Cytogenetics, Typification, Molecular Phylogeny and Biogeography of Bentinckia (Arecoideae, Arecaceae), an Unplaced Indian Endemic Palm from Areceae.

Bentinckia is a genus of flowering plants which is an unplaced member of the tribe Areceae (Arecaceae). Two species are recognized in the genus, viz. B. condapanna Berry ex Roxb. from the Western Ghats, India, and B. nicobarica (Kurz) Becc. from the Nicobar Islands. This work constitutes taxonomic revision, cytogenetics, molecular phylogeny, and biogeography of the Indian endemic palm genus Bentinckia. The present study discusses the ecology, morphology, taxonomic history, distribution, conservation status, and uses of Bentinckia. A neotype was designated for the name B. condapanna. Cytogenetical studies revealed a new cytotype of B. condapanna representing 2n = 30 chromosomes. Although many phylogenetic reports of the tribe Areceae are available, the relationship within the tribe is still ambiguous. To resolve this, we carried out Bayesian Inference (BI) and Maximum Likelihood (ML) analysis using an appropriate combination of chloroplast and nuclear DNA regions. The same phylogeny was used to study the evolutionary history of Areceae. Phylogenetic analysis revealed that Bentinckia forms a clade with other unplaced members, Clinostigma and Cyrostachys, and together they show a sister relationship with the subtribe Arecinae. Biogeographic analysis shows Bentinckia might have originated in Eurasia and India.

Revised molecular phylogeny, global biogeography, and diversification of palms subfamily Coryphoideae (Arecaceae) based on low copy nuclear and plastid regions.

Coryphoideae are palmate-leaved palms from the family Arecaceae consisting of 46 genera representing 421 species. Although several phylogenetic analyses based on different genomic regions have been carried out on Coryphoideae, a fully resolved molecular phylogenetic tree has not been reported yet. To achieve this, we applied two phylogenetic reconstruction methods: Maximum Likelihood and Bayesian Inference, using amplified sampling by retrieving chloroplast and nuclear DNA sequences from NCBI and adding newly produced sequences from Indian accession into the dataset. The same dataset (chloroplast + nuclear DNA sequences) was used to estimate divergence times and the evolutionary history of Coryphoideae with a Bayesian uncorrelated, lognormal relaxed-clock approach and a Statistical Divergence-Vicariance Analysis method, respectively. The phylogenetic analyses based on a combined chloroplast and nuclear DNA sequence dataset showed well-resolved relationships within the subfamily. Both phylogenetic trees divide Coryphoideae into two main groups: CSPT (Crysophileae, Sabaleae, Phoeniceae, and Trachycarpeae) and the Syncarpous group. These main groups are segregated into eight tribes (Trachycarpeae, Phoeniceae, Sabaleae, Crysophileae, Borasseae, Corypheae, Caryoteae, and Chuniophoeniceae) and four subtribes (Rhapidine, Livistoninae, Hyphaeninae, and Lataniinae) with strong support-values. Most previously unresolved and doubtful relationships within tribes Trachycarpeae and Crysophilieae are now resolved and well-supported. The reconstructed phylogenetic trees support all previous systematic revisions of the subfamily. All Indian sampled species of Arenga, Bentinckia, Hyphaene, and Trachycarpus show close relation with their respective congeneric species. Molecular dating results and integration of biogeography suggest that Coryphoideae originated in Laurasia at ~95.12 Ma and then diverged into the tropical and subtropical regions of the whole world. This study offers the correct combination of nuclear and plastid regions to test the current and future systematic revisions.

Gene Expression Analyses of Mutant Flammulina velutipes (Enokitake Mushroom) with Clogging Phenomenon.

