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Molecular Phylogenetic Relationship Between and Within the Fruit

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Molecular Phylogenetic Relationship Between and Within the Fruit Journal of American Science, 2011;7(10) http://www.americanscience.org Molecular Phylogenetic Relationship Between and Within the Fruit Bat (Rousettus Aegyptiacus) and the Lesser Tailed Bat (Rhinopoma Hardwickei) Deduced From RAPD-PCR Analysis Ramadan A. M. Ali Department of Zoology, College for Women, Ain Shams University, Cairo, Egypt [email protected] Abstract: The RAPD-PCR in the present study was used to determine the genetic variation within and among two Egyptian bat species, Rousettus aegyptiacus and Rhinopoma hardwickei. The animals were captured from one locality at Giza governorate, Egypt. A total of 39 bands were amplified by the three primers OPAO2, OPAO8 and OPCO3 with an average 13 bands per primer at molecular weights ranged from 1409 to 107 bp. The polymorphic loci between both species were 34 with percentage 87.18 %. The numbers of monomorphic bands in Rousettus aegyptiacus aegyptiacus and Rhinopoma hardwickei arabium were 14 and 9 bands, respectively. The two species are sharing 5 (12.8 %) monomorphic bands. The similarity coefficients value between the two bat species was ranged from 0.353 to 0.500 with an average of 0.404 (40.4%). Dendrogram showed that, the two bats genotypes are separated from each other into two clusters and more variation among members of Rhinopoma hardwickei arabium was observed in comparison to those of Rousettus aegyptiacus aegyptiacus. It is concluded that, the similarity coefficient value between the two bat species indicates that, the two bat species may have the same origin but are not identical and separated into two clusters. [Ramadan A. M. Ali Molecular Phylogenetic Relationship Between and Within the Fruit Bat (Rousettus Aegyptiacus) and the Lesser Tailed Bat (Rhinopoma Hardwickei) Deduced From Rapd-Pcr Analysis] Journal of American Science 2011; 7(10): 678-687].(ISSN: 1545-1003). http://www.americanscience.org. Key Words: Fruit bat (Rousettus aegytiacus); lesser tailed bat (Rhinopoma hardwickei), RAPD-PCR, Phylogenetic Relationship. 1. Introduction Kalunda et al., 1986; Wellenberg et al., 2002; The order Chiroptera readily separated into two Calisher et al., 2006; Reeves et al., 2006; Towner groups (megabats and microbats) on the basis of the et al., 2009) whereas, Rhinopoma hardwickei feeds morphological features, the gross anatomical on insects, and acts as a vector for some fungal characteristics, feeding habits and behavior diseases such as subcutaneous zygomycosis (Anderson, 1902; Sanborn and Hoogstraal, 1955; (Gugnani, 1999). Hoogstraal, 1962; Gaisler et al., 1972; Qumsiyeh, Rousettus aegyptiacus is the only bat species 1985). The Species Rousettus aegyptiacus has two recorded in Egypt that belong to the megabats subspecies Rousettus aegyptiacus aegyptiacus and (Qumsiyeh, 1985). The species Rousettus Rousettus aegyptiacus arabicus. Rousettus aegyptiacus contains several subspecies of which the aegyptiacus aegyptiacus is distributed in Egypt, subspecies Rousettus aegyptiacus aegyptiacus which Lebanon, Jordan, Cyprus, and Turkey and from presented in Egypt (Corbet, 1978; Qumsiyeh, Levant to the middle of Arabia and Africa (Corbet 1985; Wassif, 1995; Benda and Horacek, 1998; 1978; Harrison and Bates, 1991; Bergmans, 1994; Kwiecinski and Griffithsi, 1999). The family Benda and Horacek, 1998; Albayrak et al., 2008) Rhinopomatidae in Egypt contains a single genus whereas the subspecies Rousettus aegyptiacus with two species, Rhinopoma hardwickei and arabicus distributed in Oman, Aden, and occurred Rhinopoma microphyllum (Qumsiyeh, 1985; from the South Arabia to Pakistan. While, the lesser Wassif, 1995). In Egypt, Rhinopoma hardwickei tailed bat, Rhinopoma hardwickei is represented by species has two subspecies; Rhinopoma hardwickei two subspecies Rhinopoma hardwickei arabium and arabium in northern Egypt and Rhinopoma Rhinopoma hardwickei cystops in Egypt (Qumsiyeh, hardwickei cystops in Southern Egypt (Qumsiyeh, 1985; Wassif, 1995). 1985; Wassif, 1995). Rhinopomatidae bear a unique The megabat Rousettus aegyptiacus feeds on a set of morphological plesiomorphies for which they variety of fruits, depending on what is locally were often regarded as the most primitive group of available (Kinzelback, 1986; Harrison and Bates, microchiroptera close to the common ancestor of 1991). Fruit bats, Rousettus spp. are not only a microbats and megabats (Van Valen, 1979; cause of great economic loss in fruit crops but are Qumsiyehi, 1985). also vectors of domestic animals and human The order Chiroptera is an excellent example of transmissible ectoparasites (Parri et al., 1971; how molecular phylogenetics helps in reassessment http://www.americanscience.org 678 [email