Basic HTML Version
Dong et al., 2011, Preliminary mapping of soybean dominant locus
Hrcs7
conferring resistance to
Cerocospara sojina
race 7, Molecular Plant Breeding Vol.2
No.6 (doi: 10.5376/mpb.2011.02.0006)
40
PCR reactions were composed of 1×PCR Buffer, 2
mmol/L MgCl
2
, 100 μmol/L of each dNTP, 0.4 μmol/L
of each primer, 20 ng~25 ng template DNA, and 1 U
of
Taq
DNA polymerase in a total volume of 20 μL.
PCR was carried out with the conditions 94°C for 30s,
47°C for 30s, 72°C for 30s, return to step 1 35 times,
72°C for 3 min. PCR products were separated on 6%
SDS-polyacrylamide
gel
electrophoresis
and
visualized by silver staining (Tiler et al., 1997)
3.3 SSR and linkage analysis
A total of 600 SSR markers were selected out of 20
genetic
linkage
groups
(http://bldg6.-arsusda.
gov/~pooley/soy/cregan/soymap.htm) every 5 cM~10
cM and tested for DNA amplification. The
polymorphism markers between the ‘Gang 95144-1’
and ‘Gongjiao 9723-6 were screened against two
DNA bulks (resistant and susceptible). The
polymorphism markers between the two DNA bulks
were screened against the 184 F
2
individuals.
The resistance trait and genetic segregation
distribution of 184 F
2
individuals were both detected
by the Chi-square (χ
2
) test for goodness of fit. The
data obtained from the F
2
population of ‘Gang
95144-1’ × ‘Gongjiao 9723-6’ were analyzed using
Mapmaker/Exp v 3.0 (Lander et al., 1987). The
Kosambi map function of Mapmaker/Exp v 3.0 was
used to develop a linkage map and obtain centimorgan
(cM) values. The markers were assigned to linkage
groups by the “group” command with a 3.0 minimum
LOD and 50 cM maximum distance and then arranged
by the “order” command. Basing on the published
map the linkage groups were anchored to soybean
chromosomes (Cregen, 1999).
Authors’contributions
DZM and WSM analyzed the trait phenotype and the genotype data and
drafted the manuscript. LJ worked on the trait phenotype identification. LZ
had the experiment on identification of SSR markers. LZG performed the
field experiment management. All authors read and approved the final
manuscript.
Acknowledgements
We thank Dr. Liang Chen (Biotechenology Centre, Jilin Academy of
Agriculture Science, Chuangchun, China) for critical reading and formatting
of the manuscript, and Professor Pengyin Chen (Department of crop, soil,
and enviromental sciences, University of Arkansas, Arkansas, American) for
help and advice on the experiment .This study was jointly supported by
Chinese National Programs for High Technology Research and
Development(Grant No.2006AA100104-6 and No 2006AA10Z1F1-2)
.
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