Regulation of proper gene expression is important for cellular and organismal survival, maintenance, and growth. Abnormal gene expression, even for a single critical gene, can thwart cellular integrity and normal physiology to cause diseases, aging, and death. Therefore, gene expression profiling serves as a powerful tool to understand the pathology of diseases and to cure them. In this study, the difference in gene expression in Flammulina velutipes was compared between the wild type (WT) mushroom and the mutant one with clogging phenomenon. Differentially expressed transcripts were screened to identify the candidate genes responsible for the mutant phenotype using the DNA microarray analysis. A total of 88 genes including 60 upregulated and 28 downregulated genes were validated using the real-time quantitative PCR analysis. In addition, proteomic differences between the WT and mutant mushroom were analyzed using two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF). Interestingly, the genes identified by these genomic and proteomic analyses were involved in stress response, translation, and energy/sugar metabolism, including HSP70, elongation factor 2, and pyruvate kinase. Together, our data suggest that the aberrant expression of these genes attributes to the mutant clogging phenotype. We propose that these genes can be targeted to foster normal growth in F. velutipes.

Construction and implementation of floating wetpark as effective constructed wetland for industrial textile wastewater treatment.

Fimbristylis dichotoma, Ipomoea aquatica, Pluchea tomentosa and their co-plantation (consortium FIP) autonomously degrade Orange 3R. Consortium FIP showed 84% removal of Orange 3R within 48 h, which is a higher dye elimination rate than individual plant systems. Oxidoreductase enzymes like tyrosinase (76%), varatryal alcohol oxidase (85%), lignin peroxidase (150%), riboflavin reductase (151%), laccase (171%), NADH-DCIP reductase (11%) and azo reductase (241%) were expressed in consortia FIP during Orange 3R degradation. UV-vis spectroscopy, enzyme activities, HPTLC, FTIR and GC-MS confirmed mineralization of Orange 3R into its metabolites. Microscopic investigation of root tissue revealed the harsh effect of dye on root tissues. Toxicity assessment on the HepG2 cell line demonstrated the toxic nature of Orange 3R, which gets reduced after phyto-treatment with consortia FIP. Floating wetpark of consortia FIP was found more efficient for the treatment of industrial textile waste and accomplished 87%, 86%, 75%, 49% and 46% removal of COD, BOD, color, TSS and TDS of effluent.

Overexpression of cathepsin S exacerbates lupus pathogenesis through upregulation TLR7 and IFN-α in transgenic mice.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that affects multiple organs. Recent studies suggest relevance between cysteine protease cathepsin S (CTSS) expression and SLE. To investigate the mechanism of CTSS in SLE, CTSS-overexpressing transgenic (TG) mice were generated, and induced lupus-like symptoms. Eight months later, the TG mice spontaneously developed typical SLE symptoms regardless of the inducement. Furthermore, we observed increased toll-like receptor 7 (TLR7) expression with increased monocyte and neutrophil populations in the TG mice. In conclusion, overexpression of CTSS in mice influences TLR7 expression, autoantibodies and IFN-α, which leads to an autoimmune reaction and exacerbates lupus-like symptoms.

20(S)-Ginsenoside Rh2 Suppresses Oral Cancer Cell Growth by Inhibiting the Src-Raf-ERK Signaling Pathway.

BACKGROUND: 20(S)-Ginsenoside Rh2 (G-Rh2) has demonstrated therapeutic effects in many types of cancers. We aimed to investigate the potential anticancer activity and underlying mechanisms of G-Rh2 in oral cancer cells. MATERIALS AND METHODS: The antigrowth effect of G-Rh2 in oral cancer cells was stimulated by cell proliferation, soft agar colony formation, and migration and invasion assay. The cell cycle and apoptosis were detected by flow cytometry. The underlying mechanism of G-Rh2 in oral cancer cells was explored by immunoblotting. RESULTS: G-Rh2 significantly inhibited oral cancer cell growth by inducing apoptosis and cell cycle G0/G1-phase arrest. G-Rh2 inhibited oral cancer cell migration and invasion through regulation of epithelial-mesenchymal transition (EMT)-related proteins. G-Rh2 inhibited the Src/Raf/ERK signaling pathway in YD10B and Ca9-22 cells. CONCLUSION: G-Rh2 exerted anticancer activity in vitro by inhibiting the Src/Raf/ERK signaling pathway in oral cancer. G-Rh2 is a potential therapeutic drug for oral cancer treatment.

Diversity and Plant Growth-Promoting Effects of Fungal Endophytes Isolated from Salt-Tolerant Plants.

