molecular techniques
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2023 ◽  
Vol 83 ◽  
B. R. R. M. Nassau ◽  
P. S. C. Mascarenhas ◽  
A. G. Guimarães ◽  
F. M. Feitosa ◽  
H. M. Ferreira ◽  

Abstract The inheritance of the seedless fruit characteristic of Annona squamosa has not yet been explained. Molecular techniques may aid breeding programs, mainly in the assisted selection of the target gene. The INO gene may be related to seed development in these fruits. The objective of the present paper was to investigate the inheritance of seedlessness in the 'Brazilian seedless' sugar apple and INO gene conservation in Annona squamosa and Annona cherimola x Annona squamosa genotypes by assessing their homology with the INO database genes. The F1 generation was obtained by crossing the mutant 'Brazilian seedless' (male genitor) (P1) with the wild-type A. squamosa with seeds (M1 and M2, female genitors). The INO gene was studied in mutant and wild-type A. squamosa (P1, M1, M2 and M3) and in the Gefner atemoya (A. cherimola x A. squamosa) (M4) cultivar. The DNA was extracted from young leaves, and four sets of specific primers flanking the INO gene were amplified. The seedless characteristic was identified as stenospermatic in the fruits of parental P1, suggesting monogenic inheritance with complete dominance. High sequence similarity of the INO gene amplifications in the sugar apple accessions (M1, M2, M3) and the atemoya cultivar Gefner (M4) reinforces the hypothesis of their conservation.

2023 ◽  
Vol 83 ◽  
L. A. Can-Herrera ◽  
C. D. Gutierrez-Canul ◽  
M. A. A. Dzul-Cervantes ◽  
O. F. Pacheco-Salazar ◽  
J. D. Chi-Cortez ◽  

Abstract Isla Arena is located in the coordinate 20° 70´ N - 90° 45´ W, from Campeche, Mexico. In these estuaries, the ocean mixes with fresh water, and ecosystems are concentrated where petenes and pink flamingos proliferate. Crustaceans and mollusks abound in the sea. Despite its enormous marine wealth, there are no studies carried out on which halophilic microorganisms are present in these waters. In this work, the diversity and structure of the microbial community was investigated through a metagenomics approach and corroborated for sequencing of 16S rRNA genes. It was found that the phylum Fimicutes predominates with more than 50%, in almost the same proportion of the class Bacilli and with almost 41% of relative abundance of the order Bacillales. The sequencing results showed that one of the samples presented a high percentage of similarity (99.75%) using the Nucleotide BLAST program with a peculiar microorganism: Bacillus subtilis. This microorganism is one of the best characterized bacteria among the gram-positive ones. Our results demonstrate that B. subtilis can be an efficient source of proteases, lipases and cellulases, from halophilic microbial communities located in poorly explored areas.

Pathogens ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 106
Marcos Rogério André ◽  
Ana Cláudia Calchi ◽  
Maria Eduarda Chiaradia Furquim ◽  
Isabela de Andrade ◽  
Paulo Vitor Cadina Arantes ◽  

