Citation
Khaledi, Kourosh Jome
(2008)
Molecular genetic characterization of Rusa (cervus timorensis) and sika (cervus nipon) deer Species in Malaysia.
PhD thesis, Universiti Putra Malaysia.
Abstract
The Malaysian livestock industry is an important component of the agricultural
sector providing gainful employment and producing useful animal protein food
to the population. Cattle, buffalo, goat, sheep and swine are the popular
livestock in Malaysia. However, in recent years deer farming for meat, fur and
velvet has become popular as well. Most of the farmed deer are of various
species imported from different sources, such as Australia, New Zealand,
Mauritius, Indonesia, New Caledonia, etc. The genetic background of these
species and populations from different sources are unknown. This study was
conducted to characterize two popular deer species in Malaysia, namely the
rusa deer (Cervus timorensis) and the sika deer (Cervus nippon), using DNA
microsatellite markers.
The use of amelogenin gene primers for sexing of the rusa deer was also
investigated. Random samples of 38 rusa deer from the Deer Breeding Unit of the University Research Park, Universiti Putra Malaysia, and 34 sika deer from
Pusat Ternakan Haiwan, Batu Arang, were used in the study to determine and
compare the genetic structures of the two deer species.
One hundred and twenty five sets of microsatellite primer pairs, which had
been reported to have successfully detected variation in deer, cattle or sheep,
were used in the initial screening. Thirty nine primer pairs produced clear and
reproducible amplification products for rusa and 41 primer pairs for sika.
Twenty one primer pairs were polymorphic for the pooled data. However, only
nine microsatellite loci (23.08%) were polymorphic for rusa and 17 loci
(41.46%) were polymorphic for sika. Of these, only five were common to both
deer species (BMS789, BM888, BL4, BM3628 and NVHRT16). Of the
monomorphic loci, 17 were common to both species. Among the 11 reindeer
microsatellite loci screened, nine loci (81%) were amplified for the pooled
data, but only four loci were common to both species. The two white-tailed
deer microsatellite loci (L35582 and L35583) produced amplification in sika
but only L35583 was amplified in rusa. The 17 common monomorphic loci and
the nine polymorphic loci generated in total 53 and 40 microsatellite markers in
the rusa and sika, respectively. Locus BM2113 was amplified exclusively in
rusa (126 bp), and locus NVHRT34 was amplified only in sika (134 bp).
These loci may be used as unique markers to distinguish the two deer species.
The numbers of observed and effective alleles per polymorphic loci were 2 - 13
and 1.05 - 8.91 for rusa, and 2 - 8 and 1.16 - 5.98 alleles per locus for sika,
respectively. The allele frequencies ranged from 0.01 to 0.97 for rusa, and 0.02 to 0.92 for sika. The sizes of the alleles at the polymorphic loci ranged from
116 to 389 bp in rusa, and 88 to 364 bp in sika. The mean numbers of effective
alleles were 3.08±2.40 and 2.87±1.65 in rusa and sika, respectively. Six loci in
rusa and three loci in sika exhibited rare alleles. The mean observed
heterozygosity in the rusa and sika populations were 0.48 ± 0.35 and 0.51 ±
0.30, respectively. Seven polymorphic loci in rusa and 14 polymorphic loci in
sika exhibited significant (P<0.01) deviations from Hardy-Weinberg
equilibrium. The Hardy-Weinberg disequilibrium may be due to overlapping of
generations and founder effect, especially in the sika deer population. The
combined discrimination power (cDP) of the nine polymorphic loci in rusa was
0.99 and of the seventeen polymorphic loci in sika was 0.99, thus allowing
individual identification. The inbreeding coefficient (FIS) was very low for the
rusa population (0.06), but for the sika population it was 0.26. The mean value
of FST was 0.67 for rusa and 0.47 for sika. The bottleneck analysis suggested
that the rusa population did not experience any recent bottleneck, whereas the
sika population had encountered a genetic bottleneck in the recent past.
Evaluation of intra-interchromosomal linkage disequilibrium between the
alleles suggested significant (P<0.05) linkage between 10 pairs of alleles in
rusa and 12 pairs of alleles in sika. However, none of the allelic pairs were the
same for the two species. The genetic distance within the rusa population was
lower than that within the sika population (0.088 ± 0.001 vs. 0.184 ± 0.001).
The genetic distance between rusa and sika was 0.35. No distinct clustering
was observed for the rusa population. The sika population displayed two major
clusters of 11 and 23 individuals. The larger cluster in turn had two subclusters. The above results show the rusa and sika populations to be genetically different
from each other. High genetic variation exists in both the populations. This
could be due to low inbreeding and no directional selection in these
populations.
Four amelogenin gene primer pairs were used to identify the sexes of the rusa
deer. Three primer pairs, AMEL2, AMGX/Y and AMGX/Y2, exhibited similar
banding patterns for the males and females. The primer pair SE47/48 generated
one band for the females (269 bp) but three bands (223, 269 and 305 bp) for the
males. Therefore, this primer pair is a reliable tool for the identification of the
sexes in the rusa deer.
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Additional Metadata
Item Type: |
Thesis
(PhD)
|
Subject: |
Cervus timorensis - Molecular genetics - Case studies |
Subject: |
Sika deer - Molecular genetics - Case studies |
Call Number: |
FP 2008 21 |
Chairman Supervisor: |
Associate Professor Jothi Malar Panandam, PhD |
Divisions: |
Faculty of Agriculture |
Depositing User: |
Rosmieza Mat Jusoh
|
Date Deposited: |
31 Mar 2010 04:58 |
Last Modified: |
27 May 2013 07:17 |
URI: |
http://psasir.upm.edu.my/id/eprint/4725 |
Statistic Details: |
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