Fungal endophytes are symbiotic microorganisms that are often found in asymptomatic plants. This study describes the genetic diversity of the fungal endophytes isolated from the roots of plants sampled from the west coast of Korea. Five halophytic plant species, Limonium tetragonum, Suaeda australis, Suaeda maritima, Suaeda glauca Bunge, and Phragmites australis, were collected from a salt marsh in Gochang and used to isolate and identify culturable, root-associated endophytic fungi. The fungal internal transcribed spacer (ITS) region ITS1-5.8S-ITS2 was used as the DNA barcode for the classification of these specimens. In total, 156 isolates of the fungal strains were identified and categorized into 23 genera and two phyla (Ascomycota and Basidiomycota), with Dothideomycetes and Sordariomycetes as the predominant classes. The genus Alternaria accounted for the largest number of strains, followed by Cladosporium and Fusarium. The highest diversity index was obtained from the endophytic fungal group associated with the plant P. australis. Waito-C rice seedlings were treated with the fungal culture filtrates to analyze their plant growth-promoting capacity. A bioassay of the Sm-3-7-5 fungal strain isolated from S. maritima confirmed that it had the highest plant growth-promoting capacity. Molecular identification of the Sm-3-7-5 strain revealed that it belongs to Alternaria alternata and is a producer of gibberellins. These findings provided a fundamental basis for understanding the symbiotic interactions between plants and fungi.

Therapeutic potential of the mitochondria-targeted antioxidant MitoQ in mitochondrial-ROS induced sensorineural hearing loss caused by Idh2 deficiency.

Mitochondrial NADP+-dependent isocitrate dehydrogenase 2 (IDH2) is a major NADPH-producing enzyme which is essential for maintaining the mitochondrial redox balance in cells. We sought to determine whether IDH2 deficiency induces mitochondrial dysfunction and modulates auditory function, and investigated the protective potential of an antioxidant agent against reactive oxygen species (ROS)-induced cochlear damage in Idh2 knockout (Idh2-/-) mice. Idh2 deficiency leads to damages to hair cells and spiral ganglion neurons (SGNs) in the cochlea and ultimately to apoptotic cell death and progressive sensorineural hearing loss in Idh2-/- mice. Loss of IDH2 activity led to decreased levels of NADPH and glutathione causing abnormal ROS accumulation and oxidative damage, which might trigger apoptosis signal in hair cells and SGNs in Idh2-/- mice. We performed ex vivo experiments to determine whether administration of mitochondria-targeted antioxidants might protect or induce recovery of cells from ROS-induced apoptosis in Idh2-deficient mouse cochlea. MitoQ almost completely neutralized the H2O2-induced ototoxicity, as the survival rate of Idh2-/- hair cells were restored to normal levels. In addition, the lack of IDH2 led to the accumulation of mitochondrial ROS and the depolarization of ΔΨm, resulting in hair cell loss. In the present study, we identified that IDH2 is indispensable for the functional maintenance and survival of hair cells and SGNs. Moreover, the hair cell degeneration caused by IDH2 deficiency can be prevented by MitoQ, which suggests that Idh2-/- mice could be a valuable animal model for evaluating the therapeutic effects of various antioxidant candidates to overcome ROS-induced hearing loss.

Diversity and Plant Growth Promoting Capacity of Endophytic Fungi Associated with Halophytic Plants from the West Coast of Korea.

