scholarly journals Specificity and sensibility of primer pair in the detection of Colletotrichum gossypii var. cephalosporioides in cotton seeds by PCR technique

2020 ◽  
Vol 42 ◽  
Author(s):  
Mirella Figueiró de Almeida ◽  
Sarah da Silva Costa ◽  
Iara Eleutéria Dias ◽  
Carolina da Silva Siqueira ◽  
José da Cruz Machado
Keyword(s):  
Cotton Seed ◽  
Water Restriction ◽  
Seed Certification ◽  
Certification Programs ◽  
Seed Health ◽  
Target Dna ◽  
Cotton Seeds ◽  
Primer Sets ◽  
Specific Fragment ◽  
Cotton Plantations

Abstract: Cotton Ramulosis (Gossypium hirsutum) is an important disease affecting cotton plantations in Brazil, and its causal agent, Colletotrichum gossypiivar.cephalosporioides(Cgc), according to the Brazilian phytosanitary authority, was considered a regulated non quarantine pest. It makes this microorganism subject to standardization in seed certification programs. The current seed health testing for detecting that pathogen in seed samples does not provide reliable results for routine analysis. On this paper, attempts were made to design specific primers for detection of Cgc associated with cotton seed. Two primer sets were selected based on the analysis of a multiple alignment of gene’s sequence encoding the glyceraldehyde 3-phosphate dehydrogenase from Cgc, C. gossypii and reference strains of the C. gloeosporioides species complex. The conserved sites unique to Cgc strains were used to design specific fragment of 140 bp. The primer specificity was confirmed by using other fungi. The primers produced a detectable band of target DNA of Cgc in all inoculum potentials of the pathogen artificially inoculated by the water restriction technique. The developed primer pair represents, therefore, a reliable and rapid mean to diagnose the Ramulosis agent in cotton seed.

scholarly journals Evaluation of inoculum potential of pathogens in seeds: relation to physiological quality and DNA quantification by qPCR

2017 ◽  
Vol 39 (3) ◽  
pp. 224-233 ◽  
Author(s):  
Marina de Resende Faria Guimarães ◽  
Carolina da Silva Siqueira ◽  
José da Cruz Machado ◽  
Sueny Kelly Santos de França ◽  
Gabriel Castanheira Guimarães
Keyword(s):  
Germination Percentage ◽  
Seed Vigor ◽  
Soybean Seeds ◽  
Inoculum Potential ◽  
Water Restriction ◽  
Maize Seeds ◽  
Seed Health ◽  
Physiological Quality ◽  
Cotton Seeds ◽  
Stenocarpella Maydis

Abstract: Given what is already known in regard to seed health and the availability of molecular methods for detection of the pathogens Stenocarpella maydis and Stenocarpella macrospora in maize seeds, Colletotrichum gossypii var. cephalosporioides in cotton seeds, and Corynespora cassiicola in soybean seeds, the aim of this study was to evaluate seed vigor according to different inoculum potentials. The fungus isolates were inoculated on seeds by the technique of water restriction, through which different inoculum potentials are obtained, corresponding to times of seed exposure of 0, 24, 48, and 96 hours for maize and cotton seeds, and 0, 36, 108, and 144 hours for soybean seeds. The seeds were subjected to germination, electrical conductivity, health, and qPCR tests. Results of the blotter test showed that in most pathosystems, there was a higher incidence of the fungi with an increase in inoculum potential. A decrease in germination percentage was observed in all species as inoculum potential increased, as well as further degradation of seed membranes. The qPCR test confirmed that the most damaged seeds in the tests had higher presence of the pathogens.

scholarly journals Development of Molecular Markers for Detection of Acidovorax citrulli Strains Causing Bacterial Fruit Blotch Disease in Melon

