Molecular analysis and anticancer properties of two identified isolates,Fusarium solani and Emericella nidulans isolated from Wady El-Natron soil in Egypt against Caco-2(ATCC) cell line

Objective:To characterize,identify and investigate the anticancer properties of two new soil fungal isolates,Emericella nidulansand Fusarium solani isolated from Wady El-Natron in Egypt against colon cancer Caco-2(ATCCj cell line.Methods:Soil sample was cultured and two strains were chosen for morphological and phenotypical characterization.Partial sequences of the 18s rRNA gene and the internal transcribed spacer region ITS of the two isolates were amplified by PCR.Phylogenetic tree construction and analysis of the resulted multiple sequences from the two fugal isolates were also carried out.In vitro anticancer activity of the two strains was done against colon Caco-2 cancer cell line.Reverse transcription — PCR was carried out to detect level of expression of p53 in Caco-2 cell line.Results:HF.I displayed morphological and genotypic characteristics most similar to that of Fusarium solani while HF.2 was most similar to Emericella nidulans with high similarity of 99%and 97%respectively.The multiple sequence alignment of the two fungal isolates showed that,the maximum identical conserved domains in the 18s rRNA genes were identified with the nucleotide regions of Slst to 399th base pairs,88th to 525th base pairs respectively.While those in the ITS genes were identified with the nucleotide regions of 88th to 463rd and Slst to 274th.The two isolates showed IC<sub>50 value with（6.24±5.21） and（9.84±0.36） μ g/mL) concentrations respectively at 28h.Reverse transcription- PCR indicated that these cells showed high level of expression for p53 mRNA.Conclusions:The morphology and molecular analysis identified HF.1 and HF.2 to be Fusarium solani and Emericella nidulans;new isolates of anticancer producing fungi from Wady El-Natroon city in Egypt.Treatment with the two isolates caused P53 expression in Caco-2 cell line.These two isolates can be used as an anticancer agents.

Characterization of Amphidinium(Amphidiniales,Dinophyceae)species from the China Sea based on morphological,molecular,and pigment data

Amphidinium species are amongst the most abundant benthic dinoflagellates in marine intertidal sandy ecosystems.Some of them are able to produce a variety of bioactive compounds that can have both harmful effects and pharmaceutical potentials.The diversity of Amphidinium in shallow waters along the Chinese coast was investigated by isolating single cells from sand,coral,and macroalgal samples collected from 2012 to 2020.Their morphologies were subjected to examination using light microscopy(LM)and scanning electron microscopy(SEM).A total of 74 Amphidinium strains were morphologically identified,belonging to 11 species:A.carterae,A.gibbosum,A.operculatum,A.massartii,A.cf.massartii,A.fijiensis,A.pseudomassartii,A.steinii,A.thermaeum,A.theodori,A.tomasii,as well as an undefined species.The last seven species have not been previously reported in Chinese waters.Amphidinium carterae subcladesⅠ,Ⅱ,andⅣwere found in the South China Sea,while subcladeⅢwas only found in the Yellow Sea.Threadlike body scales were observed on the surface of subcladesⅢ and Ⅳ,supporting the idea that A.carterae might contain several different species.Large subunit ribosomal RNA(LSU rRNA)sequencesbased phylogeny revealed two groups(GroupsⅠandⅡ)within Amphidinium,which is consistent with the relative position of sulcus(in touch with cingulum or not).In addition,large differences in morphology and molecular phylogeny between A.operculatum(the type species of Amphidinium)and other species,suggest that a subdivision of Amphidinium might be needed.The pigment profiles of all available strains were analyzed by high performance liquid chromatography(HPLC).Eleven pigments,including peridinin,diadinoxanthin,diatoxanthin,pheophorbide(and pheophorbide a),antheraxanthin,β-carotene,and four different chlorophylls were detected.The high pheophorbide/pheophorbide a ratio in Amphidinium implies that it may be a good candidate as a natural source of photosensitizers,a well-known anticancer drug.