Even though the epidemiology of tick-borne agents (TBA) in dogs has been extensively investigated around the world, the occurrence, vectors involved, and molecular identity of these agents in cats remains elusive in many regions. Among TBA, Ehrlichia, Anaplasma, Babesia, Cytauxzoon, and Hepatozoon are responsible for diseases with non-specific clinical signs in cats, making essential the use of molecular techniques for accurate diagnosis and proper treatment. The present work aimed to investigate the occurrence and molecular identity of tick-borne agents (Ehrlichia, Anaplasma, Babesia/Theileria, Cytauxzoon, and Hepatozoon) in cats from southeastern (states of São Paulo (SP) and Minas Gerais (MG)) and northern (state of Rondônia (RO)) Brazil. For this purpose, 390 blood samples were collected from domiciled cats in MG (n = 155), SP (n = 151), and RO(n = 84) states, submitted to DNA extraction and PCR assays for Ehrlichia spp. (dsb gene), Anaplasma spp. (rrs gene), piroplasmids (18S rRNA gene), and Hepatozoon spp. (18S rRNA gene), sequencing, and phylogenetic inferences. The overall positivity for Anaplasma spp., Ehrlichia spp., Babesia/Theileria spp., Cytauxzoon spp., and Hepatozoon spp. were 7.4% (12.3% (MG) and 6.6% (SP)), 2% (4.5% (MG) and 0.6% (SP)), 0.7% (0.6% (MG), 0.6% (SP) and 1.2% (RO)), 27.2% (41.9% (MG), 24.5% (SP) and 4.8% (RO), and 0%, respectively. The phylogenetic analysis grouped the obtained sequences with ‘Candidatus Anaplasma amazonensis’, A. platys, B. vogeli, and Cytauxzoon sp. previously detected in wild felids from Brazil. qPCR specific for E. canis based on the dsb gene confirmed the molecular identity of the detected ehrlichial agent. The present study expanded the list and geographical distribution of hemoparasites in cats. ‘Candidatus Anaplasma amazonensis’, recently detected in sloths from northern Brazil, was described for the first time in cats. This is the first report of piroplasmids infecting cats in northern Brazil. Coinfection by Cytauxzoon and other TBA (Ehrlichia, Anaplasma, and B. vogeli) reported in the present study raises the need for veterinary practitioners’ awareness of cats parasitized by multiple TBA.

2022 ◽  
Vol 2 ◽  
Sefinew Tilahun ◽  
Marye Alemu ◽  
Mesfin Tsegaw ◽  
Nega Berhane

Ginger diseases caused by fungal pathogens have become one of the most serious problems causing reduced production around the world. It has also caused a major problem among farmers in different parts of Ethiopia resulting in a huge decline in rhizome yield. However, the exact causative agents of this disease have not been identified in the state. Although there are few studies related to pathogenic fungus identification, molecular level identification of fungal pathogen was not done in the area. Therefore, this study was undertaken to isolate and characterized the fungal causative agent of ginger disease from the diseased plant and the soil samples collected around the diseased plant from Chilga district, Gondar, Ethiopia. Samples from infected ginger plants and the soil around the infected plant were collected. Culturing and purification of isolates were made using Potato Dextrose Agar supplemented with antibacterial agent chloramphenicol. The morphological characterization was done by structural identification of the isolates under the microscope using lactophenol cotton blue stains. Isolated fungi were cultured and molecular identification was done using an internal transcribed spacer (ITS) of ribosomal DNA (rDNA). A total of 15 fungal morphotypes including 11 Aspergillus spp. (73.3%), 2 Penicillium spp. (13.3%), and single uncultured fungus clone S23 were isolated from the samples representing all the plant organs and the soil. Aspergillus spp. (73.3%) was the most common and seems to be the major causative agent. To the best of our knowledge, this is the first report of ginger pathogenic fungi in Ethiopia identified using ITS rDNA molecular techniques. This study will lay foundation for the development of management strategies for fungal diseases infecting ginger.

Antibiotics ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 103
Minhian Chai ◽  
Muhammad Zikree Sukiman ◽  
Amirah Huda Kamarun Baharin ◽  
Insyirah Ramlan ◽  
Lennard Zhunhoong Lai ◽  

Staphylococcus aureus (S. aureus) infections, particularly methicillin-resistant Staphylococcus aureus (MRSA) in humans and animals, have become a significant concern globally. The present study aimed to determine the prevalence and antibiogram of S. aureus isolated from animal handlers in Peninsular Malaysia. Furthermore, the genotypic characteristics of S. aureus isolates were also investigated. Nasal and oral swab samples were collected from 423 animal handlers in Peninsular Malaysia. The antibiogram profiles of S. aureus against 18 antibiotics were established using a Kirby–Bauer test. The genotypic profile of S. aureus, including the presence of antimicrobial resistance (AMR), virulence genes and spa genotypes, was investigated using molecular techniques. The overall carriage rate of S. aureus, MRSA and MDRSA was 30.5%, 1.2% and 19.4%, respectively. S. aureus was highly resistant against penicillin (72.3%) and amoxicillin (52.3%). Meanwhile, gentamicin and linezolid were fully effective against all the isolated S. aureus from animal handlers. It was observed that animal handlers with close exposure to poultry were more likely to carry S. aureus that is resistant to tetracycline and erythromycin. S. aureus isolates harboured tetracycline resistance (tetK, tetL and tetM), erythromycin resistance (ermA, ermB, ermC and msrA) and immune evasion cluster (IEC) genes (scn, chp, sak, sea and sep). Seventeen different spa types were detected among the 30 isolates of MDRSA, with t189 (16.7%) and t4171 (16.7%) being the predominant spa type, suggesting wide genetic diversity of the MDRSA isolates. The present study demonstrated the prevalence of S. aureus strains, including MRSA and MDRSA with various antimicrobial resistance and genetic profiles from animal handlers in Peninsular Malaysia.

