scholarly journals Morphological and Molecular Identification of Mycobiota Associated with Juniperus monosperma (Engelm.) Sarg. Blight in Mexico

2020 ◽  
pp. 1-7
Author(s):  
Ma. E. Galindo-Cepeda ◽  
A. Sánchez-Arizpe ◽  
E. Castro-del Ángel ◽  
S. F. Velázquez-Morales ◽  
J. L. Arispe-Vázquez
Keyword(s):  
Dna Extraction ◽  
Molecular Identification ◽  
Culture Medium ◽  
Dna Amplification ◽  
Agarose Gel ◽  
Juniperus Monosperma ◽  
Morphological Criteria ◽  
Search Tool ◽  
Alternaria Sp ◽  
Aspergillus Sp

The aim of the investigation was to identify the agents associated with blight in Juniperus monosperma at Lirios region, Arteaga, Coahuila. Botanical material was collected at Lirios, Arteaga, and taken to the laboratory. Pathogens were isolated in ADP culture medium and identifying by morphological criteria and molecular using primers ITS1 and ITS4. DNA extraction by the Dellaporta method, the visualization of obtained DNA was performed by electrophoresis on a 2% (p/v) agarose gel. DNA quantification was performed on a NanoDrop 1000 spectrophotometer, and DNA amplification was carried out in the Veriti thermocycler. Obtained sequences were aligned and compared with those available in the GenBank database of the National Center for Biotechnology Information (NCBI), using the BLAST algorithm (Basic Local Aligment Search Tool) to find conserved sequences. Pathogens were inoculated in stems of J. monosperma. The agents associated with blight were Alternaria sp., Aspergillus sp. and Rosellinia sp. and the sequence obtained compared with BLAST coincided only with Alternaria sp. and Aspergillus sp. with the access codes KP027305.1 and MG551283.1, respectively. According to data taken in field, Alternaria sp. behaved as the highest prevalence species and severity associated with blight in J. monosperma, and Rosellinia sp. in laboratory conditions.

scholarly journals Evaluation and Application of an Efficient Plant DNA Extraction Protocol in Laboratory and Field Testing

2020 ◽  
Author(s):  
Qi Wang ◽  
Xiaoxia Shen ◽  
Tian Qiu ◽  
Wei Wu ◽  
Zhian Wang ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Molecular Identification ◽  
Filter Paper ◽  
Biological Samples ◽  
Gc Content ◽  
Dna Amplification ◽  
Nucleic Acid Amplification ◽  
Lamp Method ◽  
Paper Strip ◽  
Cellulose Filter

Abstract Background CTAB has been considered as the standard protocol for DNA extraction. But the complex and time-consuming procedures can’t meet the needs of rapid molecular identification. The method of using cellulose filter paper strips to transfer the DNA in the plant tissue lysate to the nucleic acid amplification system shortening the DNA extraction time to 30 s. However, cellulose filter paper strips have some shortcomings that cannot be put into widespread use. And the data supporting the rapidly purification of DNA by cellulose filter paper is not sufficient.ResultsIn the study, the published filter paper strip was modified by sticking the filter paper on the PVC sheet. This modified method is named EZ-D, for easy DNA extraction. Compared with the original method, the DNA extracted by EZ-D is more efficient in PCR amplification. We also came up with a new DNA extraction buffer, which exhibited higher DNA extraction efficiency. When compared with classic CTAB, EZ-D also showed great advantages for higher efficiency, easier protocol and lower cost. PCR analyses showed that DNA extracted from several types of plants by EZ-D were appropriate for specific identification of biological samples. PCR using DNA extracted by EZ-D was sensitive enough to detect 0.1 ng/μL. Evaluation of the EZ-D showed that the DNA extracts can be successfully amplified by PCR reaction for the DNA fragments up to 3000 bp in length and up to 80% in GC content. EZ-D was successfully used for DNA extraction from a variety of plant species and plant tissues. Moreover, when EZ-D was combined with the loop-mediated isothermal amplification (LAMP) method, DNA identification of biological samples could be achieved in an equipment-free way. Conclusion Combined with DNA amplification technology, EZ-D protocol is a rapid, specific and sensitive method for molecular identification of plant samples. In addition, EZ-D method is the first application of cellulose filter paper in the identification of genetically modified crops and traditional Chinese medicine ultra-fine powder. In terms of practicability, EZ-D has realized the popularization of cellulose filter paper for rapid DNA purification in laboratories and markets.

