ISSN 1022-7954, Russian Journal of Genetics, 2009, Vol. 45, No. 7, pp. 826–832. © Pleiades Publishing, Inc., 2009.
India is well known for possessing diverse buffalo
germplasm comprising over 97 million buffaloes,
accounting for more than 60% of world’s buffalo popu-
lation. There are ten well-characterized breeds distrib-
uted mostly in North and Central-West parts of India
along with more than 70% nondescript buffalo popula-
tion in different agro-climatic zones. These breeds/pop-
ulations are well adapted to agro-climatic conditions of
respective breeding tract, contributing not only in terms
of milk but also meat and other by products. Most of
these animals are reared in unorganized dairy sector
and are facing dilution due to introduction of superior
germplasm like Murrah breed in order to augment the
milk production for economic beneﬁts.
Genetic diversity analysis tools like microsatellite
markers can help in characterization of the descript
breeds and populations for their breeding, conservation
programme. These markers being highly polymorphic,
locus speciﬁc and easy to use by PCR ampliﬁcation,
have been used extensively for genome characteriza-
tion and diversity analysis in many species [1, 2]. Due
to lack of information on buffalo microsatellite mark-
ers, heterologous bovine markers have been tried for
genetic characterization of buffalo [3–5].
Nagpuri is a unique breed of buffalo spread out in
Vidarbha region comprising ﬁve districts viz. Nagpur,
The article is published in the original.
Wardha, Yavatmal, Akola and Amravati of Maharashtra
state of India. Fanners prefer to rear this breed mainly
due to its low maintenance cost, efﬁciency of feed con-
version, moderate production and better adaptation to
local climatic conditions. The breed has shown mar-
ginal decline in population due to use of Surti and Mur-
rah semen in its breeding tract . Though detailed
information on its geographical distribution, manage-
ment practices, breed characteristics, performance etc.,
has been collected, by carrying out surveys in native
tract, information regarding genetic characterization
needs to be established using molecular genetic
approach. The present work was envisaged to assess
genetic bottleneck based on mutation drift equilibrium
in Nagpuri buffalo breed using data generated for
25 heterologous bovine microsatellite markers.
MATERIALS AND METHODS
Sampling and DNA isolation.
Blood samples of 48
unrelated animals of Nagpuri buffalo breed were col-
lected randomly from the breeding tract from Maha-
rashtra state of India (Fig. 1). Genomic DNA was iso-
lated from blood samples using standard phenol-chlo-
roform extraction method . After testing quality and
quantity of DNA samples on agarose gel as well as
spectrophotometrically the samples were further used
for PCR ampliﬁcation of microsatellite loci.
Microsatellite loci and PCR-based proﬁling.
of 25 bovine speciﬁc heterologous microsatellite mark-
Genetic Diversity and Bottleneck Analysis of Nagpuri Buffalo
Breed of India Based on Microsatellite Data
R. S. Kataria, S. Sunder, G. Malik, M. Mukesh, P. Kathiravan and B. P. Mishra
DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources,
P.B. 129, GT Road By-Pass, Karnal 132001 (Haryana), India;
Received November 8, 2007; in ﬁnal form, March 12, 2008
—In this study, 25 heterologous bovine microsatellite markers have been used for the assessment of
genetic diversity in Nagpuri buffalo, an important breed of Central India. For this, 48 DNA samples of unrelated
individuals of Nagpuri buffalo were PCR ampliﬁed and microsatellite alleles were resolved in 6% denaturing,
silver stained Urea-PAGE gel. Genotypic status of individuals at each locus was identiﬁed manually and data
analysis carried out using POPGENE software. Observed number of alleles varied from 2 (ILSTS073 locus) to
8 (HEL13 & ILSTS058 loci) with a mean of 5.24 alleles per locus. Moderate level of heterozygosity (0.45) indi-
cated sufﬁcient genetic diversity existing in this buffalo population.
values for the microsatellite loci anal-
ysed, ranged from 0.10 (ILSTS0I9 locus) to 0.81 (ILSTS058 locus) with a mean of 0.53. No shift in the fre-
quency distribution of alleles and a normal L-shaped curve indicated non-existence of any bottleneck in Nag-
puri. The study thus highlights the usefulness of heterologous bovine microsatellite markers to assess the
genetic variability in buffalo breds as well. Also various diversity indices suggest sufﬁcient genetic variability
within Nagpuri buffalo that can be utilized as initial guidelines for future breeding strategies and conservation.