Isolation and Identification of Fungal Endophytes of the Cowpea in Khuzestan Province

Introduction: Endophytes are microorganisms that colonize internal tissues of plants without causing obvious symptoms. This study was conducted to isolate and identify endophytic fungi of the cowpea in Khuzestan province. Materials and methods: During 2016, eight healthy samples of the cowpea plants were collected from the important areas under cultivation in the northern Khuzestan province. The small parts of the roots, stems, leaves and pods were deeply surface sterilized for each samples and plated on Potato-Dextrose-Agar. Sixty fungal isolates obtained in this study were purified by single spore method. Based on morphological characteristics, 21 out of 60 isolates were selected for molecular study. The isolates were grown in Potato-Dextrose-Broth and mycelial biomass was recovered by passing through filter paper. DNA extraction was performed using a phenol- and chloroform- based organic method. The parts of the nrRNA gene (ITS and 28S-D1/D2 regions) were amplified using appropriate primer pair and then sequenced. Results: The isolates were analyzedon the basis of morphological characteristics in combination with BlASTn search algorithm and ITS sequence-based phylogeny. Accordingly, the isolates were identified as follows: Alternaria destruens, Alternaria sp., Curvularia mosaddeghii, Curvularia sp., Fusarium chlamydosporum, F. nygamai, F. falciforme, F. proliferatum, Fusarium sp. Macrophomina phaseolina and Penicillium oxalicum. Discussion and conclusion: Alternaria, Fusarium and Curvularia genera were the most abundant fungal endophytes into cowpea plants, growing in warm climate of the Khuzestan Province. To the best of our knowledge, this is the first study to report endophytic growth of A. destruens, Alternaria sp., M. phaseolina, F. chlamydosporum, F. nygamai, F. falciforme, F. proliferatum and P. oxalicum within cowpea plants

Xenoacremonium palmarum M. Amani, M. Mehrabi-Koushki & R. Farokhinejad 2023, sp. nov.

<i>Xenoacremonium palmarum</i> M. Amani, M. Mehrabi-Koushki & R. Farokhinejad, <i>sp</i>. <i>nov</i>. (Fig. 2) <p> <b>MycoBank</b>: <b>MB849595</b></p> <p> <i>Holotype</i>: IRAN, Khuzestan Province, Ahvaz, isolated from rotten root of <i>Phoenix dactylifera</i> (Palmaceae), May 2021, <i>M. Amani</i> (holotype, IRAN 18302F; ex-type cultures, IRAN 4860C = SCUA-Am-A29).</p> <p> <i>Etymology</i>. The species epithet “ <i>palmarum</i> ” reflects the host plant.</p> <p> <b>Asexual morph on PDA:</b> <i>Hyphae</i> hyaline, septate, branched, 1.5–2.5 µm in diam. <i>Sporulation</i> from lateral phialidic pegs not observed. <i>Conidiophores</i> arose laterally from aerial hyphae, hyaline, straight or slightly curved, subcylindrical, commonly unbranched, but rarely one-times dichotomously branched, 0–2(–3)-septate, smooth, (16.5–)24.5–54.5(–60) × (1.2)1.8–2.6 μm, 95% confidence limits = 34.5–39.5 × 2.1–2.2 μm, (x̅ ± SD = 36.5 ± 9.8 × 2.1 ± 0.3 μm, n = 60), terminating in one or two conidiogenous cells. <i>Conidiogenous cells</i> terminal, hyaline, smooth, cylindrical to subulate, tapering towards apex (11.5–)14.5–37.5(–48) × 1.2–2.5 μm, 95% confidence limits = 21.5–25 × 1.8–1.9 μm, (x̅ ± SD = 23 ± 7 × 1.9 ± 0.3 μm, n = 60), apex 0.6–1.2 μm in diam. <i>Conidia</i> formed abundantly in slimy heads at apex of conidiogenous cells, aseptate, ellipsoidal to fusiform or falcate, curved, smooth, pointed at both ends, (2.9–)3.1–9.9 × 1–2.1 µm, 95% confidence limits = 5.1–6 × 1.3–1.5 μm, (x̅ ± SD = 5.5 ± 1.7 × 1.4 ± 0.3 μm, n = 60). <i>Chlamydospores</i> not observed. <b>Sexual morph:</b> not observed.</p> <p>Culture characteristics—The colonies diameter on PDA, CMA, and OA were 43–46, 56–62, and 19–20 mm after 14 days of incubation at 25 ± 1° C, respectively. Colonies on PDA circular with filiform margin, pale vinaceous to rosy vinaceous, floccose with abundant aerial mycelium; reverse pale vinaceous. Colonies on CMA circular with regular margin, white, floccose, radiate, with aerial mycelium; reverse pale vinaceous. Colonies on OA circular with undulate to entire margin, white, cottony with abundant aerial mycelium; reverse white to pale vinaceous.</p> <p> <i> <i>Additional materials examined</i>.</i> IRAN, Khuzestan Province, Karoon, isolated from rotten root of <i>P. dactylifera</i>, Sep. 2021, M. Amani (SCUA-Am-A29-1, SCUA-Ama-A29-2, and SCUA-Ama-A30); Bushehr Province, Ab pakhsh, isolated from rotten root of <i>P. dactylifera</i>, Sep. 2023, M. Amani <b>(</b> SCUA-Ama-A30-1, SCUA-Ama-A30-2).</p> <p> <i>Notes</i>: Phylogenetically, <i>Xenoacremonium palmarum sp. nov.</i>, grouped with <i>X. allantoideum, X. falcatum</i> and <i>X. minutisporum</i> (Fig. 1). Nucleotide comparison of <i>X. palmarum</i> with <i>X. falcatum</i> revealed that these two species share 97% sequence identity in the <i>tub2</i> gene (335 bp) attributed to 9 SNPs and 2 bp insertion/deletion, and 90 % sequence identity in the <i>tef1α</i> gene (420 bp) attributed to 21 SNPs and 22 bp insertion/deletion. In addition, <i>X. palmarum</i> and <i>X. minutisporum</i> shared 93.2 % sequence identity in the <i>tef1α</i> gene (265 bp) attributed to 10 SNPs and 8 bp insertion/ deletion. Both <i>X. allantoideum</i> and <i>X</i>. <i>palmarum</i> differ from <i>X. falcatum</i> and <i>X. minutisporum</i> in not having lateral phialidic pegs on its somatic hyphae, a feature which has not also been reported for <i>X. recifei</i> (Gams, 1971, Lombard <i>et al.</i> 2015, Roeun <i>et al.</i> 2022, Hou <i>et al</i>. 2023). Conidia in <i>X. falcatum</i> are more curved than those in <i>X. palmarum</i> and <i>X. minutisporum</i> (Gams 1971, Lombard <i>et al.</i> 2015). Furthermore, <i>X. palmarum</i> differs from <i>X. allantoideum</i> in having longer conidia (3.1–9.9 vs 3.6–6 μm) and by the lack of mycelial ropes and verticillately branched conidiophores (Hou <i>et al</i>. 2023).</p>Published as part of <i>Amani, Majid, Farokhinejad, Reza & Mehrabi-Koushki, Mehdi, 2023, Xenoacremonium palmarum sp. nov., a novel species associated with Phoenix dactylifera in Iran, pp. 165-174 in Phytotaxa 632 (2)</i> on pages 169-170, DOI: 10.11646/phytotaxa.632.2.6, <a href="http://zenodo.org/record/10438497">http://zenodo.org/record/10438497</a&gt

