Reidentification of cellulolytic enzyme-producing Trichoderma strains W-10 and G-39

2006 ◽  
Vol 52 (6) ◽  
pp. 570-574
Author(s):  
Ching-Fu Lee ◽  
Daniel Yuen Teh Liu ◽  
Ming Tsong Lai ◽  
Tzong-Hsiung Hseu

Strain W-10, originally identified as Trichoderma koningii, and its supposed mutant G-39, published for production and gene expression of cellulase and xylanase, demonstrated morphological characteristics distinct from those of T. koningii, respectively. To clarify the identification derived from morphological characteristics, several methods were used, including electrophoretic karyotyping, internal transcribed spacer (ITS) analysis of rDNA, and polymerase chain reaction (PCR) fingerprinting using the universal primer L45. All the molecular characteristics showed that strains G-39 and W-10 were identical to T. reesei and T. longibrachiatum, respectively. The results strongly supported that T. koningii G-39 and W-10 should be reassigned as T. reesei and T. longibrachiatum, respectively. Strain G-39 should be considered a mutant from T. reesei QM9414 whose spores were contaminated with those of strain W-10 during a laboratory operation. According to this, we declare that T. koningii G-39 and W-10 must be renamed as T. reesei and T. longibrachiatum, respectively.Key words: PCR fingerprinting, electrophoretic karyotypes, ITS, Trichoderma.

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 689
Author(s):  
Dina M. Metwally ◽  
Shurug A. Albasyouni ◽  
Ibrahim A.H. Barakat ◽  
Isra M. Al-Turaiki ◽  
Amal M. Almuhanna ◽  
...  

Heads of sheep (n = 600) and goats (n = 800) slaughtered at Al-Aziziah Abattoir in Riyadh, Saudi Arabia, were inspected for the presence of O. ovis larvae (L). Heads were split along the longitudinal axes, and larvae (L1, L2, and L3) were gathered. The infestation rate was significantly higher in goats (44.5%; 356/800) than that in sheep (22.3%; 134/600). Out of the 151 collected larvae from sheep, 0% were L1, 1.3% were L2, and 98.7% were L3. Out of the total of 468 larvae from goats, 0% were L1, 1.2% were L2, and 98.8% were L3. The infestation rate was significantly higher in males than that in females. Myiasis-causing larvae collected from Riyadh, Saudi Arabia, were authenticated as O. ovis, according to morphological characteristics. Polymerase chain reaction (PCR) amplification of a partial fragment (600 bp) of the mitochondrial cytochrome c oxidase subunit I (mtCOI) gene further confirmed the species. Phylogenetic analysis based on the partial mtCOI gene sequence demonstrated that 23 unique sequences showed high similarity based on nucleotide pairs of O. ovis accessions retrieved from GenBank.


2012 ◽  
Vol 44 (2) ◽  
pp. 7 ◽  
Author(s):  
S. Epis ◽  
M. Montagna ◽  
F. Comandatore ◽  
C. Damiani ◽  
A. Diabaté ◽  
...  

The acetic acid bacterium <em>Asaia</em> spp. was successfully detected in <em>Anopheles arabiensis</em> Patton, 1905, one of the major vector of human malaria in Sub-Saharan Africa. A collection of 45 <em>Asaia</em> isolates in cellfree media was established from 20 individuals collected from the field in Burkina Faso. 16S rRNA universal polymerase chain reaction (PCR) and specific qPCR, for the detection of <em>Asaia</em> spp. were performed in order to reveal the presence of different bacterial taxa associated with this insect. The isolates were typed by internal transcribed spacer-PCR, BOX-PCR, and randomly amplified polymorphic DNA-PCR, proved the presence of different <em>Asaia</em> in <em>A. arabiensis</em>.


Author(s):  
Wan Huang ◽  
Jisheng Zhang ◽  
Lingyi Zeng ◽  
Chengru Yang ◽  
Lining Yin ◽  
...  

BackgroundThis study aimed to determine the molecular characteristics of carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates in a hospital in western Chongqing, southwestern China.MethodsA total of 127 unique CRKP isolates were collected from the Yongchuan Hospital of Chongqing Medical University, identified using a VITEK-2 compact system, and subjected to microbroth dilution to determine the minimal inhibitory concentration. Enterobacteriaceae intergenic repeat consensus polymerase chain reaction and multilocus sequence typing were used to analyze the homology among the isolates. Genetic information, including resistance and virulence genes, was assessed using polymerase chain reaction. The genomic features of the CRKP carrying gene blaKPC-2 were detected using whole-genome sequencing.ResultsST11 was the dominant sequence type in the homology comparison. The resistance rate to ceftazidime-avibactam in children was much higher than that in adults as was the detection rate of the resistance gene blaNDM (p &lt; 0.0001). Virulence genes such as mrkD (97.6%), uge (96.9%), kpn (96.9%), and fim-H (84.3%) had high detection rates. IncF (57.5%) was the major replicon plasmid detected, and sequencing showed that the CRKP063 genome contained two plasmids. The plasmid carrying blaKPC-2, which mediates carbapenem resistance, was located on the 359,625 base pair plasmid IncFII, together with virulence factors, plasmid replication protein (rep B), stabilizing protein (par A), and type IV secretion system (T4SS) proteins that mediate plasmid conjugation transfer.ConclusionOur study aids in understanding the prevalence of CRKP in this hospital and the significant differences between children and adults, thus providing new ideas for clinical empirical use of antibiotics.