protected] Journal of American Science, 2011;7(10) http://www.americanscience.org of the taxonomy of a well-established group. Genetic between species (Welsh and McClelland, 1990; data invalidated the traditional subdivision of bats Williams et al., 1993; Yadav and Yadav, 2007) and into two divergent suborders; Megachiroptera and within species (Singh and Sharma, 2002; Microchiroptera. Teeling et al. (2000, 2002 and Hassanien et al., 2004; Abdel-Rahman and Hafez, 2005) provided molecular evidence supporting sister 2007) based on the amplification of variable regions position of one clade of microbats, Rhinolophoidea of a genome. (superfamily) with megabats, Pteropodoidea Many researchers used RAPD-PCR markers to (superfamily). The family Rhinopomatidae is differentiate and determine the genetic variation arranged among superfamily Rhinolophoidea (Van between and within species of bats (Sinclair, 1996; Den Bussche and Hoofer, 2004; Eicki et al., 2005; Emmanuvel and Marimuthu, 2006; Moreiral and Teeling et al., 2005). Rhinopomatidae is the most Morielle-versute, 2006; Karuppudurai et al., enigmatic group of extant bats. It is monotypic 2007; Karuppudurai and Sripathi, 2010). Other family composed of a single genus Rhinopoma. molecular studies such as cytochrome B, Rhinopomatidae were regarded as the most primitive microsatellite, RFLP and DNA sequences had been group of Microchiroptera close to the common carried out to solve evolutionary problems in bats ancestor of microbats and megabats (Hill, 1977; Van (Mindell et al., 1991; Colgan and Flannery, 1995; Valen, 1979; Koopman, 1993; Eick et al., 2005). kirsch et al., 1995; Alvarez et al., 1999; Rossiter et Rhinopomatids are the most primitive clade within al., 1999; Ditchfield, 2000; Juste et al., 2002; superfamily Rhinolophoidea and thus, also the most Hulva and Horacek, 2002; Sebastien et al., 2005; primitive extant clade within the suborder Hulva et al., 2007; Levin, 2008; Goodman et al., Yinpterochiroptera (Teeling, 2005). For all these 2010; Chinnasamy et al., 2011; Wood et al., 2011). reasons, Rhinopomatidae are an extremity attractive The present study was mainly focused on subject for future detailed studies. Hulva et al. clarifying the similarities and divergences within and (2007) suggested that, the taxonomic characteristics among individuals of the Rousettus aegyptiacus of Rhinopomatidae are shared with suborder aegyptiacus and Rhinopoma hardwickei arabium Yangochiroptera and Rhinolophoidea of suborder species commonly present in the local environment Yinpterochiroptera partly resembling the condition by using random amplified polymorphic DNA- in Pteropodidae (suborder Yinpterochiroptera). polymerase chain reaction (RAPD-PCR) technique. The analysis of genetic diversity and relationship between or within different species, 2. Materials and Methods populations and individuals is a central task for Animals many disciplines of biological science. The The present work was carried out on the fruit systematic similarity and diversity between bat, Rousettus aegyptiacus aegyptiacus and the lesser Rousettus aegyptiacus and Rhinopoma hardwickei tailed bat, Rhinopoma hardwickei arabium. species was tackled according to chromosomal Animals were trapped near Abu-Rawash area, Giza studies (Ray-Chaudhuri et al., 1968; Qumsiyeh governorate, Egypt. Liver samples were obtained and Baker, 1985), biochemical data (Juste et al., from 6 animals of both Rousettus aegytiacus 1996 and 1997) and molecular genetics (Alvarez et aegyptiacus and Rhinopoma hardwickei arabium and al., 1999; Juste, 2002; Hulva and Horacek, 2002; stored in liquid nitrogen until DNA extraction. Hulva et al., 2007; Goodman et al., 2010). The techniques of molecular genetic markers DNA extraction have an important potential role in detection of Genomic DNA was extracted from liver tissue. genetic differences among and within species. The The extracted DNA quality and concentration was most common molecular techniques are RAPD-PCR examined with both spectrophotometric analysis and (Randomly Amplified Polymorphic DNA- run in 7.0 % agarose gels. In spectrophotometric Polymerase Chain Reaction), SSR-PCR (Simple analysis, each sample of DNA was calculated by Sequence Repeat-Polymerase Chain Reaction), their optical density values at 260 and 280 nm. RFLP (Amplified Fragment Length Polymorphism), Optical density ratios were evaluated and only good and sequencing of nuclear and mitochondrial genes. quality DNA samples were used in PCR. Erlich and Arnheim (1992) reported that the polymerase chain reaction (PCR) has become one of RAPD-PCR the most widely used techniques of molecular A total of five arbitrary decamer primers from biology because it is rapid, inexpensive and Kits OP-A and OP-C (Operon Technologies, produces useful amounts of DNA from small Alameda, CA, USA) were used for RAPD-PCR quantities
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