scholarly journals Presence and importance of saprophyte fungal organisms on dog skin

2007 ◽  
pp. 261-265 ◽  
Author(s):  
Igor Stojanov ◽  
Sandra Jaksic ◽  
Jasna Prodanov
Keyword(s):  
Considerable Increase ◽  
Clinical Symptoms ◽  
Systemic Mycosis ◽  
Skin Changes ◽  
Contaminated Food ◽  
Fungal Organisms ◽  
Alternaria Sp ◽  
Fusarium Sp ◽  
Aspergillus Sp ◽  
One Year

Dogs are animals that are most often kept as pets in the cities. Their health problem may be the cause of infections of humans and animals. Skin changes and etiology factors present important segment of the diseases that disturb health of the pets. The objective of this work was mycology examination of scarifications and skin swabs from dogs with clinical symptoms. The aim was to find out which fungi species can be isolated from the changed parts of the skin, and whether is possible that, besides dermatophyte, saprophyte fungi from the environment may also be the cause of the changes, and to reveal their effect on the host. During a one year period, 67 swabs and scarifications from dogs were examined to detect the presence of fungi. The samples were streaked on Sabourdaud's dextrose agar and incubated for 10-21 days at 25?C. In microscopis examination according to their shape, and color, the colonies were identified as conidia, macroconidia and conidiaophora. From 59, of total 67 samples, the following saprophyte fungi were isolated: Aspergillus sp., Penicillium sp., Alternaria sp., Mucor sp. and Fusarium sp. Occurrence of these fungi means that a considerable increase of this microbiological flora may be expected in homes of the owners. This may be the cause of systemic mycosis and allergies in animals and humans, as well as a possibility of contaminated food and incidence of mycotoxicosis.

scholarly journals Phytosanitary Quality of Genotypes of Wheat Seeds Used Northern Paraná State

2020 ◽  
Vol 12 (6) ◽  
pp. 57
Author(s):  
Jacqueline Dalbelo Puia ◽  
Leandro Camargo Borsato ◽  
Marilize Cristina Gonçalves de Oliveira ◽  
Adriano Thibes Hoshino ◽  
Marcelo Giovanetti Canteri ◽  
...  
Keyword(s):  
Quality Evaluation ◽  
Morphological Characteristics ◽  
Storage Fungi ◽  
Health Quality ◽  
Alternaria Sp ◽  
Low Incidence ◽  
Aspergillus Sp ◽  
Rhizopus Sp ◽  
Wheat Seeds

Wheat seeds can be infested and/or infected by microorganisms that might cause deterioration of this propagation structure. The aim of this study was to evaluate the health quality of sixteen wheat genotypes grown in northern Paraná. Therefore, seeds of each genotype were submitted to the blotter test with 16 repetitions, 400 seeds per sample, for phytosanitary quality evaluation. The identification of the fungi was performed based on their morphological characteristics and quantified data. The results revealed variations in incidence, with 20 fungi genera in the analyzed samples. The fungi Rhizopus sp., Aspergillus sp., Penicillium sp. and Bipolaris sp. were found in 100% of the analyzed samples, while Mucor sp. and Alternaria sp. were in 89% and 78% of the samples, respectively. The main pathogens that cause diseases in the aerial part of wheat were not found, or were low incidence in all materials analyzed. The pathogens with the highest incidence associated with wheat seeds were groups of storage fungi and known to produce mycotoxins.

scholarly journals In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bactéria

2019 ◽  
Vol 49 ◽  
Author(s):  
Ana Paula de Azevedo Pasqualini ◽  
Gabriela Xavier Schneider ◽  
Hugo Pacheco de Freitas Fraga ◽  
Luiz Antonio Biasi ◽  
Marguerite Quoirin
Keyword(s):  
Molecular Identification ◽  
Culture Medium ◽  
Bacterial Isolate ◽  
Fungal Growth ◽  
Endophytic Bacterium ◽  
Liquid Culture Medium ◽  
Rdna Sequencing ◽  
In Vitro Establishment ◽  
Bambusa Oldhamii