Five halophytic plant species, Suaeda maritima, Limonium tetragonum, Suaeda australis, Phragmites australis, and Suaeda glauca Bunge, which are native to the Muan salt marsh of South Korea, were examined for fungal endophytes by sequencing the internal transcribed spacer (ITS) region containing ITS1, 5.8S rRNA, and ITS2. In total, 160 endophytic fungal strains were isolated and identified from the roots of the 5 plant species. Taxonomically, all 160 strains belonged to the phyla Ascomycota, Basidiomycota, and Zygomycota. The most dominant genus was Fusarium, followed by the genera Penicillium and Alternaria. Subsequently, using 5 statistical methods, the diversity indices of the endophytes were determined at genus level. Among these halophytic plants, P. australis was found to host the greatest diversity of endophytic fungi. Culture filtrates of endophytic fungi were treated to Waito-C rice seedlings for plant growth-promoting effects. The fungal strain Su-3-4-3 isolated from S. glauca Bunge provide the maximum plant length (20.1 cm) in comparison with wild-type Gibberella fujikuroi (19.6 cm). Consequently, chromatographic analysis of the culture filtrate of Su-3-4-3 showed the presence of physiologically active gibberellins, GA1 (0.465 ng/mL), GA3 (1.808 ng/mL) along with other physiologically inactive GA9 (0.054 ng/mL) and GA24 (0.044 ng/mL). The fungal isolate Su-3-4-3 was identified as Talaromyces pinophilus.

A1555G homoplasmic mutation from A1555G heteroplasmic mother with Pendred syndrome.

Hearing loss (HL) is genetically heterogeneous and can be caused by mutations in multiple gene lesions. Pendred syndrome, caused by mutation of SLC26A4, is one of the common causes of recessive syndromic profound HL. Mitochondrial mutation is another rare cause of genetic HL, resulting in late onset sensorineural HL. Recently, we evaluated a young woman representing bilateral progressive moderate HL with delayed language development, along with her family. Hearing test, temporal bone computed tomography, and genetic evaluation of GJB2, MT-RNR1, SLC26A4 gene mutations were performed on each family member. Her mother was prelingually deaf and displayed enlarged vestibular aqueduct (EVA) along with goiter. Interestingly, subject's mother showed both SLC26A4 mutation and mitochondrial A1555G heteroplasmic mutation at the same time. The sisters did not display EVA or goiter. Although the subject's older sister showed both prelingual deafness and mitochondrial A1555G heteroplasmy, her younger sister showed only A1555G homoplasmy, which suggests A1555G homoplasmy as the genetic cause of hearing loss. This is the first report of HL caused by mitochondrial A1555G homoplasmy from a mother with Pendred syndrome coexistent with A1555G heteroplasmy in the Korean population.

Culturable fungal endophytes isolated from the roots of coastal plants inhabiting korean East coast.

Twelve plant species were collected from the east coast of Korea to identify culturable endophytes present in their roots. The fungal internal transcribe spacer (ITS) region (ITS1-5.8SrRNA-ITS2) was used as a DNA barcode for identification of fungi. A total of 194 fungal strains were identified and categorized into 31 genera. The genus Penicillium accounted for the largest number of strains, followed by the genus Aspergillus. Furthermore, using 5 statistical methods, the diversity indices of the fungi were calculated at the genus level. After comprehensive evaluation, the endophytic fungal group from Phragmites australis ranked highest in diversity analyses. Several strains responsible for plant growth and survival (Penicillium citrinum, P. funiculosum, P. janthinellum, P. restrictum, and P. simplicissimum), were also identified. This study provides basic data on the sheds light on the symbiotic relationship between coastal plants and fungi.

Cadophora malorum Cs-8-1 as a new fungal strain producing gibberellins isolated from Calystegia soldanella.

Fourteen endophytic fungi with different colony morphologies were isolated from the roots of Calystegia soldanella. Endophytic fungi isolated from C. soldanella were identified by internal transcribed spacer (ITS) region. To verify plant growth promotion (PGP), culture filtrates of isolated endophytic fungi were treated in Waito-c rice (WR) and C. soldanella seedlings. Culture filtrates of Cs-8-1 fungal strain had advanced PGP activity. The presence of physiologically bioactive gibberellins (GA) GA(1) (1.213 ng ml(-1)), GA(3) (1.292 ng ml(-1)), GA(4) (3.6 ng ml(-1)), GA(7) (1.328 ng ml(-1)), other inactive GA(9) (0.796 ng ml(-1)) and GA(12) (0.417 ng ml(-1)), GA(20) (0.302 ng ml(-1)), GA(24) (1.351 ng ml(-1)), GA(34) (0.076 ng ml(-1)), and GA(53) (0.051 ng ml(-1)) in culture filtrates of Cs-8-1 fungal strain was detected. The Cs-8-1 fungal strain was confirmed as a producer of GAs. Molecular analysis of sequences showed high similarity of 99% to Cadophora malorum. Consequentially, the Cs-8-1 fungal strain was identified as a new C. malorum producing GAs.

