1 s2.0 S1319562X11000234 Main
1 s2.0 S1319562X11000234 Main
REVIEW
a
Molecular Fingerprinting and Biodiversity Unit, Prince Sultan Research Chair for Environment and Wildlife,
College of Sciences, King Saud University, Riyadh, Saudi Arabia
b
Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia
c
Department of Biology, College of Science, Taif University, Taif, Saudi Arabia
KEYWORDS Abstract Use of molecular markers for identification of protected species offers a greater promise
Biodiversity; in the field of conservation biology. The information on genetic diversity of wildlife is necessary to
Conservation; ascertain the genetically deteriorated populations so that better management plans can be estab-
Endangered animals; lished for their conservation. Accurate classification of these threatened species allows understand-
Fingerprinting; ing of the species biology and identification of distinct populations that should be managed with
Molecular markers; utmost care. Molecular markers are versatile tools for identification of populations with genetic cri-
Wildlife sis by comparing genetic diversities that in turn helps to resolve taxonomic uncertainties and to
establish management units within species. The genetic marker analysis also provides sensitive
and useful tools for prevention of illegal hunting and poaching and for more effective implementa-
tion of the laws for protection of the endangered species. This review summarizes various tools of
DNA markers technology for application in molecular diversity analysis with special emphasis on
wildlife conservation.
ª 2011 King Saud University. Production and hosting by Elsevier B.V. All rights reserved.
Contents
1. Mitochondrial DNA markers for wildlife conservation et al., 2003). Shukla et al. (2001) amplified the 12S rRNA gene
using universal primers and then cloned and sequenced the 450
Many of the conservation genetics studies have utilized the se- bp fragment for phylogenetic analysis of endangered species,
quence information of mitochondrial DNA (mtDNA). The Indian muntjac (Muntiacus muntjak). The 12S rRNA gene
mitochondrial genome comprises a circular chromosome of has been used to examine genetic variation in the endangered
DNA. Animal mtDNA ordinarily contains 36 or 37 genes; spur-thighed tortoise (Testudo graeca), an endangered species
two for rRNAs, 22 for tRNAs and 12 or 13 for subunits of from broad distribution range (Alvarez et al., 2000; van der
multimeric proteins of the inner mitochondrial membrane. In Kuyl et al., 2005). The 12S fragment of Testudo graeca was
addition, there is a noncoding sequence termed the control re- found to be somewhat less variable than the D-loop fragment,
gion (CR) due to its role in replication and transcription of a finding compatible with the situation in mammals, where
mtDNA molecules. Exons in the mtDNA circle are tightly rRNA genes evolve slower than synonymous sites and the var-
packed with no spacing introns. Mitochondrial DNA is his- iable parts of the D-loop (Pesole et al., 1999). Recently, Pan-
tone-free, has limited repair ability, and therefore has a rela- dey et al. (2007) have utilized 12S rDNA for molecular
tively high mutation fixation rate (5–10 times that of nuclear identification of Indian leopard, which is an endangered spe-
DNA). Although mtDNA has evolved faster than the nuclear cies except in Central Africa and India.
