rDNA-ITS2 polymerase chain reaction assay for the sibling species of Anopheles fluviatilis

Manonmani, A.M.; Townson, Harold; Adeniran, T; Jambulingam, P.; Sahu, Sushanta Kumar; Vijayakumar, T.

doi:10.1016/s0001-706x(00)00154-6

Cited by 59 publications

(50 citation statements)

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“…Species S is found to be highly anthropophilic (91%) and is considered the principal malaria vector in India, while species T and U are generally zoophilic (91%) and are regarded as secondary (in the presence of large populations) or non-vectors [16,17]. Manonmani et al [7] developed an allelespecific PCR assay for the differentiation of the 2 species, referred to as species X and Y, by exploiting differences in the ITS2 gene of rDNA of individuals from Orissa State, India. Subsequent studies showed that cytotaxonomy and ITS2-based methods were 93% concordant for species S and X, and 84% for species T and Y, while no ITS2 gene fragment for species U was reported [18].…”

Section: Discussionmentioning

confidence: 99%

“…The PCR reaction conditions were as recommended by Singh et al [11], except for the amount of primers, which were reduced to 20 pM in 25 ml reactions. Also, the ITS2 region of 23 individuals from different regions, including Minab in Hormozgan Province, was sequenced using the primers and PCR conditions outlined by Manonmani et al [7].…”

Section: Sequencing Of Dnamentioning

confidence: 99%

“…The ITS2 sequence of all individuals was 374 nucleotides in length, free of ambiguities, and identical to all database entries for species T submitted by other authors [5,7,8]. …”

Section: Resutlsmentioning

confidence: 99%

“…fluviatilis complex that were designated as S, T, and U [6]. Later, 2 putative species of X and Y were identified in Orissa State, east-central India, based on differences in the second internal transcribed spacer (ITS2) of ribosomal DNA (rDNA) [7]. Comparison of the ITS2 sequence in Iranian specimens from south and southeastern Iran, including Fars, Sistan va Baluchestan, Kerman, Bushehr, and Hormozgan, showed that they were all homologous to the published ITS2 sequences of species Y (presumably species T) from India [5,8].…”

Section: Introductionmentioning

confidence: 99%

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Molecular Variation and Distribution ofAnopheles fluviatilis(Diptera: Culicidae) Complex in Iran

2010

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Anopheles fluviatilis James (Diptera: Culicidae) is one of the known malaria vectors in south and southeastern Iran. Earlier ITS2 sequences analysis of specimens from Iran demonstrated only a single genotype that was identical to species Y in India, which is also the same as species T. We identified 2 haplotypes in the An. fluviatilis populations of Iran based on differences in nucleotide sequences of D3 domain of the 28S locus of ribosomal DNA (rDNA). Comparison of sequence data from 44 Iranian specimens with those publicly available in the Genbank database showed that all of the 28S-D3 sequences from Kazeroun and Khesht regions in Fars Province were identical to the database entry representing species U in India. In other regions, all the individuals showed heterozygosity at the single nucleotide position, which identifies species U and T. It is argued that the 2 species may co-occur in some regions and hybridize; however, the heterozygosity in the 28S-D3 locus was not reflected in ITS2 sequences and this locus for all individuals was identical to species T. This study shows that in a newly diverged species, like members of An. fluviatilis complex, a single molecular marker may not be sufficiently discriminatory to identify all the taxa over a vast geographical area. In addition, other molecular markers may provide more reliable information for species discrimination.

