ISSN 10227954, Russian Journal of Genetics, 2015, Vol. 51, No. 1, pp. 116–120. © Pleiades Publishing, Inc., 2015.
Rootknot nematodes of the genus
causing considerable yield loss every year, are major
agriculture pests of a wide host range and
is possibly the most damaging plant pathogen .
Their worldwide distribution raises question about its
route of introduction and the genetic structure of this
species. Meanwhile, the genetic variability of patho
gen is necessary for developing effective control prac
tices through breeding programs .
Microsatellite is one of the most popular and versa
tile genetic markers at the individual level in biology
because of their abundance, high polymorphism, high
transferability, and codominant inherited character
. However, the useful marker has been poorly inves
for the reason of insufficient
template amount for multiple analysis, which is diffi
cult to gain from such small organisms.
Recently, 15 microsatellites of
been described from an enriched genomic library,
three of which exhibited a significant level of intrapo
pulation polymorphism with 3 to 7 alleles detected .
The most conventional approach, screening enriched
genomic library for microsatellite motifs clone, is so
challenging, timeconsuming and lowefficiency that
optimal results can’t be produced. In addition to
insufficient of 3 polymorphic loci for us to analysis
genetic structure in various organisms, the loci
couldn’t be successfully amplified from the Chinese
populations that we have researched. The completion
The article is published in the original.
The authors contributed equally to this work.
of genome sequence has provided a highthroughput
approach for identifying microsatellite loci . 4880
microsatellite loci were detected according to the
previously, but no information
of availability and polymorphism about these loci was
Based on the genomic resource, we reported here
PCR (Polymerase Chain Reaction) primers to amplify
microsatellite loci in
and then investi
gated the polymorphism of some loci with three pop
ulations from China. In order to ensure sufficient tem
plates for amplifying candidate loci, a wholegenome
amplification (WGA) step was employed during the
We got the
genome sequences from the
website at http://www.inra.fr/meloidogyne incognita.
Microsatellites mining was carried out using software
Msatfinder2.0.9 with the motif criteria that di, tri,
tetra, penta and hexanucleotide microsatellites
repeat at least 8, 5, 5, 5 and 5 times, respectively. Fur
thermore, Primer 3 software, which is implemented in
Msatfinder, was used to design the primer pairs for
microsatellite bearing sequences with the default set
Three nematode populations were respectively col
lected from vegetable growing areas in Hainan (
= 44) and Shandong (
= 48) provinces in
China. We isolated the egg masses from vegetable roots
and each nematode came from different egg masses.
The species identification was based on morphology
and confirmed using a multiplex PCR which could
Development of Polymorphic Microsatellites
, through Screening Predicted
Microsatellite Loci Based on Genome Sequence
G. Wang*, E. F. Li**, Z. C. Mao, and B. Y. Xie
Key Laboratory of Horticultural Crops Genetic Improvement, Ministry of Agriculture, Institute of Vegetables and Flowers,
Chinese Academy of Agricultural Sciences, Beijing 100081, China
email: **email@example.com; *firstname.lastname@example.org
Received June 23, 2014
—Microsatellites are extensively distributed in the eukaryotic genome, and they are widely used for
their high polymorphism and accessibility. The microsatellites in
, a worldwide agriculture pest,
are inadequate for diversity research. A repertoire of 1620 microsatellites appeared appropriate to design
primer as markers were identified based on the
genome. 120 loci were chosen as candidate, from
which 88 microsatellites were characterized. Finally, we found 13 polymorphic microsatellites with 2 to 23 alleles
in a survey of three nematode populations in China, while other positive loci were monomorphic. These new
molecular markers afford to genetic diversity analysis in
population of poorly investigation. Fur
thermore, the predicted microsatellites have potential values for other plant parasitic nematodes.