2019 ◽  
Vol 20 (11) ◽  
pp. 2715 ◽  
Author(s):  
Md. Rafiqul Islam ◽  
Mohammad Rashed Hossain ◽  
Hoy-Taek Kim ◽  
Denison Michael Immanuel Jesse ◽  
Md. Abuyusuf ◽  
...  
Keyword(s):  
Infected Leaf ◽  
Bacterial Fruit Blotch ◽  
Early Disease Detection ◽  
Acidovorax Citrulli ◽  
Seed Health ◽  
Field Samples ◽  
Whole Genomes ◽  
Pathogenesis Related ◽  
Primer Sets ◽  
Inoculation Assay

Acidovorax citrulli (A. citrulli) strains cause bacterial fruit blotch (BFB) in cucurbit crops and affect melon significantly. Numerous strains of the bacterium have been isolated from melon hosts globally. Strains that are aggressively virulent towards melon and diagnostic markers for detecting such strains are yet to be identified. Using a cross-inoculation assay, we demonstrated that two Korean strains of A. citrulli, NIHHS15-280 and KACC18782, are highly virulent towards melon but avirulent/mildly virulent to the other cucurbit crops. The whole genomes of three A. citrulli strains isolated from melon and three from watermelon were aligned, allowing the design of three primer sets (AcM13, AcM380, and AcM797) that are specific to melon host strains, from three pathogenesis-related genes. These primers successfully detected the target strain NIHHS15-280 in polymerase chain reaction (PCR) assays from a very low concentration of bacterial gDNA. They were also effective in detecting the target strains from artificially infected leaf, fruit, and seed washing suspensions, without requiring the extraction of bacterial DNA. This is the first report of PCR-based markers that offer reliable, sensitive, and rapid detection of strains of A. citrulli causing BFB in melon. These markers may also be useful in early disease detection in the field samples, in seed health tests, and for international quarantine purposes.

scholarly journals Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites

2018 ◽  
Vol 6 (9) ◽  
pp. 1577-1580
Author(s):  
Nihal A. Hanafy ◽  
Mohamed S. Badr ◽  
Ghada M. Nasr
Keyword(s):  
Toxoplasma Gondii ◽  
Real Time ◽  
Parasitic Infection ◽  
Clinical Samples ◽  
Molecular Assays ◽  
Low Concentrations ◽  
Target Dna ◽  
Primer Sets ◽  
Amplification Failure ◽  
Reference Samples

BACKGROUND: Toxoplasma gondii is a common parasitic infection of humans. Infection is usually mild. Serious complications can occur in pregnant and immunocompromised patients. AIM: The present study aims to investigate the performance of 2 different PCR protocols; real-time quantitative molecular assays (qPCR) and conventional molecular assays (cPCR), using 2 different sets of primers and by using cloned purified Toxoplasma genomic substances to be evaluated as reference samples. METHODS: The target DNA was provided in 8 different quantities. RESULTS: Amplification failure was reported only with the cPCR in samples of low concentrations using both primer sets. Quantitative PCR detected the 8 different dilutions of the purified Toxoplasma gondii using the 2 sets of primers while cPCR was sensitive to detect only 6 different dilutions. CONCLUSION: Generally real-time quantitative molecular assays, is easy to use method compared to conventional PCR assay and produces more reliable results within only one hour time but still the possible application of qPCRs in routine diagnosis necessitates analysis of a large number of clinical samples in further studies to make the proper choice.

scholarly journals Detection of smut mycelia in apical meristems of sugarcane buds

1969 ◽  
Vol 75 (3) ◽  
pp. 281-286
Author(s):  
Rodrigo Echávez-Badel
Keyword(s):  
Apical Meristem ◽  
Staining Method ◽  
Saccharum Officinarum ◽  
Seed Certification ◽  
Apical Meristems ◽  
Sugarcane Varieties ◽  
Certification Programs ◽  
Ustilago Scitaminea ◽  
Fungicide Efficacy ◽  
Nursery Field

Two tests were conducted to detect the fungal mycelia of Ustilago scitaminea Syd. in apical meristems of sugarcane (Saccharum officinarum L.) buds. In the first test six varieties were selected from infected sugarcane fields, and in the second test three varieties obtained from a nursery field were artificially inoculated with the fungus. Sugarcane plants artificially inoculated with U. scitaminea were used as checks in the first test. Growing points were removed from the plant cane and stained for 4 to 18 h by using Sinha's technique. Microscopic observations indicated the absence and presence of smut mycelia in the apical meristem buds of healthy and infected sugarcane varieties. The staining method of growing point nodal buds can be useful for indicating fungicide efficacy in controlling sugarcane smut, and for sugarcane seed certification programs.