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Section: Discussionmentioning

confidence: 99%

Section: Sequencing Of Dnamentioning

confidence: 99%

“…The ITS2 sequence of all individuals was 374 nucleotides in length, free of ambiguities, and identical to all database entries for species T submitted by other authors [5,7,8]. …”

Section: Resutlsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Molecular Variation and Distribution ofAnopheles fluviatilis(Diptera: Culicidae) Complex in Iran

2010

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“…Comparatively less is known about the species of the genus Culex which also include species of epidemiological significance. For the PCR based enzymatic amplification of mosquito genome, random as well as gene specific primers has been developed for selected nuclear and mitochondrial gene sequences (Severini et al, 1996;Crabtree et al, 1995;Cornel et al 1996;Proft et al, 1999;Manonmani et al, 2001;Singh et al, 2004;and Li and Wilkerson, 2005). In fact, the major impetus for mosquito genome studies came from WHO/UNDP/World Bank (2003) special programme for research and training in tropical diseases.…”

Section: Introductionmentioning

confidence: 99%

RAPD-PCR based genomic characterization of two populations of Culex quinquefasciatus (Diptera : Culicidae)

Kaura

Kaushal

Bansal

et al. 2009

JANS

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Abstract:The present paper deals with the RAPD-PCR based genomic characterization of Culex quinquefasciatus Say which is a major vector of filariasis in several parts of the Indian subcontinent. One population of the test organism used in the study was procured from Goa (pop.A) while the other (pop.B) was collected from a village Nadasahib (20 kms from Chandigarh). The RAPD-PCR amplification of whole body homogenate of freshly hatched individual specimens was carried out by using three random primers: primer I-5'-GTCCCGACGA -3'; primer II-5' -TGATCCCTGG -3' and primer III-5'-GTGACGTAGG -3'. Primer I produced 5 distinct bands from the DNA of pop. A, whose base composition ranged from 200-1000 bp. Likewise, 7 bands ranging from 130-750 bp and 4 bands ranging from 270-950 bp were generated with primers II and III respectively. In case of pop.B, a total of 8 bands ranging from 200-1000 bp were generated with primer I. Similarly, a total of 6 bands ranging from 250-900 bp with primer II and 4 bands ranging from 180-950 bp with primer III were produced. Based on the band sharing coefficient and the application of Nearest Neighbour Joining (NJ) analysis it was found that primer I was more suitable for detecting genomic differences at the species and generic levels while primer II was ideal for detecting variations in the number of bp in RAPD generated bands among different populations of Cx. quinquefasciatus.

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On the conspecificity ofAnopheles fluviatilis species S withAnopheles minimus species C

et al. 2006

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Anopheles fluviatilis and An. minimus complexes,each comprising of at least three sibling species, are closely related and important malaria vectors in Oriental Region. Recently An. fluviatilis species S, which is a highly efficient malaria vector in India, has been made conspecific with An. minimus species C (senior synonym) on the basis of homology in 335 base pair nucleotide sequence of D3 domain of 28S ribosomal DNA (rDNA). We examined the conspecificity of these two nominal species by obtaining and analysing the DNA sequences of nuclear ribosomal loci internal transcribed spacer 2 (ITS2) and D2-D3 domain of 28S rDNA (28S-D2/D3) from those of An. fluviatilis S and An. minimus C. We found that the sequences of An. fluviatilis S are appreciably different from those of An. minimus C with pair-wise distance (Kimura-2-parametre model)of 3.6 and 0.7%for loci ITS2 and 28S-D2/D3, respectively. Pair-wise distance and phylogenetic analyses using ITS2 sequences of members of Minimus and Fluviatilis Complexes revealed that An. fluviatilis S is distantly related to An. minimus C as compared to any other members of the Fluviatilis Complex. These findings suggest that the two nominal species, An. fluviatilis S and An. minimus C, do not merit synonymy. The study also confirms that the reported species An. fluviatilis X is synonym with species S.

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rDNA-ITS2 polymerase chain reaction assay for the sibling species of Anopheles fluviatilis

Cited by 59 publications

References 8 publications

Molecular Variation and Distribution ofAnopheles fluviatilis(Diptera: Culicidae) Complex in Iran

Molecular Variation and Distribution ofAnopheles fluviatilis(Diptera: Culicidae) Complex in Iran

RAPD-PCR based genomic characterization of two populations of Culex quinquefasciatus (Diptera : Culicidae)

On the conspecificity ofAnopheles fluviatilis species S withAnopheles minimus species C

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