genome, the rate of evolution is different for different regions Lei et al. (2003) have examined the mitochondrial rRNA
of mtDNA and has been used to examine various phylogenetic genes of Chinese antelopes and observed that average sequence
relationships. Several conserved primers have been developed divergence values for 16S and 12S rRNA genes are 9.9% and
that allow amplification of a number of regions of the mtDNA 6.3% respectively. Their phylogenetic analysis has revealed
molecule in a wide range of species. Moreover, because most that Przewalski’s gazelle is more closely related to Mongolian
cells contain multiple copies of the mtDNA molecule, mtDNA gazelle than Tibetan gazelle suggesting that the critically
sequences can often be obtained from very small amounts of endangered Przewalski’s gazelle should be treated as a species,
tissue containing degraded DNA. The main uses of mtDNA not a subspecies of the Tibetan gazelle, which clearly warrants
sequences in conservation genetics include population structur- more attention from conservationists (Lei et al., 2003). A sin-
ing, resolving taxonomies, establishing interspecific hybridiza- gle base in the 16S rDNA sequences from the endangered spe-
tion and the detection of illegal hunting and poaching of cies Pinna nobilis has been found to be different in all analyzed
endangered animals. However, the selection of appropriate individuals from a single population sample (Chios island) dif-
bioinformatics tool plays an important role for reliable phylo- ferentiating it from the others (Katsares et al., 2008). Mito-
genic inference using mitochondrial markers (Khan et al., chondrial 16S rRNA has been used to elucidate the pattern
2008b). Table 1 provides quick information on numbers of of relationships and systematic status of 4 genera, including
samples and populations, names of taxa and types of mito- 9 species of skates living in the Mediterranean and Black Seas
chondrial markers used in some relevant studies. (Turan, 2008). A heminested PCR assay based on species-spe-
cific polymorphism at the mitochondrial 16S rRNA gene has
1.1. Ribosomal DNA (12S and 16S rDNA) been designed for the identification of seven pecora species
including Blackbuck, Goral, Nilgai, Hog deer, Chital, Sambar
Mitochondrial 12s rDNA is highly conserved and has been ap- and Thamin deer (Guha and Kashyap, 2005). Molecular stud-
plied to illustrate phylogeny of higher categorical levels such as ies on endangered Pecoran have shown lower sequence diver-
in phyla or subphyla. Whereas the 16s rDNA is usually applied sity in 16S rRNA gene as compared to cytochrome b gene,
for phylogenetic studies at mid-categorical levels such as in both between and within species however the 16S rRNA gene
families or genera. It has been postulated that these sequences harbored a larger number of species-specific mutation sites
are useful for inferring moderate to long divergence times (Jan- than cytochrome b gene, suggesting that it could be more use-
czewski et al., 1995). Several investigators have used 12S ful for species identification (Guha et al., 2006). NaNakorn et
rDNA sequences for wildlife forensic biology (Prakash et al., al. (2006) have assessed the level of genetic diversity of criti-
2000; Gurdeep et al., 2004). Molecular phylogeny of elop- cally endangered Mekong giant catfish species using sequences
omorph fishes using 12S rRNA sequences clearly separated of 16S rRNA and detected 4 haplotypes among 16 samples
monophyletic Elopomorpha from Clupeomorpha (Wang from natural populations. These findings may have important
DNA marker technology for wildlife conservation 221
implications for conservation of the Mekong giant catfish, of variation; both ETAS and CSB domains evolve rapidly
especially in designing and implementing artificial breeding whereas the central domain remains taxonomically conserva-
program for restocking purposes (NaNakorn et al., 2006). Re- tive. Iyengar et al. (2006) have compared the CR sequences
cently, Khan et al. (2008a) have suggested the utility of 16S from several captive animals with the sequences for oryx spe-
rRNA segment for molecular phylogeny of oryx at the genus cies and the subsequent phylogenetic analysis of sequence
and possibly species levels. variations revealed a close grouping of Oryx leucoryx with
Oryx gazelle rather than Oryx dammah. Recently, Khan et
1.2. Mitochondrial protein coding genes al. (in press) have observed typical sequence variation in the
CR gene of 23 captive-bred and reintroduced Oryx leucoryx
Compared to 12S and 16S rDNAs, the mitochondrial protein- samples in the form of 7 haplotypes; one of these haplotypes
coding genes evolve much faster and therefore regarded as has been reported earlier while the remaining 6 haplotypes
powerful markers for inferring evolution history in lower cat- are novel and represent different lineages from the founders.