Fungal diversity and plant growth promotion of endophytic fungi from six halophytes in Suncheon Bay.

Endophytic fungi were isolated from roots of six halophytes in Suncheon Bay. The endophytic fungi of 35 species isolated from halophytes were identified by internal transcribed spacer (ITS) containing the ITS1, 5.8s, and ITS2 regions. All fungal strains were analyzed to diversity at the genus level. Fungal culture filtrates (FCF) of endophytic fungi were treated to Waito-c rice (WR) seedling for plant growth-promoting verification. It was confirmed that fungal strain Sj-2-2 provided plant growth promotion (PGP) to WR seedling. Then, PGP of Suaeda japonica was confirmed by treating culture filtrate of Sj-2-2. As a result, it was verified that culture filtrate of Sj-2-2 had more advanced PGP than positive control when treated to S. japonica. The secondary metabolites involved in culture filtrate of Sj-2-2 were identified by HPLC and GC-MS SIM analysis. The presence of physiologically bioactive gibberellins (GAs) and other inactive GAs in culture filtrate of Sj-2-2 was detected. The molecular analysis of sequences of Sj-2-2 showed the similarity to Penicillium sp. of 99% homology. The PGP of Sj-2-2 as well as symbiosis between endophytic fungi and halophytes growing naturally in salt marsh was confirmed. Sj-2-2 was identified as a new fungal strain producing GAs by molecular analysis of sequences. Consequently, the Sj-2-2 fungal strain was named as Penicillium sp. Sj-2-2. In this study, the diversity of endophytic fungi isolated from roots of halophytes in salt marsh and the PGP of a new gibberellin-producing fungal strain were confirmed.

Mutualistic association of Paecilomyces formosus LHL10 offers thermotolerance to Cucumis sativus.

We investigated in this study the influence of an endophytic fungus, Paecilomyces formosus LHL10, on the thermotolerance of cucumber (Cucumis sativus) upon exposure to high (38°C) and low (8°C) temperature stresses. The results showed that endophyte-inoculated plants had significantly higher plant growth attributes under high-temperature stress. However, they were either low or insignificant in non-inoculated control and inoculated plants with 8°C treatments. Lower stress-promulgated water deficit and cellular membrane damage were observed in endophyte-treated plants after 38°C treatment than in control plants under 8°C stress. Total polyphenol, reduced glutathione, and lipid peroxidation activities were reduced in endophyte-associated plants after exposure to 38°C as compared with control and 8°C-treated plants. The concentration of saturated fatty acids (palmitic-C16:0; stearic-C18:0) was lower in endophyte-treated plants with or without low-temperature stress, but after 8°C treatment increased compared with controls. Unsaturated fatty acids (oleic-C18:1; linoleic-C18:2; linolenic-C18:3 acids) were similar at normal conditions; however, at 38°C, C18:2 and C18:3 were decreased, and C18:1 was increased in endophyte-treated plants compared with controls, while the inverse relationship was found at 8°C. Low levels of abscisic acid in P. formosus-associated plants after 38°C treatments revealed stress tolerance compared with control and 8°C-treated plants. In contrast, salicylic acid was pronounced in endophyte-treated plants after low-temperature stress as compared to other treatments. The results provide evidence that the response to P. formosus association was beneficial at normal growth temperature and had varying effects in response to temperature stress.

Overexpression of Jazf1 induces cardiac malformation through the upregulation of pro-apoptotic genes in mice.