ABSTRACT In plant micropropagation, the establishment stage is difficult, due to the presence of microorganisms in tissues from field-grown matrices, especially for bamboo. This study aimed to establish an efficient asepsis protocol for Bambusa oldhamii explants from field plants, as well as to carry out the molecular identification of a possible endophytic bacterial isolate. The explants were exposed to 70 % alcohol, 1 % sodium hypochlorite (NaOCl), 0.1 % mercuric chloride (HgCl2), thiophanate-methyl (Cercobin®) and chlorhexidine digluconate (2 % Riohex®) in different combinations, and introduced into Murashige and Skoog culture medium (solid or liquid), supplemented or not with 4 mL L-1 of Plant Preservative Mixture (PPMTM), totaling seven treatments. The asepsis and immersion of the explants in the liquid culture medium containing 4 mL L-1 of PPMTM visually inhibited the bacterial and fungal growth, allowed the development of shoots with a mean length of 2.2 cm and posterior subcultures, being the best treatment used for the in vitro establishment of B. oldhamii. The molecular identification of an endophytic bacterium performed by 16S rDNA sequencing allowed to identify the bacterial isolate as Ralstonia sp., with 100 % of similarity, and the phylogenetic analysis grouped it with Ralstonia pickettii. In addition, the bacterial isolate showed to be sensitive to 4 mL L-1 of PPMTM by the minimum inhibitory concentration test.

In Vitro Evaluation of Antifungal Activity of Three Traditionally Used Medicinal Plants; Umbilicus Intermedius Boiss, Cuminum Cyminum and Zingiber Officinale Extracts

2019 ◽  
Author(s):  
Ameneh Takesh ◽  
Mahnaz Fatahinia ◽  
Ali Zarei Mahmoudabadi
Keyword(s):  
Antifungal Activity ◽  
Culture Medium ◽  
Zingiber Officinale ◽  
Diffusion Method ◽  
Disc Diffusion ◽  
Disc Diffusion Method ◽  
Cuminum Cyminum ◽  
Medicinal Plant Extracts ◽  
Alternaria Sp ◽  
Medium Method

Background and Aims: The study aimed to determine the effectiveness of three medicinal plant extracts on fungi with three methods and to compare methods.  Material and methods: This study examined the antifungal properties of cumin (Cuminum cyminum L), ginger (Zingiber officinale Roscoe) and Nafe Venus (Umbilicus intermedius boiss) extracts against fungi including, Aspergillus spp., Penicillium spp., Mucor spp., Stemphylium spp., Drechslera spp., Alternaria spp., Cladosporium spp., and Aureobasidium pullulans. Furthermore, 17 candida isolates including, C. albicans, C. glabrata and C. dubliniensis were tested. In the present study two methods of disc diffusion method, agar wells diffusion method were used for assay. Then, the mixing with culture medium method was used for assessment of the antifungal activity of extracts against Alternaria sp.(as black mold), A. terreus (as hyaline mold) and C. albicans (as yeast) to compare methods as well. Results: No fungi were susceptible to extracts in disc diffusion method and agar wells diffusion method. But, this study showed that in mixing with culture medium method, cumin extract has valuable anti-fungal property and Umbilicus intermedius boiss has the inhibitory properties against the black fungi. Furthermore, it is found that mixing with culture medium method is more efficient than disc and agar well diffusion methods. Alternaria sp. and C. albicans were susceptible and resistant to all extracts.  Conclusions: it is found that mixing with culture medium method is more efficient than disc and agar well diffusion methods and inhibitory potency of the extracts varies according to the type of extraction and their concentration.

scholarly journals Trigona sp. COMO VISITANTE FLORAL E VETOR DE ESPOROS FÚNGICOS PARA GOIABEIRA (Psidium guajava L. – MYRTACEAE)

2004 ◽  
Vol 26 (55) ◽  
Author(s):  
Rodrigo Makowiecky Stuart ◽  
Carolina Lamas ◽  
Ida Chapaval Pimentel
Keyword(s):  
Psidium Guajava ◽  
Penicillium Sp ◽  
Alternaria Sp ◽  
Fusarium Sp ◽  
Aspergillus Sp ◽  
Rhizopus Sp ◽  
Mycelia Sterilia

