Detection of rice tungro bacilliform virus in field and glasshouse samples from India using the polymerase chain reaction

1996 ◽  
Vol 58 (1-2) ◽  
pp. 53-58 ◽  
Author(s):  
Indranil Dasgupta ◽  
Bijan K. Das ◽  
Partha S. Nath ◽  
Sankar Mukhopadhyay ◽  
F.R. Niazi ◽  
...  
Plant Disease ◽  
2015 ◽  
Vol 99 (11) ◽  
pp. 1483-1487 ◽  
Author(s):  
Shu Li ◽  
Weijia Hao ◽  
Guanghua Lu ◽  
Jilei Huang ◽  
Chuanhe Liu ◽  
...  

Rice orange leaf disease (ROLD) is caused by rice orange leaf phytoplasma (ROLP) and occurs sporadically in rice-growing areas in countries of eastern and southeastern Asia. ROLD caused severe damage to rice production in South China in the 1980s. Although its impact subsequently declined in South China, it has reemerged as a serious threat recently. Our study showed that ROLD occurrence varies in different seasons and fields. It was more severe in summer-grown crops (from July to October) than in spring-grown crops (from March to July). In most fields, the incidence was less than 10%, and diseased plants were scattered throughout the fields. In 20% of fields, the incidence was between 10 and 30%. In some fields, over 90% of plants were affected, causing crop failure. Typical symptoms of ROLD include orange-colored leaves and poor growth. Diseased plants were determined as positive for ROLP but negative for Rice tungro bacilliform virus, Rice tungro spherical virus, and Rice transitory yellowing virus through polymerase chain reaction and reverse-transcription polymerase chain reaction. Phytoplasma bodies but not virus-like particles were observed by electron microscopy in phloem tissue of diseased leaves. The leafhopper Inazuma dorsalis, previously identified as the unique vector for ROLP, was rare in the affected fields. Another leafhopper, Nephotettix cincticeps, previously considered a nonvector for this phytoplasma, was very common. Transmission tests revealed that this insect could also transmit ROLP; therefore, it might represent a new vector responsible for the recent incidence of ROLD.


Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  

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.


2006 ◽  
Vol 175 (4S) ◽  
pp. 485-486
Author(s):  
Sabarinath B. Nair ◽  
Christodoulos Pipinikas ◽  
Roger Kirby ◽  
Nick Carter ◽  
Christiane Fenske

1991 ◽  
Vol 66 (04) ◽  
pp. 500-504 ◽  
Author(s):  
H Peretz ◽  
U Seligsohn ◽  
E Zwang ◽  
B S Coller ◽  
P J Newman

SummarySevere Glanzmann's thrombasthenia is relatively frequent in Iraqi-Jews and Arabs residing in Israel. We have recently described the mutations responsible for the disease in Iraqi-Jews – an 11 base pair deletion in exon 12 of the glycoprotein IIIa gene, and in Arabs – a 13 base pair deletion at the AG acceptor splice site of exon 4 on the glycoprotein IIb gene. In this communication we show that the Iraqi-Jewish mutation can be identified directly by polymerase chain reaction and gel electrophoresis. With specially designed oligonucleotide primers encompassing the mutation site, an 80 base pair segment amplified in healthy controls was clearly distinguished from the 69 base pair segment produced in patients. Patients from 11 unrelated Iraqi-Jewish families had the same mutation. The Arab mutation was identified by first amplifying a DNA segment consisting of 312 base pairs in controls and of 299 base pairs in patients, and then digestion by a restriction enzyme Stu-1, which recognizes a site that is absent in the mutant gene. In controls the 312 bp segment was digested into 235 and 77 bp fragments, while in patients there was no change in the size of the amplified 299 bp segment. The mutation was found in patients from 3 out of 5 unrelated Arab families. Both Iraqi-Jewish and Arab mutations were detectable in DNA extracted from blood and urine samples. The described simple methods of identifying the mutations should be useful for detection of the numerous potential carriers among the affected kindreds and for prenatal diagnosis using DNA extracted from chorionic villi samples.


2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR

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.


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