The transcription factor Juxtaposed with another zinc finger gene 1 (JAZF1) is a zinc finger protein that binds to the nuclear orphan receptor TR4. Recent evidence indicates that TR4 receptor functions as both a positive and negative regulator of transcription, but the role of JAZF1 in transcriptional mechanisms has not been elucidated. Recently, the incidence rate of congenital heart malformations was reported to be significantly elevated in patients who had neurofibromatosis 1 (NF1) with chromosomal microdeletion syndrome. Furthermore, Joined to JAZF1 (SUZ12) is expressed at high levels in the hearts of adult patients with NF1 microdeletion syndrome. Therefore, we hypothesized that ectopic expression of JAZF1 may lead to cardiac malformations that deleteriously affect the survival of neonates and adults. We sought to elucidate the role of JAZF1 in cardiac development using a Jazf1-overexpressing (Jazf1-Tg) mouse model. In Jazf1-Tg mice, Jazf1 mRNA expression was significantly elevated in the heart. Jazf1-Tg mice also showed cardiac defects, such as high blood pressure, electrocardiogram abnormalities, apoptosis of cardiomyocytes, ventricular non-compaction, and mitochondrial defects. In addition, we found that the expression levels of pro-apoptotic genes were elevated in the hearts of Jazf1-Tg mice. These findings suggest that Jazf1 overexpression may induce heart failure symptoms through the upregulation of pro-apoptotic genes in cardiomyocytes.

A new strain of Arthrinium phaeospermum isolated from Carex kobomugi Ohwi is capable of gibberellin production.

Plant growth-promoting endophytic fungi with gibberellin-producing ability were isolated from the roots of Carex kobomugi Ohwi, a common sand-dune plant, and bioassayed for plant growth-promotion. A new strain, Arthrinium phaeospermum KACC43901, promoted growth of waito-c rice and Atriplex gemelinii. Analysis of its culture filtrate showed the presence of bioactive GA(1) (0.5 ng/ml), GA(3) (8.8 ng/ml), GA(4) (4.7 ng/ml) and GA(7) (2.2 ng/ml) along with physiologically inactive GA(5) (0.4 ng/ml), GA(9) (0.6 ng/ml), GA(12) (0.4 ng/ml), GA(15) (0.4 ng/ml), GA(19) (0.9 ng/ml) and GA(24) (1.8 ng/ml). The fungal isolate was identified through sequence homology and phylogenetic analysis of 18S rDNA (internal transcribed region).

Plant growth promotion and Penicillium citrinum.

BACKGROUND: Endophytic fungi are known plant symbionts. They produce a variety of beneficial metabolites for plant growth and survival, as well as defend their hosts from attack of certain pathogens. Coastal dunes are nutrient deficient and offer harsh, saline environment for the existing flora and fauna. Endophytic fungi may play an important role in plant survival by enhancing nutrient uptake and producing growth-promoting metabolites such as gibberellins and auxins. We screened roots of Ixeris repenes (L.) A. Gray, a common dune plant, for the isolation of gibberellin secreting endophytic fungi. RESULTS: We isolated 15 endophytic fungi from the roots of Ixeris repenes and screened them for growth promoting secondary metabolites. The fungal isolate IR-3-3 gave maximum plant growth when applied to waito-c rice and Atriplex gemelinii seedlings. Analysis of the culture filtrate of IR-3-3 showed the presence of physiologically active gibberellins, GA1, GA3, GA4 and GA7 (1.95 ng/ml, 3.83 ng/ml, 6.03 ng/ml and 2.35 ng/ml, respectively) along with other physiologically inactive GA5, GA9, GA12, GA15, GA19, GA20 and, GA24. The plant growth promotion and gibberellin producing capacity of IR-3-3 was much higher than the wild type Gibberella fujikuroi, which was taken as control during present study. GA5, a precursor of bioactive GA3 was reported for the first time in fungi. The fungal isolate IR-3-3 was identified as a new strain of Penicillium citrinum (named as P. citrinum KACC43900) through phylogenetic analysis of 18S rDNA sequence. CONCLUSION: Isolation of new strain of Penicillium citrinum from the sand dune flora is interesting as information on the presence of Pencillium species in coastal sand dunes is limited. The plant growth promoting ability of this fungal strain may help in conservation and revegetation of the rapidly eroding sand dune flora. Penicillium citrinum is already known for producing mycotoxin citrinin and cellulose digesting enzymes like cellulase and endoglucanase, as well as xylulase. Gibberellins producing ability of this fungus and the discovery about the presence of GA5 will open new aspects of research and investigations.