2022 ◽  
Vol 10 (1) ◽  
pp. 165
Violah Jepkogei Kemboi ◽  
Carolyne Kipkoech ◽  
Moses Njire ◽  
Samuel Were ◽  
Mevin Kiprotich Lagat ◽  

Globally, Ralstonia solanacearum (Smith) is ranked one of the most destructive bacterial pathogens inducing rapid and fatal wilting symptoms on tomatoes. Yield losses on tomatoes vary from 0 to 91% and most control measures are unaffordable to resource-poor farmers. This study investigated the antimicrobial activities of chitin and chitosan extracted from black soldier fly (BSF) pupal exuviae against R. solanacearum. Morphological, biochemical, and molecular techniques were used to isolate and characterize R. solanacearum for in vitro pathogenicity test using disc diffusion technique. Our results revealed that BSF chitosan significantly inhibited the growth of R. solanacearum when compared to treatments without chitosan. However, there was no significant difference in the antibacterial activities between BSF and commercial chitosan against R. solanacearum. Soil amended with BSF-chitin and chitosan demonstrated a reduction in bacterial wilt disease incidence by 30.31% and 34.95%, respectively. Whereas, disease severity was reduced by 22.57% and 23.66%, when inoculated tomato plants were subjected to soil amended with BSF chitin and chitosan, respectively. These findings have demonstrated that BSF pupal shells are an attractive renewable raw material for the recovery of valuable products (chitin and chitosan) with promising ability as a new type of eco-friendly control measure against bacterial wilt caused by R. solanacearum. Further studies should explore integrated pest management options that integrate multiple components including insect-based chitin and chitosan to manage bacterial wilt diseases, contributing significantly to increased tomato production worldwide.

2022 ◽  
Vol 5 (4) ◽  
pp. e202101203
Yasunori Park ◽  
Rachael A West ◽  
Pranujan Pathmendra ◽  
Bertrand Favier ◽  
Thomas Stoeger ◽  

Nucleotide sequence reagents underpin molecular techniques that have been applied across hundreds of thousands of publications. We have previously reported wrongly identified nucleotide sequence reagents in human research publications and described a semi-automated screening tool Seek & Blastn to fact-check their claimed status. We applied Seek & Blastn to screen >11,700 publications across five literature corpora, including all original publications in Gene from 2007 to 2018 and all original open-access publications in Oncology Reports from 2014 to 2018. After manually checking Seek & Blastn outputs for >3,400 human research articles, we identified 712 articles across 78 journals that described at least one wrongly identified nucleotide sequence. Verifying the claimed identities of >13,700 sequences highlighted 1,535 wrongly identified sequences, most of which were claimed targeting reagents for the analysis of 365 human protein-coding genes and 120 non-coding RNAs. The 712 problematic articles have received >17,000 citations, including citations by human clinical trials. Given our estimate that approximately one-quarter of problematic articles may misinform the future development of human therapies, urgent measures are required to address unreliable gene research articles.