Este trabalho foi realizado para avaliar a ação de Trigona sp. como visitante floral e vetor de esporos fúngicos para goiabeira (Psidium guajava L.). As observações foram feitas entre dezembro de 2003 e fevereiro de 2004 em flores de goiabeiras do Setor de Ciências Biológicas, Universidade Federal do Paraná. Foi verificado que três gêneros de abelhas estavam visitando as flores de goiaba: Apis melifera, Bombus sp. e Trigona sp.. Entretanto, Trigona demonstrou ser mais freqüente que as outras durante as observações. A freqüência de visitas de Trigona foi maior durante o início da manhã, decrescendo ao longo do dia. A avaliação dos fungos associados a Trigona demonstrou a presença de 11 gêneros distintos: Acremonium sp., Altenaria sp., Aspergillus sp., Colletotrichum sp., Curvularia sp., Fonsecaea sp., Fusarium sp., Mycelia sterilia, Penicillium sp., Phoma sp. e Rhizopus sp.. 48 % destes representam fitopatógenos potenciais como Alternaria sp., Colletotrichum sp., Curvularia sp., Fusarium sp. e Phoma sp.. Estes dados demonstram que o gênero Trigona pode atuar na disseminação de doenças para diversas culturas, funcionando como vetor de esporos fúngicos para outras plantas.

scholarly journals A Protocol for Rapid and Inexpensive Genotyping of Mutant Mice from Dried Blood Spots: An Update

2021 ◽  
Author(s):  
Antonella Romano ◽  
Candida Zuchegna ◽  
Giuseppa Zannini ◽  
Roberta Grillo ◽  
Samantha Messina ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Genomic Dna ◽  
Fragment Length Polymorphism ◽  
Dna Amplification ◽  
Dried Blood Spots ◽  
Nucleotide Polymorphisms ◽  
Single Nucleotide ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Sensitivity Specificity

Abstract Background: Dried blood spot (DBS) testing is a well-known method of bio-sampling by which blood samples are blotted and dried on filter paper. The dried samples can then be analyzed by several techniques such as DNA amplification and HPLC. We have developed a homemade DBS method followed by an alternative protocol for genomic DNA extraction from a drop of blood adsorbed on paper support. This protocol consists of two separate steps: (1) organic DNA extraction from the DBS, followed by (2) DNA amplification by polymerase chain reaction (PCR). The PCR-restriction fragment length polymorphism (PCR-RFLP) is an advantageous and simple approach to detect single nucleotide polymorphisms (SNPs). Results: We have evaluated the efficiency of our method for the extraction of genomic DNA from DBS by testing its performance in genotyping mouse models of obesity and herein discuss the sensitivity, specificity and feasibility of this novel procedure. Conclusions: Our protocol is easy to perform, fast and inexpensive and allows the isolation of pure DNA from a miniscule amount of sample.

scholarly journals Optimization of Various ITS rDNA Amplification Protocol of Yeast Isolated from Giant Honey Beehives (Apis dorsata)

2021 ◽  
Vol 3 (2) ◽  
pp. 80-87
Author(s):  
Chumaidatul Choiriyah, S.Si ◽  
Nirmala Fitria Firdhausi ◽  
Esti Tyastirin ◽  
Yuanita Rachmawati ◽  
Moch. Irfan Hadi
Keyword(s):  
Dna Extraction ◽  
Incubation Time ◽  
Dna Amplification ◽  
Extraction Process ◽  
Optimal Time ◽  
Apis Dorsata ◽  
Molecular Approaches ◽  
Amplification Protocol ◽  
Its Gene ◽  
Dna Purity

Indonesia is a country with high variability of microorganisms, including bacteria, yeast, and fungi. Yeast isolates could be isolated from the honeycomb of Apis dorsata. Molecular approaches were used to identify yeast using ribosomal DNA gene sequences, called the ITS gene. The optimum condition for DNA extractions and amplifications are needed for the successfully of molecular identification. Therefore, it is necessary to optimize the DNA extraction and amplification of several protocols to obtain good identification results. This study aimed to compare the effects of DNA extraction with various temperatures and different amplification protocols. LIPI reference DNA extraction protocol with the boiling method and variations in incubation time of 10, 15, and 20 minutes at a temperature of 98° C. Meanwhile, for the amplification of yeast DNA using a variety of different amplification protocols. The results showed the optimal time of incubation was 10 minutes in K1 isolates with DNA purity of 1.896. meanwhile, for isolates K2, K3, and K4 each with a purity of 2.246, 2.335, and 1.748. optimal DNA amplification results were indicated by the presense of DNA bands for each sample K1, K2, K3, and K4, namely 503, 542, 492, and 526 bp. In this study, it can be concluded that the optimal incubation time for the extraction process is 10 minutes. In addition, the optimal amplification protocol was shown in the DNA bands in all sample.

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