egorical levels such as families, genera, and species. This fea- The CR sequences have been used to investigate the genetic
ture of mtDNA in phylogeny is suitable for resolving status and evolutionary history of the Tibetan gazelle (Zhang
taxonomic uncertainties in conservation genetics. Mitochon- and Jiang, 2006). Onuma et al. (2006) have sequenced the CR
drial cytochrome b sequences have been used to understand of the sun bear (Helarctos malayanus) using 21 DNA samples
the genetic diversity of Tibetan gazelle for better conservation collected from confiscated animals to identify conservation
planning (Zhang and Jiang, 2006). Partial mitochondrial cyto- units including evolutionarily significant units and manage-
chrome b genes of five mammalian specimens and Chromo- ment units. Wu et al. (2006) have used the partial CR (424
Helicase-DNA-binding (CHD) genes of five pheasants have bp) sequences from 47 samples to assess the population struc-
been used to determine whether the specimens were from ille- ture and gene flow among the populations of black muntjac
gally hunted animals (An et al., 2007). Mitochondrial cyto- (Muntiacus crinifrons), a rare species endemic to China. The
chrome b sequence analysis of cooked meat, remnants of the genetic diversity and population structure of the Chinese
bird and the DNA obtained from the wooden chopping block water deer have been investigated by analyzing the 403 bp
revealed that the cooked meat and remnants of the bird were fragment of the mitochondrial DNA CR revealing 18 differ-
of a chicken, but the wooden chopping block was used to chop ent haplotypes in 40 samples (Hu et al., 2006). Idaghdour
the meat of an endangered bird (Gupta et al., 2005). Partial et al. (2004) have sequenced 854 bp of CR from 73 birds to
cytochrome b based molecular phylogenetic trees and genetic describe their population genetic structure of Chlamydotis
distances have indicated that there is considerable genetic undulata, a declining cryptic desert bird whose range extends
divergence between the Korean goral and the Chinese goral, from North Africa to Central Asia. A single CR haplotype
but virtually none between Korean and Russian gorals sug- was identified in New Zealand population, while 17 haplo-
gesting the importance of molecular data for conservation of types were found in Australian populations of brush-tailed
the goral populations of these regions (Min et al., 2004). John- rock-wallabies (Petrogale penicillata), which were introduced
son and O’Brien (1997) have used NADH dehydrogenase sub- to New Zealand from Australia in the early 1870s and have
unit 5 together with 16S rRNA gene for phylogenetic analysis experienced widespread population declines and extinctions
of multiple individuals of 35 species from Felidae family and (Eldridge et al., 2001).
recognized eight significant clusters or species clades that likely
reflect separate monophyletic evolutionary radiations in the 2. Nuclear DNA markers for wildlife conservation
history of this family.
The most commonly used nuclear markers for DNA finger-
1.3. Non-coding or control region sequences printing are random amplified polymorphic DNA (RAPD),
amplified fragment length polymorphism (AFLP) and micor-
The control region (CR) is composed of three domains satellites or short sequence repeats (SSR). Both RAPD and
including ETAS (extended termination associated sequences) AFLP markers do not require prior molecular information
domain, central domain and CSB (conserved sequence block) whereas SSR markers require sequence information to design
domain. Each of the three domains presents a distinct pattern primers. All these markers are widely used for analysis of
222 I.A. Arif et al.
genetic diversity because of large number of loci, which can be population divergence and genetic variation within and be-
screened simultaneously. The differentiating power of these tween two populations of endangered Pampas deer. They dis-
markers is in the following order: SSR > AFLP > RAPD. played substantial genetic variation with all animals possessing
Both RAPD and AFLP have two alleles per locus and there- unique RAPD phenotypes over 105 polymorphic bands pro-
fore considered as dominant markers (presence or absence of duced by 15 primers suggesting the relevance of these findings
a band). In contrast, SSRs are able to recognize both the alleles for developing management guidelines for its conservation
(co-dominant marker) and can easily differentiate between (Rodrigues et al., 2007). Maciuszonek et al. (2005) have dem-
homozygotes and heterozygotes. The application of onstrated the application of population-specific RADP3 mark-
nuclear DNA markers in wildlife fingerprinting is summarized ers by resolving genetic group specific bands for four
in Table 2. indigenous Polish goose breeds, suggesting endangered geese
to be kept as separate flocks for their preservation. The estima-
2.1. Random amplified polymorphic DNA tion of genetic diversity in an Iberian imperial eagle popula-
tion, one of the most threatened bird species in the world,
Random amplified polymorphic DNA (RAPD) markers can has been carried out using 45 arbitrarily primers that amplified
be amplified relatively easily using standard protocol. There 614 loci (59.7% being polymorphic) in 25 individual eagles
is no requirement for any prior genetic information about (Padilla et al., 2000). This RAPD method has revealed a high
the species in question and the method is relatively simple level of heterozygosity in this species while the genetic dis-
and inexpensive. A more serious drawback of RAPDs is that tances estimated could serve to establish more adequate mat-
the low stringency PCR required in the procedure results in ing to preserve genetic variability (Padilla et al., 2000).