Neha Thakur ◽  
S.N. Vasudevan ◽  
B.V. Tembhurne ◽  
S.R. Doddagoudar

Background: Molecular markers are the landmarks on DNA that identifies a particular sequence of base pairs coding for a character. SCAR (Sequenced Characterized Amplified Region) markers are proving to be more effective in identification of genotypes as they are PCR based co-dominant markers. In view of plant variety registration under PPV and FRA, 2001 molecular characterization/ identification of the variety is essential to ascertain the trueness of the variety, hence present studies have been planned and executed. Methods: In present investigation CGMS based chilli hybrid UARChH42 and its parental lines were identified using molecular techniques. A line, B line, R line and hybrid seedlings were used for DNA extraction and characterized on the basis of polymorphism with respect to sterility or fertility by using SCAR markers. Result: P1 and P2 and coxII-SCAR which are sterility specific markers could amplify A line and hybrid showing a definite band. CRF-SCAR 870 is a fertility specific marker amplified R line and hybrid. Also CMS-SCAR 130 and CMS-SCAR 130/140 were able to identified A line, B line, R line and hybrid. Two of the markers viz. orf-456-SCAR atp6-SCAR and were not able to specify any case of parental lines or hybrid identification. Hybridity of UARChH42 (JCH42) chilli hybrid was determined by any similarity in the banding pattern with any of its parent. This study would help in the fulfillment of the requirements of protection of plant varieties and farmers’ rights authority (PPVFRA), New Delhi for registration purpose.

2022 ◽  
Vol 9 (1) ◽  
pp. 21
Vena Chupia ◽  
Jirapat Ninsuwon ◽  
Kakanang Piyarungsri ◽  
Chollada Sodarat ◽  
Worapat Prachasilchai ◽  

Dermatophytosis is a disease caused by dermatophytes, a group of fungi that can cause disease both in humans and animals. The important genera that are pathogenic in animals include Trichophyton and Microsporum. Microsporum canis is an important species because it can cause zoonosis and is commonly found in domestic animals. Cats, which live very close to humans, may expose humans to this pathogen. This research focused on the epidemiology of M. canis found in cats. Hair samples were collected via the Mackenzie technique from cats with and without skin lesions, preliminarily examined with 10% KOH preparation, and cultured for fungal identification. Samples were confirmed with molecular techniques including polymerase chain reaction, gel electrophoresis, and sequencing. Samples were collected from 138 cats located in 93 households, 43 from cats with skin lesions (31.16%) and 95 from cats without skin lesions (68.84%). Eighteen cats with lesions (13.04%) and ten cats without lesions (7.2%) were found to carry M. canis. In eleven of the eighteen cats both with skin lesions and positive for M. canis (61.11%), the pathogen was found both at the site of the lesion and at other sites in the body. Because the pathogen can be found in the hair of cats with and without skin lesions, owners, keepers, veterinarians, and others who come into contact with these animals are at risk of infection if they are not aware or do not take precautions after contact with them.

Earth ◽  
2022 ◽  
Vol 3 (1) ◽  
pp. 45-71
Dhurba Neupane ◽  
Pramila Adhikari ◽  
Dwarika Bhattarai ◽  
Birendra Rana ◽  
Zeeshan Ahmed ◽  

Climate prediction models suggest that agricultural productivity will be significantly affected in the future. The expected rise in average global temperature due to the higher release of greenhouse gases (GHGs) into the atmosphere and increased depletion of water resources with enhanced climate variability will be a serious threat to world food security. Moreover, there is an increase in the frequency and severity of long-lasting drought events over 1/3rd of the global landmass and five times increase in water demand deficits during the 21st century. The top three cereals, wheat (Triticum aestivum), maize (Zea mays), and rice (Oryza sativa), are the major and staple food crops of most people across the world. To meet the food demand of the ever-increasing population, which is expected to increase by over 9 billion by 2050, there is a dire need to increase cereal production by approximately 70%. However, we have observed a dramatic decrease in area of fertile and arable land to grow these crops. This trend is likely to increase in the future. Therefore, this review article provides an extensive review on recent and future projected area and production, the growth requirements and greenhouse gas emissions and global warming potential of the top three cereal crops, the effects of climate change on their yields, and the morphological, physiological, biochemical, and hormonal responses of plants to drought. We also discuss the potential strategies to tackle the effects of climate change and increase yields. These strategies include integrated conventional and modern molecular techniques and genomic approach, the implementation of agronomic best management (ABM) practices, and growing climate resilient cereal crops, such as millets. Millets are less resource-intensive crops and release a lower amount of greenhouse gases compared to other cereals. Therefore, millets can be the potential next-generation crops for research to explore the climate-resilient traits and use the information for the improvement of major cereals.

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