Seed Health Testing and Seed Certification

2020 ◽  
pp. 795-808
Author(s):  
Atul Kumar ◽  
Ashok Gaur ◽  
Malkhan Singh Gurjar ◽  
Pooja Kumari ◽  
Raj Kiran
Keyword(s):  
Seed Certification ◽  
Seed Health

scholarly journals A comparison of methods for the detection of Ascochyta rabiei in chickpea seeds

2019 ◽  
Vol 45 (2) ◽  
pp. 197-199
Author(s):  
Francisco José Sautua ◽  
Santiago Agustín Casey ◽  
Raúl Lorenzo Zapata ◽  
María Mercedes Scandiani ◽  
Marcelo Aníbal Carmona
Keyword(s):  
Buenos Aires ◽  
Ascochyta Rabiei ◽  
Buenos Aires Province ◽  
Water Restriction ◽  
Paper Substrate ◽  
Factors Affecting ◽  
Plate Test ◽  
Seed Health ◽  
Paper Method

ABSTRACT Seed health is one of the most important factors affecting the quality of chickpea (Cicer arietinum) seeds. The present study aimed to compare and identify the best incubation methods for detecting Ascochyta rabiei associated with chickpea seeds. Four protocols were compared for their sensitivity in detecting A. rabiei: T1) Incubation on paper substrate or filter paper method (blotter test) without surface disinfection, T2) Blotter test through the water restriction technique, T3) PDA plate test, and T4) MEA plate test. Four independent chickpea seed lots, naturally infected with A. rabiei, were sampled from Córdoba Province and other four were sampled from Buenos Aires Province, Argentina. Each treatment was applied to a total of 400 seeds from each locality for the methods to be comparable. T2 and T3 were statistically more sensitive in detecting A. rabiei-infected seeds from Córdoba. Only these two treatments were repeated for seeds from Buenos Aires Province, and T3 proved to be more sensitive; thus, it is recommended for routine sanitary analysis of chickpea seeds.

scholarly journals Detection of human papillomavirus in cervical cell specimens by hybrid capture and PCR with different primers.

2006 ◽  
Vol 53 (3) ◽  
pp. 603-607 ◽  
Author(s):  
Slawa Szostek ◽  
Malgorzata Klimek ◽  
Barbara Zawilinska ◽  
Janusz Rys ◽  
Jolanta Kope ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
High Risk ◽  
Consensus Primer ◽  
Hybrid Capture ◽  
Hpv Dna ◽  
Cervical Smears ◽  
Capture Assay ◽  
Cervical Cell ◽  
Target Dna ◽  
Primer Sets

The purpose of this study was to compare hybrid capture assay with PCRs using different primers for the L1, E6-E7 regions for the detection of human papillomavirus (HPV) genome. One hundred twenty-five cervical smears with normal (n=42) and abnormal (n=83) cytology were investigated. Those at high-risk for HPV were studied by hybridization antibody capture assay and PCR with the pU-1M/pU-2R primers. Target DNA from the HPV L1 region was amplified by SPF10 primer set and home-PCR with MY09/MY11 primers. The presence of HPV DNA in cervical smears was detected by SPF10 (in 72% of cases), MY09/MY11 (58%), hybrid capture (55%) and pU-1M/pU-2R (39%). Results obtained with the SPF10 and MY09/MY11 consensus primer sets as well as hybrid capture and pU-1M/pU-2R specific for high-risk types differed significantly (chi2, P

scholarly journals Study of cotton seed term sowing territories in areas susceptible to wind erosion in Uzbekistan

