Evaluation of genotoxicity of acetamiprid using PCR technique on mosquito genome

The present studies deal with the evaluation of the genotoxic potential of acetamiprid at LD40 on a mosquito Culex quinquefasciatus by adopting polymerase chain reaction technique (PCR). This technique was used for detecting DNA damage by amplifying ribosomal DNA internal transcribed spacer 2 (ITS 2) regions. The amplified products were sequenced and the results of treated and non-treated controls were compared using Clustal W software programme. The results were studied in the form of deletions, additions, transitions and transversions of the bases. The DNA band amplified from control stocks consisted of 444 bases while those from LD40 treated individuals were comprised of 448 bases. The total number of mutations caused in the treated stock was 230 out of which 84 were transitions, 117 transversions, 13 deletions and 16 additions. Thus, it was evident that acetamiprid has a potential to promote gene mutations in the individuals exposed to its semilethal doses.

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Molecular approach to evaluate the genotoxicity of glyphosate (roundup) using mosquito genome

Journal of Applied and Natural Science ◽

10.31018/jans.v2i1.105 ◽

2010 ◽

Vol 2 (1) ◽

pp. 96-101

Author(s):

Mamta Bansal ◽

Asha Chaudhry

Keyword(s):

Polymerase Chain Reaction ◽

Dna Damage ◽

Ribosomal Dna ◽

Broad Spectrum ◽

Internal Transcribed Spacer ◽

Gene Mutations ◽

Molecular Approach ◽

Chain Reaction ◽

Dose Concentration ◽

Polymerase Chain

Glyphosate, an active ingredient in Roundup is a broad spectrum, systemic and non -selective herbicide which is commonly used for eliminating weeds in agriculture and forest landscapes. The present studies deal with the evaluation of the genotoxic potential of Glyphosate with two different dose concentration of LD20 and LD40 on a mosquito Culex quinquefasciatus taken as an experimental model. For this, polymerase chain reaction technique (PCR) was used for detecting DNA damage by amplifying ribosomal DNA internal transcribed spacer 2 (ITS 2) region. The amplified products were sequenced and the results of treated and non-treated controls were compared by using Clustal W software programme. The results were studied in the form of transitions, transversions, deletions and additions of bases. The DNA band amplified from control stocks consisted of 440 bases while thosefrom LD20 and LD40 treated individuals were comprised of 423 and 468 bases respectively. The total number of mutations caused in LD20 treated stock was 205 out of which 68 were transitions, 90 transversions, 32 deletions and 15 additions. In case of LD40 treated individuals, as many as 221 bases had suffered mutations, out of which 66 were transitions, 90 transversions , 12 deletions and 41 additions. In both the cases the rate of transversions was higher than transitions. From these results it was evident that glyphosate has a potential to promote gene mutations in the individuals exposed to its semilethal doses.

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Minimal intraspecific variation in the sequence of the transcribed spacer regions of the ribosomal DNA of lake trout (Salvelinus namaycush)

Genome ◽

10.1139/g94-094 ◽

1994 ◽

Vol 37 (4) ◽

pp. 664-671 ◽

Cited By ~ 11

Author(s):

Lang Zhuo ◽

S. L. Sajdak ◽

R. B. Phillips

Keyword(s):

Polymerase Chain Reaction ◽

Intraspecific Variation ◽

Ribosomal Dna ◽

Internal Transcribed Spacer ◽

Lake Trout ◽

Intergenic Spacer ◽

Coding Region ◽

Chain Reaction ◽

Internal Transcribed Spacer Region ◽

Polymerase Chain

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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Identification of Xiphinema vuittenezi by polymerase chain reaction

Plant Protection Science ◽

10.17221/3120-pps ◽

2010 ◽

Vol 40 (No. 1) ◽

pp. 1-4

Author(s):

S. Kumari ◽

K. Kundu J ◽

J. Polák

Keyword(s):

Polymerase Chain Reaction ◽

Ribosomal Dna ◽

Internal Transcribed Spacer ◽

Nematode Species ◽

Specific Primer ◽

Chain Reaction ◽

Polymerase Chain ◽

Species Specific

So far, the identification of the nematode species Xiphinema vuittenezi relied mainly on time-consuming morphological and morphometrical studies. Therefore, a polymerase chain reaction (PCR) protocol was optimised that both reliably and rapidly identifies X. vuittenezi. The internal transcribed spacer (ITS) species-specific primer of ribosomal DNA gene of X. vuittenezi was used. Nine populations of this species from Central Bohemia were investigated by means of PCR.

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