a high genotyping error rate and in lower genotyping repro- RAPD analysis has been used to determine the genetic diver-
ducibility compared to single locus markers. Chelomina et al. sity and the population structure of the endangered Blanca
(1999) have used RAPD to examine genetic polymorphism in Cacerena bovine breed on the basis of 1048 loci produced by
the Far-Eastern leopard subspecies and differented wild-living 71 primers (Parejo et al., 2002). These findings could be useful
animals from those kept in captivity; the genetic diversity was to plan more adequate mating in order to maintain the genetic
found to be lower in animals from the zoo (D=0.058) com- diversity and to improve the efficiency of conservation for this
pared to natural population (D=0.137). Neveu et al. (1998) breed (Parejo et al., 2002).
have compared the genetic diversity in four captive groups of
Microcebus murinus with wild mouse lemurs using 5 primers 2.2. Amplified fragment length polymorphism
to amplify 98 different loci. They showed that the captive
groups had lost genetic information with respect to the wild Amplified fragment length polymorphism (AFLP) markers are
sample; the loss of genetic diversity in captive animal groups becoming one of the most popular tools for genetic analysis in
varied according to their number of founders and/or the man- the fields of evolutionary genetics and conservation of genetic
agement of their captive reproduction (Neveu et al., 1998). The resources. These markers are restriction fragments that are
RAPD analysis using 5 decamer primers to test 52 polymor- amplified by PCR following the addition of adapters to the
phic loci of fifteen closed broodstock lines of the Pacific white fragments. Both RAPD and AFLP are multi-locus systems
shrimp revealed that the genetic variation loss probably related making use of primers that simultaneously bind to many dif-
to bottleneck effects and inbreeding (Freitas et al., 2007). ferent parts of a genome resulting in the amplification of many
Rodrigues et al. (2007) have used RAPD markers to analyze loci at the same time. However, the high stringency PCR used
in the AFLP protocol amplifies markers that are more reliable broad-scale genetic structures in snake populations and can
than RAPD and minisatellites. Although per marker informa- provide important inputs into conservation initiatives of focal
tion in dominant and biallelic AFLP markers is low as com- taxa however microsatellites are superior for detecting struc-
pared to microsatellites, the generation of a large number of ture at limited spatial scales (Lougheed et al., 2000). Yue et
markers outweighs this limitation of AFLP. Zenger et al. al. (2004) have applied microsatellite and AFLP markers for
(2006) have used AFLP to evaluate genetic diversity in the monitoring of genetic diversity of three Asian arowana, a
endangered sand tiger shark (Carcharodon taurus) and the highly endangered fish species. Kawka et al. (2007) assessed
great white shark (Carcharodon carcharias); both the species the genetic variability within and among the 3 ostrich breeds
displayed relatively high levels of allelic diversity with a total and also evaluated the genetic distance between them using
of 59 and 78 polymorphic loci respectively. Giannasi et al. microsatellite markers. Chan et al. (2008) have developed 10
(2001) have investigated the use of AFLP to aid determination microsatellite markers using feathers as DNA source from
of the species tree for 24 specimens of a medically important kakerori suggesting the usefulness of isolated loci in studying
snake and suggested that AFLP may prove a valuable aid in population genetics of this endangered forest bird. Banhos et
determining species trees at fine taxonomic levels to facilitate al. (2008) have characterized the microsatellite loci for the con-
the incorporation of molecular data into such activities as anti- servation and management of the Neotropical harpy eagle in
venom production and conservation management. Inbreeding view of providing an excellent set of molecular tools for the
estimates based on both AFLP and microsatellite markers conservation and management of wild and captive harpy ea-
using empirical data from 179 wild and captive-bred old-field gles. Li et al. (2007) have demonstrated the usefulness of the
mice have been found to correlate strongly with pedigree-based two multiplexed microsatellite systems for genetic diversity
inbreeding coefficients suggesting AFLP markers as a valuable studies on two populations of black tiger shrimp. Multiplexing
tool for estimating inbreeding coefficient in natural popula- of microsatellite markers can be performed either using a sin-
tions and for examining correlations between heterozygosity gle dye labeling (if amplicons are of different size) or different
and fitness (Dasmahapatra et al., 2008). Lucchini (2003) have dye labeling (if amplicons are of same size).
