Methodologies for Plasmopara halstedii Research

Helia ◽  
2019 ◽  
Vol 42 (71) ◽  
pp. 173-186
Author(s):  
Ana Laura Martínez ◽  
Freda Anderson ◽  
Facundo Quiroz ◽  
Antonio Garayalde ◽  
Ignacio Erreguerena ◽  
...  
Keyword(s):  
Expressed Sequence Tags ◽  
High Yield ◽  
Infected Leaf ◽  
Chain Reaction ◽  
Plasmopara Halstedii ◽  
Yield And Quality ◽  
Polymerase Chain ◽  
Expressed Sequence

Abstract The objective of this work was to find practical procedures to overcome methodological drawbacks encountered during studies on sunflower downy mildew. Techniques for recovering living isolates of Plasmopara halstedii from the field and for the preservation of infected leaf samples for further molecular analysis were developed. A Polymerase Chain Reaction (PCR)-based test for the detection of P. halstedii in sunflower leaves and a method to remove azoxystrobin from fungicide-treated seeds are proposed. In situ-inoculations of pre-germinated seeds allowed the recovery of living isolates from the field. Three sample-preservation methods were evaluated (silica, heating and lyophilization) resulting in high yield and quality of the DNA extract. It was detected the presence of the pathogen in symptomless leaves through PCR using molecular markers based on expressed sequence tags. A treatment using sodium hypochlorite is recommended for the removal of azoxystrobin from fungicide treated seeds.

Development of an EST-SSR marker in Panax ginseng

2008 ◽  
Vol 5 (2) ◽  
pp. 175-181 ◽  
Author(s):  
Yang Cheng-Jun ◽  
Wang Jun ◽  
Mu Li-Qiang ◽  
Li Shao-Chen ◽  
Liu Guan-Jun ◽  
...  
Keyword(s):  
Panax Ginseng ◽  
Expressed Sequence Tags ◽  
Genomic Dna ◽  
Ssr Marker ◽  
Panax Quinquefolius ◽  
Chain Reaction ◽  
Acanthopanax Senticosus ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Expressed Sequence

AbstractA total of 791 microsatellites (SSRs) were isolated from 7055 Panax ginseng expressed sequence tags (ESTs). According to primer design criteria, 68 primer pairs for EST-SSR were designed. Under an appropriate polymerase chain reaction (PCR) system, all EST-SSR primer pairs were screened against genomic DNA of Ji'anchangbo and Fusong'ermaya from Panax ginseng, and 43 EST-SSR primer pairs out of the above 68 resulted in PCR products. Then, all 43 pairs were detected in nine P. ginseng, two Panax quinquefolius and two Acanthopanax senticosus cultivars for polymorphisms, and 26 pairs (60.47%) were found to be polymorphic, accounting for 38.23% of the total number of designed primer pairs. These results demonstrate the possibility of developing EST-SSR markers using P. ginseng ESTs.

High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

1996 ◽  
Vol 54 ◽  
pp. 896-897
Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
High Performance ◽  
Detection System ◽  
Direct Methods ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

Pengaruh Jenis Co-Solvent terhadap Rendemen dan Mutu Biodiesel secara Transesterifikasi In Situ Menggunakan Radiasi Gelombang Mikro

2016 ◽  
Vol 10 (2) ◽  
pp. 119-126
Author(s):  
Mahlinda Mahlinda ◽  
Fitriana Djafar
Keyword(s):  
Catalyst Concentration ◽  
Maximum Yield ◽  
Acid Value ◽  
Experimental Result ◽  
In Situ Transesterification ◽  
Yield And Quality ◽  
Solvent Type ◽  
Biodiesel Quality

The main purpose of this research was to observer effect co-solvent type (n-Hexane, chloroform and without co-solvent)  toward yield and quality of biodiesel via in situ transesterification process using microwave irradiation. The process was studied at microwave power 450 watt, reaction time 4 minutes, methanol to seed ratio 25:1 and catalyst concentration 5%. The physicochemical parameters of the biodiesel produced such as viscosity, density and acid value were analysed and compared with the SNI 7182-2012 standard. The experimental result showed the maximum yield biodiesel 78,32% obtained by using co-solvent chloroform.Test result of physicochemical properties (viscosity, density and acid value) of biodiesel products using co solvent n-Hexane, chloroform and without co solvent showed that these products conform to the SNI 7182-2012 standars. The type of co-solvent only affectedon biodiesel yield dan not affected on biodiesel quality (viscosity, density and acid value).  ABSTRAKTujuan penelitian ini adalah untuk mempelajari pengaruh jenis co-solvent (n-Hexane, chloroform dan tanpa co-solvent) terhadap rendemen dan mutu biodiesel secara trasesterifikasi in situ menggunakan radiasi gelombang mikro. Proses dilakukan pada daya gelombang mikro 450 watt, waktu reaksi 4 menit, perbandingan berat metanol terhadap bahan baku 25:1 dan jumlah katalis 5%. Parameter fisiko kimia dari produk biodiesel seperti viskositas, densitas dan angka asam di analisa dan dibandingkan dengan standar SNI 7182-2012 tentang biodiesel. Hasil penelitian menunjukkan rendemen maksimum biodiesel sebesar 78,32% diperoleh dengan menggunakan co-solvent chloroform. Hasil pengujian  karakteristik fisiko kimia (viskositas, densitas dan angka asam) dari produk biodiesel menggunakan co-solvent n-Hexane, chloroform dan tanpa co-solvent menunjukkan bahwa semua parameter ini masih memenuhi standar SNI 1782-2012 tentang biodiesel. Jenis co-solvent hanya berpengaruh pada rendemen biodiesel dan tidak berpengaruh terhadap mutu biodiesel (viskositas, densitas dan bilangan asam).Kata kunci: co-solvent, in situ transesterifikasi, microwave, rendemen, mutu   

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