2021 ◽  
pp. 3224-3228
Author(s):  
Budi Utomo ◽  
Rimayanti Rimayanti ◽  
Indah Norma Triana ◽  
Amaq Fadholly

Background and Aim: The genetic improvement of cattle through livestock section is based on quantitative, qualitative, and molecular characteristics. This study examined polymorphisms of the melanocortin-4 receptor (MC4R) and leptin genes as a reference for the selection of superior breeds in Madrasin cattle. Materials and Methods: The leptin and MC4R genes of Madrasin cattle were amplified using polymerase chain reaction (PCR); then, restriction fragment length polymorphism of the leptin gene was performed using the restriction enzyme BsaA1, at site 2793 with ACGT point position. Results: The leptin gene was divided into three bands, namely, AA with one fragment (522 bp), CG with two fragments (441 bp and 81 bp), and AG with three fragments (522 bp, 441 bp, and 81 bp). The MCR-4 gene was divided into three bands, namely, 493 bp, 318 bp, and 175 bp. Conclusion: The MC4R and leptin genes can act as molecular markers for growth traits in Madrasin cattle and can be used to genetically optimize and improve growth. The GG allele of the MC4R gene and the AA allele of the leptin gene can be used in Madrasin cattle.


Author(s):  
J.C. De Witte ◽  
I. Deblauwe ◽  
Gill De Deken ◽  
R. De Deken ◽  
M. Madder ◽  
...  

A DNA microarray test based on internal transcribed spacer 1 (ITS1) genotype expression was developed to identify Culicoides species of Northwestern Europe belonging to the Culicoides Obsoletus complex. The assay was designed so as to allow interpretation by the naked eye. False positive and false nega­tive results were eliminated. The need for expensive laboratory equipment and reagents was kept as low as possible, making the technique affordable and feasible for any diagnostic laboratory with polymerase chain reaction (PCR) facilities. The microarray test could be validated through the three ringtests organised by Medreonet. Use of this microarray can improve monitoring adult and immature Culicoides species.


2019 ◽  
Vol 58 (3) ◽  
pp. 401-403 ◽  
Author(s):  
Tomoyuki Iwanaga ◽  
Tsuyoshi Ushigami ◽  
Kazushi Anzawa ◽  
Takashi Mochizuki

Abstract The viability of pathogenic fungi in the scale was investigated during topical administration of 1% luliconazole (LLCZ). Thirteen tinea pedis patients found to be positive on KOH examination were assessed by mycological examinations and quantitative real-time polymerase chain reaction (PCR) targeted internal transcribed spacer (ITS) in ribosomal RNA gene at the initial visit and after 2 and 4 weeks of treatment. Assays showed that the average copy number of ITS DNA had significantly decreased to 22.9% at 2 weeks and 4.8% at 4 weeks compared with the initial visit. LLCZ topical treatment could defeat almost pathogenic dermatophytes in the scales within 4 weeks.


Genome ◽  
1994 ◽  
Vol 37 (4) ◽  
pp. 664-671 ◽  
Author(s):  
Lang Zhuo ◽  
S. L. Sajdak ◽  
R. B. Phillips

Intraspecific variation in the sequence of the transcribed spacer regions of the ribosomal DNA (rDNA) in lake trout was examined by restriction mapping and sequencing of these regions amplified by the polymerase chain reaction. The length of the first internal transcribed spacer region (ITS-1) was 566 bases and the second internal transcribed spacer region (ITS-2) was 368 bases in lake trout. When the 1.4-kb region including the ITS-1, the 5.8S coding region, and the ITS-2 was amplified from 12 individuals from four populations and digested with eight different enzymes only one intraindividual polymorphism was found that occurred in each population. When the amplified ITS-1 region was sequenced from an additional 10 individuals from five populations, no interindividual variation was found in the sequence. A 6-kb portion of the rDNA repeat unit including 1.6 kb of the 18S coding region, the 5′ external spacer region (5′ ETS), and part of the adjacent intergenic spacer was cloned and a restriction map was prepared for these regions in lake trout. No intraspecific variation was found in the region adjacent to the 18S rDNA, which includes the 5′ ETS, although intraspecific and intraindividual length variation was found in the intergenic spacer region 3–6 kb from the 18S. Sequencing of a 609-b segment of the 5′ ETS adjacent to the 18S coding region revealed the presence of two 41-b repeats. The 198-b sequence between the repeats had some similarity to the 18S coding region of other fishes. Primers were designed for amplification of 559 b of the 5′ ETS using the polymerase chain reaction. No intraspecific variation in this region in lake trout was found when the DNA amplified from this region in 12 individuals from four populations was digested with eight restriction enzymes.Key words: ribosomal DNA, internal transcribed spacer regions, 5′ external spacer region, transcribed spacer, lake trout.


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