compared the multi-locus AFLP results with single-locus
markers (microsatellites) and haploid organellar marker 3. Conclusion
(mtDNA) sequences and suggested that the AFLP technique
could be very useful in a wide range of conservation studies. Molecular markers play an important role in estimating the
relatedness between the individuals by comparing the geno-
2.3. Microsatellites or simple sequence repeats types at a number of polymorphic loci. Several types of molec-
ular markers are available but none of them can be regarded as
Microsatellites or simple sequence repeats (SSRs) were the first optimal for all applications. Most of the systematic and phylo-
PCR-based markers to become widely used for molecular fin- genetic studies have utilized approaches with mtDNA sequenc-
gerprinting. Microsatellites are short sections of DNA where a ing. However, mtDNA phylogeny represents only the
simple motif, generally 1-5 bp long is repeated up to about 100 geneology of a particular gene that is almost only maternally
times. Microsatellite markers are highly polymorphic, abun- inherited. Therefore additional markers targeting nuclear
dant and fairly evenly distributed throughout eukaryotic gen- DNA, such as RAPD, AFLP or microsatellites need to be used
omes. They are also co-dominant markers as heterozygotes for more accurate interpretation of population genetics, biodi-
can be discriminated from homozygotes because alleles at a versity, phylogeny and forensics.
particular microsatellite locus vary in the number of tandem
repeats and can be differentiated on the basis of the resulting
differences in sequence length. The popularity of these markers References
is due to their ease of amplification by PCR, their co-dominant
nature and their typically high levels of allelic diversity at dif- Alvarez, Y., Mateo, J.A., Andreu, A.C., Diaz-Paniagua, C., Diez, A.,
ferent loci. However, due to complex and tiring methodology, Bautista, J.M., 2000. Mitochondrial DNA haplotyping of Testudo
microsatellite loci have not been identified in the majority of graeca on both continental sides of the Straits of Gibraltar. J.
Hered. 91, 39–41.
species. Some cross-species amplification of microsatellites is
An, J., Lee, M.Y., Min, M.S., Lee, M.H., Lee, H., 2007. A molecular
possible between closely related species but high mutation
genetic approach for species identification of mammals and sex
rates in the sequences flanking the microsatellites severely determination of birds in a forensic case of poaching from South
hamper the designing of conserved primers for universal appli- Korea. Forensic Sci. Int. 167, 59–61.
cation. Therefore, new species-specific microsatellite markers Arif, I.A., Khan, H.A., Shobrak, M., Al Homaidan, A.A., Al Farhan,
have to be isolated whenever a species is studied for the first A.H., Al Sadoon, M., 2010. Measuring the genetic diversity of
time, a process that is both time consuming and expensive. Re- Arabian Oryx using microsatellite markers: implication for captive
cently, Arif et al. (2010) have demonstrated a fair degree of breeding. Gene. Genet. Syst. 85, 141–145.
molecular diversity in captive bred and reintroduced popula- Banhos, A., Hrbek, T., Gravena, W., Sanaiotti, T., Farias, I.P., 2008.
tion of Arabian oryx using an array of seven microsatellite Genomic resources for the conservation and management of the
harpy eagle (Harpia harpyja, Falconiformes, Accipitridae). Genet.
loci. Ryberg et al. (2002) have used microsatellite markers to
Mol. Biol. 31, 146–154.
investigate patterns of genetic variation within and between
Bowland, A.E., Bishop, K.S., Taylor, P.J., et al., 2001. Estimation and
populations of alligators distributed at coastal and inland management of genetic diversity in small populations of plains
localities in Texas. Lougheed et al. (2000) have compared the zebra (Equus quagga) in KwaZulu-Natal, South Africa. Biochem.
genetic differentiation among populations of the threatened Syst. Ecol. 29, 563–583.
massasauga rattlesnake using microsatellites and RAPD. Both Chan, C.H., Zhao, Y., Cheung, M.Y., Chambers, G.K., 2008.
types of markers have been found to be suitable for defining Isolation and characterization of microsatellites in the kakerori
224 I.A. Arif et al.
(Pomarea dimidiata) using feathers as source of DNA. Conserv. Khan, H.A., Arif, I.A., Al Farhan, A.H., Al Homaidan, A.A., 2008a.
