found to cause chayote anthracnose in Brazil. Full article

In this study, we carried out large-scale leaf spot symptom observation on vineyards in the Krasnodar Krai of Russia and determined their distribution. The incidence and severity of leaf spot were higher on the Euro-American grapevine hybrids (Bianka, Levokumskij, Avgustin, Moldova, Pervenets Magaracha, Dunavski lazur). A total of 433 isolates that belonged to the genus Alternaria were isolated from samples with leaf spot. Pathogenicity testing confirmed the ability of the representative isolates to cause necrosis on the grapevine. The isolates of Alternaria sp. were typed by the loci of internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (gapdh), Alternaria allergen a1 (Alt a1), β-tubulin (tub), and translation elongation factor (tef1). Isolates from grapevine causing leaf spot were shown to cluster with isolates of Alternaria. alternata (Fr.) Keissl. and Alternaria. arborescens E.G. Simmons species complex. Of the fungicides tested to inhibit Alternaria growth, the most effective were mixtures, such as pyrimethanil and fluopyram, cyprodinil and fludioxonil, and those that included difenoconazole. The results of the study expand our knowledge of the biodiversity of Alternaria sp. fungi and can be used to limit the spread of Alternaria leaf spot of the grapevine. Full article

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a major glycolytic enzyme that plays an important role in several cellular processes, including plant hormone signaling, plant development, and transcriptional regulation. In this study, we divided it into four groups through structural analysis of eight GAPDH genes identified in the rice genome. Among them, the expression level of five genes of cytosolic GAPDH was shown to be different for each organ. The mutation induction of the GAPDHC7 gene by the CRISPR/Cas9 system revealed that the 7 bp and 2 bp deletion, early end codon, was used in protein production. In addition, the selected mutants showed lower plant heights compared to the wild-type plants. To investigate the effect on carbohydrate metabolism, the expression of the genes of starch-branched enzyme I (SbeI), sucrose synthase (SS), and 3-phosphoglycer phosphokinase (PGK) increased the expression of the SBeI gene threefold in the knockout lines compared to the wild-type (WT) plant, while the expression of the SS and PGK genes decreased significantly. And the starch and soluble sugar content of the knockout lines increased by more than 60% compared to the WT plant. Also, the free amino acid content was significantly increased in the Gln and Asn contents of the knockout lines compared to the WT plants, while the contents of Gly and Ser were decreased. Our results suggest that OsGAPDHC7 has a great influence on energy metabolism, such as pre-harvested sprouting and amino acid content. Full article

Colletotrichum species are the most common cause of maize anthracnose, which often manifests as leaf spots. However, field observations often reveal symptoms similar to those caused by other leaf spot pathogens, such as Curvularia and Bipolaris. In this study, 99 isolates were identified using tissue separation and single-spore isolation techniques. As preliminary measures of species diversity, all 99 isolates were identified morphologically, and the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene sequences were phylogenetically analyzed. Furthermore, 48 representative strains were selected for molecular identification using multi-locus phylogenetic analyses based on five gene loci (ITS, TUB, ACT, GAPDH, and CAL). Finally, 10 species of Colletotrichum isolated from maize leaf spots were identified. Colletotrichum cliviicola was the most dominant species (24.2%), followed by C. fructicola (18.2%), C. karstii (16.1%), C. siamense (13.1%), C. boninense (7.1%), C. kahawae (7.1%), C. brevisporum (6.1%), C. truncatum (5.1%), C. gigasporum (2.0%), and C. gloeosporioides (1.0%). For the first time, pathogenicity tests revealed that C. cliviicola, C. fructicola, C. siamense, C. karstii, and C. truncatum are the causative agents of maize anthracnose. Additionally, C. boninense was identified as an endophytic fungus on healthy maize. In conclusion, this study is the first to identify the pathogen of maize anthracnose in Sichuan Province. It provides valuable insights for accurately diagnosing and managing maize anthracnose. Full article

Macadamia is an economically significant crop, with its kernel oil being abundant in monounsaturated fatty acids (MUFA). Analyzing the expression of genes related to MUFA biosynthesis is essential for understanding the complex regulatory networks in Macadamia. However, there are few reports on the identification of suitable reference genes for use as internal controls in this species. Consequently, selecting a reliable reference gene for gene expression studies under various conditions is critical. In this study, we evaluated the expression stability of 11 traditional housekeeping genes: α-tubulin (TUBa), β-tubulin (TUBb), malate dehydrogenase (MDH), 18S ribosomal RNA (18S), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), α-elongation factor 1 (EF1a), β-elongation factor 1 (EF1b), ubiquitin (UBQ), ubiquitin-conjugating enzyme (UBC), cyclophilin (CYP), and actin (ACT) under abiotic stresses, hormonal treatments and in variety of plant tissues using the online tool RefFinder, which integrates four commonly used software programs: ΔCt, geNorm (version 3.4), NormFinder (version 0953), and BestKeeper (version 1.0). A comprehensive expression stability ranking was established by integrating results from these four methods based on the geometric mean. The findings indicated that ACT was the most stable gene across all samples, including those subjected to cold stress, NaCl stress, PEG stress, ABA treatment, MeJA treatment, and both stem and leaf tissues. EF1b was identified as the most stable gene in GA treatment and heat stress samples, while UBC and CYP were ranked highest in ethrel treatment and root tissue samples, respectively. Finally, the reliability of these findings was further validated using the target gene SAD through qRT-PCR. In summary, this study evaluated and validated appropriate reference genes for qRT-PCR, which will facilitate future investigations into the molecular mechanisms in Macadamia. Full article