Expression and purification of TAT-NDRG2 recombinant protein and evaluation of its anti-proliferative effect on LNCaP cell line

N-myc downstream regulated gene2 (NDRG2) belongs to tumor suppressor protein family of NDRG. Anti proliferative and anti-metastasis of NDRG2 overexpression has been demonstrated in a number of tumors. The aim of this study was to fuse the gene of Trans Activator of Transcription (TAT) protein transduction domain with NDRG2 gene and express and purify TAT-NDRG2 fusion protein in order to investigate the effects of TAT-NDRG2 protein on proliferation and apoptosis of LNCaP prostate carcinoma cell line. pET28a-TAT-NDRG2 and pET28a-NDRG2 plasmids were constructed and transformed into E. coli-BL21(DE3). TAT-NDRG2 and NDRG2 proteins were expressed in the bacteria, purified using affinity chromatography and verified using western blotting. The effects of TAT-NDRG2 and NDRG2 protein treatment on LNCaP cells proliferation and apoptosis were evaluated using MIT assay and AnnexinV, 7-AAD flow cytometry assay, respectively. Western blot analysis confirmed the expression and purification of TAT-NDRG2 and NDRG2 proteins. Treatment of LNCaP cells with TAT-NDRG2 protein increased cell death and induced apoptosis significantly (P < 0.05) compared to control and NDRG2 protein-treated cells. These results suggest that TAT-NDRG2 protein can be considered as a therapeutic modality for the treatment of tumors. (c) 2017 Elsevier Inc. All rights reserved

Fungal Systematics and Evolution: FUSE 5

Thirteen new species are formally described: Cortinarius brunneocarpus from Pakistan, C. lilacinoarmillatus from India, Curvularia khuzestanica on Atriplex lentiformis from Iran, Gloeocantharellus neoechinosporus from China, Laboulbenia bernaliana on species of Apenes, Apristus, and Philophuga (Coleoptera, Carabidae) from Nicaragua and Panama, L. oioveliicola on Oiovelia inachadoi (Hemiptera,Veliidae) from Brazil, L. termiticola on Macrotermes subhyalinus (Blattodea, Termitidae) from the DR Congo, Pluteus cutefractus from Slovenia, Rhizoglomus variabile from Peru, Russula phloginea from China, Stagonosporopsis flacciduvarum on Vitis vinifera from Italy, Strobilomyces huangshanensis from China, Uroinyces klotzschianus on Rumex dentatus subsp. klotzschianus from Pakistan.The following new records are reported: Alternaria calendulae on Calendula officinalis from India; A. tenuissima on apple and quince fruits from Iran; Candelariella oleaginescens fromTurkey; Didymella americana and D. calidophila on Vitis vinifera from Italy; Lasiodiplodia theobromae causing tip blight of Dianella tasmanica variegata' from India; Marasmiellus subpruinosus from Madeira, Portugal, new for Macaronesia and Africa; Mycena albidolilacea, M. tenuispinosa, and M. xantholeuca from Russia; Neonectria neomacrospora on Madhuca longifolia from India; Nothophoma quercina on Vitis vinifera from Italy; Plagiosphaera immersa on Urtica dioica from Austria; Rinodina sicula from Turkey; Sphaerosporium lignatile from Wisconsin, USA; and Verrucaria murina from Turkey. Multi-locus analysis of ITS, LSU, rpbl,tefl sequences revealed that P immersa, commonly classified within Gnomoniaceae (Diaporthales) or as Sordariomycetes incertae sedis, belongs to Magnaporthaceae (Magnaporthales). Analysis of a six-locus Ascomycota-wide dataset including SS