Genet. 9, 1067–1070. Phylogenetic analysis of oryx species using partial sequences of
Chang, H., Ke, Y.Y., Su, Y.J., Zhang, G.P., Zhu, S.J., 2002. A study mitochondrial rRNA genes. Genet. Mol. Res. 7, 1150–1155.
of the wild and captive green peafowl(Pavo muticus) by random- Khan, H.A., Arif, I.A., Bahkali, A.H., Al Farhan, A.H., Al Homa-
amplified polymorphic DNA. Yi Chuan 24, 271–274. idan, A.A., 2008b. Bayesian, maximum parsimony and UPGMA
Chelomina, G.N., Spiridonova, L.N., Kozyrenko, M.M., et al., 1999. models for inferring the phylogenies of antelopes using mitochon-
Use of RAPD-PCR-analysis of cellular DNA for the evaluation of drial markers. Evol. Bioinform. 4, 263–270.
genetic polymorphism and subspecies diagnostics of the Far Khan, H.A., Arif, I.A., Shobrak, M., Al Homaidan, A.A., Al Farhan,
Eastern leopard Panthera pardus orientalis. Genetika 35, 681– A.H., Al Sadoon, M., in press. Application of mitochondrial genes
687. sequences for measuring the genetic diversity of Arabian oryx.
Dasmahapatra, K.K., Lacy, R.C., Amos, W., 2008. Estimating levels Gene. Genet. Syst.
of inbreeding using AFLP markers. Heredity 100, 286–295. Lei, R., Jian, Z., Hu, Z., Yang, W., 2003. Phylogenetic relationships of
Eldridge, M.D., Browning, T.L., Close, R.L., 2001. Provenance of a Chinese antelopes (sub family Antilopinae) based on mitochondrial
New Zealand brush-tailed rock-wallaby (Petrogale penicillata) Ribosomal RNA gene sequences. J. Zool. 261, 227–237.
population determined by mitochondrial DNA sequence analysis. Li, Y., Wongprasert, K., Shekhar, M., et al., 2007. Development of
Mol. Ecol. 10, 2561–2567. two microsatellite multiplex systems for black tiger shrimp Penaeus
Fang, S., Wan, Q., 2002. A genetic fingerprinting test for identifying monodon and its application in genetic diversity study for two
carcasses of protected deer species in China. Biol. Conserv. 103, populations. Aquaculture 266, 279–288.
371–373. Lougheed, S.C., Gibbs, H.L., Prior, K.A., Weatherhead, P.J., 2000. A
Freitas, P.D., Calgaro, M.R., Galetti Jr, P.M., 2007. Genetic diversity comparison of RAPD versus microsatellite DNA markers in
within and between broodstocks of the white shrimp Litopenaeus population studies of the massasauga rattlesnake. J. Hered. 91,
vannamei (Boone, 1931) (Decapoda, Penaeidae) and its implication 458–463.
for the gene pool conservation. Braz. J. Biol. 67, 939–943. Lucchini, V., 2003. AFLP: a useful tool for biodiversity conservation
Giannasi, N., Thorpe, R.S., Malhotra, A., 2001. The use of amplified and management. C. R. Biol. 326, S43–S48.
fragment length polymorphism in determining species trees at fine Maciuszonek, A., Grajewski, B., Bednarczyk, M., 2005. RAPD-PCR
taxonomic levels: analysis of a medically important snake, Tri- analysis of various goose populations. Folia Biol. 53, 83–85.
meresurus albolabris. Mol. Ecol. 10, 419–426. Masembe, C., Muwanika, V.B., Nyakaana, S., Arclander, P., Siegis-
Guha, S., Kashyap, V.K., 2005. Development of novel heminested mund, H.R., 2006. Three genetically divergent lineages of the Oryx
PCR assays based on mitochondrial 16s rRNA gene for identifi- in eastern Africa: evidence for an ancient introgressive hybridiza-
cation of seven pecora species. BMC Genet. 6, 42. tion. Conserv. Genet. 7, 551–562.
Guha, S., Goyal, S.P., Kashyap, V.K., 2006. Genomic variation in the Min, M.S., Okumura, H., Jo, D.J., et al., 2004. Molecular phyloge-
mitochondrially encoded cytochrome b (MT-CYB) and 16S rRNA netic status of the Korean goral and Japanese serow based on
(MT-RNR2) genes: characterization of eight endangered Pecoran partial sequences of the mitochondrial cytochrome b gene. Mol.
species. Anim. Genet. 37, 262–265. Cells 17, 365–372.