2021 ◽  
Vol 258 ◽  
pp. 03016
Author(s):  
Zokirjon Ismatullaev ◽  
Allamurat Khojasov ◽  
Gulnora Akhmedjanova ◽  
Munisa Pulatova ◽  
Gafurjan Isaev
Keyword(s):  
Natural Disasters ◽  
Cotton Seed ◽  
Wind Erosion ◽  
Mineral Fertilizers ◽  
Cotton Yield ◽  
Cotton Seeds

The cotton seeds of Namangan-77, Sultan, S-8290, and S-8286 varieties were sown on May 5 with N-200, P-150, K-100 mineral fertilizers and irrigated at 70-80-70% of LFMC. When the seeds were sown on May 15, 58.4% of cotton was yielded in the first harvest from S-8286 compared to the above varieties. Compared to other varieties, an increase in yield in the first harvest was found to be 1.3-13.5%. Namangan-77, Sultan, S-8290 and S-8286 varieties were sown on May 15 due to wind erosion and other natural disasters. Data on cotton yield of 28.0 q/ha are given.

scholarly journals Enhancing Colorimetric LAMP Amplification Speed and Sensitivity with Guanidine Chloride

2020 ◽  
Author(s):  
Yinhua Zhang ◽  
Guoping Ren ◽  
Jackson Buss ◽  
Andrew J. Barry ◽  
Gregory C. Patton ◽  
...  
Keyword(s):  
Guanidine Hydrochloride ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Simple Method ◽  
Diagnostic Assays ◽  
Dna And Rna ◽  
Target Dna ◽  
Versatile Technique ◽  
Guanidine Chloride ◽  
Primer Sets

AbstractLoop-mediated isothermal amplification (LAMP) is a versatile technique for detection of target DNA and RNA, enabling rapid molecular diagnostic assays with minimal equipment. The global SARS-CoV-2 pandemic has presented an urgent need for new and better diagnostic methods, with colorimetric LAMP utilized in numerous studies for SARS-CoV-2 detection. However, the sensitivity of colorimetric LAMP in early reports has been below that of the standard RT-qPCR tests, and we sought to improve performance. Here we report the use of guanidine hydrochloride and combined primer sets to increase speed and sensitivity in colorimetric LAMP, bringing this simple method up to the standards of sophisticated technique and enabling accurate and high-throughput diagnostics.

scholarly journals Eksplorasi Rafinosa Biji Kapas Sebagai Pengganti Formalin Dalam Pengawetan Ikan [Raffinose Exploration Cotton Seeds Lieu Of Formalin In Preserving Fish]

2019 ◽  
Vol 5 (2) ◽  
pp. 151
Author(s):  
Khairanita K, Pipin Suciati, Kurnia Ayu , Abdul Manan, Moch. Amin Alamsjah
Keyword(s):  
Organic Solvent ◽  
Shelf Life ◽  
Cotton Seed ◽  
Good Condition ◽  
Seed Extract ◽  
Cotton Seeds ◽  
Human Effort ◽  
Fish Remains ◽  
Formalin Preservation

Abstract Preservation is a human effort to enhance durability and shelf life of the fish that the quality of fish remains in good condition. Irregularities in the use of formalin preservation would endanger the health of consumers. Cottonseed is containing compounds of Raffinose, proteins, fats that are cryoprotectants to cells and antimicrobe, that use of cottonseed to suppress use of formalin as a preservative of fish in the community. The method used by dissolving cottonseeds in organic solvents, where in the organic solvent effective to extract the active ingredients in cottonseed. The results showed use of cotton seed extract is 100% equal to 100% use of formalin in maintaining the quality of the fish. This is evidenced by organoleptic tests, pH and bacterial tests. Expected with these results, cottonseed can be a substitute for use of formalin.

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