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the candidate lines were carried out by using t test,
when the character value went to significant difference
the corresponding t-values will be marked in the upper
right corner.
3.3 Extraction of genomic DNA of rapeseed
We adopted the CTAB methods developed by Li et al
(1994) with slightly modification to extract genomic
DNA from rapeseed leaves. The bulk of leaves mixed
from five similar plant type individuals of each line
were ground. The concentration of dissolved DNA
was examined by 1% agarose gel and adjusted the
concentration of each line.
3.4 Screening of SSR primers and amplification
Using 75 pairs of SSR primers, genomic DNA of
NILs of T78, T84 and ZS9 were amplified to screen
the primer pairs with good polymorphism, appropriate
band size and stable amplification. PCR procedure
was performed following the reference of Long et
al (2009). PCR products were separated by 8%
polyacrylamide gel electrophoresis with 20 V/cm until
the front of the Loading buffer migrated to the other
edge, and then silver staining was carried out.
3.5 Band scoring, statistics and cluster analysis
Bands amplified by SSR markers with clear, sharp and
heavy band were scored and calculated statistically.
Band was assigned to be 1, and no band was assigned
to be 0 based on the relevant position. Unclear band
was assigned to be “-”. Dendrogram was drawn by
cluster analysis using NTSYS software.
Author’s contributions
WHL and MLH are the persons who carried out this
experiment; JQG, HMP, SC, and JFZ participated in some in
lab work and involved the data analysis and in field work; CKQ
conceived the project and designed the experiments as well as
wrote and revised manuscript. All authors had read and agreed
the final text.
Acknowledgements
This research is jointly sponsored by the National 863 Project
(No.2011AA10A104), Supporting program of Jiansu province
(BE2008369) and Independent Agricultural Innovation Program of
Jiangsu Province (cx(09)636) Authors thank for two anonymous
reviewers with their critical comments. In this paper we
mentioned some chemical and reagent suppliers and sequencing
service providers, that doesn't mean we would like to
recommend or endorse the production of theirs.
Reference
Fu S.Z., Qi C.K., and Tang J.H., 1989, Breeding of cytoplasmic male sterile
line MI CMS in
Brassica napus
L., Zuowu Xuebao (Acta Agronomica
Sinica), 15(4): 305-309
Fu T.D., 1999, Rapeseed production and variety improvement in China,
Huazhong Nongye Daxue Xuebao (Journal of Huazhong Agriculture
University), 18(6): 501-504
Fu T.D., 2000, Breeding and utilization of rapeseed hybrid, Hubei Science
and Technology Press, Wuhan, China, pp.11-20
Jean M., Brown G.G., and Landry B.S., 1997, Genetic mapping of nuclear
fertility restorer genes for the ‘Polima’ cytoplasmic male sterility in
canola (
Brassica napus
L.) using DNA markers. Theoretical and Applied
Genetics, 95(3): 321-328 http://dx.doi.org/10.1007/s001220050566
Jean M., Brown G.G., and Landry B.S., 1998, Targeted mapping approaches
to identify DNA markers linked to the Rfp1 restorer gene for the
‘Polima’ CMS of canola (
Brassica napus
L.). Theoretical and Applied
Genetics, 97(3): 431-438 http://dx.doi.org/10.1007/s001220050913
Jin X.W., Wang C.L., Yang Q., Jiang Q.X., Fan Y.L., Liu G.C., and Zhao
K.J., 2007, Breeding of Near-Isogenic line CBB30 and molecular
mapping of xa30(t), a new resistance gene to bacterial blight in rice,
Zhongguo Nongye Kexue (Scientia Agricultura Sinica), 40(6):
1094-1100
Li G., and Quiros C.F., 2001, Sequence-related amplified polymorphism (SRAP),
a new marker system based on a simple PCR reaction: its application to
mapping and gene tagging in Brassica, Theoretical and Applied Genetics,
103(2): 455-461 http://dx.doi.org/10.1007/s001220100570
Li J., and Shen B.Z, 1994, An effective procedure for extractive total DNA
in rape, Huazhong Nongye Daxue Xuebao (Journal of Huazhong
Agriculture University), 13(5): 521-523
Liu L.K., Hou N., Liu J.C., Liu G.Q., and Liu C.G., 2006, Screening of the
near-isogenic lines (NILs) of fertility restorer genes for D^2-type
cytoplasmic male sterility (CMS) in wheat and molecular
characterization of the genetic backgrounds, Mailei Zuowu Xuebao
(Journal of Triticeae crops), 26(5): 1-4
Liu P.W., Li Y., He Q.B., and Yang G.S., 2007, Identification of genetic
markers for the pol CMS restorer gene in
Brassica napus
L., Zhongguo
Youliao Zuowu Xuebao (Chinese journal of oil crop science), 29(1):
14-19
Liu Y.H., Yan H.F., Yang W.X., Li X., Li Y.N., Meng Q.F., Zhang L.R., Liu
D.Q., and Zhang T., 2008, Molecular diversity of 23 wheat leaf rust
resistance near-isogenic lines determined by sequence-related amplified
polymorphism, Zhongguo Nongye Kexue (Scientia Agricultura Sinica),
41(5), 1333-1340
Long W.H., Gao J.Q., Pu H.M., Qi C.K., Zhang J.F., and Chen S, 2009,
Molecular fingerprints construction of the new rapeseed variety ningza
No.15, Jiangsu Nongye Xuebao (Jiangsu Journal of Agriculture
Science), 25(6): 1238-1242
Ma S.C., Zhang G.S., and Niu N., 2007, Breeding of near-isogenic lines of
multi-ovary character in wheat and their genetic background evaluation,
He Nongxue Bao (Acta Agriculture Nucleatae Sinica), 21(6): 585-588
Pu H.M., Fu S.Z., Qi C.K., Zhang J.F., Wu Y.M., Gao J.Q., and Chen X.J.,
2001, Inheritance of divided leaf trait of rapeseed (
Brassica napus
) and
application in hybrid breeding, Zhongguo Youliao Zuowu Xuebao
(Chinese journal of oil crop science), 23(3): 60-61
Shen S.Q., Zeng Y.W., and Pu X.Y., 2004, Evaluation of near-isogenic lines
of cold tolerance at booting stage of rice, Xinan Nongye Xuebao
(Southwest China Journal of Agriculture Sciences), 17(S): 18-23
Toshiyuki K., Toshio Y., Naoki T., Masakazu K., and Hideko M., 2003, Fine
genetic mapping of the nuclear gene, Rf-1, that restores the bt-type
cytoplasmic male sterility in rice (Oryza sativa L.) by PCR-based markers,
Euphytica, 129(2): 241-247 http://dx.doi.org/10.1023/A:1021915611210
Von Diethard T., 1989, Hypervariabflity of simple sequences as a general