Gupta, S.K., Verma, S.K., Singh, L., 2005. Molecular insight into a NaNakorn U, Sukmanomon S, Nakajima M, et al., 2006. MtDNA
wildlife crime: the case of a peafowl slaughter. Forensic Sci. Int. diversity of the critically endangered Mekong giant catfish (Pang-
154, 214–217. asianodon gigas Chevey, 1913) and closely related species: impli-
Gurdeep, R. et al., 2004. Species determination and authentication of cations for conservation. Anim. Genet. 9, 483-494.
meat samples by mitochondrial 12S RRNA gene sequence analysis Neveu, H., Hafen, T., Zimmermann, E., Rumpler, Y., 1998. Compar-
and conformation-sensitive gel electrophoresis. Curr. Sci. 87, 1278– ison of the genetic diversity of wild and captive groups of
1281. Microcebus murinus using the random amplified polymorphic
Hu, J., Fang, S.G., Wan, Q.H., 2006. Genetic diversity of Chinese DNA method. Folia Primatol. 69, 127–135.
water deer (Hydropotes inermis inermis): implications for conser- Onuma, M., Suzuki, M., Ohtaishi, N., 2006. Possible conservation
vation. Biochem. Genet. 44, 161–172. units of the sun bear Helarctos malayanus) in Sarawak based on
Idaghdour, Y., Broderick, D., Korrida, A., Chbel, F., 2004. Mito- variation of mtDNA control region. Jpn. J. Vet. Res. 54, 135–139.
chondrial control region diversity of the houbara bustard Chlam- Padilla, J.A., Martı́nez-Trancón, M., Rabasco, A., Parejo, J.C.,
ydotis undulata complex and genetic structure along the Atlantic Sansinforiano, M.E., Guijo, M.I., 2000. Genetic variability in the
seaboard of North Africa. Mol. Ecol. 13, 43–54. Iberian imperial eagle (Aquila adalberti) demonstrated by RAPD
Iyengar, A., Diniz, F.M., Gilbert, T., Woodfine, T., Knowles, J., analysis. J. Hered. 91, 495–499.
Maclean, N., 2006. Structure and evolution of the mitochondrial Pandey, P.K., Dhotre, D.P., Dharne, M.S., et al., 2007. Evaluation of
control region in oryx. Mol. Phylogenet. Evol. 40, 305–314. mitochondrial 12S rRNA gene in the identification of Panthera
Iyengar, A., Gilbert, T., Woodfine, T., Knowles, J.M., et al., 2007. pardus fusca (Meyer, 1794) from field-collected scat samples in the
Remnants of ancient genetic diversity preserved within captive Western Ghats, Maharashtra, India. Curr. Sci. 92, 1129–1133.
groups of scimitar-horned oryx (Oryx dammah). Mol. Ecol. 16, Parejo, J.C., Padilla, J.A., Rabasco, A., Sansinforiano, M.E., Marti-
2436–2449. nez-Trancón, M., 2002. Population structure in the endangered
Janczewski, D.N., Modi, W.S., Stephens, J.C., O’Brain, S.J., 1995. Blanca Cacereña bovine breed demonstrated by RAPD analyses.
Molecular evaluation of mitochondrial 12S RNA and cytochrome Genes. Genet. Syst. 77, 51–58.
b sequences in the Pantherine lineage of Felidae. Mol. Biol. Evol. Pesole, G., Gissi, C., De Chirico, A., Saccone, C., 1999. Nucleotide
12, 690–707. substitution rate of mammalian mitochondrial genomes. J. Mol.
Johnson, W.E., O’Brien, S.J., 1997. Phylogenetic reconstruction of the Evol. 48, 427–434.
Felidae using 16S rRNA and NADH-5 mitochondrial genes. J. Pitra, C., Hansen, A.J., Lieckfeldt, D., Arctander, P., 2002. An
Mol. Evol. 44, S98–S116. exceptional case of historical outbreeding in African sable antelope
Katsares, V., Tsiora, A., Galinou-Mitsoudi, S., Imsiridou, A., 2008. populations. Mol. Ecol. 11, 1197–1208.
