Molecular Plant Breeding 2015, Vol.6, No.16, 1
-
13
1
Research Article
Open
Access
Development of new set of microsatellite markers in cultivated tobacco and their
transferability in other
Nicotiana
spp.
Madhav M.S.
1,2,
, Siva Raju K.
1
, Gaikwad K.
3
, Vishalakshi B.
2
, Murthy T.G.K.
1
, Umakanth B.
2
1. Biotechnology section, Crop Improvement Division, Central Tobacco Research Institute (ICAR-CTRI), Rajahmundry, India
2. Biotechnology section, Crop Improvement Division, Indian Institute of Rice Research Institute (ICAR-IIRR), Hyderabad, India
3. National Research Centre on Plant Biotechnology (NRCPB), New Delhi, India
Corresponding
author
email:
Molecular
Plant
Breeding,
2015,
Vol.6,
No.16
doi:
10.5376/mpb.2015.06.0016
Received:
28
Jul.,
2015
Accepted:
29
Aug.,
2015
Published:
13
Oct.,
2015
Copyright
© 2015 Madhav et al.,
This
is
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open
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article
published
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Creative
Commons
Attribution
License,
which
permits
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use,
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and
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provided
the
original
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is
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Preferred
citation
for
this
article:
Madhav M.S., Siva Raju K., Gaikwad K., Vishalakshi B., Murthy T.G.K. Umakanth B., 2015,
Development of new set of microsatellite markers in cultivated
tobacco and their transferability in other
Nicotiana
spp.,
Molecular
Plant
Breeding,
6(16)
1
-
13 (doi:
Abstract
Scarcity of molecular markers in tobacco has been a limitation, hampering the acceleration of breeding efforts.
Development of microsatellite markers is a prerequisite for mapping, tagging of many useful qualitative and quantitative traits and
also for the generation of saturated linkage map. Use of microsatellite-enriched genomic libraries an efficient and rapid method for
the identification of clones harboring microsatellite motifs leading to the development of microsatellite markers. In the present study,
a total of 111 microsatellite motifs was identified from the enriched library, of which, 70 motifs (which includes perfect and
imperfect repeat) were used for marker development. These newly developed markers could successfully differentiated different
types of tobacco and diverse cultivars of Flue Cured Virginia (FCV) tobacco. The high rate of transferability (95.5% - 100%) of these
microsatellite markers in a wide range of
Nicotiana
species indicated their potential as viable resources in the inter-specific gene
transfer programme. The set of microsatellite markers developed in this study is a valuable addition to the already available DNA
marker resources in tobacco.
Keywords
Tobacco; Functionality; Transferability; Genetic diversity; Microsatellite
1 Background
Tobacco (
Nicotiana tabacum
L) is one of the major
commercial crops, belonging to the family Solanaceae,
which contains many genuses, including
Nicotiana
.
Several efforts are being undertaken to improve this
crop, which could not be accelerated due to the
non-availability of sufficient number of molecular
markers. It is necessary to develop adequate molecular
markers for reducing the time, effort and cost in
breeding programs like gene targeting, selection and
pyramiding. These molecular genetic markers have
become useful tools to provide a relatively unbiased
estimation of genetic diversity in plants. PCR-based
markers like randomly amplified polymorphic DNA
(RAPDs), amplified fragment length polymorphism
(AFLPs) and simple sequence repeats (SSRs) have an
apparent advantage as cultivar descriptors since they
are unaffected by environmental or physiological
factors (Bowditch et al., 1993; Khan et al., 2008).
Among the different types of molecular markers,
SSRs are more abundant, ubiquitous in presence,
hyper variable in nature and exhibit high polymorphism
(Gupta et al., 1996; Senan et al., 2014). Moreover,
they are highly reproducible, easy to apply, less cost,
less laborious and hence used for cultivar identification in
many crop plants such as barley (Russel et al.,
1997;Maniruzzaman et al., 2014), soybean (Song et al.,
1999; Bisen et al., 2015), potato (Barandalla et al.,
2006; Carputo et al., 2013),
Solanum melongena
(Behera et al., 2006), rice (Yadav et al., 2008; Molla et
al., 2015) and jute (Khan et al., 2008). Microsatellites
have been shown to be almost twice as informative as
dominant markers (RAPDs and AFLPs) and much
more informative than RFLPs in soybean (Powell et
al., 1996) and approximately six times more
informative than RAPD (Rajora and Rahman, 2003).
In addition to their use in determining the genomic
structure and organization, these markers are also
useful for the development of improved variety
through marker aided selection. Through
microsatellite markers were reported by few earlier
studies in tobacco (Bindler et al., 2007; 2011;Tong et
al., 2012; Hughes et al., 2014), their number is limited
as compared to other commercial crops, thereby
demanding the development of more markers.
Generally, tobacco cultivars are distinguished by