Genetic structure of the endangered species Pinna nobilis (Mollusca: Prakash, S., Patole, M.S., Ghumatkar, S.V., Nadode, S.D., Shinde,
Bivalvia) inferred from mtDNA sequences. Biologia 63, 412–417. B.M., Shouche, Y.S., 2000. Mitochondrial 12S rRNA sequence
Kawka, M., Horbańczuk, J.O., Sacharczuk, M., et al., 2007. Genetic analysis in wildlife forensics. Curr. Sci. 78, 1239–1241.
characteristics of the ostrich population using molecular methods. Rodrigues, F.P., Garcia, J.F., Ramos, P.R., Bortolozzi, J., Duarte,
Poult. Sci. 86, 277–281. J.M., 2007. Genetic diversity of two Brazilian populations of the
DNA marker technology for wildlife conservation 225
Pampas deer (Ozotoceros bezoarticus, Linnaeus 1758). Braz. J. Wang, W., Chen, L., Yang, P., Hou, L., He, C., Gu, Z., Liu, Z., 2007.
Biol. 67, 805–811. Assessing genetic diversity of populations of topmouth culter
Ryberg, W.A., Fitzgerald, L.A., Honeycutt, R.L., Cathey, J.C., 2002. (Culter alburnus) in China using AFLP markers. Biochem. System.
Genetic relationships of American alligator populations distributed Ecol. 35, 662–669.
across different ecological and geographic scales. J. Exp. Zool. 294, Wu, H.L., Wan, Q.H., Fang, S.G., 2006. Population structure and
325–333. gene flow among wild populations of the black muntjac (Muntiacus
Scott, I.A.W., Hayes, C.M., Keogh, J.S., Webb, J.K., 2001. Isolation crinifrons) based on mitochondrial DNA control region sequences.
and characterization of novel microsatellite markers from the Zool. Sci. 23, 333–340.
Australian tiger snakes (Elapidae: Notechis) and amplification in Ye, X., Zhu, J., Velleman, S.G., Bacon, W.L., Nestor, K.E., 1998.
the closely related genus Hoplocephalus. Mol. Ecol. Note 1, 117– Measurement of genetic variation within and between Japanese
119. quail lines using DNA fingerprinting. Poult. Sci. 77, 1755–1758.
Shankaranarayanan, P., Banerjee, M., Kacker, R.K., Aggarwal, R.K., Yu, Y., Lian, L.S., Wen, J.K., Shi, X.W., Zhu, F.X., Nie, L., Zhang,
Singh, L., 1997. Genetic variation in Asiatic lions and Indian tigers. Y.P., 2004. Genetic diversity and relationship of Yunnan native
Electrophoresis 18, 1693–1700. cattle breeds and introduced beef cattle breeds. Biochem. Genet. 42,
Shukla, M.S., Pidiyar, V., Bhave, N.A., Patole, M.S., Shouche, Y.S., 1–9.
2001. PCR amplification and sequencing of mitochondrial 12S Yue, G.H., Li, Y., Lim, L.C., Orban, L., 2004. Monitoring the genetic
rRNA gene fragment from Muntiacus muntjak (Indian muntjac). diversity of three Asian arowana (Scleropages formosus) captive
Curr. Sci. 80, 617–618. stocks using AFLP and microsatellites. Aquaculture 237, 89–
Turan, C., 2008. Molecular Systematic Analyses of Mediterranean 102.
Skates (Rajiformes). Turk. J. Zool. 32, 1–6. Zenger, K.R., Stow, A.J., Peddemors, V., Briscoe, D.A., Harcourt,
van der Kuyl, A.C., Ballasina, D.L.P., Zorgdrager, F., 2005. R.G., 2006. Widespread utility of highly informative AFLP
Mitochondrial haplotype diversity in the tortoise species Testudo molecular markers across divergent shark species. J. Hered. 97,
graeca from North Africa and the Middle East. BMC Evol. Biol. 5, 607–611.
29. Zhang, F., Jiang, Z., 2006. Mitochondrial phylogeography and genetic
Wang, C.H., Kuo, C.H., Mok, H.K., Lee, S.C., 2003. Molecular diversity of Tibetan gazelle (Procapra picticaudata): implications
phylogeny of elopomorph fishes inferred from mitochondrial 12S for conservation. Mol. Phylogenet. Evol. 41, 313–321.
ribosomal RNA sequences. Zool. Scrip. 32, 231–241.